Comparative evaluation of antimicrobial and antifungal efficacy of bioactive root-end filling materials: An in vitro study.

CONTEXT: Microorganisms are the main cause of failure of endodontic treatment. When retreatment fails periapical surgery followed by retrograde filling is done to seal the apex. A root-end filling material should have antimicrobial property as well as bioactive properties necessary for healing, repair, and regeneration of the apex. AIMS: The aim of this study was to evaluate the antibacterial and antifungal efficacy of three bioactive root-end filling materials: mineral trioxide aggregate (MTA) Plus, Biodentine, Endosequence root repair material (ERRM) against Enterococcus faecalis and Candida albicans. SUBJECTS AND METHODS: E. faecalis and C. albicans standard bacterial strains were used. 100 µl was taken from liquid cultures of E. faecalis and planted in Mueller-Hinton agar and the same amount of C. albicans was planted in Sabouraud dextrose agar by lawn culture. MTA Plus, Biodentine, and ERRM were aseptically filled into the opened pits. Following this, the media were kept in the drying oven at 37°C for 24, 48, and 72 h and the diameters of the inhibition zones were measured. STATISTICAL ANALYSIS USED: Statistical analysis was carried out by Kruskal-Wallis, Post hoc (Mann-Whitney), Friedman, and Post hoc (Wilcoxon-sign) test. RESULTS: Among the three groups, the antimicrobial activity of Biodentine against E. faecalis was statistically higher than MTA Plus and ERRM (P < 0.05). Antifungal activity of MTA Plus against C. albicans was statistically higher than Biodentine and ERRM (P < 0.05). ERRM showed the smallest inhibition zone against E. faecalis and C. albicans among the three groups (P < 0.05). CONCLUSIONS: Biodentine exhibited the greatest antimicrobial activity and MTA Plus exhibited the greatest antifungal activity among the three groups. ERRM exhibited the least antibacterial and antifungal activity among the three groups.

Estimation and correlation of serum and salivary glucose and immunoglobulin A levels and salivary candidal carriage in diabetic and non-diabetic patients.

Background. A correlation has been noted between diabetes mellitus (DM) and changes in the oral cavity. The present study aimed to estimate, compare, and correlate serum and salivary glucose and IgA levels and salivary candidal carriage in diabetic and non-diabetic individuals. Methods. Eighty-eight subjects were categorized into three groups: group 1 (controlled DM; n=27), group 2 (uncontrolled DM; n=32) and group 3 (non-diabetics; n=29). Serum and salivary glucose levels were estimated by glucose oxidase/peroxidase method, serum and salivary IgA by a diagnostic kit, and candidal colonization by inoculating samples into Sabouraud dextrose agar plate. Statistical analyses were carried out by one-way ANOVA, post hoc Tukey tests, and Pearson's correlation coefficient. Results. Significant elevation of serum IgA levels was observed in group 2 compared to group 3 and significant decreases in salivary IgA levels in groups 1 and 2. The candidal carriage was significantly higher in group 2 compared to group 3. Serum glucose and salivary IgA levels showed a significant correlation in group 1. There was a positive correlation between serum/ salivary glucose and serum/salivary IgA levels in group 2. In addition, there was a significant correlation between serum glucose and serum IgA levels in group 3. Conclusion. Saliva could be a potential, non-invasive diagnostic tool to estimate glucose levels. The evaluation of salivary components, like IgA, might be useful in diagnosing and managing oral manifestations in diabetic individuals. Elevated salivary glucose levels contribute to elevated candidal carriage, making individuals susceptible to oral candidiasis.

A novel ready-to-use dry-reagent polymerase chain reaction for detection of Escherichia coli & Shigella species.

