Publications by authors named "Kishore G Bhat"

27 Publications

  • Page 1 of 1

Green tea catechins showed antibacterial activity on streptococcus mutans -An study.

Indian J Dent Res 2021 Apr-Jun;32(2):226-229

Department of Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India.

Background: The main bacterial aetiological agents in caries formation are the α-haemolytic Streptococcal species Streptococcus mutans, which has been found to be the initiator of most dental caries. The leaves of Camellia sinensis known as green tea, has properties, such as antibacterial and anti-cariogenic. Epigallocatechin-3-gallate (EGCG) one of the most abundant catechins found in green tea is known to contribute to these effects.

Aim: To evaluate the antibacterial effect of green tea catechins namely EGCG on S. mutans with two different methods at different concentrations.

Objectives: 1) To assess the antimicrobial efficacy of EGCG by disc diffusion test at concentrations of 100, 75, and 50 μg/mL. 2) To assess the antimicrobial efficacy of EGCG by Minimum inhibitory concentration (MIC) test at concentrations ranging from 0.2 to 100 μg/mL.

Methodology: Commercially available purest form of green tea polyphenol EGCG was used in the study. Disc diffusion test on agar medium and MIC test was used to determine the susceptibility of the S. mutans to green tea catechins EGCG.

Results: The results of the agar well diffusion method showed that the EGCG extract has shown zones of inhibition against S. mutans at concentrations of 100 μg/mL (28.67 mm), 75 μg/mL (15.33 mm), 50 μg/mL (10.33 mm) while that of MIC test of EGCG extract of concentrations ranging from 0.2 to 100 μg/mL against S. mutans shows that the mean MIC value was 1.07.

Conclusion: Catechins in the tea are potentially anti-cariogenic agents which can reduce bacterial presence in the oral cavity and have the potential to be further used for the preparation of dentifrice and mouthwash.
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http://dx.doi.org/10.4103/ijdr.ijdr_512_21DOI Listing
November 2021

Solitary Median Maxillary Central Incisor: A Case Report with 3-Year Follow-Up and Literature Review.

Contemp Clin Dent 2021 Jul-Sep;12(3):324-327. Epub 2021 Sep 21.

Department of Microbiology, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belgaum, Karnataka, India.

A solitary median maxillary central incisor (SMMCI) is a rare anomaly that can occur alone or be associated with other systemic abnormalities. Early diagnosis of SMMCI is crucial as it might indicate the presence of an associated congenital or developmental abnormality. The prevalence of live-born children with SMMCI is determined to be 1:50,000 and is more common among females. The purpose of this paper was to report an unusual case of a 9-year-old girl with SMMCI who had no growth deficiency or any other systemic involvement. Since pediatricians and dentists are the first professionals to evaluate an SMMCI's patient in most cases, it is important that they be aware of the possibility of other related systemic problems that require systemic care. Appropriate treatment, diagnosis, and referral should also include neuropediatric evaluation, genetic testing, and craniofacial profile analysis along with multidisciplinary approach.
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http://dx.doi.org/10.4103/ccd.ccd_713_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8525815PMC
September 2021

Cancer Stem Cell Traits in Tumor Spheres Derived from Primary Laryngeal Carcinoma Cell Lines.

Contemp Clin Dent 2021 Jul-Sep;12(3):247-254. Epub 2021 Sep 21.

Department of Oral and Maxillofacial Surgery, Maratha Mandal's N. G. Halgekar Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India.

Objective: Cancer stem cells (CSCs) belong to a subpopulation of undifferentiated cells present within tumors that have the potential to regenerate, differentiate, maintenance of pluripotency, drug resistance, and tumorigenicity when transplanted into an innate host. These can influence the growth and behavior of these tumors and are used to investigate the initiation, progression, and treatment strategies of laryngeal cancer. Research on CSC science and targeted therapies were hinge on their isolation and/or enrichment procedures. The object of the study is to isolate cancer stem cells from primary laryngeal carcinoma (CSCPLC) by tumor spheres enrichment. We checked the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance.

Materials And Methods: We performed tumor sphere formation assay (primary, secondary, and tertiary) chemotherapy resistance by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were performed to evaluate the CSC cells. Immunofluorescence for stem cell markers (CD133+, CD44+) and gene expression of stem cell markers for CD133+, CD44+, OCT4, SOX2, and NANOG was done using the real-time polymerase chain reaction technique.

Results: We were able to isolated CSC subpopulations from PLC cell lines by the tumor sphere method. These cells exhibited good primary, secondary, and tertiary tumor sphere formation efficiency and also disclosed a resistant index of more than 2. Immunofluorescence for stem cell markers (CD133+ and CD44+) confirms the presence of CSC. There was significantly higher mRNA expression of stem cell markers in CSC enriched subpopulations compared to the parental cell lines.

Conclusion: We conclude that tumor spheres enrichment is an efficient, economical, and reliable approach for the isolation and characterization of CSC from PLC cell lines. These cells demonstrated the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance.
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http://dx.doi.org/10.4103/ccd.ccd_252_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8525812PMC
September 2021

Coculture method for cultivation of uncultured oral bacteria.

J Oral Maxillofac Pathol 2021 May-Aug;25(2):266-271. Epub 2021 Aug 31.

Central Research Laboratory, Maratha Mandal's Nathajirao G Halgekar Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India.

Purpose: The purpose of the study is to culture uncultured oral bacteria with helper strains using the coculture method from the subgingival plaque samples of chronic periodontitis patients.

Materials And Methods: The samples were processed and inoculated on a blood agar medium enriched with hemin and Vitamin K. A helper strain (ATCC 6919) was cross-streaked across the inoculums to facilitate coculture. The plates were then incubated for 7 days with subsequent subculturing and further incubation.

Results: Satellite colonies around helper strain showed one colony type of Porphyromonas gingivalis, one was of nonpigmented , three were of and five isolates remained unidentified.

Conclusions: Coculture could be used effectively as one of the methods in the isolation and cultivation of oral bacteria. Incubation using the anaerobic jar technique was found to be economical and efficient for the growth of anaerobic oral bacteria.
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http://dx.doi.org/10.4103/0973-029X.325125DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8491346PMC
August 2021

Antimicrobial susceptibility pattern of oral gram negative anaerobes from Indian subjects.

Anaerobe 2021 Aug 17;70:102367. Epub 2021 Apr 17.

Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Bauxite Road, Belgaum, Karnataka, India.

