Restriction of growth and biofilm formation of ESKAPE pathogens by caprine gut-derived probiotic bacteria.

The accelerated rise in antimicrobial resistance (AMR) poses a significant global health risk, necessitating the exploration of alternative strategies to combat pathogenic infections. Biofilm-related infections that are unresponsive to standard antibiotics often require the use of higher-order antimicrobials with toxic side effects and the potential to disrupt the microbiome. Probiotic therapy, with its diverse benefits and inherent safety, is emerging as a promising approach to prevent and treat various infections, and as an alternative to antibiotic therapy. In this study, we isolated novel probiotic bacteria from the gut of domestic goats (Capra hircus) and evaluated their antimicrobial and anti-biofilm activities against the 'ESKAPE' group of pathogens. We performed comprehensive microbiological, biochemical, and molecular characterizations, including analysis of the 16S-rRNA gene V1-V3 region and the 16S-23S ISR region, on 20 caprine gut-derived lactic acid bacteria (LAB). Among these, six selected Lactobacillus isolates demonstrated substantial biofilm formation under anaerobic conditions and exhibited robust cell surface hydrophobicity and autoaggregation, and epithelial cell adhesion properties highlighting their superior enteric colonization capability. Notably, these Lactobacillus isolates exhibited broad-spectrum growth inhibitory and anti-biofilm properties against 'ESKAPE' pathogens. Additionally, the Lactobacillus isolates were susceptible to antibiotics listed by the European Food Safety Authority (EFSA) within the prescribed Minimum Inhibitory Concentration limits, suggesting their safety as feed additives. The remarkable probiotic characteristics exhibited by the caprine gut-derived Lactobacillus isolates in this study strongly endorse their potential as compelling alternatives to antibiotics and direct-fed microbial (DFM) feed supplements in the livestock industry, addressing the escalating need for antibiotic-free animal products.

Early Definitive Diagnosis and Management of Incidental Neuroendocrine Tumors Found on Gastrointestinal Endoscopy.

Neuroendocrine tumors (NETs) are tumors that originate from neuroendocrine cells and can be found throughout the body but are most commonly seen in the gastrointestinal tract, pancreas, and lungs. There is an increase in the diagnosis of NETs due to advances in diagnostic modalities. Although mucosal tumors are easily visualized on upper GI endoscopic imaging, neuroendocrine tumors are often missed due to their deep mucosal origin with normal overlying mucosa. We first present the case of a 46-year-old woman with anemia and epigastric discomfort who was found to have an incidental submucosal mass in the duodenal bulb on esophagogastroduodenoscopy (EGD), which on endoscopic ultrasound (EUS) with a fine needle biopsy (FNB) showed a neuroendocrine tumor. Imaging with CT, however, failed to detect the presence of the mass in the duodenum. Furthermore, a DOTATATE scan showed only a nonspecific signal near the liver. The patient then underwent an EGD-guided, laparoscopic, robot-assisted transduodenal resection of the tumor, together with the removal of enlarged peritumoral lymph nodes. Pathology showed a well-differentiated neuroendocrine tumor of the duodenal bulb with metastasis to one lymph node, which was confirmed via immunohistochemistry staining. The second case is of a 51-year-old female who presented with occasional constipation and rectal pain and was found to have a rectal polypoid lesion on her colonoscopy, jumbo biopsies of which revealed a NET. An EUS done for staging and endoscopic mucosal resection (EMR) revealed a grade 1 well-differentiated NET on pathology, which was confirmed by immunohistochemistry staining. These cases stress the need for timely, definitive diagnosis and intervention. Here, we discuss the clinical features and investigations of neuroendocrine tumors for early diagnosis and management.

Discordance between in silico & in vitro analyses of ACE inhibitory & antioxidative peptides from mixed milk tryptic whey protein hydrolysate.

ACE inhibitory and antioxidative peptides identified by LCMS/MS, from mixed milk (Bubalus bubalis and Bos taurus) tryptic whey protein hydrolysate, were compared with the in silico predictions. α la and ß lg sequences, both from Bubalus bubalis and Bos taurus, were used for in silico study. SWISS-PROT and BIOPEP protein libraries were accessed for prediction of peptide generation. Study observed gaps in the prediction versus actual results, which remain unaddressed in the literature. Many peptides obtained in vitro, were not reflected in in silico predictions. Differences in identified peptides in separate libraries were observed too. In in silico prediction, peptides with known biological activities were also not reflected. Predictions, towards generation of bioactive peptides, based upon in silico release of proteins and amino acid sequences from different sources and thereupon validation in relation to actual results has often been reported in research literature. Given that computer aided simulation for prediction purposes is an effective research direction, regular updating of protein libraries and an effectual integration, for more precise results, is critical. The gaps addressed between these two techniques of research, have not found any address in literature. Inclusion of more flexibility with the variables, within the tools being used for prediction, and a hierarchy based database with search options for various peptides, will further enhance the scope and strength of research.

