Growth Suppression of Oral Squamous Cell Carcinoma Cells by Lactobacillus Acidophilus.

OBJECTIVES: Oral squamous cell carcinoma (OSCC) is a highly aggressive form of oral cancer. Probiotic lactobacilli have demonstrated anticancer effects, whilst their interaction with Streptococcus mutans in this context remains unexplored. The objective of this study was to investigate the antiproliferative effect of Lactobacillus acidophilus on OSCC and to understand the effect of S mutans on OSCCs and whether it affects the antiproliferative potential of L acidophilus when co-exposed to OSCC. METHODS: The human head and neck squamous cell carcinoma cells of the oral cavity (HNO97 cell line) were exposed to cultures of L acidophilus and S mutans separately and in combination. Further, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to assess the viability of HNO97 cells. Bacterial adhesion to HNO97 cells was examined by confocal microscopy and apoptosis by Nexin staining. To understand the underlying mechanism of apoptosis, expression of the tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) gene and protein were determined by real-time polymerase chain reaction and quantitative enzyme-linked immunosorbent assay, respectively. RESULTS: A significant decrease (53%-56%) in the viability of HNO97 cells on exposure to L acidophilus, S mutans, and the 2 species together demonstrated the antiproliferative activity of L acidophilus and S mutans. Both bacteria showed adhesion to HNO97 cells. The expression of the TRAIL gene increased 5-fold in HNO97 cells on treatment with L acidophilus and S mutans, which further increased to ∼17-fold with both species present. Expression levels of the TRAIL protein were significantly (P < .05) increased in bacteria-treated cell lysates. Further, bacteria-treated HNO97 cells exhibited lower live and intact cell percentages with higher proportions of cells in early and late apoptotic stages. CONCLUSIONS: L acidophilus exhibits the antiproliferative activity against OSCC cells possibly partially via a TRAIL-induced mechanism of apoptosis, which is not affected by the presence of S mutans. These findings may encourage further investigation into the possible therapeutic application of probiotic L acidophilus in OSCC.

Secretome analysis and virulence assessment in Abiotrophia defectiva.

Background: Abiotrophia defectiva, although infrequently occurring, is a notable cause of culture-negative infective endocarditis with limited research on its virulence. Associated with oral infections such as dental caries, exploring its secretome may provide insights into virulence mechanisms. Our study aimed to analyze and characterize the secretome of A. defectiva strain CCUG 27639. Methods: Secretome of A. defectiva was prepared from broth cultures and subjected to mass spectrometry and proteomics for protein identification. Inflammatory potential of the secretome was assessed by ELISA. Results: Eighty-four proteins were identified, with diverse subcellular localizations predicted by PSORTb. Notably, 20 were cytoplasmic, 12 cytoplasmic membrane, 5 extracellular, and 9 cell wall-anchored proteins. Bioinformatics tools revealed 54 proteins secreted via the 'Sec' pathway and 8 via a non-classical pathway. Moonlighting functions were found in 23 proteins, with over 20 exhibiting potential virulence properties, including peroxiredoxin and oligopeptide ABC transporter substrate-binding protein. Gene Ontology and KEGG analyses categorized protein sequences in various pathways. STRING analysis revealed functional protein association networks. Cytokine profiling demonstrated significant proinflammatory cytokine release (IL-8, IL-1ß, and CCL5) from human PBMCs. Conclusions: Our study provides a comprehensive understanding of A. defectiva's secretome, laying the foundation for insights into its pathogenicity.

Sugar Substitute Stevia Inhibits Biofilm Formation, Exopolysaccharide Production, and Downregulates the Expression of Streptococcal Genes Involved in Exopolysaccharide Synthesis.

