Dietary supplementation of fructooligosaccharides alleviates enterotoxigenic E. coli-induced disruption of intestinal epithelium in a weaned piglet model

Diarrhea caused by pathogens such as enterotoxigenic E. coli (ETEC) is a serious threat to the health of young animals and human infants. Here, we investigated the protective effect of fructooligosaccharides (FOS) on the intestinal epithelium with ETEC-challenge in a weaned piglet model. Twenty-four weaned piglets were randomly divided into three groups: (1) non-ETEC-challenged control (CON), (2) ETEC-challenged control (ECON), and (3) ETEC challenge + 2.5 g/kg FOS (EFOS). On day 19, the CON pigs were orally infused with sterile culture, while the ECON and EFOS pigs were orally infused with active ETEC (2.5 × 109 colony-forming units). On day 21, pigs were slaughtered to collect venous blood and small intestine. Result showed that the pre-treatment of FOS improved the antioxidant capacity and the integrity of intestinal barrier in the ETEC-challenged pigs without affecting their growth performance. Specifically, comparing with ECON pigs, the level of GSH-Px (glutathione peroxidase) and CAT (catalase) in the plasma and intestinal mucosa of EFOS pigs was increased (P<0.05), and the intestinal barrier marked by ZO-1 and plasmatic DAO was also improved in EFOS pigs. A lower level (P<0.05) of inflammatory cytokines in the intestinal mucosa of EFOS pigs might be involved in the inhibition of TLR4/MYD88/NF-κB pathway. The apoptosis of jejunal cells in EFOS pigs was also lower than that in ECON pigs (P<0.05). Our findings provide convincing evidence of possible prebiotic and protective effect of FOS on the maintenance of intestinal epithelial function under the attack of pathogens.

STUDY OF POST CESAREAN SECTION WOUND INFECTION, COMORBIDITIES AND MICROBIOLOGICAL EPIDEMIOLOGY IN A PRIVATE HOSPITAL, MORAR, M.P. INDIA

INTRODUCTION: Cesarean section stands as one of the most common surgical procedures done worldwide. However, it carries a risk of developing surgical site infection. Surgical site infection can have effect on mother's health in terms of signicant morbidity. AIMS & OBJECTIVE: Present study was undertaken to determine the microbiological prole of surgical site infection among post-partum females who underwent cesarean section. MATERIALS & METHODS: A total of 147 females with surgical site infection were identied in a study conducted in a North Indian teaching care hospital . A semi-structured questionnaire was prepared to know the socio-demographic details along with diagnostic and therapeutic parameters. Mean age of study subjects were 25.61 years with 57.8% cases occurring in age group more than than 25 years. Staphylococcus aureus was the most common identied organism (29.2%) followed by sterile culture (25.2%). Ceftriaxone was the most common given antibiotic (27.9%). Among co-morbidities , anemia and PROM were the most common . resuturing was required in 17.69% of the cases. CONCLUSION: Patient factors , microbiological prole and empirical therapy are a prerequisite for controlling surgical site infection.

Plant Uptake of Lactate-Bound Metals: A Sustainable Alternative to Metal Chlorides

Global agricultural intensification has prompted investigations into biostimulants to enhance plant nutrition and soil ecosystem processes. Metal lactates are an understudied class of organic micronutrient supplement that provide both a labile carbon source and mineral nutrition for plant and microbial growth. To gain a fundamental understanding of plant responses to metal lactates, we employed a series of sterile culture-vessel experiments to compare the uptake and toxicity of five metals (Zn, Mn, Cu, Ni, and Co) supplied in lactate and chloride salt form. Additionally, primary root growth in plate-grown Arabidopsis thaliana seedlings was used to determine optimal concentrations of each metal lactate. Our results suggest that uptake and utilization of metals in wheat (Triticum aestivum L.) when supplied in lactate form is comparable to that of metal chlorides. Metal lactates also have promotional growth effects on A. thaliana seedlings with optimal concentrations identified for Zn (0.5–1.0 µM), Mn (0.5–1.0 µM), Cu (0.5 µM), Ni (1.0 µM), and Co (0.5 µM) lactate. These findings present foundational evidence to support the use of metal lactates as potential crop biostimulants due to their ability to both supply nutrients and stimulate plant growth.

Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells

Adoptive cancer immunotherapy using chimeric antigen receptor (CAR) engineered T-cells holds great promise, although several obstacles hinder the efficient generation of cell products under good manufacturing practice (GMP). Patients are often immune compromised, rendering it challenging to produce sufficient numbers of gene-modified cells. Manufacturing protocols are labour intensive and frequently involve one or more open processing steps, leading to increased risk of contamination. We set out to develop a simplified process to generate autologous gamma retrovirus-transduced T-cells for clinical evaluation in patients with head and neck cancer. T-cells were engineered to co-express a panErbB-specific CAR (T1E28z) and a chimeric cytokine receptor (4αβ) that permits their selective expansion in response to interleukin (IL)-4. Using peripheral blood as starting material, sterile culture procedures were conducted in gas-permeable bags under static conditions. Pre-aliquoted medium and cytokines, bespoke connector devices and sterile welding/sealing were used to maximise the use of closed manufacturing steps. Reproducible IL-4-dependent expansion and enrichment of CAR-engineered T-cells under GMP was achieved, both from patients and healthy donors. We also describe the development and approach taken to validate a panel of monitoring and critical release assays, which provide objective data on cell product quality.

Protocol for the Use of a Novel Bioreactor System for Hydrated Mechanical Testing, Strained Sterile Culture, and Force of Contraction Measurement of Tissue Engineered Muscle Constructs

Bioreactor systems are built as controlled environments for biological processes and utilized in the field of tissue engineering to apply mechanical, spatial, and chemical cues to developing tissue grafts. Often the systems are applied to instruct differentiation and maturation of the cells grown inside. Perhaps the most obvious targets for strain and compression-based bioreactors are mechanically active tissues, as it is hypothesized that biomimetic mechanical environments instruct immature cells to form differentiated tissues. One such tissue, skeletal muscle, has been identified as a key candidate for strain application due to the close structure-function relationship of myofibers. Here we detail the multiple uses of a custom-built bioreactor system in combination with electrospun fibrin microfibers for muscle tissue engineering. Outlined below are the methods used in the system to test the mechanical properties of hydrogel-based scaffolds in an aqueous environment, including Young’s modulus and poroelasticity. Additionally, we demonstrate the application of tensile strain to sterile cell cultures grown on electrospun scaffolds and perform end-point testing of tissue contractility with the addition of an electrode.

INFLUENCE OF THE STERILIZATION METHOD AND THE TIME OF INTRODUCTION INTO IN VITRO CULTURE ON THE VIABILITY OF EXPLANTS OF THE CLONAL APPLE STOCK 54-118

The stage of introducing explants into a sterile culture is difficult in the technology of clonal micropropagation of plants. The article shows the possible ways of sterilization and the introduction terms of explants of the clonal apple stocks 54-118 into in vitro culture in order to reduce planting infection and increase the yield of sterile viable explants. The best time for introducing clonal apple stocks 54-118 into sterile in vitro culture is the period of active shoot growth. Sterilization of explants with ethyl alcohol (70,0 %, 1 min) in combination with hydrogen peroxide (33,0 %) for 7 minutes and ethyl alcohol (70,0 %, 1 min) in combination with diacide (0,1 %) within 6 minutes contributed to the production of 63,0 % and 60,0 % of viable sterile explants.

