Dramatic increase in antimicrobial resistance in ESKAPE clinical isolates over the 2010-2020 decade in India.

RATIONALE AND OBJECTIVES: ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) constitute a threat to humans worldwide. India is now the most populous country. The goal was to investigate the evolution of the rates of antimicrobial resistance in ESKAPE pathogens across India over the 2010-20 decade. METHODS: The data (89 studies) were retrieved from the Medline PubMed repository using specific keywords. RESULTS: The study of 20 177 ESKAPE isolates showed that A. baumannii isolates were the most represented (35.9%, n = 7238), followed by P. aeruginosa (25.3%, n = 5113), K. pneumoniae (19.5%, n = 3934), S. aureus (16.3%, n = 3286), E. faecium (2.6%, n = 517) and Enterobacter spp. (0.4%, n = 89). A notable increase in the resistance rates to antimicrobial agents occurred over the 2010-20 decade. The most important levels of resistance were observed in 2016-20 for A. baumannii (90% of resistance to the amoxicillin-clavulanate combination) and K. pneumoniae (81.6% of resistance to gentamycin). The rise in ß-lactamase activities was correlated with an increase in the positivity of Gram-negative isolates for ß-lactamase genes. CONCLUSIONS: This review highlighted that, in contrast to developed countries that kept resistance levels under control, a considerable increase in resistance to various classes of antibiotics occurred in ESKAPE pathogens in India over the 2010-2020 decade.

Antibiotic resistance: a global crisis, problems and solutions.

Healthy state is priority in today's world which can be achieved using effective medicines. But due to overuse and misuse of antibiotics, a menace of resistance has increased in pathogenic microbes. World Health Organization (WHO) has announced ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) as the top priority pathogens as these have developed resistance against certain antibiotics. To combat such a global issue, it is utmost important to identify novel therapeutic strategies/agents as an alternate to such antibiotics. To name certain antibiotic adjuvants including: inhibitors of beta-lactamase, efflux pumps and permeabilizers for outer membrane can potentially solve the antibiotic resistance problems. In this regard, inhibitors of lytic domain of lytic transglycosylases provide a novel way to not only act as an alternate to antibiotics but also capable of restoring the efficiency of previously resistant antibiotics. Further, use of bacteriophages is another promising strategy to deal with antibiotic resistant pathogens. Taking in consideration the alternatives of antibiotics, a green synthesis nanoparticle-based therapy exemplifies a good option to combat microbial resistance. As horizontal gene transfer (HGT) in bacteria facilitates the evolution of new resistance strains, therefore identifying the mechanism of resistance and development of inhibitors against it can be a novel approach to combat such problems. In our perspective, host-directed therapy (HDT) represents another promising strategy in combating antimicrobial resistance (AMR). This approach involves targeting specific factors within host cells that pathogens rely on for their survival, either through replication or persistence. As many new drugs are under clinical trials it is advisable that more clinical data and antimicrobial stewardship programs should be conducted to fully assess the clinical efficacy and safety of new therapeutic agents.

Treatment of industrial wastewaters by algae-bacterial consortium with Bio-H2 production: Recent updates, challenges and future prospects.

Currently, scarcity/security of clean water and energy resources are the most serious problems worldwide. Industries use large volume of ground water and a variety of chemicals to manufacture the products and discharge large volume of wastewater into environment, which causes severe impacts on environment and public health. Fossil fuels are considered as major energy resources for electricity and transportation sectors, which release large amount of CO2 and micro/macro pollutants, leading to cause the global warming and public health hazards. Therefore, algae-bacterial consortium (A-BC) may be eco-friendly, cost-effective and sustainable alternative way to treat the industrial wastewaters (IWWs) with Bio-H2 production. A-BC has potential to reduce the global warming and eutrophication. It also protects environment and public health as it converts toxic IWWs into non or less toxic (biomass). It also reduces 94%, 90% and 50% input costs of nutrients, freshwater and energy, respectively during IWWs treatment and Bio-H2 production. Most importantly, it produce sustainable alternative (Bio-H2) to replace use of fossil fuels and fill the world's energy demand in eco-friendly manner. Thus, this review paper provides a detailed knowledge on industrial wastewaters, their pollutants and toxic effects on water/soil/plant/humans and animals. It also provides an overview on A-BC, IWWs treatment, Bio-H2 production, fermentation process and its enhancement methods. Further, various molecular and analytical techniques are also discussed to characterize the A-BC structure, interactions, metabolites and Bio-H2 yield. The significance of A-BC, recent update, challenges and future prospects are also discussed.

