Unveiling the role of hub proteins in controlling quorum sensing regulated virulence through analogues in Pseudomonas aeruginosa PAO1: A functional protein-protein network biology approach.

The protein-protein interaction (PPI) network analysis of specific genes identified for biofilm production and virulence/secretion system mediated by quorum sensing. The PPI depicted 13 hub proteins (namely rhlR, lasR, pscU, vfr, exsA, lasI, gacA, toxA, pilJ, pscC, fleQ, algR, and chpA) out of 160 nodes involving 627 edges. The PPI network analysis based on topographical features depicted pcrD with the highest degree value and vfr gene with the greatest betweenness centrality and closeness centrality (BC and CC) values. Based on in silico results, curcumin used as an Acyl homo-serine lactone (AHL) mimicker in P. aeruginosa, was also found effective in suppressing the quorum sensing regulated virulence factors such as elastase and pyocyanin. Based on in vitro experiment, curcumin suppressed biofilm formation at 62 µg/ml concentration. Host-pathogen interaction experiment showed that curcumin was also proved to be efficient in saving C. elegans from paralysis and killing effects of P. aeruginosa PAO1.

Functional analysis of Escherichia coli K12 toxin-antitoxin systems as novel drug targets using a network biology approach.

Bacterial resistance to various drugs and antibiotics has become a significant issue in the fight against infectious diseases. Due to the presence of diverse toxin-antitoxin (TA) systems, bacteria undergo adaptive metabolic alterations and can tolerate the effects of drugs and antibiotics. Bacterial TA systems are unique and can be therapeutic targets for developing new antimicrobial agents, owing to their ability to influence bacterial fate. With this background, our study aims to identify novel drug targets against Escherichia coli K12 MG1655 antitoxin using homology modelling approach. In this study, the protein-protein interaction network of 87 E. coli K12 MG1655 TA systems identified through literature mining was screened for the identification of hub proteins. The model evaluation, assessment, and homology modelling of the hub proteins were evaluated. Furthermore, computer-aided mathematical models of selected phytochemicals have been tested against the identified hub proteins. The TA system was functionally enriched in regulation of cell growth, negative regulation of cell growth, regulation of mRNA stability, mRNA catabolic process and RNA phosphodiester bond hydrolysis. RelE, RelB, MazE, MazF, MqsR, MqsA, and YoeB were identified as hub proteins. The robustness and superior quality of the RelB and MazE modelled structure were discovered by model evaluation, quality assessment criteria, and homology modelling of hub proteins. Clorobiocin was found to be a strong inhibitor by docking these modelled structures. Clorobiocin could be utilized as an antibacterial agent against multidrug resistant E. coli which may inactivate antitoxins and cause programmed cell death.

Microbial Sterility Testing for Mesenchymal Progenitor Cells Derived from Human Osteoarthritis and Rheumatoid Arthritis Cartilage.

BACKGROUND: The goal is to evaluate the microbial sterility of mesenchymal progenitor cells (MPCs)-derived from osteoarthritis (OA) and rheumatoid arthritis (RA) articular cartilages. METHODS: Contaminants, including bacteria and fungi in MPC cultures were initially evaluated by inoculation culture methods and then affirmed through the amplification of 16S ribosomal DNA (rDNA) and internally transcribed spacer (ITS) regions, respectively, using polymerase chain reaction (PCR). Further, the mollicutes, if any, were identified by genus-specific 16S rDNA, and the positive samples were reamplified using species-specific primers. RESULTS: No bacteria or fungi were found to be compromising the sterility of MPCs (n = 20) assessed by both traditional culture methods and PCR. However, two in early passages and three in later passages of MPCs had the presence of mollicutes. Further rescreening for species of mollicutes indicated the presence of Mycoplasma hyorhinis, M. salivarium, and M. arginine. CONCLUSIONS: The PCR methods employed in this study could be beneficial as a rapid sterility testing of cell lines.

Knowledge, Attitudes and HPV Vaccine Intention Among Women in India.

