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News Article | May 11, 2017
Site: globenewswire.com

Poultry Probiotics Market share is poised to cross USD 1.8 billion by 2024; according to a new research report by Global Market Insights, Inc. Shift in preferences towards using NGPs (natural growth promoters) for livestock farming with regulatory sanctions on AGPs will fuel poultry probiotics market size. Growth in meat & meat products demand due to consumer inclination towards consuming ready-to-eat processed products should drive industry demand. Supporting regulatory norms led by FDA regarding GRAS approval of probiotics along with China promoting product research and product development in feed products for animal nutrition is likely to stimulate industry growth. Request for a sample of this research report @ https://www.gminsights.com/request-sample/detail/318 Germany poultry probiotics market size for layers should witness gains at above 6.5% by 2024. Strict regulatory guidelines by EU Commission regarding antibiotics ban due to bioaccumulation causing resistance to medical antibiotics in animals as well as humans thereby stimulating industry growth in this region. The industry is gaining importance owing to technological and product innovations in this sector which should contribute towards regional growth. Bacillus based poultry probiotics market size should grow significantly to surpass USD 100 million at the end of forecast period. These are widely used in feed products for broiler chicken due to its potential in decreasing cost and concentration of animal feed. Bacillus spores are biocompatible, stable and easily available thereby promoting its use in animal nutrition products. Key feedstock includes Lactobacilli, Bacillus, Streptococcus and Bifidobacterium. These are live bacteria which are hard to maintain and test which is likely to challenge industry profitability. Standardization issue and technological limitations are major limiting factors. Browse key industry insights spread across 175 pages with 157 market data tables & 23 figures & charts from this 2017 report Poultry Probiotics Market in detail along with the table of contents: U.S. poultry probiotics demand for streptococcus may exceed USD 25 million at the end of 2024. Streptococcus plays an important role as growth promoter which helps in increasing the animal weight and enhancing feed efficiency. Prominently used streptococcus strains include S. cristatus, S. thermophiles, and S. faecium. India poultry probiotics market size from broilers application should gain above 8%. Growing meat consumption due to disease outbreaks along with government initiatives to promote agribusiness and animal nutrition products will boost regional industry growth. Poultry probiotics market size from poult & chick applications may witness gains at above 6%. Chick & poults need these products to improve gut health and microflora which would improve their immune systems. Prolong exposure towards contaminated environment may cause infections in new born chickens which can be prevented by probiotics which supply complete nutrition and promote digestive metabolism thereby maintaining livestock health. Global poultry probiotics market share is competitive and moderately consolidated. DuPont, Chr. Hansen, Kemin, Novozymes, Lallemand, DSM Evonik, Novus, Biomin, Adddiseo and Prowel are some companies active in this market. Make an inquiry for purchasing this report @ https://www.gminsights.com/inquiry-before-buying/318 Eye Health Ingredients Market size was worth over USD 180 million in 2016 and is set to exceed 1.8 kilo tons by 2024. Hectic & busy life schedules along with exposure to toxic materials have an adverse effect on the eyesight of people leading to suffer from an unhealthy vision. Increasing requirement to maintain the perfect “20/20” eyesight has alleviated the demand for eye health ingredients. https://www.gminsights.com/industry-analysis/probiotics-market Animal Feed Organic Trace Minerals Market size was over USD 490 million in 2016 and will witness CAGR over 6.5% from 2017 to 2024. Upsurge in global meat & dairy production due to its increasing consumption will drive animal feed organic trace minerals market size. The industry will witness a significant gain as trace minerals assist in activating essential hormones and enzymes which are vital for metabolic processes in livestock. Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.


News Article | May 10, 2017
Site: www.prnewswire.co.uk

"The completion of this transaction and emergence of Novus as one of the few publicly-listed ENT companies marks a significant milestone," said Gregory J. Flesher, President and CEO of Novus Therapeutics, Inc.  "Our pipeline provides a compelling opportunity for value creation as we move our product candidates through development.  With the capital obtained through this transaction and continued access to the capital markets, we will be able to accelerate development of OP-01 for acute otitis externa and OP-02 for chronic and recurrent otitis media." The combined company has approximately $30 million in cash at close, excluding $7 to 8 million in estimated transaction-related expenses.  Following the completion of the transaction, together with the $7 million concurrent financing and reverse stock split, the company has approximately 7 million shares of common stock outstanding. Novus Therapeutics will operate under the leadership of Gregory J. Flesher, President and Chief Executive Officer; Christine G. Ocampo, Chief Financial and Chief Compliance Officer; and Dr. Catherine C. Turkel, Chief Development Officer.  The Board of Directors of the company is initially comprised of seven directors:  Keith A. Katkin (Chairman), Gregory J. Flesher, Gary A. Lyons, Erez Chimovits, Cheryl Cohen, John S. McBride, and Jodie P. Morrison.  The corporate headquarters is located in Irvine, California. Novus Therapeutics is a pharmaceutical company focusing on the acquisition, development, and commercialization of ear, nose, and throat (ENT) products.  The company has two technologies, each of which has the potential to be developed for multiple ENT indications.  The company's lead product is a nasally-administered, combination drug product (OP-02) intended to address the underlying cause of otitis media and Eustachian tube dysfunction (OM/ETD), conditions that affect more than 700 million people around the world every year.  Otitis media is one of the most common diseases seen by pediatricians and the most frequent reason children consume antibiotics or undergo surgery.  The company also has a foam-based drug delivery technology platform (OP-01) that can be used to deliver drugs into the ear, nose, and sinus cavities.  The company is currently developing a foam-based combination drug-product delivered to the external ear canal that is an improved treatment option for acute otitis externa ("swimmers ear").  For more information on the company, please visit novustherapeutics.com. Any statements in this press release about the company's future expectations, plans and prospects, including statements about its strategy, future operations, development of its product candidates, the review of strategic alternatives and the outcome of such review and other statements containing the words "believes," "anticipates," "plans," "expects," "may," and similar expressions, constitute forward-looking statements within the meaning of The Private Securities Litigation Reform Act of 1995. Forward-looking statements include, but are not limited to, expectations regarding the timing for the commencement and completion of our clinical trials and our ability to accelerate the development of our drug candidates.  Actual results may differ materially from those indicated by such forward-looking statements as a result of various important factors, including: the sufficiency of the company's cash resources; the ability to timely develop and manufacture clinical batches of our study drugs; the ability to obtain necessary approvals to commence additional clinical trials; whether data from early clinical trials will be indicative of the data that will be obtained from future clinical trials; whether the results of clinical trials will warrant submission for regulatory approval of any investigational product, any such submission will receive approval from the United States Food and Drug Administration or equivalent foreign regulatory agencies and, if we are able to obtain such approval for an investigational product, it will be successfully distributed and marketed.  Any forward-looking statements contained in this press release speak only as of the date hereof and not of any future date, and the company expressly disclaims any intent to update any forward-looking statements, whether as a result of new information, future events or otherwise.


