Assessment of hemolytic activity, enzyme production and bacteriocin characterization of Bacillus subtilis LR1 isolated from the gastrointestinal tract of fish.

In the present investigation, probiotic potential (antagonistic activity, enzyme production, hemolytic activity, biosafety, antibiotic sensitivity and bile tolerance level) of Bacillus subtilis LR1 was evaluated. Bacteriocin produced by the bacterial strain B. subtilis LR1 isolated from the gastrointestinal tract of Labeo rohita was purified and characterized. The molecular weight of the purified bacteriocin was ~50 kDa in 12 % Native PAGE and showed inhibitory activity against four fish pathogens such as Bacillus mycoides, Aeromonas salmonicida, Pseudomonas fluorescens and Aeromonas hydrophila. The purified bacteriocin was maximally active at temperature 40 °C and pH 7.0, while none of the tested surfactants affect the bacteriocin activity. Extracellular enzyme activity of the selected bacterial strain was also evaluated. Amylase activity was estimated to be highest (38.23 ± 1.15 µg of maltose liberated mg-1 protein ml-1 of culture filtrate) followed by cellulase and protease activity. The selected bacterium was sensitive to most of the antibiotics used in this experiment, can tolerate 0.25 % bile salt and non-hemolytic in nature. Finally, the efficiency of the proposed probiotic candidate was evaluated in in vivo condition. It was detected that the bacterial strain can effectively reduce bacterial pathogenicity in Indian major carps.

Autochthonous Gut Bacteria in Two Indian Air-breathing Fish, Climbing Perch (Anabas testudineus) and Walking Catfish (Clarias batrachus): Mode of Association, Identification and Enzyme Producing Ability.

Scanning electron microscopy (SEM) was used to define the location of epithelium-associated bacteria in the gastrointestinal (GI) tract of two Indian air-breathing fish, the climbing perch (Anabas testudineus) and walking catfish (Clarias batrachus). The SEM examination revealed substantial numbers of rod shaped bacterial cells associated with the microvillus brush borders of enterocytes in proximal (PI) and distal regions (DI) of the GI tract of both the fish species. Ten (two each from the PI and DI of climbing perch and three each from the PI and DI of walking catfish) isolated bacterial strains were evaluated for extracellular protease, amylase and cellulase production quantitatively. All the bacterial strains exhibited high cellulolytic activity compared to amylolytic and proteolytic activites. Only two strains, CBH6 and CBH7, isolated from the DI of walking catfish exhibited high proteolytic activity. Maximum cellulase activity was exhibited by the strain, CBF2, isolated from the PI of climbing perch. Six most promising enzyme-producing adherent bacterial strains were identified by 16S rDNA gene sequence analysis. The strain ATH1 (isolated from climbing perch) showed high similarity fo Bacillus amyloliquefaciens whereas, the remaining five strains (isolated from walking catfish) were most closely related to Bacillus licheniformis.

Feasibility of a "No-Biopsy" Approach for the Diagnosis of Celiac Disease in Symptomatic Adults.

Celiac disease (CD) is an immune-mediated enteropathy, caused by hypersensitivity to gluten in genetically predisposed individuals. The worldwide prevalence of CD has been estimated to be approximately 1%. Most guidelines for diagnosis of CD rely on a sequential approach, with serological testing of antibodies against tissue transglutaminase (tTG) as a first-line test, followed by a duodenal biopsy. However, GI biopsy is an invasive procedure with various complications. Hence, this study was planned to ascertain whether it could be possible to have a non-biopsy approach, using only serological markers to establish the diagnosis of CD in adults. MATERIAL AND METHODS: It was a retrospective analysis of medical records of all biopsy-diagnosed CD patients with available anti-tTGA antibodies reports from 2019 to 2023. The patients were divided into three groups based on Marsh grading and anti-tTGA antibody levels were compared using various statistical tests. RESULTS: A total of 94 biopsy-diagnosed symptomatic CD patients with anti-tTGA antibody reports available formed the study group. Of these, 54 had biopsy findings consistent with Marsh 3 lesion, three had Marsh 2 lesion, and 37 had Marsh 1 lesion. A significant correlation existed between Marsh grading 3 lesion and anti-tTGA antibody levels above the upper limit of normal (ULN) x 10. CONCLUSION: Serum levels of anti-tTGA antibodies greater than 10 x ULN can be used to identify symptomatic patients with Marsh grade 3 CD lesions.

