The document describes a research study aimed at developing biomarkers for detecting potential allergenicity of novel foods, including genetically modified foods. The researcher conducted experiments challenging mice with known food allergens (egg ovomucoid protein and peanut protein) and analyzed gene expression profiles in the mice spleens. Several hundred genes were found to be differentially expressed. After validating some genes, the researcher identified potential biomarker genes that could help detect allergenicity of GM foods. The study provides insights into transcriptomic responses to food allergens and biomarkers that may help evaluate allergenicity of novel foods like GM crops.
Cow´s milk allergy is a common food allergy especially among infants and young children. The major allergen in cow´s milk is beta-lactoglobulin (BLG) and BLG-specific antibodies are needed for diagnostic, therapeutic, food processing and quality verification applications. At VTT we have established a IgE antibody library from a milk allergic person and identified high-affinity antibodies against native and heat denaturated BLG. As an alternative production system, we have used barley as a production host. Proof-of-concept was gained with expressing the BLG-specific antibody in barley cell culture. Best production levels in barley grains were obtained with glutelin-specific promoter combined to ER targeting and retention signals. The barley-produced BLG-specific antibody was successfully purifed with affinity-based chromatography and the functionality of barley-produced BLG-specific antibody was verified by ELISA and immunoprecipitation assays. The developed barley-based expression system clearly demonstrated its potential for application in the processing of dairy milk products as well as in detecting allergens from foods possibly contaminated by cow´s milk.
Next generation biotherapeutics production system Trichoderma reeseiChristopher Landowski
The filamentous fungus Trichoderma reesei is an important production organism used by industrial enzyme companies world-wide. It is a low cost production system that secretes its native enzymes at levels exceeding 100 g/L of culture medium. Several T. reesei produced enzymes have obtained the generally recognized as safe status by the U.S. Food and Drug Administration. T. reesei has tremendous prospects to be a cost efficient and high yield system for producing therapeutic proteins. We have adapted the fungus to become more suitable for biotherapeutic production by reducing secreted protease activity and altering glycosylation pathways needed for adding mammalian glycoforms.
Expression strains for monoclonal antibodies, Fab antibody fragments, interferon alpha-2b, insulin-like growth factor 1, and fibroblast growth factor 21 were constructed, cultivated in bioreactors, and expression levels were measured from the culture medium. After deleting 13 of the most critical protease genes, the general secreted protease activity was reduced over 30-fold. Monoclonal antibodies could be produced up to 7.6 g/L, Fab antibody fragments up to 8.2 g/L, interferon alpha-2b at 7.9 g/L, and insulin-like growth factor fusion protein at 8 g/L. With protease inhibitor treatment interferon alpha-2b could be produced at over 10 g/L, insulin-like growth factor fusion protein at 19 g/L, and full length fibroblast growth factor 21 at 200 mg/L in addition to a shorter form at 3.5 g/L. Human glycoforms such as G0 and FG0 were produced on monoclonal antibodies.
Expression levels and product quality improved dramatically after multiple protease deletions and optimization of culture conditions. While the production levels achieved are already relatively high, the strains could be developed further to reach the 100 g/L potential of the organism. This study demonstrates the excellent prospects of T. reesei as a host for therapeutic protein production.
Cow´s milk allergy is a common food allergy especially among infants and young children. The major allergen in cow´s milk is beta-lactoglobulin (BLG) and BLG-specific antibodies are needed for diagnostic, therapeutic, food processing and quality verification applications. At VTT we have established a IgE antibody library from a milk allergic person and identified high-affinity antibodies against native and heat denaturated BLG. As an alternative production system, we have used barley as a production host. Proof-of-concept was gained with expressing the BLG-specific antibody in barley cell culture. Best production levels in barley grains were obtained with glutelin-specific promoter combined to ER targeting and retention signals. The barley-produced BLG-specific antibody was successfully purifed with affinity-based chromatography and the functionality of barley-produced BLG-specific antibody was verified by ELISA and immunoprecipitation assays. The developed barley-based expression system clearly demonstrated its potential for application in the processing of dairy milk products as well as in detecting allergens from foods possibly contaminated by cow´s milk.
Next generation biotherapeutics production system Trichoderma reeseiChristopher Landowski
The filamentous fungus Trichoderma reesei is an important production organism used by industrial enzyme companies world-wide. It is a low cost production system that secretes its native enzymes at levels exceeding 100 g/L of culture medium. Several T. reesei produced enzymes have obtained the generally recognized as safe status by the U.S. Food and Drug Administration. T. reesei has tremendous prospects to be a cost efficient and high yield system for producing therapeutic proteins. We have adapted the fungus to become more suitable for biotherapeutic production by reducing secreted protease activity and altering glycosylation pathways needed for adding mammalian glycoforms.
