WASHINGTON In the coming years, the U.S. Navy will be faced with a decision that will radically shape the carrier air wing: Is the service willing to sacrifice dozens of new Super Hornet jets for the promise of a sixth-generation fighter in the 2030s?

The Navy is opting to buy a final 24 F/A-18E/Fs in fiscal 2021, slashing a planned purchase of at least 36 Super Hornets that would have spanned FY22 through FY24. The move will save $4.5 billion, which the service plans to redirect to its sixth-generation fighter program, known as Next Generation Air Dominance, or F/A-XX.

However, the decision may not be as clear cut or final as budget documents make it seem.

The Navy is at the very start of the NGAD development process, having completed an analysis of alternatives in June 2019, as well as broad requirements and guidance for a concept of operations. The effort is now in the concept development phase, during which defense companies explore ideas that balance advanced air dominance capabilities and long-term affordability/sustainment, said Navy spokesman Capt. Danny Hernandez.

But with an economic downturn potentially leading to even more pressure on the defense budget, the Navy may not have the funds to proceed with NGAD as a clean-sheet fighter jet.

Although the Navy would like to start developing the next generation of aircraft ... I just dont think and increasingly people in the department are thinking theres not going to be the money to devote to this next generation of fighter, said Bryan Clark, an analyst with the Hudson Institute and a retired naval officer.

I think they are going to fall back to looking at F/A-XX as a modification or an evolution of the F-35, he said. Instead of the other half of the air wing being some new aircraft, youll have a combination of F-35Cs and then some modified version of the F-35 or a modified Super Hornet."

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Jerry Hendrix, a retired Navy captain and an analyst with the Telemus Group, said the services enthusiasm for F/A-XX is a sign of a continued preference for manned aviation as well as a desire to shut out any hope of fielding a long-range, penetrating strike drone.

Ive always been in favor of extending the Hornet production line because it is solid and stable, Hendrix said. But the extension was based on the proviso that were extending in order to get to an unmanned combat aerial vehicle. If it was an extension to get to the next manned fighter were missing the idea of what the future competitive environment, or really the present competitive environment, is all about.

The Navy has grappled in prior years with the question of whether to cease production of the Super Hornet in favor of a future fighter, and it is an argument that lawmakers are wary of.

The Navy first planned to stop buying the F/A-18 in its FY15 budget a decision made to fund the transition to the F-35. But technical issues and delays pushed out the fielding of the Navys F-35C takeoff-and-landing model for aircraft carriers to 2018, leaving the service dependent on a fleet of aging, battle-worn F/A-18s in need of a service-life extension. The Navy ended up listing the F/A-18 on its unfunded priorities list, and Congress followed by funding enough Super Hornets to keep Boeings line running.

If we go back a few years and we look at what happened when we thought we were going to plant the F-35, we let the F-18 slide down, Rep. Donald Norcross, D-N.J., said in a March 10 hearing. Norcross is the chairman of the House Subcommittee on Tactical Air and Land Forces.

The process of standing up the F-35C was much slower than expected, and the Navy ended up buying additional F/A-18s to bridge the capability gap, he said. Yet, here we are getting ready to curtail 36 Super Hornets because we are expecting, you know, the F/A-XX to come online, he added.

Asked how he could be confident that F/A-XX would stay on schedule, Rear Adm. Gregory Harris, the Navys director of air warfare, said he could provide lawmakers a more detailed defense of the Navys Next Generation Air Dominance family of systems in a classified setting.

Were working closely with the Air Force to ensure the systems that we put on that have the [technology readiness level] that gives us confidence that we can achieve that aircraft on time in the early 2030s to replace the F/A-18E/F as it reaches the end of its service life, Harris said.

Missouri Rep. Vicky Hartzler, the subcommittees top Republican, pointed out that the Navy already has a fighter shortfall of about 49 aircraft, with additional F/A-18s being pulled from the operational fleet into a service-life extension period that will take at least a year.

I feel like this is too much operational risk, she said. If you add all those up, this is a severe shortage that we are experiencing, and if you dont account for the attrition rate, actually in combat we would have a very large gap there potentially.

James Geurts, the Navys assistant secretary for research, development and acquisition, said there is always risk when transitioning from legacy to new aircraft, but that improved mission-capable rates and a steady flow of jets moving through upgrades will help balance the shortfall.

Were taking risk until the late 2020s. I think 2029 is when we will get to the full fighter inventory, and so we had to take some risk as we balance that, he said.

The most likely scenario is that, as the Navy presses forward with its plan to curtail funding for F/A-18s, Congress will simply continue buying more of them, Hendrix said.

But one unanswered question is whether lawmakers will also intervene to force the Navy to consider a wider range of aviation options that could give the carrier air wing longer legs.

Im hopeful that there will be a broader conversation, led by Congress and the administration, perhaps together, to say: We are looking at the future security environment. We are looking at the Chinese threat. We are looking at whats happening in Taiwan, whats happening [in] Hong Kong and within the first island chain, and we really need to have this new capability of long-range, penetrating strike, Hendrix said.

What I do realize is that because the Navy is very conservative right now in how its approaching its procurement programs, the Navy will not be the one to say we need this mission.

No matter what the Navy decides, it could impact its procurement of the MQ-25 unmanned tanker drone currently under development by Boeing. Hendrix sees the MQ-25 program as a likely bill payer, particularly if the service continues to buy Hornets.

What was the reason for the MQ-25? It was to take the strain off from the Hornets, which were being used to refuel other F/A-18s and burning through their service lives faster than anticipated, Hendrix said. When you reopen the Hornet production line and you add 120-something new Hornets, you actually took that strain off the Hornet fleet. So there really isnt a requirement now for a recovery tanker.

Clark agrees that the Navy should develop a long-range unmanned combat aircraft but is unlikely to do so.

But should the Navy choose not to proceed with an F/A-XX program, Clark believes the service could funnel some of that money into modifying the MQ-25 to supplement strike, intelligence, surveillance and reconnaissance capacities, and could even end buy more MQ-25s than planned.

The MQ-25 program, once it gets fielded and proven out, I could see the Navy expanding it, he said. I think the operational and programmatic pressures have driven the Navy to embrace the MQ-25, and because its a complement to the manned aircraft, its generated less resentment among the manned aviation community.

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At a budgetary crossroads, the US Navy's aviation wing must choose between old and new - DefenseNews.com

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Global Impact of COVID-19 Outbreak on Milk Thistle Health Tonic, Market Segmentation by Product:Since the COVID-19 virus outbreak in December 2019, the disease has spread to almost 200 countries around the globe with the World Health Organization declaring it a public health emergency. The global impacts of the coronavirus disease 2019 (COVID-19) are already starting to be felt, and will significantly affect the Milk Thistle Health Tonic market in 2020. COVID-19 can affect the global economy in three main ways: by directly affecting production and demand, by creating supply chain and market disruption, and by its financial impact on firms and financial markets. The outbreak of COVID-19 has brought effects on many aspects, like flight cancellations; travel bans and quarantines; restaurants closed; all indoor events restricted; over forty countries state of emergency declared; massive slowing of the supply chain; stock market volatility; falling business confidence, growing panic among the population, and uncertainty about future. This report also analyses the impact of Coronavirus COVID-19 on the Milk Thistle Health Tonic industry. Prior to COVID-19, the global market for Milk Thistle Health Tonic was anticipated to grow from US$ XX million in 2020 to US$ XX million by 2026; it is expected to grow at a CAGR of xx% during 20212026, whereas post-COVID-19 scenario, the market for Milk Thistle Health Tonic is projected to grow from US$ XX million in 2020 (a change by ~XX% compared to market estimated for 2020 before the outbreak of COVID-19) to US$ XX billion by 2026; it is expected to grow at a CAGR of XX% during 20212026. QY Research has recently curated a research report titled, Global Milk Thistle Health Tonic Market Research Report 2020. The report is structured on primary and secondary research methodologies that derive historic and forecast data. The global Milk Thistle Health Tonic market is growing remarkably fast and is likely to thrive in terms of volume and revenue during the forecast period. Readers can gain insight into the various opportunities and restraints shaping the market. The report demonstrates the progress and bends that will occur during the forecast period. Global Milk Thistle Health Tonic Market: Drivers and Restrains The research report has incorporated the analysis of different factors that augment the markets growth. It constitutes trends, restraints, and drivers that transform the market in either a positive or negative manner. This section also provides the scope of different segments and applications that can potentially influence the market in the future. The detailed information is based on current trends and historic milestones. This section also provides an analysis of the volume of sales about the global market and also about each type from 2015 to 2026. This section mentions the volume of sales by region from 2015 to 2026. Pricing analysis is included in the report according to each type from the year 2015 to 2026, manufacturer from 2015 to 2020, region from 2015 to 2020, and global price from 2015 to 2026. A thorough evaluation of the restrains included in the report portrays the contrast to drivers and gives room for strategic planning. Factors that overshadow the market growth are pivotal as they can be understood to devise different bends for getting hold of the lucrative opportunities that are present in the ever-growing market. Additionally, insights into market experts opinions have been taken to understand the market better. 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This section analyses region-wise revenue and volume for the forecast period of 2015 to 2026. These analyses will help the reader to understand the potential worth of investment in a particular region. Global Milk Thistle Health Tonic Market: Competitive Landscape This section of the report identifies various key manufacturers of the market. It helps the reader understand the strategies and collaborations that players are focusing on combat competition in the market. The comprehensive report provides a significant microscopic look at the market. The reader can identify the footprints of the manufacturers by knowing about the global revenue of manufacturers, the global price of manufacturers, and sales by manufacturers during the forecast period of 2015 to 2019. Following are the segments covered by the report are:, Tablets, Capsules, Others By Application:, Dietary Supplement, Health Food Key Players: The Key manufacturers that are operating in the global Milk Thistle Health Tonic market are:, Health Genesis, Pure Encapsulations, Regis, Solgar, Aksuvital, BEC, NC, Life Extension, Swisse, HerbsofGold Competitive Landscape The analysts have provided a comprehensive analysis of the competitive landscape of the global Milk Thistle Health Tonic market with the company market structure and market share analysis of the top players. The innovative trends and developments, mergers and acquisitions, product portfolio, and new product innovation to provide a dashboard view of the market, ultimately providing the readers accurate measure of the current market developments, business strategies, and key financials.

Global Impact of COVID-19 Outbreak on Milk Thistle Health Tonic, Market Segmentation by Application: Dietary Supplement, Health Food Key Players: The Key manufacturers that are operating in the global Milk Thistle Health Tonic market are:, Health Genesis, Pure Encapsulations, Regis, Solgar, Aksuvital, BEC, NC, Life Extension, Swisse, HerbsofGold Competitive Landscape The analysts have provided a comprehensive analysis of the competitive landscape of the global Milk Thistle Health Tonic market with the company market structure and market share analysis of the top players. The innovative trends and developments, mergers and acquisitions, product portfolio, and new product innovation to provide a dashboard view of the market, ultimately providing the readers accurate measure of the current market developments, business strategies, and key financials.

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Table od Content

1 Milk Thistle Health Tonic Market Overview1.1 Product Overview and Scope of Milk Thistle Health Tonic1.2 Covid-19 Implications on Milk Thistle Health Tonic Segment by Type1.2.1 Global Milk Thistle Health Tonic Sales Growth Rate Comparison by Type (2021-2026)1.2.2 Tablets1.2.3 Capsules1.2.4 Others1.3 Covid-19 Implications on Milk Thistle Health Tonic Segment by Application1.3.1 Milk Thistle Health Tonic Sales Comparison by Application: 2020 VS 20261.3.2 Dietary Supplement1.3.3 Health Food1.4 Covid-19 Implications on Global Milk Thistle Health Tonic Market Size Estimates and Forecasts1.4.1 Global Milk Thistle Health Tonic Revenue 2015-20261.4.2 Global Milk Thistle Health Tonic Sales 2015-20261.4.3 Milk Thistle Health Tonic Market Size by Region: 2020 Versus 20261.5 Coronavirus Disease 2019 (Covid-19): Milk Thistle Health Tonic Industry Impact1.5.1 How the Covid-19 is Affecting the Milk Thistle Health Tonic Industry1.5.1.1 Milk Thistle Health Tonic Business Impact Assessment Covid-191.5.1.2 Supply Chain Challenges1.5.1.3 COVID-19s Impact On Crude Oil and Refined Products1.5.2 Market Trends and Milk Thistle Health Tonic Potential Opportunities in the COVID-19 Landscape1.5.3 Measures / Proposal against Covid-191.5.3.1 Government Measures to Combat Covid-19 Impact1.5.3.2 Proposal for Milk Thistle Health Tonic Players to Combat Covid-19 Impact 2 Covid-19 Implications on Global Milk Thistle Health Tonic Market Competition by Manufacturers2.1 Global Milk Thistle Health Tonic Sales Market Share by Manufacturers (2015-2020)2.2 Global Milk Thistle Health Tonic Revenue Share by Manufacturers (2015-2020)2.3 Global Milk Thistle Health Tonic Average Price by Manufacturers (2015-2020)2.4 Manufacturers Milk Thistle Health Tonic Manufacturing Sites, Area Served, Product Type2.5 Milk Thistle Health Tonic Market Competitive Situation and Trends2.5.1 Milk Thistle Health Tonic Market Concentration Rate2.5.2 Global Top 5 and Top 10 Players Market Share by Revenue2.5.3 Market Share by Company Type (Tier 1, Tier 2 and Tier 3)2.6 Manufacturers Mergers & Acquisitions, Expansion Plans2.7 Primary Interviews with Key Milk Thistle Health Tonic Players (Opinion Leaders) 3 Covid-19 Implications on Milk Thistle Health Tonic Retrospective Market Scenario by Region3.1 Global Milk Thistle Health Tonic Retrospective Market Scenario in Sales by Region: 2015-20203.2 Global Milk Thistle Health Tonic Retrospective Market Scenario in Revenue by Region: 2015-20203.3 North America Milk Thistle Health Tonic Market Facts & Figures by Country3.3.1 North America Milk Thistle Health Tonic Sales by Country3.3.2 North America Milk Thistle Health Tonic Sales by Country3.3.3 U.S.3.3.4 Canada3.4 Europe Milk Thistle Health Tonic Market Facts & Figures by Country3.4.1 Europe Milk Thistle Health Tonic Sales by Country3.4.2 Europe Milk Thistle Health Tonic Sales by Country3.4.3 Germany3.4.4 France3.4.5 U.K.3.4.6 Italy3.4.7 Russia3.5 Asia Pacific Milk Thistle Health Tonic Market Facts & Figures by Region3.5.1 Asia Pacific Milk Thistle Health Tonic Sales by Region3.5.2 Asia Pacific Milk Thistle Health Tonic Sales by Region3.5.3 China3.5.4 Japan3.5.5 South Korea3.5.6 India3.5.7 Australia3.5.8 Taiwan3.5.9 Indonesia3.5.10 Thailand3.5.11 Malaysia3.5.12 Philippines3.5.13 Vietnam3.6 Latin America Milk Thistle Health Tonic Market Facts & Figures by Country3.6.1 Latin America Milk Thistle Health Tonic Sales by Country3.6.2 Latin America Milk Thistle Health Tonic Sales by Country3.6.3 Mexico3.6.3 Brazil3.6.3 Argentina3.7 Middle East and Africa Milk Thistle Health Tonic Market Facts & Figures by Country3.7.1 Middle East and Africa Milk Thistle Health Tonic Sales by Country3.7.2 Middle East and Africa Milk Thistle Health Tonic Sales by Country3.7.3 Turkey3.7.4 Saudi Arabia3.7.5 U.A.E 4 Global Milk Thistle Health Tonic Historic Market Analysis by Type4.1 Global Milk Thistle Health Tonic Sales Market Share by Type (2015-2020)4.2 Global Milk Thistle Health Tonic Revenue Market Share by Type (2015-2020)4.3 Global Milk Thistle Health Tonic Price Market Share by Type (2015-2020)4.4 Global Milk Thistle Health Tonic Market Share by Price Tier (2015-2020): Low-End, Mid-Range and High-End 5 Global Milk Thistle Health Tonic Historic Market Analysis by Application5.1 Global Milk Thistle Health Tonic Sales Market Share by Application (2015-2020)5.2 Global Milk Thistle Health Tonic Revenue Market Share by Application (2015-2020)5.3 Global Milk Thistle Health Tonic Price by Application (2015-2020) 6 Company Profiles and Key Figures in Milk Thistle Health Tonic Business6.1 Health Genesis6.1.1 Corporation Information6.1.2 Health Genesis Description, Business Overview and Total Revenue6.1.3 Health Genesis Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.1.4 Health Genesis Products Offered6.1.5 Health Genesis Recent Development6.2 Pure Encapsulations6.2.1 Pure Encapsulations Corporation Information6.2.2 Pure Encapsulations Description, Business Overview and Total Revenue6.2.3 Pure Encapsulations Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.2.4 Pure Encapsulations Products Offered6.2.5 Pure Encapsulations Recent Development6.3 Regis6.3.1 Regis Corporation Information6.3.2 Regis Description, Business Overview and Total Revenue6.3.3 Regis Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.3.4 Regis Products Offered6.3.5 Regis Recent Development6.4 Solgar6.4.1 Solgar Corporation Information6.4.2 Solgar Description, Business Overview and Total Revenue6.4.3 Solgar Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.4.4 Solgar Products Offered6.4.5 Solgar Recent Development6.5 Aksuvital6.5.1 Aksuvital Corporation Information6.5.2 Aksuvital Description, Business Overview and Total Revenue6.5.3 Aksuvital Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.5.4 Aksuvital Products Offered6.5.5 Aksuvital Recent Development6.6 BEC6.6.1 BEC Corporation Information6.6.2 BEC Description, Business Overview and Total Revenue6.6.3 BEC Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.6.4 BEC Products Offered6.6.5 BEC Recent Development6.7 NC6.6.1 NC Corporation Information6.6.2 NC Description, Business Overview and Total Revenue6.6.3 NC Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.4.4 NC Products Offered6.7.5 NC Recent Development6.8 Life Extension6.8.1 Life Extension Corporation Information6.8.2 Life Extension Description, Business Overview and Total Revenue6.8.3 Life Extension Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.8.4 Life Extension Products Offered6.8.5 Life Extension Recent Development6.9 Swisse6.9.1 Swisse Corporation Information6.9.2 Swisse Description, Business Overview and Total Revenue6.9.3 Swisse Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.9.4 Swisse Products Offered6.9.5 Swisse Recent Development6.10 HerbsofGold6.10.1 HerbsofGold Corporation Information6.10.2 HerbsofGold Description, Business Overview and Total Revenue6.10.3 HerbsofGold Milk Thistle Health Tonic Sales, Revenue and Gross Margin (2015-2020)6.10.4 HerbsofGold Products Offered6.10.5 HerbsofGold Recent Development 7 Milk Thistle Health Tonic Manufacturing Cost Analysis7.1 Milk Thistle Health Tonic Key Raw Materials Analysis7.1.1 Key Raw Materials7.1.2 Key Raw Materials Price Trend7.1.3 Key Suppliers of Raw Materials7.2 Proportion of Manufacturing Cost Structure7.3 Manufacturing Process Analysis of Milk Thistle Health Tonic7.4 Milk Thistle Health Tonic Industrial Chain Analysis 8 Marketing Channel, Distributors and Customers8.1 Marketing Channel8.2 Milk Thistle Health Tonic Distributors List8.3 Milk Thistle Health Tonic Customers 9 Market Dynamics 9.1 Market Trends 9.2 Opportunities and Drivers 9.3 Challenges 9.4 Porters Five Forces Analysis 10 Global Market Forecast10.1 Global Milk Thistle Health Tonic Market Estimates and Projections by Type10.1.1 Global Forecasted Sales of Milk Thistle Health Tonic by Type (2021-2026)10.1.2 Global Forecasted Revenue of Milk Thistle Health Tonic by Type (2021-2026)10.2 Milk Thistle Health Tonic Market Estimates and Projections by Application10.2.1 Global Forecasted Sales of Milk Thistle Health Tonic by Application (2021-2026)10.2.2 Global Forecasted Revenue of Milk Thistle Health Tonic by Application (2021-2026)10.3 Milk Thistle Health Tonic Market Estimates and Projections by Region10.3.1 Global Forecasted Sales of Milk Thistle Health Tonic by Region (2021-2026)10.3.2 Global Forecasted Revenue of Milk Thistle Health Tonic by Region (2021-2026)10.4 North America Milk Thistle Health Tonic Estimates and Projections (2021-2026)10.5 Europe Milk Thistle Health Tonic Estimates and Projections (2021-2026)10.6 Asia Pacific Milk Thistle Health Tonic Estimates and Projections (2021-2026)10.7 Latin America Milk Thistle Health Tonic Estimates and Projections (2021-2026)10.8 Middle East and Africa Milk Thistle Health Tonic Estimates and Projections (2021-2026) 11 Research Finding and Conclusion 12 Methodology and Data Source 12.1 Methodology/Research Approach 12.1.1 Research Programs/Design 12.1.2 Market Size Estimation 12.1.3 Market Breakdown and Data Triangulation 12.2 Data Source 12.2.1 Secondary Sources 12.2.2 Primary Sources 12.3 Author List 12.4 Disclaimer

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Impact of COVID-19 Outbreak on Milk Thistle Health Tonic, Market Size, Share, Growth, Trends and Forecast 2026 by Top Manufacturers, Instruments...

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My summer countdown usually starts on the first day of fall. However, with social distancing still in place and travel completely off the cards for the foreseeable future, it's tough to get excited about what's arguably the best time of year.

Throughout quarantine, beauty products have given me a little bit of comfort and madethe stressand challenges of our current reality seem more manageable.

But even though everyone'sdaily routines have changed and the beauty industry has been greatly impacted by COVID-19, brands haven't stopped launching new products.

RELATED:Shopping for Makeup Post COVID-19 Lockdown Will Never Be the Same

This month's just-launched and soon-to-launch makeup, skincare, and haircare products include a number of treatments that are perfect for taking a time out and indulging in a little TLC in isolation. Briogeo's repairing hair mask, HoliFrog's glow-boosting cleanser, and Gucci Westman'svelvety eyeshadows are just a few examples.

Ahead, 15 new beauty products to give yourself some extra self-care while stuck at home.

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The 15 Best New Products to Try in Isolation This Month - InStyle

Recommendation and review posted by Bethany Smith

Researchers are learning more about genetic aberrations common in the rare but clinically aggressive hematological cancer blastic plasmacytoid dendritic cell neoplasm. There is one targeted therapy approved by the U.S. Food and Drug Administration: Elzonris (tagraxofusp-erzs, Stemline). However, more treatment options are needed to improve the cancers clinical outcome, according to a review published May 2020 in Critical Reviews Oncology/Hematology.1

Dermatologists might be the first providers to encounter patients with blastic plasmacytoid dendritic cell neoplasm because more than 70% of these patients have cutaneous lesions. Those lesions often are asymptomatic and vary in size. The skin lesions tend to have nodules, plaques or bruise-like areas, a brown to violet color and might be solitary or multifocal, according to the authors.

Blastic plasmacytoid dendritic cell neoplasm often originates from type 2 myeloid-derived resting plasmacytoid dendritic cell precursors. Recent research suggests providers can diagnose the cancer when patients express at least four of five plasmacytoid dendritic cell specific markers, CD4, CD56, CD123, TCL1 and BDCA-2, without expressing myeloid, T-cell or B-cell lineage markers.

