#Adult Bovine Serum

[poonamcmi]

Global Adult Bovine Serum Market Is Estimated To Witness High Growth Owing To Increasing Demand for Cell Culture Applications

The global adult bovine serum market is estimated to be valued at US$ 119.3 million in 2022 and is expected to exhibit a CAGR of 5.6% over the forecast period 2022-2030, as highlighted in a new report published by Coherent Market Insights. Market Overview: Adult bovine serum is a high-quality supplement used in various cell culture applications. It provides essential nutrients, growth factors, and hormones required for cell growth and proliferation. The increasing demand for adult bovine serum can be attributed to its advantages such as better cell attachment, faster cell growth rate, and higher cell yield. These serum products are extensively used in academic research, pharmaceutical and biotechnology industries, and in vitro diagnostics. Market Key Trends: One key trend in the global adult bovine serum market is the growing adoption of serum-free culture media. Serum-free culture media eliminates potential contamination issues associated with traditional serum-based cultures. It also allows for increased control over the specific components used in the media, resulting in more consistent and reproducible results. The use of serum-free culture media is particularly beneficial in applications that require high precision and accuracy, such as drug discovery and development. For example, Thermo Fisher Scientific offers Gibco™ AIM V® Medium, a proprietary serum-free culture medium, designed for the expansion of multiple cell types including T-cells and dendritic cells. This medium enables robust cell growth and function without the use of animal-derived serum. PEST Analysis: - Political: The use of animal-derived products, including bovine serum, is subject to regulations and guidelines set by government bodies. Regulatory compliance plays a crucial role in the market dynamics of adult bovine serum. - Economic: The adult bovine serum market is driven by the increasing investment in research and development activities by pharmaceutical and biotechnology companies. - Social: The growing focus on personalized medicine and regenerative therapies is fueling the demand for advanced cell culture applications, driving the growth of the adult bovine serum market. - Technological: Advancements in cell culture technologies and the development of serum-free culture media are shaping the market landscape of adult bovine serum. Key Takeaways: 1: The global Adult Bovine Serum Market Growth is expected to witness high growth, exhibiting a CAGR of 5.6% over the forecast period, due to increasing demand for cell culture applications. The advantages of adult bovine serum, such as better cell attachment, faster cell growth rate, and higher cell yield, are driving its adoption in various industries. 2: Regionally, North America is the fastest-growing and dominating region in the global adult bovine serum market. The presence of key players and the high investment in research and development activities are contributing to the market growth in this region. 3: Key players operating in the global adult bovine serum market include HiMedia Laboratories, Sartorius AG, Thermo Fisher Scientific, Auckland BioSciences Ltd., Moregate Biotech, Bovogen Biologicals, Merck KGaA, Capricorn Scientific GmbH, Gemini Bio, ROCKY MOUNTAIN BIOLOGICALS, Otto Chemie Pvt. Ltd, CellSera Australia, Serana Europe GmbH, TRINA BIOREACTIVES AG, and WISENT BIOPRODUCTS. These companies are focusing on product development and strategic collaborations to strengthen their market presence. In conclusion, the global adult bovine serum market is projected to experience significant growth in the coming years due to the increasing demand for cell culture applications. The adoption of serum-free culture media and advancements in cell culture technologies are expected to contribute to the market expansion. However, regulatory compliance and the emergence of alternative cell culture techniques may pose challenges to market growth.

[reality-detective]

“I gathered all vaccine ingredients into a list and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of that conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... But I'd have to easily say that they're all toxic to humans... Used in fertilizers... Pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... But you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... These are the ingredients used in vaccines... With binding agents to make sure the body won't flush these out... To keep the antibody levels up indefinitely...

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~  By  Iris Figueroa

Here are just SOME vaccine ingredients present in routine vaccines:

◾️Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

◾️Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

◾️Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

◾️Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

◾️Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

◾️Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

◾️Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

◾️Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

◾️Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

◾️African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

◾️Acetone - Can cause kidney, liver, and nerve damage.

◾️E.Coli - Yes, you read that right.

◾️DNA from porcine (pig) Circovirus type-1

◾️Human embryonic lung cell cultures (from aborted fetuses)

You can view all of these ingredients on the CDCs website: 👇

You are always welcome to do your own research, in fact I encourage you to do so. 🤔

[moleculardepot]

Lipids Cholesterol Rich from Adult Bovine Serum

Lipids Cholesterol Rich from Adult Bovine Serum Catalog number: B2015452 Lot number: Batch Dependent Expiration Date: Batch dependent Amount: 10 mL Molecular Weight or Concentration: N/A Supplied as: Liquid Applications: a molecular tool for various biochemical applications Storage: 2-8 °C Keywords: Lipids Cholesterol Rich from adult bovine serum Grade: Biotechnology grade. All products are…

[meghanester]

Albumin Market Size, Growth with a CAGR of ~6% During 2023-2035 and Attain ~USD 10 Billion by 2035

Research Nester’s recent market research analysis on “Albumin Market: Global Demand Analysis & Opportunity Outlook 2035” delivers a detailed competitors analysis and a detailed overview of the global albumin market in terms of market segmentation by type, grade, application, and by region.

Rise in the Number of Heart Operation Performed in the World to Promote Global Market Share of Albumin

The global albumin market is estimated to grow majorly on account of the increasing prevalence of diseases, that affects human organs and requires the surgery. For instance, Patients undergoing cardiac surgery may incur severe blood loss, demanding the replacement of electrolytes and blood-related materials. Therefore, rise in the number of cardiac surgeries will drive the demand for albumins. Every year, roughly 2 million individuals worldwide receive open-heart surgery in order to treat various cardiovascular conditions. On the other hand, it is expected that in the coming few years, over 50 million adults in the world will be suffering from chronic liver disease, since it occurrence rate has increase from 4.5% to 9.5% in the world wide population. Albumin is employed as a component in the removal of excess fluid from the abdomen in liver illnesses.

Get a Sample PDF Brochure: https://www.researchnester.com/sample-request-5035

Besides this, various development has taken place in albumin, which makes it suitable for treating or aiding the treatment of a wide range of diseases. For instance, some researchers have suggested in COVID-19 patients if albumin serum is injected at low levels. It helps identify patients who are at high risk of developing pneumonia or respiratory failure and patients who require longer stays at the hospital. Furthermore, the rising popularity of plant-based albumin has significant drive the industry’s expansion. Plant-based albumins enable adaptation and change in function. As a result, scientists are attempting to create novel plant-based albumins through genetic engineering or by optimizing natural sources.

Some of the major growth factors and challenges that are associated with the growth of the global albumin market are:

Growth Drivers:

Surge in the Cardiovascular Surgeries

Rising Use of Albumins in the Research and Development of Drug Delivery Systems

Challenges:

Many people have shown allergic reactions towards albumin and there are strict regulatory policies for approving the use of albumin are some of the major factors anticipated to hamper the global market size of albumin. Individuals with known allergies to albumin or other proteins are more prone to experience allergic responses. The symptoms range from, skin rashes to serious effects, like breathlessness, chest pain, and others. On the other hand, there are many substitutes present in the market which is also expected to impede market growth. Albumins face competition from various alternative products and technologies in different applications. Some of the tough competitors are synthetic polymers, hydroxyethyl starch (HES), recombinant proteins, human serum-derived proteins, nanoparticles, and liposomes.

Request for customization @ https://www.researchnester.com/customized-reports-5035

On the basis of application, the global albumin market is segmented into drug formulation & vaccines, therapeutics, research. The therapeutics segment is anticipated to bring in the most revenue by the end of 2035, with a considerable CAGR over the forecast period. The growth of the segment is credited to growing incidents involving severe burns and other injuries. Albumin is utilized as a plasma expander to restore and maintain intravascular volumes, which is lost due to severe burns. Furthermore, based on type, the market is further fragmented into recombinant albumin, human serum albumin, bovine albumin. Out of all, human serum albumin has shown tremendous growth and will be growing notably in the forecast period. The rising spending on the research and development of new drugs has boosted the demand for human serum albumin,

Furthermore, by the end of 2035, Europe is expected to control a significant share of the market.  Improved understanding of the role of albumin in various conditions and the development of innovative therapies have increased the utilization of albumin in Europe. The advancements in medical technology and practices have expanded the application of albumins in healthcare. On the other hand, the rising prevalence of liver diseases, such as cirrhosis, hepatitis, and liver failure are also anticipated to boost the market growth in the region.

Obtain this Report @ https://www.researchnester.com/reports/albumin-market/5035

This report also provides the existing competitive scenario of some of the key players of the global albumin market which includes company profiling of Octapharma AG, Bio Med International Pvt. Ltd., Baxter, Apotex Inc., Novozymes, Mitsubishi Tanabe Pharma America Inc., Merck KGaA, Akron Biotech, Thermo Fisher Scientific, and Grifols, S.A

About Us

Research Nester is a leading service provider for strategic market research and consulting. We aim to provide unbiased, unparalleled market insights and industry analysis to help industries, conglomerates and executives to take wise decisions for their future marketing strategy, expansion and investment etc. We believe every business can expand to its new horizon, provided a right guidance at a right time is available through strategic minds. Our out of box thinking helps our clients to take wise decision in order to avoid future uncertainties.