Objectives: There is paucity of information on the antimicrobial susceptibility pattern of oral anaerobic bacteria. In this study, an attempt has been made to evaluate the antimicrobial susceptibility/resistance trend of oral Gram negative bacteria from Indian subjects.

Methods: Minimum inhibitory concentrations (MIC) of 304 isolates against twelve different antibiotics were determined using gradient diffusion MIC strips. The organisms were isolated and identified based on phenotypic characteristics and included Porphyromonas gingivalis, Prevotella species, Tannerella forsythia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcoitans, Eickenella corrodens and Capnocytophaga species. For each antimicrobial agent, MIC and MIC were calculated and expressed.

Results: Resistance to azithromycin, clindamycin, and amoxicillin was observed in most of the anaerobic bacterial species studied. High degree of susceptibility was observed to amoxillin-clavulanic acid, doxycycline and moxifloxacin. A single strain of P. melaninogenica was resistant to moxifloxacin. The susceptibility pattern varied with cephalosporins among species. Ceftriaxone showed highest and cefazolin least efficacy among cephalosporins. All anaerobic bacteria tested were susceptible to metronidazole. Strains of T. forsythia were more resistant to several antibiotics than other anaerobic bacteria. All three species of capnophilic bacteria displayed high degree of resistance to metronidazole and significant resistance to amoxicillin, azithromycin, clindamycin, cefazolin and cefuroxime.

Conclusions: Amoxicillin-clavulanic acid, doxycycline, moxifloxacin and metronidazole appeared to be the most effective drugs against gram negative anaerobic bacteria. However, the MIC and MIC values against metronidazole were on the higher side of the normal indicating a potential for developing resistance.
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http://dx.doi.org/10.1016/j.anaerobe.2021.102367DOI Listing
August 2021

Adjunctive systemic antimicrobials for the non-surgical treatment of periodontitis.

Cochrane Database Syst Rev 2020 11 16;11:CD012568. Epub 2020 Nov 16.

Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.

Background: Systemic antimicrobials can be used as an adjunct to mechanical debridement (scaling and root planing (SRP)) as a non-surgical treatment approach to manage periodontitis. A range of antibiotics with different dosage and combinations are documented in the literature. The review follows the previous classification of periodontitis as all included studies used this classification.

Objectives: To assess the effects of systemic antimicrobials as an adjunct to SRP for the non-surgical treatment of patients with periodontitis.

Search Methods: Cochrane Oral Health's Information Specialist searched the following databases to 9 March 2020: Cochrane Oral Health's Trials Register, CENTRAL, MEDLINE, and Embase. The US National Institutes of Health Ongoing Trials Register ClinicalTrials.gov and the World Health Organization International Clinical Trials Registry Platform were searched for ongoing trials.

Selection Criteria: We included randomized controlled trials (RCTs) which involved individuals with clinically diagnosed untreated periodontitis. Trials compared SRP with systemic antibiotics versus SRP alone/placebo, or with other systemic antibiotics.

Data Collection And Analysis: We selected trials, extracted data, and assessed risk of bias in duplicate. We estimated mean differences (MDs) for continuous data, with 95% confidence intervals (CIs). We assessed the certainty of the evidence using GRADE.

Main Results: We included 45 trials conducted worldwide involving 2664 adult participants. 14 studies were at low, 8 at high, and the remaining 23 at unclear overall risk of bias. Seven trials did not contribute data to the analysis. We assessed the certainty of the evidence for the 10 comparisons which reported long-term follow-up (≥ 1 year). None of the studies reported data on antimicrobial resistance and patient-reported quality of life changes. Amoxicillin + metronidazole + SRP versus SRP in chronic/aggressive periodontitis: the evidence for percentage of closed pockets (MD -16.20%, 95% CI -25.87 to -6.53; 1 study, 44 participants); clinical attachment level (CAL) (MD -0.47 mm, 95% CI -0.90 to -0.05; 2 studies, 389 participants); probing pocket depth (PD) (MD -0.30 mm, 95% CI -0.42 to -0.18; 2 studies, 389 participants); and percentage of bleeding on probing (BOP) (MD -8.06%, 95% CI -14.26 to -1.85; 2 studies, 389 participants) was of very low certainty. Only the results for closed pockets and BOP showed a minimally important clinical difference (MICD) favouring amoxicillin + metronidazole + SRP. Metronidazole + SRP versus SRP in chronic/aggressive periodontitis: the evidence for percentage of closed pockets (MD -12.20%, 95% CI -29.23 to 4.83; 1 study, 22 participants); CAL (MD -1.12 mm, 95% CI -2.24 to 0; 3 studies, 71 participants); PD (MD -1.11 mm, 95% CI -2.84 to 0.61; 2 studies, 47 participants); and percentage of BOP (MD -6.90%, 95% CI -22.10 to 8.30; 1 study, 22 participants) was of very low certainty. Only the results for CAL and PD showed an MICD favouring the MTZ + SRP group. Azithromycin + SRP versus SRP for chronic/aggressive periodontitis: we found no evidence of a difference in percentage of closed pockets (MD 2.50%, 95% CI -10.19 to 15.19; 1 study, 40 participants); CAL (MD -0.59 mm, 95% CI -1.27 to 0.08; 2 studies, 110 participants); PD (MD -0.77 mm, 95% CI -2.33 to 0.79; 2 studies, 110 participants); and percentage of BOP (MD -1.28%, 95% CI -4.32 to 1.76; 2 studies, 110 participants) (very low-certainty evidence for all outcomes). Amoxicillin + clavulanate + SRP versus SRP for chronic periodontitis: the evidence from 1 study, 21 participants for CAL (MD 0.10 mm, 95% CI -0.51 to 0.71); PD (MD 0.10 mm, 95% CI -0.17 to 0.37); and BOP (MD 0%, 95% CI -0.09 to 0.09) was of very low certainty and did not show a difference between the groups. Doxycycline + SRP versus SRP in aggressive periodontitis: the evidence from 1 study, 22 participants for CAL (MD -0.80 mm, 95% CI -1.49 to -0.11); and PD (MD -1.00 mm, 95% CI -1.78 to -0.22) was of very low certainty, with the doxycycline + SRP group showing an MICD in PD only. Tetracycline + SRP versus SRP for aggressive periodontitis: we found very low-certainty evidence of a difference in long-term improvement in CAL for the tetracycline group (MD -2.30 mm, 95% CI -2.50 to -2.10; 1 study, 26 participants). Clindamycin + SRP versus SRP in aggressive periodontitis: we found very low-certainty evidence from 1 study, 21 participants of a difference in long-term improvement in CAL (MD -1.70 mm, 95% CI -2.40 to -1.00); and PD (MD -1.80 mm, 95% CI -2.47 to -1.13) favouring clindamycin + SRP. Doxycycline + SRP versus metronidazole + SRP for aggressive periodontitis: there was very low-certainty evidence from 1 study, 27 participants of a difference in long-term CAL (MD 1.10 mm, 95% CI 0.36 to 1.84); and PD (MD 1.00 mm, 95% CI 0.30 to 1.70) favouring metronidazole + SRP. Clindamycin + SRP versus metronidazole + SRP for aggressive periodontitis: the evidence from 1 study, 26 participants for CAL (MD 0.20 mm, 95% CI -0.55 to 0.95); and PD (MD 0.20 mm, 95% CI -0.38 to 0.78) was of very low certainty and did not show a difference between the groups. Clindamycin + SRP versus doxycycline + SRP for aggressive periodontitis: the evidence from 1 study, 23 participants for CAL (MD -0.90 mm, 95% CI -1.62 to -0.18); and PD (MD -0.80 mm, 95% CI -1.58 to -0.02) was of very low certainty and did not show a difference between the groups. Most trials testing amoxicillin, metronidazole, and azithromycin reported adverse events such as nausea, vomiting, diarrhoea, mild gastrointestinal disturbances, and metallic taste. No serious adverse events were reported.