BACKGROUND: Acid production by sucrose fermentation disturbs the balance in dental plaque by lowering the oral pH. As a consequence of the profound effect of sucrose on caries initiation and progression, many studies have been directed towards finding non-cariogenic artificial sweeteners that can be used as a substitute to sucrose. Existing literature shows that dietary sucrose upregulates the expression of biofilm associated genes involved in exopolysaccharide (EPS) production. OBJECTIVE: In this study, we aimed to investigate the effect of the sugar substitute stevia on biofilm formation, EPS secretion, and streptococcal genes encoding glucan-binding proteins (Gbps) and glucosyltransferases (Gtfs), which are essential for the synthesis of EPS. MATERIALS AND METHODS: Streptococcus mutans and Streptococcus gordonii were grown as biofilm cultures with or without stevia and sucrose. Biomass was quantified for biofilm and EPS production by crystal violet staining and the phenol-sulfuric acid method, respectively. Expression of gtfB and gbpB genes was studied by RT-PCR. RESULTS: The quantities of biofilm were significantly lower when grown in the presence of stevia compared to sucrose in both species (p < 0.05). The proportion of EPS in the biofilm pellet decreased with increasing concentrations of stevia in both species but remained nearly unchanged with sucrose with respect to the control. In both streptococcal species, exposure of stevia decreased the expression of gtfB and gbpB genes compared to sucrose (p < 0.05). In comparison to the untreated control, the expression was decreased in the presence of stevia in both species, while it increased 2.5- to 4-fold in S. mutans and 1.5- to 2.5-fold in S. gordonii in the presence of sucrose. CONCLUSION: The ability of stevia to inhibit biofilm formation, reduce EPS production, and downregulate the expression of gtfB and gbpB genes in S. mutans and S. gordonii may have potential therapeutic applications in controlling dental plaques and caries.

Screening of InDel variants in PRDM6, myostatin and IGF2BP1 genes and association analysis with body measurement traits in Malabari and Attappadi black goats.

An insertion/deletion (InDel) polymorphism study of PR/SET domain family 6 (PRDM6), myostatin (MSTN) and insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) genes was conducted in Malabari and Attappady black goats. An association study of identified InDels and body measurement traits was also performed. Body measurements included body length, chest diameter, chest depth, canon circumference, hip width, and hip height at the hip cross. The body trunk index, the body length index, the canon circumference index, and the chest width index were calculated. The Hardy-Weinberg equilibrium (HWE) was tested using a Chi-square test. The observed heterozygosity (Ho), expected heterozygosity (He), and polymorphism information content (PIC) were calculated. A significant difference in body measurements was found across breeds, ages, and breed x age interactions. The PRDM6 InDel was also associated with body measurement traits, such as body height, canon circumference and canon circumference index. In both Malabari and Attappadi black MSTN and PRDM6 InDels were in a state of HWE, while IGF2BP1 InDels were not. Indel markers found in the present study may be used for marker-assisted selection of growth traits among goats.

Genetic diversity analysis of cattle genetic groups of Kerala state using microsatellite data.

Cattle belonging to seven different genetic groups in Kerala state, India were chosen for the study to find out the genetic diversity between the groups, which would aid in their sustainable improvement and conservation of native cattle. They included the native groups namely, Vechur, Kasaragod, Vadakara dwarf and Vilwadri, along with three different grades of crossbred cattle, based on milk production. Genomic DNA was isolated from 20 to 30 unrelated animals of each group and a panel of 25 microsatellite markers as suggested by FAO-ISAG, were amplified by multiplex PCR. The PCR amplicons were genotyped and the allelic data analyzed using suitable Bioinformatics softwares. The present study showed that the observed number of alleles was much more than the expected, in all populations. The mean PIC value obtained for the present study was 0.8912 and increased number of private alleles were observed, especially in Vilwadri and Kasaragod groups. Negative value of FIS (-0.055) indicated that the level of inbreeding was less. The FST value was 0.1442 indicating that the populations showed good genetic differentiation. The results of Structure analysis revealed admixture only in Vadakara population. The results obtained from the present study showed that Vilwadri and Kasaragod cattle showed distinct differences from other groups.

Comparison of performance indicators of MGIT primary culture to assess the impact of implementation of ISO 15189:2012 standards.

Performance indicators are key component and plays a major role for monitoring and continuous quality improvement of the test results. The NABL certificate of accreditation is issued in accordance with the standard ISO 15189:2012 requirements. As part of the accreditation process, the laboratory has acquired knowledge and implemented the quality system procedures. Present study analyzed the impact of the accreditation process on the "performance indicators" of MGIT primary culture and found that performance indicators have been improved significantly after implementation of NABL for almost all indicators which clearly indicate the importance of accreditation and implementation of quality procedures for reliability of valid test results.

