Prevalence and Molecular Characterization of Metallo ß-Lactamase Producing Gram-Negative Pathogens Causing Eye Infections.

Purpose: Metallo ß-lactamases (MßL) production is a worldwide problem, particularly in gram-negative bacteria. As scanty data is available on the prevalence of MBL, the present study is being undertaken to determine the prevalence, antibacterial sensitivity patterns, and molecular characterization of MßL associated resistant genes in gram-negative bacteria isolated from ocular infections. Material and Methods: At a tertiary eye care center in south India, 359 gram-negative pathogens, 200 isolates from eye infections, and 159 isolates from normal flora of the eye were studied. A gold standard microbiology method was used to identify the isolates. An antibiotic double disc synergy test and a combination disc test were used to detect MßL production. Multiplex PCR was used to investigate the molecular characteristics of the MßL encoding genes blaVIM, blaIMP, and blaNDM. Results: Of the 359 gram-negative bacterial pathogens, Pseudomonas aeruginosa 108 (30.1%) and Enterobacter agglomerans 46 (12.8%) were commonly isolated. High prevalence of P. aeruginosa 81% (17 strains) was detected as an MßL producer and it shows 100% resistance to 2nd and 3rd generation cephalosporins and meropenem. Multiplex PCR detected only the blaVIM gene in 56 (28%) of various eye infections and 27 (17%) of normal flora of the gram-negative bacteria (GNB). The blaVIM gene is detected predominantly in 51.8% of keratitis and 21.4% of postoperative endophthalmitis. High prevalence of the gene was detected in P. aeruginosa 42.9% (24 of 56) and Alcaligens denitrificans 10.7% (6 of 56) from eye infections. Whereas, in the control group, P. aeruginosa and E. coli each had 14.8% (4 of 27) that were shown positive. Conclusion: The emerging MßLs mediated resistance among P. aeruginosa is a challenging task for ophthalmologists, especially in patients with endophthalmitis and bacterial keratitis. This local knowledge will aid in advising appropriate antibiotic use and avoiding unnecessary antibiotic prescriptions, which are highly warranted.

Potential Bioactive Compounds from Marine Streptomyces sp. and Their In Vitro Antibiofilm and Antibacterial Activities Against Antimicrobial-Resistant Clinical Pathogens.

Antimicrobial resistance issues have risen dramatically in recent years, posing a severe concern to humans worldwide. The urgent need to find novel compounds for pharmaceutical applications prompts the research of under-explored environments such as marine ecosystems. The present study was designed to discover novel secondary metabolites, and we have isolated about 30 actinomycetes from the marine soil samples collected in Thondi (Ramanathapuram, Tamil Nadu, India), where most isolates are associated with the genus Streptomyces. Out of 30, one potentially active strain (Streptomyces sp. SRMA3) was identified using primary and secondary screening methods against the drug-resistant clinical pathogens. The active metabolites extracted from the selected active isolate were subjected to partial purification and characterization using Fourier transform infrared spectrophotometer (FTIR) and gas chromatography-mass spectroscopy (GC-MS) analysis. The minimum inhibitory concentration (MIC) value was determined for the active metabolite. Further, the partially purified active fraction was revealed for its antibacterial and antibiofilm activity against drug-resistant clinical pathogens. Light and fluorescence microscopy detected the viability and adhesion of the biofilm-forming drug-resistant pathogens. Growth curve analysis showed that the active metabolite has the potential to inhibit drug-resistant pathogens. The synergistic effect of active metabolite with commercial antibiotics also revealed that it could enhance the activity of antibiotics in antimicrobial resistance pathogens. This study shows that the isolated Streptomyces sp. SRMA3 is a potentially active strain, and the metabolite derived from this strain has a good antibacterial and antibiofilm activity against antimicrobially resistant clinical pathogens and could be used for various biotechnological applications.

Phage in cancer treatment - Biology of therapeutic phage and screening of tumor targeting peptide.

INTRODUCTION: There is a constant drive to improve disease treatments. Much effort has been directed at identifying less immunogenic anti-cancer agents that produce fewer and less severe side effects. For more than a decade, bacteriophages have been discussed as an effective treatment for cancer with an exact mode of delivery. AREAS COVERED: We review how bacteriophages are used in cancer treatment, the underlying therapeutic mechanisms, and the tumor attacking peptide screening process. The filamentous bacteriophages are an effective vehicle for delivering displayed peptides toward the tumor target. The peptide must be expressed at the appropriate coat protein, and the peptide must be effective enough to disrupt the complex cancer matrix. The present review also sheds light on the dynamic use of phage in cancer treatment, from detection and diagnostics to treatment. EXPERT OPINION: Phage has a versatile role as a diagnostic and therapeutic tool. By acting as an appropriate recombinant drug, this phage has every potential to replace existing laborious, high capital investing therapies that may at many times result in failure or drastic side effects. One of the most significant challenges would be identifying tumor homing peptides. Although a few have been discovered, the most effective ones are yet to be determined. This therapeutic method plays a significant role in tumor therapy with high accuracy and efficiency, irrespective of the target location.

