Prevalence of Listeria species in some foods and their rapid identification

Purpose: To investigate the occurrence of Listeria spp., (particularly L. monocytogenes), in different foods and to compare diagnostic tools for their identification at species level.Methods: Samples of high protein foods such as raw meats and meat products and including beef products, chicken, fish and camel milk were analysed for the presence of Listeria spp. The isolates were characterised by morphological and cultural analyses, and confirmed isolates were identified by protein profiling and verified using API Listeria system. Protein profiling by SDS-PAGE was also used to identify Listeria spp.Results: Out of 40 meat samples, 14 (35 %) samples were contaminated with Listeria spp., with the highest incidence (50 %) occurring in raw beef products and raw chicken. Protein profiling by SDSPAGE was used to identify Listeria spp. The results were verified with API Listeria system. Approximately 25 % of the identified isolates were Listeria seeligeri, Listeria welshimeri, and Listeria grayi (three positive samples), while 16.66 % of the isolates were Listeria monocytogenes (two positive samples); 16.6 % of the isolates were Listeria innocua (two positive samples), while 8.3 % of the isolates were Listeria ivanovii (one positive sample).Conclusion: High protein foods contain different types of Listeria species; whole-cell protein profiles and API Listeria system can help in the identification of Listeria at the species level.Keywords: Listeria spp, High protein food, Api Listeria, Protein profil

Prevalence of hepatitis C virus in patients with tuberculosis and its impact in the incidence of anti-tuberculosis drugs induced hepatotoxicity

AbstractBackgroundThe prevalence of hepatitis C virus (HCV) infection among patients with tuberculosis (TB) has not been extensively investigated, and very limited data on rates of HCV co-infection among patients with TB exists. Hepatotoxicity is the major adverse effect of three of the first line anti-TB agents: isoniazid (INH), rifampin (RIF), and pyrazinamide (PZA). Chronic liver disease raises a risk of hepatotoxicity during anti-tuberculosis treatment, up to three to five times more than TB patients who do not have viral infection.AimTo assess the prevalence of HCV infection in patients with tuberculosis and its impact in the incidence of anti-tuberculosis drug induced hepatotoxicity (DIH).Subjects and methodsThe prevalence of HCV in patients with newly diagnosed pulmonary or extrapulmonary tuberculosis was estimated using polymerase chain reaction (PCR). Then patients were classified into 2 groups: group I (patients with HCV-TB coinfections) and group II (HCV-seronegative tuberculous patients). Baseline and monthly measuring liver transaminases was done before and following the start of 1st line anti-tuberculosis therapy.ResultsThe prevalence of HCV in patients with TB was 17.02%. Regarding DIH, in group I; 6 (40%) cases showed transient transaminase elevations and 6 (40%) cases developed DIH. In group II; 11 (20.75%) cases developed transient transaminase elevations and only 2 (3.78%) cases developed DIH, and there was a highly significant difference (<0.01) between both groups. Regarding the severity of DIH, in group I; 4 cases were mild, one case was moderate and one case was severe. While in group II, no cases was with severe DIH. The risk factors for developing DIH during anti-tuberculosis therapy were; age ⩾40, high baselines transaminases, ALP and total bilirubin, and low BMI. Most cases of DIH occurred during the 1st 4weeks of starting anti-tuberculosis therapy (66.7% and 50% in group I and group II, respectively).ConclusionsTuberculosis and hepatitis C virus co-infection is common, and elevation of liver functions during anti-tuberculosis therapy is not uncommon. HCV-positive patients with tuberculosis should be closely monitored during treatment especially if they had elevated baseline liver functions, old age and with low BMI. Monitoring should include the whole period of treatment, especially the 1st 2months

Sound analysis to predict the growth of Turkeys

Protocols for manual weighing of turkeys are not practical on turkey farms because of the large body sizes, heavy weights and flighty nature of turkeys. The sounds turkeys make may be a proxy for bird weights, but the relationship between turkey sounds and bird weights has not been studied. The aim of this study was to correlate peak frequency (PF) of vocalization with the age and weight of the bird and examine the possibility using PF to predict the weight of turkeys. The study consisted of four trials in Egypt. Sounds of birds and their weights were recorded for 11 days during the growth period in each trial. A total 2200 sounds were manually analyzed and labelled by extracting individual and general sounds on the basis of the amplitude and frequency of the sound signal. The PF of vocalizations in each trial, as well as in pooled trails, were evaluated to determine the relationship between PF and the age and weight of the turkey. PF exhibited a highly significant negative correlation with the weight and age of the turkeys showing that PF of vocalizations can be used for predicting the weight of turkeys. Further studies are necessary to refine the procedure

