Analysis of distribution and localization of proteins of the reelin signalling pathway in mesial temporal lobe epilepsy.

INTRODUCTION: Granule cell dispersion (GCD) is pathognomonic of hippocampal sclerosis seen in the mesial temporal lobe epilepsy (MTLE). Current animal studies indicate deficiency of Reelin is associated with abnormal granule cell migration leading to GCD. The present study aimed to evaluate complete Reelin signalling pathway to assess whether Reelin deficiency is related to MTLE. MATERIALS AND METHODS: Hippocampal sclerosis was confirmed by H and E stain. To explore the amount and cellular location of the Reelin cascade molecules, the hippocampal tissues from MTLE surgery and controls (n = 15 each) were studied using Immuno-histochemistry (IHC). Additionally, confocal imaging was used to validate the IHC findings by co-localization of different proteins. Quantification of IHC images was performed using histo-score and confocal images by Image J software. RESULTS: Immune expression of active Reelin was significantly reduced in patients. Reelin receptors were deranged, apolipoprotein E receptor 2 was increased while very low-density lipoprotein receptor was reduced. Disabled-1, a downstream molecule was significantly reduced in MTLE. Its ultimate target, cofilin was thus disinhibited and expressed more in MTLE. Reelin cleaving protease, matrix metalloprotease-9 (MMP-9) and MMP-9 inhibitor, tissue inhibitor of matrix protease-1, showed reduced expression in extracellular matrix. Semi-quantification of immunohistochemistry was done using Histo (H) score. H score of Reelin in diseased patients was 15 against 125 for control patients. These results were validated by confocal fluorescence microscopy. CONCLUSIONS: Reelin signalling cascade was deranged in chronic MTLE. Pharmacological manipulation of Reelin cascade can be done at various levels and it may provide novel treatment options for MTLE.

Effectiveness of subgingival irrigation and powered toothbrush as home care maintenance protocol in type 2 diabetic patients with active periodontal disease: A 4-month randomized controlled trial.

Background: There is a bidirectional link between diabetes and periodontal disease. Control of active periodontal infection is an essential requisite to maintain optimal oral and systemic health in diabetic patients. The purpose of this study was to evaluate the efficacy of subgingival irrigation and powered toothbrush as home care maintenance protocol in type 2 diabetic patients with active periodontal disease compared to routine oral hygiene. Materials and Methods: Forty (n = 40) diabetic (HbA1c >7%) patients were enrolled in this parallel, examiner-blind, interventional clinical trial. Patients were randomized into two groups: Group A (sonic toothbrush and irrigation with water twice daily) or Group B (manual toothbrush and mouth rinsing with 0.12% Chlorhexidine gluconate (CHX) twice daily)). All patients received Phase I therapybefore the start of the study. Clinical parameters (plaque index [PI], gingival index [GI], oral hygiene index [OHI], pocket depth [PD], clinical attachment level [CAL], and bleeding index [BI]) were assessed at baseline, 1, 2, and 4 months. Levels of C-reactive protein (CRP), HbA1c, and interleukin (IL)-1 ß were assessed at baseline and 4 months only. Verbal and written instructions were provided to each subject specific to their intervention allocation. Descriptive, parametric, and nonparametric analyses were used where appropriate. Results: Sixteen (n = 16) patients in Group A and fifteen (n = 15) patients in Group B completed the 4-month study. Both groups showed a significant difference in BI, PD, CAL, and HbA1c from baseline to 4 months. There were no differences within groups for OHI, GI, or PI and CRP, IL-1 ß. The results are based on an underpowered study due to the drop out of 9 patients reducing the number below the needed 19 patients per group based on the power analysis. Conclusion: Results from this study provide information for future studies on self-care regimens for individuals living with Type 2 diabetes.

Influence of periodontitis and diabetes on structure and cytokine content of platelet-rich fibrin.