Contact for more Info:

AJ Daniel

Email: info@researchnester.com

U.S. Phone: +1 646 586 9123

U.K. Phone: +44 203 608 5919

[nathanielaaron]

COPIED FROM ANOTHER PATRIOT.........

I gathered all vaccine ingredients into a list and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of that conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... But I'd have to easily say that they're all toxic to humans... Used in fertilizers... Pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... But you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... These are the ingredients used in vaccines... With binding agents to make sure the body won't flush these out... To keep the antibody levels up indefinitely...

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~  By  Iris Figueroa

Here are just SOME vaccine ingredients present in routine vaccines:

:black_medium_small_square:️Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

:black_medium_small_square:️Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

:black_medium_small_square:️Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

:black_medium_small_square:️Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

:black_medium_small_square:️Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

:black_medium_small_square:️Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

:black_medium_small_square:️Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

:black_medium_small_square:️Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

:black_medium_small_square:️Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

:black_medium_small_square:️African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

:black_medium_small_square:️Acetone - Can cause kidney, liver, and nerve damage.

:black_medium_small_square:️E.Coli - Yes, you read that right.

:black_medium_small_square:️DNA from porcine (pig) Circovirus type-1

:black_medium_small_square:️Human embryonic lung cell cultures (from aborted fetuses)

You can view all of these ingredients on the CDC website

[scatteredpearls]

“I gathered all vaccine ingredients into a list and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of that conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... But I'd have to easily say that they're all toxic to humans... Used in fertilizers... Pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... But you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... These are the ingredients used in vaccines... With binding agents to make sure the body won't flush these out... To keep the antibody levels up indefinitely...

He: WHAT?!

Your conclusion?

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~ By Iris Figueroa 🍃♥️🍃

✅ Detox & Cleanse * Educate *Support—->https://www.facebook.com/groups/1435942350040771/

✅INGREDIENTS TO VACCINES - You CANNOT make an educated decision without being educated.

Here are just SOME vaccine ingredients.

These are being INJECTED into your kids;

◾Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

◾Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

◾Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

◾Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

◾Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

◾Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

◾Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

◾Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

◾Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

◾African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

◾Acetone - Can cause kidney, liver, and nerve damage.

◾E.Coli - Yes, you read that right.

◾DNA from porcine (pig) Circovirus type-1

◾Human embryonic lung cell cultures (from aborted fetuses)

✳You can view all of these ingredients on the CDCs website. I encourage everyone to do their own research. Look up the MSDS on these chemicals. Read the thousands of peer reviewed studies that have evaluated the biological consequences these chemicals can have on the body, especially when being injected.

✳Fact check vaccine ingredients here:

https://www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table-2.pdf

#educatebeforeyouvaccinate #injectionisdifferentthaningestion #learntherisks #informedconsent

https://www.facebook.com/drryanberlin/posts/10160167051315215

Animal blood in vaccines

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1620411/pdf/amjphnation00376-0090.pdf

Food in vaccines, like peanut oil etc shows it causes food allergies

https://vaccinesafetycommission.org/pdfs/08-2015-Journal-Food-Allergies.pdf?fbclid=IwAR1_LCjNcaF36gVwoqz3k59njsc4ShWeJIi-cMZK3tt_VqydCL2vnB0AQdM

Vaccine Guide

https://vaccine.guide

Legislative informal hearing on mandatory vaccination.

https://www.facebook.com/100010598107344/videos/1007370642959530?d=n&sfns=mo

[petnews2day]

Intranasal delivery of a rationally attenuated SARS-CoV-2 is immunogenic and protective in Syrian hamsters

New Post has been published on https://petnews2day.com/pet-news/small-pet-news/intranasal-delivery-of-a-rationally-attenuated-sars-cov-2-is-immunogenic-and-protective-in-syrian-hamsters/

Intranasal delivery of a rationally attenuated SARS-CoV-2 is immunogenic and protective in Syrian hamsters

Research described here complies with all relevant ethical regulations and has been approved by the US Food and Drug Aministration Institutional Biosafety Committee. All critical reagents are listed in Supplementary Table 1.

Cells and viruses

Vero E6 cell line (Cat # CRL-1586) was purchased from American Type Cell Collection (ATCC) and cultured in Eagle’s minimal essential medium (MEM) supplemented with 10% fetal bovine serum (Invitrogen) and 1% penicillin/streptomycin and L-glutamine. A549-hACE2 (NR-53821) cells were obtained from BEI Resources and maintained in DMEM supplemented with 5% penicillin and streptomycin, and 10% fetal bovine serum (FBS) at 37 °C with 5% CO2. Lenti-X cell line was purchased from Takarabio (Cat No. 632180) and maintained in DMEM supplemented with 5% penicillin and streptomycin, and 10% fetal bovine serum (FBS) at 37 °C with 5% CO2.

EpiAirway cells (AIR-100-HCF) and culturing media were purchased from MatTek. EpiAirway is a ready-to-use, 3D mucociliary tissue model consisting of normal, human-derived tracheal/bronchial epithelial cells cultured at the air-liquid interface (ALI). Cells were cultured in MatTek proprietary media for 2 days prior to usage. Mucus was washed off at the time of infection.

The SARS-CoV-2 isolate WA1/2020 (NR-52281, lot 70033175) was obtained from BEI Resources, NIAID, NIH, and had been passed three times on Vero cells and 1 time on Vero E6 cells prior to acquisition. It was further passed once on Vero E6 cells in our lab. The virus has been sequenced and verified to contain no mutation to its original seed virus.

Production of SARS-CoV-2 recombinant virus

SARS-CoV-2 recombinant virus was generated using a 7-plasmid reverse genetic system which was based on the virus strain (2019-nCoV/USA_WA1/2020) isolated from the first reported SARS-CoV-2 case in the U.S.57. The initial 7 plasmids were generous gifts from Dr. P-Y Shi (UTMB). Upon receival, fragment 4 was subsequently subcloned into a low-copy plasmid pSMART LCAmp (Lucigen) to increase stability. To introduce Nsp1N128S/K129E and K164A/H165A mutations, pUC57-CoV2-F1 plasmids containing mutated Nsp1 were first created by using overlap PCR method with the following primers:

M13F: gtaaaacgacggccagt

N128S/K129Ef: taagaacggtAGTGAGggagctggtggccatagtta

N128S/K129E r: caccagctccCTCACTaccgttcttacgaagaagaa

K164A/H165Af: aaaactggaacactGCcGCcagcagtggtgttacccgtga

K164A/H165Ar: gggtaacaccactgctgGCgGCagtgttccagttttcttgaa

NheIr: cacgagcagcctctgatgca

PCR fragments were digested by Bgl II/Nhe I and ligated into Bgl II/Nhe I digested F1 plasmid. The spike ΔPRRA mutation was introduced in to pUC57-CoV2-F6 by using overlap PCR with primers:

M13F: gtaaaacgacggccagt

ΔPRRA-f: actcagactaattctcgtagtgtagctagtcaatc

ΔPRRA-r: actagctacactacgagaattagtctgagtctgat

BglIIr: cagcatctgcaagtgtcact

PCR fragments were digested by Kpn I/Bgl II and ligated into Kpn I/Bgl II digested F6 plasmid.

To delete the ORF6-ORF8 region, an overlap PCR was performed using the following primers:

Mf: ttaattttagccatggcaga

ORF68f: tttgcttgtacagtaaacgaacaaactaaaatgtc

ORF68r: ttttagtttgttcgtttactgtacaagcaaagcaa

AvrIIr: gaagtccagcttctggccca

PCR fragments were digested by Mlu I/Avr II and ligated into Mlu I/Avr II digested pCC1-CoV-2-F7 plasmid. The resulted plasmids were validated by restriction enzyme digestion and Sanger sequencing.

In vitro transcription and electroporation were carried following procedures that were detailed elsewhere58. To recover the virus, the RNA transcript was electroporated into Vero E6 cells. Virus after passage 1 was titrated by plaque forming assay in Vero E6 cells and verified by deep sequencing.

Hamster challenge experiments

Adult male outbred Syrian hamsters were previously purchased from Envigo and held at FDA vivarium. All experiments were performed within the biosafety level 3 (BSL-3) suite on the White Oak campus of the U.S. Food and Drug Administration. The animals were implanted subcutaneously with IPTT-300 transponders (BMDS), randomized, and housed 2 per cage in sealed, individually ventilated rat cages (Allentown). Hamsters were fed irradiated 5P76 (Lab Diet) ad lib, housed on autoclaved aspen chip bedding with reverse osmosis-treated water provided in bottles, and all animals were acclimatized at the BSL3 facility for 4–6 days or more prior to the experiments. The study protocol details were approved by the White Oak Consolidated Animal Care and Use Committee and carried out in accordance with the PHS Policy on Humane Care & Use of Laboratory Animals.