Expression strains for monoclonal antibodies, Fab antibody fragments, interferon alpha-2b, insulin-like growth factor 1, and fibroblast growth factor 21 were constructed, cultivated in bioreactors, and expression levels were measured from the culture medium. After deleting 13 of the most critical protease genes, the general secreted protease activity was reduced over 30-fold. Monoclonal antibodies could be produced up to 7.6 g/L, Fab antibody fragments up to 8.2 g/L, interferon alpha-2b at 7.9 g/L, and insulin-like growth factor fusion protein at 8 g/L. With protease inhibitor treatment interferon alpha-2b could be produced at over 10 g/L, insulin-like growth factor fusion protein at 19 g/L, and full length fibroblast growth factor 21 at 200 mg/L in addition to a shorter form at 3.5 g/L. Human glycoforms such as G0 and FG0 were produced on monoclonal antibodies.
Expression levels and product quality improved dramatically after multiple protease deletions and optimization of culture conditions. While the production levels achieved are already relatively high, the strains could be developed further to reach the 100 g/L potential of the organism. This study demonstrates the excellent prospects of T. reesei as a host for therapeutic protein production.
Antioxidant-mediated up-regulation of OGG1 via NRF2 induction is associated ...Enrique Moreno Gonzalez
Estrogen metabolism-mediated oxidative stress is suggested to play an important role in estrogen-induced breast carcinogenesis. We have earlier demonstrated that antioxidants,
vitamin C (Vit C) and butylated hydroxyanisole (BHA) inhibit 17β-estradiol (E2)-mediated oxidative stress and oxidative DNA damage, and breast carcinogenesis in female August
Copenhagen Irish (ACI) rats. The objective of the present study was to characterize the mechanism by which above antioxidants prevent DNA damage during breast carcinogenesis.
Answer four fundamental questions on how to develop the most innovative cancer immunotherapy treatments, starting with screening for lead molecules and ending with evaluation of combination therapies.
Food as Medicine:solution for chronic inflammationKevin KF Ng
Food as medicine, chronic inflammation, life expectancy, causes of death, unifying theory of degenerative diseases and cancer, c-reactive protein, risks factors associated with elevated CRP, interventions to lower CRP, benefits of lowering CRP
Antioxidant-mediated up-regulation of OGG1 via NRF2 induction is associated ...Enrique Moreno Gonzalez
Estrogen metabolism-mediated oxidative stress is suggested to play an important role in estrogen-induced breast carcinogenesis. We have earlier demonstrated that antioxidants,
vitamin C (Vit C) and butylated hydroxyanisole (BHA) inhibit 17β-estradiol (E2)-mediated oxidative stress and oxidative DNA damage, and breast carcinogenesis in female August
Copenhagen Irish (ACI) rats. The objective of the present study was to characterize the mechanism by which above antioxidants prevent DNA damage during breast carcinogenesis.
Answer four fundamental questions on how to develop the most innovative cancer immunotherapy treatments, starting with screening for lead molecules and ending with evaluation of combination therapies.
Food as Medicine:solution for chronic inflammationKevin KF Ng
Food as medicine, chronic inflammation, life expectancy, causes of death, unifying theory of degenerative diseases and cancer, c-reactive protein, risks factors associated with elevated CRP, interventions to lower CRP, benefits of lowering CRP
The Documented Health Risks of Genetically Engineered Foods (Long Version)Jack Olmsted
Content of the Presentation
Regulatory Failure: It is useful to explain how such dangerous products could have made it to the market with government approval. Several slides include quotes from formerly secret FDA documents that show how government policy was at odds with more cautious scientific opinion at the agency.
Health Risks of GMOs: This section highlights many of the adverse findings revealed through laboratory experiments and reported by farmers, doctors, and investigators. It also introduces some theoretical risks based on the current state of the science.
The Consumer Tipping Point: The final section includes a discussion of a strategy to achieve the tipping point of consumer rejection of GMOs in the US, which is the basis for our Campaign for Healthier Eating in America. The key elements needed are consumer education on GMO health risks combined with clear non-GMO choices.
The Institute for Responsible Technology http://www.responsibletechnology.org/resources/powerpoint-presentation-on-gmos
Presentation on biotech in agriculture for educational purposes. Thanks to Kevin Folta. I borrowed his concept of showing who benefits from various traits from one of his excellent slide sets.