Production and characterisation of whey protein hydrolysate having antioxidant activity from cheese whey.

BACKGROUND: Cheese whey is a rich by-product in nutritional terms, possessing components with high biological value, excellent functional properties, and an inert flavour profile. In the present study, mozzarella cheese whey was ultra-filtrated to remove lactose and mineral. The retentate was hydrolysed with food-grade enzyme alcalase and the hydrolysis conditions (pH, temperature and time) were optimised for maximum antioxidant activity using response surface methodology. RESULTS: Whey protein hydrolysed for 8 h at pH 9 and 55 °C showed a maximum antioxidant activity of 1.18 ± 0.015 µmol Trolox mg(-1) protein. The antioxidant peptides were further enriched by ultra-filtration through a 3 kDa membrane. Seven peptides - ß-Lg f(123-131), ß-Lg f(122-131), ß-Lg f(124-131), ß-Lg f(123-134), ß-Lg f(122-131), ß-Lg f(96-100) and ß-Lg f(94-100) - were identified by LC-MS/MS in the 3 kDa permeate of the hydrolysate. The incorporation of whey protein hydrolysate (WPH) in lemon whey drink (5-10 g L(-1)) increased the antioxidant activity from 76% to 90% as compared to control. CONCLUSION: Hydrolysis of ultra-filtrated retentate of whey can be an energy- and cost-effective method for the direct production of WPH from whey compared to the industrial production of WPH from whey protein concentrate. This study suggests that WPH with good nutritional and biological properties can be effectively used in health-promoting foods as a biofunctional ingredient.

Process optimisation for preparation of caseinophosphopeptides from buffalo milk casein and their characterisation.

Caseinophosphopeptides (CPPs) are multifunctional bioactive peptides containing phosphorylated seryl residues in their sequence. In the present study, method for the production of CPPs from buffalo milk casein was optimised and characterised for their sequence, calcium solubilising and calcium binding activities. Response surface methodology was used to optimise the conditions for hydrolysis of buffalo casein by trypsin to obtain maximum yield of CPPs. The optimum hydrolysis conditions were as follows: hydrolysis pH 7.5, temperature 37 °C, hydrolysis time 7.0 h. Under these conditions, the experimental yield obtained was 10.04±0.24%, which is slightly lower than value predicted by the model. These CPPs were able to solubilise 1.03±0.08 mg la/mg CPPs in presence of excess phosphate and bind 0.935 mg of Ca/mg of CPPs. Eight phosphopeptides, i.e., α(s1)-CN f (37-58) 2P; α(s1)-CN f (37-58) 3P; α(s1)-CN f (35-58) 2P; α(s1)-CN f (35-58) 3P; α(s2)-CN f (2-21) 4P; α(s2)-CN f (138-149) 1P; ß-CN f (2-28) 4P and ß-CN f (33-48) 1P were identified by LC-MS/MS which contained motif for binding of divalent minerals. The sequences of these CPPs differed from that of derived from bovine casein.

College cafeteria snack food purchases become less healthy with each passing week of the semester.

OBJECTIVE: Snacks, stress and parties all contribute to the weight gain ­ the elusive 'Freshman 15' ­ that some college-goers unfortunately experience. The present study examines how a` la carte snack choice changes on a university campus during each progressing week of the academic calendar. DESIGN: How a` la carte snack choices change on a university campus with each progressing week of the academic calendar was examined. SETTING: The data were collected from three large cafeterias (or dining halls) on Cornell University's campus during four semesters (Fall 2006, Spring 2007, Fall 2007 and Spring 2008), for 18 weeks in each semester. SUBJECTS: After the a` la carte snack items were divided into healthy snacks and unhealthy snacks, the percentage share for each food category was calculated. RESULTS: Within each semester, the unhealthy snack food choices increased consistently by 0?4% per week (b50?00418, P,0?01). Furthermore, a sharp (8 %) increase occurred in the final two weeks of the semester. In contrast, healthy snack food choices decreased by almost 4% (b520?0408, P,0?01) in the final two weeks during the fall semester. CONCLUSIONS: These results demonstrate an increased demand for hedonic, or unhealthy, snack foods as the college semester progresses and in particular at the very end of the semester. To counter this tendency towards unhealthy snacking, cafeterias and stores should make extra effort to promote healthy alternatives during the later weeks of the semester.