Commonly, [blastic plasmacytoid dendritic cell neoplasm] is characterized by high CD123 expression, aberrant NF-B [nuclear factor-B] activation, dependence on TCF4-/BRD4-network, and deregulated cholesterol metabolism, they wrote.

Despite advancing knowledge about the cancer type, patients median overall survival remains at 12 to 14 months, according to the paper. Conventional treatment approaches include chemotherapy, radiotherapy and ultimately hematopoietic stem cell transplantation. The challenges with conventional therapies are while blastic plasmacytoid dendritic cell neoplasm is sensitive to some chemotherapy regimens, patient relapse is high at more than 60%. And many patients with blastic plasmacytoid dendritic cell neoplasm are too old or frail to have intensive chemotherapy or hematopoietic stem cell transplantation, according to the authors.

Recently, the most attractive agent for [blastic plasmacytoid dendritic cell neoplasm] is tagraxofusp, which is composed of the catalytic and translocation domains of diphtheria toxin (DT) fused to interleukin-3 (IL-3), the authors wrote.

Blastic plasmacytoid dendritic cell neoplasm cells overexpress interleukin-3 receptor subunit alpha (IL3RA, also called CD123). Elzonris, or tagraxofusp-erzs, is a CD123-directed cytotoxin given intravenously, which is used to treat blastic plasmacytoid dendritic cell neoplasm in adults and in pediatric patients 2 years and older.

Researchers reported in a study of 47 blastic plasmacytoid dendritic cell neoplasm patients published in 2019 in the New England Journal of Medicine that tagraxofusp led to clinical responses in untreated and relapsed patients.2 The overall response rate with tagraxofusp was 90% and the primary outcome of complete response and clinical complete response was 72% among the previously untreated patients. Overall response was 67% in the previously treated patients. Serious adverse events including capillary leak syndrome, hepatic dysfunction and thrombocytopenia were common, according to the NEJM paper.

More targeted therapies are needed to treat blastic plasmacytoid dendritic cell neoplasm, but many potential therapeutic agents are not advancing to clinical trials, according to authors of the paper in Critical Reviews Oncology/Hematology.

Common blastic plasmacytoid dendritic cell neoplasm characteristics are genetically heterogeneous and provide valuable drug targets, according to the authors.

Apart from aberrant activation of NF-B signaling pathway, which is highly dependent on TCF4- and BRD4- transcriptional networks, cholesterol metabolism deregulation and CD123 expression, defects of DNA damage repair and mitosis are new, potential common features of the cancer. Corresponding therapies might be promising, the authors wrote.

Venetoclax, anti-CD123 CAR-T, XmAb14045 and IMGN632 are in clinical trials for blastic plasmacytoid dendritic cell neoplasm. But the authors noted that bortezomib, lenalidomide, 5-aza and pralatrexate could easily be pushed to the front line of the cancers treatment.

Disclosures:

The authors report no relevant disclosures.

References:

1. Zhang X, Sun J, Yang M, Wang L, Jin J. New perspectives in genetics and targeted therapy for blastic plasmacytoid dendritic cell neoplasm. Crit Rev Oncol Hematol. 2020 May;149:102928.2. Pemmaraju N, Lane AA, Sweet KL, et al. Tagraxofusp in Blastic Plasmacytoid Dendritic-Cell Neoplasm. N Engl J Med. 2019;380(17):1628-1637.

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Genetic features pave way for targeted BPDCN therapies - Dermatology Times

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DetailsCategory: DNA RNA and CellsPublished on Monday, 01 June 2020 18:32Hits: 165

The Phase 2 study will assess the safety and efficacy of Longeveron's stem cell treatment under Japan's accelerated regulatory pathway for regenerative medicine.

MIAMI, FL, USA I June 1, 2020 I Longeveron LLC announced today that Japan's Pharmaceutical and Medical Devices Agency (PMDA) (the Japanese agency akin to the United States' Food & Drug Administration) approved a Clinical Trial Notification (CTN) application (akin to an Investigational New Drug Application or "IND" in the US regulatory system), approving the initiation of a Phase 2 clinical trial evaluating the safety and efficacy of Longeveron's Mesenchymal Stem Cells (LMSCs) for the treatment of Aging Frailty in Japanese patients. This is another key milestone for Longeveron's Aging Frailty program, which includes two ongoing Phase 2 clinical trials in the U.S.

"We are extremely pleased to achieve this significant milestone," said Geoff Green, President of Longeveron."This study is designed to determine whether the transplant of donor-derived mesenchymal stem cells can improve healthspan in mild to moderately frail patients, thereby improving functionality and potentially lowering their risk of disability, and dependence on others for care."

Aging Frailty is a common, but reversible, life-threatening geriatric condition affecting millions of Japanese over the age of 65.Frail individuals are vulnerable to adverse health outcomes compared to their age-matched peers despite sharing similar comorbidities and demographics.Clinically, frailty manifests as a combination of symptoms that may include loss of muscle and decreased strength, slowed walking (sarcopenia), lower activity and energy levels, poor endurance, nutritional deficiencies, weight loss and fatigue.Collectively, these lead to overall decline in functionality, and increased risk of disability, dependency, and death.

"The biology of frailty is complex, and includes diminished stem cell activity, reduced ability to repair and regenerate tissue, and immunosenescence (deterioration of the immune system) and chronic systemic inflammation," said Dr. Anthony Oliva, Senior Scientist at Longeveron. "LMSCs have multiple mechanisms of action that can potentially address all of these issues, and thus make them extremely attractive as a therapeutic candidate for the unmet medical need of Aging Frailty."

The planned study is an investigator-initiated, randomized, double-blind, placebo-controlled design,and will be conducted at Juntendo University Hospital (Tokyo) and Japan's National Center for Geriatrics and Gerontology (NCGG) in Nagoya.The study's Principal Investigator, Dr. Hidenori Arai, President of the NCGG, commented that "Japan has one of the oldest and fastest aging societies in the world, with nearly 30% of Japan's citizens over the age of 65.Preventing and reversing functional decline associated with frailty is one of the nation's top priorities, and Longeveron's regenerative medicine approach is an exciting and innovative potential therapeutic option.With the disproportionate infection and mortality rate of older people with COVID-19 and Influenza infection, it is critically important to rapidly test treatments that may be effective."

In Japan, the "Pharmaceutical and Medical Device Act" and the "Act on the Safety of Regenerative Medicine" came into effect in 2014. Under this system, a "Time-limited Conditional Approval" option exists, which allows a manufacturer to conditionally sell regenerative medicine products while proceeding with its Phase 3 clinical trial.

Longeveron's Aging Frailty Research Program

Longeveron sponsors the most extensive and advanced Aging Frailty clinical research program in the world, with more than 200 patients treated with LMSCs worldwide.In the U.S., two clinical trials are currently ongoing:

About LMSCs

Longeveron Allogeneic Mesenchymal Stem Cells (LMSCs) is a regenerative medicine product sourced from the bone marrow of young healthy adult donors.LMSCs are culture expanded under the FDA's current good manufacturing practices (cGMP) to high standards, and maintained as individual "off-the-shelf" doses.

About Longeveron LLC

Longeveron (www.longeveron.com) is a regenerative medicine therapy company founded in 2014. Longeveron's mission is to provide biological solutions for aging-related diseases and life-threatening conditions, and is dedicated to developing safe and effective cell-based therapeutics for unmet medical needs such as Aging Frailty, the Metabolic Syndrome, Alzheimer's Disease, Acute Respiratory Distress Syndrome (ARDS) from COVID-19 infection, and congenital heart defects in children (hypoplastic left heart syndrome).

SOURCE: Longeveron

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Longeveron Announces Japanese Approval of Clinical Trial for Treatment of Aging Frailty With Longeveron's Stem Cells | DNA RNA and Cells | News...

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The art (and existing science) of regenerative medicine in equine practice, and whats to come

Regenerative therapy is an umbrellaterm encompassing any method that encourages the body to self- heal. Because it is drawing onits own properties, healing tissue more closely resembles native tissue than weak, disorganized scar tissue typically seen post-injury.

The goal is to allow restoration of normal function and structure of the injured tissue to allow horses to perform at their previous level, whatever that might be, with a reduced risk of reinjury, says Kyla Ortved, DVM, PhD, Dipl. ACVS, ACVSMR, assistant professor of large animal surgery at the University of Pennsylvanias New Bolton Center, in Kennett Square.

She says the three main components of regenerative medicine that help tissues self-heal include:

A specific therapy may incorporate some or all three of these components, says Ortved.

Due to the regenerative therapy industrys popularity and continued growth, many articles weve published review recent laboratory studies about stem cell production and data on efficacy andsafety (you can find them at TheHorse.com/topics/regenerative-medicine). Here, well review the basics of three regenerative modalities commonly used in equine medicine and when veterinarians and horse owners might consider each.

With this approach the practitioner collects blood from a horse and processes it using a commercial system that concentrates the platelets. When he or she injects that concentrated platelet product back into the horse, granules within the platelets release an array of growth factors that aim to facilitate and modulate the healing process. Specifically, granule-derived growth factors encourage target tissue cells at the injury site to migrate and proliferate, improve extracellular matrix synthesis, and stimulate blood vessel development.

Recently, leukocyte-reduced PRP hasbecome many equine veterinarians PRP product of choice. These preparations contain fewer white blood cells (leukocytes) and, reportedly, inflammatory mediators than normal PRP products do. These mediators break tissues down, effectively counteracting the anabolic (tissue-building) effects of the platelets and their granules.

Veterinarians can easily prepare ACS by collecting a blood sample from the patient, then incubating it with special commercially available glass beads to stimulate interleukin-1 receptor antago- nist protein (IRAP) production. Theythen inject the resultant IRAP-rich serumsample back into the patient at the target location or injury site. This protein blocks the action of interleukin-1, a powerful and damaging pro-inflammatory mediator. Additionally, glass bead incubation stimulates the production of anti-inflammatory mediators and growth factors similar to those found in PRP.

Ortved says its important to remember that all biologics, including PRP and IRAP, contain various concentrations of growth factors and bioactive protein.

Remember, they are made from your horses blood and, therefore, containall of the components in blood, just in varying concentrations, she says.

Regenerative therapies that contain highconcentrations of IRAP include IRAP II, autologous protein solution (APS), and bone marrow aspirate concentrate (BMAC).

In certain tissues, such as adipose (fat) and bone marrow, we can find specific cells that have the ability to self-renew and grow more than 200 types of body cells. Veterinarians can isolate these cells, called stem cells or progenitor cells, and either:

Perhaps more important than theirability to differentiate into other celltypes, stem cells have powerful anti-inflammatoryproperties and play acentral role in coordinating healing in alltypes of tissues through cell-to-cell signaling,Ortved says.

Which of these three modality typeswill provide the most benefit to yourhorse depends on a variety of factors thatyou and your veterinarian will consider.

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Regenerative Therapies: Helping Horses Self-Heal The Horse - TheHorse.com

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Stem Cell Assay Market: Snapshot

Stem cell assay refers to the procedure of measuring the potency of antineoplastic drugs, on the basis of their capability of retarding the growth of human tumor cells. The assay consists of qualitative or quantitative analysis or testing of affected tissues andtumors, wherein their toxicity, impurity, and other aspects are studied.

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With the growing number of successfulstem cell therapytreatment cases, the global market for stem cell assays will gain substantial momentum. A number of research and development projects are lending a hand to the growth of the market. For instance, the University of Washingtons Institute for Stem Cell and Regenerative Medicine (ISCRM) has attempted to manipulate stem cells to heal eye, kidney, and heart injuries. A number of diseases such as Alzheimers, spinal cord injury, Parkinsons, diabetes, stroke, retinal disease, cancer, rheumatoid arthritis, and neurological diseases can be successfully treated via stem cell therapy. Therefore, stem cell assays will exhibit growing demand.

Another key development in the stem cell assay market is the development of innovative stem cell therapies. In April 2017, for instance, the first participant in an innovative clinical trial at the University of Wisconsin School of Medicine and Public Health was successfully treated with stem cell therapy. CardiAMP, the investigational therapy, has been designed to direct a large dose of the patients own bone-marrow cells to the point of cardiac injury, stimulating the natural healing response of the body.

Newer areas of application in medicine are being explored constantly. Consequently, stem cell assays are likely to play a key role in the formulation of treatments of a number of diseases.

Global Stem Cell Assay Market: Overview

The increasing investment in research and development of novel therapeutics owing to the rising incidence of chronic diseases has led to immense growth in the global stem cell assay market. In the next couple of years, the market is expected to spawn into a multi-billion dollar industry as healthcare sector and governments around the world increase their research spending.

The report analyzes the prevalent opportunities for the markets growth and those that companies should capitalize in the near future to strengthen their position in the market. It presents insights into the growth drivers and lists down the major restraints. Additionally, the report gauges the effect of Porters five forces on the overall stem cell assay market.

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Global Stem Cell Assay Market: Key Market Segments

For the purpose of the study, the report segments the global stem cell assay market based on various parameters. For instance, in terms of assay type, the market can be segmented into isolation and purification, viability, cell identification, differentiation, proliferation, apoptosis, and function. By kit, the market can be bifurcated into human embryonic stem cell kits and adult stem cell kits. Based on instruments, flow cytometer, cell imaging systems, automated cell counter, and micro electrode arrays could be the key market segments.

In terms of application, the market can be segmented into drug discovery and development, clinical research, and regenerative medicine and therapy. The growth witnessed across the aforementioned application segments will be influenced by the increasing incidence of chronic ailments which will translate into the rising demand for regenerative medicines. Finally, based on end users, research institutes and industry research constitute the key market segments.

The report includes a detailed assessment of the various factors influencing the markets expansion across its key segments. The ones holding the most lucrative prospects are analyzed, and the factors restraining its trajectory across key segments are also discussed at length.

Global Stem Cell Assay Market: Regional Analysis

Regionally, the market is expected to witness heightened demand in the developed countries across Europe and North America. The increasing incidence of chronic ailments and the subsequently expanding patient population are the chief drivers of the stem cell assay market in North America. Besides this, the market is also expected to witness lucrative opportunities in Asia Pacific and Rest of the World.

Global Stem Cell Assay Market: Vendor Landscape

A major inclusion in the report is the detailed assessment of the markets vendor landscape. For the purpose of the study the report therefore profiles some of the leading players having influence on the overall market dynamics. It also conducts SWOT analysis to study the strengths and weaknesses of the companies profiled and identify threats and opportunities that these enterprises are forecast to witness over the course of the reports forecast period.

Some of the most prominent enterprises operating in the global stem cell assay market are Bio-Rad Laboratories, Inc (U.S.), Thermo Fisher Scientific Inc. (U.S.), GE Healthcare (U.K.), Hemogenix Inc. (U.S.), Promega Corporation (U.S.), Bio-Techne Corporation (U.S.), Merck KGaA (Germany), STEMCELL Technologies Inc. (CA), Cell Biolabs, Inc. (U.S.), and Cellular Dynamics International, Inc. (U.S.).

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Stem Cell Assay Market to Witness Growth Acceleration During 2017-2025 - Cole of Duty

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SOUTH SAN FRANCISCO--(BUSINESS WIRE)--Imago BioSciences, Inc. (Imago), a clinical stage biopharmaceutical company developing innovative treatments for myeloid diseases, today announced that positive Phase 2 data from its lead pipeline program bomedemstat (IMG-7289), will be presented at the Virtual Edition of the 25th EHA Annual Congress beginning June 12, 2020.

Title: A PHASE 2 STUDY OF BOMEDEMSTAT (IMG-7289), A LYSINE-SPECIFIC DEMETHYLASE-1 (LSD1) INHIBITOR, FOR THE TREATMENT OF LATER-STAGE MYELOFIBROSIS (MF)

Session Topic: 16. Myeloproliferative Neoplasms

Final Abstract Code: EP1080

The data demonstrates the potential of bomedemstat as a monotherapy in intermediate-2 and high-risk patients with myelofibrosis who have become intolerant of, or resistant to, or are ineligible for a Janus Kinase (JAK) inhibitor.

Imago is currently conducting a Phase 2 study of bomedemstat in five countries. Clinical endpoints include spleen volume reduction, reduction in total symptom scores, and improvement in circulating inflammatory cytokines, anemia, bone marrow fibrosis and blast count. For additional information, visit cliniciatrials.gov (NCT03136185).

About Bomedemstat (IMG-7289)

Bomedemstat is being evaluated in an open-label Phase 2 clinical trial for the treatment of advanced myelofibrosis (MF), a bone marrow cancer that interferes with the production of blood cells. The endpoints include spleen volume reduction and symptom improvement at 12 and 24 weeks of treatment. Bomedemstat is used as monotherapy in patients who are resistant to, intolerant of, or ineligible for a Janus Kinase (JAK) inhibitor.

Bomedemstat is a small molecule developed by Imago BioSciences that inhibits lysine-specific demethylase 1 (LSD1 or KDM1A), an enzyme shown to be vital in cancer stem/progenitor cells, particularly neoplastic bone marrow cells. In non-clinical studies, IMG-7289 demonstrated robust in vivo anti-tumor efficacy across a range of myeloid malignancies as a single agent and in combination with other chemotherapeutic agents. Bomedemstat (IMG-7289) is an investigational agent currently being evaluated in ongoing clinical trials (ClinicalTrials.gov Identifier: NCT03136185 and NCT02842827). Bomedemstat has FDA Orphan Drug and Fast Track Designation for the treatment of myelofibrosis and essential thrombocythemia, and Orphan Drug Designation for treatment of acute myeloid leukemia.

About Imago BioSciences

Imago BioSciences is a clinical-stage biopharmaceutical company focused on discovering and developing novel anti-cancer therapeutics targeting epigenetic enzymes. Imago has developed a series of compounds that inhibit LSD1, an epigenetic enzyme critical for cancer stem cell function and differentiation. Imago is advancing the clinical development of its first LSD1 inhibitor, bomedemstat, for the treatment of myeloid neoplasms including myelofibrosis and essential thrombocythemia. Imago BioSciences is backed by leading strategic and venture investors including a fund managed by Blackstone Life Sciences, Frazier Healthcare Partners, Omega Funds, Amgen Ventures, MRL Ventures Fund, HighLight Capital, Pharmaron, Greenspring Associates and Xeraya Capital. The company is based in South San Francisco, California. To learn more, visit http://www.imagobio.com.

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Imago BioSciences To Present Update on Phase 2 results of Bomedemstat (IMG-7289), a Lysine Specific Demethylase-1 (LSD1) Inhibitor for the Treatment...

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INTRODUCTION

Idiopathic pulmonary fibrosis (IPF) is a rapidly progressive and fatal interstitial pulmonary disease with a dismal median survival time of just 3 years after diagnosis (1, 2). To date, the IPF therapies depend on blocking myofibroblast activation to inhibit collagen I deposition (3, 4). However, the clinical data showed that these therapies remained far from achieving IPF revision. The main reason is that the IPF therapeutics lack an effectively targeted carrier or ignore some of the other risk factors such as the instability and tolerability of type II alveolar epithelial cell (AEC II) (5, 6). The AEC II, which is considered as injured AEC II (7, 8) in the IPF tissues, releases excessive amounts of reactive oxygen species (ROS) that initiate an antifibrinolytic coagulation cascade and promote the overexpression of connective tissue growth factor (CTGF) to provoke myofibroblast overactivation and extracellular matrix (ECM) development and then destroy the lung architecture (911). This situation has inspired us to propose that the combination of modulating superoxide in injured AEC II and antimyofibroblast activation as weeding and uprooting strategy will be a potential therapeutic strategy for synergistic antifibrosis. Furthermore, another limitation is that current therapies are rarely distributed in the lungs, which cannot achieve full therapeutic effect for treating IPF (12). Thus, the development of an effective lung-targeting drug delivery carrier is highly desirable for IPF therapy.

Recently, local preferred therapeutic agents generated using endogenous cells have served as a strong and promising delivery platform for targeting in situ, achieving considerable progress in several diseases (1317). In the inflammatory phase of IPF, precursor circulating monocytes (PCMs) have been found to undergo notable proliferation (18). PCMs and injured AEC II release chemotactic factors that specifically recruit chemokine receptorpositive (CR+) cells including monocyte-derived multipotent cell (MOMC) and guide the MOMC to migrate to injured lung tissues through specific binding to chemokine receptors on cell membrane (19, 20). Furthermore, in addition to this migration characteristic, these MOMCs, which originate from hematopoietic stem cells in the bone marrow, still have multipotency to differentiate into a variety of functional cells, including AEC II and endothelial cell (21, 22), which demonstrates that MOMC has the potential to participate in reestablishing lung functions (23). In addition, chronic hypoxic exposure induces the recruitment of MOMC to the pulmonary circulation, and the cell contributes to improving lung functions by producing angiogenic factors (24). It has been reported that monocytes from patients with IPF also show preconditioned prorepair features (25). In general, MOMC as a precise lung-targeting delivery platform will exhibit encouraging therapeutic effects, leading to the repair or regeneration of injured AEC II for IPF treatment.

In this study, we constructed the programmed therapeutics composed of surface-engineered nanoparticles (PER NPs) loading dual drugs adhered to MOMC (named MOMC/PER) to solve the issues in IPF therapy by improving drug accumulation in injured lung sites and completely destroying the fibrotic signaling network in IPF (Fig. 1). The MOMC/PER delivery platform realized efficacy through programmed modules, which consisted of a homing moiety, responsive release moiety, and retargeting moiety. (i) The homing moiety is the native ability of MOMC/PER to migrate to injured lungs due to the homing characteristic of MOMC. (ii) The responsive release moiety of MOMC/PER is activated by matrix metalloproteinase-2 (MMP-2) overexpression in IPF tissues, resulting in pathology-responsive release of PER NPs with exposed cyclic RGDfc (Arg-Gly-Asp) [c(RGDfc)] from the MOMC. (iii) The retargeting moiety is that exposed c(RGDfc) on PER NPs can anchor to injured AEC II via an interaction between v6 and c(RGDfc) (26), allowing the cytoplasm of the injured AEC II internalize PER NPs. Subsequently, astaxanthin (AST) and trametinib (TRA) are released from PER NPs to achieve a weeding and uprooting therapeutic effect. In general, the sustained injury of epithelial cells and highly heterogeneous myofibroblasts is considered as the most critical variable in achieving complete IPF reversion (27). To validate the above hypothesis, in this study, AST was chosen as an antioxidant by neutralizing superoxide to repair injured AEC II (28), and TRA suppressed the activation of myofibroblast by inhibiting CTGF production for IPF therapy (29). MOMC also participates in treating IPF by repairing injured AEC II to promote regeneration of IPF lungs (21). Overall, MOMC/PER, which mimics the features of chimeric antigen receptor T cell immunotherapy, is a precise lung-targeting platform to reverse IPF by improving drug accumulation due to the outstanding homing ability of MOMC to injured lung sites, and the destruction of the fibrotic signaling network by inhibiting the activation of myofibroblast and repairing injured AEC II to promote the damaged lungs regeneration.

(A) Bioconjugated MOMC/PER was prepared by incubating PER NPs with MOMC. (B) MOMC/PER has multifunctional moieties including a homing moiety, responsive release moiety, and retargeting moiety to reverse IPF. Then, a weeding and uprooting strategy contributes to IPF reversion. (C) Schematic illustration of MOMC/PER for improved drugs accumulation and antifibrotic effect in IPF lung microenvironment.

The quantities of MOMC in serum and lung tissues were significantly increased in IPF mice compared with normal mice (Fig. 2A). The proliferation of MOMC was positively related to IPF progression, which might be because increasing numbers of MOMC would be recruited from the bone marrow to the lesion sites when IPF occurred (30). Motivated by the fact that MOMC has a homing ability, we considered MOMC to be a potential delivery carrier to improve delivery efficiency in IPF treatment under pathological conditions.

