Molecular characterization of enteropathogenic Escherichia coli isolated from patients with gastroenteritis in a tertiary referral hospital of northeast India.

PURPOSE: Diarrhoeal illness accounts for a high morbidity and mortality both in paediatric as well as adult groups and diarrhoeagenic Escherichia coli occupies a top position as a causative agent of infectious diarrhoeal illness worldwide. The aim of the current investigation was to determine the virulence and pattern of antibiotic resistance of enteropathogenic, enterotoxigenic, and shiga toxigenic Escherichia coli that are linked to diarrhoea in patients of both adult and paediatric age groups. METHODS: A total of 50 consecutive, nonduplicate Escherichia coli isolates were collected from patients with gastro-enteritis who were admitted to different clinical wards Silchar Medical College and Hospital, Silchar, India. PCR was used to identify the virulence genes of EPEC (eaeA and bfpA), STEC (stx1, stx2, and eae) and ETEC (eltA, eltB, estA1 and estA2) in the isolates of E. coli. The antibiotic susceptibility pattern of virulent E. coli isolates were checked using disc diffusion method. Molecular typing of the virulent E. coli detected in the study based on enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) was also done. RESULT: Out of 50 E. coli isolates, 13 (26%) were found to carry atleast one virulence gene. 11 isolates harboured eae gene and were characterized as EPEC and two isolates carried stx1 gene of STEC. These virulent isolates showed different antibiotic susceptibility pattern and harboured single or multiple antibiotic resistance genes. ERIC PCR established 12 different clonal patterns of the virulent study isolates of E. coli harbouring. CONCLUSION: EPEC pathotypes were found to be the most detected pathotype in the stool samples. Majority of the virulent isolates were also resistant to multiple antibiotics which is a serious public health concern and therefore requires a proper surveillance and studies to track their reservoirs to contain their spread.

R software for QSAR analysis in phytopharmacological studies.

INTRODUCTION: In recent decades, quantitative structure-activity relationship (QSAR) analysis has become an important method for drug design and natural product research. With the availability of bioinformatic and cheminformatic tools, a vast number of descriptors have been generated, making it challenging to select potential independent variables that are accurately related to the dependent response variable. OBJECTIVE: The objective of this study is to demonstrate various descriptor selection procedures, such as the Boruta approach, all subsets regression, the ANOVA approach, the AIC method, stepwise regression, and genetic algorithm, that can be used in QSAR studies. Additionally, we performed regression diagnostics using R software to test parameters such as normality, linearity, residual histograms, PP plots, multicollinearity, and homoscedasticity. RESULTS: The workflow designed in this study highlights the different descriptor selection procedures and regression diagnostics that can be used in QSAR studies. The results showed that the Boruta approach and genetic algorithm performed better than other methods in selecting potential independent variables. The regression diagnostics parameters tested using R software, such as normality, linearity, residual histograms, PP plots, multicollinearity, and homoscedasticity, helped in identifying and diagnosing model errors, ensuring the reliability of the QSAR model. CONCLUSION: QSAR analysis is vital in drug design and natural product research. To develop a reliable QSAR model, it is essential to choose suitable descriptors and perform regression diagnostics. This study offers an accessible, customizable approach for researchers to select appropriate descriptors and diagnose errors in QSAR studies.

Protective Effects of Pelargonidin against DMBA-Induced Mammary Tumorigenesis in BALB/c Mice through Reduced Oxidative Stress and Lipid Anomalies.

Among luminal types of breast cancers, ER + breast cancer is the most frequently diagnosed cancer globally. ER + breast cancer is commonly treated with SERM drugs that block ER to prevent ER-mediated cancerous growth. Our previous computational screening found pelargonidin (PG) can inhibit ER-signaling with potent bioactivity and satisfactory toxicological features. The present study explored the anti-tumoral prospect of PG against DMBA-induced ER + murine mammary carcinogenesis. The female BALB/c mice were divided into control (A) and DMBA-exposed groups. Following tumor appearance, the DMBA-exposed group was divided into five groups: tumor control, PG-treated (Groups P25, P50, and P100), and tamoxifen-treated (TAM). The results indicated that PG-treatment dose-dependently reduced the mean tumor volume, reinstated body weight loss, and enhanced the percentage survival of tumor-bearing mice. In addition, we recorded a significant reduction in LPO, total cholesterol, and triglycerides and a surge in the activity of antioxidases and phase II detoxifying enzymes in PG-treated animals. PG also dose-dependently increased the serum level of unbound estradiol, an indicator of competitive ER binding by an ER agonist/antagonist. These data suggest that pelargonidin has potent anticancer potential against the animal model of ER + breast cancer that matches the efficiency of tamoxifen with conceivably fewer side effects.