News Article | May 11, 2017
Site: globenewswire.com

Poultry Probiotics Market share is poised to cross USD 1.8 billion by 2024; according to a new research report by Global Market Insights, Inc. Shift in preferences towards using NGPs (natural growth promoters) for livestock farming with regulatory sanctions on AGPs will fuel poultry probiotics market size. Growth in meat & meat products demand due to consumer inclination towards consuming ready-to-eat processed products should drive industry demand. Supporting regulatory norms led by FDA regarding GRAS approval of probiotics along with China promoting product research and product development in feed products for animal nutrition is likely to stimulate industry growth. Request for a sample of this research report @ https://www.gminsights.com/request-sample/detail/318 Germany poultry probiotics market size for layers should witness gains at above 6.5% by 2024. Strict regulatory guidelines by EU Commission regarding antibiotics ban due to bioaccumulation causing resistance to medical antibiotics in animals as well as humans thereby stimulating industry growth in this region. The industry is gaining importance owing to technological and product innovations in this sector which should contribute towards regional growth. Bacillus based poultry probiotics market size should grow significantly to surpass USD 100 million at the end of forecast period. These are widely used in feed products for broiler chicken due to its potential in decreasing cost and concentration of animal feed. Bacillus spores are biocompatible, stable and easily available thereby promoting its use in animal nutrition products. Key feedstock includes Lactobacilli, Bacillus, Streptococcus and Bifidobacterium. These are live bacteria which are hard to maintain and test which is likely to challenge industry profitability. Standardization issue and technological limitations are major limiting factors. Browse key industry insights spread across 175 pages with 157 market data tables & 23 figures & charts from this 2017 report Poultry Probiotics Market in detail along with the table of contents: U.S. poultry probiotics demand for streptococcus may exceed USD 25 million at the end of 2024. Streptococcus plays an important role as growth promoter which helps in increasing the animal weight and enhancing feed efficiency. Prominently used streptococcus strains include S. cristatus, S. thermophiles, and S. faecium. India poultry probiotics market size from broilers application should gain above 8%. Growing meat consumption due to disease outbreaks along with government initiatives to promote agribusiness and animal nutrition products will boost regional industry growth. Poultry probiotics market size from poult & chick applications may witness gains at above 6%. Chick & poults need these products to improve gut health and microflora which would improve their immune systems. Prolong exposure towards contaminated environment may cause infections in new born chickens which can be prevented by probiotics which supply complete nutrition and promote digestive metabolism thereby maintaining livestock health. Global poultry probiotics market share is competitive and moderately consolidated. DuPont, Chr. Hansen, Kemin, Novozymes, Lallemand, DSM Evonik, Novus, Biomin, Adddiseo and Prowel are some companies active in this market. Make an inquiry for purchasing this report @ https://www.gminsights.com/inquiry-before-buying/318 Eye Health Ingredients Market size was worth over USD 180 million in 2016 and is set to exceed 1.8 kilo tons by 2024. Hectic & busy life schedules along with exposure to toxic materials have an adverse effect on the eyesight of people leading to suffer from an unhealthy vision. Increasing requirement to maintain the perfect “20/20” eyesight has alleviated the demand for eye health ingredients. https://www.gminsights.com/industry-analysis/probiotics-market Animal Feed Organic Trace Minerals Market size was over USD 490 million in 2016 and will witness CAGR over 6.5% from 2017 to 2024. Upsurge in global meat & dairy production due to its increasing consumption will drive animal feed organic trace minerals market size. The industry will witness a significant gain as trace minerals assist in activating essential hormones and enzymes which are vital for metabolic processes in livestock. Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.


News Article | May 11, 2017
Site: globenewswire.com

Poultry Probiotics Market share is poised to cross USD 1.8 billion by 2024; according to a new research report by Global Market Insights, Inc. Shift in preferences towards using NGPs (natural growth promoters) for livestock farming with regulatory sanctions on AGPs will fuel poultry probiotics market size. Growth in meat & meat products demand due to consumer inclination towards consuming ready-to-eat processed products should drive industry demand. Supporting regulatory norms led by FDA regarding GRAS approval of probiotics along with China promoting product research and product development in feed products for animal nutrition is likely to stimulate industry growth. Request for a sample of this research report @ https://www.gminsights.com/request-sample/detail/318 Germany poultry probiotics market size for layers should witness gains at above 6.5% by 2024. Strict regulatory guidelines by EU Commission regarding antibiotics ban due to bioaccumulation causing resistance to medical antibiotics in animals as well as humans thereby stimulating industry growth in this region. The industry is gaining importance owing to technological and product innovations in this sector which should contribute towards regional growth. Bacillus based poultry probiotics market size should grow significantly to surpass USD 100 million at the end of forecast period. These are widely used in feed products for broiler chicken due to its potential in decreasing cost and concentration of animal feed. Bacillus spores are biocompatible, stable and easily available thereby promoting its use in animal nutrition products. Key feedstock includes Lactobacilli, Bacillus, Streptococcus and Bifidobacterium. These are live bacteria which are hard to maintain and test which is likely to challenge industry profitability. Standardization issue and technological limitations are major limiting factors. Browse key industry insights spread across 175 pages with 157 market data tables & 23 figures & charts from this 2017 report Poultry Probiotics Market in detail along with the table of contents: U.S. poultry probiotics demand for streptococcus may exceed USD 25 million at the end of 2024. Streptococcus plays an important role as growth promoter which helps in increasing the animal weight and enhancing feed efficiency. Prominently used streptococcus strains include S. cristatus, S. thermophiles, and S. faecium. India poultry probiotics market size from broilers application should gain above 8%. Growing meat consumption due to disease outbreaks along with government initiatives to promote agribusiness and animal nutrition products will boost regional industry growth. Poultry probiotics market size from poult & chick applications may witness gains at above 6%. Chick & poults need these products to improve gut health and microflora which would improve their immune systems. Prolong exposure towards contaminated environment may cause infections in new born chickens which can be prevented by probiotics which supply complete nutrition and promote digestive metabolism thereby maintaining livestock health. Global poultry probiotics market share is competitive and moderately consolidated. DuPont, Chr. Hansen, Kemin, Novozymes, Lallemand, DSM Evonik, Novus, Biomin, Adddiseo and Prowel are some companies active in this market. Make an inquiry for purchasing this report @ https://www.gminsights.com/inquiry-before-buying/318 Eye Health Ingredients Market size was worth over USD 180 million in 2016 and is set to exceed 1.8 kilo tons by 2024. Hectic & busy life schedules along with exposure to toxic materials have an adverse effect on the eyesight of people leading to suffer from an unhealthy vision. Increasing requirement to maintain the perfect “20/20” eyesight has alleviated the demand for eye health ingredients. https://www.gminsights.com/industry-analysis/probiotics-market Animal Feed Organic Trace Minerals Market size was over USD 490 million in 2016 and will witness CAGR over 6.5% from 2017 to 2024. Upsurge in global meat & dairy production due to its increasing consumption will drive animal feed organic trace minerals market size. The industry will witness a significant gain as trace minerals assist in activating essential hormones and enzymes which are vital for metabolic processes in livestock. Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.


