Volatile Constituents in Essential Oil from Leaves of Withania adpressa Coss. Ex Exhibit Potent Antioxidant and Antimicrobial Properties against Clinically-Relevant Pathogens.

Withania adpressa Coss. ex is a plant used in traditional medications. Antioxidant, antibacterial, and antifungal properties of the essential oil from leaves of Withania adpressa Coss ex. (EOW) were investigated. EOW was extracted using a Clevenger apparatus, and its volatile compounds were characterized by GC-MS. Antioxidant potency was determined using DPPH, FRAP, and TAC assays. Antibacterial effects were determined vs. Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, and Streptococcus pneumonia; while its antifungal efficacy was determined vs. Candida albicans, Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum using the disc diffusion and minimum inhibitory concentration bioassays. A chromatographic analysis showed that EOW contained eight phytochemical compounds constituting 99.14% of the total mass of oil. Caryophyllene (24.74%), Longifolene (21.37%), δ-Cadinene (19.08%), and Carene (14.86%) were predominant compounds in EOW. The concentrations required to inhibit 50% of free radical (IC50) values of antioxidant activities of EOW were 0.031 ± 0.006 mg/mL (DPPH), 0.011 ± 0.003 mg/mL (FRAP), and 846.25 ± 1.07 mg AAE/g (TAC). Inhibition zone diameters of EOW vs. bacteria were 18.11 ± 0.5 mm (E. coli), 17.10 ± 0.42 mm (S. aureus), 12.13 ± 0.31 mm (K. pneumoniae), and 11.09 ± 0.47 mm (S. pneumoniae), while MIC values were 51 ± 3, 47 ± 5, 46 ± 3 and 31 ± 1 µg/mL, respectively. Inhibition zone diameters of EOW vs. fungi were 31.32 ± 1.32, 29.00 ± 1.5, 27.63 ± 2.10, and 24.51 ± s1.07 mm for A. flavus, C. albicans, F. oxysporum, and A. niger, respectively. MIC values were 8.41 ± 0.40, 28.04 ± 0.26, 9.05 ± 0.76, and 22.26 ± 0.55 µg/mL, respectively. Importantly, the highest dose of EOW (1 mg/mL) showed negligible (~5%) cytotoxicity against MCF-12, a normal human epithelial cell line derived from the mammary gland, thus underscoring its wide safety and selectivity against tested microbes. To sum it up, EOW has exhibited promising antioxidant and antimicrobial properties, which suggests potential to abrogate antibiotic resistance.

Sericin-functionalized GNPs potentiate the synergistic effect of levofloxacin and balofloxacin against MDR bacteria.

A gradual expansion in resistant bacterial strains against commercially available antibacterial agents is the serious concern of the given research. It poses critical problem for public health. Thus, the demand for new antimicrobial agents has increased the interest in newer technologies and innovative approaches are required to advance the diagnosis and elimination of causative organisms. In this study, the potential role of technologies based on gold nanoparticles (GNPs) has been evaluated. GNPs were synthesized by using a cysteine protease, sericin whose reducing properties were exploited to bioengineer NPs (SrGNPs) where sericin with the help of thiol groups encapsulated over the surface of GNPs. Further, SrGNPs were bioconjugated with levofloxacin (Levo) and balofloxacin (Balo) to increase the efficacy of these drugs. Here, the antibacterial action of SrGNPs and their bioconjugated counterparts comprising Levo (Levo-SrGNPs), Balo (Balo-SrGNPs), and Levo/Balo (Levo-Balo-SrGNPs) were examined against normal and multi-drug resistant (MDR) strains of E. coli and S. aureus. The minimum inhibitory concentration (MIC) of these bioconjugates against said bacteria were found less than their pure counterparts. Further, the synergistic role of SrGNPs in combination with Levo and Balo was also explained using Chou-Talalay (C-T) method. The synthesis and bioconjugation of SrGNPs were confirmed by UV-visible spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM), and zeta-potential.

Gene expression patterns of COX-1, COX-2 and iNOS in H. Pylori infected histopathological conditions.

