Gallic acid inhibits Staphylococcus aureus RecA protein functions: role in countering antibiotic resistance in bacteria.

AIM: Recombinase RecA and its homologs play a key role in homologous recombination DNA repair and revive stalled replication fork DNA synthesis. RecA plays an essential role in the evolution of antibiotic-resistant strains via stress-induced DNA repair mechanisms during the SOS response. Accordingly, RecA has become an attractive target to slow down antibiotic resistance rates and prevent mutations in pathogenic bacterial species. METHODS AND RESULTS: We employed RecA conserved activities: DNA binding, displacement loop formation, strand exchange, ATP hydrolysis, and LexA cleavage, to elucidate the inhibitory role of gallic acid on S. aureus RecA functions. Gallic acid inhibition of the SOS response by western blot analysis and its antibacterial activity were measured. The gallic acid inhibited all the canonical activities of S. aureus RecA protein. CONCLUSION: The natural phenolic compound gallic acid interferes with RecA protein DNA complex formation and inhibits activities such as displacement loop formation, strand exchange reaction, ATP hydrolysis, and coprotease activity of S. aureus. Additionally, gallic acid can obstruct ciprofloxacin-induced RecA expression and eventually confer the inhibitory role of gallic acid in the SOS survival mechanism in S. aureus.

Sex determination using mandibular ramus flexure in South Indian population - A retrospective study.

Sex determination or sex estimation from a single or fragment of bone is always difficult in the absence of other bones from the same individual. The current study was an attempt to estimate the sex of an individual from the posterior ramus of mandible or the mandibular ramus flexure. A retrospective study was conducted using orthopantomographs (OPGs) of 200 males and 200 females between the age group of 20 - 70 years. Each radiographic image was examined for the presence of a flexure or notching on the posterior border of the ramus in relation to occlusal plane as the method followed by Loth & Henneberg 1996.The study resulted in samples that were correctly classified as females 59.5% and males 57.5 %. The overall correct sex estimation was achieved in 58.5% of the cases. The predictive accuracy or assessment was higher for females compared to males. Consequently, the posterior ramus of mandible or mandibular ramus flexure can be considered as supplementary rather than a definitive means of sex determination. Hence, it is preferable to include as many parameters as possible to attain optimal accuracy.

Characterization of Staphylococcus aureus RecX protein: Molecular insights into negative regulation of RecA protein and implications in HR processes.

Homologous recombination (HR) is essential for genome stability and for maintaining genetic diversity. In eubacteria, RecA protein plays a key role during DNA repair, transcription, and HR. RecA is regulated at multiple levels, but majorly by RecX protein. Moreover, studies have shown RecX is a potent inhibitor of RecA and thus acts as an antirecombinase. Staphylococcus aureus is a major food-borne pathogen that causes skin, bone joint, and bloodstream infections. To date, RecX's role in S. aureus has remained enigmatic. Here, we show that S. aureus RecX (SaRecX) is expressed during exposure to DNA-damaging agents, and purified RecX protein directly interacts physically with RecA protein. The SaRecX is competent to bind with single-stranded DNA preferentially and double-stranded DNA feebly. Significantly, SaRecX impedes the RecA-driven displacement loop and inhibits formation of the strand exchange. Notably, SaRecX also abrogates adenosine triphosphate hydrolysis and abolishes the LexA coprotease activity. These findings highlight the role of the RecX protein as an antirecombinase during HR and play a pivotal role in regulation of RecA during the DNA transactions.

Comparative Evaluation of Postoperative Pain in Primary Teeth Obturated With Zinc Oxide Eugenol versus Metapex: A Randomized Clinical Trial.

The fundamental objective of pulp treatment is to preserve the integrity and health of oral tissues. Pulp necrosis or irreversible pulpitis is the key indication for performing pulpectomy in primary teeth. It can be performed as a single-visit or multi-visit procedure using suitable obturating material. Various obturating materials can be used like zinc-oxide eugenol or metapex. A controversy reflects in literature and among experts in regard to single-visit or multi-visit endodontics. Hence, the purpose of this randomized clinical trial is to assess the incidence of postoperative pain with zinc oxide eugenol and metapex as an obturating material in single-visit and multiple-visit pulpectomy. The study was conducted at M S Ramaiah Dental College and Hospital, Bangalore in one year duration from 2020 to 2021. Two groups of children, forty in each group, aged between four to eight years, were included in the study. Groups were divided to perform single-visit and multi-visit endodontics with zinc-oxide eugenol and metapex as obturating material. The sample size of 40 were considered and allocated to respective group by concealment allocation. The conventional pulpectomy procedure was carried out. The post operative pain was assessed by using the pain scale through the telephonic conversation with the patient within 24 hours of treatment. Proportions were compared using Chi-square test of significance and the "p" value of less than 0.05 indicates statistical significance. The results showed, no statistically significant difference in teeth obturated with zinc oxide eugenol and metapex in both single-visit and multi-visit pulpectomy. The comparison of zinc oxide eugenol and metapex in single-visit group and multi-visit group had a 'p' value of 0.9233 and 0.4233 respectively. There were no differences between single and multi-visit treatment protocols with respect to the incidence of post operative pain. Single visit pulpectomy can be performed with added advantages with either zinc oxide eugenol or metapex.