Knowledge and Attitude toward Antibiotic Use and Identification of Financially Feasible Options to Curb the Spread of Antibiotics in Environment.

A survey on antibiotic literacy in terms of the use and abuse of antibiotics to track and understand antibiotic consumption is crucial to optimize the use of antibiotics and minimizing antimicrobial resistance (AMR). Purposive random sampling, using the snow-ball questionnaire technique, was adopted to ensure that the respondents distributed across India, coming from rural and urban settings, were adolescents as well as adults and had completed at least the higher secondary school level of education. Respondents were divided into five subcategories. The questionnaire was distributed between April 2021 and July 2021, during the second COVID-19 wave in India. The survey questionnaire included 34 questions, comprising multiple-choice and 5-point Likert scale-type questions. This study composed of 972 respondents. Most respondents considered antibiotics safe and frequently failed to discriminate between the symptoms of bacterial and viral infections, most often leading to self-prescription. About 34% of the rural participants and 50% of the urban participants considered antibiotic resistance a serious health concern. Antibiotic prescriptions by the medical or paramedical practitioner were largely empirical. At least 95% of participants acknowledged having heard about antibiotics; nearly 20% of antibiotic consumption came from nonprescription users, while 30% had not completed their antibiotic therapy for a variety of reasons. Sixty-two percent consumed antibiotics to treat cold and flu symptoms. Results from the survey suggest the presence of a crucial gap between the respondents' perception of antibiotics and levels of information regarding antibiotic use and misuse. The present study may serve as a benchmark that strongly recommends a financially feasible policy, which includes educating society regarding the spread of AMR and its severe consequences by incorporating AMR into the curriculum at the levels of senior secondary school and higher education.

Could Vitamins Have a Positive Impact on the Treatment of Parkinson's Disease?

Parkinson's disease (PD) is the second most common progressive neurodegenerative disorder after Alzheimer's disease. Pathophysiologically, it is characterized by intracytoplasmic aggregates of α-synuclein protein in the Lewy body and loss of dopaminergic neurons from substantia nigra pars compacta and striatum regions of the brain. Although the exact mechanism of neurodegeneration is not fully elucidated, it has been reported that environmental toxins such as MPTP, rotenone, paraquat, and MPP+ induce oxidative stress, which is one of the causative factors for it. To date, there is no complete cure. However, the indispensable role of oxidative stress in mediating PD indicates that antioxidant therapy could be a possible therapeutic strategy against the disease. The deficiency of vitamins has been extensively co-related to PD. Dietary supplementation of vitamins with antioxidant, anti-inflammatory, anti-apoptotic, and free radical scavenging properties could be the potential neuroprotective therapeutic strategy. This review summarizes the studies that evaluated the role of vitamins (A, B, C, D, E, and K) in PD. It will guide future studies in understanding the potential therapeutic role of vitamins in disease pathophysiology and may provide a framework for designing treatment strategies against the disease.

Co-existence of Extended Spectrum ß-Lactamase and carbapenemase-producing genes from Diarrheagenic Enteric pathogens isolated in a tertiary care hospital.