News Article | May 9, 2017
Site: www.marketwired.com

Florida Predicted to be the Second Largest Cannabis State in the U.S. MIAMI, FL--(Marketwired - May 9, 2017) -  Novus Acquisition and Development, Corp. ( : NDEV) ("Novus"), through its wholly owned subsidiary WCIG Insurance Services, Inc., is a diversified insurance entity in health, liability, annuity and accident, and, the nation's first carrier/aggregator offering a cannabis health plan, today announced that it is permitted to market its health benefits packages in the state of Florida. Novus has begun its entry into what is expected to be the second largest cannabis revenue generating state in the U.S. and become a major player in the cannabis sector that will rival the current markets of Colorado and California. Many analysts project that Florida's cannabis market will grow to $1.6 billion by 2020 at a compound annual growth rate of 140%; making it at least half the size of California's projected $2.6 billion and projecting Florida to be 7.5% of the total legal U.S. cannabis market by 2020. This is supported by many companies coming into Florida, such as publicly-traded Aphria, a Canadian-based cannabis cultivator that plans to purchase and invest $25 million into a Florida cultivation facility that values the state cannabis license at close to $200 million. As Novus continues to invest into the expansion of its infrastructure, this move into Florida will increase its projected 23 million Americans to 30 million by 2018, once markets are fully developed. Novus CEO, Frank Labrozzi, stated, "Novus' business model in the cannabis health insurance is based on owning receivables (insurance policies) rather than depreciative assets such as hard inventory, manufacturing, delivery vehicles and warehousing that all can weigh heavy on a company's intrinsic value. Along with recent announcements that the U.S. Department Of Justice and U.S. Attorney General Jeff Sessions are getting their hands tied by Congress with no funds for the fight against patients that need medical cannabis, this sure points to signs that medical cannabis in the United States is here to stay." Novus and the State of Healthcare from the U.S. Federal Government As it stands many insurance companies, Aetna and Humana for example, are opting out of ObamaCare, leaving millions of Americans without health insurance. This also includes many Americans currently opting out of ObamaCare, for reasons of higher premiums, deductibles, and pharmaceutical coverages. Whether it's ObamaCare or TrumpCare, it does not matter, Novus will continue to design its plans to fill the gaps and seek the inefficiencies and take advantage of the country's current healthcare shortcomings. Novus MedPlan not only covers medical cannabis, but also dental, vision, hearing, diabetic, lab and imaging packages that rival many government-subsidized healthcare plans. In the foreseeable future, Novus plans to announce new benefit packages that will cover many Americans even if ObamaCare is repealed and/or placed. We invite you to review the entire filing here: https://www.otcmarkets.com/stock/NDEV/filings Novus Acquisition & Development Corp. ( : NDEV), through its subsidiary WCIG Insurance, provides health insurance and related insurance solutions within the wellness and medical marijuana industries in states where legal programs exist. Novus has developed its infrastructure within many lines of the insurance business such as, health, property & casualty, life, accident and fixed annuities. Novus medical cannabis benefits package will work as outside developers and will not cultivate, handle, transport grow, extract, dispense, put up for sale, put on the market, vend, deliver, supply, circulate, or trade cannabis or any substances that violate the United States law or the Controlled Substances Act, nor does it intend to do so in the future and will continue to follow state and federal laws. The statements made about specific products have not been evaluated by the United States Food and Drug Administration (FDA) and are not intended to diagnose, treat, cure or prevent disease. All information provided on these press releases or any information contained on or in any product label or packaging is for informational purposes only and is not intended as a substitute for advice from your physician or other health care professional. Once a push notification is competed the transaction is solely between the state-licensed dispensary and the registered patient. The state laws are in conflict with the federal Controlled Substances Act. The current administration has effectively stated that it is not an efficient use of resources to direct federal law enforcement agencies to prosecute those lawfully abiding by state designated laws, allowing the use and distribution of medical marijuana. However, there is no guarantee that the current administration, nor any future administration, will not change this policy and decide to enforce the federal laws strongly. Any such change in the federal government's enforcement of current federal laws could cause significant financial changes to Novus Medical Group. While we do not intend to harvest, distribute or sell cannabis or cannabis related products, we may be harmed by a change in enforcement by federal or state governments. For more information, please visit: http://www.getnovusnow.com This release includes forward-looking statements, which are based on certain assumptions and reflects management's current expectations. These forward-looking statements are subject to a number of risks and uncertainties that could cause actual results or events to differ materially from current expectations. Some of these factors include: general global economic conditions; general industry and market conditions and growth rates; uncertainty as to whether our strategies and business plans will yield the expected benefits; increasing competition; availability and cost of capital; the ability to identify and develop and achieve commercial success; the level of expenditures necessary to maintain and improve the quality of services; changes in the economy; changes in laws and regulations, includes codes and standards, intellectual property rights, and tax matters; or other matters not anticipated; our ability to secure and maintain strategic relationships and distribution agreements. Novus Medical Group disclaims any intention or obligation to update or revise any forward-looking statements, whether as a result of new information, future events or otherwise.


