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This study presents a genome analysis identifying various alkaline protease genes and culture condition optimization to enhance the production of the enzyme in Streptomyces barkulensis RC1831. The analysis of genes using bioinformatics tools such as RAST, KEGG, KAAS, and BLASTx revealed the presence of 10 genes that are responsible for alkaline protease production belonging to different families. Multiple sequence alignment showed a high sequence identity with M24 family serine protease followed by subtilisin-like serine protease. Optimal conditions for enhanced protease production were determined to be at 37°C, pH 11, casein1 % (W/V), dextrose 0.5 % (W/V), urea 0.5 % (W/V), tryptophan 1 % (W/V), 1 mM Mn+2, 1 % (V/V) Tween-80 in LB medium and an incubation time of 72 hours. Out of the 10 alkaline protease genes, 2 genes expressed significantly according to the activity observed during optimization processes i.e., M24 family, Mn+2 dependent metalloprotease and subtilisin-like serine, ca+2 dependent metalloprotease. Furthermore, the AP enzyme was remarkably stable in the presence of various cofactors (Metal ions) and surfactants, indicating its potential for industrial applications under diverse conditions.

Salinity and drought are the major abiotic stresses that significantly affect growth and productivity, necessitating the development and exploration of technologies to enhance tolerance of the crop plants. This study explored the role of four plant growth-promoting Streptomyces, i.e. Streptomyces chilikensis RC1830 (assigned as A), Streptomyces griseoincarnatus RB7AG (assigned as D), Streptomyces barkulensis RC1831 (assigned as B), and Streptomyces chitinovorans RC1832 (assigned as C) in mitigating stresses and improving rice plant growth (var. Swarna). Rice plants were inoculated with different Streptomyces consortia and exposed to 40 mM NaCl for salinity stress and 55 % FC (Field capacity) for drought stress. The study found that stresses adversely affected physiological parameters and triggered the production of antioxidant enzymes like superoxide dismutase, catalase, and peroxidase. However, inoculation of consortia at a concentration of 108 CFU/mL not only promoted the growth of the plants but also mitigated the negative effects of salinity and drought compared to non-inoculated controls. Among the treatments, ABCD consortia (RC1830, RC1831, RC1832, RB7AG) demonstrated the most significant improvements followed by ABD (RC1830, RC1831, RB7AG), boosting shoot length, root length, total chlorophyll, and proline content. This also enhanced superoxide dismutase, peroxidase, and catalase activities under stress conditions. Overall, Streptomyces inoculation improved stress tolerance in rice plants by enhancing photosynthetic capacity, antioxidant defense mechanisms, and proline content. These PGPS inoculants present a promising ecological approach to mitigate the impacts of abiotic stress on crops.

L-asparaginase (ASNase) obtained from microbial origins is a significant enzyme in the commercial sphere, crucial for the treatment of acute lymphoblastic leukemia. In the current investigation, we isolated ASNase through techniques involving ammonium sulfate precipitation and DEAE sepharose anion exchange column chromatography, revealing a molecular weight of 35 kDa. The enzyme exhibited its peak activity at pH 8 and 40°C, maintaining approximately 99% of its enzymatic effectiveness. The Michaelis constant (Km) and maximum velocity (Vmax) were determined to be 0.0021 M and 714.3 µM/min, respectively. ASNase was found to be non-toxic on HEK293 cells and selectively effective against breast cancer, hepatocellular carcinoma cells, colorectal adenocarcinoma cells, Human lung cancer cell line, Neck squamous cells, and Human head carcinoma cells. Hepatocellular carcinoma cells exhibited remarkable responsiveness to ASNase, displaying an IC50 of 23 µg/mL. This led to observable changes in nuclear morphology indicative of apoptosis, as evidenced by DAPI and Annexin V-FITC staining. ASNase also showed cell migration potential on HEK293 cells. Streptomyces HB2AG ASNase’s dynamic functioning across pH and temperature ranges, as well as its stability at 40°C and cytotoxic potential, make it a promising pharmaceutical candidate for various applications.

Introduction Atherosclerosis mediated by inflammatory markers is the corner stone in the pathology of coronary artery disease (CAD). Hyperlipidemia, one of the risk factors is treated with statins. Statins also have a pleotropic role in reducing inflammation. Effect of statins on two inflammatory markers pentraxin 3(PTX 3) and high sensitivity C-reactive protein (hs-CRP) is explored in this study.