In Silico Identification of Hub Genes as Observing Biomarkers for Gastric Cancer Metastasis.

Perception of hub genes engaged in metastatic gastric cancer (mGC) promotes novel ways to diagnose and treat the illness. The goal of this investigation is to recognize the hub genes and reveal its molecular mechanism. In order to explore the potential facts for gastric cancer, the expression profiles of two different datasets were used (GSE161533 and GSE54129). The genes were confirmed to be part of the PPI network for gastric cancer pathogenesis and prognosis. In Cytoscape, the CytoHubba module was used to discover the hub genes. Responsible hub genes were identified. Data from Kaplan-Meier plotter confirmed the predictive value of these distinct genes in various stages of gastric malignancy. Upregulated and downregulated genes were identified to utilize for further analysis. Positive regulation by a host of viral process, positive regulation of granulocyte differentiation, negative regulation of histone H3-K9 methylation were found in DEGs analysis. In addition, five KEGG pathways were identified as an essential enhancer that include nucleotide excision repair; base excision repair; DNA replication; homologous recombination; and complement and coagulation cascades. POLE, BUB1B, POLD4, C3, BLM, CCT7, PRPF31, APEX1, PSMA7, and CDC45 were chosen as hub genes after combining the PPI results. Our study recommends that BUB1B, CCT7, APEX1, PSMA7, and CDC45 might be potential biomarkers for gastric cancer. These biomarkers are upregulated genes. Therefore, suppression of these genes will increase the survival rate in gastric cancer patients.

Medicinal plants mediated the green synthesis of silver nanoparticles and their biomedical applications.

The alarming effect of antibiotic resistance prompted the search for alternative medicine to resolve the microbial resistance conflict. Over the last two decades, scientists have become increasingly interested in metallic nanoparticles to discover their new dimensions. Green nano synthesis is a rapidly expanding field of interest in nanotechnology due to its feasibility, low toxicity, eco-friendly nature, and long-term viability. Some plants have long been used in medicine because they contain a variety of bioactive compounds. Silver has long been known for its antibacterial properties. Silver nanoparticles have taken a special place among other metal nanoparticles. Silver nanotechnology has a big impact on medical applications like bio-coating, novel antimicrobial agents, and drug delivery systems. This review aims to provide a comprehensive understanding of the pharmaceutical qualities of medicinal plants, as well as a convenient guideline for plant-based silver nanoparticles and their antimicrobial activity.

Isolation, Identification, Characterization, and Plasmid Profile of Urinary Tract Infectious Escherichia coli from Clinical Samples.

Objective: In recent times, urinary tract infection (UTI) is one of the most widely recognized bacterial diseases all over the planet. UTI influences individuals of any age and gender. The target of this study is to concentrate on the recurrence of uropathogens, the antimicrobial susceptibility pattern of the isolates, and the plasmid profile of people from the government clinics of Karaikudi. Methods: From July 2017 to December 2017, 100 urine tests were gathered and handled for the isolation of pathogenic microbes. In total, 89 isolates were found from the samples collected. Results: Escherichia coli was discovered as the most common bacterial isolate screened from the UTI-infected people, accounting for 28.09 percent of all isolates. E. coli was seen to be the highest prevalent bacterium for UTI in all age groups and demonstrated resistance to routinely used medications, especially cefpodoxime and novobiocin, which have been 100 percent resistant. The E. coli isolates screened were positive for beta-lactamase and film generation, and they have strong antimicrobial resistance. As a result, the E. coli strains with the highest prevalence of virulence determinants have become more resistant to many medications because they support the microorganism in overcoming the host's defense and colonizing or entering the urinary system. The amplified 16S rRNA product was analyzed, and phylogenetic relationships were determined. The presence of TEM (56 percent), CTX-M (64 percent), SHV (40 percent), and OXA (60 percent) was discovered. Among E. coli isolates, CTX-M was the most common extended spectrum-beta lactamase (ESBL). Multiplex PCR was also used to identify the existence of CTX-M subgroups in E. coli isolates. Conclusion: Finally, we urge that antibiotic selection should be predicated on the awareness of the specific prevalence and that novel antimicrobial medicines for urinary infections be developed to combat the overuse of antibiotics.

Biomarkers for Breast Adenocarcinoma Using In Silico Approaches.

This work elucidates the idea of finding probable critical genes linked to breast adenocarcinoma. In this study, the GEO database gene expression profile data set (GSE70951) was retrieved to look for genes that were expressed variably across breast adenocarcinoma samples and healthy tissue samples. The genes were confirmed to be part of the PPI network for breast cancer pathogenesis and prognosis. In Cytoscape, the CytoHubba module was used to discover the hub genes. For correlation analysis, the predictive biomarker of these hub genes, as well as GEPIA, was used. A total of 155 (85 upregulated genes and 70 downregulated genes) were identified. By integrating the PPI and CytoHubba data, the major key/hub genes were selected from the results. The KM plotter is employed to find the prognosis of those major pivot genes, and the outcome shows worse prognosis in breast adenocarcinoma patients. Further experimental validation will show the predicted expression levels of those hub genes. The overall result of our study gives the consequences for the identification of a critical gene to ease the molecular targeting therapy for breast adenocarcinoma. It could be used as a prognostic biomarker and could lead to therapy options for breast adenocarcinoma.