Multiple drug resistance (MDR) among bacterial pathogens is a growing concern that clinicians are facing worldwide. Diarrhea among infants is frequent and is caused by various bacterial and viral infectious agents. Two hundred and twelve stool specimens were collected from pediatric patients from a rural quaternary hospital in Barshi, Sholapur, India, between March and December 2017. Total 180 specimens were positive for various bacterial pathogens, while the remaining 32 diarrhea cases may have been caused by a viral or uncultured bacterial pathogen. Identification of the bacterium and its antibiotic susceptibility were primarily carried out with VITEK-2. Distribution of diarrhea-causing bacteria among the 180 samples was as follows: 61.11% (110) Escherichia coli, 30.55% (55) Klebsiella pneumoniae, 4.44% (8) Proteus mirabilis, 2.22% (4) Shigella spp. 1.11% (2) Morganella morganii and 0.55% (1) each for Enterobacter cloacae and Citrobacter koseri. There was a co-existence of multiple genetic traits conferring extreme drug resistance (XDR) status to 19 isolates, 17 of which were determined to be E. coli and one each of E. cloacae and C. koseri. Antibiotype determination using VITEK-2 and polymerase chain reaction (PCR) amplification of the genetic traits indicated the co-existence of blaTEM and blaCTX-M15 isolates in all 19 isolates, with the exception of E. cloacae. Results showed that 10 out of 19 strains expressed the AmpC cephalosporinase blaCMY-2 gene, whereas metallo-carbapenemase was expressed in four isolates. Distribution of blaNDM-11 and acquired penicillinase blaSHV-1 resistance among 180 clinical isolates is discussed in the light of ESBL traits. This is the first report from the rural part of Maharashtra India showing that as many as 10.55% of the pathogenic strains were XDR, a step ahead of MDR.

Green synthesis and characterization of Solanum xanthocarpum capped silver nanoparticles and its antimicrobial effect on multidrug-resistant bacterial (MDR) isolates.

Plant extracts and their bioactive compounds are considered as the promising options for green synthesis of nanoparticles instead expensive and hazardous materials. Here, Solanum xanthocarpum fruit was used for synthesis of silver nanoparticles (AgNP). The synthesized AgNPs were characterized by using chromatographic and spectroscopic analytical methods. AgNPs were confirmed by UV-visible absorbance at 420-470 nm. TEM analysis showed AgNP with 22.45 nm average size. X-ray diffraction studies revealed the crystalline and face central cubic nature of AgNPs. FTIR analysis revealed functional group present over AgNPs. The aminodiphenyl acetic acid, clomipramine, and fonisopril from fruit extracts were found to be major capping agents on AgNPs as a result of analysis by HRLC-MS. All clinical isolates showed resistance for ampicilline, amoxyclav, niladixic acid, and sulphafurazole, suggesting multidrug resistance. The results showed that all isolates were sensitive to AgNPs synthesized fruit extracts. On the contrary, all isolates were resistant to whole S. xanthocarpum fruit extracts alone. The antimicrobial activity of AgNP was explored against multidrug-resistant (MDR) Gram-negative clinical isolates including Escherichia coli, Shigella spp., Aeronomonas spp. and Pseudomonas spp. MIC values ranged between 1.25 mg/ml and 2.5 mg/ml at 8 McFarland's standards. Minimum bactericidal concentration was found to be in between 2.5 mg/ml to 5 mg/ml. Nanoparticles synthesized from fruit extract of S. xanthocarpum containing aminodiphenyl acetic acid, clomipramine, and fonisopril metabolites exhibit promising antimicrobial activity against MDR Gram-negative clinical isolates.

Subtype diversity and emergence of drug resistance in HIV-1 in solapur district of Maharashtra, India.