Background Cervical cancer is a major public health problem in India, where access to prevention programmes is low. The WHO-Strategic Advisory Group of Experts recently updated their recommendation for human papillomavirus (HPV) vaccination to include a single-dose option in addition to the two-dose option, which could make HPV vaccination programmes easier to implement and more affordable.Methods We combined projections from a type-specific HPV transmission model and a cancer progression model to assess the health and economic effects of HPV vaccination at national and state level in India. The models used national and state-specific Indian demographic, epidemiological and cost data, and single-dose vaccine efficacy and immunogenicity data from the International Agency for Research on Cancer India vaccine trial with 10-year follow-up. We compared single-dose and two-dose HPV vaccination for a range of plausible scenarios regarding single-dose vaccine protection, coverage and catch-up. We used a healthcare sector payer perspective with a time horizon of 100 years.Results Under the base-case scenario of lifelong protection of single-dose vaccination in 10-year-old girls with 90% coverage, the discounted incremental cost-effectiveness ratio (ICER) of nationwide vaccination relative to no vaccination was US$406 (₹INR30 000) per DALY (disability-adjusted life-years) averted. This lay below an opportunity-cost-based threshold of 30% Indian gross domestic product per capita in each Indian state (state-specific ICER range: US$67–US$593 per DALY averted). The ICER of two-dose vaccination versus no vaccination vaccination was US$1404 (₹INR104 000). The ICER of two-dose vaccination versus single-dose vaccination, assuming lower initial efficacy and waning of single-dose vaccination, was at least US$2282 (₹INR169 000) per DALY averted.Conclusions Nationwide introduction of single-dose HPV vaccination at age 10 in India is highly likely to be cost-effective whereas extending the number of doses from one to two would have a less favourable profile.

Abstract Background Nowadays, the conventional plastic wastes are very challenging to environments and its production cost also creates an economic crisis due to petrochemical-based plastic. In order to solve this problem, the current studies were aimed at screening and characterizing these polyhydroxyalkanoate (PHA)-producing isolates and evaluating the suitability of some carbon source for newly screened PHA-producing isolates. Material and methods Some carbon sources such as D-fructose, glucose, molasses, D-ribose and sucrose were evaluated for PHA production. Data were analyzed using SPSS version 20. The 16SrRNA gene sequence of these isolates was performed. These newly isolated taxa were related to Bacillus species. It was designated as Bacillus sp. LPPI-18 and affiliated Bacillus cereus ATCC 14577T (AE01687) (99.10%). Paenibacillus sp. 172 (AF273740.1) was used as an outgroup. Results Bacillus sp. LPPI-18 is a gram-positive, rod-shaped, endospore former, and citrate test positive. This isolate showed positive for amylase, catalase, pectinase, and protease test. They produced intracellular PHA granules when this isolate was stained with Sudan Black B (SBB) and Nile blue A (NBA) preliminary and specific staining dyes, respectively. Both temperature and pH used to affect polyhydroxyalkanoates (PHA) productivity. Bacteria are able to reserve PHA in the form of granules during stress conditions. This isolate produces only when supplied with carbon sources. More PHA contents (PCs) were obtained from glucose, molasses, and D-fructose. In this regard, the maximum mean value of PC was obtained from glucose (40.55±0.7%) and the minimum was obtained from D-ribose (12.4±1.4%). Great variations (P≤0.05) of PCs were observed among glucose and sucrose, molasses and sucrose, and D-fructose and sucrose carbon sources for PHA productivity (PP) of cell dry weight (CDW) g/L. After extraction, PHA film was produced for this typical isolate using glucose as a sole carbon source. Fourier transform infrared spectrum was performed for this isolate and showed the feature of polyester at 1719.64 to 1721.16 wavelengths for these extracted samples. The peak of fingerprinting (band of carboxylic acid group) at this wavelength is a characteristic feature of polyhydroxybutyrate (PHB) and corresponds to the ester functional group (C=O). Conclusion In this study, newly identified Bacillus sp. LPPI-18 is found to be producing biodegradable polymers that are used to replace highly pollutant conventional plastic polymers. This isolate is also used to employ certain cost-effective carbon sources for the production of PHA polymers. Graphical Abstract