(A) The proliferation of Nanog+ cells in serum and lung tissues by ELISA assay. (B) The MOMC phenotypes. The level of TGF- (C) and hydroxyproline (D) in IPF lung tissues, respectively. (E) The level of TGF-/Smad in vitro. (F) Schematic showing the preparation of MOMC/PER. (G) Schematic showing the adhesion of PER NPs to MOMC. (H) SEM images of MOMC and MOMC/PER-DiI. (I) Fluorescent signals of MOMC and PER-DiI NPs by CLSM. (J) The adhesion between MOMC and PER-DiI NPs by flow cytometry. (K) In vitro migration model. The migration ability of MOMC and MOMC/PER in CXCL 12 (L) and CCL 19 (M), respectively. (N) Schematic showing sensitive release of MOMC/PER-DiI triggered by MMP-2. (O) Characterizations by TEM. MOMC is the triangle, and PER-DiI NPs are the arrows. (P) Fluorescent images of MOMC and released PER-DiI NPs by CLSM. (Q) The flow cytometry showed responsive release. (R) Schematic showing the retarget ability of released PER NPs. (S) Characterization of retargeting ability by TEM. (T) The fluorescent images by CLSM. (U) Cellular uptake in A549 by flow cytometry (n = 3). Statistical significance was calculated via one-way analysis of variance (ANOVA).

We first isolated MOMC from the peripheral blood of C57BL/6J male mice of IPF. The morphologies of the MOMC were fusiform (fig. S1). To identify the phenotypes of MOMC isolated from IPF mice, we first investigated the presence of specific markers for MOMC by immunofluorescence staining. The results showed that MOMC expressed CD11b and smooth muscle actin (-SMA) (Fig. 2B), which was consistent with the literature (24). In addition, the MOMC also expressed the stem cell markers CD14 and Nanog protein and the injured AEC IIs marker pro-surfactant protein C (SPC), as shown in Fig. 2B. These results indicated that MOMC was pluripotent cells with stem cell and epithelial celllike properties. It has been reported that MOMC was recruited to damaged lung areas and participated in recovering injured lung normalization through growth factor release to repair injured AEC II (30). In addition, to inspect the potential risk of injecting MOMC into mice, we further investigated the feasibility of using isolated MOMC as a delivery carrier, including measuring the levels of transforming growth factor (TGF-) and hydroxyproline, which are closely related to the development of IPF in vivo. The results displayed approximately onefold reduction in TGF- and hydroxyproline levels in IPF mice treated with MOMC compared with untreated bleomycin (BLM)induced mice, and these indexes were barely changed in normal mice, indicating that MOMC would not induce the occurrence of IPF and partly relieved established IPF (Fig. 2, C and D).

We next prepared PER NPs that contained two target peptides named peptide E5 and c(RGDfc). The poly(lactide-co-glycolide)-block-poly(ethylene glycol) methyl ether maleimide (PLGA-PEG-Mal) and PLGA-PEG-c(RGDfc) (mass ratio, 10:1) were self-assembled by noncovalent interactions of amphiphilic PLGA-PEG copolymer into nanoparticles (31), and then, peptide E5 was bound on the NPs by the Michael reaction (fig. S2A). As determined by 1H nuclear magnetic resonance spectroscopy (fig. S2B) and SDSpolyacrylamide gel electrophoresis (SDS-PAGE) (fig. S2C), we successfully prepared PER NPs, and the grafting rate of peptide E5 in the PER NPs was 43.7%. The PER NPs showed particle sizes of approximately 110 10.39 nm and the zeta potential of 23.37 mV (fig. S2, D and E). In addition, AST and TRA were encapsulated into PER NPs (fig. S2F). The drug loading content of the PER NPs was 1.98 weight % (wt %) for AST and 2.83 wt % for TRA. The sustained release of the loaded AST was 49.5 wt %, and the pH-dependent release of the loaded TRA (weak alkalinity) was 79.6 wt %, which were obtained at pH 5.0 within 72 hours (fig. S2G). Then, we investigated the capacity of MOMC to differentiate into myofibroblast after treatment with PER NPs in vitro. As shown in Fig. 2E, the expression of TGF-/small mother against decapentaplegic (TGF-/Smad), which is molecule in the crucial pathway for myofibroblast activation, was decreased, suggesting that the PER NPs could inhibit MOMC differentiation. The possible reason for the inhibition was that the PER NPs partly covered the TGF- receptor on the MOMC and reduced exogenous TGF- stimulation within 8 hours, and then, the PER NPs could be gradually internalized. The released drugs could reduce TGF- expression of MOMC after 8 hours (fig. S3, A and B).

We next constructed MOMC/PER as a delivery platform/therapeutic carrier (Fig. 2F), and PER NPs loaded with both drugs could specifically adhere to MOMC through the interaction between peptide E5 of the PER NPs and the CXCR4 on the MOMC by a temperature-dependent manner (32, 33). The formation of MOMC/PER was positively correlated with the incubation time within 2 hours (fig. S3, A and B). Moreover, the PER NPs could specifically stick to the surface of the MOMC without internalization by the MOMC within 8 hours (Fig. 2G). The reasons may be that peptide E5 conjugated on the surface of the PER NPs is a long-chain peptide that limits internalization into the MOMC and that CXCR4 is not an endocytic receptor (34). The MOMC/PER had a loading capacity of 4.75 g of TRA and 1.5 g of AST/1 105 cells (fig. S3, C and D). In addition, MOMC cell viability was above 80% with different concentrations of PER NPs and different incubation times (fig. S4, A and B).

To investigate the adhesion of MOMC and PER NPs, the 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine perchlorate (DiI) was loaded into blank PER NPs (PER-DiI NPs) to evaluate adhesion behavior. After incubating with MOMC and PER-DiI NPs for 2 hours, the morphologies of the MOMC/PER-DiI were confirmed by scanning electron microscopy (SEM) (Fig. 2H) and confocal laser scanning microscopy (CLSM) (Fig. 2I). Flow cytometry detection also indicated the formation of MOMC/PER-DiI in that the MOMC labeling green and PER-DiI NPs were collected in the double-positive quadrant (Fig. 2J).

Sequentially, migration via the interaction between a receptor and ligand is the vital characteristic that needs to be retained by MOMC/PER to realize efficient delivery (Fig. 2K). The migratory capability of MOMC/PER was detected by a Transwell invasion assay. The results indicated that the migratory ability of the MOMC/PER was unaffected by PER NPs adhering to the surface of MOMC (Fig. 2, L and M) at all studied concentrations (fig. S4, C and D).

To establish the retargeting ability of PER NPs, MMP-2 overexpressed in IPF tissues was used as an activating trigger to release PER NPs from MOMC/PER. As depicted in Fig. 2N, the separation of PER-DiI NPs from MOMC was well evidenced by transmission electron microscopy (TEM) and CLSM (Fig. 2, O and P) and flow cytometry (Fig. 2Q). We also detected the phenomenon by SDS-PAGE and particle sizes changes (fig. S4, E and F). After PER NPs were released from MOMC/PER, the exposed peptide c(RGDfc) of the PR NPs could retarget v6, which is overexpressed on the surface of injured AEC II (Fig. 2R) (35). Then, we investigated the capacity of injured AEC II to uptake PLGA-PEG-c(RGDfc)coumarin 6 (PR-C6) compared with free C6 and PLGA-PEGC6 (PP-C6) by TEM and CLSM. The internalization of PR-C6 was better than other forms (C6 and PP-C6) (Fig. 2, S to U). In addition, PR-C6 also underwent lysosomal escape (fig. S4G).

The homing ability of MOMC/PER was investigated in IPF models in vivo (Fig. 3A). We first examined the lung accumulation of 1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbocyanine iodide (DiR)loaded into blank PER NPs (PER-DiR NPs) adhere to MOMC (MOMC/PER-DiR) after intravenous administration. The DiR fluorescence accumulated in the lungs of the MOMC/PER-DiR group, which indicated that compared with MOMC-loading DiR (MOMC-DiR) and free-DiR, MOMC/PER-DiR had a superior ability to target IPF lungs (Fig. 3B). Then, we quantitatively analyzed the drug distribution in the tissues of each organ. The DiR fluorescence intensity in the lungs was 3.5- and 0.5-fold greater than that in the liver in the MOMC/PER-DiR and MOMC-DiR groups, respectively. In addition, there was little accumulation of free DiR in the lungs than that in the liver (Fig. 3C). MOMC loading of DiR could improve DiR accumulation in IPF lungs due to the homing ability of the MOMC.However, the accumulation of MOMC-DiR was weaker than that of MOMC/PER-DiR. This may be because the free dye carried by the MOMC was limited compared with that carried by the PER NPs, which suggested that MOMC/PER could solve the limitation of conventional drug loading of cells. In addition, we further evaluated the homing capacity of MOMC/PER, the responsive release ability of MOMC/PER mediated by MMP-2, the released PER NPs with exposed c(RGDfc), and the retargeting to injured AEC II by immunofluorescence staining. The DiI was chose to a tracer agent, labeling the PER-DiI NPs with red fluorescence, and then, the PER-DiI NPs adhered to MOMC to form MOMC/PER-DiI. Nanog and SPC, which represented MOMC and injured AEC II, respectively, were labeled in green fluorescence. Then, the MOMC/PER-DiI was administered to IPF mice by intravenous injection. As shown in Fig. 3D, the PER-DiI NPs labeled in red overlapped with the MOMC marked in green, generating a merged yellow signal, which revealed that the MOMC/PER-DiI notably accumulated in the lungs of IPF mice in stage 1 (homing to lungs). In stage 2 (releasing PER-DiI NPs), the PER-DiI NPs labeled in red were separated from the MOMC labeled in green, indicating that the PER-DiI NPs were released from the MOMC membrane surface and exposed c(RGDfc) at fibrotic foci as a result of the overexpression of MMP-2 in the IPF microenvironment. Then, the PER-DiI NPs labeled in red overlapped with injured AEC IISPC+ labeled in green, implying that the PER-DiI NPs retargeted to injured AEC II through the interaction between the exposed c(RGDfc) ligand and the v6 receptor on the surface of the injured AEC II in stage 3 (retarget injured AEC II) (Fig. 3D). Collectively, these results showed that MOMC/PER-DiI had the native ability to home to damaged lungs and then were activated by programmed procedures, confirming that MOMC could function as a vehicle to deliver PER NPs to injured lungs.

(A) Schematic of the targeting performance of MOMC/PER in the blood circulation to IPF lungs. (B) In vivo fluorescence images of IPF mice intravenous injection with MOMC-DiR, MOMC/PER-DiR, and DiR (n = 3). (C) Quantification of the in vivo retention profile (n = 3). (D) The different stages of MOMC/PER-DiI. (E) The whole lungs were imaged and investigated after 28 days. Lung morphologies (i) [Photo credit (i): Xin Chang, China Pharmaceutical University], H&E staining (ii), and Masson staining (iii). The morphologies of mitochondria by TEM (iv). The levels of TGF- (F), IL-1 (G), and IL-4 (H) by ELISA assay (n = 5). The levels of lymphocytes (I), white blood cells (J), and neutrophils (K) in whole blood (n = 5). The levels of GSH (L) and SOD (M), respectively (n = 5). (N) The expression of SPC. (O) Survival rate curves (n = 10). Statistical significance was calculated via one-way ANOVA.

To confirm the curative effect of MOMC/PER, we investigated lung morphologies after the administration of MOMC/PER or other treatments. As showed in Fig. 3E, MOMC/PER could greatly relieve IPF according to hematoxylin and eosin (H&E) and Masson staining. Images of lung morphologies showed obvious normalization after treatment with MOMC or MOMC/PER compared with no treatment (Fig. 3E, i). H&E staining showed that lung tissues in the MOMC/PER group were not destroyed and that the alveolar sizes were same as normal lung tissues (Fig. 3E, ii). In addition, compared with no treatment, MOMC also partly protected the lung architecture; however, there was a gap between the MOMC/PER and normal groups. Similarly, Masson staining also showed that the MOMC/PER group exhibited an excellent reduction in collagen I deposition (Fig. 3E, iii). IPF is also induced by mitochondrial oxidative stress in injured AEC II. Hence, we examined the capability of MOMC/PER to repair injured AEC II by maintaining mitochondrial morphologies (Fig. 3E, iv). The morphologies of mitochondria were close to normal in the MOMC/PER group compared with the MOMC group and BLM group, suggesting that MOMC/PER could repair injured AEC II to maintain normal lungs by improving mitochondrial function. Furthermore, we tested the expression of proinflammatory cytokines [TGF-, interleukin-1 (IL-1), and IL-4], which play major roles in excessive ECM formation during IPF progression. As shown in Fig. 3 (F to H), the expression of TGF- in the MOMC/PER treatment group was nearly threefold lower than that in the BLM group, and the expression of IL-1 and IL-4 also decreased by nearly 0.5- and 1-fold, respectively, in the MOMC/PER group compared with the BLM group, suggesting that MOMC/PER could block IPF progression by inhibiting the secretion of proinflammatory cytokines. In addition, the formulations of MOMC and MOMC/PER showed well biocompatibility in a hemolysis test (fig. S5). In addition, inflammatory cells were quantified in whole blood in these groups after treatment. Compared with the BLM group, the MOMC/PER group showed inhibited inflammatory cell proliferation (Fig. 3, I to K), which indicated that MOMC/PER had the ability to alleviate IPF progression in the inflammatory phase. In addition, the results implied that MOMC had a certain ability to inhibit the proliferation of inflammatory cells. Next, glutathione (GSH) and superoxide dismutase (SOD), which are significant inhibitors of ROS, were used to balance the ROS content of injured AEC II. Compared with no treatment, treatment with MOMC/PER increased the GSH level nearly onefold (Fig. 3L), and MOMC also enhanced the GSH level. Similarly, MOMC/PER increased the SOD level to a certain extent in lung tissues (Fig. 3M). We further explored the repair mechanism for injured AEC II in IPF lungs treated with MOMC or MOMC/PER. The expression of SPC was markedly increased in the MOMC/PER group compared with the BLM group; there was also an augmentation in the expression of SPC in the MOMC group, which showed that MOMC/PER could up-regulate AEC II proliferation or recover injured AEC II to normalize the lungs in IPF and demonstrated that MOMC/PER could promote IPF lungs regeneration (Fig. 3N). The survival time of the MOMC/PER group exceeded 60 days, which was longer than the survival time of the BLM group (Fig. 3O), and the MOMC/PER group did not exhibit any changes in body weight (fig. S6).

To investigate the targeting ability of PER NPs through reprogramming to form MOMC/PER in the blood circulation, we conducted the following experiments. The E5-mediated targeting ability of PER-DiI NPs was first evaluated in IPF mice (Fig. 4A). CLSM showed the adhesion of PER-DiI NPs to the surface of MOMC (Fig. 4B, bottom). The confocal images produced the same result as Fig. 2I. Furthermore, PER-DiI NPs were administered to IPF mice model by intravenous injection. The results demonstrated that the PER-DiI NPs adhered to the surface of MOMC (Fig. 4B, middle). More detailed results revealed that the PER-DiI NPs could bind to the MOMC surface and reprogram the MOMC to form MOMC/PER in the peripheral blood by SEM (Fig. 4B, top). In addition, immunofluorescence staining confirmed that the PER-DiI NPs homed to IPF lungs and accumulated in the injured AEC II area after intravenous injection (Fig. 4C). In addition, Nanog-labeled MOMC (green fluorescence) accumulated in higher numbers in IPF lungs than normal lungs, which was similar to previous results (Fig. 2I). We also investigated the targeting capacity of PER-DiR NPs at different time points by in vivo imaging system following intravenous injection, and PLGA-PEG-DiR (PP-DiR NPs), PLGA-PEG-c(RGDfc)DiR (PR-DiR NPs), and PLGA-PEG-E5-DiR (PE-DiR NPs) were used as controls. The PP-DiR NPs and PR-DiR NPs were mainly found in the liver, while the PER-DiR NPs and PE-DiR NPs mainly accumulated in IPF lungs (Fig. 4D and fig. S7A). The accumulation of the PER-DiR NPs in the lungs peaked at 8 hours, while the lung accumulation of the PE-DiR NPs quickly decreased. The primary reason may be that the PE NPs were delivered to the lungs via MOMC; however, they could not anchor on injured AEC II because they lacked c(RGDfc) and were therefore more rapidly cleared from the circulation than the PER-DiR NPs. Compared with the PE-DiR NPs, the PER-DiR NPs accumulated in IPF lungs for a long time (more than 8 hours), which is important for treating lung disease. A quantitative region of interest (ROI) analysis of PER-DiR NPs accumulation was performed by detecting DiR signal variation in the lungs and other organs (Fig. 4E). Moreover, we evaluated PER-DiI NPs behavior in lung tissues after administration at different times (Fig. 4F). After administration at 0.5 hours, increasing levels of overlapping yellow fluorescence in lung blood vessels were observed for PER-DiI NPs labeled in red and MOMC marked in green, indicating that the PER-DiI NPs arrived at IPF lung tissues through reprogramming to form MOMC/PER-DiI in the blood circulation. Then, the red and green signals were separated at the time point of 2 hours, indicating that the PER-DiI NPs were released from the reprogrammed MOMC/PER-DiI due to the overexpression of MMP-2 in the IPF mice. Furthermore, the released PER-DiI NPs showed a wide distribution in the lung tissues at 4 hours after intravenous injection, which is powerful for treating diseases. These data demonstrated that PER NPs could target IPF lungs by means of attaching to circulating MOMC quickly and could accumulate in lung tissues for a long time to achieve therapeutic efficacy.

(A) Schematic of PER NPs circulation in vivo, reprogramming of MOMC/PER, and recruitment to IPF tissue. (B) The targeting ability of PER-DiI NPs. (C) The accumulation of PER-DiI NPs in normal and IPF lungs. (D) Fluorescence IVIS imaging (n = 3). (E) Ex vivo fluorescence imaging and quantification of major organs (n = 3). (F) The accumulation PER-DiI NPs in the lungs at different times. Lung function indexes of GSH (G), SOD (H), and MDA (I). TGF- (J), IL-1 (K), and IL-4 (L) by ELISA assay (n = 5). (M) Proliferation of fibroblasts. (N) Expression of collagen I. Statistical significance was calculated via one-way ANOVA.

We further investigated the antifibrotic efficacy of PER NPs in vivo. With IPF progression, injured AEC II gradually died out due to oxidative stress, which leads to mouse suffocation. Hence, restoring normal lung function has important significance. Compared with the BLM group, the PER NPs group had the promising abilities to repair injured lungs and keep them normal. GSH and SOD levels in the PER NPs group were obviously improved with 0.3- and 1-fold, respectively, which could relieve the oxidative stress in injured AEC II to some extent. The level of malondialdehyde (MDA), a key indicator of oxidative stress, was reduced 0.3-fold in the PER NPs group compared with the BLM group (Fig. 4, G to I). Compared with controls, treatment with PER NPs reduced the production of the three cytokines (TGF-, IL-1, and IL-4) in the lungs by 1-, 0.85-, and 0.7-fold, respectively, which showed that the PER NPs effectively inhibited the inflammatory response in IPF lungs (Fig. 4, J to L). We also examined the levels of TGF-, IL-1, and IL-4 in the spleen tissues (fig. S7, B to D), which showed consistent results. These results indicated that PER NPs could treat IPF by inhibiting inflammatory responses in IPF lungs. As seen in Fig. 4M, immunofluorescence staining results revealed that the population levels of fibroblasts CD90+ labeled in green remained in a relatively stable range, while the population levels of activated fibroblasts indicated great proliferation in the BLM group, supporting the conclusion that compared with no treatment group, the PER NPs had an efficient ability to reverse IPF by inhibiting the activation of fibroblasts. Figure 4N showed that the expression of collagen I was notably decreased in the PER NPs group, which confirmed that PER NPs could achieve therapeutic effects by inhibiting ECM deposition.

We next investigated the antifibrosis mechanism based on the synergistic effect of TRA and AST. We firstly established the different formulations, including PLGA-PEG-TRA-AST (PPTA), PLGA-PEG-TRA (PPT), and PLGA-PEG-AST (PPA), and the morphologies of PPA, PPT, and PPTA were evaluated by TEM (fig S8). As shown in Fig. 5A, the results of immunofluorescence staining showed that the expression of the vimentin as cytoskeletal protein was increased after treatment with different formulations in human lung epithelial cell carcinoma (A549). In particular, compared with other treatments, the PER NPs significantly increased the expression of vimentin, indicating that PER NPs had the capacity to keep injured AEC II normal. To assess the repair mechanism induced by the drugs combination, the ROS were detected using ROS probe 2,7-dichlorofluorescin diacetate (DCFH-DA) via inverted fluorescence microscopy and flow cytometry. The ROS content was significant decreased in the PPTA group compared with the untreated and single-drug groups (PPT and PPA) (Fig. 5B). Although the PPA group exhibited some changes than PPT, this effect was not as strong as that in PPTA group, because the PPTA groups exhibited synergistic effect that relieved oxidative stress in injured AEC II than other control formulations. Furthermore, the PPTA group also showed a reduced mitochondrial membrane potential in TGF-induced cells (Fig. 5C), which supported the conclusion that the efficacy in PPTA group was the result of repairing mitochondrial function with relief of oxidative stress in the mitochondria. In the microenvironment of IPF lungs, myofibroblast can be derived from injured AEC II undergoing epithelial-mesenchymal transition (EMT), which aggravates the progression of IPF. As observed in a wound healing assay and invasion assay (Fig. 5D), PPTA effectively inhibited the occurrence of EMT. Furthermore, fibronectin is a structural protein in the ECM, which is a crucial indicator of IPF progression. The expression of fibronectin was obviously decreased in PPTA group than PPT and PPA groups, thus inhibiting the differentiation of injured AEC II into myofibroblast (Fig. 5E). Next, we also tested IPF-reversing efficacy by monitoring the recovery of the lung architecture and improvement in lung functions in vivo. As shown in Fig. 5F, collagen I deposition in the PPTA group returned to normal levels, as determined by H&E and Masson staining, demonstrating that the PPTA could recover the architecture of injured lungs compared with no treatment or single-drug groups (PPT and PPA). Similarly, the expressions of -SMA and collagen I were tested by immunohistochemistry (IHC), which also obtained the same results that the synergistic effect of PPTA could effectively inhibit myofibroblast activation and ECM deposition. In addition, the level of hydroxyproline, the main component of the ECM, was also decreased after treatment with PPTA compared with other treatments (Fig. 5G), suggesting that PPTA had the ability to diminish ECM deposition and retard IPF progression. Furthermore, we detected the expression of -SMA to evaluate the myofibroblasts activation by Western blotting. The lungs were collected after treatment with PPTA, PPT, or PPA for 28 days. The results showed that -SMA expression, as the major evaluation index for IPF, was significantly reduced in PPTA group (Fig. 5H). In addition, the results of real-time quantitative polymerase chain reaction (qPCR) showed that the relative mRNA expressions of CTGF (Ctgf) and -SMA (Acta2) significantly decreased in PPTA group, which indicated that the combination of AST and TRA can achieve efficient therapeutic efficacy by inhibiting myofibroblasts overactivation (Fig. 5, I and J). Moreover, MDA expression decreased, and SOD and GSH levels increased after treatment in PPTA group compared with the PPT and PPA groups. Together, these results implied that the combination of AST and TRA could recover IPF lung function through synergistic effect that was not observed with the other treatments (Fig. 5, K to M). The various formulations as mentioned above were safe by intravenous injection through H&E staining (fig. S9).