Authors' Conclusions: There is very low-certainty evidence (for long-term follow-up) to inform clinicians and patients if adjunctive systemic antimicrobials are of any help for the non-surgical treatment of periodontitis. There is insufficient evidence to decide whether some antibiotics are better than others when used alongside SRP. None of the trials reported serious adverse events but patients should be made aware of the common adverse events related to these drugs. Well-planned RCTs need to be conducted clearly defining the minimally important clinical difference for the outcomes closed pockets, CAL, PD, and BOP.
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http://dx.doi.org/10.1002/14651858.CD012568.pub2DOI Listing
November 2020

Effect of curcumin on growth, biofilm formation and virulence factor gene expression of Porphyromonas gingivalis.

Odontology 2021 Jan 11;109(1):18-28. Epub 2020 Apr 11.

Central Research Laboratory, Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences and Research Centre, Near KSRP Ground, Bauxite road, Belgaum, Karnataka, 590 010, India.

Porphyromonas gingivalis is a keystone pathogen and major colonizer in host tissue which plays a pivotal role in periodontitis among the other polymicrobial infections. Increasing facts demonstrate that curcumin has antibacterial activity and anti-biofilm effect against the periodontopathogens through diverse mechanisms that have a positive impact on periodontal health. The present study was aimed to elucidate the effect of curcumin on biofilm formation and virulence factor gene expression of P. gingivalis. By using gene expression studies, we exploited the mechanism of anti-biofilm effects of curcumin on P. gingivalis. The minimum inhibitory concentration and minimum bactericidal concentration of curcumin for both ATCC and clinical strains of P. gingivalis were found to be 62.5 and 125 µg ml respectively. Curcumin prevented bacterial adhesion and biofilm formation in a dose-dependent manner. Further, curcumin attenuated the virulence of P. gingivalis by reducing the expression of genes coding for major virulence factors, including adhesions (fimA, hagA, and hagB) and proteinases (rgpA, rgpB, and kgp). The results indicated that curcumin has shown anti-biofilm as well as antibacterial activity against P. gingivalis. Further, curcumin because of its pleiotropic actions could be a simple and inexpensive therapeutic strategy in the treatment of periodontal disease.
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http://dx.doi.org/10.1007/s10266-020-00514-yDOI Listing
January 2021

Roles of the matricellular protein Tenascin-C in T-lymphocyte trafficking and etiopathogenesis of Oral Lichen Planus.

Arch Oral Biol 2020 Feb 20;110:104622. Epub 2019 Nov 20.

Oral Pathology and Microbiology, KLE's VK Institute of Dental Sciences and Hospital, Belgaum, Karnataka 590010, India; Oral Biology, School of Dental Medicine, University of Buffalo, New York, 14214, USA. Electronic address:

Objective: This study was aimed at examining the role of Tenascin-C in T cell trafficking in Oral Lichen Planus (OLP).

Design: For the in vivo immunohistochemical analyses, 115 OLP samples were collected from patients and immunostaining was performed. The intensity and distribution of TN-C expression were quantified and correlated with histological analyses of basement membrane integrity and presence of inflammatory infiltrate. For the in vitro study, TN-C and collagen were coated on culture plates and migration of T lymphocytes was assessed.

Results: TN-C immunoexpression was increased in terms of both distribution and intensity along the basement membrane zone. These changes were significantly associated with basement membrane duplication (distribution p < 0.002 and intensity p < 0.001) and bands of inflammation (distribution p < 0.002 and intensity p < 0.001) assessed by Chi-square test. T lymphocytes demonstrated significant migration towards TN-C as compared to collagen (n = 3, p < 0.05).

Conclusions: These findings indicate TN-C may have a key role in promoting T cell migration at the epithelial-mesenchymal junction in OLP. These observations suggest TN-C could be a good target for therapeutic intervention, either in itself or synergistically with anti-inflammatory directed strategies in this chronic disease management.
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http://dx.doi.org/10.1016/j.archoralbio.2019.104622DOI Listing
February 2020

Quantitative assessment of Scardovia wiggsiae from dental plaque samples of children suffering from severe early childhood caries and caries free children.

Anaerobe 2020 Apr 16;62:102110. Epub 2019 Oct 16.

Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India.

Scardovia wiggsiae has recently been identified as a potential pathogen associated with dental caries. The aim of the present study was to detect and quantify S. wiggsiae from dental plaque samples of children suffering from severe early childhood caries and children who were caries free by employing a real time DNA polymerase chain reaction (Real-time PCR) method. Dental plaque samples were collected from children suffering from severe early childhood caries (S-ECC) (n = 30) and caries free children (CF) (n = 30) reporting to the out-patient clinics of the department of paediatric and preventive dentistry. Plaque samples from each group were subjected to real-time PCR, post DNA extraction. Both the groups showed the presence of the organism S. wiggsiae, however there was a significant difference in its quantification between groups, with the median number being 1.49 × 10 cells per ml in caries free samples compared to 1.40 × 10 cells per ml in S-ECC samples. S. wiggsiae were isolated from nearly all samples of children, both caries free and those suffering from S-ECC. However, their numbers differ drastically in both groups with the scales tipping towards the S-ECC group, proving their association with the disease process in a significant manner. The present study shows significant association of S. wiggsiae in severe early childhood caries.
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http://dx.doi.org/10.1016/j.anaerobe.2019.102110DOI Listing
April 2020

evaluation of cytotoxicity of (amla) on cultured human primary dental pulp fibroblasts.