For challenge studies, adult (6–12 months old) Syrian hamsters were anesthetized with 3–5% isoflurane following procedures as described previously38,59. Intranasal inoculation was done by pipetting 104 PFU or desirable doses of SARS-CoV-2 in 50 µl volume dropwise into the nostrils of the hamster under anesthesia. Following infection, hamsters were monitored daily for clinical signs and weight loss. Nasal wash samples taken on days 1-, 2-, 3-, and 4-day post infection to test for sgRNA by RT-qPCR and infectious virus by TCID50 in Vero E6 cells. Nasal washes were collected by pipetting ~160 µl sterile phosphate-buffered saline into one nostril when hamsters were anesthetized by 3–5% isoflurane. For tissue collection, a subset of hamsters was humanely euthanized by intraperitoneal injection of pentobarbital at 200 mg/kg and lungs for histopathology. Blood collection was performed under anesthesia (3–5% isoflurane) through gingival vein puncture or cardiac puncture when animals were euthanized.

Mouse infection experiments

Female adult K18-hACE2 mice (12 weeks ago) were previously purchased from the Jackson laboratory and held at FDA vivarium. All experiments were performed within the biosafety level 3 (BSL-3) suite on the White Oak campus of the U.S. Food and Drug Administration. The study protocol details were approved by the White Oak Consolidated Animal Care and Use Committee and carried out in accordance with the PHS Policy on Humane Care & Use of Laboratory Animals.

For infection studies, mice were first anesthetized by 3–5% isoflurane. Intranasal inoculation was done by pipetting 105 PFU SARS-CoV-2 in 50 µl volume dropwise into the nostrils of the mouse. Mice were weighed and observed daily. For tissue collections, mice were euthanized by CO2 overdose on days 2, 4, 6 as necessary.

SARS-CoV-2 pseudovirus production and neutralization assay

Procedures as described previously38,59. In brief, 50 μL of SARS-CoV-2 S pseudovirions were pre-incubated with an equal volume of medium containing serum at varying dilutions at room temperature for 1 h, then virus-antibody mixtures were added to Vero E6 cells in a 96-well plate. After a 3 h incubation, the inoculum was replaced with fresh medium. Cells were lysed 48 h later, and luciferase activity was measured using luciferin-containing substrate. Controls included cell only control, virus without any antibody control, and positive control sera. The end-point titers were calculated as the last serum dilution resulting in at least 50% SARS-CoV-2 neutralization. A NIBSC anti-SARS-CoV-2 antibody (20/130) was included as a positive control.

RNA isolation and qRT-PCR

Procedures as described previously38,59. In brief, RNA was extracted from 50 μl NW or 0.1 g tissue homogenates using QIAamp vRNA mini kit or the RNeasy 96 kit (QIAGEN) and eluted with 60 μl of water. 5 μL RNA was used for each reaction in real-time RT-PCR. When graphing the results in Prism 8, values below the limit of quantification (LoQ) were arbitrarily set to half of the LoQ values. Unless otherwise specified, the unit for RNA copies are as presented as Log10 RNA copies/5 μl nasal wash or Log10 RNA copes/0.1 g tissue homogenates,

RNAseq

To prepare sequencing libraries, RNA was first extracted using the Trizol-chloroform method from the lung homogenates and nasal turbinates. The aqueous portion was further purified using RNeasy mini kit (Qiagen, Gaithersburg, MD). RNA quality was assessed using Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA), and the RNA integration numbers (RIN) were all greater than 9. An aliquot (1 μg) of each sample of total RNA was used to prepare sequencing libraries using Illumina Stranded Messenger RNA Prep (ligation based). The cDNA libraries were normalized and loaded onto a NovaSeq 6000 sequencer (Illumina, San Diego, CA) for deep sequencing of paired-end reads of 2 × 100 cycles. The sequencing reads for each sample were mapped to the respective reference genomes of Mesocricetus auratus (BCM_Maur_2.0) by Tophat (v2.1.2). Cufflinks (v2.2.1) was then used to assemble transcripts, estimate abundances and test for differential expression. The sequencing and initial data analysis using Qiagen CLC Genomics Workbench (version 21) was performed by FDA Next Generation Sequencing Core Facility. The sequencing and initial data analysis using Qiagen CLC Genomics Workbench (version 21) was performed by FDA Next Generation Sequencing Core Facility. Raw and processed data have been deposited to NCBI (GEO accession number GSE199922s).

Further data analysis was done using R Studio 1.4.1106 (http://www.R-project.org). Heatmaps were constructed using heatmap library. The gene list for signaling pathways was obtained from hallmark gene sets in Molecular Signatures Database (MSigDB)60. The figures were assembled in Adobe Photoshop. This work utilized the computational resources of the NIH HPC Biowulf cluster (http://hpc.nih.gov).

Histopathology analyses

Procedures as described previously38,59. Tissues (hearts, brains, lungs, trachea, and nasal turbinates) were fixed in 10% neural buffered formalin overnight and then processed for paraffin embedding. The 5-μm sections were stained with hematoxylin and eosin for histopathological examinations. Images were scanned using an Aperio ImageScope. For hamster tissues, blinded samples were graded by a licensed pathologist for the following twelve categories: consolidation, alveolar wall thickening, alveolar airway infiltrates, perivascular infiltrates, perivascular edema, type II pneumocyte hyperplasia, atypical pneumocyte hyperplasia, bronchiole mucosal hyperplasia, bronchiole airway infiltrates, proteinaceous fluid, hemorrhage, vasculitis. Grading: 0 = none, 1 = mild, 2 = moderate, 3 = severe. A graph was prepared by summing up the score in each category. Mouse tissues were scored based on the following categories: consolidation, alveolar wall thickening, alveolar airway infiltrates, perivascular Infiltrates, perivascular edema, peribronchiolar infiltrates, type II pneumocyte hyperplasia, necrosis (alveoli and bronchiole), bronchiole mucosal hyperplasia, bronchiole airway infiltrates, proteinaceous fluid, hemorrhage, vasculitis. Grading scale: 0 = none, 1 = mild, 2 = moderate, 3 = severe.

In situ hybridization (RNAscope)

To detect SARS-CoV-2 genomic RNA in FFPE tissues, ISH was performed using the RNAscope 2.5 HD RED kit, a single plex assay method (Advanced Cell Diagnostics; Catalog 322373) according to the manufacturer’s instructions. Briefly, Mm PPIB probe detecting peptidylprolyl isomerase B gene (housekeeping gene) (catalog 313911, positive-control RNA probe), dapB probe detecting dihydrodipicolinate reductase gene from Bacillus subtilis strain SMY (a soil bacterium) (catalog 310043, negative-control RNA probe) and V-nCoV2019-orf1ab (catalog 895661) targeting SARS-CoV-2 positive-sense (genomic) RNA. Tissue sections were deparaffinized with xylene, underwent a series of ethanol washes and peroxidase blocking, and were then heated in kit-provided antigen retrieval buffer and digested by kit-provided proteinase. Sections were exposed to ISH target probes and incubated at 40 °C in a hybridization oven for 2 h. After rinsing, ISH signal was amplified using kit-provided pre-amplifier and amplifier conjugated to alkaline phosphatase and incubated with a fast-red substrate solution for 10 min at room temperature. Sections were then stained with 50% hematoxylin solution followed by 0.02% ammonium water treatment, dried in a 60 °C dry oven, mounted, and stored at 4 °C until image analysis.

Lung immunofluorescence analyses

Formalin-fixed paraffin-embedded (FFPE) lung sections 4 µm thick were dewaxed, rehydrated, and heat-treated in a microwave oven for 15 min in 10 mM Tris/1 mM EDTA buffer (pH 9.0). After cooling for 30 min at room temperature, heat-retrieved sections were blocked in PBST with 2.5% bovine serum albumin (BSA) for 30 min at RT followed by overnight incubation at 4 °C with primary antibodies in 1% BSA. Primary antibodies used included SARS nucleocapsid protein (1:800, Sino Biologicals, 40143-MM05), prosurfactant protein C (1:200, EMD Millipore, AB3786), Iba1 (1:100, Abcam, ab5076), and RAGE (1:400, Abcam, ab216329). Sections were rinsed and incubated with 1:500 secondary antibodies congujated with Alexa Fluor 488 (A-21206) and Alexa Fluor 647 (A-31571, A-21447) for 1 h at RT (ThermoFisher, Waltham, MA). Nuclei were counterstained with Hoechst 33342. For double labeling experiments, primary antibodies were mixed and incubated overnight at 4 °C. For negative controls, sections were incubated without the primary antibody or mouse and rabbit isotype antibody controls. Sections stained with conjugated secondary antibodies alone showed no specific staining. Whole-slide fluorescence imaging was performed using a Hamamatsu NanoZoomer 2.0-RS whole-slide digital scanner equipped with a ×20 objective and a fluorescence module #L11600. Analysis software NDP.view2 was used for image processing (Hamamatsu Photonics, Japan).