Background & objectives: Polymerase chain reaction (PCR) has wide acceptance for rapid identification of pathogens and also for diagnosis of infectious conditions. However, because of economic and expertise constraints, a majority of small or peripheral laboratories do not use PCR. The objective of the present study was to develop a dry-reagent PCR assay as an alternative to conventional PCR to assess its applicability in routine laboratory practice using malB gene for identification of Escherichia coli as a model. Methods: A total of 184 isolates were selected for the study comprising clinical isolates of E. coli and non-E. coli including Shigella sp. and a few other control strains. The DNA was isolated from all the isolates. The isolated DNA as well as the overnight grown bacterial cultures were subjected to both conventional wet PCR and dry-reagent PCR. Results: The genomic DNA isolated from E. coli showed amplification of malB gene in both conventional wet and dry-reagent PCR and the band was observed at 491 bp. In dry-reagent PCR, the overnight grown E. coli cells also showed positive result. The non-E. coli strains other than Shigella sp. showed negative in both conventional wet and dry-reagent PCR. Shigella sp. showed positive in both conventional wet and dry-reagent PCR. Interpretation & conclusions: Considering the elimination of genomic DNA isolation step, and similar results with the conventional wet PCR, dry-reagent PCR may be a good alternative for the conventional wet PCR.

Bacterial Biofilms in Chronic Rhinosinusitis and Their Implications for Clinical Management.

To study the microbiological profile in patients with chronic rhino-sinusitis. To correlate disease severity with the presence of biofilms and host risk factors. To assess outcome of Sinus Surgery 2 weeks post operatively in terms of presence of bacteria and their ability to form biofilm. Prospective study. 50 cases of chronic rhino-sinusitis requiring Functional Endoscopic Sinus Surgery admitted in SDM Hospital, Dharwad, Karnataka were studied using intra-operative mucosal samples for microbiological analysis. The organisms isolated were tested for biofilm forming ability using three in vitro tests. Severity of disease was assessed using SNOT 22 scoring system. Of 50 cases studied, 66% showed presence of chronic rhino-sinusitis with polyposis and had higher SNOT scores compared to those without polyps. Bacterial isolates were obtained from only 17 samples. Staphylococcus species was isolated from 16 samples and Klebsiella pneumoniae from one. 11 Staph spp. isolates showed biofilm forming ability in vitro. Postoperative events in 3 cases yielded biofilm-forming Staphylococcus. Staphylococcus was the most dominant organism isolated and 11 isolates were biofilm formers. Thus the detection of biofilm forming organisms can be considered as a negative prognostic indicator and should forewarn the surgeon about the risk of recurrence.

Development of a dry-reagent mix-based polymerase chain reaction as a novel tool for the identification of Acinetobacter species and its comparison with conventional polymerase chain reaction.

BACKGROUND: Nosocomial infections are often caused by multidrug-resistant bacteria and the incidence is increasing. Acinetobacter, a Gram-negative bacillus, is commonly associated with the use of intravascular catheterization and airway intubation. Polymerase chain reaction (PCR) for identification of Acinetobacter baumannii from samples has been standardized that use conventional wet-reagent mix. We have designed and optimized a dry-reagent mix for identification of Acinetobacter species by PCR. The dry-reagent mix can be stored at room temperature, has less chances of contamination, and thus can be used at point-of-care diagnosis. AIM AND OBJECTIVE: The present work was focused on comparing the sensitivity and specificity of dry-reagent PCR mix over conventional wet-reagent PCR mix for identification of Acinetobacter species. MATERIALS AND METHODS: Conventional wet-reagent mix based and dry-reagent mix based PCR were carried out for the DNA isolated from Acinetobacter species. The latter was also applied directly on bacterial growth without prior DNA extraction process. Equal numbers of bacterial isolates other than Acinetobacter species were also subjected to identification by the same protocols for determining the sensitivity and specificity of the test. RESULTS: The Acinetobacter species showed amplification of the target rpoB gene and the band was observed at 397 bp. The dry-reagent PCR mix results matched completely with the conventional wet-reagent PCR mix assay. All the non-Acinetobacter isolates were negative for the PCR. This indicates that the test is highly specific. The dry-reagent mix also contained an enzyme resistant to PCR inhibitors and capable of amplifying DNA directly from cells. CONCLUSION: Performance of dry-reagent PCR mix without the need for DNA extraction and preparation of a PCR mix proved to be more sensitive and reduce the handling error, minimizes the time, manual work, and skilled labor.

Systemic Cytokines in Type 2 Diabetes Mellitus and Chronic Periodontitis.