(A) Expression of the vimentin in vitro. (B) The ROS level in vitro. (C) The changes of mitochondrial membrane potential. (D) Invasion assay. (E) Fibronectin expression. (F) H&E, Masson, and IHC staining. (G) The level of hydroxyproline. (H) The -SMA and -actin by Western blotting. The mRNA expression of Acta2 (I) and Ctgf (J) by qPCR (n = 3). Contents of GSH (K), MDA (L), and SOD (M) (n = 5). Statistical significance was calculated via one-way ANOVA.

To further investigate the antifibrotic efficacy of MOMC/PER and pirfenidone as a conventional therapeutics for IPF, we evaluated the ability of these treatments to repair lung tissue and inhibit collagen I deposition through H&E and Masson staining, respectively, after 28 days of administration. As shown in Fig. 6A, the alveolar structure in the BLM group collapsed, and alveolar wall thickness increased notably, indicating that collagen I was accumulated and that alveolar heterogeneity was aggravated. Similarly, the alveolar morphologies in the pirfenidone group also showed collapse via H&E staining. In contrast, MOMC/PER could obviously repair the collapsed part of the alveolar space, narrow the spaces between the alveoli, and produce a thinner alveolar wall that tended to appear normal by H&E staining, which demonstrated that MOMC/PER had greater reparative effect on alveolar structure than pirfenidone. In addition, MOMC/PER group showed notable decrease compared with the pirfenidone group in inflammatory cell infiltration. The PER NPs were also more competent in restoring alveolar structure than the clinical drug pirfenidone. This effect was observed because the PER NPs could undergo reprogramming to form MOMC/PER in the blood circulation and then reach their destination, which was consistent with the above results. We further confirmed therapeutic efficacy in regard to ECM deposition by Masson staining (Fig. 6B). Compared with that in the pirfenidone group, the ECM accumulation in the lungs, which appeared as blue staining, was notably reduced in the MOMC/PER group. The results for Masson staining showed that MOMC/PER had greater power than pirfenidone to prevent IPF progression by inhibiting ECM deposition. The main reason for the limited therapeutic effect of pirfenidone was its low bioavailability as an oral drug, and onefold treatment target is the second therapeutic limitation of pirfenidone. Overall, the antifibrotic efficacy in the MOMC/PER group was the best efficacy observed through H&E and Masson staining, and PER NPs had better efficacy than pirfenidone or MOMC. In addition, the fibrosis score of different formulations showed the same trend in fig. S10. These data demonstrated that MOMC/PER showed a preferable combination efficacy over the U.S. Food and Drug Administration (FDA)approved therapeutic pirfenidone or using MOMC or PER NPs alone.

(A) H&E staining. (B) Masson staining. The levels of TGF- in the lungs (C) and spleen tissues (D). N.S., not significant. The levels of IL-1 in the lungs (E) and spleen tissues (F). BUN (G), ALT (H), and aspartate aminotransferase (I) in serum (n = 4). Statistical significance was calculated via one-way ANOVA.

Then, we further evaluated the antifibrotic effect in various groups by examining biochemical indexes of IPF. We first examined the expressions of TGF- and IL-1 in the lungs and spleen tissues, respectively. The results in the lungs showed that TGF- expression was reduced onefold in the MOMC/PER group (P = 0.015) compared with the BLM group and became close to normal (Fig. 6, C and D). However, there was no significant difference between the pirfenidone group and the BLM group, and the level of TGF- in the PER NPs group was lower than that in the pirfenidone group. In addition, the TGF- level in the spleen tissues was significantly decreased in the MOMC/PER group (P = 0.002) compared with the BLM group. However, the level of TGF- in the pirfenidone group was similar to that in the BLM group. The main reason is that pirfenidone is used to treat IPF by inhibiting the accumulation of collagen I, but it has no therapeutic effect on the simultaneous inflammatory response or cytokine expression. The results demonstrated that MOMC/PER had the best antifibrotic efficacy, which was superior to the efficacy achieved by pirfenidone and was mediated by inhibiting the expression of cytokines in the lungs. Furthermore, we detected the expression of IL-1 in the lungs and spleen tissues to investigate the antifibrotic effects of different formulations (Fig. 6, E and F). The trends in IL-1 expression were similar to TGF-; all the treatments could reduce the expression of IL-1, and MOMC/PER showed the best therapeutic efficacy (P = 0.001) in all the treatments. In addition, the IL-1 level in the pirfenidone group was maximal, indicating that compared with the other treatment groups, including the MOMC/PER and PER NPs alone groups, the pirfenidone group showed minimal anti-inflammatory effects. These results indicated that MOMC/PER could achieve a greater treatment effect on IPF than pirfenidone by inhibiting the expression of cytokines in the inflammatory phase; the efficacy of PER NPs was second only to MOMC/PER, and pirfenidone and MOMC were weaker than the PER NPs.

To assess the safety of the treatments in vivo, we then evaluated biological indexes for each formulation after treatment. The levels of blood urea nitrogen (BUN), alanine transaminase (ALT), and aspartate aminotransferase were detected to evaluate the function of the kidneys, liver, and heart, respectively (Fig. 6, G to I). The levels of BUN, ALT, and aspartate aminotransferase were not significantly different between the pirfenidone and other groups (the MOMC/PER, PER NPs, and MOMC groups). As an oral drug approved by the FDA for the treatment of IPF, pirfenidone is highly recognized for its safety in application. Similarly, our different formulations obtained results of safety equivalent to pirfenidone for a certain period of time, indicating that MOMC/PER, MOMC, and PER NPs could also be safely administered by intravenous injection and could be used clinically.

IPF is characterized by injured AEC II and activated myofibroblast, resulting in ECM deposition. To date, the FDA has approved only two drugs (pirfenidone and nintedanib) for IPF treatment. Unfortunately, curing end-stage IPF is inefficient due to the narrow therapeutic spectrums and insufficient accumulation of these drugs in the lungs (3). As a result, traditional therapies have done little to reverse IPF (4). To address this problem, we developed programmed therapeutics MOMC/PER to reverse IPF by efficient lung delivery, programmed modules, and double synergetic strategies.

Two synergetic strategies including drug/drug and cell/drug involved in reversing IPF were shown for the MOMC/PER here. First, the drug/drug as weeding and uprooting strategy could repair injured AEC II and inhibit myofibroblast activation, achieving first synergetic antifibrosis effect. In particular, one drug (AST) acted as the uprooting part of the treatment strategy, repairing injured AEC II by neutralizing oxidative stress. The other drug (TRA) acted as the weeding portion of the strategy, inhibiting the differentiation of fibroblasts into myofibroblast by suppressing CTGF production. Second, some studies have demonstrated that MOMC is multipotent cell that can be specifically recruited to injured lung tissues through interactions between chemokine receptors and chemotactic factors (3638) and contribute to lung tissue normalization and regeneration (39). In addition, MOMC also plays a vital role in regulating the population of immune cells during the inflammatory phase of disease progression (40). Similarly, our results also showed that MOMC could inhibit the proliferation of inflammatory cells, such as lymphocytes and white blood cells. This is another synergetic effect called cell/drug. PER NPs also exhibited greater antifibrotic effects than pirfenidone due to their efficient lung-targeting ability and combination of AST and TRA, as these PER NPs could target MOMC in the circulation, accumulate in the lungs effectively, and then reverse IPF collaboratively. The limited treatment efficacy of pirfenidone is mainly due to its low bioavailability, narrow therapeutic spectrum, and functions by inhibiting myofibroblast activation only.

In addition, MOMC/PER is strategically distinct from nanodelivery carrier (41) and drug-loaded cells carrier (14). The traditional nanodelivery system for IPF always presents dissatisfactory accumulation and unexpected drug release at the lesion site. Even these defects can be avoided for IPF therapy, the therapeutic efficacy is also limited to onefold treating target, and these shortcomings make IPF hard to reverse. In addition to nanodelivery systems, cell-mediated drug delivery has also received more attention in disease treatment. The classic cell-based delivery strategy for treating disease is reliant on drugs being loaded into cells by endocytosis (15). However, cells are difficult to load with large quantities of drugs, and chemotherapeutics may be highly toxic to cells undergoing loading. Hopefully, the PER NPs adhere to the MOMC surface in our study could surmount this challenge in conventional drug loading of cells. PER NPs were firstly attached to MOMC surface and then precisely delivered to the lungs via the homing ability of MOMC and activated for IPF reversion. However, there is still an unresolved point in our research, which is that the treatment mechanism of MOMC remains unclear. Our results indicated that MOMC might up-regulate AEC II proliferation or recover injured AEC II to normalize the lungs. The mechanism of MOMC differentiation for IPF treatment requires further exploration. In addition, some studies have indicated that MOMC could partly treat early IPF through regulating the immune response by inhibiting the proliferation of immune cells in vivo (42). It is unclear whether MOMC is effective for IPF therapy during different periods.

Compared with conventional antifibrotic strategies, our previous unknown programmed therapeutics MOMC/PER has showed accurate lung targeting and excellent therapeutic effects. The excellent antifibrotic efficacy of the MOMC/PER was achieved through the following features. (i) MOMC has the ability to backpack PER NPs, constructing programmed therapeutics MOMC/PER. (ii) MOMC/PER can precisely accumulate in IPF lung tissues due to the homing ability of the MOMC. (iii) PER NPs are sensitively released from MOMC/PER due to the overexpression of MMP-2 in the IPF microenvironment. (iv) Released PER NPs are able to retarget injured AEC II through c(RGDfc). (v) PER NPs can reduce the secretion of TGF- by occupying TGF-latent sites. (vi) Two drugs loaded into PER NPs are the key factors in achieving IPF reversion of drug/drug as weeding and uprooting. In addition, MOMC also participates in AEC II regeneration using cell/drug strategy. Specifically, MOMC-mediated delivery therapeutics is convenient, and the materials used in our PER NPs have all been approved by the FDA, which indicates certain advantages for further clinical development. Overall, we have proposed an innovative concept to cure IPF through using native cells as a delivery carrier and a dual-drug combination as therapeutic agents, and this strategy is likely to be applicable to other major diseases.

MMP-2, -SMA rabbit anti-mouse antibody, and DCFH-DA were purchased from Sigma-Aldrich (St. Louis, USA). SPC rabbit anti-mouse antibody was purchased from Millipore (St. Louis, USA). Lymphocyte isolation kit was purchased from Solarbio Science & Technology Co. Ltd. (Beijing, China). PLGA-PEG-Mal and PLGA-PEG-c(RGDfc) were purchased from Jinan Daigan Biomaterial Co. Ltd. (Jinan, China). RPMI 1640, fetal bovine serum (FBS), and bicinchoninic acid (BCA) protein assay kit were purchased from Jiangsu KeyGEN BioTECH Co. Ltd. (Nanjing, China). The peptide E5 (CGPLGIAGQCGGRSFFLLRRIQGCRFRNTVDD) was synthesized by Top Peptide Biotechnology Co. Ltd. (Shanghai, China). AST was purchased from Yuanye Bio-Technology Co. Ltd. (Shanghai, China). TRA was purchased from J&K Scientific Co. Ltd. (Beijing, China). DiI, 3,3-dioctadecyloxacarbocyanine perchlorate (DiO), and DiR were purchased from Fanbo Biochemicals Co. Ltd. (Beijing, China). DAPI (4,6-diamino-2-phenylindole) and mitochondrial membrane potential kit of JC-1 were purchased from Beyotime Biotechnology Co. Ltd. (Shanghai, China). CXC chemokine ligand 12 (CXCL 12) and CC chemokine ligand 19 (CCL 19) were purchased from Zoonbio Biotechnology Co. Ltd. (Beijing, China). BLM was purchased from Zhejiang Huahai Pharmaceutical Co. Ltd. (Linhai, China). TGF- was purchased from Multi Sciences Biotech Co. Ltd. (Hangzhou, China). C6 was purchased from Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan). LysoTracker Red DND-99 kit was purchased from Thermo Fisher Scientific (Waltham, USA). Nanog, TGF-/Smad and collagen I rabbit anti-mouse antibodies, and H&E and Masson staining kits were purchased from Servicebio Co. Ltd. (Nanjing, China). GSH, MDA, SOD, and hydroxyproline were purchased from Jiancheng Biotech Co. Ltd. (Nanjing, China). IL-1 and IL-4 detection kits were purchased from eBioscience (Waltham, USA). TGF- detection kit was purchased from BioLegend (CA, USA). Polyvinylidene difluoride (PVDF) was purchased from PALL (NY, USA). Electrogenerated chemiluminescence (ECL) was purchased from Tanon Science & Technology Co. Ltd. (Shanghai, China).

MOMC was isolated from peripheral blood by a mouse lymphocyte isolation kit. To obtain the MOMC, peripheral blood was collected from the C57BL/6J mice of IPF with EDTA and diluted three times with phosphate-buffered saline (PBS). The isolated protocol of MOMC was as follows: The mouse Percoll was added into diluted peripheral blood solution, and MOMC was isolated from peripheral blood by centrifugation at 600 rpm for 30 min. The solution was divided into upper, middle, and lower layers, and MOMC existed in the middle layer. Then, the MOMC was taken into centrifuge tube of 15 ml, and cell washing buffer of 5 ml was added into the tube. The cell suspension was then centrifuged for another 30 min, and it needed to be repeated three times to obtain MOMC. Last, the cell deposits were resuspended, and the suspension was put into the culture dish in RPMI 1640 medium with 10% FBS at 37C and 5% CO2 (22, 23). The adherence time for dishes of MOMC was about 2 weeks. The MOMC was a kind of adherent cells, and the morphologies of cells were fusiform. After the cells adhere to the dish, the medium was changed once every 3 days.

PER NPs were prepared by antisolvent precipitation method. Peptide E5 modification was prepared as follows. Specifically, the PLGA-PEG-Mal and PLGA-PEG-c(RGDfc) (mass ratio, 10:1) and dual drugs of AST and/or TRA were dissolved in dimethyl sulfoxide (50 mg/ml, 2 ml), added dropwise into deionized water with 100 ml, and then stirred with 300 rpm for 2 hours. Next, the prepared nanoparticle solution (NPs) was centrifuged at 2800 rpm for 15 min to discard the large particles and free drug. The NPs were then condensed to concentration of 2 ml by ultrafiltration device for further use. Last, the peptide E5 was added into the solution of NPs to form PER NPs. The preparation process of other groups including PLGA-PEG-c(RGDfc) (PR NPs), the preparation of PLGA-PEG-E5 (PE NPs), and the preparation of PLGA-PEG (PP NPs) were similar to PER NPs.

The preparation of MOMC/PER was carried out by incubating MOMC with PER NPs. Briefly, the MOMC (2 105 cells/ml) was cultured in a petri dishes with a diameter of 100 mm. After incubated with the FBS-free media for 1 hour, PER NPs at a TRA concentration of 40 g/ml were added into MOMC medium and incubated for 2 hours at 37C and 5% CO2. At the same time, the CXCR4 receptor and ligand peptide E5 would undergo bioconjugate reaction.

The hydrodynamic diameters and potentials for PER NPs suspended in 1 PBS were measured by Brookhaven Instruments (NY, USA). The morphologies of PER NPs were characterized by TEM (Hitachi TEM system, Japan).

For MOMC/PER-DiI, characterization of adhesion between MOMC and PER-DiI NPs was imaged by CLSM (Carl Zeiss 700, Germany). Specifically, MOMC was cultured in 35-mm culture dishes and incubated with PER-DiI NPs for 2 hours, and the preparation of PER-DiI NPs was the same as mentioned above. The nucleus of MOMC was labeled with DAPI and MOMC membrane was labeled by DiO. For SEM characterization, MOMC/PER-DiI was coated with gold/palladium and examined by Hitachi-SU8020 (Japan).

The sensitive release properties of PER NPs from MOMC/PER in vitro was evaluated under the microenvironment of MMP-2 in vitro. MMP-2 enzyme was applied to release PER NPs by degrading the linker of GPLGIAGQ between PER NPs and MOMC. The characterization of released activity was investigated in vitro. First, MMP-2 (2 g/ml, 1 ml) was added into MOMC/PER-DiI medium for 30 min. Then, MOMC and released PER-DiI NPs were detected by flow cytometry (BD FACSCalibur, USA) or fixed by 4% paraformaldehyde (w/v) for CLSM and 2.5% glutaral for TEM. Besides, the nucleus of MOMC was labeled with DAPI for 30 min at 37C, and the MOMC membrane was labeled with DiO for 15 min at 37C. MMP-2 enzyme was dissolved in deionized water and free RPMI 1640 (volume ratio, 1:20) at a concentration of 2 g/ml. After that, the released PER NPs and MOMC in solution were prepared for the other testing assay and image.

The loading ability of MOMC was detected by the concentration of TRA and AST. First, MOMC was cultured in dishes with 100 mm. After MOMC adhered on the dishes, PER NPs were added into the culture dishes and incubated with MOMC for 2 hours in RPMI 1640 with free FBS, and then, MOMC/PER was washed thrice with PBS and digested with 0.25% trypsin-EDTA solution. Next, cells were harvested by centrifugation at 1000 rpm for 3 min and counted with a hemocytometer. Last, the cells were resuspended with PBS of 1 ml, and the absorbance was determined at 326 nm for TRA and 491 nm for AST by Multiskan GO (Thermo Fisher Scientific, USA).

The migration capacity of MOMC/PER was investigated by a Transwell device. First, the MOMC were cultured into the upper chambers with pore sizes of 8.0 m with 2 104 in 400 l of RPMI 1640 with FBS-free media for 24 hours. The cells were divided into three groups, including CXCL 12 () or CCL 19 () of MOMC, CXCL 12 (+) or CCL 19 (+) of MOMC, and CXCL 12 (+) or CCL 19 (+) of MOMC/PER. RPMI 1640 with 10% FBS and CXCL 12 (10 g/ml) or CCL 19 of 600 l was added to the lower chamber into the 24-well plates. Then, the MOMC in MOMC/PER group was added PER NPs at concentration of 40 g/ml and incubated for another 24 hours. At last, the Transwell chambers were stained with crystal violet and were dissolved with 33% acetic acid, and the absorbance of solution was tested at 570 nm.

C57BL/6J male mice were obtained from East China Normal University Laboratory Animal Technology Co. Ltd. (Shanghai, China) and housed with a 12-hour light/12-hour dark cycle at 25C. All the animal protocols and procedures were performed under the guidelines for human and responsible use of animals in research approved by the regional ethics committee of China Pharmaceutical University. After acclimatization for 7 days, mice were subjected to IPF model experiments. IPF mice models were established by inhalation of BLM through endotracheal intubation (2 U/kg, 40 l; Braintree Scientific, USA). Next, the mice were randomly assigned to the treatment. For the cell culture, A549 and MOMC were cultured in RPMI 1640 media containing 10% FBS and 1% penicillin and streptomycin at 37C and 5% CO2.

A549 cells were cultured in six-well plates at 37C and 5% CO2 for 24 hours and then incubated for another 24 hours with pure PPA, PPT, or PPTA at TRA concentration of 20 nM. The expression of vimentin and fibronectin was investigated by immunofluorescence staining.

The invasion ability of TGF-induced A549 cells was evaluated by wound healing tests after treated with various formulations. First, A549 cells were seeded in six-well plates at 15 104 cells per dish and incubated for 24 hours. Next, a 10-l pipette tip was used to scratch wells in the middle of the dishes, and then, A549 cells were washed three times with PBS to remove suspended cells. The cells from each group were imaged by inverted fluorescence microscope (Nikon, Japan) to observe the extents of wound healing after treated with PPT, PPA, PPTA, and PER NPs at 24 hours.

The migration capability of TGF-induced A549 cells was investigated by a Transwell device. The A549 cells of 5 104 in 400 l of RPMI 1640 with FBS-free media were added to the upper chambers with pore sizes of 8.0 m for 24 hours, and RPMI 1640 with 10% FBS media of 600 l was added to the lower chamber into the 24-well plates. Then, the cells were incubated with PPT, PPA, PPTA, and PER NPs (20 nM of TRA concentration) for another 24 hours. After incubation, the Transwell chambers were stained with crystal violet and were dissolved with 33% acetic acid, and the absorbance of solution was tested at 570 nm.

A549 cells were seeded on six-well plates (15 104 per well) and incubated overnight. First, the cells were activated by TGF-, and then, various treatment groups were incubated with A549 cells for 24 hours (TGF-, normal, PPT, PPA, PPTA, and PER NPs). Next, the ROS probe DCFH-DA (5 M) was added into the dishes and incubated with cells in RPMI 1640 media with free FBS at 37C under 5% CO2 in the dark for 15 min. Last, cells were digested, and the content of ROS was analyzed by flow cytometry (BD Accuri C6, USA) and imaged by inverted fluorescence microscope, respectively.

A549 cells were cultured on six-well plates (15 104 per well) overnight at 37C and 5% CO2. After treated with different formulations for 24 hours, 500 l of mitochondrial membrane potential reagent of JC-1 (1) solution was added into the dishes for 20 min. Then, the cells were stained by 4% paraformaldehyde (w/v) and imaged by inverted fluorescence microscope.

First, A549 cells were seeded on a 35-mm sterile glass bottom culture dishes (2 105 cells) and cultured overnight in RPMI 1640 with 10% FBS. The preparation of PR-C6 and PP-C6 was same as mentioned above for PER NPs. The PR-C6, PP-C6, and C6 were then incubated with A549 cells for 4 hours at 5 g/ml. Next, the cells were washed three times by PBS, and the nucleus was stained with DAPI. Images and data were acquired with CLSM and flow cytometry (BD Accuri C6, USA).

The A549 cells of 5 104 were cultured in a 35-mm sterile glass bottom culture dishes. After the cells were cultured overnight in RPMI 1640 with 10% FBS for 24 hours, PR-C6 and PP-C6 were incubated with A549 cells for 1 and 4 hours at 5 g/ml. Then, the cell dishes were washed three times by PBS and fix by 4% paraformaldehyde (w/v). Lysosome was stained with LysoTracker Red DND-99 kit (100 nM) for 15 min, and the dishes were washed three times with PBS. Then, the cell nucleus was stained by DAPI like above method. Images were acquired by CLSM.

MOMC/PER-DiR was prepared by the method as mentioned above for PER NPs. The homing ability of MOMC was detected in vivo by IVIS imaging system (Kodak, USA). C57BL/6J mice were induced IPF by inhalation of BLM. Then, the IPF mice were injected with MOMC/PER-DiR, MOMC-DiR, and free DiR by intravenous injection and tested by IVIS living system at different point times. Then, the mice are sacrificed, and the lungs and other organs were harvested for ex vivo imaging after 8 hours of intravenous injection. ROI was circled around the lungs and the other organs (liver, heart, spleen, and kidneys). The fluorescence intensity of the DiR was determined by living image software.

The PER NPs can be released from MOMC/PER-DiI by MMP-2, owing to responsive blocking the linker between PER NPs and MOMC. We have investigated the homing capability, responsive release, and retarget ability. We first administrated MOMC/PER-DiI by intravenous injection, and the preparation of MOMC/PER-DiI was applied by the methods as mentioned above. The mice were sacrificed, and lung tissues were harvested at different point time of 30 min, 1 hour, and 2 hours in a dark place. First, the lung tissues were fixed with 4% paraformaldehyde (w/v), and then, the tissues were embedded and dewaxed before slicing. Next, the lung slices were labeled by Nanog and SPC at 4C for 1 hour and washed with 0.2% Triton X-100 for three times. Then, they were incubated with relevant secondary antibodies for 2 hours. Thereafter, the slices were stained with DAPI and viewed under fluorescence microscope.

