The Multifaceted Role of Glutathione S-Transferases in Health and Disease.

In humans, the cytosolic glutathione S-transferase (GST) family of proteins is encoded by 16 genes presented in seven different classes. GSTs exhibit remarkable structural similarity with some overlapping functionalities. As a primary function, GSTs play a putative role in Phase II metabolism by protecting living cells against a wide variety of toxic molecules by conjugating them with the tripeptide glutathione. This conjugation reaction is extended to forming redox sensitive post-translational modifications on proteins: S-glutathionylation. Apart from these catalytic functions, specific GSTs are involved in the regulation of stress-induced signaling pathways that govern cell proliferation and apoptosis. Recently, studies on the effects of GST genetic polymorphisms on COVID-19 disease development revealed that the individuals with higher numbers of risk-associated genotypes showed higher risk of COVID-19 prevalence and severity. Furthermore, overexpression of GSTs in many tumors is frequently associated with drug resistance phenotypes. These functional properties make these proteins promising targets for therapeutics, and a number of GST inhibitors have progressed in clinical trials for the treatment of cancer and other diseases.

Novel Coronavirus (SARS-CoV-2) in Water and Environment-A Scoping Review.

A pneumonia outbreak was primarily reported in the fall of 2019 in Wuhan, Hubei province, China, with the identity SARS-CoV-2, a novel coronavirus. It quickly grew from a local epidemic to a global pandemic and was declared a public health emergency by the WHO. A total of three prominent waves were identified across the globe, with a slight temporal variability as per the geographical locations, and has impacted several sectors which connect the world. By March 2022, the coronavirus had infected 444.12 million people and claimed 6.01 million human lives worldwide, and these numbers have not yet stabilized. Our paper enlightens readers on the seven strains of human coronaviruses, with special emphasis on the three severe deadliest outbreaks (SARS-2002, MERS-2012, and COVID-19). This work attempts a comprehensive understanding of the coronavirus and its impact on the possible sectors that link the world through the economic chain, climate conditions, SDGs, recycling of the event, and mitigations. There are many points that are raised by the authors in the possible sectors, which are emerging or are as yet unnoticed and thus have not been taken into consideration. This comprehension will leave sets of new challenges and opportunities for the researchers in various streams, especially in earth sciences. Science-integrated research may help to prevent upcoming disasters as a by-product of (existing) epidemics in the form of coronavirus.

Risk factors for outbreaks caused by variant strain of Newcastle disease on environmentally controlled broiler chicken farms in Lahore, Pakistan.

Newcastle disease (ND) is a frequently reported disease in poultry among both vaccinated and non-vaccinated flocks in Pakistan. During 2011-2012 poultry industry in Punjab, mainly in Lahore region, faced fatal outbreaks of ND caused by a variant strain. An analytical study was conducted during outbreak period in Lahore region. A total of 114 environmentally controlled farms were selected with the help of convenient sampling method. A questionnaire was designed about the potential risk factors associated with the spread of ND outbreak. The bivariate relationships between ND status and independent variables were investigated by applying the Chi-square and Fisher's exact test. Multivariable logistic model was used to estimate the effect of each studied variable on the outcome by adjusting the other variables in the model. The variables which showed an association with ND outbreaks at commercial poultry farms were improper method for dead birds disposal (OR=4.96; 95% CI 1.63-15.12), use of same feed transporting vehicle at multiple poultry farms (OR=4.92; 95% CI 1.58-15.33), farm to farm distance of less than 1 km (OR=9.32; 95% CI(1.19-73.12), number of sheds at one farm (OR=2.31; 95% CI 0.93-5.69), labor type (OR=2.72; 95% CI 0.83-8.88) and biosecurity (OR= 4.47; 95% CI 0.56-35.66).

Assessing the role of SWIR band in detecting agricultural crop stress: a case study of Raichur district, Karnataka, India.

