A distinct bacterial dysbiosis associated skin inflammation in ovine footrot

Ovine footrot is a highly prevalent bacterial disease caused by Dichelobacter nodosus and characterised by the separation of the hoof horn from the underlying skin. The role of innate immune molecules and other bacterial communities in the development of footrot lesions remains unclear. This study shows a significant association between the high expression of IL1β and high D. nodosus load in footrot samples. Investigation of the microbial population identified distinct bacterial populations in the different disease stages and also depending on the level of inflammation. Treponema (34%), Mycoplasma (29%) and Porphyromonas (15%) were the most abundant genera associated with high levels of inflammation in footrot. In contrast, Acinetobacter (25%), Corynebacteria (17%) and Flavobacterium (17%) were the most abundant genera associated with high levels of inflammation in healthy feet. This demonstrates for the first time there is a distinct microbial community associated with footrot and high cytokine expression

Intraperitoneal drain placement and outcomes after elective colorectal surgery: international matched, prospective, cohort study

Despite current guidelines, intraperitoneal drain placement after elective colorectal surgery remains widespread. Drains were not associated with earlier detection of intraperitoneal collections, but were associated with prolonged hospital stay and increased risk of surgical-site infections.Background Many surgeons routinely place intraperitoneal drains after elective colorectal surgery. However, enhanced recovery after surgery guidelines recommend against their routine use owing to a lack of clear clinical benefit. This study aimed to describe international variation in intraperitoneal drain placement and the safety of this practice. Methods COMPASS (COMPlicAted intra-abdominal collectionS after colorectal Surgery) was a prospective, international, cohort study which enrolled consecutive adults undergoing elective colorectal surgery (February to March 2020). The primary outcome was the rate of intraperitoneal drain placement. Secondary outcomes included: rate and time to diagnosis of postoperative intraperitoneal collections; rate of surgical site infections (SSIs); time to discharge; and 30-day major postoperative complications (Clavien-Dindo grade at least III). After propensity score matching, multivariable logistic regression and Cox proportional hazards regression were used to estimate the independent association of the secondary outcomes with drain placement. Results Overall, 1805 patients from 22 countries were included (798 women, 44.2 per cent; median age 67.0 years). The drain insertion rate was 51.9 per cent (937 patients). After matching, drains were not associated with reduced rates (odds ratio (OR) 1.33, 95 per cent c.i. 0.79 to 2.23; P = 0.287) or earlier detection (hazard ratio (HR) 0.87, 0.33 to 2.31; P = 0.780) of collections. Although not associated with worse major postoperative complications (OR 1.09, 0.68 to 1.75; P = 0.709), drains were associated with delayed hospital discharge (HR 0.58, 0.52 to 0.66; P < 0.001) and an increased risk of SSIs (OR 2.47, 1.50 to 4.05; P < 0.001). Conclusion Intraperitoneal drain placement after elective colorectal surgery is not associated with earlier detection of postoperative collections, but prolongs hospital stay and increases SSI risk

Expression of T399I and D299G polymorphisms of TLR4 gene in colorectal cancer cell line by flowcytometery

Background and Objective: Toll-like receptors (TLRs) have been discovered as the most important receptors in innate immunity. One of the most important TLRs is TLR4, the key receptor for the LPS component of gram-negative bacteria. Two polymorphisms, D299G (rs4986790) and T399I (rs4986791), in TLR4 gene are associated with a decreased response to LPS. This study was done to estimate the expression of different polymorphisms of TLR4 gene in colorectal cancer cell line by flowcytometery. Materials and Methods: In this laboratory study, the HCT116 cells were transfected with plasmids containing different variants of TLR4 gene including Flag-tagged-TLR4 wild type, flag-tagged D299G and T399I Using TurboFect transfection reagent. Transfection efficiency was evaluated by GFP plasmid. Expression of different variants of TLR4 was assessed in transfected cells by flowcytometery. Data were analyzed using SPSS-11.5 and chi-square test. Results: TLR4 was detected on HT29 and CaCo2 cell lines at low levels. HCT116 cells did not express detectable amounts of TLR4 by flowcytometery prior to transfection. Gene transfer efficiency for GFP plasmid was about 80% in HCT116 cells by flowcytometery and microscopic analysis. TLR4 expression and LPS responsiveness significantly was higher in HCT116 cells which were transfected with wild type TLR4 gene compared to non-transfected and mutant transfected cells (P<0.05). Conclusion: Lower expression of TLR4 on cells with mutant TLR4 showed that these polymorphisms affect on expression patterns of TLR4 on colon cancer cells