J Indian Soc Pedod Prev Dent 2019 Jul-Sep;37(3):251-257

Department of Microbiology and Molecular Biology, Maratha Mandal's NGH Institute of Dental Sciences and Research Center, Belagavi, Karnataka, India.

Context: The dental pulp tissue is capable of healing after surgical amputation of infected/inflamed tissue during vital pulp therapy, when in contact with a suitable medicament. Emblica officinalis (amla), a traditional medicine, is one such medicament which has never been evaluated for its healing potential in pulp therapy.

Aims: The aim of the study was to evaluate the cytotoxicity of E. officinalis (amla) against human primary dental pulp fibroblasts.

Settings And Design: This was in vitro study.

Subjects And Methods: Human dental pulp fibroblasts were obtained from dental pulp tissue of extracted over-retained primary incisors. The primary cells were cultured using the Dulbecco's Modified Eagle's Medium and used for the study after the fourth passage. The test medicament was E. officinalis with mineral trioxide aggregate (MTA) (100%) and untreated cells as positive and negative controls, respectively. Methyl-thiazol-diphenyl-tetrazolium (MTT) cytotoxicity assay was performed, and the cell survival was observed and analyzed at intervals of 24, 48, and 72 h.

Statistical Analysis Used: Cell survival within groups was compared with Wilcoxon matched-paired t-test and in between groups at each point interval was analyzed with the Kruskal-Wallis ANOVA test. The level of significance was set at 0.05.

Results: Within the groups, across the time periods of evaluation, there was a decline in cell survival in both the groups but was statistically significant in the MTA group. On interval-wise comparison, the decline in cell survival was statistically significant between the three groups at 72 h (P = 0.001).

Conclusions: E. officinalis preserved the vitality of the human primary dental pulp fibroblasts and has the potential to be developed into vital pulp therapy medicament.
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http://dx.doi.org/10.4103/JISPPD.JISPPD_85_18DOI Listing
November 2019

Antimicrobial susceptibility pattern of oral isolates of .

J Oral Maxillofac Pathol 2019 May-Aug;23(2):231-235

Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India.

Background: is involved in the etiology of localized aggressive periodontitis (LAP), a condition that frequently requires supplemental antibiotic therapy. Information on antimicrobial susceptibility pattern and guidelines for oral antibiotic therapy are not available on Indian patients.

Aim: The main aim of the present study was to screen clinical isolates on a panel of antibiotics commonly used for oral/systemic therapy.

Materials And Methods: The study included 40 strains of isolated from patients with LAP. The subgingival plaque was plated onto Trypticase Soy Serum Bacitracin Vancomycin Agar medium and incubated for 72 h, and suspected colonies were confirmed by phenotypic tests. Each isolate was tested against a panel of 12 antibiotics using MIC gradient strip test. ATCC strains of serotype A and C were used as standards. Performance and interpretation of the test were done according to the manufacturers' instructions. Distribution of MICs among isolates ( = 40) were used to calculate concentrations inhibiting 50% (MIC) and 90% (MIC) of strains.

Results: Moxifloxacin, cefotaxime and ceftriaxone showed excellent activity with 100% growth inhibition followed by amoxicillin, amoxiclav and doxycycline (>90% activity). The bacterial strains were moderately susceptible to cefuroxime, cefazolin and tetracycline but displayed poor susceptibility to clindamycin and azithromycin. All isolates were resistant to metronidazole.

Conclusion: The isolates of displayed a high level of resistance to azithromycin and clindamycin. Development of resistance against tetracycline also appears to be significant. Variable resistance among the different members of the cephalosporin group is a factor to be investigated further since susceptibility profile against these antibiotics and interpretative criteria for oral bacteria are not available.
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http://dx.doi.org/10.4103/jomfp.JOMFP_123_19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6714249PMC
September 2019

Effect of Resveratrol on biofilm formation and virulence factor gene expression of Porphyromonas gingivalis in periodontal disease.

APMIS 2019 Apr 12;127(4):187-195. Epub 2019 Mar 12.

Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences & Research Centre, Belgavi, Karnataka, India.

Periodontal disease is an oral inflammatory disease that destroys the tooth supporting periodontal tissues resulting in tooth loss. Porphyromonas gingivalis is a keystone pathogen that plays a significant role in periodontitis. In previous studies, resveratrol has shown significant results by targeting inflammatory and adhesive markers. Virulence factors of P. gingivalis play an important role in the bacterial adhesion and colonization. In this study, we aimed to demonstrate the anti-biofilm and anti-bacterial activity of resveratrol and also study the effect of resveratrol on the expression of virulence factor genes of P. gingivalis using reverse transcriptase polymerase chain reaction (RT-PCR). The anti-microbial and anti-biofilm activity of resveratrol on P. gingivalis was carried out by broth microdilution assay and biofilm adhesion reduction-crystal violet assay, respectively. We carried out the gene expression analysis by RT-PCR with the P. gingivalis treated compound to analyze the change in the expression of virulence factors: fimbriae and gingipain. Minimal inhibitory concentrations (MIC) of resveratrol against P. gingivalis and other clinical strains are in the range of 78.12-156.25 μg/mL. Resveratrol dose-dependently prevented the biofilm formation and also attenuated the virulence of P. gingivalis by reducing the expression of virulence factor genes such as fimbriae (type II and IV) and proteinases (kgp and rgpA). Resveratrol demonstrated superior anti-bacterial and anti-biofilm activity against P. gingivalis. There was significant reduction in the expression of fimbriae and gingipain with the resveratrol-treated compound. The results suggest that resveratrol, due to its multiple actions, may become a simple and inexpensive therapeutic strategy for treating periodontal disease.
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http://dx.doi.org/10.1111/apm.12930DOI Listing
April 2019

Detection of human papilloma virus-E6/E7 proteins of high-risk human papilloma virus in saliva and lesional tissue of oral squamous cell carcinoma patients using nested multiplex polymerase chain reaction: A comparative study.

J Oral Maxillofac Pathol 2018 Sep-Dec;22(3):318-324

General Dental Practitioner, Ferozepur, Punjab, India.

Introduction: Human papilloma virus (HPV)-associated oral squamous cell carcinoma (OSCC) shows different biological behavior as compared to tobacco-induced OSCC. Mere presence of HPV in OSCC is of no clinical significance; however, the integration of HPV-DNA through E6/E7 gene into the host genome is important as it affects the development and progression of OSCC.