Background: Psoriasis has well-known associations with individual components of metabolic syndrome such as hypertension, dyslipidemia, insulin resistance, and obesity. Traditional anthropometric measurements of obesity such as waist circumference (WC) and body mass index (BMI) do not differentiate between subcutaneous fat and visceral fat, the latter being associated with cardiovascular risk factors. Lipid accumulation product (LAP) index is a measure of visceral fat and has been found to be a better predictor of cardiovascular risk. However, LAP index has not been well-studied in psoriasis patients. Aims and Objectives: Our objective was to find out if LAP index differs significantly between psoriasis patients and controls and whether LAP index shows a correlation with duration and severity of psoriasis. Materials and Methods: A case–control study was undertaken with 40 chronic plaque psoriasis patients and 42 controls. BMI and LAP index were calculated for all the patients and controls. Psoriasis area severity index (PASI) was calculated for all the psoriasis patients. Mann–Whitney U-test was done for comparing the age, BMI, WC, serum triglyceride, and LAP index between the cases and controls and to compare the LAP index between mild psoriasis and moderate-to-severe psoriasis groups. Spearman's correlation coefficient was used to assess the correlation of LAP index with duration of psoriasis and with PASI. Logistic regression models were done to assess the risk factors in psoriasis. Results: A statistically significant difference was observed between the LAP index of controls (23.79 ± 13.02) and that of psoriasis patients (46.42 ± 27.2). LAP index was significantly higher in the moderate-to-severe psoriasis group as compared to the mild psoriasis group. LAP index was a significant risk factor associated with psoriasis (OR = 1.07; 95% CI: 1.03 – 1.11). Conclusion: Calculation of LAP index in psoriasis patients helps in identification of more individuals at high risk of cardiovascular morbidity than traditional anthropometric measurements of obesity.

Abstract Background WHO, UNODC, and UNAIDS recommend a comprehensive package for prevention, treatment, and care of HIV among people who inject drugs (PWID). We describe the uptake of services and the cost of implementing a comprehensive package for HIV prevention, treatment, and care services in Delhi, India. Methods A cohort of 3774 PWID were enrolled for a prospective HIV incidence study and provided the comprehensive package: HIV and hepatitis testing and counseling, hepatitis B (HB) vaccination, syndromic management of sexually transmitted infections, clean needles-syringes, condoms, abscess care, and education. Supplementary services comprising tea and snacks, bathing facilities, and medical consultations were also provided. PWID were referred to government services for antiretroviral therapy (ART), TB care, opioid substitution therapy, and drug dependence treatment/rehabilitation. Results The project spent USD 1,067,629.88 over 36 months of project implementation: 1.7% on capital costs, 3.9% on participant recruitment, 26.7% for project management, 49.9% on provision of services, and 17.8% on supplementary services. Provision of HIV prevention and care services cost the project USD 140.41/PWID/year. 95.3% PWID were tested for HIV. Of the HIV-positive clients, only 17.8% registered for ART services after repeated follow-up. Reasons for not seeking ART services included not feeling sick, need for multiple visits to the clinic, and long waiting times. 61.8% of the PWID underwent HB testing. Of the 2106 PWID eligible for HB vaccination, 81% initiated the vaccination schedule, but only 29% completed all three doses, despite intensive follow-up by outreach workers. PWID took an average of 8 clean needles-syringes/PWID/year over the project duration, with a mid-project high of 16 needles-syringes/PWID/year. PWID continued to also procure needles from other sources, such as chemists. One hundred five PWID were referred to OST services and 267 for rehabilitation services. Conclusions A comprehensive HIV prevention, treatment, and care package is challenging to implement. Extensive efforts are needed to ensure the uptake of and retention in services for PWID; peer educators and outreach workers are required on a continuous basis. Services need to be tailored to client needs, considering clinic timing and distance from hotspots. Programs may consider provision of ART services at selected drop-in centers to increase uptake.

An extracellular thermo-alkali stable chitinase was obtained from Streptomyces chilikensis RC1830, a novel actinobacterial strain isolated from the sediments of Chilika lake, India. Purification of the enzyme was carried out by concentrating the enzyme with centrifugal device followed by chromatographic separation by DEAE Sepharose ion exchange resin.The molecular weight of the enzyme was 10.5 kDa as determined by SDS-PAGE. The optimum pH and temperature for the partially purified chitinase was pH 7 and 60 °C. The chitinase showed 40% activity at pH 11 after 24 h exposure at room temperature. The chitinase exhibited Km and Vmax values are 0.02 mM and 3.184 mol/min/mg of enzyme respectively. The 6 residue N-terminal sequence of the enzyme was not found similar to any of the reported chitinase enzyme. Based on the SDS PAGE, zymogram analysis, activity assays and other characteristics, it is proposed that the purified enzyme from S.chilikensis RC1830 is a chitinase. Keywords: Streptomyces, Chitinase, Halo tolerant, Chio oligosaccharides