News Article | May 11, 2017
Site: globenewswire.com

Poultry Probiotics Market share is poised to cross USD 1.8 billion by 2024; according to a new research report by Global Market Insights, Inc. Shift in preferences towards using NGPs (natural growth promoters) for livestock farming with regulatory sanctions on AGPs will fuel poultry probiotics market size. Growth in meat & meat products demand due to consumer inclination towards consuming ready-to-eat processed products should drive industry demand. Supporting regulatory norms led by FDA regarding GRAS approval of probiotics along with China promoting product research and product development in feed products for animal nutrition is likely to stimulate industry growth. Request for a sample of this research report @ https://www.gminsights.com/request-sample/detail/318 Germany poultry probiotics market size for layers should witness gains at above 6.5% by 2024. Strict regulatory guidelines by EU Commission regarding antibiotics ban due to bioaccumulation causing resistance to medical antibiotics in animals as well as humans thereby stimulating industry growth in this region. The industry is gaining importance owing to technological and product innovations in this sector which should contribute towards regional growth. Bacillus based poultry probiotics market size should grow significantly to surpass USD 100 million at the end of forecast period. These are widely used in feed products for broiler chicken due to its potential in decreasing cost and concentration of animal feed. Bacillus spores are biocompatible, stable and easily available thereby promoting its use in animal nutrition products. Key feedstock includes Lactobacilli, Bacillus, Streptococcus and Bifidobacterium. These are live bacteria which are hard to maintain and test which is likely to challenge industry profitability. Standardization issue and technological limitations are major limiting factors. Browse key industry insights spread across 175 pages with 157 market data tables & 23 figures & charts from this 2017 report Poultry Probiotics Market in detail along with the table of contents: U.S. poultry probiotics demand for streptococcus may exceed USD 25 million at the end of 2024. Streptococcus plays an important role as growth promoter which helps in increasing the animal weight and enhancing feed efficiency. Prominently used streptococcus strains include S. cristatus, S. thermophiles, and S. faecium. India poultry probiotics market size from broilers application should gain above 8%. Growing meat consumption due to disease outbreaks along with government initiatives to promote agribusiness and animal nutrition products will boost regional industry growth. Poultry probiotics market size from poult & chick applications may witness gains at above 6%. Chick & poults need these products to improve gut health and microflora which would improve their immune systems. Prolong exposure towards contaminated environment may cause infections in new born chickens which can be prevented by probiotics which supply complete nutrition and promote digestive metabolism thereby maintaining livestock health. Global poultry probiotics market share is competitive and moderately consolidated. DuPont, Chr. Hansen, Kemin, Novozymes, Lallemand, DSM Evonik, Novus, Biomin, Adddiseo and Prowel are some companies active in this market. Make an inquiry for purchasing this report @ https://www.gminsights.com/inquiry-before-buying/318 Eye Health Ingredients Market size was worth over USD 180 million in 2016 and is set to exceed 1.8 kilo tons by 2024. Hectic & busy life schedules along with exposure to toxic materials have an adverse effect on the eyesight of people leading to suffer from an unhealthy vision. Increasing requirement to maintain the perfect “20/20” eyesight has alleviated the demand for eye health ingredients. https://www.gminsights.com/industry-analysis/probiotics-market Animal Feed Organic Trace Minerals Market size was over USD 490 million in 2016 and will witness CAGR over 6.5% from 2017 to 2024. Upsurge in global meat & dairy production due to its increasing consumption will drive animal feed organic trace minerals market size. The industry will witness a significant gain as trace minerals assist in activating essential hormones and enzymes which are vital for metabolic processes in livestock. Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.


News Article | May 11, 2017
Site: globenewswire.com

Poultry Probiotics Market share is poised to cross USD 1.8 billion by 2024; according to a new research report by Global Market Insights, Inc. Shift in preferences towards using NGPs (natural growth promoters) for livestock farming with regulatory sanctions on AGPs will fuel poultry probiotics market size. Growth in meat & meat products demand due to consumer inclination towards consuming ready-to-eat processed products should drive industry demand. Supporting regulatory norms led by FDA regarding GRAS approval of probiotics along with China promoting product research and product development in feed products for animal nutrition is likely to stimulate industry growth. Request for a sample of this research report @ https://www.gminsights.com/request-sample/detail/318 Germany poultry probiotics market size for layers should witness gains at above 6.5% by 2024. Strict regulatory guidelines by EU Commission regarding antibiotics ban due to bioaccumulation causing resistance to medical antibiotics in animals as well as humans thereby stimulating industry growth in this region. The industry is gaining importance owing to technological and product innovations in this sector which should contribute towards regional growth. Bacillus based poultry probiotics market size should grow significantly to surpass USD 100 million at the end of forecast period. These are widely used in feed products for broiler chicken due to its potential in decreasing cost and concentration of animal feed. Bacillus spores are biocompatible, stable and easily available thereby promoting its use in animal nutrition products. Key feedstock includes Lactobacilli, Bacillus, Streptococcus and Bifidobacterium. These are live bacteria which are hard to maintain and test which is likely to challenge industry profitability. Standardization issue and technological limitations are major limiting factors. Browse key industry insights spread across 175 pages with 157 market data tables & 23 figures & charts from this 2017 report Poultry Probiotics Market in detail along with the table of contents: U.S. poultry probiotics demand for streptococcus may exceed USD 25 million at the end of 2024. Streptococcus plays an important role as growth promoter which helps in increasing the animal weight and enhancing feed efficiency. Prominently used streptococcus strains include S. cristatus, S. thermophiles, and S. faecium. India poultry probiotics market size from broilers application should gain above 8%. Growing meat consumption due to disease outbreaks along with government initiatives to promote agribusiness and animal nutrition products will boost regional industry growth. Poultry probiotics market size from poult & chick applications may witness gains at above 6%. Chick & poults need these products to improve gut health and microflora which would improve their immune systems. Prolong exposure towards contaminated environment may cause infections in new born chickens which can be prevented by probiotics which supply complete nutrition and promote digestive metabolism thereby maintaining livestock health. Global poultry probiotics market share is competitive and moderately consolidated. DuPont, Chr. Hansen, Kemin, Novozymes, Lallemand, DSM Evonik, Novus, Biomin, Adddiseo and Prowel are some companies active in this market. Make an inquiry for purchasing this report @ https://www.gminsights.com/inquiry-before-buying/318 Eye Health Ingredients Market size was worth over USD 180 million in 2016 and is set to exceed 1.8 kilo tons by 2024. Hectic & busy life schedules along with exposure to toxic materials have an adverse effect on the eyesight of people leading to suffer from an unhealthy vision. Increasing requirement to maintain the perfect “20/20” eyesight has alleviated the demand for eye health ingredients. https://www.gminsights.com/industry-analysis/probiotics-market Animal Feed Organic Trace Minerals Market size was over USD 490 million in 2016 and will witness CAGR over 6.5% from 2017 to 2024. Upsurge in global meat & dairy production due to its increasing consumption will drive animal feed organic trace minerals market size. The industry will witness a significant gain as trace minerals assist in activating essential hormones and enzymes which are vital for metabolic processes in livestock. Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.