TCID50

Procedures as described previously38,59. In brief, Vero E6 cells were plated the day before infection into 96-well plates at 1.5 × 104 cells/well. On the day of the experiment, serial dilutions of 20 μl nasal wash samples were made in media and a total of six to eight wells were infected with each serial dilution of the virus. After 48 h incubation, cells were fixed in 4% PFA followed by staining with 0.1% crystal violet. The TCID50 was then calculated using the formula: log(TCID50) = log(do) + log(R) (f + 1). Where do represents the dilution giving a positive well, f is a number derived from the number of positive wells calculated by a moving average, and R is the dilution factor.

Plaque assay

Nasal wash samples were 10-fold serially diluted and added to a 24-well plate containing freshly confluent with Vero E6 cells. For tissue samples, entire trachea or nasal turbinates or the left lobe of the lung (~0.2 g) were resuspended in 1 milliliter MEM and homogenized on a Precellys Evolution tissue homogenizer with a Cooling Unit (Bertin). Tissue homogenates were then 10-fold serially diluted and added to Vero E6. After 1 h the mixture was removed and replenished with Tragacanth gum overlay (final concentration 0.3%). Cells were incubated at 37 °C and 5% CO2 for 2 days, then fixed with 4% paraformaldehyde, followed by staining of cells with 0.1% crystal violet in 20% methanol for 5–10 min. The infectious titers were then calculated and plotted as plaque forming units per milliliter (PFU/ml).

Focus-forming assay

ALI cell culture supernatants were 10-fold serially diluted in 96-well plates and dilutions added to 96-well black-well plates for fluorescent focus-forming assays in H1299-hACE2 cells. After 1 h the Tragacanth gum overlay (final concentration 0.3%) was added. Cells were incubated at 37 °C and 5% CO2 for 1 day, then fixed with 4% paraformaldehyde, followed by staining of cells with primary rabbit anti-nucleocapsid Wuhan-1 antibody (custom made by Genscript) overnight followed by secondary anti-rabbit Alexa-488 conjugated antibody and DAPI staining. The infectious titers were then counted using Gen5 software on a Cytation7 machine and calculated and plotted as focus-forming units per milliliter (FFU/ml).

Measurement of antibody by ELISA

SARS-CoV-2 S and RBD antigens for ELISA were prepared in a baculovirus expression system using procedures as published elsewhere61. Briefly, Immunlon 2HB plates were coated with recombinant S or RBD protein at 1 µg/mL overnight at 4 °C. Test serum samples were pre-diluted in assay diluent (PBS containing 0.05% Tween-20 [PBST] and 10% fetal bovine serum), followed by serial two-fold dilutions of each sample in duplicates across the plate. Plates were incubated with the test serum samples for 2 h at 37 °C. After rigorous plate washes in a microplate washer, a secondary antibody (anti-hamster IgG) conjugated to HRP (6060-05, SouthernBiotech, Birmingham, AL) was added to wells at 1:4000 dilution (hamster IgG ELISA). Plates were incubated with secondary antibody for 1 h, washed, and ABTS/H2O2 peroxidase substrate (SeraCare, Gaithersburg, MD) was added to assay wells. After 20 to 30 min at ambient temperature, reactions were stopped with 1% SDS, and OD405 values were captured on the Versamax microplate reader with the Softmax Pro 7 software installed (Molecular Devices, San Jose, CA). The assay endpoint was a mean OD405 of 0.05 for duplicate wells for the full-length S ELISA and a mean OD405 of 0.01 for the RBD ELISA. The reciprocal of the highest serum dilution at which the mean OD405 value averaged ≥0.05 (full-length S ELISA) or ≥0.01 (RBD ELISA) was the IgG titer. To determine the serum IgA antibody titers, each test serum sample was diluted from 1:160. The sandwich Rabbit anti-hamster IgA antibody (sab 3001a, Brookwood Biomedical, Jemison, Alabama) was added at 1:4000 dilution followed by 1:4000 Goat anti-rabbit-HRP (4030-05, Southern Biotech, Birmingham, AL).

Statistical analysis

One-way ANOVA was used to calculate statistical significance through GraphPad Prism (9.1.2) software for Windows, GraphPad Software, San Diego, California, USA.

Reporting summary

Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article.

[slowlyteenagestarlight]

Dig in… share no credit or acknowledgment required. People have no clue about what they’re acquiescence with.

Don't put this on Facebook unless you want a ban.

Read this!!! 😳

Someone's BRILLIANT HOMEWORK!!!!!

“I made a list of all vaccine ingredients and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of the conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... but I'd have to say easily that they're all toxic to humans... Used in fertilizers, pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... but you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... these are the ingredients used in vaccines... with binding agents to make sure the body won't flush them out... to keep the antibody levels up indefinitely...

He: WHAT?!

Your conclusion?

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~ By Iris Figueroa 🍃♥️🍃

✅INGREDIENTS IN VACCINES - You CANNOT make an educated decision without being educated.

Here are just SOME vaccine ingredients.

These are being INJECTED into your kids;

◾️Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

◾️Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

◾️Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

◾️Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

◾️Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

◾️Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

◾️Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

◾️Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

◾️Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

◾️African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

◾️Acetone - Can cause kidney, liver, and nerve damage.

◾️E.Coli - Yes, you read that right.

◾️DNA from porcine (pig) Circovirus type-1

◾️Human embryonic lung cell cultures (from aborted fetuses)

✳️You can view all of these ingredients on the CDCs website. I encourage everyone to do their own research. Look up the MSDS on these chemicals. Read the thousands of peer reviewed studies that have evaluated the biological consequences these chemicals can have on the body, especially when being injected.

✳️Fact check vaccine ingredients here:

https://www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table-2.pdf

PLEASE SHARE!!!

www.cdc.gov

cdc.govDon't put this on Facebook unless you want a ban.

Read this!!! 😳

Sent from my iPhone

[awesomecooperlove]

“I gathered all vaccine ingredients into a list and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of that conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... But I'd have to easily say that they're all toxic to humans... Used in fertilizers... Pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... But you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... These are the ingredients used in vaccines... With binding agents to make sure the body won't flush these out... To keep the antibody levels up indefinitely...

He: WHAT?!

Your conclusion?

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~ By Iris Figueroa 🍃♥️🍃

✅INGREDIENTS TO VACCINES - You CANNOT make an educated decision without being educated.

Here are just SOME vaccine ingredients.

These are being INJECTED into your kids;

◾️Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

◾️Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

◾️Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

◾️Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

◾️Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

◾️Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

◾️Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

◾️Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

◾️Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

◾️African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

◾️Acetone - Can cause kidney, liver, and nerve damage.

◾️E.Coli - Yes, you read that right.

◾️DNA from porcine (pig) Circovirus type-1

◾️Human embryonic lung cell cultures (from aborted fetuses)

✳️You can view all of these ingredients on the CDCs website. I encourage everyone to do their own research. Look up the MSDS on these chemicals. Read the thousands of peer reviewed studies that have evaluated the biological consequences these chemicals can have on the body, especially when being injected.

✳️Fact check vaccine ingredients here:

https://www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table-2.pdf

[jesse-pinkman123]

Cell Culture Media for Vaccine Market Forecast Opportunity Analysis - 2027

Global Cell Culture Media for Vaccine Market, by Media Type (Bovine Derived, Porcine Derived, and Animal Free/Serum Free Cell Culture), by Application (Human Vaccine and Animal Vaccine), and by Region (North America, Latin America, Europe, Asia Pacific, Middle East, and Africa) was valued at US$ 1,235.5 million in 2017 and is projected to exhibit a CAGR of 7.7% over the forecast period (2018 – 2026).

Key players in the market are focused on research and development in order to develop novel research grade products to cater the need of researchers. For instance, in November 2017, Proliant Biologicals launched New Zealand manufactured Reagent Grade (Fatty Acid-Free) BSA. The product is derived from New Zealand-origin healthy animals. Moreover, increasing number of government initiatives to prevent certain diseases and epidemic caused due to vaccine-preventable diseases is expected to boost the market growth. Furthermore, increasing number of countries are conducting national immunization programs to provide vaccination and promote awareness regarding advantages of vaccination to fight fatal diseases with natural immunity among adults as well as children. For instance, in 2015, the Centers for Disease Control and Prevention (CDC), the Food and Drug Administration (FDA), and the National Institutes of Health (NIH) partnered with the U.S. Department of Health and Human Services Biomedical Advanced Research and Development Authority (BARDA) to support the advanced development of new and better influenza vaccines as a part of inter-agency government effort in the U.S.

* The sample copy includes: Report Summary, Table of Contents, Segmentation, Competitive Landscape, Report Structure, Methodology.

Request a sample copy of this report: https://www.coherentmarketinsights.com/insight/request-sample/2608

Browse 33 Market Data Tables and 27 Figures spread through 197 Pages and in-depth TOC on ‘Cell Culture Media for Vaccine Market’- Global Forecast to 2026, by Media Type (Bovine Derived, Porcine Derived, and Animal Free/Serum Free Cell Culture), by Application (Human Vaccine and Animal Vaccine), and by Region (North America, Latin America, Europe, Asia Pacific, Middle East, and Africa)

Key players in the market are focused on adopting strategies such as mergers and acquisitions to enhance their product portfolio. For instance, In November 2015, Merck KGaA, which is a parent company of Merck Millipore, acquired the U.S.-based life science company, Sigma-Aldrich Corporation, at US$ 17 Bn through its U.S. subsidiary EMD Millipore. A combined entity known as MilliporeSigma was formed in the U.S. market and this acquisition made Merck Millipore one of the top three players in the global life science market.