In India, cervical cancer associated with human papillomavirus (HPV) infection is a leading cause of cancer-related mortality among women. However, uptake of the vaccine in India is low. We assessed knowledge and attitudes towards HPV, assess participants' willingness to accept the vaccination for themselves and their children, and determine factors associated with intention to receive the HPV vaccine among women in Mangalore, India. This cross-sectional study surveyed a convenient sample of 237 women aged 18-45 years using a semi-structured questionnaire. All respondents reported being aware of HPV infection. However, 22.36% (n = 53) of the respondents have never heard about genital warts and 18.57% (n = 44) have never heard about HPV vaccine. Participants displayed good general knowledge of HPV infection (median score, 1.26; Interquartile Range (IQR): 1.04-1.52) and average knowledge of HPV vaccine (e.g., median score, 1.18; IQR: 0.73-1.45). HPV general knowledge and vaccine knowledge were associated with intention to receive the HPV vaccine and recommend it to children. Participant awareness of the HPV vaccine predicted vaccine intent for themselves. Participants' willingness to recommend the vaccine for their children was associated with older age, married status, having one or more children, and having a college education. Lack of awareness about genital warts was strongly associated with participants' refusal to get the HPV vaccine or recommend it for their own children (Relative Risk Ratio RRR: 12.21; 95% C.I.: 2.33-63.99). Our study validated the questionnaire as a reliable tool for assessing HPV and HPV vaccine knowledge, attitudes, awareness, and vaccine intentions in women aged 18-45 years. Public health education should focus on increasing awareness of genital warts as a sequela of HPV, as well as promote awareness of role and safety of HPV vaccination in -children.

In vitro Comparison of Adipogenic Differentiation in Human Adipose-Derived Stem Cells Cultured with Collagen Gel and Platelet-Rich Fibrin.

Background: Adipose-derived stem cells (ADSCs) are the most preferred cell type, based on their phenotypic characteristics, plasticity, and favorable immunological properties for applications in soft-tissue augmentation. Hence, the present in vitro study was aimed to evaluate the adipogenic differentiation potential of human ADSCs upon culturing individually with collagen gel and platelet-rich fibrin (PRF). Materials and methods: The collected lipoaspirate was used for establishing ADSCs using enzymatic digestion method. Then, the cells were analyzed for their morphology, viability, proliferation rate, population doubling time (PDT), colony-forming ability, cell surface markers expression, and osteogenic differentiation as biological properties. Further, ADSCs were evaluated for their adipogenicity using induction media alone, and by culturing with collagen gel and PRF individually for prospective tissue augmentation. Results: ADSCs were successfully established in vitro and exhibited a fibroblast-like morphology throughout the culture period. Cells had higher viability, proliferation potential and showed their ability to form colonies. The positive expression of cell surface markers and osteogenic ability confirmed the potency of ADSCs. The ADSCs cultured on collagen gel and PRF, individually, showed higher number of differentiated adipocytes than ADSCs grown with adipogenic induction medium alone. Conclusion: The extent of lipid accumulation by ADSCs was slightly higher when cultured on collagen gel than on PRF. Additional experiments are required to confirm better suitability of scaffold materials for soft-tissue regeneration.

Comparative Evaluation of Cytotoxicity and Genotoxicity of Two Bioceramic Sealers on Fibroblast Cell Line: An in vitro Study.