Aim: The aim of this study was to determine the presence of E6/E7 proteins of high-risk (HR) HPV (HPV16 and HPV18) in saliva as well as lesional tissue of OSCC patients and to determine the use of saliva as an alternative to tissue for E6 and E7 proteins in OSCC.

Materials And Methods: Histopathologically confirmed 47 cases of OSCC were taken up for the study. The tumor tissue and saliva sample of each patient were obtained to detect the presence of HPV16 and HPV18 along with E6/E7 proteins in both samples by nested multiplex polymerase chain reaction (NMPCR). The data were analyzed using Student -test (2 tailed) and Wilcoxon signed-ranks test.

Results: In tumor tissue, 40.42% of cases showed HPV16 (19/47) positivity while 34.04% were HPV18 (16/47) positive; whereas, in salivary sample, 31.91% showed HPV16 (15/47) positivity while 25.53% of cases were HPV18 positive (12/47). Mean age of participants was 46.7 years, males showed no significant difference from females in the prevalence of HPV 16/18 with tongue being the most common site for the occurrence. There was no statistically significant difference for HPV16/18 presence in tissue and saliva sample of OSCC. Taking lesional tissue sample as standard, sensitivity and specificity for HPV16 and HPV18 in saliva by NMPCR was estimated at 68.42% and 92.86%, respectively. The accuracy level of NMPCR detection for HPV16 was 82.98% and HPV18 was 65.96%.

Conclusion: The study revealed no significant difference in the prevalence of HPV (16/18) among tissue and saliva of OSCC patients in Indian population. The study also found no difference in the level of DNA content of HPV in saliva and tissue indicating that saliva can be used as an alternative predictor of HPV positivity in OSCC.
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http://dx.doi.org/10.4103/jomfp.JOMFP_15_18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6306592PMC
January 2019

Study of microbial diversity in saliva and plaque samples from caries-free and caries-affected children using denaturing gradient gel electrophoresis.

J Indian Soc Pedod Prev Dent 2018 Oct-Dec;36(4):396-401

Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Maratha Mandal's Central Research Laboratory, Belgaum, Karnataka, India.

Background: Recent investigations have shown the possible involvement of bacteria other than mutans group and Lactobacilli in the etiology of caries. Molecular methods have been used to study the microbial diversity in caries-active (CA) and caries-free (CF) children. Among them, denaturing gradient gel electrophoresis (DGGE) is more popular and has been used in the present study.

Aims: The aim of the present study was to investigate the difference in bacterial diversity in saliva and plaque samples from CF and CA children using DGGE.

Materials And Methods: The study involved saliva and plaque samples from 56 children of which 28 were CF, 20 with CA, and 8 with white spot lesions (WSP). DNA was extracted and subjected to polymerase chain reaction amplification with universal primers. It was then run in polyacrylamide gel electrophoresis with gradients of urea and formamide and stained with SYBR green. Multiple bands were produced in each sample lane and each band represents one organism.

Statistical Analysis: A dendrogram was generated using Phoretix software and similarity index was calculated using a specific formula.

Results: Samples in each group formed several clusters indicating a specific pattern of the bacterial profile. Similarity coefficient was calculated based on the number of bands, intensity, and location. The diversity was less in the saliva and plaque samples of CA group as compared to those of CF and WSP groups.

Conclusions: DGGE can be used to study distinctive bacterial profiles in healthy and caries-affected sites. DGGE can be further developed as a pattern recognition tool with which to identify specific groups of bacteria. Saliva may be used to study bacterial diversity in dental caries.
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http://dx.doi.org/10.4103/JISPPD.JISPPD_206_18DOI Listing
May 2019

Detection of antibodies against Aggregatibacter actinomycetemcomitans in serum and saliva through ELISA in periodontally healthy individuals and individuals with chronic periodontitis.

Microb Pathog 2018 Dec 9;125:438-442. Epub 2018 Oct 9.

Department of Molecular Biology & Immunology, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belgaum, 590010, Karnataka, India.

Background: Periodontitis is a persistent polymicrobial infection, which leads to chronic inflammation in the tooth supporting tissues. Aggregatibacter actinomycetemcomitans are normal commensals of oral cavity but are low in number in periodontally healthy subjects. They are one of the major pathogens aetiologically linked to periodontal disease. Plasma and salivary antibody measurement may be useful to support diagnosis, disease activity, classification and prognosis of periodontitis. The aim of the present study was to investigate the association between the serum and salivary antibody levels to A. actinomycetemcomitans and therefore, to find whether this association was varying in different grades of periodontitis.

Method: Total of 50 periodontally healthy and 50 chronic periodontitis subjects (35-65 years) of both sexes were included for the study. 2 ml of un-stimulated saliva and 5 ml of venous blood was collected under sterile conditions. The detection of antibodies against A. actinomycetemcomitans in periodontally healthy individuals and individuals with chronic periodontitis was performed using indirect ELISA.

Results: Results showed serum IgG, IgA mean levels against A. actinomycetemcomitans were higher in chronic periodontitis subjects compared to mean levels in periodontally healthy subjects. Similarly, salivary IgG, IgA levels were also raised in chronic periodontitis patients as compared in healthy subjects. Also the mean levels of serum IgG and salivary IgA were increased as the severity of disease increased.

Conclusion: Antibody titer using saliva and serum could be useful tool for screening of patients with chronic periodontitis. Further, monitoring the various phases of treatment outcome using saliva could be a useful, non-invasive, prognostic indicator.
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http://dx.doi.org/10.1016/j.micpath.2018.10.007DOI Listing
December 2018

Comparative evaluation of the antimicrobial susceptibility and cytotoxicity of husk extract of and chlorhexidine as irrigating solutions against , and - An study.

J Indian Soc Pedod Prev Dent 2018 Apr-Jun;36(2):142-150

Department of Pedodontics and Preventive Dentistry, KLE University's KLE VK Institute of Dental Sciences, Belagavi, Karnataka, India.

Aim And Background: The aim of the present study is to evaluate and compare the antimicrobial susceptibility and cytotoxicity of Cocos nucifera and chlorhexidine (CHX) as irrigating solutions against Enterococcus faecalis, Prevotella intermedia, and Porphyromonas gingivalis.