Key Takeaways of the Global Cell Culture Media for Vaccine Market:

The global cell culture media for vaccine market is expected to exhibit a CAGR of 7.7 % over the forecast period, owing to increasing prevalence of infectious diseases, increasing number of government initiatives, and increasing availability of innovative bovine blood plasma derivatives. For instance, in August 2017, Animal Free Research UK and the 3Rs-Centre Utrecht Life Sciences launched the Fetal Calf Serum (FCS)-free database. The database enables researchers to identify serum-free media for specific cell types, thereby contributing to the adoption of animal free media for research.

Browse Research Report: https://www.coherentmarketinsights.com/market-insight/cell-culture-media-for-vaccine-market-2608

Among application type, human vaccine segment is estimated to hold a dominant position in the cell culture media for vaccine market in 2018, owing to higher usage of bovine derived serum products in biotechnology research, vaccine production, cloning, and in vitro fertilization

Among media type, bovine derived is estimated to hold a dominant position in the cell culture media for vaccine market in 2018, as manufacturing of viral vaccines require fetal bovine serum or other sera as a source of nutrition or growth media for the virus growth

Key players operating in the global cell culture media for vaccine market include, HiMedia Laboratories, Thermo Fisher Scientific Inc., Merck KGaA, General Electric Company, Sartorius AG, Proliant, Inc., Bovogen Biologicals Pty. Ltd., Rocky Mountain Biologicals, Valley Biomedical, Moregate BioTech, Atlanta Biologicals, Creative-Biolabs, Life Technologies (India) Pvt Ltd., Axil Scientific Pte Ltd., Indian Immunologicals Limited, Serum Institute of India Pvt. Ltd., Sisco Research Laboratories Pvt. Ltd., and Valneva SE

Buy-Now this research report: https://www.coherentmarketinsights.com/insight/buy-now/2608

About Coherent Market Insights:            

Coherent Market Insights is a prominent market research and consulting firm offering action-ready syndicated research reports, custom market analysis, consulting services, and competitive analysis through various recommendations related to emerging market trends, technologies, and potential absolute dollar opportunity.

Contact Us:

mailto:sales@coherentmarketinsights.com

U.S. Office:

Name:  Mr. Shah

Coherent Market Insights 1001 4th Ave,

# 3200 Seattle, WA 98154, U.S.

US :  +1-206-701-6702

UK :  +44-020-8133-4027

JAPAN :  +050-5539-1737

[the-other-news]

“I gathered all vaccine ingredients into a list and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of that conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... But I'd have to easily say that they're all toxic to humans... Used in fertilizers... Pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... But you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... These are the ingredients used in vaccines... With binding agents to make sure the body won't flush these out... To keep the antibody levels up indefinitely...

He: WHAT?!

Your conclusion?

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~ By I Figueroa 🍃♥️🍃

✅INGREDIENTS TO VACCINES - You CANNOT make an educated decision without being educated.

Here are just SOME vaccine ingredients.

These are being INJECTED into your kids;

◾️Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

◾️Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

◾️Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

◾️Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

◾️Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

◾️Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

◾️Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

◾️Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

◾️Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

◾️African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

◾️Acetone - Can cause kidney, liver, and nerve damage.

◾️E.Coli - Yes, you read that right.

◾️DNA from porcine (pig) Circovirus type-1

◾️Human embryonic lung cell cultures (from aborted fetuses)

✳️You can view all of these ingredients on the CDCs website. I encourage everyone to do their own research. Look up the MSDS on these chemicals. Read the thousands of peer reviewed studies that have evaluated the biological consequences these chemicals can have on the body, especially when being injected.

✳️Fact check vaccine ingredients here:https://www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table-2.pdf

Source: World Doctors Alliance

[athiranair23]

Cell Culture Media for Vaccine Market Analysis (2020-2027)

Cell culture media consists an appropriate source of energy and compounds, which supports growth of cells and regulate the cell cycle. A typical culture medium is composed of vitamins, inorganic salts, complement of amino acids, glucose, and serum which acts as source of growth factors, hormones, and attachment factors. Furthermore, the medium also helps to maintain pH and osmolality.

Market Dynamics

Increasing prevalence of infectious diseases is a major factor driving the global cell culture media for vaccine market growth. According to the Centers for Disease Control and Prevention (CDC), in August 2017, around 104 laboratory-confirmed influenza-associated pediatric deaths were reported, where 66 cases were associated with influenza A viruses and 38 cases were associated with influenza B viruses in the U.S. Moreover, the government initiatives for increasing awareness among the populace regarding prevention of diseases facilitated by vaccines is expected to boost the market growth. For instance, according to the Centers for Disease Control and Prevention (CDC), in 2015, flu vaccinations prevented around 1.9 million flu illnesses, 966,000 flu-associated medical visits, and around 67,000 flu-associated hospitalizations, representing 6.5% lowering of disease burden in the U.S.

However, high risk of contamination may hinder the market growth. According to the article published in ResearchGate, 2014, fetal bovine serum (FBS) is known to contain factors such as endotoxins, mycoplasma, viral contaminants, and prions that can lead to bovine spongiform encephalopathy (BSE).

Key features of the study:

·         This report provides in-depth analysis of the global cell culture media for vaccine market and provides market size (US$ million) and Compound Annual Growth Rate (CAGR %) for the forecast period (2018 – 2026), considering 2017 as the base year

·         It elucidates potential revenue opportunities across different segments and explains attractive investment proposition matrices for this market

·         This study also provides key insights into market drivers, restraints, opportunities, impact analysis, Porter’s analysis, PEST analysis, regulatory scenario, key developments, raw material analysis, and supply chain analysis

·         It profiles key players in the global cell culture media for vaccine market based on the following parameters – company overview, financial performance, product portfolio, market presence, distribution strategies, key developments, strategies, and future plans

·         Key companies covered as a part of this study include HiMedia Laboratories, Thermo Fisher Scientific Inc., Merck KGaA, General Electric Company, Sartorius AG, Proliant, Inc., Bovogen Biologicals Pty. Ltd., Rocky Mountain Biologicals, Valley Biomedical, Moregate BioTech, Atlanta Biologicals, Creative-Biolabs, Life Technologies (India) Pvt Ltd., Axil Scientific Pte Ltd., Indian Immunologicals Limited, Serum Institute of India Pvt. Ltd., Sisco Research Laboratories Pvt. Ltd., and Valneva SE

·         Insights from this report would allow marketers and the management authorities of the companies to make informed decisions regarding their future product launches, technology up-gradation, market expansion, and marketing tactics

·         The global cell culture media for vaccine market report caters to various stakeholders in this industry including investors, suppliers, product manufacturers, distributors, new entrants, and financial analysts

·         Stakeholders would have ease in decision-making through various strategy matrices used in analyzing the global cell culture media for vaccine market

Detailed Segmentation:

·         Global Cell Culture Media for Vaccine Market, By Media Type:

o    Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

o    Porcine Derived

o    Animal free/Serum Free Cell Culture

·         Global Cell Culture Media for Vaccine Market, By Application:

o    Human Vaccines

o    Animal Vaccines

·         Global Cell Culture Media for Vaccine Market, By Geography:

o    North America

§  By Media Type:

§  Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

§  Porcine Derived

§  Animal free/Serum Free Cell Culture

§  By Application:

§  Human Vaccines

§  Animal Vaccines

§  By Country:

§  U.S.

§  Canada

o    Latin America

§  By Media Type:

§  Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

§  Porcine Derived

§  Animal free/Serum Free Cell Culture

§  By Application:

§  Human Vaccines

§  Animal Vaccines

§  By Country:

§  Brazil

§  Mexico

§  Argentina

§  Rest of Latin America

o    Europe

§  By Media Type:

§  Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

§  Porcine Derived

§  Animal free/Serum Free Cell Culture

§  By Application:

§  Human Vaccines

§  Animal Vaccines

§  By Country:

§  Germany

§  U.K.