Why take NUTRILITE® DOUBLE X® Vitamin/Mineral/Phytonutrient? Because experts recommend
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Do you have that unnecessary middle fats ? This belly fat is sometimes known as abdominal obesity. Sitting jobs, lack of exercise and unhealthy food lead to increase this middle fat. Check out here simple and effective ways to treat abdominal obesity.
Cloning and expression of Human glutamic acid decarboxylase (GAD 65) gene in ...Open Access Research Paper
Diabetes is a chronic autoimmune disease characterized by the inability of body to produce or respond to insulin a hormone required by body to burn glucose for energy. Type I Diabetes mellitus, also known as Insulin Dependent Diabetes mellitus is a most frequent chronic disease of childhood, afflicts 0.2-0.3% of human individuals due to auto immune destruction of insulin secreting pancreatic β cells. GAD65 is the major auto antigen in Insulin Dependent Diabetes Mellitus (IIDM). Thus, this project is aimed at expression of GAD65 in E. coli. GAD65 gene was cloned into pET-28a bacterial expression vector and expression was studied in BL21 DE3 cells. Different parameters of induction like isopropyl-β-D-thiogalactopyranoside (IPTG), temperature, time interval were standardized. The recombinant clones induced with 2 μM of IPTG at 30oC for 4 h at flask level produced the protein upto 537μg/ml. Furthermore, the specificity of the purified recombinant protein was confirmed by western blot analysis using monoclonal antibodies. This work establishes a strategy in E. coli for the expression of GAD65 with optimized parameters.
The Lyme disease agent, borrelia burgdorferi, colonizing the gut of the tick lxodes scapularis,can transmit pathogens to vertebrate hosts including humans. B. burgdorferi colonization increases the expression of several tick gut genes including pixr. Abrogation of PIXR function in vivo alters the gut microbiome, metabolome and immune responses. Changes in the gut microbial members are likely to influence the metabolome of the tick gut due to differences in the metabolic functions unique to the specific bacteria genera. Changes in the composition of intestinal metabolites can be analyzed by means of untargeted metabolomics. https://www.creative-proteomics.com/services/untargeted-metabolomics.htm
Application of proteomics for identification of abiotic stress tolerance in c...Vivek Zinzala
It is the study of “Proteome”.
The word "proteome" is a blend of "protein" and "genome”.
Large scale study of Proteins.
Particularly their structures and functions.
Study of full set of proteins in a cell type or tissue, and changes during various conditions
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Detection and Surveillance of Antibiotic Resistance Genes From Food and Ferti...QIAGEN
One potential way to acquire antibiotic resistance genes is through the food supply chain. Both livestock and feed may
acquire antibiotic resistant bacteria via different mechanisms. Foodstuffs can be exposed to antibiotic resistant bacteria
through fertilizer originating from waste-water treatment plants. This, in addition to increasing administration of antibiotics
to livestock, can lead to food being a potential source of antibiotic resistance genes. This may lead to horizontal gene
transfer to pathogenic enteropathogens and further to drug resistance in humans. Therefore, the surveillance and prevention
of antibiotic resistance genes in food is important.
To effectively combat the spread of difficult-to-treat bacterial infections, rapid surveillance methods to detect antibiotic
resistance genes are required; in order to monitor both bacterial isolates and metagenomic samples.
Since the gut is known to act as a reservoir for antibiotic resistance genes, a small-scale research study was performed on
5 stool samples isolated from healthy human adults using an antibiotic resistance gene identification PCR array. In addition,
the diversity of antibiotic resistance genes in municipal biosolids was determined using an Antibiotic Resistance Genes
Microbial DNA qPCR Array with DNA extracted from belt-filter, press-cake sewage samples.
22 antibiotic resistance genes were identified from different resistance classifications. Further studies were performed in
beef, chicken, vegetable and pork samples. In conclusion, PCR arrays can be effective tools for detection of antibiotic
resistance genes from food samples and potential fertilizer sources.