Background and Objectives: Even after four decades, HIV infection remains a global challenge and a leading cause of mortality in adults across the world. Anti-retroviral therapy (ART) that controls HIV viremia, is now available through public health facilities in India but drug resistance, which is likely to develop among these individuals remains poorly studied in India. The objectives of present study are to find out the HIV-1 virus subtypes, drug resistance mutations and HIV-1 drug resistance to NRTI, NNRTI and protease inhibitors in the Solapur district, India. Materials and Methods: In a cross sectional study, forty two ART-experienced HIV-1-infected patients with CD4+ count < 200 cells ml-1 and viral load (VL) > 3, 000 copies ml-1 were recruited. All patients belonged to Maharashtra State of India near Barshi Solapur and had been on ART treatment for over 5 years. EDTA whole blood from HIV-1-infected patients was centrifuged and the viral nucleic acid was purified from the plasma. Viral nucleic acid was amplified by PCR using protease and reverse transcriptase specific primers. The resulting amplicons were sequenced and studied for mutations. The tools from Stanford University website were used for subtyping of HIV-1 and identification of mutations conferring drug resistance. Results: In present investigation, HIV-1 subtypes were subtype C in 37 (88.09%), subtype CRF01_AE in 2 (4.76%), and subtype A in 3 patients (7.14%). Drug resistance mutations of NRTI, NNRTI and protease were observed in 15 (37.71%) of 42 patients tested. Drug resistance for NRTI was observed in 12 (28.57%) and for NNRTI in 13 (30.95%) patients. No drug resistance was observed for protease inhibitors. Conclusion: Considerable HIV-1 drug resistance exists among patients receiving ART from a rural areas of India, suggesting more studies from rural region are required to prevent development of resistance to ART.

Green Synthesis of Silver Nanoparticles Using the Tridax procumbens Plant Extract and Screening of Its Antimicrobial and Anticancer Activities.

In this study, we report the green synthesis of silver nanoparticles (AgNPs) using the aqueous leaf extract of Tridax procumbens (TNP), which acts as the source of the reducing and capping agent. The distinctive absorption at 370 nm suggested synthesis of TNPs, which was confirmed by TEM, with a size in the range of 11.1 nm to 45.4 nm and a spherical shape, having a face-centered cubic structure, analyzed by XRD, and a Zeta potential of -20.7 mV, which indicated a moderate stability of TNP. The FTIR analysis revealed the presence of amines and hydroxyl groups with fluoro compounds over the TNPs. The HRLC-MS analysis of TNPs suggested the presence of a major capping agent such as fosinopril and reducing agents such as peptides (Gln Gly Ala, Ser Pro Asn, and Leu Met), terpenoids (lupanyl acid, tiamulin), polyphenol (peucenin), and alkaloids (8',10'-dihydroxydihydroergotamine, carteolol). The synthesized silver nanoparticles exhibited antimicrobial activity against multidrug-resistant (MDR) clinical isolates (Escherichia coli, Shigella spp., Aeromonas spp., Pseudomonas aeruginosa, and Candida tropicalis) and had anticancer activity against A459 (IC50 42.70 µg/ml). The extraction of partially purified aqueous leaf extracts by silica gel column chromatography followed by HPLC to synthesize silver nanoparticles (TNP11) and analyzed by HRLC-MS suggested that dipeptides were involved in the reduction of Ag+ to Ag0. Overall, the results showed that the green silver nanoparticles of T. procumbens could be safe, as they are endowed with potential antimicrobial activity against MDR clinical isolates and human lung carcinoma cells.

In-Vitro Selection of Ceftazidime/Avibactam Resistance in OXA-48-Like-Expressing Klebsiella pneumoniae: In-Vitro and In-Vivo Fitness, Genetic Basis and Activities of ß-Lactam Plus Novel ß-Lactamase Inhibitor or ß-Lactam Enhancer Combinations.