The potential consequence of global climatic change caused the rise in temperature and precipitation decline, the aftermath of which has led to phenomenon like drought. As the agriculture is mainly dependent on the timely availability of water, delayed arrival of rainfall or decrease in the intensity of precipitation highly affects the growth of crops. If such situation persists for a longer period, the soil moisture content will be exploited completely and maturity of the crop will be stunted, finally adversely affecting the annual crop yield. In the present study, the capability of shortwave-infrared (SWIR) channels in detecting the agricultural crop stress in Raichur district of Karnataka state, India, using spectral vegetation indices namely Global Vegetation Moisture Index (GVMI) and Normalized Multi-band Drought Index (NMDI) has been analyzed using multi-temporal MODIS data. The vegetation health analysis by utilizing both indices were carried out from year 2002 to 2012 for the Kharif season, and it was seen that the year 2002 suffered major agricultural drought where the lower GVMI and NMDI values were covering the majority of the crop areas and in the year 2010 agricultural crop production was observed to be good. The average values of GVMI and NMDI for the years 2002 and 2010 were plotted with the average NDVI values of all months of both years and the results revealed that NDVI values were in concurrence with both NMDI and GVMI.

Potent inhibitors of equine steroid isomerase EcaGST A3-3.

Equine glutathione transferase A3-3 (EcaGST A3-3) belongs to the superfamily of detoxication enzymes found in all higher organisms. However, it is also the most efficient steroid double-bond isomerase known in mammals. Equus ferus caballus shares the steroidogenic pathway with Homo sapiens, which makes the horse a suitable animal model for investigations of human steroidogenesis. Inhibition of the enzyme has potential for treatment of steroid-hormone-dependent disorders. Screening of a library of FDA-approved drugs identified 16 out of 1040 compounds, which at 10 µM concentration afforded at least 50% inhibition of EcaGST A3-3. The most potent inhibitors, anthralin, sennoside A, tannic acid, and ethacrynic acid, were characterized by IC50 values in the submicromolar range when assayed with the natural substrate Δ5-androstene-3,17-dione.

ß-actin regulates a heterochromatin landscape essential for optimal induction of neuronal programs during direct reprograming.

During neuronal development, ß-actin serves an important role in growth cone mediated axon guidance. Consistent with this notion, in vivo ablation of the ß-actin gene leads to abnormalities in the nervous system. However, whether ß-actin is involved in the regulation of neuronal gene programs is not known. In this study, we directly reprogramed ß-actin+/+ WT, ß-actin+/- HET and ß-actin-/- KO mouse embryonic fibroblast (MEFs) into chemically induced neurons (CiNeurons). Using RNA-seq analysis, we profiled the transcriptome changes among the CiNeurons. We discovered that induction of neuronal gene programs was impaired in KO CiNeurons in comparison to WT ones, whereas HET CiNeurons showed an intermediate levels of induction. ChIP-seq analysis of heterochromatin markers demonstrated that the impaired expression of neuronal gene programs correlated with the elevated H3K9 and H3K27 methylation levels at gene loci in ß-actin deficient MEFs, which is linked to the loss of chromatin association of the BAF complex ATPase subunit Brg1. Together, our study shows that heterochromatin alteration in ß-actin null MEFs impedes the induction of neuronal gene programs during direct reprograming. These findings are in line with the notion that H3K9Me3-based heterochromatin forms a major epigenetic barrier during cell fate change.

Expression of a Drosophila glutathione transferase in Arabidopsis confers the ability to detoxify the environmental pollutant, and explosive, 2,4,6-trinitrotoluene.

The explosive 2,4,6-trinitrotoluene (TNT) is a significant, global environmental pollutant that is both toxic and recalcitrant to degradation. Given the sheer scale and inaccessible nature of contaminated areas, phytoremediation may be a viable clean-up approach. Here, we have characterized a Drosophila melanogaster glutathione transferase (DmGSTE6) which has activity towards TNT. Recombinantly expressed, purified DmGSTE6 produces predominantly 2-glutathionyl-4,6-dinitrotoluene, and has a 2.5-fold higher Maximal Velocity (Vmax ), and five-fold lower Michaelis Constant (Km ) than previously characterized TNT-active Arabidopsis thaliana (Arabidopsis) GSTs. Expression of DmGSTE6 in Arabidopsis conferred enhanced resistance to TNT, and increased the ability to remove TNT from contaminated soil relative to wild-type plants. Arabidopsis lines overexpressing TNT-active GSTs AtGST-U24 and AtGST-U25 were compromised in biomass production when grown in the absence of TNT. This yield drag was not observed in the DmGSTE6-expressing Arabidopsis lines. We hypothesize that increased levels of endogenous TNT-active GSTs catalyse excessive glutathionylation of endogenous substrates, depleting glutathione pools, an activity that DmGST may lack. In conclusion, DmGSTE6 has activity towards TNT, producing a compound with potential for further biodegradation. Selecting or manipulating plants to confer DmGSTE6-like activity could contribute towards development of phytoremediation strategies to clean up TNT from polluted military sites.