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
Andreas Schleicher presents at the OECD webinar ‘Digital devices in schools: detrimental distraction or secret to success?’ on 27 May 2024. The presentation was based on findings from PISA 2022 results and the webinar helped launch the PISA in Focus ‘Managing screen time: How to protect and equip students against distraction’ https://www.oecd-ilibrary.org/education/managing-screen-time_7c225af4-en and the OECD Education Policy Perspective ‘Students, digital devices and success’ can be found here - https://oe.cd/il/5yV
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptx
GM Food Allergy Biomarkers
1. Development of Biomarkers for the Detection of Potential Allergenicity of Novel Food Mainul Husain Department of Animal & Poultry Science University of Guelph Ontario, Canada
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7. Overall Project Design Validation of Candidate Bio-Marker Genes By Real Time RT- PCR Data Analysis Mapping Genes To Functional Pathways Allergen Treated Experimental Mice Control Mice Total RNA Extracted From Spleens Total RNA Extracted From Spleens Pooled Total RNA Pooled Total RNA SELECTION OF GENES AS ALLERGY BIO-MARKERS Affymetrix Microarray Classification of Genes Using Gene Ontology
8. Balb/c Mice Challenged With Egg Ovomucoid Protein Total 30 Female BALB/c Mice In Control and Treated Groups 15 Control Mice Challenged with Amino Acid Solution (1mg/100 l+10 g CT) 15 Experimental Mice Challenged with Ovomucoid (1mg/100 l+10 g CT) Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Pooled Total RNA Pooled Total RNA Pooled Total RNA Pooled Total RNA Pooled Total RNA Pooled Total RNA
13. KEGG Pathways Affected In Response to Ovomucoid Challenge Pathways Number of Genes Identified MAPK SIGNALING PATHWAY 27 OXIDATIVE PHOSPHORYLATION 24 FOCAL ADHESION 24 CYTOKINE-CYTOKINE RECEPTOR INTERACTION 24 REGULATION OF ACTIN CYTOSKELETON 20 WNT SIGNALING PATHWAY 19 CELL CYCLE 18 PURINE METABOLISM 16 INSULIN SIGNALING PATHWAY 15 PENTOSE AND GLUCURONATE INTERCONVERSIONS 13 PORPHYRIN AND CHLOROPHYLL METABOLISM 13 STARCH AND SUCROSE METABOLISM 13 METABOLISM OF XENOBIOTICS BY CYTOCHROME P450 13 T CELL RECEPTOR SIGNALING PATHWAY 13 JAK-STAT SIGNALING PATHWAY 13 HEMATOPOIETIC CELL LINEAGE 12 AXON GUIDANCE 12 ANDROGEN AND ESTROGEN METABOLISM 11 TGF-BETA SIGNALING PATHWAY 11 RIBOSOME 11 ADHERENS JUNCTION 10 PHOSPHATIDYLINOSITOL SIGNALING SYSTEM 10
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15. 15 Control Mice Treated with Amino Acid Solution (1mg/100 l + 10 g CT) C3H/HeJ Mice Challenged With Peanut Protein Total 30 C3H/HeJ Mice In Both Control and Treated Groups 15 Experimental Mice Challenged with Peanut Protein (1mg/100 l + 10 g CT) Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA RNA Pool (5 random samples) RNA Pool (5 random samples) RNA Pool (5 random samples) RNA Pool (4 High Score Samples) RNA Pool (4 Low Score Samples) RNA Pool (4 High Score Samples) RNA Pool (3 Low Score Samples)
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17. Biological Classification of The Significant Genes Ontology ID Gene Ontology Terms # of Genes P-Value FDR 44238 Primary Metabolism 467 <0.0001 <0.0001 44237 Cellular Metabolism 475 <0.0001 <0.0001 43283 Biopolymer Metabolism 204 <0.0001 <0.0001 8152 Metabolism 504 <0.0001 <0.0001 7582 Physiological Process 705 0.0001 0.0023 43170 Macromolecule Metabolism 296 <0.0001 0.0025 50875 Cellular Physiological Process 650 0.0001 0.0029 6323 DNA Packaging 29 0.0002 0.0069 6325 Establishment and/or Maintenance of Chromatin Architecture 28 0.0003 0.0076 6259 DNA Metabolism 54 0.0006 0.0395 6139 Nucleobase, Nucleoside, Nucleotide And Nucleic Acid Metabolism 227 0.0007 0.0405 7001 Chromosome Organization and Biogenesis 29 0.0014 0.0700 16568 Chromatin Modification 20 0.0012 0.0705 6461 Protein Complex Assembly 18 0.0022 0.1158 8652 Amino Acid Biosynthesis 8 0.0037 0.1167 51244 Regulation of Cellular Physiological Process 215 0.0035 0.1176 51128 Regulation of Cell Organization and Biogenesis 10 0.0026 0.1186 6470 Protein Amino Acid Dephosphorylation 17 0.0025 0.1193 15031 Protein Transport 60 0.0027 0.1193 19538 Protein Metabolism 211 0.0034 0.1194
18. Directed Acyclic Graph (DAG) of Enriched Immune Response Category Some other significantly enriched sub-categories are: (1) Innate immune response, (2) Positive regulation of inflammatory response, (3) T-cell proliferation, (4) B-cell proliferation, (5) Positive regulation of interferon-beta biosynthesis.