Ceftazidime/avibactam uniquely demonstrates activity against both KPC and OXA-48-like carbapenemase-expressing Enterobacterales. Clinical resistance to ceftazidime/avibactam in KPC-producers was foreseen in in-vitro resistance studies. Herein, we assessed the resistance selection propensity of ceftazidime/avibactam in K. pneumoniae expressing OXA-48-like ß-lactamases (n = 10), employing serial transfer approach. Ceftazidime/avibactam MICs (0.25-4 mg/L) increased to 16-256 mg/L after 15 daily-sequential transfers. The whole genome sequence analysis of terminal mutants showed modifications in proteins linked to efflux (AcrB/AcrD/EmrA/Mdt), outer membrane permeability (OmpK36) and/or stress response pathways (CpxA/EnvZ/RpoE). In-vitro growth properties of all the ceftazidime/avibactam-selected mutants were comparable to their respective parents and they retained the ability to cause pulmonary infection in neutropenic mice. Against these mutants, we explored the activities of various combinations of ß-lactams (ceftazidime or cefepime) with structurally diverse ß-lactamase inhibitors or a ß-lactam enhancer, zidebactam. Zidebactam, in combination with either cefepime or ceftazidime, overcame ceftazidime/avibactam resistance (MIC range 0.5-8 mg/L), while cefepime/avibactam was the second best (MIC: 0.5-16 mg/L) in yielding lower MICs. The present work revealed the possibility of ceftazidime/avibactam resistance in OXA-48-like K. pneumoniae through mutations in proteins involved in efflux and/or porins without concomitant fitness cost mandating astute monitoring of ceftazidime/avibactam resistance among OXA-48 genotypes.

Retrospective Analysis on Antimicrobial Resistance Trends and Prevalence of ß-lactamases in Escherichia coli and ESKAPE Pathogens Isolated from Arabian Patients during 2000-2020.

The production of diverse and extended spectrum ß-lactamases among Escherichia coli and ESKAPE pathogens is a growing threat to clinicians and public health. We aim to provide a comprehensive analysis of evolving trends of antimicrobial resistance and ß-lactamases among E. coli and ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acine to bacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) in the Arabian region. A systematic review was conducted in Medline PubMed on papers published between January 2000 and February 2020 on countries in the Arab region showing different antibiotic resistance among E. coli and ESKAPE pathogens. A total of n = 119,144 clinical isolates were evaluated for antimicrobial resistance in 19 Arab countries. Among these clinical isolates, 74,039 belonged to E. coli and ESKAPE pathogen. Distribution of antibiotic resistance among E. coli and ESKAPE pathogens indicated that E. coli (n = 32,038) was the predominant pathogen followed by K. pneumoniae (n = 17,128), P. aeruginosa (n = 11,074), methicillin-resistant S. aureus (MRSA, n = 4370), A. baumannii (n = 3485) and Enterobacter spp. (n = 1574). There were no reports demonstrating Enterococcus faecium producing ß-lactamase. Analyses revealed 19 out of 22 countries reported occurrence of ESBL (Extended-Spectrum ß-Lactamase) producing E. coli and ESKAPE pathogens. The present study showed significantly increased resistance rates to various antimicrobial agents over the last 20 years; for instance, cephalosporin resistance increased from 37 to 89.5%, fluoroquinolones from 46.8 to 70.3%, aminoglycosides from 40.2 to 64.4%, mono-bactams from 30.6 to 73.6% and carbapenems from 30.5 to 64.4%. An average of 36.9% of the total isolates were reported to have ESBL phenotype during 2000 to 2020. Molecular analyses showed that among ESBLs and Class A and Class D ß-lactamases, blaCTX-M and blaOXA have higher prevalence rates of 57% and 52.7%, respectively. Among Class B ß-lactamases, few incidences of blaVIM 27.7% and blaNDM 26.3% were encountered in the Arab region. Conclusion: This review highlights a significant increase in resistance to various classes of antibiotics, including cephalosporins, ß-lactam and ß-lactamase inhibitor combinations, carbapenems, aminoglycosides and quinolones among E. coli and ESKAPE pathogens in the Arab region.

Molecular characterization of a multidrug-resistant/pandrug-resistant nosocomial polymicrobial infection with Klebsiella pneumoniae, Providencia rettgeri, and Acinetobacter baumannii from Rural Maharashtra, India.