Calculation the Cross Sections for64Cu(n,p)64Ni Reaction By Reciprocity Theory

In this study intermediate elements 64Ni , 64Cu for 64Ni (p,n)64Cu reaction with proton energy from (1.0) MeV to (132) MeV with threshold energy (2.496) MeV are used according to the available data of reaction cross sections. We calculated the cross sections for 64Cu(n,p)64Ni reaction by application in nuclear technology (reciprocity theory). In reciprocity theory we derive the mathematical formula for 64Cu(n,p)64Ni and we deduced high probability to produced 64Ni because it is very important such as it used in technology field .The evaluated cross sections as a functions of neutron energy between (En =0.504MeV) to (En=129.506MeV) of (0.0106barn) (0.254barn) respectively and statistical factor (gp,n=1 and gn,p=1/3)

Cross Sections Calculations of 33S(n,α)30Si reaction by using the inverse reaction for the first exited state

In this study light elements 30Si , 33S for 30Si(α,n)33S reaction as well as α-particle energy from (5.001) MeV to (6.284) MeV are used according to the available data of reaction cross sections with threshold energy (3.959MeV). The more recent cross sections data of 30Si(α,n)33S reaction is reproduced in fine steps in the specified energy range , as well as cross section (α,n) values were derived from the published data of (n,α) as a function of energy in the same fine energy steps by using the principle of inverse reaction . This calculations involves the first exited state of 30Si , 33S in the reactions 30Si(α,n)33S and 33S(n,α)30Si . These data are listing , plotted and dissection

Immune checkpoint inhibitor-induced colitis is mediated by polyfunctional lymphocytes and is dependent on the IL23/IFNg axis

Immune checkpoint inhibitors (CPIs) have revolutionised cancer treatment, with previously untreatable disease now amenable to potential cure. Combination regimens of anti-CTLA-4 and anti-PD-1 show enhanced efficacy but are prone to off-target immune-mediated tissue injury, particularly at the barrier surfaces. CPI-induced colitis is a common and serious complication. To probe the impact of immune checkpoints on intestinal homeostasis, mice were challenged with combination anti-CTLA-4/anti-PD-1 immunotherapy and manipulation of the intestinal microbiota. Colonic immune responses were profiled using bulk and single-cell RNA-sequencing and flow cytometry. CPI-colitis was dependent on the composition of the intestinal microbiota and was characterized by remodelling of mucosal lymphocytes with induction of polyfunctional lymphocyte responses characterized by increased expression of interferon-γ (IFNγ), other pro-inflammatory cytokines/chemokines (Il22, Il17a Ccl3, Ccl4 and Ccl9), cytotoxicity molecules (Gzmb, Gzma, Prf1, Nkg7) and the chemokine receptor Cxcr6. In comparison with mucosal lymphocytes in the steady state, polyfunctional lymphocytes from both CD4+ and CD8+ lineages upregulated costimulatory molecules and checkpoint molecules in CPI-colitis, indicating that these cells are tightly regulated. CPI-colitis was attenuated following depletion of effector lymphocytes or following blockade of the IL23/IFNγ axis. This study provides new mechanistic insights into CPI-colitis, identifying polyfunctional, cytotoxic lymphocytes as key mediators of disease. Therapeutic targeting of their effector response or regulatory networks, including the IL23/IFNγ axis likely holds the key to preventing and reversing CPI-colitis

Effectiveness and safety of vedolizumab in inflammatory bowel disease patients aged 60 and over: an observational multicenter UK experience.