Bioengineered phytomolecules-capped silver nanoparticles using Carissa carandas leaf extract to embed on to urinary catheter to combat UTI pathogens.

Rising incidents of urinary tract infections (UTIs) among catheterized patients is a noteworthy problem in clinic due to their colonization of uropathogens on abiotic surfaces. Herein, we have examined the surface modification of urinary catheter by embedding with eco-friendly synthesized phytomolecules-capped silver nanoparticles (AgNPs) to prevent the invasion and colonization of uropathogens. The preliminary confirmation of AgNPs production in the reaction mixture was witnessed by the colour change and surface resonance plasmon (SRP) band at 410nm by UV-visible spectroscopy. The morphology, size, crystalline nature, and elemental composition of attained AgNPs were further confirmed by the transmission electron microscopy (TEM), selected area electron diffraction (SAED), X-ray diffraction (XRD) technique, Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The functional groups of AgNPs with stabilization/capped phytochemicals were detected by Fourier-transform infrared spectroscopy (FTIR). Further, antibiofilm activity of synthesized AgNPs against biofilm producers such as Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were determined by viability assays and micrographically. AgNPs coated and coating-free catheters performed to treat with bacterial pathogen to analyze the mat formation and disruption of biofilm formation. Synergistic effect of AgNPs with antibiotic reveals that it can enhance the activity of antibiotics, AgNPs coated catheter revealed that, it has potential antimicrobial activity and antibiofilm activity. In summary, C. carandas leaf extract mediated synthesized AgNPs will open a new avenue and a promising template to embed on urinary catheter to control clinical pathogens.

Fatty Acid Methyl Esters From the Coral-Associated Bacterium Pseudomonas aeruginosa Inhibit Virulence and Biofilm Phenotypes in Multidrug Resistant Staphylococcus aureus: An in vitro Approach.

In an attempt to study the antibacterial, antivirulence and antibiofilm potentials of bacteria residing the tissue and surface mucus layers of the pristine corals, we screened a total of 43 distinct bacterial morphotypes from the coral Favites sp. Among the isolates, Pseudomonas aeruginosa strain CBMGL12 with showed antibacterial, antivirulence and antibiofilm activity against multidrug resistant pathogenic strains of Staphylococcus aureus (reference strain: MTCC96; community-acquired methicillin resistant strain: CA-MRSA). Extracellular products (ECP) from the coral-associated bacterium P. aeruginosa were solvent extracted, fractionated by chromatographic techniques such as silica column and HPLC-UV with concomitant bioassays guiding the fractionation of metabolites. Identification of bioactive chemical moieties was performed by FT-IR analysis, GC-MS/MS equipped with NIST library, 1H and 13C NMR spectral studies. We report the differential production of extracellular and cell-associated virulence and biofilm phenotypes in multi-drug resistant strains of S. aureus, post-treatment with the ECP containing aromatic fatty acid methyl esters (FAME) such as methyl benzoate and methyl phenyl acetate produced by a coral-associated bacterium. In conclusion, this study has identified antibacterial, antibiofilm and antivirulent FAME from the coral-associated P. aeruginosa for its ability to attenuate virulence and biofilms phenotypes in multi-drug resistant pathogenic strains of S. aureus.

Molecular modelling of urease accessory interaction proteins of Helicobacter Pylori J 99 and predicting an interruption in interaction by Vigna radiata Defensins.

Helicobacter pylori is the major causative agent of Gastric carcinoma. Significance of the urease accessory interaction proteins are emphasized in colonization of human gastric mucosa and efficient infection of H. pylori. Here an attempt is made to explore the structure and properties of urease accessory interaction proteins from Helicobacter pylori J 99. The proteins chosen for the study are ureH, ureI, nikR, groL and flgS based on the interaction map available from STRING database. The above mentioned proteins do not have a comprehensive three dimensional structure. Hence the models were generated using PSI-BLAST (Position Specific Iterative-Blast) and MODELLER 9V8. Physicochemical characterization encompasses pI, EC, AI, II and GRAVY. Secondary structure was predicted using PSI-PRED. Functional characterization was done by SOSUI and DISULFIND Servers and refinement of structure was done using Ramachandran plot analysis. RMS-Z values were calculated using Q-MEAN Server and CHIMERA was used for molecular simulation studies. Plant defensins from Vigna radiata are successfully docked to the modeled structures and thus interaction could be possibly prevented. These results will pave way for further selective inhibition of H. pylori colonization and in vivo survival by employing plant defensins from Vigna radiata (VrD1 & VrD2). The work will prove that plant defensins provides anticancer relief too.