BACKGROUND: Cytokine dysregulation plays an important role in Type 2 Diabetes Mellitus (T2DM) and Chronic Periodontitis (CP) with a commonality in pathogenic mechanisms. CP is considered the sixth complication of diabetes and may have an increased influence on systemic levels of cytokines in individuals with T2DM. OBJECTIVE: This study investigated two pro-, and two presumed anti-inflammatory cytokines and their ratios in the serum of healthy individuals, in chronic periodontitis with and without T2DM with, and without CP and in CP alone aimed at evaluating the systemic inflammatory burden of a local oral infection. METHODS: Eighty participants were divided equally into four groups as healthy volunteers (H) and patients having T2DM with, and without CP (T2DM+CP, and T2DM) and only CP (CP). Serum samples were collected to measure glycated hemoglobin (HbA1c), Random Blood Sugar (RBS) and also Tumor Necrosis Factor (TNF)-α, Interleukin (IL)-4, -6 - 10 were assessed using commercially available ELISA kits. RESULTS: The cytokines were detected in all groups. Significant differences were observed between groups for all the clinical, biochemical parameters and cytokines. Cytokine levels and the ratios showed significant correlations. The ratios of the cytokines differed significantly amongst groups, were highest in T2DM+CP. CONCLUSION: In this study, the cytokine ratios provided a qualitative profile along with the absolute levels in T2DM with periodontitis, indicative of an intensified systemic inflammatory state.

Identification of genus Acinetobacter: Standardization of in-house PCR and its comparison with conventional phenotypic methods.

BACKGROUND: Acinetobacter is grouped under nonfermenting Gram-negative bacilli. It is increasingly isolated from pathological samples. The ability of this genus to acquire drug resistance and spread in the hospital settings is posing a grave problem in healthcare. Specific treatment protocols are advocated for Acinetobacter infections. Hence, rapid identification and drug susceptibility profiling are critical in the management of these infections. AIMS: To standardize an in-house polymerase chain reaction (PCR) for identification of genus Acinetobacter and to compare PCR with two protocols for its phenotypic identification. METHODOLOGY: A total of 96 clinical isolates of Acinetobacter were included in the study. An in-house PCR for genus level identification of Acinetobacter was standardized. All the isolates were phenotypically identified by two protocols. The results of PCR and phenotypic identification protocols were compared. RESULTS: The in-house PCR standardized was highly sensitive and specific for the genus Acinetobacter. There was 100% agreement between the phenotypic and molecular identification of the genus. The preliminary identification tests routinely used in clinical laboratories were also in complete agreement with phenotypic and molecular identification. CONCLUSION: The in-house PCR for genus level identification is specific and sensitive. However, it may not be essential for routine identification as the preliminary phenotypic identification tests used in the clinical laboratory reliably identify the genus Acinetobacter.

Global eradication of measles: Are we poised?

Measles, a highly infectious viral disease is the next target for eradication following poliovirus. Decades of experience with highly effective vaccination has invigorated us to take on this virus. The task is not only Titanic but is laced with intricate issues. Recently, an outbreak of fever with rash occurred on a tertiary care teaching hospital campus and was confirmed serologically as measles outbreak by IgMELISA. Therefore, we searched the literature related to outbreaks, transmission of the measles virus, age groups involved, vaccination strategies, vaccination failure and epidemiological features of the disease and reviewed the possible reasons for such outbreaks and problems in the global eradication of the virus.

Cytokine ratios in chronic periodontitis and type 2 diabetes mellitus.

AIM: Chronic periodontitis may influence systemic cytokines in type 2 diabetes. This study aimed to evaluate the cytokine ratios in type 2 diabetes with, and without chronic periodontitis. MATERIALS & METHODS: Gingival status, periodontal, glycemic parameters and serum cytokines were evaluated in participants grouped as healthy, chronic periodontitis, and type 2 diabetes with, and without chronic periodontitis. RESULTS AND CONCLUSIONS: Cytokine ratios showed significant differences in type 2 diabetes and chronic periodontitis, were highest in participants having both type 2 diabetes and chronic periodontitis, with a statistically significant cut-off point and area under curve by receiver operating characteristic.

Evaluation of minimal inhibitory concentration of two new materials using tube dilution method: An in vitro study.

