Role of Persistent Organic Pollutants in Breast Cancer Progression and Identification of Estrogen Receptor Alpha Inhibitors Using In-Silico Mining and Drug-Drug Interaction Network Approaches.

The strong association between POPs and breast cancer in humans has been suggested in various epidemiological studies. However, the interaction of POPs with the ERα protein of breast cancer, and identification of natural and synthetic compounds to inhibit this interaction, is mysterious yet. Consequently, the present study aimed to explore the interaction between POPs and ERα using the molecular operating environment (MOE) tool and to identify natural and synthetic compounds to inhibit this association through a cluster-based approach. To validate whether our approach could distinguish between active and inactive compounds, a virtual screen (VS) was performed using actives (627 compounds) as positive control and decoys (20,818 compounds) as a negative dataset obtained from DUD-E. Comparatively, short-chain chlorinated paraffins (SCCPs), hexabromocyclododecane (HBCD), and perfluorooctanesulfonyl fluoride (PFOSF) depicted strong interactions with the ERα protein based on the lowest-scoring values of -31.946, -18.916, -17.581 kcal/mol, respectively. Out of 7856 retrieved natural and synthetic compounds, sixty were selected on modularity bases and subsequently docked with ERα. Based on the lowest-scoring values, ZINC08441573, ZINC00664754, ZINC00702695, ZINC00627464, and ZINC08440501 (synthetic compounds), and capsaicin, flavopiridol tectorgenin, and ellagic acid (natural compounds) showed incredible interactions with the active sites of ERα, even more convening and resilient than standard breast cancer drugs Tamoxifen, Arimidex and Letrozole. Our findings confirm the role of POPs in breast cancer progression and suggest that natural and synthetic compounds with high binding affinity could be more efficient and appropriate candidates to treat breast cancer after validation through in vitro and in vivo studies.

Clonal relatedness and plasmid profiling of extensively drug-resistant New Delhi metallo-ß-lactamase-producing Klebsiella pneumoniae clinical isolates.

Aim: Carbapenem-resistant Klebsiella pneumoniae (CR-KP) particularly New Delhi metallo-ß-lactamase (NDM) is a serious public health concern globally. The aim of the study to determine the molecular epidemiology of blaNDM-producing clinically isolated K. pneumoniae. Methods: Carbapenem-resistant K. pneumoniae isolates (n = 100) were collected from tertiary care hospital Lahore. Isolates were confirmed by VITEK® 2 system and MALDI-TOF. Minimum inhibitory concentration was performed by VITEK 2 and molecular characterization was done by PCR, PFGE, DNA hybridization and replicon typing. Results: Of 90 MBL-producing K. pneumoniae, 75 were NDM producers; 60 were NDM-1 and 11 NDM-5. A total of 27 K. pneumoniae belonged to ST11 and 14 to ST147. NDM-positive isolates were 100% resistant to ß-lactam antibiotics except for colistin. 13.3% isolates carried blaNDM on ∼140 kb plasmids. A total of 32 (52.4%) isolates were positive for IncA/C and 18 (29.5%) IncF/II. Conclusion: The extensively resistant lineage of NDM-producing K. pneumoniae is prevalent in the clinical setting.

Polynuclear aromatic anthracene biodegradation by psychrophilic Sphingomonas sp., cultivated with tween-80.

Anthracene is a low molecular weight polynuclear aromatic hydrocarbons (PAHs) being identified as a precedence toxic contaminant in the ecosystem. Thus, the present work was designed to evaluate anthracene biodegradation efficiency by selected marine bacteria. From the marine isolates, the most effective anthracene biodegrading strain was identified as Sphingomonas sp., KSU05. Time course batch growth results indicated that the isolate KSU05 was capable of surviving up to 500 mg/L of anthracene. The influence of various nutrient sources were screened for enhanced growth and pyrene degradation, based on results glucose and tween-80 were used for further optimization studies. Batch experimental analysis showed maximum biodegradation (70.5%) of anthracene (50 mg/L) with enhanced survival of Sphingomonas sp. KSU05 was observed at 96 h of cultivation. Box-Behnken design optimization results showed that the culture conditions enhanced the anthracene biodegradation (90.0%) at pH 7.0, 0.3 mM of tween-80 concentration, and 5.5% of glucose concentration. In addition, the isolate Sphingomonas sp. KSU05 was found to rapidly degrade anthracene within 96 h. The anthracene intermediates was analyzed using Gas chromatography mass spectrophotometer (GC-MS). Overall, this research shown that the Sphingomonas sp., cultivated with suggested optimum conditions could provide an effective prospective for the degradation of anthracene from contaminated environment.