The IPF mice were injected with PER-DiR NPs, PE-DiR NPs, PR-DiR NPs, and PP-DiR NPs by intravenous injection at different point times for 1, 4, 8, 12, and 24 hours and tested by IVIS imaging system. The preparation of PER-DiR NPs, PE-DiR NPs, PR-DiR NPs, and PP-DiR NPs were the same method as mentioned above, and then, the mice were sacrificed, and the lungs and other organs were harvested to detect ex vivo imaging after intravenous injection at 24 hours. ROI was tested on the lungs and the other organs (liver, heart spleen, and kidneys). The fluorescence intensity of the DiR was determined by living image software.

The antifibrotic efficacy in MOMC/PER, MOMC, PER NPs, and pirfenidone for IPF treatment in vivo was evaluated on IPF male mice (C57BL/6J, age of 6 to 8 weeks). The preparations of MOMC/PER and PER NPs were the same method as mentioned above. Pirfenidone was administered by gastrointestinal because it is an oral medication, and other formulations were administered via intravenous injection.

The contents of MDA, SOD, and GSH were detected after treated with various treatments in lung tissues. The protocols are as follows: Solution 1 of 1.5 ml was added into the lung tissues solution (0.5 ml) and mixed thoroughly. Then, the samples were centrifuged for 10 min at 3500 to 4000 rpm. The sample supernatant was added into 3,3,5,5-tetramethyl benaidine (TMB substrate), and the absorbance of GSH was detected after 5 to 10 min at 420 nm. The contents of SOD and MDA were tested at 550 and 532 nm, respectively.

The lungs and spleen tissues were collected and diluted in precooled solution. The IL-1, TGF-, and IL-4 in lung tissues were assayed using enzyme-linked immunosorbent assay (ELISA) method as instructed by the manufacturer. First, the wells were washed three times with a washing buffer for 3 min each time. Next, the blocking solution of 200 l was added into each well and incubated for 1 to 2 hours at 37C. Then, the sealing film was removed carefully and putted it into the washing machine and washed three to five times. Furthermore, the sample of 100 l was added and should be tested diluted appropriately to the above coated reaction wells, and the diluted biotinylated antibody working solution was added with 100 l into each well. Then, the samples were sealed with a sealing membrane and incubated at 37C for 1 hour. The following step was that 100 l of diluted enzyme conjugate working solution was added into each well. Next, TMB substrate solution with 100 l was added into each well and should be avoided reaction with light for 10 to 30 min at 37C until a notable color gradient appears in the diluted standard well. Within 10 min, the absorbance of each well was measured on a microplate reader at 450 nm with zero adjustment of the blank control well.

The content of hydroxyproline was detected by a hydroxyproline detection kit. The lung tissues of 30 to 100 mg were mixed with hydrolysate to 1 ml and hydrolyzed in boiling water for 20 min. The sample solution was pH 6.0 to 6.8. Then, the serum hydrolysate was added activated carbon and mixed at 60C for 15 min. After cooling, the serum samples were centrifuged at 3000 rpm for 20 min, and the supernatant was detected by microplate reader at 550 nm.

The whole blood from mice was collected by anticoagulant tube with EDTA, and white blood cell counts (including lymphocytes and monocytes) were assayed using a standard blood analyzer (Mindray, China).

After treatment with different formulation, the lungs, heart, liver, spleen, and kidneys were harvested, and lungs were investigated by H&E, Masson, and IHC staining. Other organs were detected by H&E staining to evaluate the application security. First, the lung tissues were fixed with 4% paraformaldehyde (w/v) for more than 48 hours and embedded in paraffin. Then, the lung tissues were cut into 4-m sections for H&E staining and Masson staining. The levels of collagen I and -SMA were evaluated by IHC, and protocols are as follows: The slices were incubated with primary antibodies (collagen I and -SMA or fluorescently labeled CD90 and collagen I, Servicebio, China) and then incubated with corresponding secondary antibodies to detect the expression of relative proteins.

The qPCR analysis was evaluated RNA expression of Ctgf and Acta2. qPCR was conducted in ABI StepOnePlus (Thermo Fisher Scientific, USA). Lung tissues (100 mg) were homogenized to extract the total RNA according to the protocols. Complementary DNA (2 g) was prepared using the Reverse Transcription System, and then, the expression of related genes was determined using q-PCR. The primers used are Acta2 (NM_007392.3), Ctgf (43), and Gapdh (NM_008084.2).

The harvested lungs were homogenized in PBS buffer and then centrifuged at 3000 rpm for 30 min. The supernatant of total protein was taken for further experiments. Total protein concentration in the solution was determined with a BCA protein assay kit. After detecting in SDS-PAGE with protein samples in different treatment groups, the bands were transferred onto PVDF membrane. Next, the PVDF membranes were blocked with 5% milk at room temperature for 2 hours and incubated with primary antibodies (-SMA, TGF-/Smad, and -actin rabbit anti-mouse antibodies) at 4C overnight and then incubated with corresponding secondary antibodies for 2 hours at room temperature. Last, the bands were detected using ECL (Tanon, China) Western blotting substrate (Thermo Fisher Scientific, USA). The -actin was used as an endogenous control.

The mice were sacrificed, and the serum samples were detected in different treatment groups. The contents of ALT, aspartate aminotransferase, and BUN in the serum were determined using the relevant assay kits (Servicebio, China).

Statistical analyses were performed using GraphPad Prism software (GraphPad Software, USA). All error bars were means SEM; differences detection index between the treated groups and control groups were determined via one-way analysis of variance (ANOVA). P < 0.05 was considered significantly different.

Acknowledgments: We thank the Cellular and Molecular Biology Center of China Pharmaceutical University for assistance with confocal microscopy work. Funding: This work was supported by the National Key R&D Program of China (2017YFA0205400). We thank the National Natural Science Foundation of China (NSFC; grant nos. 81773667, 81573369, and 81430082) and NSFC Projects of International Cooperation and Exchanges (81811540416). This work was also supported by the Fundamental Research Funds for the Central Universities (2632018PT01 and 2632018ZD12), the 111 Project from the Ministry of Education of China and the State Administration of Foreign Experts Affairs of China (B16046), the Double First-Class Project (CPU2018GY06), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions. Author contributions: H.-L.J., H.-P.H., X.C., and L.X. conceived the project, designed all the experiments, analyzed the data, and wrote the manuscript. X.C. and L.X. conducted the experiments. X.C. and Y.W. analyzed the data. X.C., Y.W., C.-X.Y., Y.-J.H., T.-J.Z., X.-D.G., and L.L. wrote the manuscript. All authors edited the manuscript. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.

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Monocyte-derived multipotent cell delivered programmed therapeutics to reverse idiopathic pulmonary fibrosis - Science Advances

Recommendation and review posted by Bethany Smith

Victoria cave researcher, Genevieve von Petzinger,is suddenly ina pretty good positiontofind what she calls the Holy Grail of her field.

The University of Victoriapaleoanthropologist, who specializes in European cave art, has been awarded a National Geographic grant to test genetic material found in cave wall paint in Spain to try tofind out who forgot to sign their work at least 40,000 years ago.

A DNA test, which would reveal genetic mutations due to evolution, could help pinpoint the time perioda painting was made and may helpdetermine if the art was actuallythe handiwork of humans or Neanderthals who lived about130,000 to 40,000 years ago.

"'It would just be so fascinating to see the identity," said PetzingerTuesday on On The Island."The million dollar question is, did Neanderthals paint?"

And there is already someindication, according to von Petzinger, thatthis extinct species was, in fact, artistic.

Von Petzingersaid, a few years ago, some of her colleagues tested samples of minerals they foundcovering cave drawings and determined the minerals to be 65,000 years old, which von Petzinger said indicated the art underneath was older and, therefore, drawn byNeanderthals.

But, she said, this dating methodwas hotly debated by others in the field.

"It was quite the big drama going back and forth," said von Petzinger, who then thought maybe genetic testing could bea way to get a definitive answer.

Genetictesting can even pinpoint the artist's gender and, saidvon Petzinger, possibly lead to findinga living descendent.

"This would be a world first," she said, her voice brimming with excitement.

Von Petzingersaid she is incredibly grateful to colleagues, the Spanish government and National Geographic for being willing to believe in her "crazy idea."

But it's not without precedent.

She said other researchers havehad success testing genetic material found in dirt on the floor of caves in Croatia and she is "cautiously optimistic"she will have similar success with paint substances.

There is a bit of a snag though.

The money is in place, and all the players are on board, but it could be awhile before von Petzinger can board a plane.

She said her tentative plan is to be back in the caves, where she previously spentasignificant amount of time doing field work, by summer 2021.

The COVID-19 pandemic has thrown a wrench into field work for many academics, but von Petzinger said she is staying connected with colleagues online andit isa great time for scientists to slog through statistical analysis work they might as well get done while they can't get out.

And it could be because of her line of work, but von Petzingeris cautiously optimistic about the current state of the world aswell.

"This pandemic is certainly a pretty scary thing, but our ancestors have survived some pretty scary things themselves," she said. "Our species is very resilient."

To hear the complete interview withGenevieve von Petzingeron On The Island tap here.

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Did Neanderthals draw? This B.C. researcher is going to test DNA in old cave art to find out - CBC.ca

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Ancestry and 23andMe offer direct-to-consumer DNA tests.(Photo: Reviewed.com/Jackson Ruckar)

It's a question that has vexed researchers from the beginning of the coronavirus outbreak: Why do some people get severely ill and die from COVID-19, while others have mild symptoms or none at all?

Now, scientists at two direct-to-consumer genealogy DNA companies hope to use the genomesthey've collected from millions of people over the years to see if they can find a genetic explanation toanswer that question.

Both 23andMe and Ancestry have launched COVID-19 studies, asking U.S. adult customers who've already submitted DNA samples to answer online questions about how the virus affected or didn't affect them.

"From the early days ... I think it was clear to all of us that some people were getting very, very sick when they were affected with coronavirus, and some people had hardly any symptoms at all," said Dr. Catherine Ball,chief scientific officer at Utah-based Ancestry."It turns out that there are plenty of people who have no symptoms. The spectrum of human response to the same pathogen is unusual.

"And even with a bunch of comorbidities and other problems, it's still remarkably divergent in different people, even if they have the same age and have the same overall health. And soto geneticists, that looks like there's a genetic factor in whether people become infected in the first placeor have serious or mild symptoms."

With 16 million people who've already spit in vials and sent them to Ancestryfor genetic testing to find blood relatives who might be closely or distantly related or learn how much of their DNA suggests their relatives came from Africa or Asia or were Native American or European, Ball said the companyknew it had a potentially useful data pool to tap for COVID-19 research.

"We clearly want to take the opportunity to unleash that power to be able to see if there are genetic signals, and be able to help researchers and people making drugs and therapeutics and vaccines dosmarter work faster," she said.

Dr. Catherine Ball, chief scientific officer for Utah-based Ancestry.(Photo: Ancestry)

Of those 16 million DNA customers through Ancestry, so far about 500,000people have already taken an online survey to participate in the company'scoronavirus research.

At 23andMe, principal scientist Adam Auton said the California-based company's COVID-19 genome-wide association study launched in April.

About 10 million of itsgenotype customers are eligible for the study, he said.Of them, about 80%have consented to participate in research, and600,000 customers have opted into the COVID-19 study.

"It is a really quite tremendous response to the study and I think shows that people really do want to try and contribute to help understand and fight this disease," said Auton.

Both Ancestry and 23andMeacknowledge that the bigger the sample size, the better their research will be.

"Never ask a scientisthow much data she needsbecause she always needs more," Ball said. "We're really hoping to get a minimum of a million respondentsbecause we need to have a decent number of people who have tested positive to give us a statistical signal."

So far, about9,000 people in 23andMe'sCOVID-19 study reported that theytested positive for coronavirus.

"That's a pretty substantial number," Auton said. "However, it's the nature of genetic studies that we really need very large numbers of people to be able to draw connections between the genetic information and people's health information."

A worker at 23andMe performs DNA testing on samples provided by customers.(Photo: 23andMe)

Since the pandemic began,about 1.6million people in the United States, a country of 330 million, have tested positive for COVID-19. As the virus continues to spread,and more people get coronavirus diagnoses, the companies suspect that the number of people who will go on to enroll in their studies also is likely to rise.

"We understand this is an evolving situation," Ball said. "And while we can't shelter in place forever, at some point, as we're opening up our cities and states, more people will start contracting the virus."

Anyone who may have already filled out anonline COVID-19 survey on Ancestry.com or 23andMe.com, saying they had not yet had the virus, can go back and revise their answers later to reflect that they've contracted it.

To expand its research of people who've had COVID-19 even more, Auton said 23andMeis now offering tomail a free DNA test kit to any U.S adult who was hospitalized with COVID-19, but has not yet submitted a DNA sample to the company.

"We are essentially asking if people have been hospitalized with COVID-19, and they have recovered, if they would like to participate inour research. They can come to our website and we'll offer them a free kit,"Auton said.

The contents of a 23andMe kit.(Photo: 23andMe)

"We're very much interested in trying to get the word out so that people to hear about this because really every data point is going to be pretty valuable."

23andMe has emailed customers in areas hardest hit so far in the pandemic including those in Michiganto let them know about its study, Auton said.

"The best thing that we can do to make a difference for COVID is to really publish the results that we find and make them available to the research and scientific communities," he said.

23andMe haspublishedmore than 150 studies in peer-reviewed scientific journals, "the majority of which come from collaboration with the broader academic and the scientific community," Auton said,since it launched in 2006with its direct-to-consumer DNA kit.

But the company ran afoul of the U.S. Food and Drug Administration in 2013, when the agency ordered 23andMeto halt the release of genetic health information to customers, saying the company had yet to prove its tests were"analytically or clinically validated."

After revamping, the company passed FDA muster in 2017, and got authorization tooffergenetic healthreports that outlinedrisk for 10conditions, including late-onset Alzheimers disease andParkinsons disease.

Ancestry is new to the health genetics business. It launched AncestryHealth in 2019, with the disclaimer that its tests are physician-ordered and not diagnostic, but offer "health insights" into whether a person might be a carrier for cystic fibrosis or sickle cell anemia or whether there's a genetic variant associated with a higher risk for breast cancer or colon cancer.

Ball said Ancestry also will seek to publish its COVID-19 research findings, too.

"We will be doing our very best to publish our findings as quickly as possible, and making them as useful to clinicians and other researchers as quickly as possible," she said.

Ancestry DNA(Photo: Melissa Rorech)

Both companies are looking for research partners for the coronavirus studies. Ancestry has had nibbles from universities, biotech and pharmaceutical companies, but Ball said, right now, the focus is on safeguarding the privacy of its customers.

"We typically do not share data out with third parties," Ball said. "That's an unusual activity for us.

"We will not be sharingpersonal data. Everything will be de-identified. So names, email addresses, your address, your ZIP code, your phone number, all that stuff will be stripped and will not be shared. We do want to still be very conservative because it is people's genetic data."

At 23andMe, individual-level data is never shared with a third party "without explicit additional consent from participants," Auton said.

"The information that we're talking about here, where we would be working with the academic community, is all aggregated at a very high level. So it's really just information about whether a specific genetic variant is associated with the disease. It doesn't contain any information about the individuals in the study."

Ball urged people to consider participating in this research for the common good.

"The people who came to AncestryDNA were interested in finding out about their ancestors, their past and their history," she said."This is our chancein this moment of history ... to take 5-10 minutes ... and do our best to help our community of friends or familyand the people who we don't even know who will be coming along later.

"It's our chance to contribute to the benefit of everybody. And I think right now, it'san opportunity that resonates with a lot of people."

Auton said the research could lead to therapies or treatments for people sickened by COVID-19.

"Hopefully, that can make a difference," he said.

Contact Kristen Jordan Shamus: 313-222-5997 or kshamus@freepress.com. Follow her on Twitter @kristenshamus.

Read or Share this story: https://www.freep.com/story/news/health/2020/05/26/genes-dna-ancestry-23-andme-coronavirus-covid-19/5223568002/

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Genetic genealogy companies Ancestry, 23andMe begin COVID-19 research - Detroit Free Press

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NEW YORK Employing tumor-normal sequencing, researchers have demonstrated that a significant number of advanced cancer patients learned germline findings that informed the treatment they received.

In one study, involving 12,000 patients treated at Memorial Sloan Kettering Cancer Center, researchers identified nearly 600 patients with recurrent or metastatic cancer who had actionable germline mutations, and 44 percent of them received targeted drugs either as part of the standard of care or as part of a research protocol.

Presenting the findings at the American Society of Clinical Oncology's virtual annual meeting, MSK's Zsofia Stadler said her group's study represents the most comprehensive assessment of the clinical utility of germline variants for guiding targeted therapy decisions in advanced cancer patients. This study, she said, demonstrates the "the importance of germline analysis for cancer treatment."

In another study, presented at the same meeting, researchers from the University of Michigan investigated the prevalence of actionable germline mutations in a cohort of around 1,000 patients, and found that 49 patients, or close to 5 percent, had therapeutically targetable germline mutations.

The findings from these two studies are timely given that cancer patients are increasingly having their tumors sequenced in the hopes of identifying precision therapy options. Studies have shown that after tumor profiling, approximately 10 percent of patients received results based on which they can receive precision drugs. Patients' tumors are often profiled using next-generation sequencing (NGS) panels that gauge hundreds of genes and also pick up clinically significant germline mutations, but often these findings are censored and not reported back to or discussed with the patient. This is likely because there is a perception that germline genetic mutations, which have been historically important for inherited cancer risk assessment, don't really impact the immediate care of patients.

"We know the identification of germline alterations has important implications for our cancer patients, including implementation of appropriate cancer surveillance measures, potential risk-reducing measures, and of course predictive genetic testing for at-risk relatives," said Stadler. "However, less is known about the clinical impact of germline findings on targeted cancer treatment."

At MSK, cancer patients can have their tumors profiled with the MSK-IMPACT NGS panel which gauges more than 400 genes. The goal of this testing, Stadler said, is to identify genetic mutations in the tumor that can be targeted with treatments. To do this, the cancer center sequences a patient's tumor tissue and normal blood sample, compares the detected mutations, and subtracts out the germline variants that occur in every cell in the body, not just the tumor. At the end of this process, only the somatic mutations found in the tumor are reported.

Starting in 2015, patients at MSK who received testing with MSK-IMPACT could provide consent under an institutional review board-approved protocol to receive separate germline testing for 88 genes associated with increased cancer risk. Between 2015 and 2019, nearly 12,000 patients agreed to this additional testing to learn if they harbored likely pathogenic or pathogenic mutations in any of these genes. If patients had pathogenic findings, this was noted in their medical record, which investigators then reviewed to assess whether the mutations could inform treatment.

Around half of these 12,000 tested patients had breast, prostate, pancreatic, or colorectal cancers, while the rest of the patients had a variety of rarer tumor types. Approximately 2,000 patients, or 17 percent, had likely pathogenic or pathogenic mutations, 682 of whom had mutations in high- or moderate-penetrance genes.

In terms of therapeutic actionability, Stadler and colleagues identified targetable mutations in 849 patients, or 7 percent. Since PARP inhibitors now can be given to patients with BRCA1 or BRCA2 mutations with certain types of cancers, mutations in these genes comprised more than half of the actionable findings. Nearly 20 percent of mutations were in Lynch syndrome genes, which can guide immunotherapy use.

MSK classifies the actionability of somatic variants detected by MSK-IMPACT using a three-tier system that emerged in the process of garnering FDA authorization for the panel. In authorizing that test, the FDA released a three-tier framework to help labs determine what information they could accurately communicate in test reports based on the evidence underlying detected biomarkers. The same system can also help oncologists prioritize which findings are most informative for their patient's care from the long list of genetic mutations often identified by these tests.

In the FDA's framework, tier 1 biomarkers are those that the agency has given companion diagnostic status based on evidence showing that they can determine which patients will or will not respond to a drug. Tier 2 biomarkers are "cancer mutations with evidence of clinical significance" that doctors can use in the care of cancer patients in line with guidelines and other information. Tier 3 biomarkers are "cancer mutations with potential clinical significance," which can help direct patients to clinical trials.

Since germline variations don't have a widely accepted system of classification for therapeutic actionability, Stadler and colleagues adapted this three-tier system for somatic variants to weigh the evidence on the 88 cancer risk genes. While 849 patients, or 7 percent, had targetable germline mutations in tier 1 and 2 genes, using all three tiers, around 1,000 patients, or nearly 9 percent, had germline mutations with therapeutic significance.

The researchers used the more stringent criteria germline mutations in tier 1 and 2 genes to try to guide treatment decisions for nearly 600 patients with recurrent or metastatic cancer. Ultimately, 44 percent received targeted drugs either as part of the standard of care or as part of a research protocol. Patients who received treatment had germline mutations in a variety of genes, though alterations in BRCA1 and BRCA2 drove a lot of the therapeutic decisions in the study.

As such, researchers explored this subset of patients in more detail. Of the 175 patients with BRCA1/2 germline mutations, 57 percent were classified as tier 1 mutations because patients had breast or ovarian cancer and were able to receive PARP inhibitors that had been FDA approved for their molecularly defined indication. More than 40 percent including 21 percent of pancreatic cancer patients and 11 percent of prostate cancer patients received PARP inhibitors under a research protocol. Since this study, however, the FDA has approved PARP inhibitors for BRCA-mutated pancreatic and prostate cancer.

"With the emergence of novel targeted treatments with new FDA indications, the therapeutic actionability of germline variants is likely to increase over time," Stadler said.

The study led by Erin Cobain from the University of Michigan similarly demonstrated the clinical utility of germline findings from tumor testing, though on a smaller scale. The researchers conducted targeted exome sequencing for 1,700 genes and transcriptome sequencing on tumor and normal samples from approximately 1,000 patients with advanced solid tumors. Pathogenic germline variants were identified in 160 patients, or 16 percent. Of the deleterious cancer risk mutations identified during this process, 92 percent were not known before patients were tested as part of this protocol, "which indicates that current clinical criteria may not identify all patients at risk for cancer predisposition," Cobain said.

Close to 5 percent of patients had biomarkers that could inform their treatment, such as defects in DNA repair genes, which can be treated with PARP inhibitors, and mutations in mismatch repair gene defects that can be used to prescribe immunotherapy. Significant proportions of patients with rare tumors had pathogenic variants, such as 20 percent of sarcomas, 17 percent of hepatobiliary cancers, and 16 percent of cancers of unknown primary. Cobain highlighted that some of these patients with these rare tumors had germline mutations that could potentially be therapeutically targeted, which is important given the limited treatment options in these settings.

Ultimately, 11 patients received PARP inhibitors and immunotherapies based on germline findings. Two patients had a complete response, one had a partial response, and five had stable disease. Of the three patients who had progressive disease, two were breast cancer patients with germline ATM mutations treated with PARP inhibitors, and the third was a pancreatic cancer patient with a BRCA1 mutation who received a PARP inhibitor.

The results of this study "support consideration of directed germline testing in all patients with metastatic solid tumors to identify defects in DNA repair with therapeutic targets," Cobain said.

To Funda Meric-Bernstam from MD Anderson Cancer Center, both of these studies demonstrate that if cancer centers are doing tumor/normal sequencing and looking for germline variants of clinical significance, they may not have to look too hard. However, she clarified that its also not "easy" to identify and report germline variants in this context. "This requires a lab to both analyze the normal [samples] and call pathogenic and likely pathogenic variants," she said.