§  France

§  Italy

§  Spain

§  Russia

§  Rest of Europe

o    Asia Pacific

§  By Media Type:

§  Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

§  Porcine Derived

§  Animal free/Serum Free Cell Culture

§  By Application:

§  Human Vaccines

§  Animal Vaccines

§  By Country:

§  China

§  India

§  Japan

§  Australia

§  South Korea

§  ASEAN

§  Rest of Asia Pacific

o    Middle East

§  By Media Type:

§  Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

§  Porcine Derived

§  Animal free/Serum Free Cell Culture

§  By Application:

§  Human Vaccines

§  Animal Vaccines

§  By Country:

§  GCC

§  Israel

§  Rest of Middle East

o    Africa

§  By Media Type:

§  Bovine Derived

§  Fetal bovine serum (FBS)

§  Newborn Calf Serum (NBCS)

§  Adult Bovine Serum (ABS)

§  Bovine Serum Albumin (BSA)

§  Bovine Calf Serum (BCS)

§  Porcine Derived

§  Animal free/Serum Free Cell Culture

§  By Application:

§  Human Vaccines

§  Animal Vaccines

§  By Country:

§  South Africa

§  Central Africa

§  North Africa

·         Company Profiles

o    HiMedia Laboratories *

§  Company Overview

§  Product Portfolio

§  Financial Performance

§  Key Highlights

§  Market Strategies

o    Thermo Fisher Scientific, Inc.

o    Merck KGaA

o    General Electric Company

o    Sartorius AG

o    Proliant, Inc.

o    Bovogen Biologicals Pty. Ltd.

o    Rocky Mountain Biologicals

o    Valley Biomedical

o    Moregate BioTech

o    Atlanta Biologicals

o    Creative-Biolabs

o    Life Technologies (India) Pvt Ltd.

o    Axil Scientific Pte Ltd.

o    Indian Immunologicals Limited

o    Serum Institute of India Pvt. Ltd.

o    Sisco Research Laboratories Pvt. Ltd.

o    Valneva SE

 “*” marked represents similar segmentation in other categories in the respective section.

  Request sample copy here :  

https://www.coherentmarketinsights.com/insight/request-sample/2608

Request PDF brochure here:        

https://www.coherentmarketinsights.com/insight/request-pdf/2608

Click here to buy: https://www.coherentmarketinsights.com/insight/buy-now/2608

CMI Services: https://www.coherentmarketinsights.com/services

About Us:

Coherent Market Insights is a global market intelligence and consulting organization focused on assisting our plethora of clients achieve transformational growth by helping them make critical business decisions.

What we provide:

·        Customized Market Research Services

·        Industry Analysis Services

·        Business Consulting Services

·        Market Intelligence Services

·        Long term Engagement Model

·        Country Specific Analysis

Contact Us:   

Mr. Shah

Coherent Market Insights Pvt. Ltd.

Address: 1001 4th ave, #3200 Seattle, WA 98154, U.S.

Phone: +1-206-701-6702

Email : sales@coherentmarketinsights.com

 Reference/Source: https://www.coherentmarketinsights.com/market-insight/cell-culture-media-for-vaccine-market-2608

[cleverninjaheart]

Goat Elisa Kits

pilot study is the first attempt in the country to estimate sero-prevalence of disease Bovine Johne's (BJD) with a screening of cattle and buffaloes which represents a large population of farmers' and livestock farming in house channels (Uttar Pradesh (UP) and Punjab) from cows Hariana and Murrah buffaloes in North India. 

Custom and in-house plate ELISA kit (using the protoplasm of the native antigen of Mycobacterium avium subsp. Paratuberculosis 'type Bison' strain of goat origin), originally developed for goats and sheep are standard in bovines and used for screening. For this study, 33 villages of southern and western UP were randomly selected and surveyed from 2001 to 2003. 

There were 7943 38 251 farming families have livestock, including cattle, buffalo, sheep and goats (per family 4.8% of livestock). Numerical, buffalo and cattle were 54.7% and 22.1%, respectively. 726 serum samples were collected from animals (4.2% of the 16 981 cattle with 4,375 family farmers) located in 33 villages were randomly surveyed. Serum samples (699), submitted to the Epidemiology Department of Veterinary College (Punjab Agricultural University, Ludhiana), in 2004 by farmers and organized agriculture livestock (buffaloes, 372, Cow, 327), screened by this ELISA kit. protoplasm soluble antigen prepared from Peta (S 5) 'Bison type' strains isolated from Goat Elisa Kits severely ill with Johne's disease. 

From a total of 1,425 cattle (buffalo and cow) serum samples were screened using ELISA kit custom, sero-prevalence of Johne's disease was 29.0% (28.6% at 29.8% on a buffalo and cattle) in North India. State-wise sero-prevalence was 31.9% and 23.3% in UP and Punjab, respectively. 

In UP, of 601 randomly sampled buffalo, sero-prevalence was 40.3% (16.6% in young adults and 40.9%) and 25.5% (10.5% in young adults and 26 3%) in the southern and western UP, respectively. Of the 125 animals screened, sero-prevalence was 42.6% (zero in young adults and 44.4%) and 30.0% (zero in young adults and 30.6%) in the southern and western UP, respectively respectively. Of the 699 serum samples screened from Ludhiana, Punjab, sero-prevalence of BJD was 23.0%. Sero-prevalence was 23.3% (12.1% and 24.4% in young adults) and 26.9% (27.2% and 26.8% in young adults) in buffalo and cattle, each. 

High prevalence of BJD in the original channel Murrah buffalo breed, and breed cattle Hariana correlated with poor productivity per animal and BJD may be the main cause. Indigenous ELISA kit is a rapid, economic and sensitive test for large-scale screening of buffalo and cattle population against the BJD can be cured.

[un-enfant-immature]

Bit Bio’s “enter button for the keyboard to the software of life” nabs the company $41.5 million

Bit Bio, the new startup which pitches itself as the “enter button for the keyboard to the software of life” only needed three weeks to raise its latest $41.5 million round of funding.

Originally known as Elpis Biotechnology and named for the Greek goddess of hope, the Cambridge, England-based company was founded by Mark Kotter in 2016 to commercialize technology that can reduce the cost and increase the production capacity for human cell lines. These cells can be used in targeted gene therapies and as a method to accelerate drug discovery at pharmaceutical companies.

The company’s goal is to be able to reproduce every human cell type.

“We’re just at a very crucial time in biology and medicine and the bottleneck that has become really clear is a scalable source of robust human cells,” said Kotter. “For drug discovery this is important. When you look at failure rates in clinical trials they’re at an all time high… that’s in direct contradiction to the massive advancements in biotechnology in research and the field.”

In the seventeen years since scientists completely mapped the human genome, and eight years since scientists began using the gene editing technology known as CRISPR to edit genetic material, there’s been an explosion of treatments based on individual patient’s genetic material and new drugs developed to more precisely target the mechanisms that pathogens use to spread through organisms.

These treatments and the small molecule drugs being created to stop the spread of pathogens or reduce the effects of disease require significant testing before coming to market — and Bit Bio’s founder thinks his company can both reduce the time to market and offer new treatments for patients.

It’s a thesis that had investors like the famous serial biotech entrepreneur, Richard Klausner, who served as the former director of the National Cancer Institute and founder of revolutionary biotech companies like Lyell Immunopharma, Juno, and Grail, leaping at the chance to invest in Bit Bio’s business, according to Kotter.

Joining Klausner are the famous biotech investment firms Foresite Capital, Blueyard Capital and Arch Venture Partners.

“Bit Bio is based on beautiful science. The company’s technology has the potential to bring the long-awaited precision and reliability of engineering to the application of stem cells,” said Klausner in a statement. “Bit Bio’s approach represents a paradigm shift in biology that will enable a new generation of cell therapies, improving the lives of millions.”

Photo: Andrew Brookes/Getty Images

Kotter’s own path to develop the technology which lies at the heart of Bit Bio’s business began a decade ago in a laboratory in Cambridge University. It was there that he began research building on the revolutionary discoveries of Shinya Yamanaka, which enabled scientists to transform human adult cells into embryonic stem cells.

“What we did is what Yamanaka did. We turned everything upside down. We want to know how each cell is defined… and once we know that we can flip the switch,” said Kotter. “We find out which transcription factors code for a single cell and we turn it on.”

Kotter said the technology is like uploading a new program into the embryonic stem cell.

Although the company is still in its early days, it has managed to attract a few key customers and launch a sister company based on the technology. That company, Meatable, is using the same process to make lab-grown pork.

Dutch startup Meatable is developing lab-grown pork and has $10 million in new financing to do it

Meatable is the earliest claimant to a commercially viable, patented process for manufacturing meat cells without the need to kill an animal as a prerequisite for cell differentiation and growth.

Other companies have relied on fetal bovine serum or Chinese hamster ovaries to stimulate cell division and production, but Meatable says it has developed a process where it can sample tissue from an animal, revert that tissue to a pluripotent stem cell, then culture that cell sample into muscle and fat to produce the pork products that palates around the world crave.

“We know which DNA sequence is responsible for moving an early-stage cell to a muscle cell,” says Meatable chief executive Krijn De Nood.

If that sounds similar to Bit Bio, that’s because it’s the same tech — just used to make animal instead of human cells.

Image: PASIEKA/SCIENCE PHOTO LIBRARY/Getty Images

If Meatable is one way to commercialize the cell differentiation technology, Bit Bio’s partnership with the drug development company Charles River Laboratories is another.

“We actually do have a revenue generating business side using human cells for research and drug discovery. We have a partnership with Charles River Laboratories the large preclinical contract research organization,” Kotter said. “That partnership is where we have given early access to our technology to Charles River… They have their own usual business clients who want them to help with their drug discovery. The big bottleneck at the moment is access to human cells.”