Quorum sensing activities and genomic insights of plant growth-promoting rhizobacteria isolated from Assam tea.

Secretion of quorum sensing (QS) molecules is important for the effective colonization of host plants by plant growth-promoting rhizobacteria. The current study aims at the isolation and characterization of tea rhizo bacteria, which produce the QS molecules, acyl homoserine lactone (AHLs), along with multiple plant growth-promoting (PGP) activities. Thirty-one isolates were isolated from the tea rhizosphere, and screening for PGP activities resulted in the selection of isolates RTE1 and RTE4 with multiple PGP traits, inhibiting the growth of tea fungal pathogens. Both isolates also showed production of AHL molecules when screened using two biosensor strains, Chromobacterium violaceum CV026 and Escherichia coli MT 102(jb132). The isolates identified as Burkholderia cepacia RTE1 and Pseudomonas aeruginosa RTE4 based on genome-based analysis like phylogeny, dDDH, and fastANI calculation. Detailed characterization of AHLs produced by the isolates using reverse-phase TLC, fluorometry, and LC-MS indicated that the isolate RTE1 produced a short chain, C8, and a long chain C12 AHL, while RTE4 produced short-chain AHLs C4 and C6. Confocal microscopy revealed the formation of thick biofilm by RTE1 and RTE4 (18 and 23 µm, respectively). Additionally, we found several genes involved in QS, and PGP, inducing systemic resistance (ISR) activities such as lasI/R, qscR, pqq, pvd, aldH, acdS, phz, Sod, rml, and Pch, and biosynthetic gene clusters like N-acyl homoserine lactone synthase, terpenes, pyochelin, and pyocyanin. Based on the functional traits like PGP, biofilm formation and production of AHL molecules, and genetic potential of the isolates B. cepacia RTE1 and P. aeruginosa RTE4 appear promising candidates to improve the health and growth of tea plantations.

Hibiscus sabdariffa anthocyanins are potential modulators of estrogen receptor alpha activity with favourable toxicology: a computational analysis using molecular docking, ADME/Tox prediction, 2D/3D QSAR and molecular dynamics simulation.

The estrogen hormone receptor (ER) mediated gene expression mainly regulate the development and physiology of the primary and secondary reproductive system alongside bone-forming, metabolism and behaviour. Over-expressed ER is associated with several pathological conditions and play a crucial role in breast cancer occurrence, progression and metastasis. Hibiscus sabdariffa L. or roselle is a rich source of naturally occurring polyphenolic compounds that reportedly have robust estrogenic activity. However, the estrogen-like ingredient of the plant remains ambiguous. This study has screened a library of already recorded and less-explored compounds of Hibiscus sabdariffa for their estrogen receptor binding affinity and safety using a suite of computational methods that include protein-ligand docking, ADME and Toxicity prediction, and 2D/3D QSAR. The study revealed that the estrogen-receptor binding potential of Pelargonidin, Delphinidin, Cyanidin, and Hibiscetin are more efficient than popular breast cancer drugs, Tamoxifen and Raloxifene. Besides, the compounds exhibited favourable toxicological parameters with potent bioactivity towards binding ER-α subunit. Thus, these compounds can serve as prototypes for designing novel Selective Estrogen Receptor Modulator molecules with a few structural modifications. This is the first report exploring the phytochemical basis of estrogenic activity of Hibiscus sabdariffa L.Communicated by Ramaswamy H. Sarma.