Currently, there isn't a standardized method among tumor sequencing labs when it comes to dealing with germline findings. Some labs conduct only tumor testing, others conduct tumor-normal analysis but subtract germline findings, and still others perform tumor-normal testing and report only a subset of germline findings. Even if a lab sequencing only tumor tissue, tests will identify mutations in genes that are more likely to have occurred in the germline.

Meric-Bernstam noted that although the studies at MSK and the University of Michigan were done under IRB-approved protocols, in the real world when tumor sequencing is done, most patients aren't asked if they want to learn germline findings. "You really need infrastructure to return [results], to counsel, and to offer cascade testing," she said. "You need to facilitate the analysis of normal [samples] so you can ensure findings can be acted upon in a timely fashion, especially in patients with advanced disease. You need to have infrastructure in place for treatment matching and treatments available."

Though there are technical and infrastructural challenges to reporting out germline findings in the context of cancer tumor sequencing, genetics experts are increasingly of the opinion that it is no longer ethical to mask these findings, particularly given their increasingly important role in therapy selection as demonstrated by the ASCO studies. The American College of Medical Genetics and Genomics last month published a set of points that clinicians and genomics labs should consider when presumed germline variants are flagged during tumor sequencing.

Recognizing the varying practices in the field in this regard, the ACMG in its publication told genetic testing labs to be transparent about their ability to detect germline variants from tumor testing, as well as their reporting practices. Additionally, the ACMG told clinicians to take the opportunity when ordering tumor testing to evaluate the patient for clinical signs of an underlying hereditary cancer syndrome that may require germline testing.

According to Douglas Stewart, who is lead author of the ACMG paper and is a senior investigator within the National Cancer Institutes' Division of Cancer Epidemiology and Genetics, Clinical Genetics Branch, the aim of the association is to promote best practices. But it is also the association's hope, he recently said in an interview, that the issues laid out in the paper will start a discussion in the field about how "to capitalize on this huge opportunity of identifying germline variation in tumor sequencing so that it benefits as many people as possible."

Link:
Germline Results From Tumor-Normal Sequencing Guides Precision Therapy in Advanced Cancer Patients - Precision Oncology News

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Prenatal and New-born Genetic Testing Market will exceed USD 7 billion by 2024; as per a new research report.

Technological advancement and several benefits associated with infant genetic testing will be major driver of the prenatal and new-born genetic testing market. Introduction of prenatal testing has led to substantial amount of increase in adoption rate of new testing technologies such as non-invasive prenatal testing (NIPT) for detection of sub chromosomal abnormalities, single-gene disorders and aneuploidy in North America region. Inclination towards minimally invasive infant genetic testing along with demand for early detection of birth defects will one of the major reason for market growth.

Rising number of consanguineous relations in the developing countries of Asia and Middle East will foster growth opportunities for prenatal and newborn genetic testing market. The consanguineous relations are responsible for births of infants with defects and chromosomal abnormalities. Consequent, the increase in cases of live births diagnosed with birth defects will directly impact the growth of prenatal and new-born genetic testing market.

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Increasing prevalence of birth defects among the European population is one of the major reason for rising infant mortality. The necessary government initiatives in order to conduct prenatal and new-born genetic tests to diagnose the birth defects will increase the number of infant screening programs giving rise to market growth opportunities.

However, lack of infrastructure and skilled labor in the low and middle income countries will impede the parental and newborn genetic testing market growth. Moreover, ethical issues associated with prenatal and newborn testing coupled with incidences of false test results will further hamper industry growth.

Screening technology holds majority of market share in the year 2017 owing to increasing number of screening procedures of pregnant women while delivery. The rise in awareness among the families regarding genetic birth defects and early detection of genetic disorder will augment the segment growth in the near future.

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Downs Syndrome recorded largest market share in the prenatal and newborn genetic testing market due to increasing in number of births diagnosed with Downs syndrome. The rise in incidences of women getting married at later stage of life is leading to increasing births with Downs Syndrome. These factors will further support the segment growth over the forecast timeframe.

Hospitals hold the majority of market share owing to increasing number of births in hospitals. Hospitals are aided by the government and provide the facilities require for the newborn screening. The increase in number of screening programs in hospitals provides significant scope for the market growth.

U.S. dominated the market in 2017 owing to ongoing technological advancements along with favorable reimbursement policies. The U.S. region had 99% of newborn screening rate and high awareness about prenatal and new-born genetic testing will be responsible for the market growth in U.S. Presence of technologically advanced and equipped manufacturing companies along with government initiatives supporting screening activities that will impel market growth.

Some of the major market players competing in global prenatal and newborn genetic testing market are Ariosa Diagnostics (Roche), Berry Genomics, BGI, Biorad, Illumina, Laboratory Corporation of America, Natera, Qiagen, Sequenom, Trivitron Healthcare and Verinata health. The competitors are using launch of novel products with technological advancements, acquisitions and mergers as the key strategies to foresee a lucrative growth in prenatal and newborn genetic testing market. For example, In March 2018, Natera declared a partnership with QIAGEN to develop cell-free DNA assays for QIAGENs GeneReader NGS System. These DNA assays will be developed and designed for tests, including prenatal screening, for laboratories and hospitals globally.

Originally posted here:
Prenatal and New-born Genetic Testing Market Inclinations And Development Status Highlighted During Forecast Period 2019-2025 - Cole of Duty

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Womens relationship with their breasts is complicated. It includes apprehension about development, insecurity about size and shape, uncertainty about how to enhance/reveal/cover up/support them and then, post-middle age, concern about how well theyre holding up.

But nothing compares to the alarm felt when detecting a suspicious lump while taking a shower, the anxiety while awaiting the results of a mammogram or the heart-stopping terror triggered by the words Its cancer.

No one knows this more intimately than the medical professionals who interact with patients in the rapidly evolving field of breast health: geneticists, genetic counselors, radiologists, oncologists and nurse navigators.

Most breast cancers require a combination of different treatments, and the order and combination of those things is a whole lot more complicated today than ever before, says Dr. Blair Harkness, a gynecological oncologist at Hope Womens Cancer Centers, an arm of Mission Health. That is not a negative: We know so much more today about the different subtypes of breast cancer. Treatment is much more individualized, based on the biology of the different types of breast cancer.

You have to identify exactly what youre dealing with first, what kind of cancer, and then you can put together the plan and order of things. Its an important multidisciplinary team: genetics, surgery, medical oncology, radiation oncology, radiology, pathology. We all work together.

Dr. Jennifer McAlister, a breast surgeon at Pardee UNC Health Care, concurs. Each patient at Pardee receives multidisciplinary care, she explains. The entire breast cancer team is on one floor in the cancer center, so its easy for us to talk about specific cases. We also have a multidisciplinary tumor board that meets weekly to discuss all cancer cases. That team includes a radiologist, pathologist, medical oncologists, radiation oncologist, surgeons and palliative care.

Meanwhile, the current pandemic has further complicated efforts to diagnose and treat the disease (see sidebar, Breast cancer in the time of COVID).

Physicians encourage adult women of all ages to perform monthly breast self-exams. At age 40, women are advised to get a first baseline mammogram. Mission recommends an annual screening beginning at 40, because that reduces your risk of dying from breast cancer by 40%, says Dr. Sheri Fleeman, assistant medical director of breast imaging at Mission Health. One out of 6 breast cancers are diagnosed in the 40-to-49-year-old age range. Our goal is to find cancers early, treat them early and save as many lives as possible, and we feel like we can do that best if we start at 40 and do an annual.

There are exceptions, however. We recommend a baseline mammogram at age 40 unless a patient had a direct family member previously diagnosed with breast cancer, said Linda Richards, who has since retired as administrative director of cancer services at AdventHealth Hendersonville (formerly Park Ridge Health). For example, if a womans mother was diagnosed with breast cancer at 42, the recommendation for baseline for this woman would be 32: 10 years prior to the family members diagnosis.

Documenting family medical history is now standard procedure: For their first appointment at any practice, all new patients complete a lengthy questionnaire. When that information raises a red flag, patients may be referred to a genetic counselor.

Providers are becoming a lot more proactive asking about family history, and if there is cause, they may suggest an appointment at a genetics center, says Mission Healths Carolyn Wilson. We see people here for all types of indications, but the biggest area of growth, here and at all genetics centers, is in the area of inherited cancer, notes Wilson, one of six genetic counselors at the regions only such facility.

We know that about 10% of cancer is genetic, so its very common for people who either have a family history of cancer or a new diagnosis of breast cancer or another type of cancer to be referred to genetics. Determining if they might be in the genetic group, she continues, can certainly impact their screening, their treatment and medication choices.

Patients can expect the first appointment with a genetic counselor to run from 75-90 minutes. In addition to reviewing both personal and family cancer history, counselors will discuss the benefits and limitations of genetic testing. The process, says Wilson, is kind of complicated, and the results are not always black or white. We sometimes find an uncertain genetic change, which is not the same as a positive. You might not get a yes or no answer.

For those who decide to go ahead with it, the process involves either a blood or a saliva test. It takes two to three weeks to get the results, And if anything comes back positive or uncertain, we are happy to meet again to review the information, says Wilson. The report and a detailed letter are sent to them and their provider, so together they can map their next steps.

Even for those who test negative, however, the center creates a risk assessment based on personal and family history. Nongenetic factors that increase breast cancer risk include increased breast density shown on mammograms, having ones first period at a young age, having children later in life or not at all and hormone replacement therapy.

If a persons lifetime assessment risk for breast cancer is greater than 20% based on these computer-based models, they are recommended to follow high-risk breast screening, which includes an annual mammogram and breast MRI, notes Wilson.

And while mammograms are still the procedure most laywomen think of when it comes to getting naked and having your breasts squeezed flat between plates, breast imaging is a more accurate description of the many tools radiologists currently use to see the breast.

The 3D mammogram, explains Dr. David Onofrey of AdventHealths Medical Group Breast Center, creates a better image by pulling multiple X-ray images in just 10 seconds. A computer puts the images together, and this produces concisely focused three-dimensional images throughout the breast. Studies show that 3D mammography detects slightly more breast cancers than standard. AdventHealth, Mission and Pardee all perform this procedure.

At Pardee, notes McAlister, all mammograms are now 3D. In patients with dense breast tissue, we know that 3D mammograms find smaller masses or cancers that could be hidden on traditional 2D mammograms, she says. When the breast tissue is dense, it tends to overlap on itself and can be difficult to read. The 3D imaging allows us to view the breast in slices, so there is less risk of missing something small.

Another useful technology for women who have dense breast tissue a condition that, when discovered in a standard mammogram, must be reported to both physician and patient is the 3D whole breast ultrasound, or ABUS. The procedure, says Fleeman of Mission Health, uses sound waves to take 3D pictures of the breast. This is helpful because the breast is composed of fatty tissue and glandular tissue. The more glandular tissue you have, the denser your breasts. On mammograms, the fatty tissue is black and the glandular tissue is white. Cancers also show up as white, so in dense breasts, its like looking for a polar bear in a snowstorm. ABUS is a totally different way of looking at breast tissue, and in patients with dense breasts it can pick up cancers a mammogram might miss.

Yet another advanced tool is a breast MRI, which requires an IV to provide contrast. A more expensive test, its reserved for high-risk patients, says Fleeman.

If a tumor is detected, the next step would be a biopsy, and here too, advanced technology plays a role. The 3D-guided biopsy provides the radiologist with a clearer, more detailed image, allowing precise, accurate sampling of the tissue, explains Lisa Gundersen, clinical operations director of cancer services at AdventHealth. Its also more comfortable for the patient, she continues. AdventHealth, Mission and Pardee all offer this procedure.

For patients diagnosed with cancer, the interdisciplinary team develops an individualized treatment plan. Its not enough to simply say breast cancer anymore, notes Harkness. Its What kind? Because treatments we do are far more individualized for both the patient and the type of breast cancer were dealing with. So the first step is really understanding the imaging, the biopsy result and the subtype of cancer someone has, and then coming up with the best order and combination of treatment.

Most breast cancers, he continues, need some combination of surgery, radiation, chemotherapy and hormonal therapy; some cancers need all, but not every one. The order and combination depend on the subtype and the stage.

With new tests, says Harkness, oncologists can ascertain the tumors genetic makeup and, based on that, determine whether chemotherapy is likely to be effective. We use a lot less chemo for breast cancer than we used to, but hopefully were giving it more to people who will actually benefit from it. He adds that while some chemo regimens are harder to tolerate, thanks to better anti-nausea medications, the experience isnt nearly as bad as it used to be.

Despite the challenges, professionals engaged in the battle against breast cancer find reasons for optimism.

For Fleeman, the recent advances in sophisticated imaging are a key factor. We like 3D because we see better and its harder for the cancers to hide, she says. Our goal is to find and treat cancers early. With screening-detected cancer, the patient has to have less extensive surgery and less chemotherapy. When we find a cancer at Stage 1, its just a bump in the road.

Wilson, the genetic counselor, cites insurance companies increased acceptance of genetic testing. They used to look at it as kind of experimental, but now its really standard of care, because it can really make a difference financially and medically. If, through genetic testing, you can get someone the right treatment from the beginning versus failing five other treatments first, it makes a tremendous difference and impacts outcomes.

Harkness, meanwhile, believes, The biggest thing with breast cancer is how much research is being done. We have seen such dramatic strides in different areas of breast cancer over the last couple of decades; we are constantly refining and making advances. While nobody can know what that next thing is going to be, the trajectory of how much better we do in finding, treating and curing breast cancer today than we did even 10-15 years ago is where I find optimism.

Link:
Recent advances are transforming breast cancer treatment - Mountain Xpress

Recommendation and review posted by Bethany Smith

A report, added to the extensive database of verified Market Research titled Direct-To-Consumer (DTC) Genetic Testing Market 2020 by Manufacturer, Region, Type and Application, Forecast up to 2026, is intended to highlight first-hand documentation of all the best implementations in the industry. The report contains an in-depth analysis of current and future market trends, segmentation, industrial opportunities and the future market scenario, taking into account the forecast years 2020 to 2026. It contains extremely important details on the key players in the Direct-To-Consumer (DTC) Genetic Testing market as well as growth-oriented practices, that they normally use. The report examines a number of growth drivers and limiting factors. The key forecast information by region, type and application with sales and revenue from 2020 to 2026 is included in this report.

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Top 10 Companies in the Direct-To-Consumer (DTC) Genetic Testing Market Research Report:

Competitive landscape:

The report examines the major players, including the profiles of the major players in the market with a significant global and / or regional presence, combined with their information such as related companies, downstream buyers, upstream suppliers, market position, historical background and top competitors based on the Sales with sales contact information.

Regional Description:

The Direct-To-Consumer (DTC) Genetic Testing market was analyzed and a proper survey of the market was carried out based on all regions of the world. The regions listed in the report include: North America (United States, Canada, and Mexico), Europe (Germany, France, United Kingdom, Russia, and Italy), Asia-Pacific (China, Japan, Korea, India, and Southeast Asia), South America (Brazil, Argentina , Colombia etc.), Middle East and Africa (Saudi Arabia, United Arab Emirates, Egypt, Nigeria and South Africa). All these regions have been studied in detail and the prevailing trends and different possibilities are also mentioned in the market report.

Sales and sales broken down by application:

Sales and sales divided by type:

In addition, the report categorizes product type and end uses as dynamic market segments that directly impact the growth potential and roadmap of the target market. The report highlights the core developments that are common to all regional hubs and their subsequent impact on the holistic growth path of the Direct-To-Consumer (DTC) Genetic Testing market worldwide. Other valuable aspects of the report are the market development history, various marketing channels, supplier analysis, potential buyers and the analysis of the markets industrial chain.

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Table of Content

1 Introduction of Direct-To-Consumer (DTC) Genetic Testing Market

1.1 Overview of the Market1.2 Scope of Report1.3 Assumptions

2 Executive Summary

3 Research Methodology of Verified Market Research

3.1 Data Mining3.2 Validation3.3 Primary Interviews3.4 List of Data Sources

4 Direct-To-Consumer (DTC) Genetic Testing Market Outlook

4.1 Overview4.2 Market Dynamics4.2.1 Drivers4.2.2 Restraints4.2.3 Opportunities4.3 Porters Five Force Model4.4 Value Chain Analysis

5 Direct-To-Consumer (DTC) Genetic Testing Market, By Deployment Model

5.1 Overview

6 Direct-To-Consumer (DTC) Genetic Testing Market, By Solution

6.1 Overview

7 Direct-To-Consumer (DTC) Genetic Testing Market, By Vertical

7.1 Overview

8 Direct-To-Consumer (DTC) Genetic Testing Market, By Geography

8.1 Overview8.2 North America8.2.1 U.S.8.2.2 Canada8.2.3 Mexico8.3 Europe8.3.1 Germany8.3.2 U.K.8.3.3 France8.3.4 Rest of Europe8.4 Asia Pacific8.4.1 China8.4.2 Japan8.4.3 India8.4.4 Rest of Asia Pacific8.5 Rest of the World8.5.1 Latin America8.5.2 Middle East

9 Direct-To-Consumer (DTC) Genetic Testing Market Competitive Landscape

9.1 Overview9.2 Company Market Ranking9.3 Key Development Strategies

10 Company Profiles

10.1.1 Overview10.1.2 Financial Performance10.1.3 Product Outlook10.1.4 Key Developments

11 Appendix

11.1 Related Research

Customized Research Report Using Corporate Email Id @https://www.verifiedmarketresearch.com/product/Direct-To-Consumer-DTC-Genetic-Testing-Market/?utm_source=WCS&utm_medium=001

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Impact of Covid-19 Outbreak on Direct-To-Consumer (DTC) Genetic Testing Market 2020 Trends, Growth Opportunities, Demand, Application, Top Companies...

Recommendation and review posted by Bethany Smith

Since he was born, Tylen has known he had a half brother on the other side of the country. In 2008, Tylens mother, Christy Coyle, had used the same sperm donor as the other mom, Lauran, and they found one another online. The kids were born weeks apart, met as infants, and played together. Their single mothers spoke weekly on the phone and raised them as half siblings so they would always have someone just like them whom they could turn to. For years, a large canvas photo print of Coyle holding the two boys hung on a wall in Laurans house.

We had planned that when they turned 18 and got to meet the donor that they were going to do it together, Coyle said. It was just our life plan.

With their children approaching 10, the two women paid for an AncestryDNA test to provide the boys with a piece of paper confirming their ties and breaking down their genetic history. One evening, while on the phone with Coyle, Lauran looked up the results.

And then it was just dead silence, Coyle said. There was no noise at all. And she just said, Theyre not related.

Coyle, now 41, was given the wrong sperm sample. The discovery sent her into a spiral. She felt violated, having been inseminated by a strange man whom she did not choose. She felt foolish, having searched in vain for physical similarities between the two boys. But she also felt that she had failed as a mother, as if she had betrayed her son and robbed him of the life plan she had envisioned.

It was a couple of months of just sitting there, being devastated for my son, she told BuzzFeed News. Lots of no sleeping or crying because you dont know what to tell your son. You feel like youve been lying to them about where they came from.

It would take months and a DNA test from an unsuspecting middle-aged woman living far away in Texas before the mystery would be solved.

Tylen, right, with Lauran's son as toddlers.

In the multibillion-dollar sperm bank industry, stories of mix-ups have become increasingly common. While federal regulators require that samples be tested for communicable diseases such as HIV, there is little to no national regulation beyond that: No laws punishing sloppy record-keeping at the clinics, no laws mandating that the personal information provided by donors is verified, no laws ensuring that women are being inseminated with the exact samples they have selected. And theres currently no major lobbying effort to change any of that.

Wendy Kramer cofounder of the Donor Sibling Registry, a website that connects children who were conceived using sperm donors said most people dont know how little the industry is regulated. I think for so many of us when we had to use a donor in order to have a child we all thought, Oh, its the medical industry. These are medical professionals, so theres going to be the same ethics and morals and responsibilities and record-keeping, she said. What many of us have come to realize over the years is this isnt the medical profession. These are sperm-sellers, and thats very different, and their ethics and responsibilities are very different.

The increasing ubiquity and availability of at-home DNA testing kits, genealogy websites, and social media has been a slow-building storm for the donor industry, gradually exposing more and more cases of samples or records being mishandled. Just as police departments are cracking decades-old cold cases using genetic testing, donor-conceived children are learning shocking information about themselves and their families.

The mistakes from clinics span the country and stretch back decades, taking a profound human toll not only on children such as Tylen but also on donor-conceived adults, shattering their understanding of themselves and their families as well as the bonds they have painstakingly built.

We obviously didnt grow up with one another, and we obviously didnt live close to one another, so that already makes it difficult to maintain a relationship, said Sam Johnson, a 29-year-old New Yorker who unearthed shocking information about his donor and his supposed half siblings after taking a DNA test. But then when you add the fact that were not even related, its like, whats really holding it together?

These stories from Coyle and Johnson touch on fundamentally human questions: What is family? And what is its purpose?

For most people, of course, family goes beyond simple blood bonds but involves social connections built over years of shared experiences. Its also a central means by which we get to know ourselves and build our identity.

For donor-conceived children, discovering new relatives with similar lived experiences can mean theyre extending their family in both the genealogical and social sense. The half sibling could fill in gaps in their biological identity and build a relationship that fills in part of their social identity and self-awareness.

Its hard enough for these donor-conceived people to kind of jockey around the whole idea of identity: Who am I when I dont know where half of me comes from? Kramer said. Connecting with others who also share that unknown half of yourself can be very important, can be profound and life-changing. Relationships are made, bonds are made, and friendships are made. Their family has expanded.

Then to have the rug pulled out under you, to say those people are not actually your biological relative, is extremely upsetting.

Johnson knew at an early age that his family was different. Born to two moms in the early 1990s, he grew up an only child in northern Manhattan. Liberal New York was something of a bubble, but kids were still kids. I definitely got shit ... Every time somebody would say, Oh, thats gay, I would fucking correct that, he said. People would definitely make fun of the way that I was born, the fact that I was conceived through artificial insemination.

He loved his moms, but there was an occasional nagging feeling a sense of mourning, as he described it, for not knowing exactly where he had come from. The only clue Johnson had was the donor information card from the New York clinic, Repro Lab, that his birth mother, Nicole Johnson, had used to conceive him. According to the donor sheet, Donor #19 was Italian, Catholic, and worked as a doctor. He had green eyes and dark brown hair. He was married and enjoyed soccer and antiques. He didnt smoke or drink, and he had a spotless medical history. Describes himself as, read the sheet, optimistic, exciting, and honest.

As he grew older, Johnson internalized what little information the paper provided him. He tried picking up some Italian using Duolingo and even had a friend teach him some recipes, soon perfecting the simple Neapolitan dish spaghetti aglio e olio. Occasionally, hed search online for doctors photos, wondering if he looked like any of the men in the results. In 2007, his googling led him to Kramers website, the Donor Sibling Registry.

Within a year, he had connected with a woman who had the same Donor #19 card from Repro Lab. And then another. And another. And another still. He was overwhelmed.

One of the women, Genna Ellis, was the same age as Johnson; she had grown up in Brooklyn, also with two moms. When she and Johnson met at Manhattans Union Square, what could have been an awkward encounter quickly eased into an hours-long walk as they discussed their similar childhoods. It was a powerful experience for both of us, considering that was the first time we thought we were communicating with someone we were blood-related to, Ellis said.