Food productivity trend analysis of Raichur district for the management of agricultural drought.

Drought is an extreme climatic situation where there is a water shortage arising due to sub-normal rainfall, erratic distribution of precipitation, increased water supply demand, etc. India faced several years of drought in last six decades. As Indian agriculture is largely dependent on the monsoon, a slight change affects production as well as crop yield drastically. Statistical analysis is important for mapping the drought prone areas. Raichur district of the northern interior state of Karnataka is a drought-prone region where the economy is mainly based on agriculture. So, the uneven distribution of rainfall as well as the delay in the arrival of the southwest monsoon adversely affects the growth stage of crops which result in a decline in crop production. The effect of drought on the agriculture for the past decade has been analyzed using crop productivity data. When the production rate of Raichur district was studied for the years 1998 to 2009, it was seen that major crops like rice and jowar faced a decline in its production during the years 2002 and 2003, whereas bajra, maize, etc. mostly decreased in the year 2004.

Drosophila GSTs display outstanding catalytic efficiencies with the environmental pollutants 2,4,6-trinitrotoluene and 2,4-dinitrotoluene.

The nitroaromatic explosive 2,4,6-trinitrotoluene (TNT) and the related 2,4-dinitrotoluene (DNT) are toxic environmental pollutants. The biotransformation and detoxication of these persistent compounds in higher organisms are of great significance from a health perspective as well as for the biotechnological challenge of bioremediation of contaminated soil. We demonstrate that different human glutathione transferases (GSTs) and GSTs from the fruit fly Drosophila melanogaster are catalysts of the biotransformation of TNT and DNT. The human GSTs had significant but modest catalytic activities with both DNT and TNT. However, D. melanogaster GSTE6 and GSTE7 displayed outstanding high activities with both substrates.

Comparison of epsilon- and delta-class glutathione S-transferases: the crystal structures of the glutathione S-transferases DmGSTE6 and DmGSTE7 from Drosophila melanogaster.

Cytosolic glutathione transferases (GSTs) comprise a large family of enzymes with canonical structures that diverge functionally and structurally among mammals, invertebrates and plants. Whereas mammalian GSTs have been characterized extensively with regard to their structure and function, invertebrate GSTs remain relatively unstudied. The invertebrate GSTs do, however, represent potentially important drug targets for infectious diseases and agricultural applications. In addition, it is essential to fully understand the structure and function of invertebrate GSTs, which play important roles in basic biological processes. Invertebrates harbor delta- and epsilon-class GSTs, which are not found in other organisms. Drosophila melanogaster GSTs (DmGSTs) are likely to contribute to detoxication or antioxidative stress during development, but they have not been fully characterized. Here, the structures of two epsilon-class GSTs from Drosophila, DmGSTE6 and DmGSTE7, are reported at 2.1 and 1.5 Šresolution, respectively, and are compared with other GSTs to identify structural features that might correlate with their biological functions. The structures of DmGSTE6 and DmGSTE7 are remarkably similar; the structures do not reveal obvious sources of the minor functional differences that have been observed. The main structural difference between the epsilon- and delta-class GSTs is the longer helix (A8) at the C-termini of the epsilon-class enzymes.

Identification of new inhibitors for human hematopoietic prostaglandin D2 synthase among FDA-approved drugs and other compounds.

OBJECTIVE: Hematopoietic prostaglandin D2 synthase (HPGDS) is a member of the Sigma class glutathione transferases (GSTs) catalyzing the isomerization of prostaglandin H2 to prostaglandin D2, a mediator of allergy and inflammation responses. Selective inhibitors of human HPGDS are expected to be of therapeutic importance in relieving symptoms related to allergy and asthma. Hence, a collection of diverse FDA-approved compounds was screened for potential novel applications as inhibitors of HPGDS. METHODS: The catalytic activity of purified HPGDS was used for inhibition studies in vitro. RESULTS: Our inhibition studies revealed 23 compounds as effective inhibitors of HPGDS with IC50 values in the low micromolar range. Erythrosine sodium, suramin, tannic acid and sanguinarine sulfate were characterized with IC50 values of 0.2, 0.3, 0.4, and 0.6 µM, respectively. Kinetic inhibition analysis showed that erythrosine sodium is a nonlinear competitive inhibitor of HPGDS, while suramin, tannic acid and sanguinarine sulfate are linear competitive inhibitors. CONCLUSION: The results show that certain FDA-approved compounds may have pharmacological effects not previously realized that warrant further consideration in their clinical use.