Helping plants to deal with heavy metal stress: the role of nanotechnology and plant growth promoting rhizobacteria in the process of phytoremediation.

Heavy metals (HMs) are not destroyable or degradable and persist in the environment for a long duration. Thus, eliminating and counteracting the HMs pollution of the soil environment is an urgent task to develop a safe and sustainable environment. Plants are in close contact with the soil and can play an important role in soil clean-up, and the process is known as phytoremediation. However, under HM contaminated conditions, plants suffer from several complications, like nutrient and mineral deficiencies, alteration of various physiological and biological processes, which reduces the plant's growth rate. On the other hand, the bioavailability of HMs is another factor for reduced phytoremediation, as most of the HMs are not bioavailable to plants for efficient phytoremediation. The altered plant growth and reduced bioavailability of HMs could be overcome and enhance the phytoremediation efficiency by incorporating either nanotechnology, i.e., nanoparticles (NPs) or plant growth promoting rhizobacteria (PGPR) along with phytoremediation. Single incorporation of NPs and PGPR might improve the growth rate in plants by enhancing nutrient availability and uptake and also by regulating plant growth regulators under HM contaminated conditions. However, there are certain limitations, like a high dose of NPs that might have toxic effects on plants. Thus, the combination of two techniques such as PGPR and NPs-based remediation can conquer the limitations of individual techniques and consequently enhance phytoremediation efficiency. Considering the negative impacts of HMs on the environment and living organisms, this review is aimed at highlighting the concept of phytoremediation, the single or combined integration of NPs and PGPR to help plants deal with HMs and their basic mechanisms involved in the process of phytoremediation. Additionally, the complications of using NPs and PGPR in the phytoremediation process are discussed to determine future research questions and this will assist to stimulate further research in this field and increase its effectiveness in practical application.

Antioxidant enzymatic activities and profiling of gene expression associated with organophosphate stress tolerance in Solanum melongena L.cv. Longai.

The tolerance mechanism of chemical pesticide is necessary to combat the pest infestation challenges. This study intended to analyze the responses of enzymatic activity and expression level of an antioxidant gene to organophosphate pesticide stress. The alteration of anti-oxidative correlated with pesticide treatment in eggplant (S. melongena L.cv. Longai) using varying concentrations (0, 50, 100, 150 and 200 ppm) of malathion (PM) and tatafen (PTF) each. The enzyme activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were observed to be elevated with pesticide treatment in eggplant seedling. FeSOD (iron SOD), CAT and APX genes associated in defense mechanisms were significantly expressed under PM and PTF stress which contributed to stress tolerance to the plant. The different concentration of both pesticide stresses altered the expression level of mRNA, FeSOD, CAT and APX genes in comparison to the non-treated plant. While mRNA level of three antioxidant genes were evaluated and found to be APX gene expression was more potent than the CAT and FeSOD gene subjected to different concentrations of PM and PTF in eggplant. The current experiment highlights the presence of minimum level of pesticide concentration impacted positively towards the plant growth and metabolism, while high level of pesticide concentration impacted negatively. In summary, antioxidant enzymes activity responded to both pesticide stresses at an early stage of exposure and their gene expression profiles provided more details about their complex interaction and effectively scavenge reactive oxygen species. This allows the plant to maintain growth under pesticide stress.

The Endophytic Microbiome as a Hotspot of Synergistic Interactions, with Prospects of Plant Growth Promotion.