BACKGROUND: The GEMINI trials established the efficacy of vedolizumab in moderate-to-severe inflammatory bowel disease (IBD) and demonstrated a favorable safety profile, suggesting it may be advantageous in older patients at greater risk of treatment-related complications. However, there is a paucity of data exploring the outcomes of vedolizumab in this group. Our objective was to determine the clinical effectiveness and safety of vedolizumab in older IBD patients within a real-world multicenter UK cohort. METHODS: A retrospective review of electronic records across 6 UK hospitals was undertaken to evaluate the clinical effectiveness and safety outcomes of vedolizumab in IBD patients aged ≥60 at start of therapy. Rates of clinical response, remission and corticosteroid-free remission were assessed at weeks 14 and 52, using validated clinical indices, and were compared to historical controls from real-world vedolizumab-treated cohorts unstratified by age. RESULTS: Of 74 patients aged 60 years or above (median 66 years), 48 were included in our effectiveness analysis (29 ulcerative colitis, 19 Crohn's disease). Rates of clinical response, remission and corticosteroid-free remission at week 14 were 64%, 48% and 30%, respectively. By week 52, the rates of clinical response, remission, and corticosteroid-free remission were 52%, 38%, and 32%, respectively. Six (8%) patients experienced adverse effects. Effectiveness and safety outcomes were comparable to those of age-unstratified vedolizumab-treated cohorts. CONCLUSION: Our 1-year outcome data suggests that vedolizumab is safe and effective in older IBD patients and broadly comparable to cohorts unselected by age

CRISPR-enhanced human adipocyte \u27browning\u27 as cell therapy for metabolic disease [preprint]

Obesity and type 2 diabetes (T2D) are associated with poor tissue responses to insulin [1,2], disturbances in glucose and lipid fluxes [3-5] and comorbidities including steatohepatitis [6] and cardiovascular disease [7,8]. Despite extensive efforts at prevention and treatment [9,10], diabetes afflicts over 400 million people worldwide [11]. Whole body metabolism is regulated by adipose tissue depots [12-14], which include both lipid-storing white adipocytes and less abundant \u27brown\u27 and \u27brite/beige\u27 adipocytes that express thermogenic uncoupling protein UCP1 and secrete factors favorable to metabolic health [15-18]. Application of clustered regularly interspaced short palindromic repeats (CRISPR) gene editing [19,20] to enhance \u27browning\u27 of white adipose tissue is an attractive therapeutic approach to T2D. However, the problems of cell-selective delivery, immunogenicity of CRISPR reagents and long term stability of the modified adipocytes are formidable. To overcome these issues, we developed methods that deliver complexes of SpyCas9 protein and sgRNA ex vivo to disrupt the thermogenesis suppressor gene NRIP1 [21,22] with near 100% efficiency in human or mouse adipocytes. NRIP1 gene disruption at discrete loci strongly ablated NRIP1 protein and upregulated expression of UCP1 and beneficial secreted factors, while residual Cas9 protein and sgRNA were rapidly degraded. Implantation of the CRISPR-enhanced human or mouse brown-like adipocytes into high fat diet fed mice decreased adiposity and liver triglycerides while enhancing glucose tolerance compared to mice implanted with unmodified adipocytes. These findings advance a therapeutic strategy to improve metabolic homeostasis through CRISPR-based genetic modification of human adipocytes without exposure of the recipient to immunogenic Cas9 or delivery vectors

Clinical outcomes of patients with corticosteroid refractory immune checkpoint inhibitor induced enterocolitis treated with infliximab

Introduction Immune Checkpoint Inhibitors (CPI) have changed the treatment landscape for many cancers, but also cause severe inflammatory side effects including enterocolitis. CPI-induced enterocolitis is treated empirically with corticosteroids, and infliximab (IFX) is used in corticosteroid-refractory cases. However, robust outcome data for these patients are scarce. Methods We conducted a multi-centre (six cancer centres), cohort study of outcomes in patients treated with IFX for corticosteroid-refractory CPI-induced enterocolitis between 2007 and 2020. The primary outcome was corticosteroid-free clinical remission (CFCR) with CTCAE grade 0 for diarrhoea at 12 weeks after IFX initiation. We also assessed cancer outcomes at one year using RECIST criteria. Results 127 patients (73 male; median age 59 years) were treated with IFX for corticosteroid-refractory CPI-induced enterocolitis. Ninety-six (75.6%) patients had diarrhoea CTCAE grade >2 and 115 (90.6%) required hospitalisation for colitis. CFCR was 41.2% at 12 weeks and 50.9% at 26 weeks. In multivariable logistical regression, IFX-resistant enterocolitis was associated with rectal bleeding (OR 0.19; 95% CI 0.04-0.80; p=0.03) and absence of colonic crypt abscesses (OR 2.16; 95% CI 1.13-8.05; p=0.03). Cancer non-progression was significantly more common in patients with IFX-resistant enterocolitis (64.4%) as compared to patients with IFX-responsive enterocolitis (37.5%; p=0.013). Conclusion This is the largest study to date reporting outcomes of IFX therapy in patients with corticosteroid-refractory CPI-induced enterocolitis. Utilizing pre-defined robust endpoints, we have demonstrated that fewer than half of patients achieved CFCR. Our data also indicate that cancer outcomes may be better in patients developing prolonged and severe inflammatory side effects of CPI-therapy