The emergence of resistance against commonly used antibiotics has become a serious global concern. The rapid development of antibiotic resistance exhibited by Enterobacteriaceae has caused an increasing concern regarding untreatable bacterial infections. Here, we isolated four pathogens from a geriatric female patient who was hospitalized for a month with ventilator-associated pneumonia (VAP) and fever. The organisms isolated from the tracheal aspirates and urine included Klebsiella pneumoniae, pandrug-resistant Providencia rettgeri, and Acinetobacter baumannii. Resistome analysis indicated that the bacterial isolates from the polymicrobial infection were multiple-drug resitnat and pandrug resistant clones. Molecular characterization revealed presence of blaTEM-1 in K. pneumonaie, P. rettgeri and A. baumannii. The blaTEM-1 and blaNDM-1 genes were present in P. rettgeri and A. baumannii, whereas the blaTEM-1, blaNDM-1 and blaOXA-23 traits were present in A. baumannii isolates. The patient has died due to the unavailability of effective antimicrobial treatment for this drug-resistant polymicrobial infection.

Aerobic dichlorvos degradation by Pseudomonas stutzeri smk: complete pathway and implications for toxicity in Mus musculus.

BACKGROUND AND OBJECTIVES: Excess use of pesticides in agricultural field not only compromised soil fertility but also posed serious threat to water bodies and life in the surrounding environment. The leftover pesticide residue needs to be remediated effectively. Compared to physical, chemical and enzymatic remediation options the microbial remediation is more practical and sustainable. MATERIALS AND METHODS: The Pseudomonas stutzeri smk strain was found to use dichlorvos as the solitary carbon source. Minimal medium supplemented with dichlorvos was used to test ability of bacterium to degrade pesticide aerobically. The metabolites produced by the bacterium were studied with UV-Vis spectrophotometry, HPLC, FTIR and GC-MS techniques. The toxicity studies of neat dichlorvos and P. stutzeri smk degraded metabolites were studied by subcutaneous injection in Mus musculus. RESULTS: The P. stutzeri smk strain was found to degrade as high as 80% of dichlorvos on 7th day of incubation, at 30 °C temperature and at pH 7. In five steps complete aerobic degradation of 2,2dicholorvinyl dimethyl phosphate (dichlorvos) resulted in production of free methyl and phosphate. The degradation intermediates produced are 2-Chlorovinyl dimethyl phosphate, vinyl dimethyl phosphate, dimethyl phosphate, methylphosphate and finally free phosphate. The histopathological analysis of liver, spleen and thymus of M. musculus were performed to study toxicity of dichlorvos and degraded metabolites. CONCLUSION: P. stutzeri smk could result highest aerobic degradation of dichlorvos to produce free methyl and phosphate. Degradation metabolites could reverse largely toxic effects of dichlorvos when studied in M. musculus.

Unorthodox Parenteral ß-Lactam and ß-Lactamase Inhibitor Combinations: Flouting Antimicrobial Stewardship and Compromising Patient Care.

In India and China, indigenous drug manufacturers market arbitrarily combined parenteral ß-lactam and ß-lactamase inhibitors (BL-BLIs). In these fixed-dose combinations, sulbactam or tazobactam is indiscriminately combined with parenteral cephalosporins, with BLI doses kept in ratios similar to those for the approved BL-BLIs. Such combinations have been introduced into clinical practice without mandatory drug development studies involving pharmacokinetic/pharmacodynamic, safety, and efficacy assessments being undertaken. Such unorthodox combinations compromise clinical outcomes and also potentially contribute to resistance development.

Prevalence of ß-lactamase and antibiotic-resistant Pseudomonas aeruginosa in the Arab region.