Background: A controversy exists regarding the association between subclinical hypothyroidism (SH) and dyslipidemia. Moreover, studies on lipid ratios and atherogenic index of plasma (AIP) in SH are rare, particularly in the Indian scenario. Aim: This study aimed to investigate abnormalities in conventional lipid profile, lipid ratios, and AIP in SH and attempted to correlate thyroid stimulating hormone (TSH) and AIP in SH. Materials and Methods: In this retrospective analysis of patient records of SH subjects and euthyroid subjects, age, free triiodothyronine, free thyroxine, TSH, total cholesterol, triglycerides, high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol, lipid ratios, and AIP were compared between the two groups. The correlation of TSH and AIP in SH was studied. Spearman′s correlation, Mann-Whitney U-test and logistic regression analysis were performed. Results: Triglyceride, triglyceride/HDL-C, and AIP were significantly higher in SH as compared to euthyroid group, but there was no correlation between TSH and AIP in SH. AIP emerged as the significant single factor associated with SH in multiple logistic regressions. Conclusion: The positive association of dyslipidemia and SH indicates a need for regular screening of these patients to enable early diagnosis and treatment of dyslipidemia. Even in patients who have a normal conventional lipid profile, lipid ratios, and AIP have to be calculated for better assessment of atherogenic risk.

Factitious test reports may result in incorrect diagnosis and incorrect management. Such incorrect diagnosis can be prevented by a vigilant biochemist. We report a case of Rhabdomyolysis presenting with extremely low total Creatine Kinase (CK) levels which was factitious. Running the sample in dilution resulted in a very high value of total CK which could have been missed if the sample was not run in dilution and the diagnosis of Rhabdomyolysis could have been missed.

Introduction: Previous studies indicate a higher prevalence of metabolic syndrome in psoriatic patients. This study aimed to investigate the association of metabolic syndrome and its components with psoriasis. It also studied the relation between presence of metabolic syndrome and disease duration in psoriasis patients. Materials and Methods: This was a hospital-based, casecontrol study conducted with 30 clinically diagnosed patients of chronic plaque psoriasis and 30 age- and sex-matched control subjects. Height, weight, BMI, blood pressure and waist circumference were assessed in all the subjects. Fasting levels of serum glucose, serum triglycerides and serum HDL were estimated by automated clinical chemistry analyser. Metabolic syndrome was diagnosed by the presence of at least 3 criteria of NCEP ATP III with Asian modification for waist circumference. Results: Metabolic syndrome was more common in psoriatic patients than in controls but the difference was statistically insignificant (60% vs. 40%, p-value=0.12). The psoriasis group had a higher prevalence of elevated blood glucose levels and higher waist circumference compared to controls. Psoriasis patients had a higher prevalence of high triglyceride levels than controls, the difference being statistically insignificant (40% vs. 30%, p-value = 0.41). The prevalence of low HDL levels was significantly higher in cases compared to controls (86.7% vs. 60%, p-value = 0.02). There was no relation between presence of metabolic syndrome and duration of psoriasis. Conclusion: Our findings suggest that metabolic syndrome as well as dyslipidaemia is commoner in psoriasis patients. This underlines the need for screening of all psoriasis patients for early diagnosis and treatment of associated metabolic syndrome to reduce the high burden of morbidity and mortality.

Introduction: Laboratory errors are result of a poorly designed quality system in the laboratory. Six Sigma is an error reduction methodology that has been successfully applied at Motorola and General Electric. Sigma (σ) is the mathematical symbol for standard deviation (SD). Sigma methodology can be applied wherever an outcome of a process has to be measured. A poor outcome is counted as an error or defect. This is quantified as defects per million (DPM). A six sigma process is one in which 99.999666% of the products manufactured are statistically expected to be free of defects. Six sigma concentrates, on regulating a process to 6 SDs, represents 3.4 DPM (defects per million) opportunities. It can be inferred that as sigma increases, the consistency and steadiness of the test improves, thereby reducing the operating costs. We aimed to gauge performance of our laboratory parameters by sigma metrics. Objectives: Evaluation of sigma metrics in interpretation of parameter performance in clinical biochemistry. Material and Methods: The six month internal QC (October 2012 to march 2013) and EQAS (external quality assurance scheme) were extracted for the parameters-Glucose, Urea, Creatinine, Total Bilirubin, Total Protein, Albumin, Uric acid, Total Cholesterol, Triglycerides, Chloride, SGOT, SGPT and ALP. Coefficient of variance (CV) were calculated from internal QC for these parameters. Percentage bias for these parameters was calculated from the EQAS. Total allowable errors were followed as per Clinical Laboratory Improvement Amendments (CLIA) guidelines. Sigma metrics were calculated from CV, percentage bias and total allowable error for the above mentioned parameters. Results: For parameters - Total bilirubin, uric acid, SGOT, SGPT and ALP, the sigma values were found to be more than 6. For parameters – glucose, Creatinine, triglycerides, urea, the sigma values were found to be between 3 to 6. For parameters – total protein, albumin, cholesterol and chloride, the sigma values were found to be less than 3. Conclusion: ALP was the best performer when it was gauzed on the sigma scale, with a sigma metrics value of 8.4 and chloride had the least sigma metrics value of 1.4.