Proteomic analysis of the periodontal pathogen Prevotella intermedia secretomes in biofilm and planktonic lifestyles.

Prevotella intermedia is an important species associated with periodontitis. Despite the remarkable clinical significance, little is known about the molecular basis for its virulence. The aim of this study was to characterize the secretome of P. intermedia in biofilm and planktonic life mode. The biofilm secretome showed 109 proteins while the planktonic secretome showed 136 proteins. The biofilm and the planktonic secretomes contained 17 and 33 signal-peptide bearing proteins, 13 and 18 lipoproteins, respectively. Superoxide reductase, sensor histidine kinase, C40 family peptidase, elongation factor Tu, threonine synthase etc. were unique to biofilm. Of the ~ 30 proteins with predicted virulence potential from biofilm and planktonic secretomes, only 6 were common between the two groups, implying large differences between biofilm and planktonic modes of P. intermedia. From Gene Ontology biofilm secretome displayed a markedly higher percent proteins compared to planktonic secretome in terms of cellular amino acid metabolic process, nitrogen compound metabolic process etc. Inflammatory cytokine profile analysis revealed that only the biofilm secretome, not the planktonic one, induced important cytokines such as MIP-1α/MIP-1ß, IL-1ß, and IL-8. In conclusion, the revealed differences in the protein profiles of P. intermedia biofilm and planktonic secretomes may trigger further questions about molecular mechanisms how this species exerts its virulence potential in the oral cavity.

The Effect of Benzyl Isothiocyanate on the Expression of Genes Encoding NADH Oxidase and Fibronectin-Binding Protein in Oral Streptococcal Biofilms.

Recent studies have shown that antimicrobial treatment results in up- or down regulation of several virulence-associated genes in bacterial biofilms. The genes encoding NADH oxidase (nox) and fibronectin-binding protein (fbp) are known to play important roles in biofilm growth of some oral bacterial species. The objective was to study the effect of benzyl isothiocyanate (BITC), an antimicrobial agent from Miswak plant, on the expression of nox and fbp genes in some oral streptococci. The biofilms were treated with BITC and mRNA expression of nox and fbp genes was measured by comparative ΔΔCt method. The highest amount of biofilm mass was produced by A. defectiva, followed by S. gordonii, S. mutans, G. elegans and G. adiacens. Upon treatment with BITC, S. gordonii biofilms showed highest folds change in mRNA expression for both fbp and nox genes followed by S. mutans, A. defectiva, and G. adiacens. G. elegans mRNA levels for nox were extremely low. In conclusion, BITC treatment of the biofilms caused an upregulation of biofilm-associated genes fbp and nox genes in most of the tested species suggesting the significance of these genes in biofilm lifestyle of these oral bacteria and needs further investigation to understand if it contributes to antimicrobial resistance.

Optimization of multiple parameters for adsorption of arsenic (III) from aqueous solution using Psidium guajava leaf powder.

The conventional method of water treatment using activated carbon from several sources has been focused on extensively in the last two decades. However, rare attention has been noticed on natural adsorbents such as plant leaves. Therefore, the Psidium guajava (guava) leaf has been investigated to understand its adsorption efficacy for Arsenic (III) [As(III)] in this study. The effect of process variables, e.g., pH, concentration of metal ion, adsorbent's particle size, and dosages, are evaluated. Experiments are carried out in batch mode, and the individual and combined parameter's impact on adsorption have been discussed. Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) is used to characterize the adsorbent's surface. Freundlich and Langmuir's isotherms are used for adsorption equilibrium study. The adsorption parameters are optimized by establishing a regression correlation using central composite design (CCD) of response surface methodology (RSM). The analysis of variance (ANOVA) suggests a high regression coefficient (R2 = 0.9249) for the removal of As(III). Particle size of 0.39 mm; adsorbent's height of 10 cm; metal ion concentration of 30 ppm, and pH 6 are optimized to remove 90.88% As(III) from aqueous solution. HCl is evaluated as a potential solvent for desorption of arsenic from the desorption study.