AIM: The purpose of this study was to evaluate and compare the cytotoxicity and genotoxicity of two bioceramic root canal sealers: EndoSequence BC and iRoot SP with zinc oxide eugenol sealers on fibroblast cell line. MATERIALS AND METHODS: The sealers tested were zinc oxide eugenol, EndoSequence BC, and iRoot SP. Each material was mixed according to the manufacturer's instructions and mounted into sterile polyethylene color-coded rings, for cytotoxicity and genotoxicity evaluation. After 48 hours, the set materials were transferred to previously marked wells and cytotoxicity evaluation to L929 murine fibroblast cells was done by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The percentages of viable cells were then calculated and values were statistically analyzed by Kruskal-Wallis test. The evaluation of genotoxicity of the materials to L929 murine fibroblast cells was carried out by Comet assay. To quantify deoxyribonucleic acid (DNA) damage, the following comet parameters were evaluated in the assay using Comet scoring software: tail length, tail moment, and Olive moment. The values were statistically analyzed using Kruskal-Wallis test with a significance value set to p < 0.05. RESULTS: The results of the study showed that both cytotoxicity and genotoxicity evaluation by MTT assay and Comet assay can be done on L929 murine fibroblast cell line. Among the three tested materials, zinc oxide eugenol showed maximum cytotoxicity to the cells (30.64% viable cells), followed by EndoSequence BC (71.33% viable cells) and iRoot SP (75.11% viable cells). The evaluation of DNA damage by genotoxicity assessment showed iRoot SP to be least genotoxic followed closely by EndoSequence BC. Zinc oxide eugenol was genotoxic and induced more DNA damage on the fibroblast cell line studied. The statistical analyses for both the assays were nonsignificant. CONCLUSION: All the three tested sealers showed varying degrees of cytotoxicity and genotoxicity while using fibro-blast cell line. Zinc oxide eugenol was most toxic in both the assays and iRoot SP showed least toxicity, followed closely by EndoSequence BC.

Knowledge and Attitudes Towards Human Papillomavirus (HPV) Among Academic and Community Physicians in Mangalore, India.

Cervical cancer is the most common cancer found in Indian women. Two human papillomavirus (HPV) vaccines were approved for use in India in 2006; however, neither has become readily accepted. Physician attitudes and recommendations are crucial in the uptake of HPV vaccines among adolescent women in the USA; thus, we ought to investigate provider attitudes and practices related to HPV vaccination in India via a survey administered to 210 Indian physicians. Of the 210 physicians, 46 % were community physicians and 54 % were academic physicians. The correct response to HPV knowledge questions was identified around 50 % of the time in 6/11 questions. Only 47 % of the physicians knew that there was an HPV vaccine approved for use in India. Only 11 % and 15 % of physicians strongly agree that the HPV vaccine will lead to long-lasting immunity and has a safe side effect profile, respectively. A total of 30 % of those surveyed reported that they would recommend the HPV vaccine to their patients, while 73 % agreed that the cost of the HPV vaccine is a major barrier to acceptance. After multivariate analysis, there were two significant variables independently associated with a physician's decision to recommend HPV vaccine. These variables were as follows: "whether the vaccine was freely available from the government sector" and "uncertainty about whether HPV must be persistent to cause cervical cancer vs not." Given the lack of knowledge among practicing physicians in Mangalore, increasing the education about HPV infection and HPV vaccination towards health care providers has the potential to increase vaccine recommendations.

Prevalence of Epstein Barr Virus and Herpes Simplex Virus Among Human Papillomavirus Negative Oral Cancer Patients: A Cross-Sectional Study from South India.

The high incidence of oral carcinomas is due to its multifactorial etiology and the presence of various risk factors. Human Papillomavirus (HPV) has a proven role in the pathogenesis of oral carcinomas, but in the recent times there has been an increasing incidence of oral cancers who are negative for HPV infection. Also, these patients are non-smokers and non-drinkers so it could be speculated that these oral cancers are due to some other etiological factor probably of other viral infections. Therefore, this study examined the prevalence of Epstein Barr Virus (EBV) and Herpes Simplex Virus (HSV) among oral cancer patients. This cross-sectional study was conducted from January 2019 to June 2020. Biopsy samples from 47 newly diagnosed untreated patients with oral malignancies were collected along with their demographic and clinicopathological information. DNA extracted from the biopsies was processed for nested PCR for the detection of EBV and HSV. All the samples tested negative for HPV and HSV infection. Nested PCR detected 29 cases (70.7%) to be positive for EBV. The non-cancerous adjacent tissues also were negative for HPV, EBV and HSV. The prevalence of EBV was found to be more in males (62.1%) and the highest number of cases was of the left buccal mucosa compromising 34% of the total cases. From the present study it can be concluded that EBV but not HSV infection is associated with an increased risk of developing oral cancers. Although, 70.7% of the patients were found to be positive for EBV whether the viral infection played any role in the driving the malignancy needs to be further elucidated.

