Canonical Wnt pathway gene expression and their clinical correlation in oral squamous cell carcinoma.

AIM: The aim of this study is to explore the prognostic significance and clinicopathological correlations of the Wnt pathway genes in a cohort of surgically treated patients with oral squamous cell carcinoma (OSCC) patients. SETTINGS AND DESIGN: A prospective genetic study on patients with OSCC was carried out during the period from July 2014 to January 2016. Informed consent from patients and institutional ethical approval for the study was obtained and the guidelines were strictly followed for collection of samples. SUBJECTS AND METHODS: Clinical data and mRNA expression analysis of ten genes in the canonical Wnt pathway were evaluated and their relationships with clinical and demographic variables were studied in 58 tissue samples. Wnt-3a, ß-catenin, secreted frizzled-related proteins sFRP-1, sFRP-2, sFRP-4, sFRP-5, Wnt inhibitory factor 1, dickkopf-1, c-MYC, and cyclin-D1 from cancer (n = 29) and normal (n = 29) tissue samples were investigated using quantitative reverse transcription-polymerase chain reaction. STATISTICAL ANALYSIS: Descriptive statistics were used to summarize the sample characteristics and clinical variables. If the data were normal, then parametric tests were used; otherwise, nonparametric alternatives were used. All the analyses were carried out using SPSS version 23.0 (IBM SPSS Inc., USA). RESULTS: Expression of sFRP-1, sFRP-2, and sFRP-5 in control samples and expression of c-MYC and cyclin D1 in cancer samples showed statistical significance. Significant expression of Wnt3A was observed among patients who had recurrence and were deceased. CONCLUSION: Wnt3A, ß-catenin, and cyclin D1 are recognized as key components of Wnt/ß-catenin signaling. However, in this study, there was no significant expression of all the three genes in OSCC. The proto-oncogene c-MYC showed statistically significant upregulation in cancer tissue samples suggesting that the OSCC among South Indian population is primarily not mediated by the canonical Wnt signaling pathway.

Association between specific periodontal pathogens, Toll-like receptor-4, and nuclear factor-κB expression in placental tissues of pre-eclamptic women with periodontitis.

AIM: The aim of the present study was to determine the association between the presence of specific periodontal pathogens, Toll-like receptor-4 (TLR-4), and nuclear factor-κB (NF-κB) expression in the placental tissues of pre-eclamptic women. METHODS: Antenatal periodontal screening was performed in 25 normotensive pregnant women and 25 pre-eclamptic women. Subgingival plaque and placental tissue samples were collected from both groups and screened for the presence of Porphyromonas gingivalis (P. gingivalis), Tannerella forsythia, Aggregatibacter actinomycetemcomitans, and Prevotella intermedia (P. intermedia) using real-time polymerase chain reaction. The placental samples were also analyzed to quantify TLR-4 and NF-κB expression. RESULTS: The subgingival plaque samples of pre-eclamptic women showed significantly higher frequencies of P. intermedia. In the placental samples, P. gingivalis, P. intermedia, and the expression of TLR-4 and NF-κB were found to be at significantly higher levels compared to normotensive pregnant women. Using linear regression analysis, the expression of TLR-4 was significantly influenced by the presence of P. gingivalis (coefficient=3.176, 95% confidence interval [CI]: 367-5.986) and P. intermedia (coefficient=2.886, 95% CI: 0.77-5.696), whereas NF-κB expression was influenced only by the presence of P. intermedia (coefficient=2.220, 95% CI: 0.051-4.388) in the placental tissues of pre-eclamptic women. CONCLUSION: An association exists between P. gingivalis and P. intermedia with increased TLR-4 and NF-κB expression in the placenta of pre-eclamptic women with periodontitis.

Evidence Linking the Role of Placental Expressions of Peroxisome Proliferator-Activated Receptor-γ and Nuclear Factor-Kappa B in the Pathogenesis of Preeclampsia Associated With Periodontitis.

BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)-γ activation leads to suppression of production of a broad range of proinflammatory molecules. It plays a role in differentiation of trophoblasts and helps in normal placentation and formation of vascular exchange interface. Activation of nuclear factor-kappa (NF-κ) B triggers proinflammatory molecules inducing abnormal placentation and premature labor. This study aims to explore expression of PPAR-γ and NF-κB in placentas of women with periodontitis-associated preeclampsia compared with that in normotensive pregnant women. METHODS: Fifty pregnant women were included. Twenty-five were controls (normotensive pregnant women) and 25 were pregnant women with preeclampsia, including those with gestational hypertension. Demographic data, pregnancy characteristics, and periodontal parameters were recorded, including: 1) plaque index; 2) gingival index; 3) bleeding on probing (BOP); 4) probing depth; and 5) attachment loss (AL). Placental tissue samples were collected from both groups and analyzed to quantify expression of PPAR-γ and NF-κB using real-time polymerase chain reaction. RESULTS: BOP and AL were significantly higher in pregnant women with preeclampsia compared with normotensive pregnant women (P <0.05). Expression of PPAR-γ was downregulated in patients with preeclampsia compared with that of healthy normotensive patients, which was statistically significant (P <0.05), whereas NF-κB was significantly activated (P <0.05) in pregnant women with preeclampsia compared with normotensive pregnant women. CONCLUSIONS: Higher periodontal disease prevalence is found among pregnant women with preeclampsia, with increased percentage of sites with BOP and greater AL. This study provides novel information on host response to systemic inflammation induced by periodontal pathogens through mechanisms involving downregulation of PPAR-γ and increased activation of NF-κB.

Myocilin mutations among POAG patients from two populations of Tamil Nadu, South India, a comparative analysis.

PURPOSE: Primary open angle glaucoma (POAG) is the most common type of glaucoma. Among the POAG genes identified so far, myocilin (MYOC) is the most frequently mutated gene in POAG patients worldwide. The MYOC Gln48His mutation is unique among Indian POAG patients. This mutation has not been observed in some populations within India and in other populations worldwide. The objectives of this work were to characterize and compare the mutation spectrum among POAG patients from two places of South India and identify the occurrence and prevalence of Gln48His mutation in our study populations. METHODS: One hundred-one (101) POAG patients from Chennai, South India were recruited for the study. Earlier, 100 patients from the southernmost part of India, Kanyakumari district, were screened. MYOC was screened by polymerase chain reaction based single stand conformation polymorphism (PCR-SSCP) methodology. DNA sequencing of deviant samples was performed. Secondary structures of the proteins with amino acid sequence variations were predicted. RESULTS: The mutation frequency of MYOC among POAG patients in Chennai was 2%. Three types of mutations were observed. The MYOC Gln48His mutation was observed among 2 POAG patients from Chennai. However, absence of this mutation among patients from Kanyakumari suggests possible involvement of demographic factors in disease causation via this mutation. Two heterozygous sequence variants, Thr353Ile and Asn480Lys, in the same exon (exon III) of MYOC were observed in one POAG patient who had a severe disease phenotype. This is the first such report of a compound heterozygote individual with two mutations in the same exon of MYOC. CONCLUSIONS: The presence of mutations at a rate similar to other studies suggests the causative role of MYOC among POAG patients from Chennai. Screening of more patients and families from all parts of India is required to identify the actual frequency of the Gln48His mutation and thus highlight its importance. The compound heterozygote with a severe disease phenotype reiterates the importance of MYOC in certain POAG patients.