TLR8 gene polymorphism and association in bacterial load in southern Punjab of Pakistan: an association study with pulmonary tuberculosis.

Only 5-10% of people infected with Mycobacterium tuberculosis develop active tuberculosis which suggests a role of genetic variation in host immunity. Genetic variants in TLRs are potential indicator for host susceptibility and outcome of several diseases. We explored the association of nonsynonymous genetic variants (Met1Val) with Toll-like receptor 8 in Pakistani population. Genotypic and allelic distribution of TLR8 polymorphism (rs3764880) in patients with TB and healthy donors from different areas of southern Punjab, Pakistan, was determined. Results provide that our population is highly influenced by TLR8 Met1Val SNP for TB, and G allele appeared to increase TB susceptibility. Mutant genotype GG or G/- and G allele was significantly higher among all the categories of cases than in controls. Among different levels of bacillary load and genotypes, GG or G/- and G allele significantly supports the incidence of 2 + class for bacterial load.

Overexpression of glutathione transferase E7 in Drosophila differentially impacts toxicity of organic isothiocyanates in males and females.

Organic isothiocyanates (ITCs) are allelochemicals produced by plants in order to combat insects and other herbivores. The compounds are toxic electrophiles that can be inactivated and conjugated with intracellular glutathione in reactions catalyzed by glutathione transferases (GSTs). The Drosophila melanogaster GSTE7 was heterologously expressed in Escherichia coli and purified for functional studies. The enzyme showed high catalytic activity with various isothiocyanates including phenethyl isothiocyanate (PEITC) and allyl isothiocyanate (AITC), which in millimolar dietary concentrations conferred toxicity to adult D. melanogaster leading to death or a shortened life-span of the flies. In situ hybridization revealed a maternal contribution of GSTE7 transcripts to embryos, and strongest zygotic expression in the digestive tract. Transgenesis involving the GSTE7 gene controlled by an actin promoter produced viable flies expressing the GSTE7 transcript ubiquitously. Transgenic females show a significantly increased survival when subjected to the same PEITC treatment as the wild-type flies. By contrast, transgenic male flies show a significantly lower survival rate. Oviposition activity was enhanced in transgenic flies. The effect was significant in transgenic females reared in the absence of ITCs as well as in the presence of 0.15 mM PEITC or 1 mM AITC. Thus the GSTE7 transgene elicits responses to exposure to ITC allelochemicals which differentially affect life-span and fecundity of male and female flies.

Glutathione transferases immobilized on nanoporous alumina: flow system kinetics, screening, and stability.

The previously uncharacterized Drosophila melanogaster Epsilon-class glutathione transferases E6 and E7 were immobilized on nanoporous alumina. The nanoporous anodized alumina membranes were derivatized with 3-aminopropyl-triethoxysilane, and the amino groups were activated with carbonyldiimidazole to allow coupling of the enzymes via ε-amino groups. Kinetic analyses of the immobilized enzymes were carried out in a circulating flow system using CDNB (1-chloro-2,4-dinitrobenzene) as substrate, followed by specificity screening with alternative substrates. A good correlation was observed between the substrate screening data for immobilized enzyme and corresponding data for the enzyme in solution. A limited kinetic study was also carried out on immobilized human GST S1-1 (also known as hematopoietic prostaglandin D synthase). The stability of the immobilized enzymes was virtually identical to that of enzymes in solution, and no leakage of enzyme from the matrix could be observed.

Role of Toll-like receptor 2 (-196 to -174) polymorphism in susceptibility to pulmonary tuberculosis in Pakistani population.

Tuberculosis (TB) is a global public health problem and a source of preventable deaths each year, with 8.8 million new cases of TB and 1.6 million deaths worldwide. Pakistan ranks sixth on the list of 22 high-burden tuberculosis countries in the world. The transitions from infection to clinical disease are very few signifying that host-defence factors could lead to the development of active disease. Toll-like receptor (TLR) 2 polymorphisms have been associated with regulation of TLR expression and development of active TB. In this study, blood samples of 187 subjects including 100 healthy and 87 TB positive were collected from three districts of Pakistan. DNA was extracted from blood and TLR 2 (-196 to -174del) polymorphism was analysed by polymerase chain reaction (PCR). The results suggest that the frequency of -196 to -174del/del polymorphism of TLR2 was significantly higher in TB-positive patients compared with healthy. Results revealed that (-196 to -174del) polymorphism may increase the susceptibility to TB in healthy population of Pakistan. Moreover, males with heterozygous genotype (I/D) are more prone to TB than females with the same genotype. The occurrence of TB infection has been found positively associated with the age, suggesting that the population within the range of 21-45 years is more susceptible to Mycobacterium tuberculosis than other age groups studied. A significant association is also observed between smoking and the chances of developing TB, confirming that smoking strongly promotes its incidence.