The gut microbiota and metabolome are associated with diminished COVID-19 vaccine-induced antibody responses in immunosuppressed inflammatory bowel disease patients

Background: Patients with inflammatory bowel disease (IBD) treated with anti-TNF therapy exhibit attenuated humoral immune responses to vaccination against SARS-CoV-2. The gut microbiota and its functional metabolic output, which are perturbed in IBD, play an important role in shaping host immune responses. We explored whether the gut microbiota and metabolome could explain variation in anti-SARS-CoV-2 vaccination responses in immunosuppressed IBD patients. Methods: Faecal and serum samples were prospectively collected from infliximab-treated patients with IBD in the CLARITY-IBD study undergoing vaccination against SARS-CoV-2. Antibody responses were measured following two doses of either ChAdOx1 nCoV-19 or BNT162b2 vaccine. Patients were classified as having responses above or below the geometric mean of the wider CLARITY-IBD cohort. 16S rRNA gene amplicon sequencing, nuclear magnetic resonance (NMR) spectroscopy and bile acid profiling with ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) were performed on faecal samples. Univariate, multivariable and correlation analyses were performed to determine gut microbial and metabolomic predictors of response to vaccination. Findings: Forty-three infliximab-treated patients with IBD were recruited (30 Crohn's disease, 12 ulcerative colitis, 1 IBD-unclassified; 26 with concomitant thiopurine therapy). Eight patients had evidence of prior SARS-CoV-2 infection. Seventeen patients (39.5%) had a serological response below the geometric mean. Gut microbiota diversity was lower in below average responders (p = 0.037). Bilophila abundance was associated with better serological response, while Streptococcus was associated with poorer response. The faecal metabolome was distinct between above and below average responders (OPLS-DA R2X 0.25, R2Y 0.26, Q2 0.15; CV-ANOVA p = 0.038). Trimethylamine, isobutyrate and omega-muricholic acid were associated with better response, while succinate, phenylalanine, taurolithocholate and taurodeoxycholate were associated with poorer response. Interpretation: Our data suggest that there is an association between the gut microbiota and variable serological response to vaccination against SARS-CoV-2 in immunocompromised patients. Microbial metabolites including trimethylamine may be important in mitigating anti-TNF-induced attenuation of the immune response. Funding: JLA is the recipient of an NIHR Academic Clinical Lectureship (CL-2019-21-502), funded by Imperial College London and The Joyce and Norman Freed Charitable Trust. BHM is the recipient of an NIHR Academic Clinical Lectureship (CL-2019-21-002). The Division of Digestive Diseases at Imperial College London receives financial and infrastructure support from the NIHR Imperial Biomedical Research Centre (BRC) based at Imperial College Healthcare NHS Trust and Imperial College London. Metabolomics studies were performed at the MRC-NIHR National Phenome Centre at Imperial College London; this work was supported by the Medical Research Council (MRC), the National Institute of Health Research (NIHR) (grant number MC_PC_12025) and infrastructure support was provided by the NIHR Imperial Biomedical Research Centre (BRC). The NIHR Exeter Clinical Research Facility is a partnership between the University of Exeter Medical School College of Medicine and Health, and Royal Devon and Exeter NHS Foundation Trust. This project is supported by the National Institute for Health Research (NIHR) Exeter Clinical Research Facility. The views expressed are those of the authors and not necessarily those of the NIHR or the UK Department of Health and Social Care