Estimation of extracellular lipase enzyme produced by thermophilic bacillus sp. isolated from arid and semi-arid region of Rajasthan, India

Thermophilic organisms can be defined as microorganisms which are adapted to live at high temperatures. The enzymes produce by thermophilic bacteria are capable of catalyzing biochemical reactions at high temperatures. Thermophilic bacteria are able to produce thermostable lipase enzymes capable of degradation of lipid at temperatures higher than those of mesophilic bacteria. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are quite useful in terms of discovering thermophilic lipase enzymes. Due to great temperature fluctuation in hot arid and semi-arid regions of Rajasthan, this area could serve as a good source for new thermophilic lipase producing bacteria with novel industrially important properties. The main objective of this research is the isolation and estimation of industrially important thermophilic lipase enzyme produced by thermophilic bacteria, isolated from arid and semi-arid region of Rajasthan. For this research purpose, soil samples were collected from Churu, Sikar and Jhunjunu regions of Rajasthan. A total of 16 bacterial strains were isolated, and among these bacterial isolates only two thermophilic lipase producing bacteria were identified. The thermophilic lipase enzyme was estimated by qualitative and quantitative experiments. The isolate was identified as Bacillus sp. by microscopic, biochemical and molecular characterization. The optimum enzyme activity was observed at pH 8, temperature 60°C and 5% salt concentration at 24 hrs time duration. Lipases are useful in a variety of biotechnological fields such as food and dairy (cheese ripening, flavour development), detergent, pharmaceutical (naproxen, ibuprofen), agrochemical (insecticide, pesticide) and oleochemical (fat and oil hydrolysis, biosurfactant synthesis) industries. Lipases can be further used in many newer areas where they can serve as potential biocatalysts. 
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Structural Modeling and Validation of Rep protein of Begomovirus Strains (TLCBV and TYLCTHV)

Homology modeling involves taking a known sequence with an unknown structure and mapping it against a known structure of one or several similar (homologous) proteins. It would be expected that two proteins of similar origin and function would have reasonable structural similarity. Therefore it is possible to use the known structure as a template for modeling the structure of the unknown structure. Proteins that share the same function generally have similar structures. During alignment if two proteins show maximum sequence identity they also show a similar folding pattern. This principal has became the foundation of homology modeling. 
The Geminivirus taxonomic group of plant viruses is characterized by geminate particles and genomes consisting of single-stranded circular DNA molecules of about 2.5 to 2.8 kb in size. Agricultural plants are threatened by many diseases caused by whitefly-transmitted geminiviruses. Since these diseases are in a fast spreading phase, it is urgent to devise rapid diagnosis methods and to produce resistant plants. 
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Antibacterial Mode of Action of the Essential Oil Obtained from Chamaecyparis obtusa Sawdust on the Membrane Integrity of Selected Foodborne Pathogens