Utilizing Bullseye fish processing frame waste to produce edible proteins and quality assessment of the recovered proteins.

The aim of the study was to utilize the waste generated from Bullseye (Priacanthus hamrur) fish processing to recuperate proteins. Considering the cost-effectiveness, versatility, and color improvement after the pH shift method, proteins from this waste were extracted by the pH shift method using hydrochloric acid and sodium hydroxide. The properties of extracted proteins were studied in detail. During the protein solubility measurement, maximum solubilization was found at pH 3.0 (13.10 mg/mL) on the acidic side and pH 11.0 (14.25 mg/mL) on the alkaline side with a total yield of 51.62 ± 0.23 and 45.42 ± 0.29 (%), respectively. The process variables tested in this study showed a significant effect on protein solubility (p < 0.05). The protein content of the isolates extracted from the waste was 23.80 ± 0.49 and 22.48 ± 0.39% for acid and alkali processed isolates, which was significantly higher than the mince (19.46 ± 0.67%). pH shift processing of Bullseye proteins caused a significant reduction in its pigments, lipids, and myoglobin content (p < 0.05). Proteins extracted using alkali had significantly higher values for foaming stability, water holding capacity, and emulsion capacity than proteins extracted using acid. An overall assessment indicated that protein isolates obtained using alkali extraction were better in terms of textural attributes, gelling ability, and amino acid profile than protein isolates extracted using the acid process.

Investigation of the hydration process and biological activity of a novel nanosilver incorporated dicalcium silicate based retrograde filling material.

Background: Although several materials have been used for retrograde filling following apical surgeries, there is no consensus on a single best material. Tricalcium silicate-based types of cement have been developed as root-end filling materials mainly due to tricalcium silicate's hydraulic properties. However, its unfavorable setting characteristics and minimal antimicrobial properties have necessitated the introduction of new additives into the existing commercially available materials. To design an affordable product based on a dicalcium silicate with a shorter set time, minimal cytotoxic complications, and enhanced antibacterial activity, we developed a new endodontic cement from pure raw materials, intending to satisfy the prerequisites of ideal retrograde material. Methods: The composition of the experimental calcium silicate-based cement included the addition of calcium chloride and silver nanoparticles in varying concentrations. Structural characterization was carried out using energy dispersive analysis by X-rays using scanning electron microscope (EDAX SEM) and hydration characteristics were performed using an X-ray diffractometer (XRD). The experimental material was further evaluated for biocompatibility using MTT ([3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide)assay and antibacterial activity was evaluated using an agar diffusion test against Enterococcus faecalis. Results: The structural characterization and hydration characteristics revealed that the experimental cement was dicalcium silicate based with favorable biocompatibility and enhanced antibacterial activity. Tricalcium silicate based mineral trioxide aggregate (MTA) also had favourable biocompatibility, however, its antibacterial activity was significantly decreased when compared to the novel cement. Conclusion: All hydraulic cements that are available in the dental market are predominantly tricalcium silicate-based materials. There has been no evidence in the literature to date wherein it has been explored whether a dicalcium silicate-based hydraulic cement can solely be used in root-end cavities. The findings of the study revealed a dicalcium silicate based retrograde filling material with favourable biocompatibility exhibited immediately as well as in the set samples. Incorporation of silver nanoparticles boosted the antibacterial activity when compared to that of ProRoot MTA. This material could potentially reinstate the usual hype created with tricalcium silicate types of cement since dicalcium silicate cements also exhibit similar properties.

Knowledge, Attitudes, Risk Perception, Preparedness and Vaccine Intent of Health Care Providers towards the Nipah Virus in South India.