News Article | May 11, 2017
Site: globenewswire.com

Poultry Probiotics Market share is poised to cross USD 1.8 billion by 2024; according to a new research report by Global Market Insights, Inc. Shift in preferences towards using NGPs (natural growth promoters) for livestock farming with regulatory sanctions on AGPs will fuel poultry probiotics market size. Growth in meat & meat products demand due to consumer inclination towards consuming ready-to-eat processed products should drive industry demand. Supporting regulatory norms led by FDA regarding GRAS approval of probiotics along with China promoting product research and product development in feed products for animal nutrition is likely to stimulate industry growth. Request for a sample of this research report @ https://www.gminsights.com/request-sample/detail/318 Germany poultry probiotics market size for layers should witness gains at above 6.5% by 2024. Strict regulatory guidelines by EU Commission regarding antibiotics ban due to bioaccumulation causing resistance to medical antibiotics in animals as well as humans thereby stimulating industry growth in this region. The industry is gaining importance owing to technological and product innovations in this sector which should contribute towards regional growth. Bacillus based poultry probiotics market size should grow significantly to surpass USD 100 million at the end of forecast period. These are widely used in feed products for broiler chicken due to its potential in decreasing cost and concentration of animal feed. Bacillus spores are biocompatible, stable and easily available thereby promoting its use in animal nutrition products. Key feedstock includes Lactobacilli, Bacillus, Streptococcus and Bifidobacterium. These are live bacteria which are hard to maintain and test which is likely to challenge industry profitability. Standardization issue and technological limitations are major limiting factors. Browse key industry insights spread across 175 pages with 157 market data tables & 23 figures & charts from this 2017 report Poultry Probiotics Market in detail along with the table of contents: U.S. poultry probiotics demand for streptococcus may exceed USD 25 million at the end of 2024. Streptococcus plays an important role as growth promoter which helps in increasing the animal weight and enhancing feed efficiency. Prominently used streptococcus strains include S. cristatus, S. thermophiles, and S. faecium. India poultry probiotics market size from broilers application should gain above 8%. Growing meat consumption due to disease outbreaks along with government initiatives to promote agribusiness and animal nutrition products will boost regional industry growth. Poultry probiotics market size from poult & chick applications may witness gains at above 6%. Chick & poults need these products to improve gut health and microflora which would improve their immune systems. Prolong exposure towards contaminated environment may cause infections in new born chickens which can be prevented by probiotics which supply complete nutrition and promote digestive metabolism thereby maintaining livestock health. Global poultry probiotics market share is competitive and moderately consolidated. DuPont, Chr. Hansen, Kemin, Novozymes, Lallemand, DSM Evonik, Novus, Biomin, Adddiseo and Prowel are some companies active in this market. Make an inquiry for purchasing this report @ https://www.gminsights.com/inquiry-before-buying/318 Eye Health Ingredients Market size was worth over USD 180 million in 2016 and is set to exceed 1.8 kilo tons by 2024. Hectic & busy life schedules along with exposure to toxic materials have an adverse effect on the eyesight of people leading to suffer from an unhealthy vision. Increasing requirement to maintain the perfect “20/20” eyesight has alleviated the demand for eye health ingredients. https://www.gminsights.com/industry-analysis/probiotics-market Animal Feed Organic Trace Minerals Market size was over USD 490 million in 2016 and will witness CAGR over 6.5% from 2017 to 2024. Upsurge in global meat & dairy production due to its increasing consumption will drive animal feed organic trace minerals market size. The industry will witness a significant gain as trace minerals assist in activating essential hormones and enzymes which are vital for metabolic processes in livestock. Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.