Molecular insights of Carbapenem resistance Klebsiella pneumoniae isolates with focus on multidrug resistance from clinical samples.

BACKGROUND: Carbapenem are the last-line antibiotic, defence against Gram-negative extended spectrum ß-lactamases producers (ESBLs). Carbapenem resistance Enterobacteriaceae especially Carbapenem resistant-Klebsiella pneumoniae (CR-KP) is recognized as one of the well-known public health problem, which is increasingly being reported around the world. The present study was focused to analyse the prevalence and characterization of antibiotic resistance K. pneumoniae in centre region of Tamil Nadu, India. METHODOLOGY: Totally 145 suspected K. pneumoniae isolates [Urine, Pus, Sputum, Blood and Biopsy] obtained from hospitals of Central South India. The isolates were subjected to biochemical and molecular identification technique, following with antibiotic resistance pattern by standard antibiotic sensitivity test. Multidrug resistance (MDR) with ß-lactamase producing Carbapenem resistant K. pneumoniae (CR-KP) strains were screened by classical sensitivity method and also drug resistance encoded gene. Also, molecular typing of the MDR strains were characterized by Pulsed-Field Gel Electrophoresis (PFGE). Further, the outer membrane protein (OmpK35 and 36) related Carbapenem resistance were characterized. RESULTS: Totally, 61% of isolates were confirmed as K. pneumoniae, 75 % of isolates were MDR including 58% carbapenem and 97% ESBL antibiotics and grouped into 17 distinct resistant patterns. The MDR KP isolates shows positive for blaCTXM-1 (92 %) gene followed by blaSHV (43 %), blaTEM (36 %), blaNDM-1 (26 %), blaGES (20 %) and blaIMP-1 (8 %). Moreover, 62 % CR-KP isolates loses OmpK36 and 33% isolates loses OmpK35. CONCLUSIONS: Loss of OmpK36 were highly an influence the cefoxitin and carbapenem resistance. Sixteen different PFGE patterns have been observed among the 18 MDR isolates. Eventually, ESBL as well as CR-KP were diverse in genetic makeup and often associated with hyper virulence hvKP should be of serious concern.

In vitro biological properties of Streptomyces cangkringensis isolated from the floral rhizosphere regions.

This context was investigated to determine in vitro antimicrobial, antioxidative, and anticancer traits of crude ethyl acetate extract of Streptomyces cangkringensis strain TSAS 04 isolated from soil sample of rhizosphere regions. The antimicrobial activity of ethyl acetate extract of strain TSAS 04 was determined against indicator pathogens using disc diffusion assay which exhibited maximum zones of inhibition of 20.6 ± 0.3 and 16.3 ± 0.6 mm against Bacillus subtilis and Trichoderma viride, respectively. In vitro antioxidant properties of the crude ethyl acetate extract were performed using standard methodologies. The extract revealed maximum DPPH and ABTS•+ radical scavenging activities of 51.1 ± 0.39 and 81.25 ± 0.33%, respectively. Likewise, maximum phosphomolybdenum reduction and Fe3+ reduction of the crude ethyl acetate extract of strain TSAS 04 were estimated 76.18 ± 0.10 and 89.01 ± 0.44%, respectively. In vitro anticancer trait of the extract was determined against HeLa cell line using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay which showed anticancer activities in a dose dependent manner with an IC50 value of 410.5 µg/mL. Fourier transform infrared spectroscopy (FT-IR) and Gas chromatography-mass spectrometry (GC-MS) analyses indicated the presence of distinct functional groups and bioactive components in the extract, respectively. In conclusion, S. cangkringensis strain TSAS 04 showed its effectiveness as ideal bioactive agent by exhibiting substantial antimicrobial, antioxidant, and anticancer properties.