trans-ferulic acid attenuates hyperglycemia-induced oxidative stress and modulates glucose metabolism by activating AMPK signaling pathway in vitro.

Adenosine monophosphate-activated protein kinase (AMPK) is a potent metabolic regulator and an attractive target for antidiabetic activators. Here we report for the first that, trans-ferulic acid (TFA) is a potent dietary bioactive molecule of hydroxycinnamic acid derivative for the activation of AMPK with a maximum increase in phosphorylation (2.71/2.67 ± 0.10; p < .001 vs. high glucose [HG] control) in hyperglycemia-induced human liver cells (HepG2) and rat skeletal muscle cells (L6), where HG suppresses the AMPK pathway. It was also observed that TFA increased activation of AMPK in a dose- and time-dependent manner and also increased the phosphorylation of acetyl-CoA carboxylase (ACC), suggesting that it may promotes fatty acid oxidation; however, pretreatment with compound C reversed the effect. In addition, TFA reduced the level of intracellular reactive oxygen species (ROS) and nitric oxide (NO) induced by hyperglycemia and subsequently increased the level of glutathione. Interestingly, TFA also upregulated the glucose transporters, GLUT2 and GLUT4, and inhibited c-Jun N-terminal protein kinase (JNK1/2) by decreasing the phosphorylation level in tested cells under HG condition. Our studies provide critical insights into the mechanism of action of TFA as a potential natural activator of AMPK under hyperglycemia. PRACTICAL APPLICATIONS: Hydroxycinnamic acid derivatives possess various pharmacological properties and are found to be one of the most ubiquitous groups of plant metabolites in almost all dietary sources. However, the tissue-specific role and its mechanism under hyperglycemic condition remain largely unknown. The present study showed that TFA is a potent activator of AMPK under HG condition and it could be used as a therapeutic agent against hyperglycemia in type 2 diabetes.

Expression and Characterization of the Staphylococcus aureus RecA protein: A mapping of canonical functions.

Recombinases are responsible for homologous recombination (HR), proper genome maintenance, and accurate deoxyribonucleic acid (DNA) duplication. Moreover, HR plays a determining role in DNA transaction processes such as DNA replication, repair, recombination, and transcription. Staphylococcus aureus, an opportunistic pathogen, usually causes respiratory infections such as sinusitis, skin infections, and food poisoning. To date, the role of the RecA gene product in S. aureus remains obscure. In this study, we attempted to map the functional properties of the RecA protein. S. aureus expresses the recA gene product in vivo upon exposure to the DNA-damaging agents, ultraviolet radiation, and methyl methanesulfonate. The recombinant purified S. aureus RecA protein displayed strong single-stranded DNA affinity compared to feeble binding to double-stranded DNA. Interestingly, the RecA protein is capable of invasion and formed displacement loops and readily performed strand-exchange activities with an oligonucleotide-based substrate. Notably, the S. aureus RecA protein hydrolyzed the DNA-dependent adenosine triphosphate and cleaved LexA, showing the conserved function of coprotease. This study provides the functional characterization of the S. aureus RecA protein and sheds light on the canonical processes of homologous recombination, which are conserved in the gram-positive foodborne pathogen S. aureus.

Heterotic pools in African and Asian origin populations of pearl millet [Pennisetum glaucum (L.) R. Br.].