AIM: The aim of the study is to evaluate and compare the antimicrobial efficacy of two new materials MTA Plus and Biodentine with ProRoot MTA using tube dilution method. MATERIALS AND METHODS: The materials used were ProRoot MTA (Dentsply), MTA Plus (compounded by Prevest Denpro, Jammu, India for Avalon Biomed Inc, USA) and a calcium silicate based material Biodentine (Septodont, Saint-Maur-des-Fosses, France). Doubling dilutions of the material were prepared in Sabouraud's dextrose broth (SDB) and Brain Heart Infusion (BHI) broth for Candida albicans and Enterococcus faecalis, respectively. The minimal concentration at which inhibition of microorganism occurred was measured and noted as minimal inhibitory concentration (MIC) of the material. RESULTS: There was no statistically significant difference between the materials against C. albicans. Biodentine was statistically significant than MTA Plus against E. faecalis (P-value-0.022). ProRoot MTA was statistically significant at different time intervals against E. faecalis (P-value-0.001). CONCLUSION: ProRoot MTA and Biodentine proved to have antimicrobial property. MTA Plusproved as a good antifungal agent.

PCR as a diagnostic tool for extra-pulmonary tuberculosis.

INTRODUCTION: Extra-Pulmonary Tuberculosis (EPTB) accounts for approximately 40% of the tuberculosis cases. Though it is not communicable, it is a significant cause of morbidity. This study was conducted to know the efficacy of the Polymerase Chain Reaction (PCR) as an additional tool, along with the conventional methods, in the diagnosis of EPTB. MATERIAL AND METHODS: Clinical samples were collected from suspected cases of EPTB. The Ziehl-Neelsen staining (ZNS), culture on the Lowenstein-Jensen medium (LJM) and PCR testing with the use of a commercial kit were performed on the homogenized samples. RESULTS: A total of 182 samples which were received for the molecular diagnosis of EPTB were also tested by ZNS and culture on LJM for the presence of Mycobacterium tuberculosis. Of these, 22 were positive by at least one of the tests which were used. PCR detected the maximum number of cases of EPTB, followed by culture. The results of PCR and the conventional tests were analyzed by using McNemar's test for the correlated proportions-the exact method of 'IBM SPSS Statistics 20'. The analysis showed a statistical significance. CONCLUSIONS: Whenever they are feasible, using all the available tests in combination increases the laboratory detection rates of M. tuberculosis from clinical samples. PCR must be included in the diagnostic panel of EPTB.

Phenotypic detection of inducible clindamycin resistance among Staphylococcus aureus isolates by using the lower limit of recommended inter-disk distance.

CONTEXT: Clindamycin is one of the important alternative antibiotics in the therapy of Staphylococcus aureus, particularly in methicillin-resistant S. aureus (MRSA) infections. Inducible clindamycin resistance (iMLS B--inducible Macrolide-Lincosamide-Streptogramin B resistance) is a critical factor in antimicrobial susceptibility testing. AIMS: To know the rate of inducible clindamycin resistance among clinical isolates of Staphylococcus aureus in our hospital by Disk approximation test (D-test) using the average recommended inter-disk distance and comparing the results with that of D-test using the lower limit of recommended inter-disk distance. MATERIALS AND METHODS: A total of 51 erythromycin-resistant and clindamycin-susceptible S. aureus isolates were subjected to disk approximation testing with 21 +/- 1 mm and 15 mm edge-to-edge distance between the clindamycin and erythromycin disks. STATISTICAL METHODS: Z-test levels. RESULTS: Among 51 erythromycin-resistant and clindamycin-susceptible S. aureus isolates, 25 (49%) were recorded as inducible clindamycin resistant by D-test with 21 +/- 1 mm edge-to-edge distance between the clindamycin and erythromycin disks. When we re-tested all the 51 strains by D-test with 15 mm inter-disk distance, we identified 14% more iMLS B strains previously reported as D-test negative. Z-test for MRSA indicates that 15 mm edge-to-edge distance has significant advantage. CONCLUSIONS: Since the incidence of inducible clindamycin resistance is high (63% in our study), accurate identification of inducible clindamycin resistance is important to prevent therapeutic failure in infections caused by these strains. We suggest the use of D-test with 15 mm edge-to-edge inter-disk distance for detecting iMLS B .