Drug trials fail because the treatments developed either are toxic or don’t work in humans. The difference is that most experiments to prove how effective the treatments are rely on animal testing before making the leap to human trials, Kotter said.

The company is also preparing to develop its own cell therapies, according to Kotter. There, the biggest selling point is the increased precision that  Bit Bio can bring to precision medicine, said Kotter. “If you look at these cell therapies at the moment you get mixed bags of cells. There are some that work and some that have dangerous side effects. We think we can be precise [and] safety is the biggest thing at this point.”

The company claims that it can produce cell lines in less than a week with 100 percent purity, versus the mixed bags from other companies cell cultures.

“Our moonshot goal is to develop a platform capable of producing every human cell type. This is possible once we understand the genes governing human cell behaviour, which ultimately form the ‘operating system of life’,” Kotter said in a statement. “This will unlock a new generation of cell and tissue therapies for tackling cancer, neurodegenerative disorders and autoimmune diseases and accelerate the development of effective drugs for a range of conditions. The support of leading deep tech and biotech investors will catalyse this unique convergence of biology and engineering.”

[slowlyteenagestarlight]

Don't put this on Facebook unless you want a ban.

Read this!!! 😳

Someone's BRILLIANT HOMEWORK!!!!!

“I made a list of all vaccine ingredients and contacted Poison Control. After intros and such, and asking to speak with someone tenured and knowledgeable, this is the gist of the conversation.

Me: My question to you is how are these ingredients categorized? As benign or poison? (I ran a few ingredients, formaldehyde, Tween 80, mercury, aluminum, phenoxyethanol, potassium phosphate, sodium phosphate, sorbitol, etc.)

He: Well, that's quite a list... but I'd have to say easily that they're all toxic to humans... Used in fertilizers, pesticides... To stop the heart... To preserve a dead body... They're registered with us in different categories, but pretty much poisons. Why?

Me: If I were deliberately to feed or inject my child with these ingredients often, as a schedule, obviously I'd put my daughter in harm's way... But what would legally happen to me?

He: Odd question... but you'd likely be charged with criminal negligence... perhaps with intent to kill... and of course child abuse... Your child would be taken away from you... Do you know of someone's who's doing this to their child? This is criminal...

Me: An industry... these are the ingredients used in vaccines... with binding agents to make sure the body won't flush them out... to keep the antibody levels up indefinitely...

He: WHAT?!

Your conclusion?

The man was beside himself. He asked if I would email him all this information. He wanted to share it with his adult kids who are parents. He was horrified and felt awful he didn't know... his kids are vaccinated and they have health issues...”

~ By Iris Figueroa 🍃♥️🍃

✅INGREDIENTS IN VACCINES - You CANNOT make an educated decision without being educated.

Here are just SOME vaccine ingredients.

These are being INJECTED into your kids;

◾️Formaldehyde/Formalin - Highly toxic systematic poison and carcinogen.

◾️Betapropiolactone - Toxic chemical and carcinogen. May cause death/permanant injury after very short exposure to small quantities. Corrosive chemical.

◾️Hexadecyltrimethylammonium bromide - May cause damage to the liver, cardiovascular system, and central nervous system. May cause reproductive effects and birth defects.

◾️Aluminum hydroxide, aluminum phosphate, and aluminum salts - Neurotoxin. Carries risk for long term brain inflammation/swelling, neurological disorders, autoimmune disease, Alzheimer's, dementia, and autism. It penetrates the brain where it persists indefinitely.

◾️Thimerosal (mercury) - Neurotoxin. Induces cellular damage, reduces oxidation-reduction activity, cellular degeneration, and cell death. Linked to neurological disorders, Alzheimer's, dementia, and autism.

◾️Polysorbate 80 & 20 - Trespasses the Blood-Brain Barrier and carries with it aluminum, thimerosal, and viruses; allowing it to enter the brain.

◾️Glutaraldehyde - Toxic chemical used as a disinfectant for heat sensitive medical equipment.

◾️Fetal Bovine Serum - Harvested from bovine (cow) fetuses taken from pregnant cows before slaughter.

◾️Human Diploid Fibroblast Cells - aborted fetal cells. Foreign DNA has the ability to interact with our own.

◾️African Green Monkey Kidney Cells - Can carry the SV-40 cancer-causing virus that has already tainted about 30 million Americans.

◾️Acetone - Can cause kidney, liver, and nerve damage.

◾️E.Coli - Yes, you read that right.

◾️DNA from porcine (pig) Circovirus type-1

◾️Human embryonic lung cell cultures (from aborted fetuses)

✳️You can view all of these ingredients on the CDCs website. I encourage everyone to do their own research. Look up the MSDS on these chemicals. Read the thousands of peer reviewed studies that have evaluated the biological consequences these chemicals can have on the body, especially when being injected.

✳️Fact check vaccine ingredients here:

https://www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table-2.pdf

PLEASE SHARE!!!

www.cdc.gov

cdc.govDon't put this on Facebook unless you want a ban.

Read this!!! 😳

[biomedgrid]

Biomed Grid | A Review: Targeted Cancer Therapy as a Fight Against Brain Tumor

The Brain tumor can be treated by surgery, radiation, and chemotherapy. With the implications of all these strategies, the overall survival rate of a glioblastoma patient is only 14.6 months after diagnosis [1] . Moreover, the brains of children are very sensitive to tolerate these intense treatment strategies. So, there is a need to discover such agents that particularly target the malignant cells while causing no harm to normal cells [2] . Treatment of brain tumor is very difficult due to certain barriers that make it difficult for the therapeutic agent to reach the targeted infectious cell or tissue.

The major obstacle is the Brain Blood Barrier (BBB) that does not allow any drug or foreign substance to reach to the brain [3]. Another barrier is the enzymes present in BBB e.g. phosphatase enzyme that attack these molecules due to these barriers. So, most drugs have no access to the CNS [4]. Now the particles having to target specificity, i.e. antibody/Nanoparticles can cross this barrier and can reach these affected cells [5]. But the passage of targeted drugs. i.e. protein, oligonucleotides, peptides is interrupted very often [6].

With the identification of tumor-specific antigens, it became possible to develop such therapeutic methods that selectively target diseased cells. Targeted therapies are being used for the treatment of the brain tumor that specifically targets the pathways that are responsible for the disease [7].

Case Report

Brain tumor includes diminishing distinction and deviant multiplication of precursor cells. Recent research has described that cancer-causing events make the change from asymmetrical to symmetrical cell division. The most common malignant tumors in adults and children are Glioma and Medulloblastoma (MB). Glioma is further characterized to Grade I, II, III and grade IV [8]. Grade I is called Pilocytic astrocytoma and mostly occur in children. On the other hand, Grade II and III termed as oligodendroglia astrocytoma, oligoasrocytoma ependymoma is reported to occur in both adults and children’s equally [5] same as vast depict of MBs has accepted numerous subgroups of tumors [9]. Inspire of Glioma and MB cells exhibit definite glial and neuronal makeup, both types of brain tumors have been delineated to emerge from neural indication, competent to show neurogenesis. The ventricle present between the forebrain and spinal canal form glial cells [10]. Glial Fibrillary Acid Protein (GFAP)showing neural stem cells (NSCs) that are also termed as Type B1 cells are considered to sustain asymmetrical cell division to produce transit magnifying ancestors (TAPS, Type C cells), that stimulate differentiation into unripe neuroblast [11].

Obstacles in brain tumor treatment

There are many obstacles to the treatment of brain tumor due to the complex nature of the brain, different morphological characteristics of each type of brain tumor, inaccessibility of therapeutic agents to the tumor cells, difficulty in specifically targeting of tumor cells while giving no harm to surrounding normal cells and sometimes due to the development of resistance against chemotherapeutics agents [12]. One of the major obstacles is the brain blood barrier.

Brain blood barrier

Brain blood barrier (BBB) which protects the brain can be considered as the major obstacle to the therapeutic agent [3]. It includes endothelial cells, pericytes, astrocytes, and neurons. Brain capillary endothelial cells (BCEC) are the major boundary between the brain and cellular tissues. The function of pericytes is to maintain and support the BCEC [13]. Astrocytes are the nonneuronal cells that release signal to regulate the permeability of endothelium [1] . Neurons supply capillary to the BBB with nerves. So, the major barrier component is BCEC [14]. Another barrier is the enzymes present in the brain blood e.g. phosphatase enzyme that attack foreign molecules and limit their access to the CNS [6]. With the formation of brain tumor, infectious cells begin to attack healthy brain tissue and after growing to a certain volume they also give harm to the BBB and formation of blood-brain tumor barrier occurs [15]. Another barrier that limits the entry of therapeutic agents is the blood-cerebrospinal fluid barriers (CSF) [16]. It is also composed of epithelial cells which form a barrier and do not allow any therapeutic agent to enter the brain parenchyma. So, most of the molecules failed to reach the targeted site [17].