Introduction: Electrolyte abnormalities are one of the common causes of morbidity and mortality in critically ill patients. The turnaround time for electrolyte reporting should be as low as possible. Electrolytes are measured conventionally in serum obtained from venous blood by electrolyte analyser which takes 20 to 30 min. Point of care analysers are now available where in electrolytes can be measured in arterial blood within 5 min. This study was done to study the agreement of arterial sodium and arterial potassium with venous sodium and venous potassium levels. Materials and Methods: Venous sodium and venous potassium levels and arterial sodium and arterial potassium levels were analysed on 206 patient samples admitted to Intensive Care Unit (ICU). The venous values were compared with the arterial values for correlation. Venous sodium was compared with arterial sodium by spearman correlation. Venous potassium was compared with arterial potassium by pearson correlation. Results: The mean value of arterial sodium was 134 and venous sodium was 137. The mean value of arterial potassium was 3.6 and venous potassium was 4.1. The correlation coefficient obtained for sodium was 0.787 and correlation coefficient obtained for potassium was 0.701. There was positive correlation of arterial sodium and arterial potassium with venous sodium and venous potassium indicating agreement between the parameters. Conclusion: Arterial sodium and arterial potassium can be used instead of venous sodium and venous potassium levels in management of critically ill patients.

ABSTRACT Factitious biochemical reports result in the misguiding of clinicians, unnecessary retesting, wrong diagnoses and incorrect treatments. A vigilant biochemist identifies these factitious biochemical reports and alerts the clinician regarding the proper interpretation of the biochemical reports, thus preventing a misdiagnosis and an incorrect treatment. We are presenting a case report of a multiple myeloma patient who presented with factitious biochemical reports which were caused due to paraproteinaemia. In the present case, the patient presented with an underestimation of urea and creatinine, an underestimation of sodium, low albumin levels and high phosphate levels. On repeating the same tests after dilutions and deproteinizing, the effects of the paraproteins on the above mentioned tests were reduced. Thus, from the observations of our study, we suggest that the interference by paraproteinaemia can be reduced by analyzing the biochemical parameters after dilution and deproteinization.

Introduction: The introduction of a new method or new analyser is a common occurrence in clinical biochemistry laboratory. Blood gas measurements and electrolytes are often performed in Point-of-Care (POC) settings. When a new POC analyser is obtained, the performance of the analyser should be evaluated by comparison to the measurements with the reference analyser in the laboratory. Objectives: Evaluation of method performance of pH, PCO2 , PO2 , Na+, K+ of cobas b121 ABG analyser. Materials & Methods: The evaluation of method performance of pH, PO2 , PCO2 , Na+, K+ of cobas b121 ABG analyser was done by comparing the results of 50 patient samples run on cobas b121 with the results obtained from Rapid lab values (reference analyser). Correlation coefficient was calculated from the results obtained from both the analysers. Precision was calculated by running biorad ABG control samples. Results: The correlation coefficient values obtained for parameters were close to 1.0 indicating good correlation. The CV obtained for all the parameters were less than 5 indicating good precision. Conclusion: The new ABG analyser, Cobas b121 correlated well with the reference ABG analyser (Rapid Lab) and could be used to run on patient samples.

People who inject drugs (PWIDs) are at risk of exposure to blood borne pathogens. India has a large PWID population and the highest HIV transmission is among PWIDs. Interventions to prevent risky injection is constantly tested by the scientific community and innovations are required in this area. 3748 male PWIDs were recruited into a longitudinal study. 2509 PWIDs who participated in the end of study survey, also responded to multiple open ended questions asked by the interviewers. The qualitative interviews were analyzed focusing on identifying themes and relationship among themes. 35.6% of the cohort had engaged in risky injection at least once. Although the project activities promoted safe injection practices among majority of the study participants; 35% is a substantial population at risk. Gaining knowledge about blood borne pathogen transmission, motivated participants to be careful about not sharing injection or paraphernalia. The belief that friends are safe made many share among themselves. Not prepared when the withdrawal set in was the main reason for sharing injection. The project intervention had stimulating effect on the PWIDs and they tried to stop injection and follow safe injection practices. However, a failure at least once is an exposure to blood borne pathogens. Hence teaching PWIDs to plan for their injection practice and keeping materials ready at hand is important. [ABSTRACT FROM AUTHOR]