U ovom je radu ispitan mehanizam antibakterijskog učinka esencijalnog ulja dobivenog iz piljevine pačempresa (Chamaecyparis obtusa) na patogene bakterije u hrani. Esencijalno je ulje dobiveno mikrovalnom ekstrakcijom i hidrodestilacijom piljevine pačempresa. Vrijednosti su minimalne inhibitorne koncentracije esencijalnog ulja što su sprečavale rast bakterija u hrani, poput Bacillus cereus ATCC 13061, Listeria monocytogenes ATCC 7644, Staphylococcus aureus ATCC 12600, Salmonella Typhimurium ATCC 43174 i Escherichia coli ATCC 43889, bile u rasponu od 62,5 do 500 μg/mL, a vrijednosti minimalne baktericidne koncentracije u rasponu od 125 do 1000 μg/mL. Potvrđeno je da je esencijalno ulje u minimalnoj inhibitornoj koncentraciji sprečavalo rast stanica ispitanih bakerija. Osim toga, pretražnom su elektronskom mikroskopijom pronađene bitne morfološke promjene ili puknuća stanične membrane bakterija B. cereus ATCC 13061 i E. coli ATCC 43889, čime je potvrđen inhibicijski utjecaj esencijalnog ulja iz piljevine pačempresa. Oslobađanje velikih količina izvanstaničnog adenozin 5’-trifosfata (ATP) i materijala koji se apsorbira na valnoj duljini od 260 nm, te gubitak iona kalija iz stanice negativno djeluju na Gram-pozitivnu bakteriju B. cereus ATCC 13061 i Gram-negativnu bakteriju E. coli ATCC 43889, što potvrđuje učinak esencijalnog ulja na staničnu membranu. Iz dobivenih se rezultata može zaključiti da esencijalno ulje dobiveno iz piljevine pačempresa ima antibakterijsku aktivnost širokog spektra, te da djeluje na integritet stanične membrane i morfološka svojstva bakterija izoliranih iz hrane.The present study examines the possible antibacterial mechanism of action of the essential oil obtained from Chamaecyparis obtusa (COEO) sawdust against foodborne pathogenic bacteria. The COEO was obtained by microwave-assisted hydrodistillation of C. obtusa sawdust. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of COEO against the tested foodborne pathogens including Bacillus cereus ATCC 13061, Listeria monocytogenes ATCC 7644, Staphylococcus aureus ATCC 12600, Salmonella Typhimurium ATCC 43174 and Escherichia coli ATCC 43889 were found in the range from 62.5 to 500 μg/mL and from 125 to 1000 μg/mL, respectively. At the MIC concentrations, the COEO had potential inhibitory effect on the cell viability of the tested bacteria. In addition, the scanning electron microscopic analysis confirmed the inhibitory effect of COEO by revealing significant morphological alterations or rupture of the cell membranes of B. cereus ATCC 13061 and E. coli ATCC 43889. Moreover, the mode of action of COEO on the cell membrane of both Gram-positive B. cereus ATCC 13061 and Gram-negative E. coli ATCC 43889 bacteria was confirmed by marked release of extracellular adenosine 5’-triphosphate (ATP) and cellular material that absorbs at 260 nm, and by efflux of potassium ions. These findings suggest that COEO holds a broad-spectrum antibacterial efficacy, confirming its influence on the membrane integrity and morphological characteristics of tested foodborne pathogens

A diterpenoid sugiol from Metasequoia glyptostroboides with α-glucosidase and tyrosinase inhibitory potential

Nowadays use of plant derived natural compounds have become a topic of increasing interest in food and medicine industries due to their multitude of biological and therapeutic properties. In this study, a diterpenoid compound sugiol, isolated from Metasequoia glyptostroboideswas evaluated for α-glucosidase and tyrosinase inhibitory efficacy in terms of its potent anti-diabetic and anti-melanogenesis potential, respectively. As a result, sugiol at the concentration range of (100-10,000 µg/mL) and (20-500 µg/mL) showed potent efficacy on inhibiting α-glucosidase and tyrosinase enzymes in vitro ranging from 12.34-63.47% and 28.22-67.43%, respectively. These findings confirm the therapeutic potential of diterpenoid compound sugiol from M. glyptostroboides as a novel candidate for using in food and medicine industry which may have practical potential to cure skin and diabetes mellitus type-2 related disorders

SplitEE: Early Exit in Deep Neural Networks with Split Computing

Deep Neural Networks (DNNs) have drawn attention because of their outstanding performance on various tasks. However, deploying full-fledged DNNs in resource-constrained devices (edge, mobile, IoT) is difficult due to their large size. To overcome the issue, various approaches are considered, like offloading part of the computation to the cloud for final inference (split computing) or performing the inference at an intermediary layer without passing through all layers (early exits). In this work, we propose combining both approaches by using early exits in split computing. In our approach, we decide up to what depth of DNNs computation to perform on the device (splitting layer) and whether a sample can exit from this layer or need to be offloaded. The decisions are based on a weighted combination of accuracy, computational, and communication costs. We develop an algorithm named SplitEE to learn an optimal policy. Since pre-trained DNNs are often deployed in new domains where the ground truths may be unavailable and samples arrive in a streaming fashion, SplitEE works in an online and unsupervised setup. We extensively perform experiments on five different datasets. SplitEE achieves a significant cost reduction ($>50\%$) with a slight drop in accuracy ($<2\%$) as compared to the case when all samples are inferred at the final layer. The anonymized source code is available at \url{https://anonymous.4open.science/r/SplitEE_M-B989/README.md}.Comment: 10 pages, to appear in the proceeding AIMLSystems 202

Acute on chronic liver failure- etiology, clinical profile, prognostic scores: experience from tertiary care centre of eastern India