BACKGROUND: Several studies in the Arab region have recognised the rate of nosocomial infections caused by Pseudomonas aeruginosa (P. aeruginosa), which produce ß-lactamase, and identified their emergence and prevalence in the region. This article reviewed molecular studies on these ß-lactamase-producing P. aeruginosa during 2010-2018 in several countries of the Arab region in order to analyse the trend of rising prevalence of disease causing drug-resistant P. aeruginosa in the Arab region. METHODS: Data from selected clinical studies during 2010-2018 on ß-lactamase-producing P. aeruginosa in the Arab region were obtained from reliable scientific databases for analysis and evaluation. RESULTS: Significant changes were found in resistance of P. aeruginosa towards certain antibiotics of the ß-lactam class. There was an increasing trend in the occurrence of resistance genes in ß-lactamase-producing P. aeruginosa. CONCLUSION: This review showed that there is an increasing prevalence of ß-lactamase-producing P. aeruginosa in some countries in the Arab region. This is a major cause of concern as this implies that more and more instances of multidrug resistance are emerging in this area. This leads to an overall negative impact on health concerns and amounts to increasing difficulty in combating disease. It is recommended that awareness about antibiotic use and abuse be made a priority and measures to curb unchecked use of prescription antibiotics be put into place. Effective screening methods to detect cases of resistance at their onset may be developed.

Preserving the Dwindling ß-lactams-Based Empiric Therapy Options for Gram-Negative Infections in Challenging Resistance Scenario: Lessons Learned and Way Forward.

Appropriate empiric therapy reduces mortality and morbidity associated with serious Gram-negative infections. ß-lactams (BLs) owing to their safety, efficacy, and coverage spectrum are the most preferred agents for empiric use. Inappropriate use of older penicillins and cephalosporins led to selection and spread of resistant clones. As a result, these valuable agents have lost their reliability compelling clinicians to often use erstwhile last-line therapies such as carbapenems. Excessive carbapenems use imposed collateral damage by selecting difficult-to-treat carbapenem-resistant organisms. Lack of empiric therapeutic options amenable for use in infections caused by contemporary pathogens was realized by the pharmaceutical industry leading to intensive efforts in discovering novel antibiotics. These efforts led to the approval of newer ß-lactams and ß-lactamase inhibitor (BL-BLI) combination. This review elaborates the past trends in empirical use of BLs and ensuing patterns of resistance emergence in Gram-negatives. Furthermore, a critical appraisal of newer BL-BLIs has been presented to identify the appropriate clinical situations for their use to ensure clinical efficacy coupled with minimal resistance selection. These learning have been derived from past trends of clinical usage of older empiric therapies so that the therapeutic utility of newer agents is preserved for long in light of dwindling global antibiotics pipeline.

The Calotropis Gigantea Methanolic Extract Induces Apoptosis in Human Breast Carcinoma Cells.

BACKGROUND: Calotropis gigantea (family: Asclepiadaceae) has been known to contain cardiac glycosides. The C. gigantea extracts have been reported as cytotoxic to a few cancer cell lines. The present study was designed to examine the effect of Calotropis gigantea methanolic extract (CGME) on the growth and apoptosis in human breast carcinoma cell line (MCF-7 cells). METHODS: The study was conducted in Aurangabad (India) from 16 February to 10 June 2015. CGME treated MCF-7 cells were analyzed for growth inhibition and apoptosis. The exhibition of phosphatidylserine was analyzed with the Annexin-V Fluorescein isothiocyanate flow cytometry (FITC) method. Accumulated poly-caspases were determined with carboxyfluorescein poly-caspase assay, Apo-BrdU™ tunnel assay for DNA fragmentation and pro/anti-apoptotic gene expression with real-time polymerase chain reaction. The high-performance liquid chromatography analysis indicated the presence of two unknown cardenolides along with known cardenolides such as calactin, calatropagenin, usharin, afroside, calatoxin, and gamphoside. The Kruskal-Wallis and Wilcoxon tests (GraphPad Prism version 7.0) were used for statistical analyses. RESULTS: Upon treatment with 40 µg/ml CGME, about 56.9% of the cell population underwent apoptosis. Compared to paclitaxel, the accumulation of active caspases in CGME treated with MCF-7 cells was found to be dose-dependent, whereas the G2/M cell cycle arrest was time-dependent. The Apo-BrdU™ tunnel assay confirmed that CGME treatment caused DNA fragmentation and RT-PCR analyses indicated elevated transcription for pro-apoptotic gene expression. Kruskal-Wallis test results were significant; Bcl-2 (P=0.00193), Bak-1 (P=0.00021), and Bax (P=0.0019). CONCLUSION: CGME treatment caused the accumulation of phosphatidylserine on the cell membrane, recruitment of poly-caspases, DNA fragmentation, and enhanced transcription of pro-apoptotic gene expression.