BACKGROUND: Platelet-rich fibrin (PRF) is a second-generation platelet concentrate with multiple applications in wound healing and regeneration in both periodontitis and diabetes. However, the three dimensional (3-D) structure and cytokine content of PRF might be altered in patients suffering from either/both of the chronic inflammatory conditions, ultimately influencing the efficacy of PRF as a biomaterial for regenerative medicine. AIM: The aim of the present study was hence to evaluate the effect of both these chronic inflammatory diseases on the 3-D structure of PRF membrane. An attempt was also made to compare the growth factor content between the plasma and RBC ends of the prepared PRF gel. MATERIALS & METHODS: L-PRF was prepared for twenty participants, healthy (5), periodontitis (5), T2DM (5) and T2DM with periodontitis (5). Porosity and fiber diameter of PRF membranes was visualized under FE-SEM and measured using ImageJ Software. PDGF-BB and TGF-ß1 levels in PRF gel were assessed by ELISA. RESULTS: The average diameter of fibrin fibers under FE-SEM was 0.15 to 0.30 micrometers. Porosity was higher at the plasma end (p = 0.042). Red blood cell (RBC) end of the membrane had thinner fibers arranged in a comparatively more dense and compact structure with smaller porosities. Healthy subjects had the least porous PRF compared to subjects with either/both of the chronic conditions. PDGF-BB levels were similar along all the four groups. TGF-ß1 levels were highest in healthy subjects. DISCUSSION: 3-D structure and growth factor content of PRF are influenced by a person's periodontal and/or diabetic status. The RBC end of the PRF membrane, as compared to the plasma end, has thinner fibers arranged in a comparatively more dense and compact structure with smaller porosities, and hence should be favored during periodontal regenerative procedures. CONCLUSION: Both periodontitis and diabetes have a significant influence on the 3-D structure and growth factor content of PRF produced.

Expressional Study of Permeability Glycoprotein and Multidrug Resistance Protein 1 in Drug-resistant Mesial Temporal Lobe Epilepsy.

Introduction: About 30% of patients with epilepsy do not respond to anti-epileptic drugs, leading to refractory seizures. The pathogenesis of drug-resistance in mesial temporal lobe epilepsy (MTLE) is not completely understood. Increased activity of drug-efflux transporters might be involved, resulting in subclinical concentrations of the drug at the target site. The major drug-efflux transporters are permeability glycoprotein (P-gp) and multidrug-resistance associated protein-1 (MRP-1). The major drawback so far is the expressional analysis of transporters in equal numbers of drug-resistant epileptic tissue and age-matched non-epileptic tissue. Methods: We have studied P-gp and MRP-1 drug-efflux transporters in the sclerotic hippocampal tissues resected from the epilepsy surgery patients (n=15) and compared their expression profile with the tissues resected from non-epileptic autopsy cases (n=15). Results: Statistically significant over expression of both P-gp (P<0.0001) and MRP-1 (P=0.01) at gene and protein levels were found in the MTLE cases. The fold change of P-gp was more pronounced than MRP-1. Immunohistochemistry of the patient group showed increased immunoreactivity of P-gp at blood-brain barrier and increased reactivity of MRP-1 in the parenchyma. The results were confirmed by confocal immunofluorescence microscopy. Conclusion: Our results suggested that P-gp in association with MRP-1 might be responsible for the multi-drug resistance in epilepsy. P-gp and MRP-1 could be important determinants of bio availability and tissue distribution of anti-epileptic drugs in the brain which can pharmacologically inhibited to achieve optimal drug penetration to target site.

Psoriasis and Periodontitis: Exploring an association or lack thereof.