Background: Acute on chronic liver failure (ACLF) is a recently recognised entity in chronic liver disease patients. Data regarding ACLF in terms of clinical presentation, etiology of underlying cirrhosis, precipitating factors, prognostic factors are lacking from eastern India. The present study was undertaken to evaluate the above factors along with the assessment of short-term mortality (4 weeks) in patients of ACLF.Methods: In this prospective observational study, 120 patients diagnosed as ACLF were included. A comparison of complications, biochemical profiles and prognostic scores was made between the survivor and non-survivor groups.Results: Of the 120 ACLF patients included, the mean age was 44.9±8.6 years and the male to female ratio was 2.5:1. Common clinical presentations were jaundice (100%), ascites (88.3%), hepatic encephalopathy (60%). The most common etiology for underlying CLD was alcohol (51.7%) followed by chronic hepatitis B (20%) and chronic hepatitis C (15%) infection. Alcohol hepatitis (40%) followed by hepatotropic viral infections (20%) and drug-induced liver injury (15%) were common identifiable precipitating agents. After a follow-up period of 4 weeks, 56 (46.6%) out of 120 patients died. The presence of sepsis, hyponatremia, renal failure, and coagulopathy was significantly associated with high mortality. Mortality was higher among patients having high Chronic liver failure consortium- acute on chronic liver failure (CLIF-ACLF) grade and closely related to the number of organ failures.Conclusions: ACLF is a rapidly progressive syndrome in chronic liver disease patients, having high short-term mortality

Isolation and preliminary characterization of a bacteriocin-producer Bacillus strain inhibiting methicillin resistant Staphylococcus aureus

In a multivalent approach to discover new antimicrobial substance, a total of 160 Bacilli were isolated from termitarium soil, characterized on the basis of their morphological and physiological characters and screened for their antimicrobial activity by agar well diffusion method against certain drug resistant pathogenic bacteria such as Staphylococcus aureus, Methicillin resistant Staphylococcus aureus and common food contaminating bacteria Listeria monocytogenes. After preliminary screening, sixteen isolates showed inhibitory activity against test pathogens. Among them Bacillus isolate TSH58 exhibited maximum inhibitory activity against MRSA, Staphylococcus aureus and Listeria monocytogenes. Based on morphological, physiological, biochemical and 16S rDNA characteristics isolate TSH58 was identified as a member of the Bacillus cereus species group. Various nutrient sources and culture conditions were optimized, the partially purified antimicrobial metabolite was subjected to various treatments such as heat, pH and proteolytic enzymes. Complete loss in the activity observed when the crude metabolite was treated with proteolytic enzymes suggesting its proteinaceous nature and termed as bacteriocin like inhibitory substance (BLIS). Minimal inhibitory concentration of the partially purified bacteriocin determined by microtiter plate assay was 80 μg/ml for MRSA and 40 μg/ml for L. monocytogenes. Tricine SDS PAGE analysis revealed that the partially purified bacteriocin produced by the Bacillus strain TSH58 had an apparent molecular weight of about 4.0 KDa

Experimental strategy of animal trial for the approval of anti-diabetic agents prior to their use in pre-human clinical trials

Although several naturally available drugs have been historically used for the treatment of diabetes mellitus throughout the world, few of them have been validated by scientific criteria. Before approval of any drug developed it should pass through animal trial prior to clinical human trial, which should followed by some standard ethical rules. Recently, a large diversity of animal models have been developed to better understand the pathogenesis of diabetes mellitus, and new drugs have been introduced in the market to treat this autoimmune disease. In the present article, we demonstrated some standard handling procedure of animal trial for the approval of anti-diabetic drug, which could be helpful for both academics and industrial scientific community to conduct the animal experiments. This research also contributes in the field of ethnopharmacology to design new strategies for the development of novel drugs to treat this serious condition of diabetes mellitus that constitutes a global public health. Video Clip of Methodology:  Handling and caring of mice: 2 min 30 sec   Full Screen   Alternate Inducing diabetes in mice and observing blood glucose level: 1 min 47 sec   Full Screen   Alternate Drug administration and observation of blood glucose level: 2 min 11 sec   Full Screen   Alternat

Imiquimod-induced psoriasis-like skin inflammation in mouse model

Psoriasis is a skin disorder characterized by skin inflammation and plaques. Induction of psoriasis in animal model include following steps: a) Selection of animal model, b) Hair removing from the back or ear, c) treatment of skin with Aldara, a cream containing 5% imiquimod and d) Observation. Imiquimod-induced skin inflammation in animal model resembles with psoriasis. Video Clip of Methodology: Psoriaisis-like skin: 9 min 10 sec   Full Screen   Alternat