A study on the prevalence of a tick-transmitted pathogen, Theileria annulata, and hematological profile of cattle from Southern Punjab (Pakistan).

The present study was carried out to determine the prevalence of Theileria annulata in large ruminants in Southern Punjab (Pakistan). Blood samples were collected from 144 large ruminants, consisting of 105 cattle and 39 buffaloes, from six districts of Southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar, and Vehari. Data on the characteristics of the animals and herds were collected through questionnaires. The age of animals (P = 0.02), presence of ticks on animals (P = 0.02), and presence of ticks on dogs associated with herds (P = 0.05) were among the major risk factors involved in the spread of tropical theileriosis in the study area. Two different parasite detection techniques, PCR amplification and screening of Giemsa-stained slides, were compared, and it was found that PCR amplification is a more sensitive tool (19% parasite detection) as compared to smear scanning (3% parasite detection) for the detection of T. annulata. Twenty eight out of 144 animals produced the 721-bp fragment specific for T. annulata from five out of six sampling districts. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite-positive and parasite-negative samples to assess the effect of T. annulata on the blood and serological profile of infected animals.

Nutlin-3, the small-molecule inhibitor of MDM2, promotes senescence and radiosensitises laryngeal carcinoma cells harbouring wild-type p53.

BACKGROUND: Primary radiotherapy (RT) is a mainstay of treatment for laryngeal squamous cell carcinoma (LSCC). Although the cure rates for early (T1) vocal cord tumours are high, RT proves ineffective in up to a third of T3 carcinomas. Moreover, RT is associated with debilitating early- and late-treatment-related toxicity, thus finding means to de-escalate therapy, while retaining/augmenting therapeutic effectiveness, is highly desirable. p53 is a key mediator of radiation responses; we therefore investigated whether Nutlin-3, a small-molecule inhibitor of MDM2 (mouse double minute 2; an essential negative regulator of p53), might radiosensitise LSCC cells. METHODS: We performed clonogenic assays to measure radiosensitivity in a panel of LSCC cell lines (for which we determined p53 mutational status) in the presence and absence of Nutlin-3. RESULTS: LSCC cells harbouring wild-type p53 were significantly radiosensitised by Nutlin-3 (P<0.0001; log-rank scale), and displayed increased cell cycle arrest and significantly increased senescence (P<0.001) in the absence of increased apoptosis; thus, our data suggest that senescence may mediate this increased radiosensitivity. CONCLUSION: This is the first study showing Nutlin-3 as an effective radiosensitiser in LSCC cells that retain wild-type p53. The clinical application of Nutlin-3 might improve local recurrence rates or allow treatment de-escalation in these patients.

Identification of hepatitis B virus core mutants by PCR-RFLP in chronic hepatitis B patients from Punjab, Pakistan.

Chronic hepatitis B virus (HBV) infection remains a major health issue worldwide. Several factors including core gene variation are responsible for the development of chronicity of HBV infection. The present study was designed to identify the variations in the core region of the HBV genome in a local population of chronic hepatitis B patients (n = 57) using a PCR-based restriction fragment length polymorphism (PCR-RFLP) method. Fifty subjects were found to be positive for the presence of HBV DNA. For the core region genotyping, the Ava II and Msp I restriction enzymes were used. Mutations at nucleotide (nt) 2147 and nt 2362 in the HBV genome in the core region for Ava II (A4 type, 74%) and nt 2331 for Msp I (M1 type, 66%) were observed as the most common pattern. These results are different from those of previously reported studies on other populations and thus appear to be unique to the Pakistani population. This type of characterization of core mutants may be useful for the design of vaccines based on viral epitopes that are effective for the Pakistani population. Moreover, these unique genotypic patterns for the HBV core gene might be some of the main factors responsible for understanding the underlying mechanism by which HBV chronicity is developed in the Pakistani population.