Nipah virus (NiV) disease (NVD) remains a re-emerging public health threat in India. We assessed the knowledge, attitudes, and risk perception of NVD and future vaccine intent among a convenience sample of health care providers (HCP). The primary outcome measures were the knowledge, attitudes, and risk perception scores. Of 261 participants surveyed, 203 (77.8%) had heard of NiV and associated symptoms. The majority (248, 95%) identified the fruit bat as a primary NiV reservoir and 205 (79.8%) were aware of human-to-human transmission via droplets. Only 101 (38.7%) participants were aware that drinking date palm sap is a risk factor for transmission. Most HCP either agreed (117 (44.8%)) or strongly agreed (131 (50.2%)) that NiV is a serious illness. Less than half (121 (46.4%)) were aware of any institutional protocol for NiV; 235 (90.7%) of HCP stated that they need more information about prevention and treatment options. Knowledge scores were significantly higher among physicians compared to nurses whereas nurses and academic providers were more likely to have higher attitudes scores. A majority of respondents (20,779.9%) were willing to be vaccinated and willing to recommend the NiV vaccine to their patients (21,682.8%). Future strategies include education of HCP to bridge the knowledge gaps and enhance preparedness through disease-specific training for NiV infection.

A Comparative Evaluation of Antimicrobial Efficacy of Novel Surfactant-Based Endodontic Irrigant Regimen's on Enterococcus faecalis.

Introduction: Irrigants play an important role in the eradication of microorganisms in the complex root canal system. Sodium hypochlorite (NaOCl), chlorhexidine (CHX), and iodine potassium iodide (IKI) have certain limitations despite their routine clinical use. Surfactant irrigants with antimicrobial properties can be a milestone in endodontics to overcome the drawbacks of conventional irrigants. The aim of this study was to evaluate the antimicrobial efficacy of surfactants in comparison with routine endodontic irrigants on Enterococcus faecalis. Materials and Methods: Primary irrigants NaOCl, CHX, IKI were prepared at concentrations of 5%, 2.5%, 2%, and 1%; while mixture of tetracycline acid and detergent (MTAD) (100%) served as control group. Surfactants such as cetrimide (CTR) and sodium dodecyl sulfate (SDS) were prepared at concentrations of 0.5%, 1%, and 2%. The direct contact assay was used to evaluate the antimicrobial efficacy after 5 min. Statistical analysis was performed using one-way ANOVA and Mann-Whitney U-test. Results: In the primary group, only 5% CHX was significant over MTAD (P < 0.05). In the surfactant group, all concentrations of CTR and SDS (except 0.5%) were significant (P < 0.05) in killing E. faecalis over MTAD. Two percent CHX in combination with 0.5% CTR and 1% SDS had an effective kill percentage over 2% CHX and MTAD alone. Conclusion: Surfactant irrigant regimens can be used as adjuncts with CHX to overcome its clinical limitations and potentiate its substantivity, thereby enhancing clinical success in endodontics.

Assessment of proliferation, clonogenic assay, and osteogenic differentiation of human periodontal ligament stem cells following application of orthodontic forces.

Context: The proliferation and differentiation of human periodontal ligament stem cells (hPDLSC) into other cell types are also mediated by mechanical stresses; they might offer therapeutic benefits in tissue regeneration and angiogenesis. Objectives: The study was planned to assess the proliferation, clonogenic potential, and osteogenic differentiation of human periodontal ligament stem cells (PDLSC) following the application of light and heavy orthodontic forces. Materials and Methods: A couple forces of 50 gm (light force) were applied on the 1st premolar on the one side and 250 gm (heavy force) on the contralateral side in the upper arch of patients requiring orthodontic treatment with extraction of all 1st premolars. After 30 days, periodontal tissues were scrapped from extracted teeth for the establishment of PDLSC in vitro. PDLC from the lower premolar teeth where no orthodontic force was applied acted as the control group. Morphology, viability, proliferating rate and population doubling time, clonogenicity, and alkaline phosphatase activity were analysed. Result: The osteogenic potential was confirmed by Alizarin red staining and the expression of the osteogenic markers by qRT-PCR. The morphology, growth kinetics, potency, and osteogenic lineage characteristics inferred the application of high force reduced the proliferative ability and osteogenesis of PDLSC, though the difference was not significant. Conclusion: The established PDLSCs demonstrated their MSC-like properties based on morphology, growth kinetics, colony forming ability, and AP activity. The culture-expanded PDLSCs showed their differentiation potential into osteocytes. The application of high force reduced the proliferative ability and osteogenesis of PDLSCs, variations were not significant.