No statistical methods were used to predetermine sample size. The experiments were not randomized, and investigators were not blinded to allocation during experiments and outcome assessment. Recombinant adenoviruses were constructed with the following inserts. Full-length mouse Dkk1 (ref. 6), and mouse Rspo1-Fc16 with full-length Rspo1 fused to a mouse antibody IgG2α Fc fragment at the C terminus have been described. Human RSPO2 and mouse Rnf43 ECD and Znrf3 ECD similarly contained full-length open reading frames with a C-terminal mouse IgG2α Fc fragment. Mouse Fzd8 CRD (residues 25–173) was cloned with an N-terminal haemagglutinin (HA) epitope tag and C-terminal IgG2α Fc fragment. In addition, a recombinant adenovirus was engineered to express human LGR5 ECD with both C-terminal FLAG and histidine tags. The construction of the adenoviruses encoding the scFv–DKK1c Wnt surrogate agonist and scFv–DKK1c–RSPO2 single-chain polypeptide fusion, each with a C-terminal His tag is described in a companion paper by Janda et al.25 On day 2 after intravenous injection, scFv–DKK1c was found to be expressed in vivo at ~10–20 μg ml−1 (280–560 nM) in mouse sera and the serum potently induced TOPflash activity in vitro. Full-length Wnt3a cDNA (a gift from R. Nusse) was cloned without any epitope tags and detected by western blotting with anti-WNT3A (Cell Signaling 2391) against a recombinant WNT3A protein. No detectable WNT3A protein was found in mouse sera after intravenous injection. All adenoviral constructs contained an N-terminal signal peptide sequence to allow for their secretion. These adenoviruses were cloned by homologous recombination into E1− E3− adenovirus strain 5, purified by double CsCl gradient, and titred as previously described33. Recombinant proteins were expressed in serum-free CD293 medium (Invitrogen) of HEK293 cells infected by adenovirus. Recombinant LGR5-ECD protein was purified by nickel-NTA affinity chromatography (Qiagen) from Ad-LGR5-ECD-infected CD293 medium. Likewise, recombinant RNF43 and ZNRF3 ECD-Fc fusion proteins were purified by protein A affinity chromatography (KPL) from Ad-Rnf43-ECD-infected or Ad-Znrf3-ECD-infected CD293 medium, respectively. Protein purity was verified by Coomassie-stained SDS–PAGE. Adult Lgr5-eGFP-IRES-creER mice7 (Jax) or Axin2-LacZ mice (Jax) between 8 and 12 weeks old were injected intravenously with adenoviruses (doses of 5 × 108 to 1 × 109 pfu per mouse). Lgr5-eGFP-IRES-creER mice were crossed with Rosa26-tdTomato mice to generate Lgr5-eGFP-IRES-creER; Rosa26-tdTomato compound heterozygous mice. Similarly, Villin-creER or Actin-creER mice were crossed to Rosa26-Rainbow mice to generate Villin-creER; Rosa26-Rainbow or Actin-creER; Rosa26-Rainbow compound heterozygous mice. Mice were dosed with adenoviruses as above, and serum expression of all ECDs was confirmed by immunoblotting and histological assessment of intestinal crypt hyperplasia for those treated with Ad-Rspo1 and Ad-RSPO2. Adult mice between 8 and 12 weeks of age were administered tamoxifen (Sigma) dosed at 4 mg per 40 g body weight to genetically label for lineage tracing experiments using the various Rosa26 reporter strains. All in vivo experiments used n = 3–5 mice per group and were repeated at least twice except for the RNA-seq studies. Both male and female mice were used. All animal experiments were conducted in accordance with procedures approved by the IACUC at Stanford University. FACS experiments were performed using fresh small intestine epithelial preparations. A standardized 3 cm segment of proximal jejunum was used for quantitative FACS analysis of ISC populations. Intestinal epithelial cells were extracted from en bloc resected small intestine with 10 mM EDTA and manual shaking, followed by enzymatic dissociation with collagenase/dispase (Roche) to generate a single-cell suspension. Singlet discrimination was sequentially performed using plots for forward scatter (FSC-A versus FSC-H) and side scatter (SSC-W versus SSC-H). Dead cells were excluded by scatter characteristics and viability stains. All FACS experiments were performed on an Aria II sorter (BD) or LSRII analyser (BD) at the Stanford University Shared FACS Facility and FACS data were analysed using FlowJo software (TreeStar). Intestinal tissue was collected and fixed in 4% paraformaldehyde. 8-μm OCT frozen sections or 5-μm paraffin-embedded sections were TUNEL-stained using the DeadEnd Fluorometric TUNEL system per manufacturer’s instructions (Promega) or immunostained using the following primary antibodies: anti-Ki67 (ThermoFisher RM-9106), anti-MUC2 (Santa Cruz sc-15334), anti-lysozyme (Dako A0099), anti-chromogranin A (Santa Cruz sc-1488), anti-FABP1 (Novus NBP1-87695), anti-CD44 (BD Pharmingen 550538), anti-cyclin D1 (Abcam ab134175) and anti-CD166 (R&D AF1172). All primary antibodies were used at 1:100 to 1:200 dilutions. Cy3- and Cy5-conjugated secondary antibodies (Santa Cruz and Jackson ImmunoResearch) were used at 1:500 to 1:1,000 dilutions. Alexa Fluor 594-conjugated phalloidin (Invitrogen) was used at 1:500. CD166 immunostained tissue sections34 were analysed and confocal images acquired as 0.5-μm planes using an IX81 Inverted Microscope equipped with Fluoview FV1000-Spinning Disc Confocal scan head and FV10 ASW 1.7 software (Olympus). All other images were captured on a Zeiss Axio-Imager Z1 with ApoTome or Leica SP5 confocal microscope. In situ hybridization for Olfm4 mRNA was performed using the RNAscope kit (Advanced Cell Diagnostics) according to the manufacturer’s instructions. In brief, 5 μm formalin-fixed, paraffin-embedded tissue sections or 8 μm OCT frozen sections were pre-treated with heat and protease before hybridization with a target probe to Olfm4 mRNA. A horseradish peroxidase (HRP)-based signal amplification system was then hybridized to the target probes followed by colorimetric development with DAB. Negative control probes for the bacterial gene DapB were also included for each slide. Adult Lgr5-eGFP-IRES-creER mice (Jax) between 10 and 12 weeks old were treated with intravenous adenovirus. After 48 h, these mice were treated by oral gavage for 4 days with twice daily dosing interval with either 50 mg kg−1 of PORCN inhibitor C59 (Cellagen Technology) or vehicle consisting of 0.5% methylcellulose plus 0.1% Tween80, as previously described35. Mice were euthanized 20 h after the last dose of C59 and the intestine was harvested for FACS and histological analysis. Small intestine tissue samples were fixed with 2.5% glutaraldehyde and post-fixed in 1% osmium tetroxide in 100 mM phosphate buffer. Tissue was dehydrated, embedded in epoxy resin, and visualized by a JEOL transmission electron microscope at 120 kV (model JEM-1210). L cells stably transfected with TOPflash dual reporter plasmid system (a gift from J. Chen) were used in TOPflash dual luciferase assays (Promega Dual Luciferase kit) with WNT3A conditioned medium from a stably transfected WNT3A-expressing cell line (a gift from R. Nusse) from which activation of the TOPflash reporter has been confirmed; mycoplasma contamination was not tested. Recombinant WNT3A (R&D) was alternatively used. Recombinant mouse RSPO1–RSPO4 proteins (R&D) were used at 5 pM concentration each in these assays. Recombinant LGR5, RNF43 and ZNRF3 ECD proteins were expressed and purified as above and their purity and protein concentrations were determined by