For the next decade, Johnson reveled in these newfound connections: A social worker by trade, he helped Ellis with an addiction problem. He visited one half sister in California when he helped a friend move across the country. He helped a third through a bad breakup and let her sleep on his couch. He was there when she got her first tattoo. She even came to his wedding.

I was always proud to be like, yeah, this is my sister to be able to say that, he said. I liked the idea of it.

The first crack came when one of the supposed half sisters took a 23andMe test and learned she had Baltic, not Italian, heritage. She later matched with a Slovakian man who confirmed he had been a donor at the New York clinic in his youth.

Johnson took a 23andMe test. Ellis took one too. None of them were related to each other. Their donors were all different.

It definitely hurt to find out that these relationships not to minimize their significance but to find out that the foundations that they were built upon were false, he said. Over the course of 10 years, I spent time forming relationships with these people. Not that they werent still meaningful, but its like the foundation is, like, what the fuck?

William Henry Pancoast (1835 - 1897). American physician and surgeon.

Secrecy and fraud have been present since the very first recorded case of successful artificial insemination in the US when in 1884, a 41-year-old wealthy Philadelphia business merchant and his 31-year-old wife came to see one of the citys most prominent physicians, William Pancoast. The doctor and his medical students inspected both and ultimately concluded the mans semen contained no sperm. After months of treatment failed, Pancoast came up with a plan B when one of his students joked that the only solution to this problem is to call in the hired man.

Pancoast invited the woman to his clinic and knocked her unconscious with chloroform. Then, without her consent and with his students present, the doctor used a hard rubber syringe to inject some fresh semen from the best-looking member of the class into her uterus before plugging her cervix with gauze. The students and Pancoast made a pledge of absolute secrecy.

She became pregnant, and Pancoast eventually felt guilty enough to tell her husband. He turned out to be delighted with the idea but asked that his wife not be informed. She later gave birth to a son. The insemination was only revealed 25 years later when one of the students, Addison Davis Hard, published a piece in a medical journal recounting the procedure. By all accounts, the woman was never told.

In the 1950s, scientific breakthroughs came with the freezing of sperm and the early pioneers immediately envisaged grand commercial potential. One of the first researchers in the field, Raymond Bunge, predicted in a letter to his mother, It wont be long before my icicles will be in the deep freeze section of supermarkets. But the research remained controversial; a 1954 headline about three babies born through Bunges method of freezing and insemination declared, Fatherhood After Death Has Now Been Proved Possible. Fear of so-called test tube babies abounded.

The first sperm banks didnt arrive until the 1970s, mainly as a place for men to deposit and store their own samples for later use, say, if they were going through cancer treatment. Fertility doctors were still mostly using fresh sperm samples not frozen ones to inseminate women. When the AIDS crisis began and several women contracted HIV from fresh donations, medical preference shifted for the first time to using frozen samples for the procedure. The only real regulation implemented involved screening the samples for STDs. But the list of diseases that are tested are relatively short, said Naomi R. Cahn, law professor at George Washington University and author of Test Tube Families: Why the Fertility Market Needs Legal Regulation. And for all we know the same donor that could be rejected at one bank is going to another bank and trying again until that donor succeeds.

There are no records of how many sperm donations are made every year in the US, nor of how many children are conceived, but it is frequently said to be between 30,000 to 60,000 births per year. Kramer with the DSR argues that number is woefully out of date.

Still, the scandals tend to find the media spotlight. There was the man whose samples were used to produce more than 150 children, the many parents who end up unintentionally having children of different races (in one instance because a clinic was said to have confused Donor 380 with Donor 330), the woman whose dead husbands samples were misplaced and allegedly used to impregnate other women, or the Georgia clinic that allegedly marketed a donor to hopeful parents as a neuroscience genius studying a PhD in engineering but who was in reality an ex-con who had never attended college and had a history of psychiatric hospitalization. His sample has been used to conceive at least 36 children.

Many of these mistakes would not have come to light had it not been for the arrival of the DNA genealogy market an industry estimated to be valued at over $3 billion in the US alone. Even if donors had requested decades ago to remain private, genealogy websites and social media have made that largely impossible. A 2018 survey of almost 500 donor-conceived people conducted by Kramers website found that almost a quarter of them had used DNA testing to track down their donor.

As more donor-conceived children find their biological parents and half siblings, new and extended family units are forming. One 2016 study of 419 donor-conceived children found more than a third of them get together once a year with the half siblings theyve discovered, and a fifth of them meet up together several times a year. Some 42% of them said they considered their half sibling to be part of their immediate nuclear family.

Tylen had been Christy Coyles miracle. In 2008, she was single and working in the records department of a police department in Chicago's suburbs when she was told she needed surgery to treat cervical cancer. If she wanted to ever have children like she had always dreamed, her doctor said, she needed to move quickly. She got to work.

Within three days, she had selected a sperm bank that would mail her a sample, NW Cryobank in Washington state, and begun printing out information sheets about possible donors, organizing them on the floor in stacks. She made lists of her preferred qualities blue eyes, blonde or brown hair, athletic, good eyesight, and above all healthy and circled donors who met her criteria.

Some nine months later, she was holding her son after he was delivered via C-section. He was just beautiful, she recalled through tears. It was like every dream that I had was in my hands, and I didnt think that was going to happen.

Using a forum on the sperm banks website, Coyle connected with Lauran, who was in South Carolina and had selected the same donor (Lauran asked that she be identified only by her first name and that her son not be named to protect their privacy). She, too, was a single woman now expecting a boy. I always thought that I would have a child in a marriage where I have somebody to share it with, said Coyle. And it was like she was somebody who, even though were just friends, she was able to understand what I was going through at the exact same time, and that was huge. It got me through a lot.

The connection deepened once the boys were born. Once they were here, we could see what they looked like and how big they were, said Lauran, now 44. We were constantly comparing: What size clothes is he in? Is he crawling yet? We went back and forth like that for years.

When the boys were almost 2, the families met. The two women booked adjoining hotel rooms in Atlanta and watched as the kids played together in parks and water fountains and fed each other fruit. Soon enough, the door between the two rooms remained open and the boys ran back and forth freely.

Coyle and Lauran's children playing together in the Atlanta hotel room

Despite living far apart, the families remained exceptionally close. Lauran and Coyle even decided to use another matching donor for their second children to further connect their families.

But when Lauran logged on to AncestryDNA during the phone call with Coyle one evening in 2018, they finally discovered the truth. A subsequent DNA test performed by California Cryobank which purchased NW Cryobanks assets in 2016 from another company, Cryo confirmed that only Lauran had been impregnated with the sample both women had requested. Coyle felt like all the planning shed done years ago, all her best intentions, had been for naught.

I thought I was doing what was best. I picked somebody who he would get to know when he turned 18. I had a piece of paper telling me what he looked like. I had all the information, and suddenly you find out none of that was true. You dont even know who you got, she said. Its a really hard thing to process. And you have to go back and really reevaluate if you made the right decision. I question myself a lot. It made me feel horrible.

There was grief, too. The vision the women had for their sons futures a half brother to call their own, a donor whom the boys could meet together when they turned 18 had disappeared. We thought, Well, if something were to happen, if the donor decided he didnt want contact, or he turned out to be a jerk, [my son] and Ty would have each other. No matter what, theyd have each other, said Lauran. And it just completely took that away from us.

Sitting on their couch in 2011 in California, Bryce Branzell and his new fianc, Ariel, were watching a TV show when one of the characters went to a sperm bank. The show brought up an old memory for Branzell, then 23, about the time hed almost donated to a clinic. When he told Ariel, they laughed about it. I was like, good thing you didnt! she told BuzzFeed News. We joked about it like, good thing you didnt and you dont have 10 kids out there!

In 2008, Branzell had returned from basic training with the Army Reserves in Montana. Money was tight. As he scanned the classifieds for jobs he saw an ad promising $500 for every sperm donation. It was easy money. He filled out an application detailing information about his physical condition, health, and education. Within a few weeks, he was called in to provide a sample so the clinic could test his fertility and determine if he could donate.

Thats when he started having doubts. He liked the idea of getting his fertility tested, but he still wasnt sure if he was comfortable with donating for real. When he turned up at the clinic for the awkward experience of masturbating into a cup, the nagging doubts were suddenly alarm bells.

At the time, I was thinking prior to this, Yeah, its a good idea. I like the fact that I can help a family have a child, he said. But then I actually got in there, and Im thinking about the long-term things that could potentially come from this and the fact that I want to have my own kids and how awkward it would be to say, Hey, boys, girls, guess what? Youve got an older sibling that you never knew about.

Branzell handed his test sample to the male technician and then apologized. He was backing out. He said he was assured his sample would be disposed of and he had nothing to worry about.

A decade passed. Branzell deployed to Afghanistan twice, first with the Army and then the Marines. He met Ariel. They married and moved to Texas, where he became a police officer in Round Rock. Ariel began flipping houses. Together, they had two boys: Conrad, 5, and Asher, 2.

Bryce Branzell and his wife, Ariel.

Then in January 2019, Branzell received a text message from his mother. She had been building a family tree and was given an AncestryDNA kit for her birthday. Now, a woman in Illinois had sent a message seeking medical information about her own son, who was conceived using a donor linked to Branzells mom. Had he ever made a donation before?

It was one of those moments where Im like, there has to be some sort of mistake here. Theres no way something could have happened, said Branzell. And then all of a sudden the thought popped in my mind that this did happen; I did provide a test sample to the company. Could that have been it?

Branzell began pacing the room as Ariel went to the womans Facebook account and began scrolling. She saw pictures of the womans son. She was shocked. He had the same chin and ears as her husband. She found photos of the boy as a baby. He looked just like their own son.

Branzell couldnt help but agree: She pulled up a picture of Tylen, he said, and I thought, Yep, he looks just like me as a kid.

A DNA test would later confirm it: Branzell was Tylen Coyles father.

The first phone call between Branzell and Tylen was full of awkward fits and starts. Hi, Im Bryce, he recalled saying to his son months later. Yeah, Im your dad? I guess? Maybe?

You could tell he was nervous about it. He wasnt as talkative, according to Christy, as he usually is, Branzell said. It was kind of awkward to begin with. Just, how do we handle this relationship now?

For both Branzell and Coyle, there is no guidebook for families suddenly joined together or, in the case of Sam Johnson and Genna Ellis, torn apart as a result of errors in the donor industry. Their relationship their understanding of what their family is is whatever they and their children want it to be.

How Branzells sperm sample ended up being used to conceive Coyles son is now the subject of a federal lawsuit. He is suing California Cryobank and Cryo for negligence, fraud, and infliction of emotional distress, among other things. (Reached for comment, lawyers for California Cryobank referred BuzzFeed News to their motion they filed on May 13 to dismiss the case. Cryo also filed its own motion to dismiss. Both companies argue they did not inherit NW Cryobanks liabilities when they purchased its stock and are not responsible for any wrongdoing.)

The Branzells had decided to contact Coyle the day after they learned of her request. Shed merely been asking for medical information about her sons donor, and they reasoned that if they were in her situation they would want the same. (Branzell has a history of blood clots, and the couple test their boys routinely.)

When the message arrived, Coyle felt like she could breathe again for the first time in months. It was a relief that oh my gosh, I finally know that at least there is a person and hes a real person that gave that donation, she said. I know it sounds dumb, but I was finally able to put a face with my sons other genetic half.

This hunt for medical information is also the driving force for Johnson in wanting answers about his donor. Hes a father himself now; he worries for his son, Phoenix, who was born last year with gastroesophageal reflux disease, despite it not running in Johnsons nor his wifes known family. What else could be lurking? He may have another rare disease that we could get tested, said Johnson, but I have no idea what the fucking medical history is.

Johnson knows that family is about more than DNA. He has good friends he considers to be Phoenixs uncles. And the relationships he built with the women he thought were his sisters still mean something to him; hes just not sure what.

I spent most of my life always wondering what the other side of me was, he said. You sort of start to piece together an identity based on this collective understanding of you guys having the same sort of situation and were like, oh, were related. We can form our own weird sober family thing! And then as soon as you start to do that and its taken away, it makes it feel like not necessarily that your time has been wasted but...yeah, it also does.

It definitely does feel like a loss, he said. It feels weird now if I were to contact them and try to talk to them, theres this voice in my head thats like, why? Not that theyre no longer important to me. I still care about them. Feelings like that dont go away, but I envision this cruel audience in my head being like, Youre being weird reaching out to these people. Theyre not even related to you.

In 2018, after he learned he was not related to his supposed half siblings, he wrote to Repro Lab, but the company said it no longer had the appropriate records on file. We understand how you feel, a company representative wrote back to him in an email provided to BuzzFeed News. There is a lot of confusion from the findings you described and we understand the emotions it has surfaced.

The reply infuriated Johnson. Have you been through this exact same experience? he told BuzzFeed News of their response. Then dont fucking tell me you understand. Im pissed.

Awilda Grillo, director of Repro Lab, told BuzzFeed News she could not say definitively what occurred in Johnsons case as she started working at the clinic after his mother ordered the sample in 1990. I dont know if it was an error in record-keeping, she said. Were talking about 28 or 29 years ago. Donor #19 was, I think, probably one of the first donors of the Repro Lab, and back then things were different.

She also suggested that the physicians who inseminated the different women may have made errors. You cant say because you have a piece of paper that you were inseminated with that donor. The only thing that could confirm that is the record of the procedure, Grillo said. I dont want to point fingers and say theyre guilty [or] were guilty. Who knows? Who knows whos guilty? But Im saying theres so many possibilities.

The New York Times reported last year in a story Grillo said was misleading that the New York State Health Department had found poor record-keeping at the clinic. The investigation was prompted when a woman discovered her 21-year-old daughter had been conceived using a sample from Repro Lab she had not originally selected.

Grillo said Sams story was unfortunate and upsetting, but she had no answers for him. Theres no clarity, she said. Theres a lot of unknowns, unfortunately.

In three states California, Indiana, and Texas so-called fertility fraud laws now make it a crime for a doctor to knowingly inseminate a patient with a donor whom they have not selected and given consent. These laws mainly came in response to doctors who used their own samples to impregnate unknowing women. (One Indiana doctor parented more than 60 biological children through this fraud.) But courts have been loath to find that families have suffered as a result of an accident or negligence as long as the child is healthy. The Utah Supreme Court called this the supposition that the road not taken would have led to a better result and described it a common human fallacy.

This was the judgment Coyle said she feared receiving in sharing her story with BuzzFeed News. It seems like the moms in these kinds of stories usually get responses like, You should be happy that you just have a healthy kid, she said. But they dont understand the emotional strain that it puts on you, and I think that they really need to change regulations and be held responsible for what they did.

Lauran, too, is sympathetic to her friends plight. Ive read stories of other moms. The judge kind of says, Youve got a healthy kid. What do you care? Well, because this is a person, and Ive been telling him one thing for 10 years and now Ive been lying unknowingly, she said. Hes a person. Hes not a product.

Months after the Branzells reached out to Coyle, the couples attorneys arranged for a meeting in Los Angeles. Ariel arrived first and had the awkward experience of meeting a total stranger who had given birth to her husbands child years before she had. Its just the weirdest, she said. Youre instantly connected, but youve never met them. You dont know any of their history, but you have this really strong connection.

It was there that Coyle pulled out her phone to FaceTime her son so he could speak to his father for the first time. She had waited until it was confirmed that Branzell was his donor before telling him about the mix-up. For Coyle, sharing the painful news that Tylen was not related to Laurans son was tempered somewhat by his excitement at knowing at last who his father is. He now peppers his mother with questions about Branzells work as a police officer and time in the military cool to almost any 10-year-old and wants to know if he, too, liked science when he was a kid.

Branzell, though, is taking it slow for now, at least. He and Ariel havent told their sons yet about their new half brother. He also doesnt know how many other children he may have out there, or how many times his supposedly discarded sample may have been used. He wonders what might happen if someone else turns up at his door at age 18 asking to be part of the family.

Still, he envisages a future relationship with Tylen one day. I know that he wants to have a relationship with his father, and it wouldnt be fair for me to say I dont want that, he said. Hes a kid, and I want to give him the things that he wants because he didnt get this choice. None of us did.

In South Carolina, Lauran also had to sit her son down and break the news about Tylen, but she isnt sure he really understands or has emotionally processed what happened. I dont know that he feels the full brunt of it just yet, she said.

But, she told him, Coyle and her boys are still family even if he is no longer related by blood to them. Ten years of bonds dont vanish overnight.

In her home, she still has on the wall the canvas photo print of Coyle and the kids, taken on that Atlanta trip in the hotel pool. Tylen wears a life vest, and Laurans son has floaties on as he reaches up to the camera for his mom. Coyle is beaming as she cradles them both.

Its just a good memory, said Lauran. I still think of it as when my son met his half brother, even though I know biologically its not true. Thats how I still like to think of it.

Excerpt from:
They Grew Up Believing They Were Half Brothers From The Same Sperm Donor. A DNA Test Revealed The Truth. - BuzzFeed News

Recommendation and review posted by Bethany Smith

IIT Alumni Council Partners With Mumbai University

Image credit: Shutterstock image for representation purpose

A day after announcing the world's largest infectious diseases testing lab here to help fight the COVID-19 pandemic and other diseases, the IIT Alumni Council on May 29 said it has partnered with Mumbai University for the mega lab project.

The proposed MegaLab Mumbai, with a capacity to make as much as 1 crore tests a month, will be up and running from July with 1 lakh testing capabilities. It will reach full potential by early October, and will be the world's largest dedicated molecular diagnostic and genetic testing facility. MegaLab Mumbai will work with the National Centre for Nanosciences and Nanotechnology and the Innovation and Incubation Centre of the Mumbai University.

"The IIT Alumni Council is honoured to have the Mumbai University as its partner for enabling a world-class healthcare infrastructure in Mumbai for testing infectious diseases including the new coronavirus which has become a pandemic now.

"Under the partnership, the students, the faculty and the research infrastructure of the university will act as the research backbone of the MegaLab project, which will be based on the end-to-end Kodoy indigenous technology stack and will have adequate capacity for testing the entire population of Mumbai for infectious diseases, each month," said Ravi Sharma, the council president.

Mumbai University Vice-Chancellor Suhas Pednekar said, "Building on our strength in biotechnology and related research areas such as biosciences, we are now chartering a new path by partnering in a path breaking initiative promoted by the IIT Alumni Council to create one of the largest labs ever built for molecular diagnostic and genetic testing in Mumbai."

A global competition is underway to finalise more partners and also to identify the appropriate equipment sources, and consumable manufacturers. To enable the creation of an entire ecosystem to support the path-breaking initiative, MegaLab Mumbai will be partnering with research-oriented institutions of national importance in Mumbai and Delhi, Sharma said. The partnership with Mumbai University will ensure consistency, speed, quality and scale of MegaLab testing, he said.

The council has been on the forefront of catalysing and motivating students, professionals, corporates, startups and academicians to align their energies in the quest to find innovative, low-cost and rapidly scalable solutions for the pandemic.

The council has already developed 100 per cent indigenous kits which are in various stages of manufacturing and approvals so as to meet the entire requirement of developing countries, including India, China, Asean nations, the Middle East, Africa and Latin America.

MegaLab Mumbai is the biggest initiative of the council to date and aims to help design and establish the largest genetic testing laboratory for the new coronavirus and other infectious diseases like TB with an end-to-end capacity to carry out over 1 crore tests per month.

Ravindra Kulkarni, the pro-vice-chancellor of Mumbai University, said the varsity has an outstanding track record in the area of biological and medical sciences. "Several doctors who are at the forefront in the fight against new coronavirus have been our alumni and we take pride in their contribution," he said.

(with PTI inputs)

IIT Alumni Mumbai University VC covid pandemic

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Fighting COVID-19: IIT Alumni Council Partners With Mumbai University For Its Megalab Project - NDTV

Recommendation and review posted by Bethany Smith

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In addition, the report categorizes product type and end uses as dynamic market segments that directly impact the growth potential and roadmap of the target market. The report highlights the core developments that are common to all regional hubs and their subsequent impact on the holistic growth path of the Direct-To-Consumer (DTC) Genetic Testing market worldwide. Other valuable aspects of the report are the market development history, various marketing channels, supplier analysis, potential buyers and the analysis of the markets industrial chain.

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Table of Content

1 Introduction of Direct-To-Consumer (DTC) Genetic Testing Market

1.1 Overview of the Market1.2 Scope of Report1.3 Assumptions

2 Executive Summary

3 Research Methodology of Verified Market Research

3.1 Data Mining3.2 Validation3.3 Primary Interviews3.4 List of Data Sources

4 Direct-To-Consumer (DTC) Genetic Testing Market Outlook

4.1 Overview4.2 Market Dynamics4.2.1 Drivers4.2.2 Restraints4.2.3 Opportunities4.3 Porters Five Force Model4.4 Value Chain Analysis

5 Direct-To-Consumer (DTC) Genetic Testing Market, By Deployment Model

5.1 Overview

6 Direct-To-Consumer (DTC) Genetic Testing Market, By Solution

6.1 Overview

7 Direct-To-Consumer (DTC) Genetic Testing Market, By Vertical

7.1 Overview

8 Direct-To-Consumer (DTC) Genetic Testing Market, By Geography

8.1 Overview8.2 North America8.2.1 U.S.8.2.2 Canada8.2.3 Mexico8.3 Europe8.3.1 Germany8.3.2 U.K.8.3.3 France8.3.4 Rest of Europe8.4 Asia Pacific8.4.1 China8.4.2 Japan8.4.3 India8.4.4 Rest of Asia Pacific8.5 Rest of the World8.5.1 Latin America8.5.2 Middle East

9 Direct-To-Consumer (DTC) Genetic Testing Market Competitive Landscape

9.1 Overview9.2 Company Market Ranking9.3 Key Development Strategies

10 Company Profiles

10.1.1 Overview10.1.2 Financial Performance10.1.3 Product Outlook10.1.4 Key Developments

11 Appendix

11.1 Related Research

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Direct-To-Consumer (DTC) Genetic Testing Market Research Report 2020: Key Players, Applications, Drivers, Trends and Forecast to 2026 - WaterCloud...

Recommendation and review posted by Bethany Smith

By: Express Web Desk | New Delhi | Published: May 29, 2020 8:00:22 pm Keeping in mind the opportunity to replicate with smaller capacities at other places in India and in other developing counties, the MegaLab will comprise of three independent lines, namely 1 million tests per month (tpm), 3 million tpm and 6 million tpm.

The IIT Alumni Council Friday announced that the alumni body will be partnering with Mumbai University for the MegaLab Mumbai Initiative.

According to a press release by the alumni council, MegaLab Mumbai is the biggest Covid-19 initiative of the IIT Alumni Council, which aims to help design and establish the largest genetic testing laboratory for Covid-19 and other infectious diseases with an end-to-end capacity to carry out over 10 million tests per month.

Keeping in mind the opportunity to replicate with smaller capacities at other places in India and in other developing counties, the MegaLab will comprise of three independent lines, namely 1 million tests per month (tpm), 3 million tpm and 6 million tpm.

The release stated that a global competition is being launched to identify sources for equipment, consummable manufacturers and operating partners. The lab has also proposed to work with National Centre for Nanosciences and Nanotechnology and the Innovation and Incubation center of Mumbai University. The alumni of IITs and alumni companies will work with the students, faculty and alumni of Mumbai University.

IIT Alumni Council is honoured to have Mumbai University as its partner for enabling world class healthcare infrastructure in Mumbai for testing and treatment of infectious diseases including Covid19. The students, faculty and research infrastructure of Mumbai University will act as the research backbone of the MegaLab project . MegaLab will be based on the end-to-end Kodoy indigenous technology stack and will have adequate capacity for testing the entire population of Mumbai for infectious diseases, each month said Ravi Sharma, President of IIT Alumni Council. This partnership will ensure the consistency, speed, quality and scale of MegaLab testing, he added .