OBJECTIVES: Psoriasis is a common, chronic, non-communicable skin disease with no clear etiology or cure. Periodontitis is a chronic inflammatory condition which is now known to significantly influence various systemic diseases as an established risk factor. This study aimed at comparatively evaluating the periodontal status of Psoriatic patients vis. a vis. that of age and gender matched systemically healthy volunteers. An attempt was also made to explore a possible association, if any, amongst the two diseases. MATERIALS AND METHODS: Forty two residents of Chandigarh, suffering from Psoriasis and attending the Psoriasis Clinic of Department of Dermatology & Venereology, Post Graduate Institute of Medical Education & Research, Chandigarh were recruited over a period of 10 months (Case group) and their periodontal status was compared with forty two age and gender matched systemically healthy volunteers (Control group) randomly selected from the Out Patient Department of Periodontics, Dr. Harvansh Singh Judge Institute of Dental Sciences & Hospital, Panjab University, India. Their serum IL-33 levels were evaluated and compared in an attempt to identify an underlying common pathological pathway. RESULTS: The periodontal status was comparable in the two groups in terms of the debris index (p = 0.932), calculus index (p = 0.088), plaque index (p = 0.097), and mean clinical attachment loss (p = 0.401). A higher bleeding points index was recorded amongst healthy individuals as compared to the Psoriasis group, the difference being statistically significant (p = 0.001). The mean number of teeth were more in the Psoriasis group as compared to the healthy group (p=0.034). IL 33 levels were also not significantly different (p = 0.491). CONCLUSION: Contrary to currently available evidence in literature, the study did not find a statistically significant association between Psoriasis and Inflammatory Periodontal Disease.

Impact of concurrent diabetes on periodontal health in patients with acromegaly.

Previous studies have suggested excess GH/IGF1 secretion in patients with acromegaly is protective for periodontal health. Diabetes is prevalent comorbidity in patients of acromegaly and is associated with worsening of periodontal disease. The present study evaluates the periodontal health and cytokines status in treatment-naive active acromegaly patients with and without diabetes. Eleven patients, each of acromegaly with and without diabetes and 20 healthy controls were enrolled. Periodontal parameters were assessed. GCF and blood samples for IL-6, TGF-ß1, and PDGF were obtained. Serum GH, IGF1, HbA1c, pituitary hormones and MRI sella were performed in patients with acromegaly. There was no significant difference in periodontal status of patients with acromegaly and healthy controls. However, a significant increase in serum IL-6 (p = 0.019) and TGF-ß1 (p = 0.025) levels in patients with acromegaly was observed and all patients had concurrent hypogonadism. Nevertheless, the patients with acromegaly having diabetes had modestly higher CAL and PD and serum IL-6 levels (p = 0.051), but it could not exert adverse effects on periodontal health in presence of GH/IGF1 excess. GH/IGF1 excess did not exert a protective effect on periodontal status in acromegaly, possibly due to concurrent hypogonadism and opposing cytokines; however, it could mask the ill-effects of diabetes on periodontal health.

Evaluation of Curcumin Gel as Adjunct to Scaling & Root Planing in Management of Periodontitis- Randomized Clinical & Biochemical Investigation.

BACKGROUND: Successful management of periodontitis requires treatment strategy that integrates therapies addressing both pathogen and host aspects of disease etiology. To evaluate sub gingivally applied curcumin gel in treatment of chronic periodontitis based on clinical and biochemical parameters. MATERIALS & METHODS: A randomized, double blind, parallel-group trial was carried out on 30 patients suffering from chronic generalized periodontitis with probing pocket depth≥5mm on at least 4 sites who were then randomly allocated to two groups. Control group was treated with Scaling and Root Planing (SRP) alone while experimental group was treated with SRP followed by subgingival application of curcumin gel. Saliva collection was done and the clinical parameters were recorded at baseline and follow up periods. Saliva analysis for IL-1ß was done by ELISA. The statistical differences for the intra-group and intergroup measurements were analyzed by using Mann Whitney test. Spearman's rank correlation coefficient was used to examine the relationship between Interleukin - 1ß and clinical parameters. RESULTS: Study elucidated mild adjunctive benefit of curcumin gel in reduction of gingival inflammation for a limited period of time. Though improvement in other clinical parameters was also greater in subjects treated with curcumin gel, results were not statistically different from those treated with SRP alone. None of the subjects who received curcumin gel experienced any adverse effect. CONCLUSION: Within limitations, it can be concluded that single application of curcumin (turmeric) gel has limited added benefit over scaling and root planing in treatment of chronic periodontitis.