BACKGROUND: Research indicates that Helicobacter pylori can inflict severe histological damage through the modulation of host-related genes. The current study investigated the effect of H. pylori genotypes in the outcome of disease, and the expression of anti-apoptotic related genes, COX-1, COX-2, and iNOS genes in benign, pre-malignant, and malignant lesions of gastric carcinogenesis. MATERIALS AND METHODS: Tissue samples from H. pylori positive patients were graded based on the genotype of the infected H. pylori strain. Expression of COX-1, COX-2 and iNOS was assessed using a combination of real-time PCR and immunohistochemistry. RESULTS: Gene expression studies confirmed that COX-2 and iNOS expression was highly and selectively induced in epithelium with premalignant changes such as atrophic conditions, metaplasia and dysplasia, suggesting an important role of these genes in the sequence to gastric carcinoma of the intestinal type. Furthermore, the expression of COX-2 and iNOS was also dependent on the genotype of H. pylori and subjects with genotype-1 exhibited significantly higher expressions of COX-2 and iNOS compared to other genotypes. Comparison of the expression levels among infected and uninfected individuals demonstrated significant difference in the expression pattern of COX-2 gene whereas iNOS expression was found only in subjects infected H. pylori (p < 0.001). Immunohistochemical staining showed 1.5619 folds higher propensity of COX-2 and 3.2941 folds higher intensity of iNOS expression in subjects infected with H. pylori genotype 1. CONCLUSION: The up-regulation of COX-2 and iNOS was associated with the genotype of the H. pylori strain and the presence of certain genotype may greatly affect early events during carcinogenesis.

Purification and MIC analysis of antimicrobial proteins from Cucumis sativus L. seeds.

BACKGROUND: Cucumis sativus L. (cucumber), from the family Cucurbitaceae, is a therapeutic plant with various pharmacological benefits, broadly utilized as a part of complementary medicine (e.g., Unani, Ayurveda, Siddha, and Traditional Chinese). In light of past research discoveries, this plant had been chosen to consider its potential antibacterial action. METHODS: Extracts were purified by dialysis and ion exchange chromatography strategy and then assayed for antibacterial activity against four standard pathogenic bacterial strains known to cause foodborne infections and spoilage of food and herbal drugs. Antimicrobial peptides were extracted from seeds using a sodium phosphate citrate (pH 7.2) - CTAB cradle (pH 6.0). RESULTS: The highest protein concentration was seen with elute fractions 1 and 3 (370 mg/mL) compared with elute fractions 2 and 4 (340 mg/mL). Among the bacteria utilized, E. coli was clearly the most sensitive out of selected four strains. CONCLUSION: Our results suggest that Cucumis sativus L seeds extracts have significant potentials as new antimicrobial agents.

Chemically modified tetracyclines an emerging host modulator in chronic periodontitis patients: A randomized, double-blind, placebo-controlled, clinical trial.

Although periodontal diseases are caused by some of the specific pathogens, most of the tissue damage is caused by the host reaction to disease and not actually by the infections. Therefore, host modulatory therapy (HMT) has advanced benefit for the treatment of periodontitis, which works basically by reducing tissue destruction and regeneration in periodontium by altering the critical aspects of host response regulation and up regulating defensive regenerative responses. The present study was conducted with the goal to test an innovative therapeutic option using chemically modified tetracycline in patients affected with generalized, moderate and severe chronic periodontitis. We assumed that CMT might have the potential to provoke an assessable clinical result and pharmacologically impede the level inflammatory flow. CMT (incyclinide) treated group had significantly higher CAL (clinical attachment) values than Placebo Control suggesting an improved CAL in CMT treatment. Host modulation therapy width incyclinide can be as an adjunct to conventional nonsurgical therapies without antimicrobial resistance. Progress was noticed in the clinical parameters but not the serum CRP level in our study establishing the role of CMTs in controlling chronic periodontitis. Also CMT treatment indicates its role in anti-inflammatory process as it inhibited IL-12 and TNF alpha but IL-10 level was not affected. However, more randomized placebo-controlled clinical trials with large sample size are required in order to authenticate the usage of CMTs in chronic periodontitis treatment. Based on this understanding, exploration of the novel, low-cost synthetic inhibitors that can be used as potential therapeutic agents, has been tested.

The effect of immune modulation of Streptococcus constellatus SC10 strain upon Acinetobactor baumannii infection.