Commenting on the partnership, Professor Suhas Pednekar, the Vice Chancellor of University of Mumbai said, With a 160+ years history of excellence in academics, University of Mumbai has been a pioneer in research work across multiple disciplines. Building on its strength in biotechnology and related research areas such as biosciences, it is now chartering a new path by partnering in a path breaking initiative promoted by the IIT Alumni Council to create one of the largest labs ever built for molecular diagnostic and genetic testing in Mumbai with a capacity of 10 Million RTPCR tests per month.

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Covid-19: IIT Alumni Council, Mumbai University to partner for MegaLab Mumbai initiative - The Indian Express

Recommendation and review posted by Bethany Smith

The memorable scenes of loons in the 1981 movie On Golden Pond were my first introduction to common loons, but it would be more than 30 years before I saw one in person.

While kayaking with my husband at a Northeast Washington lake five years ago, we could hear their legendary haunting and eerie call. A large male common loon then swam into view, and my fascination with loons began.

As a nature photographer, I was drawn to these unique, beautiful and interesting birds.

The striking appearance and haunting call of the loon are legendary. Their plumage has intricate patterns of black and white on their bodies; a dark multicolored collar, a head and partially striped neck that are iridescent when washed in sunlight.

And those piercing red eyes.

Loons breed throughout most of Canada and northern United States. They migrate to salt water or large fresh water large lakes and rivers in winter, returning to the same nesting and migration sites each year. Loons are monogamous, but will seek out a new mate if their mate doesnt return in the spring. The lifespan of a healthy loon is up to 30 years.

Both male and female loons build their nest, incubate eggs, and care for young. Their tender devotion to their chicks is one of the most remarkable scenes I have witnessed in the natural world, and a trait that endears them to many. Chicks ride on their parents backs for the first couple of weeks after hatching.

In 2018, I was fortunate to witness this for the first time. The chick climbed onto the parents back using the tail like a ramp or going up under the parents wing. The little chick sat warm and safely protected, peering out from under the shelter of the wing, an image that remains forever etched in my mind.

Ive since viewed and photographed newly hatched chicks on multiple occasions, and the experience always leaves me in awe.

Their diet consists of fish, crayfish and aquatic invertebrates such as dragonfly nymphs. Loons swim along the surface of the lake with their head and necks underwater scanning for fish, shooting like a torpedo underwater after their prey. The parents feed chicks until they migrate in the fall.

While I was once focused on getting the shot, the motivation behind my work has changed over time. I began researching the creatures I was photographing to learn more about them, and in the process have come to really care about what I photograph and the challenges they face. Advocacy and teaching others about the subjects in my images are now an important part of my work.

There is often a story behind the scenes I photograph. Such is the case with loons. It turns out that seeing loons is a special thing indeed. Loons once nested in Washington, California, Idaho and Oregon, but today they are only known to be nesting in Washington. Historically, there were 50 known nesting pairs in Washington state. That number is much smaller now. The majority of nesting loons are in Northeast Washington, with just 16-20 nesting pairs there.

A loon pair has a clutch of one or two eggs per year. Few loon chicks survive to the end of summer to migrate. In 2000, Washington Department of Fish and Wildlife listed them as a sensitive and threatened species. Factors affecting their recovery are predation by other animals and birds, habitat loss, lead and mercury poisoning, and human interference.

In 1980, there were just 104 bald eagle breeding pairs in the state, but by 2005 had recovered to 840, continuing to increase since that time to the point that they have been removed from both the federal and state endangered species lists.

As predators, eagles sometimes prey on both loon chicks and adults a factor in building a sustainable loon population. Loons also ingest lead fishing tackle from broken fishing lines, resulting in lead poisoning. Regulations banning lead tackle at lakes where loons nest has helped, but non-nesting lakes that still allow lead tackle continue to affect mortality in transient or migrating loons.

Human interference is complex. Some loons are struck by boats or jet skis. At least nine waterfowl species eat fish in Eastern Washington, but there are a handful of people who object to loons eating fish, shooting them or intentionally destroying their nests.

Well-intentioned nature lovers and photographers also put them at risk by getting too close, driving loons from their nests or disrupting their nesting/chick raising in some way. Loons show distress by moving away from feeding chicks, leaving the nest, theatrical acrobatics in the water or alarm calls.

If your presence changes their behavior in any way, you are too close.

I have been fortunate to meet and learn from Dan and Ginger Poleschook, two volunteer researchers who have dedicated the last 25 years to Washington loon conservation work and advocacy, and have been instrumental in loon recovery.

Last summer, I joined them for loon banding at a remote Ferry County lake. Banding occurs late at night when all traces of light are gone a necessity for successful loon capture.

To an outsider, it probably looked like a clandestine drug operation, but it was actually a team of experts from the Forest Service and Biodiversity Research Institute working together on behalf of the loons.

Loons were captured and brought back to a staging area on shore, where the team drew blood to measure heavy metal levels, collected feathers for genetic testing and measured, weighed and banded the loons. A large male was caught first, and then later the female with a chick.

While the female was processed, Ginger held the tiny days-old chick in her hands. The team repeated this process nearly every night, covering nine lakes in Eastern Washington. They were knowledgeable and efficient, with an infectious camaraderie and passion for loon conservation.

Though we wrapped up around 3 a.m., I was energized by this unforgettable experience. It gave me a greater awareness that there is often much happening behind the scenes to preserve the wildlife many of us treasure.

Because of their longevity and place in the food chain, common loons are widely recognized as an indicator species that reflect the health of the aquatic environment, which in turn impacts human health.

Ameritrade founder Joe Ricketts Conservation Foundation made a substantial donation to support loon conservation and relocation of the common loon to former nesting areas where they have been eliminated. Ricketts believes the loons on our lakes and streams are similar to the canary in a mine.

If were polluting our lakes to the point where were killing the loons, its a wake up call to us as human beings that were causing more damage to our environment than we think we are and we cant see it except through a bird like the loon, Ricketts said.

Loons are dependent on our choices for their continued survival. As Maya Angelou once said, When we know better, we do better. As much as I love photographing loons, their survival is more important than any experience I might have or image that I take.

This has translated to changes in how I observe and photograph loons quietly observing loons from a distance to avoid disturbance, backing away if their behavior changes or they show signs of distress. I come back at another time if I see other people are already observing loons on a lake. The purchase of long lenses and cropping has helped to get close-up shots while maintaining a respectful distance.

I advocate for and educate others about loons when there is an opportunity. Weve replaced fishing gear with nonleaded tackle, and make every attempt to remove lines that have become snagged. These small things make a difference.

The loons are returning to Northeast Washington and nesting season is upon us. Understandably so, many people love loons and their unforgettable haunting call. My hope is that in knowing the loons plight, we all do what we can to protect these magnificent birds, so that we can continue to enjoy them for generations to come.

The loons just might have a bigger role in our lives than we think. A win for the loons is a win for us all.

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Looking for loons: Isolated birds widely recognized as indicator species of healthy aquatic environment - Sports and Weather Right Now

Recommendation and review posted by Bethany Smith

Dr. Bonnie Ferrara talks about how Renown Medical Group is offering virtual doctor visits for certain patients. Reno Gazette Journal

The ball has bounced to the patients court

Thats at least according to Renown CEO and President Anthony Slonim,who said the outcome of the coronavirus pandemic means consumers will have more options and choices.

And those options are coming faster than ever before in a medical world that was sometimes been slow to transform to patient needs.

We have been doing it around the edges, said Slonim about the incremental progress the medical community made on population health and consumer-driven healthcare.

Overnight our world has shifted to make us think more deliberately about how we can go about executing it.

Slonim said the expansion of telemedicine happened literallyovernight in Nevada as patients with other illnesses avoided doctors offices and urgent cares for fear of contracting coronavirus.

We are learning that people want convenience in their living rooms.

He saidtelemedicine is the just one of the ways consumers are demanding options.

He said now when a person is sick they can decide if they need to Skype a doctor.

Do they need to have a nurse practitioner visit them at home or do they need to go to an urgent care or the ER?

Slonim said medicine is changing and giving concierge service to everyone.

Slonim also said the problem in medicine had been there hasnt been a quarterback.

Now some of that ownership is back on primary care physicians.

I am telling doctors, 'Dont wait for them to come to you,' he said. You have to manage it.

Dr. Bonnie Ferrara of the Renown Medical Group poses for a portrait while using the Zoom app on her computer on May 20, 2020. This is how Dr. Ferrara would appear to patients during a virtual medical consultation.(Photo: JASON BEAN/RGJ)

He also said health care will need to learn to relinquish some of the control.

We learned that with our Healthy Nevada Project, Slonim said of an initiative pioneered by Renown to genetically test thousands of Nevadans to evaluate the health of the population.

When we made genetic testing free, 50,000 people signed up, he said.

He said the same is true of COVID-19 testing.

Everyone you talk to wants a COVID test, he said. Opening it up to everyone without a doctors prescription puts consumerism more front and center than ever before.

He said it gives families and individuals choices on how to manage their own health.

There are safe and appropriate things you can do without having to go to a doctor, he said. Needing a doctors prescription for tests is old school thinking.

And while the image of coronavirus focused on social distancing and doctors covered head to toe in protective equipment, Slonim said the pandemic has the medical community talking about the human touch.

During a recent medical webinar, Slonim said the conversation was that technology cant be everything.

Remember when it was OK to sit on the bed with a patient and hold their hand, Slonim said a doctor on the call said.They talked about how it became common to view that as invading a patients space.

But Slonim said people still long for that connectivity and caring.

What will evolve is a way to understand how to do that differently with technology enabling it.

3 ways healthcare will change for patients

Siobhan McAndrew tells stories about the people of Northern Nevada and covers education in Washoe County. Read her journalism right here. Consider supporting her work by subscribing to the Reno Gazette Journal.

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Consumer healthcare choices and options to be an outcome of COVID-19 pandemic - Reno Gazette Journal

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Ask anyone in North Dakota wildlife circles, and theyd likely be hard-pressed to name an animal that generates a bigger buzz than a mountain lion that makes its presence known.

It depends on a persons passion but mountain lions do even for the nonhunting public garner a lot of interest from people, said Stephanie Tucker, furbearer biologist for the North Dakota Game and Fish Department in Bismarck.

That definitely was the case in late March, when authorities in West Fargo shot and killed a treed mountain lion that was deemed potentially dangerous near a local park. The cat was a long way from the core of North Dakotas mountain lion breeding range in the Badlands and Little Missouri River Breaks.

Theres way more, say, deer hunters in North Dakota that are passionate about deer hunting and follow whats going on with deer, Tucker said. But if youre not a deer hunter, you dont really care about deer and might not be fascinated by them.

Whereas, theres just a lot of people, regardless if theyre hunters or not, that are fascinated by mountain lions.

Stephanie Tucker, furbearer biologist, North Dakota Game and Fish Department. (Photo/ North Dakota Game and Fish Department)

Tucker gave an update on mountain lions in North Dakota during the Game and Fish Departments recent spring advisory board meetings for hunters and anglers held online because of COVID-19 and further discussed the iconic cats this week in a telephone interview.

Bottom line, mountain lions in the core of their western North Dakota range are holding at a level thats low but steady and acceptable both to wildlife managers and landowners, Tucker says.

Were not hearing a lot of concerns from ranchers or deer hunters who dont like to share deer with mountain lions and things like that, she said. I definitely feel, from my perspective, that weve found a very comfortable situation where were between everybodys expectations, and the population is sustaining itself at a healthy level at the same time.

In his book, The Mammals of North Dakota, UND Professor Emeritus of Biology Robert Seabloom writes that mountain lions historically occurred across the Great Plains but never were common.

There were no early records of mountain lions in eastern North Dakota, and cats that historically inhabited North Dakota west of the Missouri River had disappeared by the early 20th century, the book states.

The population likely was so low that mountain lions in North Dakota were undetectable to most people, Tucker says.

They were likely extirpated or close to there, she said. They dont have high reproductive rates. A female only produces a litter every other year, and theres probably only two kittens in each litter so it just takes a while for the population to build itself.

According to The Mammals of North Dakota, mountain lions started gaining a foothold in the western part of the state in the 1950s. By 2005, the Game and Fish Department deemed the population high enough to offer an experimental hunting season.

Game and Fish manages mountain lions by dividing the state into two zones: Zone 1 in the Badlands area encompasses the core breeding range, while Zone 2 in the rest of the state is less suitable for the cats.

North Dakota Game and Fish Department

North Dakotas mountain lion population peaked in 2011 or 2012 at a range somewhere between 100 and 275, based on results from computer modeling, Tucker said. In response, Game and Fish upped the harvest quota for cats in Zone 1 to drive the number down and slow the increase.

Current models suggest a population in the range of 50 to 60 cats.

Once the population came back down, we backed off on our hunting season harvest limits a little bit, and the population has been fairly stable, with minor fluctuations up and down from year to year, since that time, Tucker said.

Hunters shot 14 mountain lions in Zone 1 during the 2019-20 season and three cats in Zone 2. Since the inaugural season in 2005, the statewide harvest has ranged from as low as four to as many as 19 during the 2017-18 season, when a record six cats were taken in Zone 2 outside the core breeding range.

The Fort Berthold Indian Reservation also offers a season but works with Game and Fish in sharing information to manage the species, Tucker says.

North Dakota Game and Fish Department

In an effort to learn more about the high number of cats taken in Zone 2 during the 2017-18 season, Game and Fish contracted with the U.S. Forest Services Rocky Mountain Research Station in Missoula, Mont., to conduct genetic testing on mountain lions killed outside their traditional range, Tucker said.

We really got to wonder, Are these mountain lions that are coming from our breeding population, or are they coming from somewhere else like the South Dakota Black Hills or one of the mountain lion populations in Montana? she said.

Results from that testing showed about 50% of the dispersing cats came from South Dakota, 25% from Montana and the remaining 25% from North Dakota.

The numbers dont account for cats that either pass through the state safely or travel through undetected.

This is just the ones that are taken by hunters or hit by an automobile or something along those lines, she said.

Mountain lions are well known for traveling long distances, Tucker said, especially young males, a trait believed to be natures way of preventing inbreeding within a population.

Mountain lions really have turned up in all corners of the state, she said.

Results from the genetic testing have helped the department build on the knowledge base it gathered on mountain lions during a GPS tracking survey that began in 2011 and wrapped up in 2017.

As part of the survey, a partnership with South Dakota State University, Game and Fish staff and SDSU graduate students trapped and fitted 14 mountain lions with GPS collars, while 22 cats received ear tags.

The collars were programmed to provide tracking information every three to six days, Tucker said. None of the cats collared in the survey permanently dispersed from the Badlands, although a couple of cats did venture off before returning, she said.

A mountain lion in western North Dakota wears a GPS tracking collar as part of a study the Game and Fish Department conducted beginning in 2011 and continuing through 2017. As part of a study with South Dakota State University, 14 mountain lions were fitted with GPS collars and another 22 with ear tags. (Photo/ North Dakota Game and Fish Department)

Results from the GPS collar survey helped the department fine-tune the modeling it uses to determine population trends for mountain lions in the state. Other factors used to determine population trends include the age of cats taken by hunters, who are required to submit the carcasses to Game and Fish, and genetic analysis.

Mountain lions dont lend themselves to a lot of our traditional survey techniques, Tucker said. We cant do a roadside survey and expect to see a mountain lion. We cant get up in an airplane and do an aerial survey and expect to see a mountain lion. Theyre nocturnal, they exist in low densities on the landscape, they have large home ranges and so we really rely on a population model to tell us what the population trends are.

In other words, its not about specific numbers.

I cant stress that enough, Tucker said. People always ask me, How many mountain lions do you think we have in North Dakota? And I say I dont know.

What is known is that mountain lions conceivably can wander through any part of North Dakota at any given time. As a ballpark estimate, Tucker says Game and Fish gets 25 to 30 reports of mountain lion sightings every year in North Dakota, of which maybe one-half to one-third can be verified.

Im always fascinated, too, she said. One of the reasons we know that mountain lions are in eastern North Dakota is because of reports from the public like trail camera photos or somebody snaps a picture of one and it was there and gone.

Tucker encourages the public to report sightings using the departments online furbearer reporting form at https://gf.nd.gov/hunting/furbearers/rare-furbearer-observation.

Dokken reports on outdoors. Call him at (701) 780-1148, (800) 477-6572 ext. 1148 or send email to bdokken@gfherald.com.

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As North Dakota wildlife goes, mountain lions top the list of species that captivate and fascinate - Jamestown Sun

Recommendation and review posted by Bethany Smith

Company analysis of Male Breast Cancer Treatment Market size which showcase a regional manufacturing status, such as volume, Male Breast Cancer Treatment market share, value and price details.

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COVID-19 Impact and Recovery Analysis on Male Breast Cancer Treatment Market investigated in the latest research - WhaTech Technology and Markets News

Recommendation and review posted by Bethany Smith

One of the buzzwords in the hemp space this year is autoflower. But what is it and why would you want to grow it? This episode answers those questions and a whole lot more with a roundtable panel discussion about autoflowering varieties of industrial hemp with Lancaster County farmer Steve Groff, Atlas Seed Co. Breeder Joe Ullman, and Atlas Seed Co. grower Ryan Power.

And heres what we cover:

The differences between autoflower and photoperiod hemp

Is cloning an option

Expected feminization rates

When does the flowering cycle start

Best time to plant

Recommended spacing

Transplanting vs. direct seeding

Optimal feeding plan

Harvesting

Expected yields

Cannabinoid percentages and more.

For more information, check out Atlas Seed (https://atlasseed.com/) and Hemp Innovators (https://www.hempinnovators.com/)

Autoflower FAQ, provided by Atlas Seed Co.

What is the difference between autoflowering genetics and normal clone or full term varieties?

In short, autoflowering varieties, otherwise known as day neutral genetics or Cannabis ruderalis, begin their flowering phase automatically, regardless of changes in light cycles; normal, full term, clonally propagated Cannabis sativa and Cannabis indica varieties flip to their flowering phase when they exposed to 12 hours of darkness.

Can you take cuttings (clones) of autoflowering varieties?

No, autoflowering cannabis does not allow for cuttings to be taken and therefore must be started from seed and be pollinated to further genetic lines.

What can I expect in terms of the feminization rate of Atlas Seed genetics?

The seeds we offer will show between 1 and 1500 to 1 and 2500 male to female ratio in their respective population. That means between 99.93% and 99.96% will female. We also offer lab test certified feminized seed on key lots to verify the quality of our feminization process.

When can I expect my auto plants to begin their flowering cycle?

Roughly speaking (again growing climate and genetically dependant), autoflowering plants finish their vegetative cycle between weeks 3-5, will continue to stack flowering sites between weeks 3-8, and will see their flowering sites bulk up, densify, and finish in weeks 8-12. For those used to full term and yet unaccustomed to autoflowering cannabis, remain calm until the end of the cycle and watch in marvel as plants continue to increase their flowering yield and cannabinoid content up until the day of harvest!

When is the best time to plant autoflowering varieties?

This is highly dependent on your local climatic conditions, but the rule of thumb is that autos prefer long, dry, sunny days. If you are going for one solid, high yielding harvest then planting as soon as summer soil temperatures stabilize is the way to go. If you are planning on 2 harvests, we generally recommend trying to squeeze in a second late one as opposed to an early harvest, as autoflowering genetics are native to Siberia and will finish more reliably in the cold than they will begin in it.

What is the most common plant spacing for autoflowering varieties?

For hemp we are recommending between 8-12k per acre (more or less 1 plant every 4 square feet), and for cannabis we are recommending between 17,500-20k acre (or about 1 every 2 square feet). For example, 1 row on a 30 bed with 12 in-row spacings will come out to roughly 17,424 plants per acre.

Can you transplant autoflowering varieties?

Absolutely, but there is a method that must be applied to ensure yields are not affected. The most common blunder is for farmers to let seedlings go until they are rootbound which is the easiest way to shock ones plants and greatly reduce their overall yield. The best results we have seen is when plants are transplanted between 7-12 days after sowing. The trick here is to use a cell tray that makes it easy to remove small plants without damaging them. Weve seen the best results with Growcoons, but there are certainly other options as well (ellepots, ihort, etc).

Can you direct seed autoflowering varieties?

Yes, but again this must be done with proper parameters in place, i.e., seasonal timing, soil type, equipment, and so on. As of yet we have not seen any yield differences between transplanted plants and direct seeded, but we are in the process of collecting massive amounts of data on this. Also, if one plans to do this and they are a beginner, than you will likely need 2-3x the seed to see the emergence you want.

What is the most optimal feeding plan you recommend for autoflowering plants?

We recommend that seasoned full term cannabis growers continue to follow their intuition and hard won skill sets, with one key difference: Normally when full term plants initiate their flowering stage, one switches immediately from vegetative, nitrogen rich fertilizer, to flowering, phosphorous rich bloom formulas. With autos, it is important to continue using vegetative, nitrogen rich recipes until week 6-7, well after they have begun their flowering phase. This is done in order to maximize the canopy as the plants continue to grow vertically and horizontally even as they are putting on flowering sites and bulking up.

After week 6-7, transition to bloom recipes to maximize flower yield and cannabinoid potential. They will put in the majority of their weight in the final 3-4 weeks. As a general rule of thumb, remember that the entire vegetative and flowering cycles of the plant are happening in a 70-80 day period. Some slow release nutrients growers may customarily use may not be appropriate for autos.

If I plant autoflowering plants right next to my full term plants, will it cause my full terms to initiate early flowering?

No, no, and decidedly no. We have heard anecdotal hearsay on this issue but based on experience and understanding the difference in the flowering mechanisms between these genetic lines (ruderalis vs. indica / sativa), we do not believe this is possible.

How long does it normally take before autoflowering plants can be harvested?

Classic autoflowering plants (as opposed to super autos which take closer to 120 days) can be harvested between 65-90 days depending on the variety and time of year. During peak summer months when the light intensity is higher, autoflowering crops finish faster. During early spring or late fall and especially in a winter greenhouse run, autoflowering plants will take 10-20 days longer to come to full maturity.

What can I expect in terms of biomass yield for each plant?

Between 2-4 ounces, depending on all of the aforementioned factors, i.e., grower skill, genetics, soil health, etc..

What kind of per acre yield can I expect?

This will differ greatly between hemp and cannabis and most markedly depends on your planting densities. For cannabis between 2-5k lbs is common. For hemp between 1-3k lbs. Is common.

How much of my crop can I use as finished, trimmable flower?

This is most probably most dependent on genetics, but anywhere between 25%-100%. Some plants will present uniformly sized buds that are all trimmable throughout the plant, and others will present a variety of different sized buds, some of which will be more appropriately allotted to smalls or else sent off for extraction. Again, your breeder should be able to answer this question based on experience.

What can I expect in terms of total cannabinoid content from autos?

This is potentially the most grower skill dependent variable of all, but most of the best auto cannabis genetics around today are at or exceeding 20% THC, which is miraculous if one considers they are taking half as long to bring to harvest. The auto hemp game is vastly different as state by state standards for hot hemp differ greatly, however the best varieties of compliant auto hemp as a population are hitting around 30:1 CBD to THC ratios while remaining compliant. Experienced breeders will know their varieties well enough to be able to recommend optimal harvest times to remain in compliance.

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What is Autoflowering Hemp and How to Grow It - Lancaster Farming

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