Accumulation, Chronicity, and Induction of Oxidative Stress Regulating Genes Through Allium cepa L. Functionalized Silver Nanoparticles in Freshwater Common Carp (Cyprinus carpio).

Green evolutionary products such as biologically fabricated nanoparticles (NPs) pose a hazard to aquatic creatures. Herein, biogenic silver nanoparticles (AgNPs) were synthesized by the reaction between ionic silver (AgNO3) and aqueous onion peel extract (Allium cepa L). The synthesized biogenic AgNPs were characterized with UV-Visible spectrophotometer, XRD, FT-IR, and TEM with EDS analysis; then, their toxicity was assessed on common carp fish (Cyprinus carpio) using biomarkers of haematological alterations, oxidative stress, histological changes, differential gene expression patterns, and bioaccumulation. The 96 h lethal toxicity was analysed with various concentrations (2, 4, 6, 8, and 10 mg/l) of biogenic AgNPs. Based on 96 h LC50, sublethal concentrations (1/15th, 1/10th, and 1/5th) were given to C. carpio for 28 days. At the end of experiment, the bioaccumulations of Ag content were accumulated mainly in the gills, followed by the liver and muscle. At an interval of 7 days, the haematological alterations showed significance (p < 0.05) and elevation of antioxidant defence mechanism reveals the toxicity of biogenic synthesized AgNPs. Adverse effects on oxidative stress were probably related to the histopathological damage of its vital organs like gill, liver, and muscle. Finally, the fish treated with biogenic synthesized AgNPs were significantly (p < 0.05) downregulates the oxidative stress genes such as Cu-Zn SOD, CAT, GPx1a, GST-α, CYP1A, and Nrf-2 expression patterns. The present study provides evidence of biogenic synthesized AgNPs influence on the aquatic life through induction of oxidative stress.

Characterization of Intestinal Mesenchymal Stromal Cells From Patients With Inflammatory Bowel Disease for Autologous Cell Therapy.

Therapy with mesenchymal stromal cells (MSCs) has shown promise in inflammatory bowel disease-leveraging their immunosuppressive and regenerative properties. However, the potential immunogenic complications of allogenic MSCs sourced from different tissues raise concern. Thus, we assessed the fitness and functionality of autologous intestinal MSCs as a potential platform for cellular therapy. Mucosal biopsy-derived MSCs from Crohn's disease (n = 11), ulcerative colitis (n = 12), and controls (n = 14) were analyzed by microscopy and flow cytometry for doubling-time, morphology, differentiation potential, and immunophenotype. Gene expression, cell-subtype composition, along with surface marker and secretome changes after IFN-γ priming were measured by bulk and single-cell RNA sequencing coupled with a 30-plex Luminex panel. MSCs expanded ex vivo demonstrate canonical MSC markers, similar growth kinetics, and tripotency regardless of the patient phenotype. Global transcription patterns were similar at baseline though inflammatory bowel disease (IBD) rectal MSCs showed changes in select immunomodulatory genes. IFN-γ priming resulted in upregulation of shared immunoregulatory genes (particularly in PD-1 signaling) and overrode the transcriptional differences observed at baseline. Furthermore, MSCs secrete key immunomodulatory molecules at baseline and in response to IFN-γ including CXCL10, CXCL9, and MCP-1. Overall, MSCs from IBD patients have normal transcriptional and immunomodulatory properties with therapeutic potential and can be sufficiently expanded.

Ileal Derived Organoids From Crohn's Disease Patients Show Unique Transcriptomic and Secretomic Signatures.

BACKGROUND & AIMS: We used patient-derived organoids (PDOs) to study the epithelial-specific transcriptional and secretome signatures of the ileum during Crohn's disease (CD) with varying phenotypes to screen for disease profiles and potential druggable targets. METHODS: RNA sequencing was performed on isolated intestinal crypts and 3-week-old PDOs derived from ileal biopsies of CD patients (n = 8 B1, inflammatory; n = 8 B2, stricturing disease) and non-inflammatory bowel disease (IBD) controls (n = 13). Differentially expressed (DE) genes were identified by comparing CD vs control, B1 vs B2, and inflamed vs non-inflamed. DE genes were used for computational screening to find candidate small molecules that could potentially reverse B1and B2 gene signatures. The secretome of a second cohort (n = 6 non-IBD controls, n = 7 CD, 5 non-inflamed, 2 inflamed) was tested by Luminex using cultured organoid conditioned medium. RESULTS: We found 90% similarity in both the identity and abundance of protein coding genes between PDOs and intestinal crypts (15,554 transcripts of 19,900 genes). DE analysis identified 814 genes among disease group (CD vs non-IBD control), 470 genes different between the CD phenotypes, and 5 false discovery rate correction significant genes between inflamed and non-inflamed CD. The PDOs showed both similarity and diversity in the levels and types of soluble cytokines and growth factors they released. Perturbagen analysis revealed potential candidate compounds to reverse B2 disease phenotype to B1 in PDOs. CONCLUSIONS: PDOs are similar at the transcriptome level with the in vivo epithelium and retain disease-specific gene expression for which we have identified secretome products, druggable targets, and corresponding pharmacologic agents. Targeting the epithelium could reverse a stricturing phenotype and improve outcomes.

Clinically important microbial diversity and its antibiotic resistance pattern towards various drugs.

BACKGROUND: Increased use of antibiotics in poultry leads to the development of antimicrobial resistance among the commensal bacterium of broiler chickens. OBJECTIVE: In this study, we aimed at studying the effect of periodic administration of therapeutic antibiotics against the bacterial diversity in poultry litters collected from broiler chickens. METHODS: Poultry litters were collected randomly at regular intervals after administration of antibiotics (1st, 12th and 22nd day) to the chicken. Bedding material without litters served as control. Phenotypic observations showed that there is a difference in the bacterial richness isolated at regular intervals. A total of 32 bacteria were isolated from poultry letters and are grouped into ten different genus. Isolated bacterial species were further confirmed by16S rRNA sequencing. RESULTS: Antibiotic susceptibility profile of isolated bacterial species exhibited strong resistance towards 13 selected antibiotics. These results substantiate that administration of antibiotics leads to the alterations in bacterial diversity and development of antimicrobial resistance among the commensal bacteria of poultry litter. CONCLUSION: This high selection pressure of therapeutic antibiotics may lead to species selection and development of antibiotic resistance among bacterial population. Development of such species selection may access the human and other organisms via food chain and can cause severe health defects.