Acinetobacter baumannii is a tiny, plump and rod-shaped Gram-negative coccobacillus, notorious as an opportunistic pathogen, which is now being considered by CDC as a serious public health threat. Preventing colonization may be a better therapeutic choice using microbiota to protect against A. baumannii transmission. Human microbiota not only regulates immune homeostasis as well provides essential health benefits therefore discovering the interface due microbiota in host immune system will allow to recognise novel immuno- or microbe-based therapies. Streptococcus constellatus frequently isolated from oral cavity was selected for this study to check if its guards against A. baumannii by improving the immune response and the immune signalling pathway. In a mouse model we explored various parameters which include mouse body weight, internal organ weight, level of sIgA and different cytokine changes to evaluate its prophylactic effect against A. baumannii. Administration of S. constellatus SC10 was able to control the weight lose induced by A. bumannii infection in both the protected and treated groups. Particularly resistant against infection was more in treated group where the different cytokine following administration of SC10 strain remarkably controlled the infection and induced more sIgA production. The level of different interleukins IL-4, IL-10, IL-12 and IFN-γ was suppressed in treatment and protected group after exposure of S. constellatus. The excretion of sIgA following infection was many folds high in both group treated and protected groups Study suggests that SC10 L16 have potential immunomodulating effect which is able to regulate cytokines and sIgA response at the early phase of infection and modulate the further progress of clinical symptoms during infection.

Screening, Purification and Characterization of Anionic Antimicrobial Proteins from Foeniculum Vulgare.

Foeniculum vulgare Mill., commonly called fennel, is a medicinal plant belonging to the Umbelliferae (Apiaceae) family, and is used in traditional medicine. Antibacterial peptides were isolated using sodium phosphate citrate buffer and, for extraction, cetyltrimethyl ammonium bromide (CTAB) buffer with pH 6, have been employed and antimicrobial activity tested against four reference strains. The extracted protein was subjected to 3 kDa dialysis and separation was carried out by DEAE-ion exchange chromatography and further proteins were identified by 2D gel electrophoresis. The results of Foeniculum vulgare elutes obtained from DEAE-ion exchange chromatography were tested for antibacterial activity. Elute 3 shows the highest antibacterial activity on Pseudomonas aeruginosa with a diameter of a zone of inhibition of 16 mm and IC50 value 25.02 (mcg/mL). Based on the findings of the wide usage in treatment of various ailments and day-to-day life, Foeniculum vulgare seeds were used in the present research and have shown promising antibacterial effects, which requires further proteomic research to authenticate the role of the anticipated proteins.

Increasing Awareness of Hypertensive Patients About Their Blood Pressure Readings During Clinic Visits in a Tertiary Hospital in Riyadh.

Introduction Hypertension (HTN) is one of the most important cardiovascular risk factors. It is associated with significant complications, such as coronary artery disease, stroke, and chronic kidney disease. Awareness among hypertensive patients regarding their blood pressure (BP) is low in the Kingdom of Saudi Arabia. Aim This study aimed to evaluate the awareness of patients regarding their BP readings and to identify which aspects of HTN they needed to be informed about. Patients and methods A descriptive cross-sectional study was conducted among hypertensive patients attending outpatient clinics at King Fahad Medical City, Riyadh, Saudi Arabia. An electronic questionnaire was used by a trained physician to collect data from patients during telephone interviews. The information included socio-demographic data (i.e., age, gender, and education), family history, compliance with medications, and BP measurements. Patients were asked to answer questions to assess their awareness of their BP readings. Results Of the 475 hypertensive patients included in this study, 32.7% were aged between 56 and 65 years and 60.4% were female. The proportion of patients who had knowledge of their target BP (systolic: 120-129 mmHg; diastolic: 80-84 mmHg) was 74.4%. The significant independent predictors of increased knowledge about BP were a high level of education, regular measurement of BP, and having received education about a healthy lifestyle. The significant independent predictor of decreased knowledge about BP was having an acceptable or poor understanding of chronic BP. Conclusion Hypertensive patients visiting the outpatient clinic at King Fahad Hospital had a good understanding of their target BP readings. Educated patients who regularly measured their BP and who received education about a healthy lifestyle tended to exhibit a higher motivation to achieve their BP targets. More research is needed to gain more insights into the knowledge of hypertensive patients and into how they manage their BP to determine the factors that influence their knowledge.

Comparative Metabolic Characterization of Extraintestinal Pathogenic Escherichia coli Blood Isolates from Saudi Arabia.