Molecular characterisation of coding regions of HIF-1a gene in Vechur cattle by cDNA sequencing.

Hypoxia-inducible factor (HIF)-1α is a transcription factor stabilized by hypoxia by inducing or suppressing the homeostatic regulatory gene expression, enabling tissues and cells to survive despite fluctuations in environmental circumstances. As the name implies, hypoxia-inducible factor-1 is secreted not only as a cellular response to hypoxia but also in heat stress and oxidative stress. The goal of this work was to determine the molecular characterisation of the HIF-1α gene coding region as well as the differences in HIF-1αprotein primary structure between Vechur cattle and other cattle breeds in the online databases. Total RNA was isolated from blood samples of 6 Vechur cattle using the trizol reagent method, and full-length c sequences of the HIF-1α gene were sequenced. The base pair length of composite HIF-1αcDNA of Vechur cattle and encoding ORFis 3956 bp and 2469 bp respectively. The 5'UTR was recognized to be 279 bp in length. The start codon was identified at nucleotide 280-282, the stop codon UGA at 2746-2748 bp and a 1208 bp 3'UTR which included a poly-A tail of 27 adenine residues. In a comparative analysis of the cDNA, point transitions causing guanine to adenine (G>A) changes at 1211th and 2699th positions were noticed as a heterozygous condition in the whole 3956 bp sequence. These two SNVs in the coding regions were responsible for two amino acid changes in the deduced 823 amino acid sequence. Since the predicted amino acid arginine had been replaced with lysine at 311th and 807th positions, it showed 99.76 percent sequence identity with Bos taurus. The phylogenetic tree revealed that the HIF-1α protein of Vechur cattle had a lesser evolutionary distance from the same gene of related species emphasising the highly conserved nature of this particular protein. This structural variation observed in the present study should be evaluated on a larger population to assess its functional relevance for thermo-tolerance.

In vitro Characterization of Biofilm Formation in Prevotella Species.

Background: Periodontitis, a chronic inflammatory oral infection is the outcome of disturbances in the homeostasis of the oral biofilm microbiota. A number of studies have found the occurrence of Prevotella species in elevated levels in periodontitis compared to healthy subjects. Even though different aspects of Prevotella as part of oral biofilm have been studied, in vitro biofilms formed by these species have not been characterized systematically. The objective of this study was to characterize biofilms formed by several Prevotella species and further to assess biofilm inhibition and detachment of preformed biofilms. Methods: Biofilms were grown in 24-well plates containing brucella broth in anaerobic conditions for 3 days, and were quantified using crystal violet staining. Images of SYTO 9 Green fluorescent stained biofilms were captured using confocal microscopy. Biofilm inhibition and detachment by proteinase and DNase I was tested. The biochemical characterization included quantification of proteins and DNA in the biofilms and biofilm-supernatants. Results: Prevotella loescheii, Prevotella oralis and Prevotella nigrescens showed highest biofilm formation. P. nigrescens formed significantly higher amounts of biofilms than P. loescheii (P = 0.005) and P. oralis (P = 0.0013). Inhibition of biofilm formation was significant only in the case of P. oralis when treated with proteinase (P = 0.037), whereas with DNase I treatment, the inhibition was not significant (P = 0.531). Overall, proteinase was more effective in biofilm detachment than DNase I. Protein and DNA content were higher in biofilm than the supernatant with the highest amounts found in P. nigrescens biofilm and supernatants. P. oralis biofilms appeared to secrete large amounts of proteins extracellularly into the biofilm-supernatants. Conclusion: Significant differences among Prevotella species to form biofilms may imply their variable abilities to get integrated into oral biofilm communities. Of the species that were able to grow as biofilms, DNase I and proteinase inhibited the biofilm growth or were able to cause biofilm detachment.

Ovarian expression, polymorphism identification and association of SIRT3 gene with reproduction traits in goats.

