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Technical Notes

Journal Club #11: Elovanoids as a Potential Anti-Allergen in an In Vitro HDM Model

Journal Club #11: Elovanoids as a Potential Anti-Allergen in an In Vitro HDM Model

HNEpC were challenged with different stressors or HDM extract for 30 minutes, after which they were then treated with elovanoids. Cytotoxicity, cell viability, and the expression of pro-inflammatory cytokines and chemokines in the cell supernatant were evaluated. The study concluded that the HDM challenge in the cellular experimental allergy model induced expression of several key cytokines and chemokines linked to respiratory diseases and the treatment with elovanoids reduced cell toxicity and pro-inflammatory cytokine levels while increasing cell viability and anti-inflammatory cytokine levels. Overall, elovanoids might constitute a therapeutic option in the prevention and treatment of allergies to HDM. Read More

Journal Club #10: The Roles of High Mobility Group Box 1 (HMGB1) and Toll-like Receptor 4 (TLR4) in Arthritis-Induced Pain in the Collagen-Antibody Induced Arthritis Model

Journal Club #10: The Roles of High Mobility Group Box 1 (HMGB1) and Toll-like Receptor 4 (TLR4) in Arthritis-Induced Pain in the Collagen-Antibody Induced Arthritis Model

The collagen antibody-induced arthritis (CAIA) model was used for investigating if blockade of peripheral HMGB1 attenuates pain-like behavior in a sex-dependent fashion. Both male and female CAIA mice revealed high HMGB1 mRNA levels in the ankle joints. Administering HMGB1 neutralizing antibodies failed to reduce arthritis severity in both male and female mice; however, it did significantly reverse CAIA-induced mechanical hypersensitivity in male mice. In addition, the differences in responses between sexes were evaluated by measuring cytokine levels (TNF, IL-6, CXCL1 and CXCL2) from cultured macrophages treated with HMGB1 and cytokine mRNA levels (TNF, IL-1b, IL-6, CCL2, CXCL1, and CXCL2) in paws which received HMGB1. Read More

Journal Club #9: Evaluating Decellularized ECM Hydrogels to Support Brain Organoid Culture

Journal Club #9: Evaluating Decellularized ECM Hydrogels to Support Brain Organoid Culture

The effectiveness of decellularization (DC) process in removing cells from the ECM by chemical, physical, and enzymatical methods were investigated using several assays. DC protocols were evaluated for their effectiveness in removing blood and other soluble materials and optimized after comparing combinations of DNA-removing enzymes with different detergents such as SDS, Triton X, and sodium deoxycholate (SDC). SDC proved to be the most suitable DC detergent after evaluating the quantified collagen and GAG levels in the decellularized ECMs. Read More

Journal Club #8: The Role of Epitope Retrieval in the Production of Anti-Glomerular Basement Membrane Antibodies in the Antibody-Associated Vasculitis Model

Journal Club #8: The Role of Epitope Retrieval in the Production of Anti-Glomerular Basement Membrane Antibodies in the Antibody-Associated Vasculitis Model

The anti-alpha3(IV)NC1 mAb failed to bind to the GBM and the basement membrane of Bowman’s capsules in formalin-fixed, paraffin-embedded normal kidney sections without antigen retrieval using immunohistochemistry. However, the mAb bound to retrieved antigens which received heat treatment under acidic conditions, a treatment of proteases from S. griseus or neutrophil elastase, but not with heat treatment under neutral conditions and alkaline conditions. This suggests that ANCA activated neutrophils may digest type IV collagen and expose α3(IV)NC1 as an antigen which then induces production of anti-GBM antibodies. Read More

Journal Club #7: Characterizing the Gastrointestinal (GI) Abnormalities in the Mouse Dystonia Musculorum (DSTdt) Model for Hereditary Sensory and Autonomic Neuropathy Type VI (HSAN-VI)

Journal Club #7: Characterizing the Gastrointestinal (GI) Abnormalities in the Mouse Dystonia Musculorum (DSTdt) Model for Hereditary Sensory and Autonomic Neuropathy Type VI (HSAN-VI)

Mutation of dystonin gene in mice induces hereditary sensory and autonomic neuropathy type VI (HSAN-VI) which leads to abnormalities in the gastrointestinal tract and enteric nervous system. These can result in an imbalance of the autonomic control over the gut, but do not affect intracellular transportation of D-Xylose. The mouse model of HSAN-VI can elucidate and further characterize the human disease and determine potential treatments for symptom relief. Read More