Background: The prevalence of bloodstream infections caused by extraintestinal pathogenic Escherichia coli (ExPEC) has increased substantially. E. coli ST131 is one of the dominant ExPEC clones among E. coli bacteremia population. Metabolism can trigger the pathogenesis of some bacterial isolates, and here we evaluated and compared the metabolic traits of E. coli bacteremia isolates including ß-lactamase (BL)/extended-spectrum ß-lactamase (ESBL)-positive and ESBL-negative isolates and ST131 and non-ST131 isolates. Methods: The metabolic profiles of thirty E. coli isolates, obtained from blood samples for hospitalized individuals at a tertiary healthcare facility in Riyadh, were determined using HiMedia carbohydrate test strips. The difference in the utilization ability between isolate groups was then statistically assessed. Results: Our data found that non-BL/ESBL producers were of low metabolic capacity compared with ESBL-positive isolates although the difference remained insignificant. Higher levels of utilization for some carbohydrates, such as fructose and trehalose, were detected among ST131 isolates when compared with non-ST131, and ST131 was also significantly associated with metabolizing rhamnose. The mean bio-score of both isolate groups was insignificant. We showed no link between metabolism and antimicrobial susceptibility profiles among tested blood isolates. Conclusion: ST131 blood isolates were slightly higher in their carbohydrate utilization activity than non-ST131. More importantly, ST131 isolates were significantly capable of metabolizing rhamnose. Future research should focus on the factors that might drive the success of major ExPEC clones such as ST131.

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.

Broad-Spectrum Antimicrobial, Antioxidant, and Anticancer Studies of Leaf Extract of Simarouba glauca DC In Vitro.

The current study aimed to screen the preliminary phytochemicals in the leaf extract of the medicinal plant Simarouba glauca and to analyze its potential antimicrobial, antioxidant and anticancer properties. The phytochemical profile of the methanol extract was analyzed, and bioactive compounds were identified using chromatography, FTIR and GCMS. Antimicrobial activity and Minimum Inhibitory Concentration (MIC) were determined against 14 bacterial and 6 fungal strains. Moreover, the synergistic effect of a plant extract with commercially available antibiotics was also evaluated using the checkerboard method. The ethanolic and methanolic extracts showed exclusive activity against S. aureus and profound activity against E. coli and S. marcescens. Upon comparing breakpoints, methanolic extract demonstrated higher antimicrobial activity with a MIC value of 3.2 mg/mL against the test pathogens. Furthermore, the extracts demonstrated potential antioxidant activity; methanol extract had higher antioxidant potential compared to the ethanol extract. The major proactive bioactive compound with maximum antioxidant capacity was observed to be terpenoids. The methanol extract of S. glauca showed significant cytotoxicity against the MCF-7 breast cancer cell line with an IC50 value of 16.12 µg/mL. The overall results of our work provide significant evidence for the usage of methanolic extract of S. glauca as an efficient ethnomedicinal agent and a potential candidate for relieving many human ailments.

Phytochemical Analysis and Antioxidant, Antibacterial, and Antifungal Effects of Essential Oil of Black Caraway (Nigella sativa L.) Seeds against Drug-Resistant Clinically Pathogenic Microorganisms.

Nigella sativa (NS) is a plant that has long been utilized in traditional medicine as a treatment for certain diseases. The aim of this work was to valorize the essential oil (EO) of this species by phytochemical analysis and antimicrobial and antioxidant evaluation. EO was extracted by hydrodistillation from the seeds of Nigella sativa (EO-NS). Phytochemical content of EO-NS was evaluated by use of gas chromatography coupled to mass spectrometry (GC-MS/MS). Antioxidant ability was in vitro determined by use of three assays: 2.2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing power (FRAP), and total antioxidant capacity (TAC) relative to two synthetic antioxidants: BHT and quercetin. Antimicrobial effect was evaluated against four clinically important bacterial strains (Staphylococcus aureus, ATCC 6633; Escherichia coli, K12; Bacillus subtilis, DSM 6333; and Proteus mirabilis, ATCC 29906) and against four fungal strains (Candida albicans, ATCC 10231; Aspergillus niger, MTCC 282; Aspergillus flavus, MTCC 9606; and Fusarium oxysporum, MTCC 9913). Fifteen constituents that accounted for the majority of the mass of the EO-NS were identified and quantified by use of GC-MSMS. The main component was O-cymene (37.82%), followed by carvacrol (17.68%), α-pinene (10.09%), trans-sabinene hydrate (9.90%), and 4-terpineol (7.15%). EO-NS exhibited significant antioxidant activity with IC50, EC50, and total antioxidant capacity (TAC) of 0.017 ± 0.0002, 0.1196 ± 0.012, and 114.059 ± 0.97 mg EAA/g, respectively. Additionally, EO-NS exhibited promising antibacterial activity on all strains under investigation, especially on E. coli K12 resulting in inhibition diameter of 38.67 ± 0.58 mm and a minimum inhibitory concentration (MIC) of 1.34 ± 0.00 µg/mL. Also, EO-NS had significant antifungal efficacy, with a percentage of inhibition of 67.45 ± 2.31% and MIC of 2.69 ± 0.00 µg/mL against F. oxysporum, MTCC 9913 and with a diameter of inhibition 42 ± 0.00 mm and MIC of 0.67 ± 0.00 µg/mL against C. albicans. To minimize development of antibiotic-resistant bacteria, EO-NS can be utilized as a natural, alternative to synthetic antibiotics and antioxidants to treat free radicals implicated in microbial infection-related inflammatory reactions.