Forty-five African or Asian origin pearl millet populations bred either in Africa or Asia were investigated to generate information on heterotic pools. They were clustered into seven groups (G1 to G7) when genotyped, using 29 highly polymorphic SSRs. Fourteen parental populations representing these seven marker-based groups were crossed in diallel mating design to generate 91 population hybrids. The hybrids evaluated at three locations in India showed mean panmictic mid-parent heterosis (PMPH) and better-parent heterosis (PBPH) for grain yield ranging from - 21.7 to 62.08% and - 32.51 to 42.99%, respectively. Higher grain yield and heterosis were observed in G2 × G6 (2462 kg ha-1, 43.2%) and G2 × G5 (2455 kg ha-1, 42.8%) marker group crosses compared to the most popular Indian open-pollinated variety (OPV) ICTP 8203. Two heterotic groups, Pearl millet Population Heterotic Pool-1 (PMPHP-1) comprising G2 populations and Pearl millet Population Heterotic Pool-2 (PMPHP-2) comprising G5 and G6 populations, were identified based on hybrid performance, heterosis and combining ability among marker group crosses. Population hybrids from two heterotic groups, PMPHP-1 × PMPHP-2 demonstrated PMPH of 14.75% and PBPH of 6.8%. Populations of PMPHP-1 had linkages with either African or Asian origin populations, whereas PMPHP-2 composed of populations originating in Africa and later bred for Asian environments. Results indicated that parental populations from the two opposite heterotic groups can be used as base populations to derive superior inbred lines to develop high yielding hybrids/cultivars.

Identification of heterotic groups in South-Asian-bred hybrid parents of pearl millet.

KEY MESSAGE: Pearl millet breeding programs can use this heterotic group information on seed and restorer parents to generate new series of pearl millet hybrids having higher yields than the existing hybrids. Five hundred and eighty hybrid parents, 320 R- and 260 B-lines, derived from 6 pearl millet breeding programs in India, genotyped following RAD-GBS (about 0.9 million SNPs) clustered into 12 R- and 7 B-line groups. With few exceptions, hybrid parents of all the breeding programs were found distributed across all the marker-based groups suggesting good diversity in these programs. Three hundred and twenty hybrids generated using 37 (22 R and 15 B) representative parents, evaluated for grain yield at four locations in India, showed significant differences in yield, heterosis, and combining ability. Across all the hybrids, mean mid- and better-parent heterosis for grain yield was 84.0% and 60.5%, respectively. Groups G12 B × G12 R and G10 B × G12 R had highest heterosis of about 10% over best check hybrid Pioneer 86M86. The parents involved in heterotic hybrids were mainly from the groups G4R, G10B, G12B, G12R, and G13B. Based on the heterotic performance and combining ability of groups, 2 B-line (HGB-1 and HGB-2) and 2 R-line (HGR-1 and HGR-2) heterotic groups were identified. Hybrids from HGB-1 × HGR-1 and HGB-2 × HGR-1 showed grain yield heterosis of 10.6 and 9.3%, respectively, over best hybrid check. Results indicated that parental groups can be formed first by molecular markers, which may not predict the best hybrid combination, but it can reveal a practical value of assigning existing and new hybrid pearl millet parental lines into heterotic groups to develop high-yielding hybrids from the different heterotic groups.

Inhibition of the pro-inflammatory mediators in rat neutrophils by shanzhiside methyl ester and its acetyl derivative isolated from Barleria prionitis.

ETHNOPHARMACOLOGICAL RELEVANCE: The aerial parts of Barleria prionitis Linn. (BP) (Acanthaceae) plant has long been used to treat inflammatory disorders such as toothache, swellings, arthritis and gout. AIM OF THE STUDY: The purpose of this study was to evaluate the effects of shanzhiside methyl ester (SME), 8-O-acetyl shanzhiside methyl ester (ASME) and iridoid glycosides rich monoterpenoidal fraction (IFBp), isolated from the aerial part of BP, on the pro-inflammatory mediators in stimulated rat neutrophils. MATERIALS AND METHODS: Rat neutrophils were incubated with or without test drugs. The influence of laboratory isolated and identified SME, ASME and IFBp on the production and release of pro-inflammatory mediators i.e. myeloperoxidase (MPO), elastase, matrix metalloproteinase-9 (MMP-9), interleukin 8 (IL-8), tumor necrosis factor alpha (TNF-α) and leukotriene B4 (LTB4) was evaluated in the formyl-met-leu-phenylalanine (f-MLP) and lipopolysaccharide (LPS) stimulated rat neutrophils using enzyme-linked immunosorbent assay (ELISA) methods. IFBp was also standardized with the high performance thin layer chromatography by simultaneous determination of SME and ASME marker compounds. RESULTS: SME, ASME and IFBp displayed concentration-dependent inhibitory effects on the MPO, elastase and MMP-9 enzymes release, and IL-8, TNF-α and LTB4 cytokines production in the f-MLP and LPS stimulated rat neutrophils. The content of SME and ASME was found to be 17.32 ±â€¯1.98 and 11.30 ±â€¯1.06% w/w, respectively, in IFBp by HPTLC method. CONCLUSION: Altogether, the present results suggest that the iridoidal glycosides of BP may be considered as therapeutic strategy against neutrophil-mediated inflammatory diseases. Developed and validated HPTLC method for the standardization of IFBp of BP can be used as a quality control tool for the routine qualitative and quantitative analysis of Barleria species containing SME and/or ASME.