The plant root is the primary site of interaction between plants and associated microorganisms and constitutes the main components of plant microbiomes that impact crop production. The endophytic bacteria in the root zone have an important role in plant growth promotion. Diverse microbial communities inhabit plant root tissues, and they directly or indirectly promote plant growth by inhibiting the growth of plant pathogens, producing various secondary metabolites. Mechanisms of plant growth promotion and response of root endophytic microorganisms for their survival and colonization in the host plants are the result of complex plant-microbe interactions. Endophytic microorganisms also assist the host to sustain different biotic and abiotic stresses. Better insights are emerging for the endophyte, such as host plant interactions due to advancements in 'omic' technologies, which facilitate the exploration of genes that are responsible for plant tissue colonization. Consequently, this is informative to envisage putative functions and metabolic processes crucial for endophytic adaptations. Detection of cell signaling molecules between host plants and identification of compounds synthesized by root endophytes are effective means for their utilization in the agriculture sector as biofertilizers. In addition, it is interesting that the endophytic microorganism colonization impacts the relative abundance of indigenous microbial communities and suppresses the deleterious microorganisms in plant tissues. Natural products released by endophytes act as biocontrol agents and inhibit pathogen growth. The symbiosis of endophytic bacteria and arbuscular mycorrhizal fungi (AMF) affects plant symbiotic signaling pathways and root colonization patterns and phytohormone synthesis. In this review, the potential of the root endophytic community, colonization, and role in the improvement of plant growth has been explained in the light of intricate plant-microbe interactions.

Pseudomonas aeruginosa RTE4: A Tea Rhizobacterium With Potential for Plant Growth Promotion and Biosurfactant Production.

Tea is an ancient non-alcoholic beverage plantation crop cultivated in the most part of Assam, India. Being a long-term monoculture, tea plants are prone to both biotic and abiotic stresses, and requires massive amounts of chemicals as fertilizers and pesticides to achieve worthy crop productivity. The rhizosphere bacteria with the abilities to produce phytohormone, secreting hydrolytic enzyme, biofilm formation, bio-control activity provides induced systemic resistance to plants against pathogens. Thus, plant growth promoting (PGP) rhizobacteria represents as an alternative candidate to chemical inputs for agriculture sector. Further, deciphering the secondary metabolites, including biosurfactant (BS) allow developing a better understanding of rhizobacterial strains. The acidic nature of tea rhizosphere is predominated by Bacillus followed by Pseudomonas that enhances crop biomass and yield through accelerating uptake of nutrients. In the present study, a strain Pseudomonas aeruginosa RTE4 isolated from tea rhizosphere soil collected from Rosekandy Tea Garden, Cachar, Assam was evaluated for various plant-growth promoting attributes. The strain RTE4 produces indole acetic acid (74.54 µg/ml), hydrolytic enzymes, and solubilize tri-calcium phosphate (46 µg/ml). Bio-control activity of RTE4 was recorded against two foliar fungal pathogens of tea (Corticium invisium and Fusarium solani) and a bacterial plant pathogen (Xanthomonas campestris). The strain RTE4 was positive for BS production in the preliminary screening. Detailed analytical characterization through TLC, FTIR, NMR, and LCMS techniques revealed that the strain RTE4 grown in M9 medium with glucose (2% w/v) produce di-rhamnolipid BS. This BS reduced surface tension of phosphate buffer saline from 71 to 31 mN/m with a critical micelle concentration of 80 mg/L. Purified BS of RTE4 showed minimum inhibitory concentration of 5, 10, and 20 mg/ml against X. campestris, F. solani and C. invisium, respectively. Capability of RTE4 BS to be employed as a biofungicide as compared to Carbendazim - commercially available fungicide is also tested. The strain RTE4 exhibits multiple PGP attributes along with production of di-rhamnolipid BS. This gives a possibility to produce di-rhamnolipid BS from RTE4 in large scale and explore its applications in fields as a biological alternative to chemical fertilizer.

Linking gut microbiota with the human diseases.

The human gut is rich in microbes. Therefore, it is of interest to document data to link known human diseases with the gut microbiota. Various factors like hormones, metabolites and dietary habitats are responsible for shaping the microbiota of the gut. Imbalance in the gut microbiota is responsible for the pathogenesis of various disease types including rheumatoid arthritis, different types of cancer, diabetes mellitus, obesity, and cardiovascular disease. We report a review of known data for the correction of dysbiosis (imbalance in microbe population) towards improved human health.

Insight into the molecular evidence supporting the remarkable chemotherapeutic potential of Hibiscus sabdariffa L.