Comparative phenotypic characterization identifies few differences in the metabolic capacity between Escherichia coli ST131 subclones.

BACKGROUND: Extraintestinal pathogenic Escherichia coli (ExPEC) is responsible for causing many infections such as urinary tract infections (UTIs). The current dissemination of the multidrug resistant (MDR) ExPEC clone, Escherichia coli sequence type 131 (E. coli ST131), poses a real threat to public health worldwide. This study aimed to determine and compare the metabolic capacity of a collection of ExPEC isolates including ST131, non-ST131 and various ST131 subclones, and sought to assess the association between antimicrobial resistance and metabolic capacity of ST131 isolates. METHODS: The metabolic activity of forty urine E. coli isolates, collected from in-patients hospitalized at tertiary hospital in Riyadh, was tested using KB009 Hi carbohydrate kit, and then statistically analysed to assess the difference in the metabolic profiles between ST131 and non-ST131 isolates, and between ST131 subclones. RESULTS: The data of this study found almost similar metabolic profiles between ST131 and non-ST131, suggesting that ST131 is not a metabolically unique clone of ExPEC. There was also no link between antimicrobial susceptibility profiles and high metabolic capacity of ST131 isolates. Testing the biochemical activity of isolates belonging to ST131 subclones found higher activity of H30 subclone than non-H30 isolates, however it revealed few significant differences between these subclones. CONCLUSION: This study demonstrated no difference in the metabolism of ST131 and non-ST131, although it uncovered the presence of few significant differences in the metabolic capacity between ST131 subclones. Carrying out whole-genome based studies on ST131 and its main subclones is essential to elucidate the genetic factors responsible for the success of particular ST131 subclones.

Prevalence and molecular characteristics of sequence type 131 clone among clinical uropathogenic Escherichia coli isolates in Riyadh, Saudi Arabia.

BACKGROUND: The antimicrobial resistance of extraintestinal pathogenic Escherichia coli (ExPEC) has progressively been reported worldwide. This resistance has been ascribed to global dissemination of a single E. coli clone, namely E. coli sequence type 131 (E. coli ST131). The main goal of this study is to determine the prevalence and molecular traits of ST131 and its subclones among E. coli clinical urine isolates in Riyadh, Saudi Arabia. METHODS: Sixty E. coli urine isolates, of different extended spectrum ß-lactamase (ESBL) carriage, were involved in this study. Molecular characterization was carried out to determine the ST131 status, phylogenetic groups and virulence carriage of these isolates. ST131 isolates were further tested to evaluate the prevalence of different phylogenetic groups, subclones and virulence carriage. RESULTS: Group B2 was the most common phylogroup from which E. coli isolates derived. Overall, 37 of 60 (61.7%) isolates belonged to ST131 clones. Of these, 19 (31.7%) isolates were from the H30 subclone, including 10 (16.7%) H30 non-Rx and 9 (15%) H30Rx. The remaining 18 (30%) ST131 isolates belonged to other non H30 subclones. H30 subclone was significantly higher in the virulence carriage in comparison to non H30 ST131 subclones. CONCLUSION: This study reported the prevalence and traits of clinical E. coli ST131 main subclones in Saudi Arabia. It also demonstrated the high prevalence of E. coli ST131 locally, and found different virulence genotypes and antimicrobial resistance phenotypes among ST131 subclones. In the future, preforming whole genome sequence-based studies on ST131 and its subclones is crucial to elucidate factors that drive the success of these organisms.