Journal Club #6: DPP-4 Inhibition can Improve the Resolution of Glomerular Injury in a Rat Glomerular Nephritis Model

Journal Club #6: DPP-4 Inhibition can Improve the Resolution of Glomerular Injury in a Rat Glomerular Nephritis Model

DPP-4 inhibitor preventive treatments in nephritic rats significantly reduced the number of crescents, glomerulosclerosis, tubular injury, and renal fibrosis compared with that of untreated nephritic rats. In the short-term group (7-14 days), the DPP‐4 inhibition significantly reduced proteinuria and serum urea levels by Day 14 (therapeutic). On the other hand, in the long‐term group (3-8 weeks), the DPP‐4 inhibition resulted in 25% lower proteinuria levels by the 3-5 weeks timepoint (preventative). Furthermore, DPP-4 function and glomeruli fibrosis histology were evaluated by measuring serum chemokine levels and Sirius Red staining, respectively. Read More

Journal Club #5: Osteochondral Differentiation of Mesenchymal Stem Cells in Biodegradable Zinc Oxide Scaffolds as Confirmed by Analyzing Type I and Type II Collagen Production

Journal Club #5: Osteochondral Differentiation of Mesenchymal Stem Cells in Biodegradable Zinc Oxide Scaffolds as Confirmed by Analyzing Type I and Type II Collagen Production

Mesenchymal stem cells cultured on 2.5% ZnO composite scaffold produced the highest type II collagen indicating a mature marker of hyaline cartilage, and low type I collagen associating with immature chondrocytes. In addition to collagen analysis, the ECM were analyzed with cell numbers, glycosaminoglycan levels, as well as several gene expression. Read More

Journal Club #4: Evaluating the Neutralizing Immune Response in SARS-CoV-2 Infection By Serological Assays

Journal Club #4: Evaluating the Neutralizing Immune Response in SARS-CoV-2 Infection By Serological Assays

Antibody responses to the SARS-CoV-2 spike protein receptor binding domain (RBD) in ELISA and Plaque reduction neutralization tests (PRNT) were analyzed in the serum of 150 patients. The results show viral infection elicits robust neutralizing antibody titers in most individuals which last beyond 6 months. Read More

Journal Club #3: Interleukin-4 Receptor alpha Subunit Deficiency Alleviates Intestinal Inflammation in a Mouse Dextran Sulfate Sodium (DSS)-Induced Colitis Model by Enhancing the Intestinal Mucosal Barrier

Journal Club #3: Interleukin-4 Receptor alpha Subunit Deficiency Alleviates Intestinal Inflammation in a Mouse Dextran Sulfate Sodium (DSS)-Induced Colitis Model by Enhancing the Intestinal Mucosal Barrier

Interleukin-4 receptor α subunit knockout mice showed alleviated intestinal inflammation in a dextran sulfate sodium (DSS)-induced colitis model compared to wild-type mice. The remission of disease activities was caused by an improved intestinal mucosal barrier function as evaluated with FITC-Dextran (4kDa). Read More

Journal Club #2: Exosomes From Human Mesenchymal Stem Cells Can Expedite ECM Development as Confirmed by Analyzing GAG and Collagen Production levels

Journal Club #2: Exosomes From Human Mesenchymal Stem Cells Can Expedite ECM Development as Confirmed by Analyzing GAG and Collagen Production levels

Extracellular vesicles from human mesenchymal stem cells which express CD9 and CD63 can expedite chondrogenesis in early-stage 3D human degenerative disc cell cultures. This was confirmed by increased levels of glycosaminoglycan and total collagen production and cell maturation markers such as IL-6 and IL-8 in the culture media. Read More

Journal Club #1: Evaluating Immune Responses in a Mouse House Dust Mite (HDM)-induced Asthma Model Using Mouse Anti-HDM Antibody Assays.

Journal Club #1: Evaluating Immune Responses in a Mouse House Dust Mite (HDM)-induced Asthma Model Using Mouse Anti-HDM Antibody Assays.