Characterization of biofilm formed by multidrug resistant Pseudomonas aeruginosa DC-17 isolated from dental caries.

The present work reports with the screening of biofilm-producing bacteria from the dental caries. The dental pathogens showed resistance against various antibiotics and biofilm forming ability at various levels. Among the bacterial strain, Pseudomonas aeruginosa DC-17 showed enhanced biofilm production. Extracellular polymeric substance (EPS) was synthesized by the selected bacterial isolate considerably and contributed as the major component of biofilm. EPS composed of eDNA, proteins and lipids. The total protein content of the EPS was found to be 1.928 mg/mL and was the major component than carbohydrate and DNA. Carbohydrate content was 162.3 mg/L and DNA content of EPS was 4.95 µg/mL. These macromolecules interacted in the matrix to develop dynamic and specific interactions to signalling biofilm to differentiating various environments. Also, the isolated bacteria showed resistant against various commercially available antibiotics. The isolates showed more resistance against penicillin (98%) and were sensitive against amoxicillin. Among the factors, temperature, pH and sugar concentration influenced biofilm formation. Biofilm forming ability of the selected bacterial stain was tested at various pH values and alkaline pH was favoured for biofilm production. Biofilm production was found to be maximum at 40 °C and 8% sucrose enhanced biofilm formation. Biofilm formed by P. aeruginosa DC-17 was resistant against various tested antimicrobials and chemicals.

Exploring the potential of halotolerant bacteria for biodegradation of polycyclic aromatic hydrocarbon.

The present study aimed to determine the degradation and transformation of three-ring PAHs phenanthrene and anthracene by Cryptococcus sp. MR22 and Halomonas sp. BR04 under halophilic conditions. The growth progress of Cryptococcus sp. MR22 and Halomonas sp. BR04 on anthracene and phenanthrene was monitored by colony-forming unit (CFU) technique. The growth of the bacteria was maintained at a maximum concentration of 200 mg/L of all tested hydrocarbon, indicating that Cryptococcus sp. MR22 and Halomonas sp. BR04 significantly perform in the removal of the PAH-contaminated medium at low concentrations. The fit model to represent the biodegradation kinetics of both PAHs was first-order rate equation The extract prepared from cells supplemented with three different substrates exhibited some enzymes such as hydroxylase, dioxygenase, laccase and peroxidase. The results suggest that both strains had an impressive ability in the degradation of aromatic and aliphatic hydrocarbon but also could tolerate in the extreme salinity condition.

Anti-biofilm activity of plant derived extracts against infectious pathogen-Pseudomonas aeruginosa PAO1.

BACKGROUND: Biofilm forming ability of Pseudomonas aeruginosa make them vulnerable, because it makes them recalcitrant against various antibiotics. Quorum sensing (QS) is cell density based signaling that helps in bacterial cell-cell communication, which regulated various virulence factors such as pigment and biofilm formation that contribute in the establishment of chronic infections. The interruption of QS is one of the effective approach to control various virulence factors. Present study was intended with the aim to authenticate antibiofilm potential in different solvents based extracts of selected medicinal plant species viz. Berginia ciliata, Clematis grata and Clematis viticella traditionally used by the inhabitants of Himalayan region of Pakistan to treat various pathogenic diseases. P. aeruginosa PAO1, an opportunistic pathogen and involves in various life-threatening infections specifically in immune deficient patients was used as a model pathogen. METHODS: Plants were extracted in various organic (ethanol, methanol, acetone, ethyl acetate, hexane, chloroform) as well as in aqueous solvents and their ability to inhibit biofilm was measured. Biofilm of PAO1 was grown in Jensen's medium while growing at 30°C and crystal violet assay was performed to assess the biofilm inhibiting activity of plant extracts. RESULTS: Solvents play a vital role in extraction of plant components and it was found that the plants in various solvents exhibit different activity against the PAO1 biofilm. Comparatively, 1% methanolic extract of B. ciliata (rhizome with skin), showed more than 80% inhibition of biofilm formation without effecting on the growth of the bacterium. Significant correlation between flavonoids content and antibiofilm activity in methanolic extract revealed the contribution of secondary metabolites in P. aeruginosa (PAO1) biofilm inhibition. CONCLUSION: Our study revealed that plants under investigation more specifically B. ciliata could be a potential candidate for drug discovery to treat P. aeruginosa PAO1, induced infectious diseases especially for its biofilm treatment.