Comparative characterization and analysis of telomere length in stem cells derived from deciduous and permanent teeth.

Background: Understanding the influence of age on growth kinetics and telomere length in dental stem cells is essential for the successful development of cell therapies. Hence, the present study compared the basic cellular and phenotypical characteristics of stem cells from human exfoliated deciduous teeth (SHEDs) and dental pulp stem cells (DPSCs) of permanent teeth and their telomere lengths using quantitative real-time polymerase chain reaction. Materials and Methods: The study is an in vitro original research article. Primary cultures of SHED and DPSCs (n = 6 each) were successfully established in vitro, and the parameters analyzed were the morphology, viability, proliferation rate, population doubling time (PDT), phenotypic markers expression, and the relative telomere lengths. Data were analyzed by analysis of variance and P < 0.05 was considered statistically significant. Results: SHED and DPSCs exhibited a small spindle-shaped fibroblast-like morphology with >90% viability. The proliferation assay showed that the cells had a typical growth pattern. The PDT values of SHED and DPSCs were 29.03 ± 9.71 h and 32.05 ± 9.76 h, respectively. Both cells were positive for surface markers CD29, CD44, and CD90. However, they were negative for CD45 and human leukocyte antigen DR. Although the differences in relative telomere lengths between the individual cell lines of SHED and DPSCs were observed, no significant (P > 0.05) variations were found for the mean T/S ratios of both the cells. Conclusion: SHED and DPSCs displayed similar morphology, proliferation rates, and phenotypic features. The relative telomere lengths were slightly shorter in DPSCs than SHED, but the values were not significantly different. Thus, SHED and DPSCs can be considered as recognized sources for regenerative applications in dentistry.

Oncogenic viruses as etiological risk factors for head and neck cancers: An overview on prevalence, mechanism of infection and clinical relevance.

OBJECTIVE: This review article aims to discuss the role of oncogenic viruses in the development of head and neck cancers including the prevalence, mode of infection and the clinical relevance of these viral infections associated with tumours. DESIGN: A detailed review of scientific literature was performed relevant to oncogenic viruses associated with head and neck cancers. RESULTS: The incidence of head and neck cancers associated with traditional risk factors such as smoking, chewing tobacco and alcohol consumption have reduced gradually. With the emergence of oncogenic viruses, the viral infection has become a major etiological contributor to the global cancer burden. Viral infection in the etiology of cancer opens up an opportunity for viral gene specific targets in diagnosis and biomarkers to evaluate prognosis. Infection with high-risk HPVs in the oropharynx is already proving to be beneficial as a subset of HPV-positive oropharyngeal cancer patients tend to have better prognosis in terms of treatment responses and overall survival. CONCLUSIONS: Large multi-center clinical trials exploring the implications of modifying viral infections in cancers are further warranted and the results hold the key to the management of patients suffering from cancers driven by viral infections.

A full range of mouse sinoatrial node AP firing rates requires protein kinase A-dependent calcium signaling.

Recent perspectives on sinoatrial nodal cell (SANC)(*) function indicate that spontaneous sarcoplasmic reticulum (SR) Ca(2+) cycling, i.e. an intracellular "Ca(2+) clock," driven by cAMP-mediated, PKA-dependent phosphorylation, interacts with an ensemble of surface membrane electrogenic molecules ("surface membrane clock") to drive SANC normal automaticity. The role of AC-cAMP-PKA-Ca(2+) signaling cascade in mouse, the species most often utilized for genetic manipulations, however, has not been systematically tested. Here we show that Ca(2+) cycling proteins (e.g. RyR2, NCX1, and SERCA2) are abundantly expressed in mouse SAN and that spontaneous, rhythmic SR generated local Ca(2+) releases (LCRs) occur in skinned mouse SANC, clamped at constant physiologic [Ca(2+)]. Mouse SANC also exhibits a high basal level of phospholamban (PLB) phosphorylation at the PKA-dependent site, Serine16. Inhibition of intrinsic PKA activity or inhibition of PDE in SANC, respectively: reduces or increases PLB phosphorylation, and markedly prolongs or reduces the LCR period; and markedly reduces or accelerates SAN spontaneous firing rate. Additionally, the increase in AP firing rate by PKA-dependent phosphorylation by ß-adrenergic receptor (ß-AR) stimulation requires normal intracellular Ca(2+) cycling, because the ß-AR chronotropic effect is markedly blunted when SR Ca(2+) cycling is disrupted. Thus, AC-cAMP-PKA-Ca(2+) signaling cascade is a major mechanism of normal automaticity in mouse SANC.