Prophylactic effects of five different Lactobacillus casei strains on house dust mite (HDM)-induced asthma were individually evaluated by assaying for anti-HDM IgG1 and IgE levels responsible for the Th2 reaction, and IgG2b levels responsible for the Th1 reaction using anti-HDM antibody assay kits. Read More

Cytokine & Chemokines in Disease: Allergies

Cytokine & Chemokines in Disease: Allergies

Cytokines and chemokines are key modulators of immune responses and play diverse roles in inflammatory diseases. Here, we discuss the role of specific cytokines and chemokines in allergies and hypersensitivity reactions. Read More

Cytokines & Chemokines in Disease: Cancer

Cytokines & Chemokines in Disease: Cancer

Cytokines and chemokines are key modulators of immune responses and play diverse roles in inflammatory diseases. Here, we discuss the role of specific cytokines and chemokines in cancer and tumor metastasis. Read More

Cytokines & Chemokines in Disease: Inflammatory Bowel Disease & Nephritis

Cytokines & Chemokines in Disease: Inflammatory Bowel Disease & Nephritis

Cytokines and chemokines are key modulators of immune responses and play diverse roles in inflammatory diseases. Here, we discuss the role of specific cytokines and chemokines in inflammatory bowel disease (IBD) and membranous nephropathy/nephritis. Read More

Cytokines & Chemokines in Disease: Rheumatoid Arthritis

Cytokines & Chemokines in Disease: Rheumatoid Arthritis

Cytokines and chemokines are key modulators of immune responses and play diverse roles in inflammatory diseases. Here, we discuss the role of specific cytokines and chemokines in rheumatoid arthritis (RA). Read More

Role of Matrix Metalloproteinases in Disease Development

Role of Matrix Metalloproteinases in Disease Development

Matrix metalloproteinases (MMPs) are primarily responsible for the physiological and pathological turnover of tissues throughout the body. While normally expressed in low levels in tissues of healthy adults, MMPs are up-regulated at sites of tissues damage to facilitate tissue repair. Additionally, several disease states, such as osteoarthritis, rheumatoid arthritis, and cancer, also have increased MMP activity. This blog discusses the various MMPs in the progression of those diseases. Read More

Matrix Metalloproteinases (MMPs): Multi-Faceted Family of Endopeptidases

Matrix Metalloproteinases (MMPs): Multi-Faceted Family of Endopeptidases

MMPs are a family of endopeptidases with a wide array of substrates, ranging from collagen and other ECM constituents to cytokines/chemokines, and even other MMPs. As such, MMPs have a complex biological role involving several different signaling pathways. Read here to learn more about basic MMP structure and the role of MMPs in physiological processes. Read More

Andrographolide: Potential Treatment for Ulcerative Colitis & Other Inflammatory Diseases

Andrographolide: Potential Treatment for Ulcerative Colitis & Other Inflammatory Diseases

Andrographolide, a component of the medicinal plant Andrographis paniculata, is a bioactive compound with many biological effects ranging from anti-inflammatory to anti-tumor activities. In a recent study, an andrographolide derivative conjugated with lipoic acid, termed AL-1, was evaluated for its ability to treat a the DSS-Induced Colitis Model of ulcerative colitis (UC). Not only could AL-1 reduce the disease severity, it helped to resolve a key feature that drives the intestinal inflammation characteristic of the DSS-colitis model. This research has promising results for new therapeutics to treat human UC. Read More

Neutrophil Extracellular Traps: not just for catching pathogens.

Neutrophil Extracellular Traps: not just for catching pathogens.

Neutrophil Extracellular Traps (NETs) are exactly as they sound: a web-like structure released from activated neutrophils that can trap and kill pathogens. While the primary function appears to be prolonging the antimicrobial functions of neutrophils, even after cell death, NETs have become a target of research for autoimmune diseases like rheumatoid arthritis and SLE, as well as cancer. Research studying the role of NETs in a variety of disease states could deepen our understanding of inflammatory diseases while providing new avenues for therapeutic development. Read More

Gastrointestinal Microbiota & Reproducibility of Animal Disease Models

Gastrointestinal Microbiota & Reproducibility of Animal Disease Models

Differences in gastrointestinal microbiota contribute to phenotypic variance in experimental disease models, lowering the reproducibility of many preclinical animal model studies. As experts in conducting preclinical animal model experiments, Chondrex, Inc. can help you identify sources of microbial variation in your experiments, as well as make a few recommendations to increase the reproducibility of your experiments. Continue reading to find out how to limit gastrointestinal microbiota variance in your experimental disease model experiments! Read More

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