Targeted strategies for the Treatment of Brain Tumor

The mediate general survival of patients with glioblastoma (GBM) is only 14.6 months after recent multimodal treatment aggressive surgical abscission followed by coincident or consecutive radiation temozolomide chemotherapy [18].

BBB targeting strategies

The Blood Brain Barrier is not only responsible for the protection of the brain from harmful substances but is also involved in providing the necessary nutrients to the brain. It is the main challenge in drug delivery to the brain [15].

Absorptive-Mediated transcytosis

It furnishes a way for the transport of drugs beyond BBB by cationic protein or cell-Penetrating peptide. Classic cationic bovine serum albumin accumulated with pegylated nanoparticle (CBSANP) was designed for brain targeting drug delivery [19]. It was stated that plasmid PORF-hTRAIL (pDNA) subsume (CBSA-NP) abreast with glycoprotein in brain and tumor micro variability and assembled in tumor cells [19,20] arrogate other cationic protein called wheat germ agglutinin (WGA) associated to the surface of liposomes and also reveal elevated BBB transporter.CPP has been used to lessen the lipophilic barrier of the cell membrane and transport large diversity of baggage which involves peptides, DNA, antibodies, toxins and Nano drug carrier such as liposomes and micelles [21].

Transporter-mediated transcytosis

It is substrate selective so the drugs that intimate the endogenous substances would be delivered to the brain. Glucose Transporters (GLUT) which ease the delivery of glucose from the blood to the brain, have inclusive approach use in brain quarry. Another chief system is choline transporter which ties up positively charged ammonium group or simple cations [22]. The glazed nanoparticle was skill full to hybrid an in vitro replica of BBB (Bovine BCEC).

Receptor-mediated transcytosis

This is the technique that involves the only drug delivery that bosom imitates the endogenous carrier and then delivered to the brain. One of the most accepted receptor-mediated transcytoses for brain targeting is the Transferrin Receptor (TfR) [23]. These are stacked with polyphosphoester Hybrid Micelles (TPM). It expresses a strong anti- Glioma activity [11]. The low-density lipoprotein (LDC) and Receptor-related protein (LRP) have been noticed to mediate delivery of different ligand associated with Nano carrier beyond the BBB [24].

BBTB targeting strategies and drug delivery

It is present between brain tumor tissues and micro vessels devised by extremely peculiar endothelium cells (ECs) restricting the transforming of the hydrophilic molecule to tumors [25]. The techniques suggested for BBTB are mostly constructed on receptor highly intimate on the tumor as epidermal growth factor receptor and integrity. Arginine glycine aspartic acid (RGD) is going to be used for targeted drug transformation to the brain.

Antibody-Targeted Therapeutics

 Antibodies can be used for this purpose, which can easily identify the foreign toxic substance. It can easily cross multiple barriers including BBB and selectively accumulate in tumor cells [26]. As in the brain tumor, new blood vessels are also formed termed as angiogenesis assisting in growth and proliferation of brain tumor. These molecular targets inhibit angiogenesis as a treatment of the brain tumor. An antibody named as Bevacizumab VEGF (Vascular endothelial growth factor) is very efficient because it targets the mediator of angiogenesis [1] . Cetuximab IgG1 antibody can block this receptor [1] . EPGR (Epidermal growth factor receptor) show interaction with the protein kinase C which signals to activate transcription. Panitimumab and Erlotinib can also be used as an inhibitor of EPGR. Erlotinib can be used in combination with radiotherapy to treat glioma and be tolerated by the patients. It can be taken orally. Nimotuzumba can also especially target tumor cells due to the overexpression of EPGR and give no harm to cancer cells [2] .

Nanoparticle targeted therapy

Nano Technology put great work on brain Tumor and Open the doors for future treatments and researches for the brain tumor. Nano Particles especially play a distinct role in Brain tumor’s treatment, surgery and pharmacology researches. Today researches in Nanoparticles have proved a major contributor role for more future work on the brain tumor.

Importance of Nano Particles

Nanoparticles are important in the treatment of the brain tumor because they can be used by two mechanisms:

a. Specific mechanisms

b. Nonspecific mechanisms

Specific mechanism based on the interaction of antigens on the surface of nanoparticles with tumor cell receptors. Non-specific mechanism based on preference extravasation of nanoparticles that access the blood-brain barrier (BBB or tumor). Nanoparticles only access the tumor because they have more retention effects than other particles that do not access the tumor.

Peptide vaccine

In nanoparticle, peptide vaccine helps to target the single or multiple Nano antigens. Some peptide vaccines are EGFRviii, IL- 13RA2, AIM-2, and MAGE-19 [27,28]. These vaccines are more responsive in patients and tumor patients survived 26 months on average. Peptide vaccine can cause profusion of the cell which does not indicate the targeted antigens. Targeting multiple antigens was the most effective treatment of the tumor [29].

Administration Techniques for Molecular Therapeutics

Molecular targeting agents can be administrated toward the targeted site by systematic, intracranial and cell-mediated delivery.

      Systematic administration

Systematic Administration of targeted therapeutics is a very easy technique for cargo transportation involving dreary quantity [13]. One evident technique for transportation of these molecules is through Intravenous (IV) injections. The nanoparticle established remedial are frequently administrated in many doses to control the growth of the tumors [30]. Low-frequency enrapt ultrasound supplies confined interference of BBB [31-33]. The preclinical studies have shown that this technique can purely increase the central delivery of the remedial agents into tumor cells [31]. Another method to ease the non -invasion technique is Magnetic targeting at a specialized site [34,35] has been practiced in clinical trials (Clinical Trails Gov. Identifier NCT0005495, NCT00034333). It is reported that magnetic targeting produces a 5times enhancement in the total subjection of glioma cells to the nanoparticle over not specific and non- targeting tumors and increase in target selection for collection in the tumor versus normal brain cells [13]. The main drawbacks of systemic delivery are the danger for the accumulation of nanoparticle in Nano-targeted organs like the liver, kidney, and lungs. The nanoparticle is not hazardous but the longtime interaction of nanoparticle deposition in the brain has not fully addressed [36].

Intracranial administration

Delivery of therapeutic agent to the targeted site in the brain tumor is a hard job without affecting normal cells. We can enter a therapeutic agent directly into the tumor site that can easily pass the BBB barrier [37]. We can use polymers that are capable to target tumor cells while sparing normal cells. These polymers can be degradable or non-degradable [38]. This method is mostly used when localization of drug to the target is low. It increases the concentration of such an agent within affected cells only. It depends on transporting a large amount of therapeutic agent in a continuous flow [39].

Cell-mediated administration

Cell-mediated delivery of synthetic nanoparticle-based drug carrier is a very promising strategy to control the distribution of the drug and improve targeting. In this method, cells are found the ability to cross the tumor. Mesenchymal stem cell (MSC) and neural stem cells show the ability to target the brain tumor cells by nanoparticle.

MSCs with bound silica Nano rattle-doxorubicin can be targeted in vivo and in vitro ways on the tumor [40]. It was noticed that Nano capsule containing ferrociphenol can be entered into the tumor and can slow the growth of the tumor [41].

The ability of ph-mediated drug release capabilities. This can deliver doxorubicin conjugated to BBB. This method of combining stem cells and nanoparticle for treatment based upon local administration method.[36]

Another way of treatment is that to combine the carriers and nanoparticles and carriers are tracked via trucker. For this purpose iron oxide was used [40].

Issues and Limitations of Brain-Targeted Therapies

Development of targeted vehicle is only possible after complete understanding of the target. As the brain is complex in nature so it is very difficult to recognize the specific differences between the target and other cells [6]. Non-covalent drug delivery system often results in the release of drug at a non-specific point during its transport. Some targeted molecules are also degraded by the enzymes. Another problem is safety. Some of the targeted molecules cause toxicity problems. e.g. Nanoparticles show slow toxicity after their entry in the systemic circulation and their mechanism of action are not fully known. So, there is no idea of the appearance of any side effect due to their long-term accumulation on the targeted site. Most of the targeted approaches are used with conventional methods of treatment i.e. chemotherapy and radiotherapy for disease management [13]. It limits the use of targeted molecules that target a number of pathways involved in the formation of tumor i.e. broad ranging therapeutic agent must be used [2] .

Conclusion

A number of brain tumor patients are being treated by the targeted therapies which have many advantages over conventional treatment techniques e.g. surgery, radiation, and chemotherapy. The therapeutic agents have many advantages and one of the potential advantages is that these can easily cross the barriers that protect the brain and target only the malignant site and is not harmful to the normal healthy cells of the brain. But it also has some dark aspects i.e. sometimes, it causes non-specific transport, and also sometimes before reaching the site of action these are degraded by the enzymes, and, toxicity profiles. Although these therapeutic agents have some issues and limitations they are mostly used in combination with conventional techniques to improve their efficacy.

Read More About this Article: https://biomedgrid.com/fulltext/volume3/a-review-targeted-cancer-therapy-as-a-fight-against-brain-tumor.000672.php

For more about: Journals on Biomedical Science :Biomed Grid