Salivary IgA, Interleukin-1ß and MMP-8 as Salivary Biomarkers in Chronic Periodontitis Patients.

OBJECTIVE: To compare the clinical parameters and levels of salivary immunoglobulin A (IgA), Interleukin-1ß (IL-1ß) and matrix metalloproteinase-8 (MMP-8) in patients with moderate to severe chronic periodontitis and in individuals with healthy gingiva. METHODS: Levels of clinical parameters plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment level (CAL) and bleeding on probing (BOP) along with salivary biomarkers salivary IgA, IL-1ß and MMP-8 were recorded among 50 adults (30 test subjects with moderate to severe generalised chronic periodontitis, constituting group A, and 20 periodontally healthy controls - group B). Clinical evaluation was done before oral prophylaxis, and 6 weeks and 12 weeks after oral prophylaxis, and saliva samples were obtained before and 12 weeks after oral prophylaxis. Salivary IgA, IL-1ß and MMP-8 levels in saliva were assessed using enzyme-linked immunosorbent assay. RESULTS: In group A, there were highly significant differences in terms of PI, GI, PD, CAL and BOP before oral prophylaxis, and 6 weeks and 12 weeks after oral prophylaxis when compared at these intervals. Differences in their levels in group B were non-significant at such intervals except PI. Mean levels of salivary IgA, IL-1ß and MMP-8 in chronic periodontitis patients at baseline were significantly higher than in the periodontally healthy group. Their levels in group A decreased significantly 12 weeks after oral prophylaxis, but remained static in group B. CONCLUSION: The levels of salivary IgA, IL-1ß and MMP-8 showed significant reduction after oral prophylaxis, suggesting that these biomarkers could facilitate the screening, early diagnosis, and management of periodontal disease.

Role of cardiovascular disease markers in periodontal infection: Understanding the risk.

CONTEXT: Both periodontitis and cardiovascular diseases (CVD) represent chronic inflammatory conditions, and periodontal infections have been postulated to perpetuate the progression of CVD's. However, limited evidence is available to prove the causal relationship. AIM: An effort in exploring this interrelation has been made in this study. The role of two inflammatory mediators, soluble CD40 ligand (sCD40 L) and monocyte chemoattractant protein-1 (MCP-1) has been established in progression and acute precipitation of CVD's. Due to a close link between these two mediators, the present study was designed to correlate the levels of sCD40 L and MCP-1 in serum and gingival crevicular fluid (GCF) of patients with chronic periodontitis. METHODS: Fifteen healthy and 30 patients of severe chronic periodontitis (diseased) participated in the study. Patients of the diseased group underwent scaling/root planning. The evaluation of plaque index, gingival index, probing depth, clinical attachment level, and a collection of serum and GCF samples was done at baseline and 6 weeks following periodontal therapy. The sCD40 L and MCP-1 levels were quantified using ELISA. RESULTS: The sCD40 L levels correlated strongly with MCP-1 levels in both GCF (r = 0.888) and serum (r = 0.861) in patients of chronic periodontitis. The relationship between the levels of the two markers was maintained in GCF (r = 0.868) and serum (r = 0.750) after Phase I periodontal therapy. CONCLUSIONS: The positive correlation observed suggests this pathway as one of the mechanisms that may lead to increasing severity of periodontal disease and its systemic effects. Further research efforts should be made in designing appropriate clinical trials, starting at an early stage and monitoring the potential benefits of maintenance of oral hygiene on cardiovascular health.

Soluble CD40 ligand: a novel biomarker in the pathogenesis of periodontal disease.