Materials And Methods: The ethanolic extract of husk of C. nucifera was prepared. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined using the serial broth dilution method and its cytotoxicity was evaluated against human periodontal fibroblasts using 3-(4,5-dimethyl-thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. Antibacterial susceptibility for two irrigating solutions, namely 2% CHX gluconate irrigant (Group I) and 1.5% C. nucifera husk irrigant (Group II), was tested against P. gingivalis, P. intermedia, and E. faecalis.

Results: The MIC and MBC of C. nucifera husk extract for P. gingivalis were 468.75 μg/ml and 1562.5 μg/ml, for P. intermedia were 48.8 μg/ml and 1875 μg/ml, and for E. faecalis were 1562.5 μg/ml and 3750 μg/ml, respectively. The extract was nontoxic to the human periodontal fibroblast. Both the materials have shown similar antibacterial susceptibility and no difference was observed at baseline, 10, 30, and 60 min using two-way repeated measures of ANOVA. However, a statistically significant difference was observed between different time points for P. gingivalis and P. intermedia using Bonferroni multiple comparison test (f = 826.1390, P ≤ 0.05).

Conclusion: 1.5% of ethanolic husk extract of C. nucifera has a significant antibacterial action against polymicrobial dental biofilm and its activity is comparable to that of 2% CHX which validates its use as a future irrigating solution for overcoming bacterial resistance with synthetic agents.
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http://dx.doi.org/10.4103/JISPPD.JISPPD_1176_17DOI Listing
October 2018

Identification of multiple strains of using heteroduplex polymerase chain reaction in varying severity of chronic periodontitis.

Indian J Med Microbiol 2018 Jan-Mar;36(1):81-86

Department of Microbiology, SDM College of Medical Sciences and Hospital, Dharwad, Karnataka, India.

Aim: Research has demonstrated that there are multiple strains of Porphyromonas gingivalis with varying potency to cause periodontal disease. The current study aims at using heteroduplex polymerase chain reaction (PCR) to detect the strain diversity of P. gingivalis in periodontitis lesions of varying severity in a sample of the Indian population.

Materials And Methods: Subgingival plaque samples were collected from 60 individuals with varying severity of chronic periodontitis and 30 individuals with a clinically healthy periodontium. The samples were subjected to PCR analysis to identify P. gingivalis, followed by heteroduplex analysis to identify the strain diversity in a given sample. Bacterial culture was carried out as a comparative standard.

Results: Of the 56 samples that were positive for P. gingivalis by PCR, 54 samples yielded eight different heteroduplex patterns. Analysis of these patterns indicated that two strains of P. gingivalis were present in 41 individuals (45.6%) and three strains were present in 13 individuals (14.4%). Detection of P. gingivalis by PCR was significantly more in the periodontitis group as compared to the healthy group.

Conclusions: Species-specific PCR and heteroduplex analysis provide a simple and accurate method to analyse the strain diversity of P. gingivalis. P. gingivalis was detected in both healthy periodontal sites as well as sites with periodontitis. The presence of two or three P. gingivalis strains was seen in 60% of the samples.
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http://dx.doi.org/10.4103/ijmm.IJMM_17_434DOI Listing
September 2018

Antimicrobial activity of root canal irrigants against biofilm forming pathogens- An study.

J Conserv Dent 2017 May-Jun;20(3):147-151

Department of Conservative Dentistry and Endodontics, Maratha Mandal Dental College, Belgaum, Karnataka, India.

Aims: The aim of the study was to check the antimicrobial activity of the 5% Sodium hypochlorite, 2% Chlorhexidine, 0.10% Octenidine (OCT), and 2% Silver Zeolite (SZ) at different time intervals against a single species biofilm of , , and model prepared on a nitrocellulose membrane.

Settings And Design: nitrocellulose biofilm model was used to check antibacterial efficacy of root canal irrigants.

Materials And Methods: The nitrocellulose biofilm model was used to check the antibacterial activity of root canal irrigants. Single species biofilms were suspended into 96-well microtiter plate and treated with root canal irrigants for 1, 5, 10, 15, 30, and 60 s, respectively. The remaining microbial load in the form of colony-forming unit/ml after antimicrobial treatment was tabulated and data were statistically analyzed.

Statistical Analysis: SPSS version 17, Kruskal-Wallis ANOVA, Mann-Whitney U-test, and Wilcoxon matched pair test ( < 0.05) were used.

Results: All tested microorganisms were eliminated within 30 s by all the antimicrobial substances tested except normal saline. 2% chlorhexidine and 0.10% OCT were equally effective against at 30 s.

Conclusion: The newly tested irrigants have shown considerable antibacterial activity against selected single species biofilm. OCT (0.10%) can be used as an alternative endodontic irrigant.
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http://dx.doi.org/10.4103/JCD.JCD_38_16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5706312PMC
December 2017

Characterization and serotype distribution of Aggregatibacter actinomycetemcomitans: Relationship of serotypes to herpesvirus and periodontal status in Indian subjects.

Microb Pathog 2017 Sep 28;110:189-195. Epub 2017 Jun 28.

Department of Public Health Dentistry, Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences & Research Centre, Belagavi, Karnataka, India.

Background: The virulence of Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) in any individual depends on the type of strain of this bacterium. To our knowledge, there have been no studies reported in Indian subjects about A. actinomycetemcomitans serotype occurrence, co-existence with herpes virus and the possible influence of such co-existence on periodontal pathology.

Methods: Subjects for this study were a subset of a larger study to identify the prevalence of A. actinomycetemcomitans in chronic periodontitis. A total of 63 subjects (12 periodontally healthy and 51 with chronic periodontitis) who were positive for A. actinomycetemcomitans were serotyped for strain-level identification. The presence of Human Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) was tested in subgingival plaque samples by polymerase chain reaction.

Results: All five serotypes a to e were detected. Of the samples analyzed 38.09% harbored a single serotype, 36.5% had two serotypes, 6.3% demonstrated three and 4.7% demonstrated four serotypes. None of the samples showed presence of JP2 strain. Serotypes b, c, and e were most frequently identified in these individuals (46.03%, 36.5% and 38.09% respectively). Presence of serotypes b and c and absence of serotype d was associated with increased PD and CAL. Among 63 samples analyzed, 11 samples had CMV, four samples had EBV and nine samples had both these viruses. The PD and CAL were significantly higher (p = 0.04) when a combination of CMV and one of the serotypes was present indicating a pathological role of the coexistence.

Conclusion: Multiple serotypes are associated with chronic periodontitis in Indians, however, JP2 strains are not detectable in this cohort. Presence of multiple serotypes and a combination of any serotype with herpesvirus is associated with greater severity of the disease.
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http://dx.doi.org/10.1016/j.micpath.2017.06.041DOI Listing
September 2017

Antimicrobial Activity of Endodontic Medicaments and Vehicles using Agar Well Diffusion Method on Facultative and Obligate Anaerobes.