Coomassie-stained SDS–PAGE and Bradford assays. Assays were visualized with a Tecan M1000 luminometer. Recombinant scFv–DKK1c was expressed and purified as described in the companion paper25. The kinetics and affinity of interactions between RSPO1–RSPO4 and Flag- and histidine-tagged LGR5 ECD, Fc-tagged RNF43 ECD or Fc-tagged ZNRF3 ECD were determined by surface plasmon resonance. Data were collected on the BIAcore T100 instrument (GE Healthcare). Approximately 1,000 resonance units (RU) of recombinant mouse RSPO1, RSPO2, RSPO3 or RSPO4 (R&D) were immobilized on a CM5 sensor chip (GE Healthcare) using standard amine coupling. Increasing concentrations of LGR5 ECD, RNF43 ECD or ZNRF3 ECD were passed over the chip in HBS supplemented with 0.005% surfactant P20 (HBS+P). Binding phases for the LGR5-ECD were performed at 50 μl min−1 for 240 s and dissociation phases were performed at 50 μl min−1 for 1,850 s. The chip was regenerated after each injection with 240-s washes with 0.5 M magnesium chloride. Binding and dissociation phases for RNF43 ECD and ZNRF3 ECD were each performed at 50 μl min−1 for 120 s. The chip was regenerated after each injection with 120-s washes with 1 M magnesium chloride. All curves were reference-subtracted from a flow cell containing 1,000 RU of a negative control protein (hen egg white lysozyme or BSA). Curves were fitted using the BIAcore T100 evaluation software to a 1:1 model to determine the association rate (k ), dissociation rate (k ) and dissociation constant (K ). The kinetics and affinity of anti-RSPO antibody interactions with RSPO1–RSPO4 were determined as described for RNF43 and ZNRF3, except that the regeneration buffer was 25% ethylene glycol and 2.25 M magnesium chloride. The kinetics and affinity of Fc-tagged RNF43 and ZNRF3 ECDs are enhanced by avidity effects due to Fc-dimerization. The furin 1 and 2 repeats of human RSPO2 were cloned into the pCT302 vector as a C-terminal fusion to a c-Myc epitope and the cell-wall protein AGA2. RSPO2 was displayed on the EBY100 strain of Saccharomyces cerevisiae as previously described36. Competent yeast cells were electroporated with the RSPO2 expression plasmid and recovered in SDCAA selection media. The cultures were harvested in log phase, and yeast cells were then pelleted and resuspended in SGCAA induction media. Surface expression of RSPO2 was detected by staining yeast with a 488-labelled antibody to the c-Myc epitope (Cell Signaling 279), and then analysed by flow cytometry. Binding of LGR5, RNF43 and ZNRF3 ECDs was tested by incubating yeast with 200 nM recombinant Flag-tagged LGR5 ECD or with Fc-tagged RNF43 ECD or ZNRF3 ECD in PBS and 0.1% BSA for 2 h, washing twice with PBS and 0.1% BSA and then incubating for 30 min with an Alexa Fluor 647-labelled antibody to the Flag epitope (Cell Signaling 3916S) (for LGR5 binding) or a PE-labelled anti-IgG antibody (eBioscience 12-4998-82). Cells were washed twice with PBS and 0.1% BSA and then analysed by flow cytometry. Sequential staining of yeast was performed by incubating samples with 200 nM LGR5-ECD, 200 nM RNF43 ECD, or 200 nM ZNRF3 ECD alone, washing and then incubating with a mixture of (200 nM LGR5-ECD and 200 nM RNF43-ECD) or (200 nM LGR5-ECD and 200 nM ZNRF3 ECD). Cells double-stained with both LGR5-ECD and either RNF43 or ZNRF3 ECD were then washed and incubated with a mixture of PE-anti-IgG and 647-anti-Flag before a final wash and analysis by flow cytometry. Cells were isolated by flow cytometry into RNEasy lysis buffer (Qiagen) from n = 2–3 mice per condition, 1.5 days after injection of the appropriate adenoviruses. A 1.8× volume of AMPure beads (Beckman Coulter) was added to the thawed cell lysates. After a 30-min incubation at room temperature, the samples were washed twice with 70% ethanol and eluted in 22 μl water. The samples were then digested with 0.6 mAU Proteinase K (Qiagen) in the presence of 1× NEB buffer 1 (NEB) at 50 °C for 20 min, followed by a heat-inactivation step at 65 °C for 10 min. A DNase digestion was performed using the RNase-Free DNase Set (Qiagen) at 37 °C for 30 min. The samples were cleaned with a 1.8× volume of AMPure XP beads (Beckman Coulter). 1 ng of purified total RNA, as determined by Agilent Bioanalyzer (Agilent Technologies), was processed with the mRNA direct micro kit (Life Technologies) to select for poly A RNA. Each entire sample was input into the Ambion WT Expression Kit (Life Technologies) to perform double-stranded cDNA synthesis followed by in vitro transcription to generate amplified cRNA. The cRNA was purified following the manufacturer’s instructions and the concentration was determined with a NanoDrop instrument (ThermoFisher). 1 μg of cRNA was fragmented in 1× fragmentation buffer (mRNA-Seq Sample Prep Kit, Illumina) at 94 °C for 5 min, then placed on ice and the reaction was stopped by the addition of 20 mM EDTA. The fragments were precipitated with 70 mM sodium acetate (Life Technologies), 40 μg glycogen (Life Technologies) and 70% ethanol at −80 °C for 1 h followed by centrifugation and washing with 70% ethanol. 3 μg of random hexamer (Life Technologies) was added to the fragmented, purified cRNA and incubated at 70 °C for 10 min to anneal the primer. The first strand reaction was performed with 200 units of SuperScript II (Life Technologies) with 0.625 mM dNTPs (NEB) and 8U SUPERase RNase Inhibitor (Life Technologies) at 25 °C for 10 min, then 42 °C for 50 min, then 75 °C for 15 min and cooled to 4 °C. In second-strand synthesis, 1× second strand buffer (Illumina) and 0.3 mM dNTPs (Illumina) were added and the samples were incubated at 4 °C for 5 min before adding 50 U of DNA Polymerase (NEB) and 5 U of Rnase H (NEB). The samples were mixed well and incubated at 16 °C for 2.5 h, followed by purification with the MinElute Kit (Qiagen). To perform library prep, the samples were end repaired using a Quick Blunting Kit (NEB) and incubated at 20 °C for 1 h, then 75 °C for 30 min to inactivate the enzyme. To produce overhangs aimed to improve subsequent ligation efficiency, a single A base was added to the 3′ ends of each fragment with 2 mM dATP and 5 units of Klenow fragment 3′-5′ exo- DNA Polymerase (NEB) at 37 °C for 45 min, followed by 75 °C for 30 min to inactivate the enzyme. Using a quick ligase kit (NEB), 0.5 μM of adaptors containing single T base overhangs were ligated to the cDNA fragments at 12 °C for 75 min, then 80 °C for 20 min and cooled to 4 °C. These adaptors contain barcodes to facilitate sample multiplexing during sequencing. The adaptor sequence is preceded by four random nucleotides to add diversity to the pooled library. The samples were pooled by combining 5 μl of each library. After AMPure XP cleanup, one-half of the pooled library was run on the Pippin Size Selection Instrument (Sage Sciences) to select for 200 bp fragments. Library amplification was performed on one-half of the Pippin eluate in 1× Phusion GC buffer with 0.2 mM dNTPs, 0.1 μM forward primer (IDT), 0.1 μM reverse primer, 1 U Phusion Hot Start II Polymerase (Thermo Fisher Scientific). The reaction was run with the following program: 98 °C for 30 s, then 15 cycles of 98 °C for 10 s, 65 °C for 30 s, 72 °C for 30 s, then 72 °C for 4 min and cooled to 4 °C. The amplified library was cleaned using a 1× volume of AMPure XP beads and QC was run with the Agilent Bioanalyzer DNA 1000 kit, followed by concentration determination by qPCR using