Data on isolation and purification of fibrinolytic enzyme from Pseudomonas baetica SUHU25.

The present dataset provides methodology to isolate and purify fibrinolytic enzyme from microbe isolated from the natural source. The information provided in this data article includes (1) isolation and identification of Pseudomonas baetica SUHU25, (2) optimization of cultural conditions, (3) extraction and purification of fibrinolytic enzyme, (4) protein estimation, (5) assay of fibrinolytic activity, (6) SDS PAGE for purified enzyme protein, (7) effect of pH, temperature and metal ions on fibrinolytic activity of enzyme protein, and (8) In-vitro blood clot dissolution assay.

Aerobic Degradation of Clothianidin to 2-Chloro-methyl Thiazole and Methyl 3-(Thiazole-yl) Methyl Guanidine Produced by Pseudomonas stutzeri smk.

Overuse of pesticides in agriculture may harm environmental and agricultural yields. Sustainable maintenance of soil fertility and management of the environment have become a concern due to the persistence of pesticides in the soil. Microbes have various mechanisms for the bioremediation of persistent organic pollutants from the environment. A bacterium that degrades clothianidin was isolated from the pesticide and applied to agricultural soil by the enrichment technique. The identity of the bacterium was determined by studying morphological, cultural, and biochemical characteristics and 16S rRNA gene sequences. The ability to metabolize clothianidin was confirmed using UV-visible spectrophotometric, chromatographic, and spectroscopic analyses. A Gram-negative bacterium, designated smk, isolated from clothianidin-contaminated soil was confirmed to be a member of Pseudomonas stutzeri. The biodegradation of clothianidin was studied using P. stutzeri smk. Approximately 62% degradation of clothianidin was achieved within two weeks when grown at 30°C and pH 7. The effects of various physicochemical parameters, including pH, temperature, and clothianidin concentrations, on catabolic rates were studied. The biodegradation studies using UV-Vis spectrophotometry, HPLC, FTIR, and LC-MS indicated the production of the following metabolites: 2-chloro-5-methyl thiazole (CMT), methyl nitroguanidine (MNG), methyl 3-[thiazole-yl], and methyl guanidine (TMG). Identification of specific degradation metabolites indicates that bioremediation of toxic neonicotinoid insecticides may be achieved by application of P. stutzeri smk.

Berberine from Argemone mexicana L exhibits a broadspectrum antibacterial activity.

The Argemone mexicana L, commonly found on desolate land in the Marathwada region of Maharashtra state, India, has been used for treating oral cavity infections. We sought to investigate the antimicrobial potential of A. mexicana L. In this study, cold aqueous and methanolic extracts were prepared from the A. mexicana L leaves. These extracts were tested for their antibacterial activities against selected bacterial isolates. The antibacterial activity and MICs were tested using the agar well diffusion method and broth dilution method, respectively. The cold aqueous and methanolic extracts of A. mexicana L leaves inhibited growth of clinical isolates of Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa.The antibacterial potentiality of A. mexicana L extracts was compared with Streptomycin - the reference antibiotic used in this study. The active ingredient of antibacterial potentiality within the A. mexicana L extract was purified and characterized by TLC, HPLC and NMR analysis. Structural elucidation of Berberine and its bioactivity both, from the A. mexicana L and commercial preparation, is investigated.