Molecular Detection of blaIMP Genes in Metallo-Beta-Lactamase Producing Clinical Gram-Negative Isolates.

BACKGROUND: The use of carbapenem antibiotics in the treatment of serious Gram-negative bacterial infections is under threat due to the emergence of the blaIMP gene amongst the bacterial pathogens. METHODS: The present descriptive cross-sectional study aimed to determine the occurrence of the blaIMP gene and minimum inhibitory concentrations (MICs) of the bacterial pathogens. The carbapenem-resistant isolates were screened out for the detection of MBLs by using the modified Hodge test and disk potentiation method. MBL producing strains were tested for the presence of the blaIMP gene by using PCR technique. The MICs of the blaIMP gene positive bacterial isolates were detected on the Vitek 2 system (bioMerieux). RESULTS: The primary source of MBLs and blaIPM gene carrying bacterial pathogens was blood (38.5%). We isolated 104 bacterial isolates in the initial screening of carbapenem resistance. Metallo-beta-lactamases (MBLs) were detected in 76 (73%) of the isolates which predominantly included 27 (26%) Klebsiella pneumoniae, 20 (19.2%) Acinetobacter baumannii, 16 (15.4%) Pseudomonas aeruginosa, and 11 (10.6%) E. coli while the other Gram-nega-tive MBL producing bacteria were few in number. The blaIPM gene was detected in 1 (1.3%) case of Acinetobacter baumannii and 1 (1.3%) case of Stenotrophomonas maltophilia. These strains were found to be multi-drug resistant with high MICs (≥ 8 to ≥ 256 µg/mL) against the majority of the drugs. CONCLUSIONS: The emergence of the blaIPM gene is a matter of serious concern as it left us with limited treatment options of minocycline, tigecycline, and levofloxacin. The horizontal transfer of blaIPM gene in other Gram-negative isolates can lead the epidemics of multidrug resistance.

Sonochemical synthesis and fabrication of honeycomb like zirconium dioxide with chitosan modified electrode for sensitive electrochemical determination of anti-tuberculosis (TB) drug.

Herein, novel honeycomb like zirconium dioxide with chitosan (ZrO2@chitosan) nanocomposite have been designed through a facile ultrasound-assisted method and followed by a simple sonication process (bath-type ultrasound washer; Honda Electronics-W-118T; 100 W/cm2 and 300 kHz frequency). After then, as-synthesized ZrO2@chitosan was characterized by FESEM, XRD and EIS. The ZrO2@chitosan nanocomposite modified glassy carbon electrode shows excellent electrochemical sensing performance towards anti-tuberculosis drug (rifampicin). Furthermore, the ZrO2@chitosan modified and fabricated electrochemical sensor showed a wide linear range between 0.015 µM and 547.4 µM and nanomolar detection limit (7.5 nM). Moreover, the ZrO2@chitosan modified electrode showed selectivity towards the detection of anti-tuberculosis drug (rifampicin). The ZrO2@chitosan nanocomposite film modified non-enzymatic sensor has high stable and good reproducible towards the detection of rifampicin. In addition, the as-synthesized ZrO2@chitosan nanocomposite modified electrode has been applied to the determination of rifampicin in biological samples such as human serum and urine samples.