A kirigami-enabled electrochromic wearable variable-emittance device for energy-efficient adaptive personal thermoregulation.

For centuries, people have put effort to improve the thermal performance of clothing to adapt to varying temperatures. However, most clothing we wear today only offers a single-mode insulation. The adoption of active thermal management devices, such as resistive heaters, Peltier coolers, and water recirculation, is limited by their excessive energy consumption and form factor for long-term, continuous, and personalized thermal comfort. In this paper, we developed a wearable variable-emittance (WeaVE) device, enabling the tunable radiative heat transfer coefficient to fill the missing gap between thermoregulation energy efficiency and controllability. WeaVE is an electrically driven, kirigami-enabled electrochromic thin-film device that can effectively tune the midinfrared thermal radiation heat loss of the human body. The kirigami design provides stretchability and conformal deformation under various modes and exhibits excellent mechanical stability after 1,000 cycles. The electronic control enables programmable personalized thermoregulation. With less than 5.58 mJ/cm2 energy input per switching, WeaVE provides 4.9°C expansion of the thermal comfort zone, which is equivalent to a continuous power input of 33.9 W/m2. This nonvolatile characteristic substantially decreases the required energy while maintaining the on-demand controllability, thereby providing vast opportunities for the next generation of smart personal thermal managing fabrics and wearable technologies.

Protein extraction from Saccharomyces cerevisiae at different growth phases.

Saccharomyces cerevisiae is an established model organism with a well characterized genome. However, this model presents a unique problem due to a very resistant cell wall which develops in the late stationary phase resulting in sub-optimal extraction of proteins from such cells using majority of the cell lysis protocols. In this study, several methods from the literature with modifications thereof for lysis of S. cerevisiae cells were analyzed for their suitability for redox proteomics and biological activity studies of both exponential and late stationary phase cultures. The protocols applied are glass bead lysis, sonication, their combinations, alkali extraction, hot-SDS extraction methods and their modifications. The glass bead lysis method showed low yield but could be convenient in cases where in vitro processing steps post extraction is required or if only hydrophilic proteins are of interest. Hot-SDS and alkali extraction protocols yielded higher amount of proteins and these methods are potentially suitable for Western blotting and redox proteomic studies but allow no post-processing treatment(s) on the extracts which may be required for aging- and oxidative stress-related or other studies.

Integration of daytime radiative cooling and solar heating for year-round energy saving in buildings.

The heating and cooling energy consumption of buildings accounts for about 15% of national total energy consumption in the United States. In response to this challenge, many promising technologies with minimum carbon footprint have been proposed. However, most of the approaches are static and monofunctional, which can only reduce building energy consumption in certain conditions and climate zones. Here, we demonstrate a dual-mode device with electrostatically-controlled thermal contact conductance, which can achieve up to 71.6 W/m2 of cooling power density and up to 643.4 W/m2 of heating power density (over 93% of solar energy utilized) because of the suppression of thermal contact resistance and the engineering of surface morphology and optical property. Building energy simulation shows our dual-mode device, if widely deployed in the United States, can save 19.2% heating and cooling energy, which is 1.7 times higher than cooling-only and 2.2 times higher than heating-only approaches.

Role of Catalase in Oxidative Stress- and Age-Associated Degenerative Diseases.