Molecular and functional characterization of the Listeria monocytogenes RecA protein: Insights into the homologous recombination process.

The recombinases present in the all kingdoms in nature play a crucial role in DNA metabolism processes such as replication, repair, recombination and transcription. However, till date, the role of RecA in the deadly foodborne pathogen Listeria monocytogenes remains unknown. In this study, the authors show that L. monocytogenes expresses recA more than two-fold in vivo upon exposure to the DNA damaging agents, methyl methanesulfonate and ultraviolet radiation. The purified L. monocytogenes RecA protein show robust binding to single stranded DNA. The RecA is capable of forming displacement loop and hydrolyzes ATP, whereas the mutant LmRecAK70A fails to hydrolyze ATP, showing conserved walker A and B motifs. Interestingly, L. monocytogenes RecA and LmRecAK70A perform the DNA strand transfer activity, which is the hallmark feature of RecA protein with an oligonucleotide-based substrate. Notably, L. monocytogenes RecA readily cleaves L. monocytogenes LexA, the SOS regulon and protects the presynaptic filament from the exonuclease I activity. Altogether, this study provides the first detailed characterization of L. monocytogenes RecA and presents important insights into the process of homologous recombination in the gram-positive foodborne bacteria L. monocytogenes.

p-Coumaric acid inhibits the Listeria monocytogenes RecA protein functions and SOS response: An antimicrobial target.

Bacterial RecA plays an important role in the evaluation of antibiotic resistance via stress-induced DNA repair mechanism; SOS response. Accordingly, RecA became an important therapeutic target against antimicrobial resistance. Small molecule inhibitors of RecA may prevent adaptation of antibiotic resistance mutations and the emergence of antimicrobial resistance. In our study, we observed that phenolic compound p-Coumaric acid as potent RecA inhibitor. It inhibited RecA driven biochemical activities in vitro such as ssDNA binding, strand exchange, ATP hydrolysis and RecA coprotease activity of E. coli and L. monocytogenes RecA proteins. The mechanism underlying such inhibitory action of p-Coumaric acid involves its ability to interfere with the DNA binding domain of RecA protein. p-Coumaric acid also potentiates the activity of ciprofloxacin by inhibiting drastic cell survival of L. monocytogenes as well as filamentation process; the bacteria defensive mechanism in response to DNA damage. Additionally, it also blocked the ciprofloxacin induced RecA expression leading to suppression of SOS response in L. monocytogenes. These findings revealed that p-Coumaric acid is a potent RecA inhibitor, and can be used as an adjuvant to the existing antibiotics which not only enhance the shelf-life but also slow down the emergence of antibiotic resistance in bacteria.

Eficacia del láser de diodo de arseniuro de galio en el tratamiento de 2 casos de liquen plano oral / Effectiveness of 904 nm Gallium-Arsenide Diode Laser in Treatment of Oral Lichen Planus: Report of 2 Cases

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Expression, purification and biochemical characterization of Listeria monocytogenes single stranded DNA binding protein 1.

Single-stranded DNA binding proteins play an important role in DNA metabolic processes including replication, recombination, and repair. Here, we report the identification and biochemical characterization of the SSB1 protein from the foodborne pathogen Listeria monocytogenes. The L. monocytogenes SSB1 share 33% identity and 50.5% similarity with the prototype E. coli SSB protein. The electrophoretic mobility shift assay revealed that the purified L. monocytogenes SSB1 protein binds to single stranded DNA, including the M13 circular single stranded DNA and oligonucleotide, with high affinity. The plasmid based strand transfer activity showed that, in the absence of the SSB protein, L. monocytogenes RecA fails to catalyze the reaction whereas, the E. coli RecA protein has shown nicked DNA formation. Interestingly the addition of SSB1 protein stimulates both L. monocytogenes and E. coli RecA strand transfer activities however, it is sensitive to the order of addition of SSB1 protein. L. monocytogenes RecA fails to catalyze the reaction when SSB1 is added prior to RecA; nevertheless, it readily catalyzes the reaction when added after the RecA filament formation. These results suggest that the interaction among of gene product between RecA and SSB1 is required to promote optimum strand exchange activities. Altogether, these studies provide the first functional characterization of the L. monocytogenes SSB1 protein and give insights into DNA repair and recombination processes in the gram-positive foodborne pathogen L. monocytogenes.