Hibiscus sabdariffa or roselle tea is popular around the globe for its antioxidant capability along with various other health benefits. Besides, it has uses in Ayurvedic and Chinese herbal medicines for the treatment of several diseases. However, the investigation for the anticancer potential of the plant began roughly in the last decade that emerged with encouraging results. Both crude extracts and pure compounds of the plant were reported to induce chemoprevention, selective cytotoxicity, cell cycle arrest, apoptosis, autophagy and anti-metastasis effects in varied types of human cancer cells. The plant contains a high quantity of polyphenolic compounds and at least two of them were proven to induce potent anticancer effects. Although, the molecular mechanism underlying the anticancer activity was roughly estimated in several studies. The present review aimed to assemble all ambiguous information to report the molecular evidence establishing the potent anticancer activity of Hibiscus sabdariffa and its implication for cancer therapy. This study suggests that Hibiscus sabdariffa is an ideal candidate to investigate its role as a herbal supplement for cancer prevention and treatment. With excellent safety and tolerability record, polyphenolic compounds from the plant need better designed clinical trials.

Bioethanol production from waste lignocelluloses: A review on microbial degradation potential.

Tremendous explosion of population has led to about 200% increment of total energy consumptions in last twenty-five years. Apart from conventional fossil fuel as limited energy source, alternative non-conventional sources are being explored worldwide to cater the energy requirement. Lignocellulosic biomass conversion for biofuel production is an important alternative energy source due to its abundance in nature and creating less harmful impacts on the environment in comparison to the coal or petroleum-based sources. However, lignocellulose biopolymer, the building block of plants, is a recalcitrant substance and difficult to break into desirable products. Commonly used chemical and physical methods for pretreating the substrate are having several limitations. Whereas, utilizing microbial potential to hydrolyse the biomass is an interesting area of research. Because of the complexity of substrate, several enzymes are required that can act synergistically to hydrolyse the biopolymer producing components like bioethanol or other energy substances. Exploring a range of microorganisms, like bacteria, fungi, yeast etc. that utilizes lignocelluloses for their energy through enzymatic breaking down the biomass, is one of the options. Scientists are working upon designing organisms through genetic engineering tools to integrate desired enzymes into a single organism (like bacterial cell). Studies on designer cellulosomes and bacteria consortia development relating consolidated bioprocessing are exciting to overcome the issue of appropriate lignocellulose digestions. This review encompasses up to date information on recent developments for effective microbial degradation processes of lignocelluloses for improved utilization to produce biofuel (bioethanol in particular) from the most plentiful substances of our planet.

Microbial Beta Glucosidase Enzymes: Recent Advances in Biomass Conversation for Biofuels Application.

The biomass to biofuels production process is green, sustainable, and an advanced technique to resolve the current environmental issues generated from fossil fuels. The production of biofuels from biomass is an enzyme mediated process, wherein ß-glucosidase (BGL) enzymes play a key role in biomass hydrolysis by producing monomeric sugars from cellulose-based oligosaccharides. However, the production and availability of these enzymes realize their major role to increase the overall production cost of biomass to biofuels production technology. Therefore, the present review is focused on evaluating the production and efficiency of ß-glucosidase enzymes in the bioconversion of cellulosic biomass for biofuel production at an industrial scale, providing its mechanism and classification. The application of BGL enzymes in the biomass conversion process has been discussed along with the recent developments and existing issues. Moreover, the production and development of microbial BGL enzymes have been explained in detail, along with the recent advancements made in the field. Finally, current hurdles and future suggestions have been provided for the future developments. This review is likely to set a benchmark in the area of cost effective BGL enzyme production, specifically in the biorefinery area.

Methanolic Extract of Curcuma caesia Roxb. Prevents the Toxicity Caused by Cyclophosphamide to Bone Marrow Cells, Liver and Kidney of Mice.

