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In this work, a psychrotrophic bacteria producing cold-active protease, was obtained from Dachigam National Park, an ecologically significant habitat in Western Himalayas owing to its varied endemic and endangered flora and fauna. This isolate was identified as Bacillus sp. HM49 via phenotypic, Gram staining, bio-chemical and 16S rRNA gene identification. Isolate HM49 when tested for proteolytic activity revealed prominent hydrolytic zone with the most production at 20 °C and pH, 8.0 post 72 h incubation. This enzyme was purified, enhancing its specific activity to 61.15 U/mg and its characterization studies revealed it to be a cold-alkaline protease being active in a wide pH (6.0-12) and temperature (5-40 °C) range. Amplification of CAASPR gene of HM49 was performed, followed by enzyme-substrate docking studies and MMGBSA providing details about its type, molecular weight validation as well as functional applications. The purified protease of HM49 was tested for laundry applications and the enzyme was found to be compatible with majority of the detergents tested. Its potential as an eco-friendly detergent additive was further validated by wash performance test as it effectively removed recalcitrant blood stains at a low temperature of 20 °C that could be beneficial for fine garments like silk which preferably need cold washing.

This paper examines the financial and social efficiency of the microcredit programs offered by the Pakistan Poverty Alleviation Fund partner organizations. Panel data concerning variables of interest are collected from Pakistan Microfinance Network, covering a minimum of 14 partner organizations (in 2005) to a maximum of 35 partner organizations (in 2014). The data is analyzed using the Data Envelopment Analysis, assuming both constant and variable returns to scale scenarios and the operational scale of the partner organizations. Trends in average efficiency scores have been analyzed to assess the mission drift of the partner organizations. Results reveal that managerial inefficiency is more pronounced than the sub-optimal production scale in all three scenarios under consideration. Moreover, trends in the efficiency scores indicated a slight mission drift of the microfinance providers. About 77.5% of the partner organizations were financially sustainable over the entire study period. The study recommends providing objective-oriented training, workshops, and seminars for managing microfinance providers.

BackgroundCoronavirus disease-19 (COVID-19) has not only spawned a lot of stigma and discrimination towards its survivors but also to their corpses. We aimed to assess the magnitude and correlates of stigma in these survivors, on return to their communities.MethodsThis was a cross-sectional, hospital-based, exploratory study conducted by the postgraduate department of psychiatry, in collaboration with the postgraduate department of chest medicine, Govt. medical college, Srinagar. The study was performed among COVID-19 survivors, who attended the outpatient department after their discharge from the hospital. Socio-demographic characteristics were recorded through semi-structured proforma. Stigma was measured by the stigma questionnaire. Data was analyzed using descriptive statistics and regression analysis.ResultsA total of 91 survivors consented to participate in the study. Almost half (46.2%) of them were in the age group of 30-49 years and close to two-thirds (68.1%) were males. About three-fourths (74.7%) were from the urban background. The mean time from hospital discharge to study entry was 11.7±5.1 [Range(R) = 7-21] days. 98% of survivors provided at least one stigma endorsing response and the total mean stigma score was 28.5±7.1[R = 6-39]. The mean stigma sub-scores were highest for enacted stigma (7.6±1.8) [R = 2-9] and externalized stigma (15.0±4.1) [R = 1-20]. Enacted stigma was significantly high in males as compared to females. Enacted stigma and internalized stigma were both associated with education. Enacted stigma, externalized stigma, disclosure concerns, and total stigma was significantly associated with the occupation. Being unemployed and time since discharge were identified as independent predictors of total stigma.ConclusionOur study results showed high levels of enacted and externalized stigma among COVID-19 survivors. Enacted stigma was more among males and in those who were highly educated. Survivor centered and community-driven anti-stigma programs are the need of the hour to promote the recovery and community re-integration of these survivors.

[This corrects the article DOI: 10.1371/journal.pone.0240152.].

Ralstonia solanacearum is a soil-borne plant pathogen that infects more than 200 plant species. Its broad host range and long-term survival under different environmental stress conditions suggest that it uses a variety of mechanisms to protect itself against various types of biotic and abiotic stress. R. solanacearum produces a melanin-like brown pigment in the stationary phase when grown in minimal medium containing tyrosine. To gain deeper insight into the genetic determinants involved in melanin production, transposon-inserted mutants of R. solanacearum strain SL341 were screened for strains with defective melanin-producing capability. In addition to one mutant already known to be involved in pyomelanin production (viz., strain SL341D, with disruption of the hydroxphenylpyruvate dioxygenase gene), we identified three other mutants with disruption in the regulatory genes rpoS, hrpG, and oxyR, respectively. Wild-type SL341 produced pyomelanin in minimal medium containing tyrosine whereas the mutant strains did not. Likewise, homogentisate, a major precursor of pyomelanin, was detected in the culture filtrate of the wild-type strain but not in those of the mutant strains. A gene encoding hydroxyphenylpyruvate dioxygenase exhibited a significant high expression in wild type SL341 compared to other mutant strains, suggesting that pyomelanin production is regulated by three different regulatory proteins. However, analysis of the gene encoding homogentisate dioxygenase revealed no significant difference in its relative expression over time in the wild-type SL341 and mutant strains, except for SL341D, at 72 h incubation. The pigmented SL341 strain also exhibited a high tolerance to hydrogen peroxide stress compared with the non-pigmented SL341D strain. Our study suggests that pyomelanin production is controlled by several regulatory factors in R. solanacearum to confer protection under oxidative stress.