Int J Clin Pediatr Dent 2016 Oct-Dec;9(4):335-341. Epub 2016 Dec 5.

professor and Head, Department of Pedodontics and Preventive Dentistry, Maharishi Markandeshwar College of Dental Sciences and Research Ambala, Haryana, India.

Aims: The aim of this study was to determine the relative antimicrobial effectiveness of these endodontic medicaments and various vehicles using an agar well diffusion assay.

Materials And Methods: Double Antibiotic Paste(DAP), modified DAP, 2% Chlorhexidine gluconate and their combination with four vehicles namely Polyethylene glycol 400 (PEG), Propylene glycol (PG), combinations of PG with PEG and lastly Glycerine were tested using agar well diffusion assay. The minimum bactericidal concentration was noted against four standard strains of organisms ie ATCC( American Type Culture Collection) 25175, ATCC 12598, ATCC 35550 and ATCC 25922. Successful endodontic therapy depends upon thorough disinfection of root canals. In some refractory cases, routine endodontic therapy is not sufficient, so intracanal medicaments are used for proper disinfection of canals. Issues of resistance, limited spectrum of activity and lack of antifungal properties, the hunt for the ideal intracanal medicament continues. In this regard, the vehicles used to form the pastes play a supportive role by forming the appropriate consistency for placement and may dramatically influence their chemical characteristics like their solubility and diffusion. Thus, inorder to use safer and equally effective intracanal medicaments, Chlorhexidine gluconate is being unveiled in this study.

Results: The difference between the four vehicles when combined with the same endodontic medicament studied above is nonsignificant (NS) except against Porphyromonas gingivalis. Propylene glycol is significantly effective than Glycerine when used with DAP ie C+M medicament combination. (p = 0.029).

Conclusion: 2% chlorhexidine gluconate and modified DAP can definitely replace DAP and triple antibiotic paste as end-odontic medicaments with chlorhexidine having an added advantage of bactericidal action, substantivity, biocompatibility, low toxicity, and lesser chances of developing resistance.

How To Cite This Article: Nalawade TM, Bhat KG, Sogi S. Antimicrobial Activity of Endodontic Medicaments and Vehicles using Agar Well Diffusion Method on Facultative and Obligate Anaerobes. Int J Clin Pediatr Dent 2016;9(4):335-341.
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http://dx.doi.org/10.5005/jp-journals-10005-1388DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5233701PMC
December 2016

Indirect immunofluorescence technique to study expression of toll-like receptor 4 in chronic periodontitis.

Indian J Dent Res 2016 May-Jun;27(3):283-7

Department of Periodontics, Maratha Mandal's N.G.H. Institute of Dental Sciences and Research Centre, Belgaum, Karnataka, India.

Aim: The aim of this study was to analyze the expression level and localization of Toll-like receptor (TLR) 4 in gingival samples of healthy and chronic periodontitis subjects by indirect immunofluorescence technique (IFT).

Materials And Methods: In this study, gingival tissue samples were obtained from 25 healthy and 25 periodontitis individuals. The tissues were processed and the initial characterization was done by hematoxylin and eosin staining. The expression and localization of the TLR4 receptor were determined in the epithelial and connective layer cells of the gingival tissue using the indirect IFT. Immunofluorescence images were acquired and quantitative expression of TLRs was analyzed by calculating the percentage of cells showing positive results.

Results: We found that the healthy control group exhibited significantly lower values of TLR4 expression in comparison with the periodontitis patients. We also found that in patients with periodontitis the concentration of TLR4 was higher in the epithelium as compared to their expression in connective tissue cells.

Conclusions: These data suggested a definite involvement of TLR4 in initiating and progression of an inflammatory response in periodontitis.
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http://dx.doi.org/10.4103/0970-9290.186230DOI Listing
October 2017

Evaluation of antibacterial activity of Calotropis gigentica against Streptococcus mutans and Lactobacillus acidophilus: An in vitro comparative study.

J Conserv Dent 2015 Nov-Dec;18(6):457-60

Private Practitioner, Jaipur, Rajasthan, India.

Background: This study was conducted to evaluate in vitro antibacterial potential of ethanolic extract of Calotropis gigentica.

Materials And Methods: The inhibitory effect of the ethanolic extract was tested against Streptococcus mutans and Lactobacilli casei by using disc diffusion method.

Results: Ethanolic extract of Calotropis gigentica showed 16 mm and 14 mm of minimum inhibition zone at 1.25% concentration for S. mutans and lactobacilli, respectively.

Conclusion: Calotropis gigentica was found to effective against S. mutans and lactobacilli.
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http://dx.doi.org/10.4103/0972-0707.168809DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4693318PMC
January 2016

Synthesis, crystal studies, anti-tuberculosis and cytotoxic studies of 1-[(2E)-3-phenylprop-2-enoyl]-1H-benzimidazole derivatives.

Eur J Med Chem 2014 May 5;79:194-202. Epub 2014 Apr 5.

Department of Microbiology, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belgaum 590 010, India.

Series of 1-[(2E)-3-phenylprop-2-enoyl]-1H-benzimidazole derivatives were synthesized and characterized by spectral methods. Among 21 derivatives, single crystals of 3a and 3l were grown and their structural parameters were evaluated. Newly synthesized compounds were screened for anti-tubercular activity and the MIC was determined against Mycobacterium tuberculosis H37Rv by Microplate Alamar Blue Assay (MABA) method. Majority of the compounds exhibited a promising inhibition of M. tuberculosis and the molecules functionalized with electron-donating groups at C-2 carbon of benzimidazole moiety were found to be more active in inhibiting M. tuberculosis. Further, more promising compounds viz., 3b, 3i and 3l were tested for their cytotoxic activity. Compound 3l was found to display excellent activity (IC50 < 10 μg mL(-1)) with 100% cell lysis at 30 μg mL(-1) concentration against A549 (Human lung carcinoma) and 8E5 (Human; Acute Lymphoblastic Leukemia) cell lines.
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http://dx.doi.org/10.1016/j.ejmech.2014.04.017DOI Listing
May 2014

Analysis of Expression and Localization of TLR-2 by Immunofluorescent Technique in Healthy and Inflammed Oral Tissues.

J Clin Diagn Res 2013 Dec 15;7(12):2780-683. Epub 2013 Dec 15.

BDS, Private Practitioner, Vrisan Dental Care , Belgaum, India .