Two sample comparisons including zero-inflated continuous data: A parametric approach with applications to microarray experiment.

Micro-array experiments are important fields in molecular biology where zero values mixed with a continuous outcome are frequently encountered leading to a mixed distribution with a clump at zero. Comparison of two mixed populations, e.g. of a control and a treated group; of two groups with different types of cancer, to name a few, are often encountered in these contexts. Fairly skewed distribution of the continuous part coupled with small sample sizes are issues of main concern to be attended for the quality of inference in such situations while popularly used nonparametric methods rely on asymptotic distribution of the underlying test statistics which are valid only under large sample sizes. We address the aforementioned issues via a newly proposed exact test for location-scale family distributions and Generalized pivot quantity (GPQ) based parametric test procedures for non-location-scale distributions. Simulation based assessment showed their superior performance with respect to size and power in comparison to the popular two-part tests (Wilcoxon rank sum, t-test, Kolmogrov-Smirnov, Ansari-Bradley and Sigel-Tukey) more prominently for small sample sizes.

Identification of mega-environments and rice genotypes for general and specific adaptation to saline and alkaline stresses in India.

In the present study, a total of 53 promising salt-tolerant genotypes were tested across 18 salt-affected diverse locations for three years. An attempt was made to identify ideal test locations and mega-environments using GGE biplot analysis. The CSSRI sodic environment was the most discriminating location in individual years as well as over the years and could be used to screen out unstable and salt-sensitive genotypes. Genotypes CSR36, CSR-2K-219, and CSR-2K-262 were found ideal across years. Overall, Genotypes CSR-2K-219, CSR-2K-262, and CSR-2K-242 were found superior and stable among all genotypes with higher mean yields. Different sets of genotypes emerged as winners in saline soils but not in sodic soils; however, Genotype CSR-2K-262 was the only genotype that was best under both saline and alkaline environments over the years. The lack of repeatable associations among locations and repeatable mega-environment groupings indicated the complexity of soil salinity. Hence, a multi-location and multi-year evaluation is indispensable for evaluating the test sites as well as identifying genotypes with consistently specific and wider adaptation to particular agro-climatic zones. The genotypes identified in the present study could be used for commercial cultivation across edaphically challenged areas for sustainable production.

Smartphone, the New Learning Aid amongst Medical Students.

INTRODUCTION: The use of smartphone is increasing day by day for personal as well as professional purpose. They are becoming a more suitable tool for advancing education in developing countries. Mobile access to information and many applications are successfully harnessed in health care. Smartphones are also becoming popular as an effective educational tool. AIM: The present study was conducted to evaluate the use of smartphones as an educational tool amongst the medical students. The study also aimed at identifying the common medical application used by the students. MATERIALS AND METHODS: It was an observational cross-sectional study carried out amongst medical students of private medical institute in India. A validated 16 point, structured, open-ended, questionnaire regarding ownership and use of smart phones was self-administered to 446 medical students. Data were analysed using SPSS and open ended questions were analysed by summative content analysis. RESULTS: Among the study population, 96% owned a smartphone -Android based 72.4%, i phone 13.0%, Windows based Nokia phones 7% and Blackberry 3.6%. Common medical applications used by the students were Anatomy and Medical Dictionary in First MBBS; Medical Dictionary, Medscape and Google/Wikipedia in Second MBBS; and Medscape, Google/Wikipedia and Prognosis/Diagnosis in Third MBBS. More than 90% students, reported to have technological skills to use smartphones, for medical education, communication and instant access during bedside teaching. Advertently, 37.2% students felt if smartphones are used for clinical purposes, they will need to spend less time with patients. Almost 79.4% felt that smartphones should be introduced in MBBS course. CONCLUSION: Smartphone use amongst medical students as learning aid for various medical applications is rapidly advancing. But it will be worthwhile to study whether use of smartphones has any impact on the grades of the students before introducing them in medical schools.