INTRODUCTION: With an ever increasing cause of cancer, it has been recommended to treat with conventional drugs, however because of the side effects caused by the conventional drugs, the research on medicinal plants has been intensified due to their less adverse and toxic effects. OBJECTIVES: The primary objective of the present study was to evaluate the protective effect of the medicinal plant Curcuma caesia Roxb. against free radicals ABTS(+) and O2 (-). Also it was aimed to evaluate the protective effect of C.caesia Roxb. against the chemotherapeutic drug Cyclophosphamide and its side effects in liver and kidney. METHODS: The rhizomes of the plant was extracted with methanol through soxhlet and its antioxidant activity was tested against ABTS(+) and O2 (-). For antigenotoxic studies, animals were divided into eight groups and micronucleus assay was employed and for biochemical analysis serum sample was collected from the blood and SGOT, SGPT analysis was performed. Also the biochemical analysis was performed from both the liver and kidney. RESULTS: The methanolic extract of Curcuma caesia Roxb. was found to scavenge the free radicals ABTS(+) and O2 (-). the micronuclei formation was found to be increased in the positive control group as compared to the negative control group significantly (P<0.002) however increase in the number of micronuclei was found to be decrease with the pretreatment of the extract at different concentrations significantly as compared to the negative control groups (P<0.01, P<0.005, P<0.001). The increased level of serum SGPT and SGOT as well as peroxidation level in both liver and kidney due to treatment of cyclophosphamide was also found to be decreased with the pretreatment of the extract significantly as compared to the positive control groups. There was decreased in the level of endogenous antioxidant such as GSH and GR in the positive control group however decreased level of GSH and GR was found to be increased with the pretreatment of the methanolic extract of C. caesia Roxb. CONCLUSION: The present study suggested that the methanolic extract of C. caesia Roxb has not shown any genotoxicity and reduces the genotoxicity caused by cyclophosphamide. It was also to have the protective effects against the liver and kidney. So it could be provided as one of the herbal supplementation in chemoprevention of CP to ameliorate the side effects of it. SUMMARY: Cancer is characterized by uncontrolled growth of cells and much research has been done for the past several years from various disciplines for the treatment of cancer but till now no therapy has been discovered. Treatment of cancer with chemotherapeutic drugs has been suggested to prevent cancer cells however they are often limited with their toxicity to normal cells. Therefore it has been suggested that the supplementation of medicinal plants which are rich source of antioxidants can decrease the toxic effect caused by chemotherapeutic drugs. Curcuma caesia Roxb is a medicinal plant which has high antioxidant activity, as per present study, methanolic extract of Curcuma caesia Roxb prevents the toxicity caused by cyclophosphosphamide (chemotherapeutic drug) in bone marrow cells by reducing the micronuclei formation; it also prevents the hepatotoxicity and nephrotoxicity caused by cyclophosphamide, so it can be used as a supplement in cancer treatment with cyclophosphamide.

Regulation of p53-targeting microRNAs by polycyclic aromatic hydrocarbons: Implications in the etiology of multiple myeloma.

Multiple myeloma (MM) is a common and deadly cancer of blood plasma cells. A unique feature of MM is the extremely low somatic mutation rate of the p53 tumor suppressor gene, in sharp contrast with about half of all human cancers where this gene is frequently mutated. Eleven miRNAs have been reported to repress p53 through direct interaction with the 3' untranslated region. The expression of nine of them is higher in MM plasma cells than in healthy donor counterparts, suggesting that miRNA overexpression is responsible for p53 inactivation in MM. Here, we report that the environmental carcinogen benzo[a]pyrene (BaP) upregulated the expression of seven p53-targeting miRNAs (miR-25, miR-15a, miR-16, miR-92, miR-125b, miR-141, and miR-200a), while 2,3,7,8-tetrachlorodibenzo-ρ-dioxin (TCDD) upregulated two of them (miR-25 and miR-92) in MM cells. The miR-25 promoter was activated by both BaP and TCDD, and this response was mediated by the aryl hydrocarbon receptor (AhR). We screened 727 compounds that inhibit MM cell survival and down-regulate the expression of p53-targeting miRNAs. We found that (-)-epigallocatechin-3-gallate (EGCG), a constituent of green tea and a major component of the botanical drug Polyphenon® E, reduced the expression of four p53-targeting miRNAs, including miR-25, miR-92, miR-141, and miR-200a. Collectively, these data implicate polycyclic aromatic hydrocarbons and AhR in the regulation of p53-targeting miRNAs in MM and identify a potential therapeutic and preventive agent to combat this deadly disease.