Prevalence and Characterization of Extended-Spectrum ß-Lactamase-Producing Antibiotic-Resistant Escherichia coli and Klebsiella pneumoniae in Ready-to-Eat Street Foods.

As the global urban populations increase with rapid migration from rural areas, ready-to-eat (RTE) street foods are posing food safety challenges where street foods are prepared with less structured food safety guidelines in small and roadside outlets. The increased presence of extended-spectrum-ß-lactamase (ESBL) producing bacteria in street foods is a significant risk for human health because of its epidemiological significance. Escherichia coli and Klebsiella pneumoniae have become important and dangerous foodborne pathogens globally for their relevance to antibiotic resistance. The present study was undertaken to evaluate the potential burden of antibiotic-resistant E. coli and K. pneumoniae contaminating RTE street foods and to assess the microbiological quality of foods in a typical emerging and growing urban suburb of India where RTE street foods are rapidly establishing with public health implications. A total of 100 RTE food samples were collected of which, 22.88% were E. coli and 27.12% K. pneumoniae. The prevalence of ESBL-producing E. coli and K. pneumoniae was 25.42%, isolated mostly from chutneys, salads, paani puri, and chicken. Antimicrobial resistance was observed towards cefepime (72.9%), imipenem (55.9%), cefotaxime (52.5%), and meropenem (16.9%) with 86.44% of the isolates with MAR index above 0.22. Among ß-lactamase encoding genes, blaTEM (40.68%) was the most prevalent followed by blaCTX (32.20%) and blaSHV (10.17%). blaNDM gene was detected in 20.34% of the isolates. This study indicated that contaminated RTE street foods present health risks to consumers and there is a high potential of transferring multi-drug-resistant bacteria from foods to humans and from person to person as pathogens or as commensal residents of the human gut leading to challenges for subsequent therapeutic treatments.

Prevalence and Characterization of Methicillin-Resistant Staphylococcus aureus from Community- and Hospital-Associated Infections: A Tertiary Care Center Study.

The community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has become increasingly prevalent in both community and hospital settings. The aim of this study was to determine the prevalence, molecular characteristics and antibiotic resistance profiles of CA-MRSA from community- and hospital-associated infections in a tertiary care hospital in Mangalore, India. Of 520 S. aureus isolates, 362 were from inpatients (IP) and 158 were from outpatients (OP). One-hundred and thirty-two MRSA isolates obtained from 94 inpatients and 38 outpatients with complete clinical details were further analyzed. Of these, 81 (61.4%) were CA-MRSA (IP-47.9%, OP-94.7%) and 51 (38.6%) were HA-MRSA (IP-52.1%, OP-5.3%). All (100%) MRSA isolates were mecA gene positive. SCCmec typing identified SCCmec type IV (50.6%) and SCCmec type V (66.7%) in CA-MRSA, while SCCmec type I (41.2%), SCCmec type III (19.6%), SCCmec type IV (31.4%) and SCCmec type V (25.5%) were detected in HA-MRSA isolates. The Panton-Valentine Leukocidin (PVL) gene was found in 70.4% of CA-MRSA, 43.1% of HA-MRSA with SCCmec type IV and SCCmec type V, and in 7.8% of true HA-MRSA. The antibiotic resistance profiles were determined by the disc diffusion method. Resistance to cefoxitin was used to identify MRSA. A significant difference (p < 0.05) was observed between CA-MRSA and HA-MRSA with respect to resistance against cephalexin, cefotaxime, levofloxacin, linezolid and teicoplanin. CA-MRSA was predominantly resistant to ciprofloxacin (86.4%), erythromycin (66.7%), ofloxacin (49.4%), cefotaxime (44.4%), gentamicin (40.7%) and clindamycin (40.7%), while HA-MRSA showed resistance against ciprofloxacin (80.4%), erythromycin (80.1%), cefotaxime (70.6%),ofloxacin (58.8%), clindamycin (47.1%) and levofloxacin (41.2%).This study reports the prevalence of CA-MRSA in community and hospital settings and the possibility of multidrug-resistant CA-MRSA replacing HA-MRSA in hospitals. The observations from our study emphasize the need for urgent measures to manage this emerging crisis in healthcare settings.