In-vitro antibacterial, antioxidant potentials and cytotoxic activity of the leaves of Tridax procumbens.

The present study explored the phytochemicals, antibacterial, antioxidant and cytotoxic effect of Tridax procumbens leaves. The leaves were dried and extracted with various organic solvents. The leaves contained the phytochemicals such as alkaloids, carbohydrates, polyphenols and tannins respectively. Antimicrobial potentials of the extracts were determined by performing the disc diffusion techniques. Results revealed that different organic solvents extracts namely methanol, ethanol and ethyl acetate extracts documented comparatively good activity against the studied microbial strains. The methanol extract of leaves of T. procumbens showed combatively better antioxidant potential. The tested plant leaf extract showed high activity against human lung cancer cells than breast cancer cell lines. 250 µg/ml plants extract showed 84 ± 2.8% toxicity against human lung cancer cells.

Phyto-Mediated Synthesis of Porous Titanium Dioxide Nanoparticles From Withania somnifera Root Extract: Broad-Spectrum Attenuation of Biofilm and Cytotoxic Properties Against HepG2 Cell Lines.

There is grave necessity to counter the menace of drug-resistant biofilms of pathogens using nanomaterials. Moreover, we need to produce nanoparticles (NPs) using inexpensive clean biological approaches that demonstrate broad-spectrum inhibition of microbial biofilms and cytotoxicity against HepG2 cell lines. In the current research work, titanium dioxide (TiO2) NPs were fabricated through an environmentally friendly green process using the root extract of Withania somnifera as the stabilizing and reducing agent to examine its antibiofilm and anticancer potential. Further, X-ray diffraction (XRD), Fourier transform infrared (FTIR), scanning electron microscopy (SEM), transmission electron micrograph (TEM), energy-dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), thermogravimetric analysis (TGA), and Brunauer-Emmett-Teller (BET) techniques were used for determining the crystallinity, functional groups involved, shape, size, thermal behavior, surface area, and porosity measurement, respectively, of the synthesized TiO2 NPs. Antimicrobial potential of the TiO2 NPs was determined by evaluating the minimum inhibitory concentration (MIC) against Escherichia coli, Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, Listeria monocytogenes, Serratia marcescens, and Candida albicans. Furthermore, at levels below the MIC (0.5 × MIC), TiO2 NPs demonstrated significant inhibition of biofilm formation (43-71%) and mature biofilms (24-64%) in all test pathogens. Cell death due to enhanced reactive oxygen species (ROS) production could be responsible for the impaired biofilm production in TiO2 NP-treated pathogens. The synthesized NPs induced considerable reduction in the viability of HepG2 in vitro and could prove effective in controlling liver cancer. In summary, the green synthesized TiO2 NPs demonstrate multifarious biological properties and could be used as an anti-infective agent to treat biofilm-based infections and cancer.

Incidence and antimicrobial profile of extended-spectrum ß-lactamase producing gram-negative bacterial isolates: An in-vitro and statistical analysis.

OBJECTIVES: There is an extensive incidence of extended-spectrum beta-lactamases (ESBLs), principally in the hospital environment across the world. The present study was designed to discover the frequency of ESBL-production among the clinical isolates of Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. The study also focused on determining their liability to the selected antimicrobials. METHODS: Two hundred ten (210) clinical specimens were tested for the occurrence of ESBL using the double-disc synergy test. The molecular, physicochemical, absorption, distribution, metabolism, excretion, and toxicity were checked through an online server. RESULTS: Among the screened clinical isolates, E. coli (n=44), K. pneumonia (n=34) and P. aeruginosa (n=14) were ESBL markers. The ESBL producing isolates exhibited co-resistance to diverse categories of antibiotics. It was observed that all the ESBL-producing isolates were sensitive towards imipenem and faropenem with minimal proportion of resistance. CONCLUSION: The imipenem and faropenem can be recommended as the drugs of selection due to a lesser amount of resistance as compared to other antibiotics in this study.

Bio-inspired facile fabrication of silver nanoparticles from in vitro grown shoots of Tamarix nilotica: explication of its potential in impeding growth and biofilms of Listeria monocytogenes and assessment of wound healing ability.