OBJECTIVES: Periodontitis involves a complex interplay of micro-organisms and host immune response via numerous mediator molecules playing strategic roles in its pathogenesis. Soluble CD40L (sCD40L) is one such co-stimulatory molecule which is essential for T-helper cell activation and is a well-known risk indicator of cardiovascular diseases. The levels of this marker in crevicular fluid of patients of chronic periodontitis have been explored in the present study for the first time along with an analysis of its association with levels in serum in otherwise systemically healthy patients. METHODOLOGY: Sixty patients 20 healthy and 40 of chronic periodontitis (18 moderate and 22 severe) participated in the study. Patients of the diseased group underwent non-surgical periodontal therapy. Clinical evaluation and collection of gingival crevicular fluid (GCF) and serum samples was done at baseline, and 6 weeks after phase I periodontal therapy. sCD40L levels were quantified in the fluids using ELISA. RESULTS: Levels of sCD40L in GCF were significantly higher in the diseased group (p ≤ 0.001) and strongly correlated not only with increasing severity of disease but also with levels in serum. In post-treatment, the levels decreased significantly in both the biological fluids (p ≤ 0.001). CONCLUSIONS: The present study brings to light the role of sCD40L as a novel marker in mediating periodontal destruction and disease progression. Evaluation of local treatment outcomes seems promising in minimizing these effects. CLINICAL RELEVANCE: Positive association of its local levels with those in serum further implicates the possibility of widespread systemic effects of this infection.

Role of monocyte chemoattractant protein-1 (MCP-1) as an immune-diagnostic biomarker in the pathogenesis of chronic periodontal disease.

OBJECTIVE: Monocyte chemoattractant protein-1 (MCP-1) is an important chemokine responsible for the initiation, regulation and mobilization of monocytes to the active sites of severe periodontal inflammation. The present study aims at evaluating the levels of MCP-1 in GCF, saliva and serum and to analyze the changes following phase I periodontal therapy. Assessment of possible correlations between levels of MCP-1 in the three biological fluids was also done. METHODS: Fifteen healthy and 30 patients of severe chronic periodontitis (diseased) participated in the study. Patients of the diseased group underwent scaling/root planing. Evaluation of PI, GI, PD, CAL and collection of samples of GCF, serum and saliva was done at baseline and 6 weeks following periodontal therapy. MCP-1 levels were quantified in all samples using ELISA. RESULTS: Compared to healthy controls, MCP-1 levels were statistically significantly higher in GCF (p<0.001), saliva (p=0.002) and serum (p<0.001) in subjects with chronic periodontitis. Levels of MCP-1 in all the three fluids decreased significantly in patients after periodontal therapy (p<0.001). There was a significant positive correlation between MCP-1 levels in GCF, saliva and serum in patients of chronic periodontitis both pre (r>0.9) and post-treatment (r>0.6). CONCLUSIONS: The results suggest that levels of MCP-1 in GCF and saliva can be reliable indicators of severity of periodontal destruction and their serum levels reflect the systemic impact of this local inflammatory disease thereby strengthening the reciprocal oro-systemic association.

Plasma homocysteine: an independent or an interactive risk factor for coronary artery disease.

BACKGROUND: Coronary artery disease (CAD) is emerging as a major public health concern in most developing countries. Further conventional risk factors for CAD do not solely account for the increased mortality, particularly in Asians. Recently, increased plasma homocysteine is being considered as a risk factor, but the strength of relationship and interaction of plasma homocysteine with other risk factors is yet obscure. In this study, the association of plasma homocysteine with CAD and other risk factors was estimated. METHODS: In the present study, 100 patients of CAD and 50 controls of both sexes were included. Plasma total homocysteine (tHcy) concentrations were measured using reverse phase high-performance liquid chromatography. RESULTS: Plasma homocysteine concentrations were significantly raised in cases as compared to age-matched controls (16.57+/-6.86 and 11.47+/-5.19 micromol/l, p<0.001). On calculating relative risk (RR) of each factor by univariate analysis smoking, hypertension, plasma cholesterol and homocysteine appeared to be significant risk factors. However, on applying multiple logistic regression only the latter three emerged as independent risk factors (p<0.005). Further, strong interactive effects were observed between homocysteine levels and increasing age, hypertension and smoking. CONCLUSION: An increase in plasma homocysteine concentration confers an independent risk for CAD. It further increases the risk associated with increasing age, smoking and hypertension. Thus, increased homocysteine concentrations are a significant medical problem and effective strategies are urgently required to counter this challenge.