Advances in targeted and immunobased therapies for colorectal cancer in the genomic era

Targeted therapies require information on specific defective signaling pathways or mutations. Advances in genomic technologies and cell biology have led to identification of new therapeutic targets associated with signal-transduction pathways. Survival times of patients with colorectal cancer (CRC) can be extended with combinations of conventional cytotoxic agents and targeted therapies. Targeting EGFR- and VEGFR-signaling systems has been the major focus for treatment of metastatic CRC. However, there are still limitations in their clinical application, and new and better drug combinations are needed. This review provides information on EGFR and VEGF inhibitors, new therapeutic agents in the pipeline targeting EGFR and VEGFR pathways, and those targeting other signal-transduction pathways, such as MET, IGF1R, MEK, PI3K, Wnt, Notch, Hedgehog, and death-receptor signaling pathways for treatment of metastatic CRC. Additionally, multitargeted approaches in combination therapies targeting negative-feedback loops, compensatory networks, and cross talk between pathways are highlighted. Then, immunobased strategies to enhance antitumor immunity using specific monoclonal antibodies, such as the immune-checkpoint inhibitors anti-CTLA4 and anti-PD1, as well as the challenges that need to be overcome for increased efficacy of targeted therapies, including drug resistance, predictive markers of response, tumor subtypes, and cancer stem cells, are covered. The review concludes with a brief insight into the applications of next-generation sequencing, expression profiling for tumor subtyping, and the exciting progress made in in silico predictive analysis in the development of a prescription strategy for cancer therapy

Cytokine gene expression in sheep following experimental infection with various strains of Corynebacterium pseudotuberculosis differing in virulence

International audienc

Metabolites from Actinomyces strain H6552 extract inhibit transforming growth factor-mediated pulmonary fibrosis

Purpose: To evaluate the effects of H6552 extract in inhibiting transforming growth factor (TGF)- mediated pulmonary fibrosis in vitro and in vivo.Methods: Maximum-nontoxic dose (MNTD) of Actinomyces H6552 extract was determined using 3- (4,5-dimethylthiazol-2-yl)-2,5-diphhenyltetrazolium bromide (MTT) assay. Effect of the extract on IMR- 90 lung fibroblasts proliferation was determined by calculating the population doubling time (PDT). Collagen gel contraction assay was carried out to determine cell contractility while α-smooth muscle actin (SMA) level in cells was evaluated by quantitative real-time polymerase chain reaction (PCR) and immunostaining methods. A bleomycin-induced ICR mouse model was used in the study to determine the effect of the extract in vivo. The animals received treatments in two regimes: early treatment in which treatment was given on Day 0 and delayed treatment with treatment on Days 5 and 10. The animals were sacrificed on Day 14 and the lungs removed for histopathological assessment.Results: The MNTD of the H6552 extract was 1625 ± 459.62 μg/ml. H6552 extract significantly reduced TGF- β-mediated cell proliferation, gel contraction and α-SMA expression. PDT was increased up to 83.84 % in the treated cells. Gel contraction was inhibited by the addition of 1000 μg/ml of H6552 extract. Immunostaining results revealed negligible α-SMA antibody staining after H6552 extract treatment at 500 μg/ml. The extract also inhibited lung injury (54 % reduction in Ashcroft score) when early treatment was provided. Delayed treatment with the extract did not show any significant changes in the animals.Conclusion: H6552 extract inhibited TGF-β-induced pulmonary fibrosis and elucidation of its bioactive metabolites may yield a potential agent to treat the disease.Keywords: Actinomyces H6552, Transforming growth factor-β, Cell contractility, α-Smooth muscle actin, Pulmonary fibrosi