Challenges to Human Papillomavirus Vaccine Acceptability among Women in South India: An Exploratory Study.

Cervical cancer is the second leading cause of cancer and one of the leading causes of cancer-related death in women in India. Human papillomavirus (HPV) vaccine uptake in India is low due to cost, low awareness of HPV, social stigma, and other factors. We assessed the awareness, attitudes, and beliefs regarding HPV and HPV vaccination and explored the barriers and challenges to HPV vaccine intent among women in Mangalore, India. An exploratory study was conducted using two focus group discussions (FGDs) and six in-depth one-on-one interviews. FGD-1 comprised nine women aged 18 to 26 years, and FGD-2 comprised seven women aged 27 to 45 years. The FGDs were recorded, transcribed, and analyzed using thematic content analysis. Themes identified were limited knowledge of HPV and vaccine, stigma associated with receiving HPV vaccine, vaccine safety concerns, and cost as a barrier to receiving vaccine. Participants expressed desire for physician and government recommendation of the HPV vaccine to validate vaccine intent. Contrasting themes between the two FGDs include support for vaccination at a younger age and lower perception of stigma and judgment in the 18- to 26-year-old group; however, participants in the 27- to 45-year-old group support vaccination at an older age and endorse greater fear of stigma and judgment associated with obtaining vaccination. Education regarding HPV-associated diseases and the HPV vaccine for the general public, physicians, and government officials in conjunction with lowering vaccine cost, improving vaccine access, and encouraging strong physician recommendations are key strategies to scale up HPV vaccine implementation in India.

Minimal influence of chronic inflammation on the potency and differentiation characteristics of gingiva-derived mesenchymal stem cells-An in vitro study.

OBJECTIVE: Gingiva-derived mesenchymal stem cells (GMSCs) have been identified and characterized from healthy tissues. However, reports on the influence of chronic inflammation on their stemness characteristics are sparse. The present study evaluated the potency and differentiation ability of GMSCs from periodontally healthy GMSC (H-GMSC) and inflamed GMSC (I-GMSC) tissues. MATERIALS AND METHODS: Established H-GMSCs and I-GMSCs were evaluated on their potency characteristics, such as morphology, viability, proliferation rate, population doubling time, colony-forming ability, expression of stemness markers, and mesenchymal differentiation potential. RESULTS: H-GMSCs and I-GMSCs exhibited fibroblast-like morphology and showed >95% viability with high proliferation potential and shorter doubling time. H-GMSCs showed fewer and smaller colonies, whereas I-GMSCs exhibited multiple and larger colonies. The evaluation of stemness markers revealed that both H-GMSCs and I-GMSCs were weakly positive for stage-specific embryonic antigen-4, Stro1, and CD105 (Endoglin), strongly positive for CD73 and CD90, and negative for the hematopoietic cell markers, CD34 and CD45. H-GMSCs showed a slightly higher osteogenic potential when compared to I-GMSCs, while I-GMSCs had a higher adipogenic potential than H-GMSCs. CONCLUSION: The findings showed that the inflammatory environment might have a stimulatory effect on the growth kinetics and ability of colony formation in GMSCs. However, varied osteogenic and adipogenic differentiation was observed between H-GMSCs and I-GMSCs.