Combined testing approach to evaluate the antifungal efficiency of clove (Eugenia caryophyllata) essential oil for potential application in wood conservation

The efficiency of clove (Eugenia caryophyllata) essential oil (C-EO) for the curative antifungal treatment of historic wood was investigated in comparison with two classical biocide products: a boron-based preservative (Diffusit S) and a formulation containing quaternary ammonium salts and isothiazole (Biotin T). A combined approach was adopted that consisted of implementation of C-EO in a practical case study on a degraded beech (Fagus sylvatica) wood artifact and evaluation of the treatment efficacy via an original laboratory mycological test. Small samples, extracted from the degraded wood material before and after curative treatments, were placed as inoculum on sterile culture medium and incubated for periodic monitoring of the emerging fungal growth for 140 d. Direct observation was supplemented with digital quantification of the fungal coverage area via ImageJ software and calculation of the absolute and relative indices of fungal development reduction. The results indicated that the C-EO solutions at both tested concentrations (10%, 5%) were more efficient than the considered reference products at similar concentrations (Diffusit S (10%) and Biotin T (5%)) for curative antifungal treatment. However, none of the treatments applied entirely prevented reactivation of the severe and complex fungal attack, which was highlighted by the mycological tests conducted on the control samples.

Cardiopulmonary Lesions in Sheep Produced by Experimental Acute Clostridium Perfringens Type D Enterotoxemia

Enterotoxemia caused by Clostridium perfringens type D is one of the most prevalent clostridial diseases of sheep. The lesions of the acute form of this disease, particularly the cerebral lesions, are well characterized; however, detailed descriptions of the cardiac and pulmonary lesions are lacking. Here we describe cardiopulmonary lesions in experimental acute type D enterotoxemia in sheep and determine the role of epsilon toxin (ETX) in the development of these lesions. Four groups of 6 sheep were intraduodenally inoculated with either a wild-type C. perfringens type D strain; its etx knockout mutant, which is unable to produce ETX; the etx mutant complemented with the wild-type etx gene, which regains the ETX toxigenic ability; or sterile culture medium as a control. All sheep were subjected to postmortem examination within 24 hours of inoculation. Lesion scores were compared between groups for pulmonary edema; hydrothorax; ascites; hydropericardium; endocardial, myocardial and epicardial hemorrhages; microscopic lesions of acute myocardial degeneration and necrosis; and myocardial, endocardial, and epicardial edema, hemorrhage, and inflammation. Only sheep inoculated with the wild-type and complemented ETX-toxigenic bacterial strains developed cardiopulmonary lesions, which were present in varying degrees of severity and proportions. These lesions were not present in sheep inoculated with the etx mutant or in the negative control. We conclude that severe acute cardiopulmonary lesions in sheep with experimental enterotoxemia are associated with the capacity of the strains to produce ETX. These changes are likely contributors to the clinical signs and even death of affected animals.

Clinical significance of DJ stent culture in patients with indwelling ureteral stents prior to endourological intervention

Background: Ureteral stents are placed to relieve ureteral obstruction in various endourological surgery. As soon as it is inserted it is rapidly colonized by host of microorganism. Culture and sensitivity pattern of these growing microorganism on the stent may provide us an insight to treat postoperative urosepsis in high risk patients with indwelling DJ stents. Materials and methods: Prospective study between September 2018 and August 2019 with enrollment of 100 patients to determine the ability of urine culture (UC) to predict stent culture (SC) status, to identify associated infectious pathogens with their antibiotic sensitivity patterns and to evaluate the association between SC and post-endourological intervention sepsis. Results: Colonization of bacteria on stent culture (SC) was identified in 53 out of 100 patients (53%). Most of the organism grown on SC were multidrug resistant and they were resistant to most commonly prescribed antibiotics like fluroquinolones, cephalosporins, and aminoglycosides .These organisms were only sensitive to newer antibiotics. Indwelling time of ureteral stent was directly proportional with SC positivity ( p < 0.05). Association of Urine culture at DJ Removal and DJ stent culture was not statistically significant. Conclusion: Very low rates of colonization within 6 weeks after the insertion of stents. Sterile culture of urine does not rule out colonization of the stent. Most of the urinary pathogens as well as the organisms grown on DJ stent are resistant to fluroquinolones and cephalosporins.