the KAPA Library Quantification Kit (KAPA Biosystems). To perform sequencing, the library was diluted to 4 nM and denatured with 0.1 N NaOH. Following denaturation, the library was further diluted to 4 pM and run on the Illumina HiSeq 2500 in paired-end, 100 × 100 bp format. Sequenced reads were aligned to the mouse reference genome mm9 (UCSC) using TopHat37 with the transcript annotation supplied. The mapped reads was assigned to gene using the tool htseq-count of the Python package HTseq38, with the default union-counting mode. The output of htseq-count was used as input for DESeq2 (ref. 39) to perform differential expression analysis, with a false discovery rate (FDR) of 10% as the cutoff. In addition, a filtering criterion of mean fragments per kilobase of transcript per million mapped reads (FPKM) of 1 in at least one condition was used to define expressed transcripts in each differential expression analysis. Cufflinks40 was used to calculate gene count and perform FPKM normalization. Gene Ontology term analysis was performed using DAVID functional annotation tool41. A FDR of 10% was applied to evaluate the significance. Lgr5-eGFP-IRES-creER mice were treated with adenovirus in vivo, and then 26 h after treatment the proximal jejunum was harvested to generate a single-cell suspension and FACS isolated using the endogenous GFP signal, as above. The sorted cellular suspensions were loaded on a GemCode Single Cell Instrument (10x Genomics) to generate single-cell gel beads in emulsion (GEMs). Approximately 1,200–2,800 cells were loaded per channel. Two technical replicates were generated per sorted cell suspension. Single-cell RNA-seq libraries were prepared using GemCode Single Cell 3′ Gel Bead and Library Kit (now sold as P/N 120230, 120231, 120232, 10x Genomics) as described previously29. Sequencing libraries were loaded at 2.1 pM on an Illumina Next-Seq500 with 2 × 75 paired-end kits using the following read length: 98 bp read1, 14 bp I7 index, 8 bp I5 index and 5 bp read2. Note that these libraries were generated before the official launch of GemCode Single Cell 3′ Gel Bead and Library Kit. Thus, 5 bp UMI was used (the official GemCode Single Cell 3′ Gel Bead contains 10 bp UMI). The Cell Ranger Single Cell Software Suite was used to perform sample de-multiplexing, barcode processing, and single-cell 3′ gene counting (http://software.10xgenomics.com/single-cell/overview/welcome). 5 bp UMI tags were extracted from read2. We analysed a total of 13,247 single cells, consisting of 11,268 FACS-sorted Lgr5–eGFP+and 1,979 Ad-Fc-treated Lgr5–eGFP− cells. Two technical replicates (the number of cells recovered per channel ranges from around 400 to 1,400 cells) were generated from each treatment condition. The mean raw reads per cell varied from ~45 k to 86 k. Each sample was downsampled to 28,439 confidently mapped reads per cell. Then the gene-cell barcode matrix from each sample was concatenated. The gene-cell barcode matrix was filtered based on number of genes detected per cell (any cells with less than 400 or more than 4,400 genes per cell were filtered) and percentage of mitochondrial UMI counts (any cells with more than 10% of mitochondrial UMI counts were filtered). Altogether, 13,176 cells, and 15,865 genes were kept for analysis by the Seurat R package30. Among these 13,176 cells, 74 did not show any epithelial cell markers so they were removed leaving a final total of 13,102 cells, consisting of 1,925 Ad-Fc-treated Lgr5–eGFP− cells and 11,177 Lgr5–eGFP+ cells across six conditions. 2,289 variable genes were selected based on their expression and dispersion (expression cutoff = 0.0125, and dispersion cutoff = 0.5). The first 11 principal components were used for the t-SNE projection and clustering analysis (resolution = 0.3, k.seed = 100). We applied sSeq from ref. 42 to identify genes that are enriched in a specific cluster (the specific cluster is assigned as group a, and the rest of clusters is assigned as group b). There are a few differences between our implementation and ref. 42. First, we used the ratio of total UMI counts and median of total UMI counts across all cells as the size factors. Second, the quantile rule of thumb was used to estimate the shrinkage target. Third, for genes with large counts, an asymptotic approximation from the edgeR package43 was used instead of the negative binomial exact test to speed up the computation. For the heatmap in Extended Data Fig. 9h, the gene list was furthered filtered requiring minimum UMI counts of 5 in each group, with a positive log fold change of mean expression between the two groups, and an adjusted P < 0.01. The top 10 genes specific to each cluster were picked, and their mean expression was centre scaled before used for the heatmap. Classification of cells was inferred from the annotation of cluster-specific genes. The stem cell clusters (clusters 0 and 1) were marked by enrichment of Lgr5, Olfm4 and Ascl2. Non-cycling and cycling stem cells were distinguished by the enrichment of cell cycle markers such as Mki67 and Tuba1b. Transit amplifying cells (cluster 2) were classified based on the enrichment of cell cycle markers and lack of Lgr5+ stem-cell marker expression. Enterocytes (clusters 3 and 4) were annotated based on the enrichment of markers such as Alpi and Reg1 and prior studies31. Goblet cells (cluster 5) were annotated based on the enrichment of markers such as Muc2 and Guca2a. Paneth cells (cluster 6) were annotated based on the enrichment of Defa genes. Tuft cells (cluster 7) were annotated based on the enrichment of markers such as Dclk1. EE cells (cluster 8) were annotated based on the enrichment of markers such as Chga and Chgb. To compare the global expression difference between samples and the Fc control, we first normalized gene expression by the sum of their UMI counts across all cells in the sample (adding 1 to the numerator and denominator to avoid dividing by 0 for genes that were not detected at all). Then we compared the normalized gene expression between the samples and the Fc control. To generate the heatmap, we furthered filtered the gene list: (1) Only genes with UMI counts >2 in each sample and a log fold change of >1 were considered. (2) The top 15 up- or downregulated genes were picked per sample–Fc comparison, and the union of all genes was used for the heatmap. Data generated during this study are available in the Gene Expression Omnibus (GEO) repository under accession numbers GSE92377 and GSE92865. All other data are available from the corresponding author upon reasonable request.


Patent
Novus Inc | Date: 2013-11-05

A system, method, and computer program product for modeling a risk are provided. An equalizer and an equalizer backend model portfolio risk based on a scenario generated by the equalizer. A scenario includes an active factor, a passive factor, and a change in an active factor level. To model portfolio risk, the equalizer backend uses two-tiered regression analysis. In the first regression, the equalizer backend regresses the passive factor against the active factor and determines changes in a passive factor level. In the second regression, the equalizer backend regresses the changes in the passive and active factor levels against positions in the portfolio that models portfolio risk. The equalizer displays the modeled portfolio risk.

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