The NAD+-dependent protein deacetylase, Sirtuin3 (SIRT3), plays a role in fertility by preventing the activation of reactive oxygen species (ROS). A novel study was conducted on caprine SIRT3, to study its ovarian expression, explore the sequence variability in exon 7 and analyze its association with prolificacy in two native goat breeds of Kerala, Malabari and Attappady Black. The mRNA isolated from ovaries of six Malabari and Attappady Black goats were subjected to quantitative PCR (qPCR) using GAPDH and ß-actin as reference genes. Genomic DNA was isolated from 185 goats (99 Malabari and 86 Attappady Black) and subjected to PCR-SSCP to identify polymorphism in exon 7 of SIRT3 and association with litter size was analyzed. The ovarian expression of caprine SIRT3 was significantly higher (p ≤ 0.01) in Malabari than low prolific Attappady Black. PCR-SSCP analysis revealed, exon 7 of SIRT3 was polymorphic with three genotypes namely, AA, AB and BB with a novel SNP, g.154C > T in the 3'UTR. A significant association (p ≤ 0.05) was noticed between the genotypes of SIRT3 and litter size. The results obtained from this study highlight the role of SIRT3 in reproduction and hence SIRT3 may be considered as a potential candidate gene for genetic improvement in goats.

Proteomics of extracellular vesicles produced by Granulicatella adiacens, which causes infective endocarditis.

When oral bacteria accidentally enter the bloodstream due to transient tissue damage during dental procedures, they have the potential to attach to the endocardium or an equivalent surface of an indwelling prosthesis and cause infection. Many bacterial species produce extracellular vesicles (EVs) as part of normal physiology, but also use it as a virulence strategy. In this study, it was hypothesized that Granulicatella adiacens produce EVs that possibly help it in virulence. Therefore, the objectives were to isolate and characterize EVs produced by G. adiacens and to investigate its immune-stimulatory effects. The reference strain G. adiacens CCUG 27809 was cultured on chocolate blood agar for 2 days. From subsequent broth culture, the EVs were isolated using differential centrifugation and filtration protocol and then observed using scanning electron microscopy. Proteins in the vesicle preparation were identified by nano LC-ESI-MS/MS. The EVs proteome was analyzed and characterized using different bioinformatics tools. The immune-stimulatory effect of the EVs was studied via ELISA quantification of IL-8, IL-1ß and CCL5, major proinflammatory cytokines, produced from stimulated human PBMCs. It was revealed that G. adiacens produced EVs, ranging in diameter from 30 to 250 nm. Overall, G. adiacens EVs contained 112 proteins. The proteome consists of several ribosomal proteins, DNA associated proteins, binding proteins, and metabolic enzymes. It was also shown that these EVs carry putative virulence factors including moonlighting proteins. These EVs were able to induce the production of IL-8, IL-1ß and CCL5 from human PBMCs. Further functional characterization of the G. adiacens EVs may provide new insights into virulence mechanisms of this important but less studied oral bacterial species.

Biofilm growth and IL-8 & TNF-α-inducing properties of Candida albicans in the presence of oral gram-positive and gram-negative bacteria.

BACKGROUND: Interaction of C. albicans with oral bacteria is crucial for its persistence, but also plays a potential role in the infection process. In the oral cavity, it grows as part of dental plaque biofilms. Even though growth and interaction of C. albicans with certain bacterial species has been studied, little is known about its biofilm growth in vitro in the simultaneous presence of Gram-negative and Gram-positive bacteria. The aim was to evaluate the growth of C. albicans in polymicrobial biofilms comprising oral Gram-negative and Gram-positive bacteria. Further, we also aimed to assess the potential of C. albicans in the Candida-bacteria polymicrobial biofilm to elicit cytokine gene expression and cytokine production from human blood cells. RESULTS: C. albicans cell counts increased significantly up to 48 h in polymicrobial biofilms (p < 0.05), while the bacterial counts in the same biofilms increased only marginally as revealed by qPCR absolute quantification. However, the presence of bacteria in the biofilm did not seem to affect the growth of C. albicans. Expression of IL-8 gene was significantly (p < 0.05) higher upon stimulation from biofilm-supernatants than from biofilms in polymicrobial setting. On the contrary, TNF-α expression was significantly higher in biofilms than in supernatants but was very low (1-4 folds) in the monospecies biofilm of C. albicans. ELISA cytokine quantification data was in agreement with mRNA expression results. CONCLUSION: Persistence and enhanced growth of C. albicans in polymicrobial biofilms may imply that previously reported antagonistic effect of A. actinomycetemcomitans was negated. Increased cytokine gene expression and cytokine production induced by Candida-bacteria polymicrobial biofilms and biofilm supernatants suggest that together they possibly exert an enhanced stimulatory effect on IL-8 and TNF-α production from the host.