Introduction: Toll-like receptors (TLRs) are an important component of immune system. Among them, TLR-2 plays a dominant role in the oral tissues in initiating inflammation in chronic periodontitis. Not many studies have been done on quantitative expression of TLR-2 by using immunofluorescent techniques (IFT) in oral tissues. In this study, the expression and localisation of TLR-2 were detected in gingival tissues of chronic periodontitis patients and healthy individuals.

Material And Methods: Immuno Fluorescent Technique (IFT) was used for the expression and localization of TLR-2 in gingival tissue samples from 25 chronic periodontitis patients and from 25 healthy controls. Haematoxylin and Eosin staining was also done for all the samples to determine the histological characteristics of the gingival tissue samples.

Result: Both healthy and periodontitis gingival tissues expressed TLR-2. We found that the expression level of TLR-2 was higher in all the periodontitis patients than in healthy individuals. We also found out that the expression of TLR-2 was higher in the epithelial cells than in the connective tissue cells.

Conclusion: These data suggest a definite involvement of TLR-2 in initiating an inflammatory response in periodontal tissues. More studies are required to define the mechanisms and expression levels of TLR-2 in oral health and diseases.
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http://dx.doi.org/10.7860/JCDR/2013/6745.3745DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3919281PMC
December 2013

Estimation and comparison of salivary immunoglobulin A levels in tobacco chewers, tobacco smokers and normal subjects.

Oral Health Dent Manag 2013 Jun;12(2):105-11

Department of Oral Medicine and Radiology, NIMS University, Jaipur, Rajasthan, India.

Aims: To estimate the salivary immunoglobulin A (IgA) levels in tobacco chewers, tobacco smokers and normal subjects and to compare the salivary IgA levels among tobacco chewers and tobacco smokers.

Methods: The study group consisted of 80 subjects (tobacco users), 40 tobacco chewers and 40 tobacco smokers. Unstimulated whole saliva was collected from all tobacco users and 40 healthy age- and gender-matched non-tobacco users as control group. The study and control groups were divided into four subgroups based on age range. Salivary IgA levels were estimated by single radial immunodiffusion assay (SRID). All data were analysed using statistical software and to compare the results in three groups, single-factor analysis of variance was applied.

Results: The mean salivary IgA level in control group was 16.76 ± 1.37 mg/dl (SD); in tobacco chewers it was 7.89 ± 0.61 mg/dl (SD) and in tobacco smokers it was 6.55 ± 0.99 mg/dl (SD). The salivary IgA levels were decreased in tobacco chewers and tobacco smokers compared with the controls. Among the tobacco users, tobacco smokers had much reduced salivary IgA levels compared to tobacco chewers. All of these results were highly significant (P<0.001).

Conclusions: The present study showed that tobacco chewers and tobacco smokers had decreased salivary IgA levels and among tobacco users, tobacco smokers had much reduced salivary IgA levels compared to tobacco chewers in unstimulated whole saliva.
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June 2013

Microbiological analysis of oral samples for detection of Mycobacterium tuberculosis by nested polymerase chain reaction in tuberculosis patients with periodontitis.

Dent Res J (Isfahan) 2012 Nov;9(6):688-93

Department of Periodontics and Implantology, Kamineni Institute of Dental Sciences, Narketpally, Andhra Pradesh, India.

Background: Due to high prevalence of tuberculosis (TB) in India, presence of Mycobacterium tuberculosis in oral samples might influence the periodontal status of patients and also might pose a risk of transmission of TB during dental procedures through aerosols. Hence, this study aims to compare the periodontal status between TB and non-TB patients and to detect the presence of M. tuberculosis in plaque and saliva of TB and non-TB patients.

Materials And Methods: A total of 25 TB and 25 non-TB systemically healthy patients (age 21-49 years) were selected for this Clinico-Microbiological study. The oral hygiene and periodontal status of the patients were measured by using clinical indices and were compared using Mann-Whitney test (P < 0.05 = Significant). Pooled plaque and unstimulated salivary samples were collected and subjected to microbiological evaluation of M. tuberculosis by nested polymerase chain reaction. The detection rates were compared using Chi-square test (P < 0.001 = Highly significant).

Results: No significant difference was observed in periodontal clinical parameters measured between the groups. M. tuberculosis was detected in 92% of saliva and 68% of plaque samples of tuberculosis group, and even in 12% of saliva samples in nontuberculosis group patients.

Conclusion: The TB status of the patient did not influence the periodontal status. However, the presence of M. tuberculosis in plaque and saliva as detected in this study might pose a grave risk of transmission of the disease through aerosols during dental procedures.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3612214PMC
November 2012

Oral microbial carriage in oral squamous cell carcinoma patients at the time of diagnosis and during radiotherapy - a comparative study.

Oral Oncol 2012 Sep 17;48(9):881-6. Epub 2012 Apr 17.

Sri Dharmasthala Manjunatheshwara College of Dental Sciences and Hospital, Dharwad, Karnataka, India.

Objectives: Tobacco chewing habit, presence of squamous cell carcinoma in oral cavity and radiotherapy causes alterations in healthy oral microflora. Abnormal flora developed due to radiotherapy in oral squamous cell carcinoma (OSCC) patients can exacerbate mucositis and can cause systemic infections. The role of oral microorganisms in carcinogenesis is gaining interest recently. Abnormal flora in development of second tumor in the field of first tumor is to be established. The study fundamentally tries to evaluate the shift that occurs during the radiotherapy in OSCC patients.

Methods: Microbial analysis of saliva samples from OSCC patients undergoing radiotherapy, tobacco chewers and controls was undertaken. The microorganisms were grouped into categories as total aerobes, total anaerobes, candida, coliforms and gram negative anaerobic bacteria.

Results: The frequency of isolation of total aerobes, total anaerobes, coliforms and gram negative anaerobic bacteria was significantly high in OSCC patients compared to healthy controls whereas candida was isolated most frequently during radiation period. The tobacco chewers showed significant increase in colony forming units of total aerobes and coliforms. All the microbial groups were high in OSCC and radiotherapy patients. While OSCC patients showed significant increase in total anaerobes and gram negative anaerobes, candida was increased in radiotherapy patients only.

Conclusion: Habits promote coliforms. Tumor supports efficiently anaerobes and candida. The latter is supported more by radiation. The study stresses the importance on administration of appropriate antimicrobial therapy right at the time of diagnosis of the lesion.
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http://dx.doi.org/10.1016/j.oraloncology.2012.03.018DOI Listing
September 2012
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