Novel, safe, and effective antilisterial agents are required in order to prevent Listeria monocytogenes infections and maintain food safety. This study synthesized silver nanoparticles (AgNPs) from the shoot extract of in vitro-grown Tamarix nilotica (TN) and characterized them using X-ray diffraction, Fourier transform infrared spectroscopy, UV-visible spectroscopy, dynamic light scattering, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDXS), and transmission electron microscopy (TEM). We also assessed the antilisterial potential of the synthesized TN-AgNPs by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against two strains of L. monocytogenes and L. innocua. TN-AgNPs (2×MICs) showed a significant decrease in growth in all Listeria test strains. Release of cellular content and cell morphology analysis of TN-AgNP-treated bacterial cells demonstrated the mechanism of bactericidal activity of AgNPs. In addition, TN-AgNPs induced a significant decrease in swimming motility (62-71%), biofilm formation (57-64%), and preformed biofilms (48-58%) in all Listeria test strains at sub-inhibitory concentrations. Microtitre plate assay results for biofilm inhibition were confirmed by SEM and CLSM visualization of TN-AgNP-treated and TN-AgNP-untreated Listeria test strains. TN-AgNPs also showed wound-healing activity in MCF-7 cells by inhibiting cell migration in a scratch plate assay. TN-AgNP-induced enhanced reactive oxygen species generation in treated cells could be a plausible reason for the biofilm inhibitory activity of AgNPs. TN-AgNPs having antilisterial, antibiofilm, and wound-healing properties can be effectively used to prevent L. monocytogenes infections in the food industry and healthcare.

Chlortetracycline-Functionalized Silver Nanoparticles as a Colorimetric Probe for Aminoglycosides: Ultrasensitive Determination of Kanamycin and Streptomycin.

Aminoglycosides (AMGs) have been extensively used to treat infectious diseases caused by Gram-negative bacteria in livestock and humans. A selective and sensitive colorimetric probe for the determination of streptomycin and kanamycin was proposed based on chlortetracycline-coated silver nanoparticles (AgNPs-CTC) as the sensing element. Almost all of the tested aminoglycoside antibiotics can rapidly induce the aggregation of AgNPs, along with a color change from yellow to orange/red. The selective detection of aminoglycoside antibiotics, including tobramycin, streptomycin, amikacin, gentamicin, neomycin, and kanamycin, with other types of antibiotics, can be achieved by ultraviolet (UV) spectroscopy. This developed colorimetric assay has ability to detect various AMGs using in-depth surface plasmon resonance (SPR) studies. With this determination of streptomycin and kanamycin was achieved at the picomolar level (pM) by using a UV-visible spectrophotometer. Under aqueous conditions, the linear range of the colorimetric sensor for streptomycin and kanamycin was 1000-1,1000 and 120-480 pM, respectively. The corresponding limit of detection was 2000 pM and 120 pM, respectively. Thus, the validated dual colorimetric and ratiometric method can find various analytical applications for the ultrasensitive and rapid detection of AMG antibiotics in water samples.

Food color 'Azorubine' interferes with quorum sensing regulated functions and obliterates biofilm formed by food associated bacteria: An in vitro and in silico approach.

Quorum sensing (QS) plays a crucial role in different stages of biofilm development, virulence production, and subsequently to the growth of bacteria in food environments. Biofilm mediated spoilage of food is one of the ongoing challenge faced by the food industry worldwide as it incurs substantial economic losses and leads to various health issues. In the present investigation, we studied the interference of quorum sensing, its regulated virulence functions, and biofilm in food-associated bacteria by colorant azorubine. In vitro bioassays demonstrated significant inhibition of QS and its coordinated virulence functions in Chromobacterium violaceum 12472 (violacein) and Pseudomonas aeruginosa PAO1 (elastase, protease, pyocyanin, and alginate). Further, the decrease in the production EPS (49-63%) and swarming motility (61-83%) of the pathogens was also recorded at sub-MICs. Azorubine demonstrated broad-spectrum biofilm inhibitory potency (50-65%) against Chromobacterium violaceum, Pseudomonas aeruginosa, E. coli O157:H7, Serratia marcescens, and Listeria monocytogenes. ROS generation due to the interaction between bacteria and azorubine could be responsible for the biofilm inhibitory action of the food colorant. Findings of the in vitro studies were well supported by molecular docking and simulation analysis of azorubine and QS virulence proteins. Azorubine showed strong binding to PqsA as compared to other virulent proteins (LasR, Vfr, and QscR). Thus, it is concluded that azorubine is a promising candidate to ensure food safety by curbing the menace of bacterial QS and biofilm-based spoilage of food and reduce economic losses.