Real-time polymerase chain reaction quantification of the salivary levels of cariogenic bacteria in patients with orthodontic fixed appliances.

AIM: The aim was to investigate the salivary detection frequencies and quantities of caries-associated bacteria from patients with orthodontic brackets. METHODS: Patients wearing orthodontic brackets (n = 40, mean age = 26 years) and healthy controls without brackets (n = 40, mean age = 17 years) were enrolled in the study. Saliva samples from each patient was collected. After DNA purification, target species comprising streptococci and a Lactobacillus species were detected and quantified from the samples using polymerase chain reaction (PCR) and real-time quantitative PCR. RESULTS: Detection frequencies did not differ between the orthodontic patients and the control subjects for any target species except for Streptococcus sobrinus, which showed significantly lower detection rates in orthodontic patients (p < .05). Lactobacillus casei and Streptococcus gordonii were found at the highest detection frequencies with both species being detected in 38 (95%) of the saliva samples of orthodontic patients. Similarly, L. casei and Streptococcus salivarius were the species with highest detection frequencies (35, 87.5%) in the control subjects. Real-time PCR revealed that Streptococcus mutans and S. salivarius quantities were significantly higher in orthodontic patients than in the control subjects (p < .05). CONCLUSIONS: Application of orthodontic brackets for 12 months leads to increased salivary levels of cariogenic bacteria and may serve as a potential risk factor for caries initiation.

Metagenomic analysis exploring taxonomic diversity of rumen microbial communities in Vechur and crossbred cattle of Kerala state, India.

Rumen, one of the most productive diverse microbial habitats plays a vital role in the breakdown of feed to produce energy for maintenance and milk production in cattle. Culture-based procedures could identify only 10% of microbial species present in the rumen. Kerala, one of the southern states of India, owns only one native cattle breed, the Vechur cattle, which is noted for its short stature, disease resistance and adaptability to hot humid climate. Lower population density and decreased milk production potential of Vechur cattle led to the development of crossbred cattle of Kerala, with higher milk yield. A study was conducted to assess the rumen microbial profile of low productive Vechur cattle and high productive crossbred cattle for a better understanding of the relationship between the host and microbial community. DNA isolated from rumen liquor of five cattle each from both genetic groups maintained on standard ration (forage, concentrate ratio of 50:50) was subjected to whole metagenome sequencing. Bioinformatics and statistical analysis revealed that bacteria followed by archaea and eukaryota dominated in the Vechur cattle as well as the crossbred cattle rumen. Bacterial community was dominated by Bacteroidetes and Firmicutes phyla in both genetic groups with a higher Firmicutes to Bacteroidetes ratio of 0.45 in Vechur cattle. Among archaea, Euryarchaeota was more abundant, which constitute methanogens, contributing 98% of total archaeal reads. Prevalent protozoal genus found in the Vechur cattle rumen was Entodinium and in crossbred cattle rumen was Entamoeba. In Vechur and crossbred cattle rumen, 1086 and 1262 microbial species were observed exclusively and 4731 species were shared between habitats. There was a significant difference in total microbial species abundance between the two genetic groups and Vechur cattle displayed significantly higher microbial diversity compared to crossbred. As per literature, this is presumably the first report of rumen metagenome profile of Vechur cattle, a unique short breed of India.

Endoscopic assisted removal - An aesthetic approach to impacted third molar tooth in the orbital floor.

A third molar displaced in the orbital floor is a very rare sighting. The usual surgical approach to such a case is the Caldwell Luc procedure that has its own complications. Here in this article, we present a 17-year-old male patient with third molar displaced in the orbital floor with egg shell thin anterior wall. The aim of the article is to report the unusuality of the case and to emphasize the endoscopic assisted approach to the orbital floor through a relatively small sub-labial incision. Endoscope aids in preserving the integrity of vital structures, facial aesthetics and complete eradication of the disease with faster recovery. Though, endoscopic approach requires requisite expertise, it is a learnable skill and can be mastered with ease.