Reactive species produced in the cell during normal cellular metabolism can chemically react with cellular biomolecules such as nucleic acids, proteins, and lipids, thereby causing their oxidative modifications leading to alterations in their compositions and potential damage to their cellular activities. Fortunately, cells have evolved several antioxidant defense mechanisms (as metabolites, vitamins, and enzymes) to neutralize or mitigate the harmful effect of reactive species and/or their byproducts. Any perturbation in the balance in the level of antioxidants and the reactive species results in a physiological condition called "oxidative stress." A catalase is one of the crucial antioxidant enzymes that mitigates oxidative stress to a considerable extent by destroying cellular hydrogen peroxide to produce water and oxygen. Deficiency or malfunction of catalase is postulated to be related to the pathogenesis of many age-associated degenerative diseases like diabetes mellitus, hypertension, anemia, vitiligo, Alzheimer's disease, Parkinson's disease, bipolar disorder, cancer, and schizophrenia. Therefore, efforts are being undertaken in many laboratories to explore its use as a potential drug for the treatment of such diseases. This paper describes the direct and indirect involvement of deficiency and/or modification of catalase in the pathogenesis of some important diseases such as diabetes mellitus, Alzheimer's disease, Parkinson's disease, vitiligo, and acatalasemia. Details on the efforts exploring the potential treatment of these diseases using a catalase as a protein therapeutic agent have also been described.

Evaluation of In Vivo Probiotic Efficiency of Bacillus amyloliquefaciens in Labeo rohita Challenged by Pathogenic Strain of Aeromonas hydrophila MTCC 1739.

Diseases in aquatic organisms, including fish, are a major concern in aquaculture production. In this present investigation, we have evaluated the beneficial effects of dietary Bacillus amyloliquefaciens CCF7 (GenBank Acc. No. KP256501) supplementation in rohu (Labeo rohita) challenged by a pathogenic strain of Aeromonas hydrophila MTCC 1739. Four experimental diets were formulated: control diet (no probiotics) and three experimental diets (different concentrations of probiotic candidate B. amyloliquefaciens CCF7 at 105 (T1), 107 (T2), 109 (T3) CFU/g). Further, we have divided the feeding trial into pre-challenge (70 days) and post-challenge (28 days) periods and various immune parameters (serum protein, globulin, albumin, lysozyme, and IgM), and stress parameters (malondialdehyde, catalase, and superoxide dismutase) were examined during both the periods. Throughout the entire experiment, control group was fed with probiotic free basal diet, while the treatment groups received probiotic supplemented diets (PSD). After challenge test, serum aspartate transaminase (AST), serum alanine transaminase (ALT) activity, and liver malondialdehyde level have increased significantly in control groups; however, level of these parameters were considerably lower in fish fed with PSD. In contrast, liver catalase and superoxide dismutase activities and serum globulin concentration was significantly higher in the group fed with T3 diet followed by T2. Furthermore, an elevated level of serum IgM and higher activity of serum lysozyme was also recorded in PSD fed groups, especially for T3 group which confirmed the probiotic efficiency of the bacterium B. amyloliquefaciens CCF7. We strongly believe that B. amyloliquefaciens CCF7 will be a good probiotic candidate in aquaculture industries.

Probiotic Potential of Autochthonous Bacteria Isolated from the Gastrointestinal Tract of Four Freshwater Teleosts.

In this study, a total of 121 bacterial strains were isolated from the gastrointestinal tract of four teleostean species, namely striped snakehead (Channa striatus), striped dwarf catfish (Mystus vittatus), orangefin labeo (Labeo calbasu) and mrigal carp (Cirrhinus mrigala), among which 8 isolates showed promising antibacterial activity against four potential fish pathogens, Aeromonas hydrophila, Aeromonas salmonicida, Aeromonas sobria and Pseudomonas fluorescens and were non-hemolytic. The isolates were further screened in response to fish bile tolerance and extracellular digestive enzyme activity. Two bacterial strains MVF1 and MVH7 showed highest tolerance and extracellular enzymes activities, and selected for further studies. Antagonistic activity of these two isolates was further confirmed by in vitro growth inhibition assay against four selected fish pathogens in liquid medium. Finally, these two bacterial strains MVF1 and MVH7 were selected as potential probiotic candidates and thus identification by partial 16S rRNA gene sequence analysis. The bacterial isolates MVF1 and MVH7 were identified as two strains of Bacillus sp.