An in silico evaluation of non-synonymous single nucleotide polymorphisms of mastitis resistance genes in cattle.

Recognition of deleterious non-synonymous single nucleotide polymorphism (SNPs) aids in the assessment of genetic basis of diseases and prediction of clinical phenotypes. In this study, data obtained from whole exome sequencing of Vechur cow using Illumina HiSeq 2500 platform is compared with that of crossbred cattle of Kerala. Sequence analysis of selected 18 mastitis resistant genes, evaluated the consequence of non-synonymous SNPs in these genes from both Vechur and crossbred cattle of Kerala, using sequence and structure-based computational tools such as SIFT, PROVEAN and I-MUTANT 2.0. Compared to Vechur cattle, incidence of missense deleterious mutation to effect protein functioning were relatively higher in crossbred cattle. These results on the type of genetic variants and its impact on normal functioning of a protein will assist to predict and enhance the disease resistance in cattle breeds.

Proteomic Analysis and Virulence Assessment of Granulicatella adiacens Secretome.

Despite reports on the occurrence of Granulicatella adiacens in infective endocarditis, few mechanistic studies on its virulence characteristics or pathogenicity are available. Proteins secreted by this species may act as determinants of host-microbe interaction and play a role in virulence. Our aim in this study was to investigate and functionally characterize the secretome of G. adiacens. Proteins in the secretome preparation were digested by trypsin and applied to nanoLC-ESI-MS/MS. By using a combined mass spectrometry and bioinformatics approach, we identified 101 proteins. Bioinformatics tools predicting subcellular localization revealed that 18 of the secreted proteins possessed signal sequence. More than 20% of the secretome proteins were putative virulence proteins including serine protease, superoxide dismutase, aminopeptidase, molecular chaperone DnaK, and thioredoxin. Ribosomal proteins, molecular chaperones, and glycolytic enzymes, together known as "moonlighting proteins," comprised fifth of the secretome proteins. By Gene Ontology analysis, more than 60 proteins of the secretome were grouped in biological processes or molecular functions. KEGG pathway analysis disclosed that the secretome consisted of enzymes involved in biosynthesis of antibiotics. Cytokine profiling revealed that secreted proteins stimulated key cytokines, such as IL-1ß, MCP-1, TNF-α, and RANTES from human PBMCs. In summary, the results from the current investigation of the G. adiacens secretome provide a basis for understanding possible pathogenic mechanisms of G. adiacens.

Evaluation of key survey components of bio-behavioral surveillance among HIV high-risk subpopulation in Western India.

BACKGROUND: High-quality data are of prime importance in any health survey because survey data are considered as a gold standard for nationally representative data. The quality of data collection largely depends on the design of the questionnaire, training, and skills of the interviewer. OBJECTIVES: In the present study, we tried to evaluate three key components, such as questionnaire design, human resource and training of the field staff for Integrated Biological and Behavioural Surveillance carried out among the HIV high-risk subpopulation. METHODS: A mixed-methods approach was used. Qualitative and quantitative data collection was carried out in the year 2015 with cross-sectional survey design in western states of India. The in-depth interviews of 10 stakeholders, structured interviews of the survey respondents (n = 560), and field investigators (n = 71) were conducted. Data triangulation was used to find out the concurrence of the qualitative and quantitative data. RESULTS: Comprehensive and standardized survey questionnaire, structured training agenda, and strategic preparation for recruiting human resources were the overall strengths of the survey. However, during the implementation of the survey, there were some difficulties reported in data collection process. Overall, the respondents and investigators felt that the questionnaire was long and exhaustive. Difficulties were faced while collecting data on sexual history. The field staffs were not adequately experienced to work with sensitive population. CONCLUSIONS: In order to have accurate, reliable data, especially on sexual behavior; emphasis should be given on simple questionnaire with the use of community-friendly language, skilled and experienced interviewers for data collection, and extensive field training.