Quantitative analysis of lncRNA in formalin-fixed paraffin-embedded tissues of oral squamous cell carcinoma.

The study evaluated expression profiles of few regulatory lncRNAs in oral squamous cell carcinoma and normal mucosa adjacent to oral cancer using paired fresh frozen and formalin-fixed paraffin-embedded (FFPE) tissues stored at a different duration of time (1-5 years) using real-time quantitative PCR. The quantity and quality of total RNA isolated from FFPE tissues was less compared with that of fresh frozen tissues, which resulted in a noncorrelation of quantification cycle values. Following normalization, the expression of lncRNAs in the paired tissues did not differ significantly. The differential expression of the lncRNAs in the study was consistent with The Cancer Genome Atlas head and neck squamous cell carcinoma database. The study findings demonstrate the possibility of performing accurate quantitative analysis of lncRNAs using short amplicons and standardized real-time quantitative PCR assays in oral squamous cell carcinoma FFPE samples.

Induction of Endometrial HOXA 10 Gene Expression by Vitamin D and its Possible Influence on Reproductive Outcome of PCOS Patients Undergoing Ovulation Induction Procedure.

Background and Objectives: Polycystic ovarian syndrome (PCOS) is one of the most common endocrinopathies in women frequently presenting with anovulatory infertility. Low successful pregnancy and live birth rates even after successful ovulation induction (OI) and in vitro fertilization (IVF) in these patients indicate that endometrial dysfunction may be another important factor contributing to infertility. Vitamin D acting through nuclear receptors induces the expression of various genes required for cell growth and differentiation and plays a crucial role in reproduction. Homeobox 10 (HOXA10) may be one of the potential targets for vitamin D action. HOXA10 gene product promotes the differentiation of endometrial cells, making the endometrium receptive for implantation. The present study was undertaken to determine the effect of circulating vitamin D levels on HOXA10 gene expression in endometrial tissues and its possible influence on the reproductive outcome of PCOS patients undergoing OI procedure. Materials and Methods: A prospective cohort study was conducted on 110 infertile PCOS patients. The patients were divided into two groups: Group 1: Vitamin D ³20 ng/ml, Group 2: Vitamin D <20 ng/ml. Endometrial samples were obtained from 22 patients using pipelle biopsy, used to determine HOXA10 mRNA (messenger ribonucleic acid) expression by quantitative RT-PCR (reverse transcription-polymerase chain reaction) and protein expression by Western blotting. OI was performed using Clomiphene citrate or Letrozole from the 3rd day of the cycle, and patients were followed up for a maximum of five cycles. Attainment of successful pregnancy was considered a positive outcome. Results: Both the groups were similar in mean age and other endocrine parameters. Serum vitamin D levels were significantly low (P < 0.001), and BMI (body mass index) was significantly high (P = 0.032) in group 2 compared to group 1. Endometrial HOXA10 mRNA (by quantitative rtPCR) and protein expression (by western blotting) were significantly low in group 2 compared to group 1. The clinical pregnancy rate was low in group 2 (28.6%) compared to group 1 (42.3%), but this difference was not significant (P = 0.22). On regression analysis adjusted for age and BMI, vitamin D was an independent predictor of successful pregnancy after OI (P = 0.09). Conclusion: Circulating vitamin D levels influence the endometrial HOXA10 gene expression, and this may be reflected on the reproductive outcome of infertile PCOS patients undergoing OI.

INCIDENCE OF INFLAMMATORY BOWEL DISEASE: A SINGLE CENTRE RETROSPECTIVE STUDY.

BACKGROUND: Inflammatory bowel disease (IBD) is a chronic inflammatory disorder affecting the gastrointestinal tract. The etiology of this alarming condition is multifactorial. A Recently increasing trend in IBD is noted in our country. OBJECTIVE: The present study was designed with the main objective to assess the incidence and to identify the associated risk factors including demographic, geographical areas, and dietary patterns of IBD population of Northern of Karnataka viz. Hubli-Dharwad city. METHODS: A retrospective investigation was conducted on a cohort of 226 patients with a working diagnosis of IBD and those who were admitted between 2015 to 2019 the department of gastroenterology, SDMCMS&H. The diagnosis of IBD was made based on clinical, radiological, endoscopic, and histopathologic findings. The patients were categorized into IBD and those who have symptoms suggestive of IBD but did not fit into the diagnostic criteria into, non-IBD groups. The data about of on demography, diet patterns, and laboratory parameters were recorded. RESULTS: Among 226 patients enrolled in this study 2015-2019, IBD was confirmed in 54 Ulcerative colitis - 44 (19.46%), Crohn's disease - 10 (4.42%) patients with varying distribution of disease among different age groups and both genders, Ulcerative colitis (UC) [M: F: 28 (63.6%): 16 (36.4%)] and Crohn's disease (CD) [M: F: 07 (70.0%):03 (30.0%)]. Dietary pattern and other habitats had no significant contribution to illness and its symptoms. Urban (U) and Rural (R) divide was UC [U: R: 32 (72.7%): 12 (27.3%)], CD [U:R:07(70.0%):03(30.0%)] maintained. CONCLUSION: Incidence of IBD was high with UC as compared to CD. The incidence of IBD among patients presenting with symptoms suggestive of IBD is 19.46% with UC being major as compared to CD (4.42%). Male predominant patterns of IBD incidences were noted. Year by year increasing trend in disease burden was observed. The Dietary pattern has no direct correlation with IBD disease prevalence and incidences.

Incidência de doença inflamatória intestinal: estudo retrospectivo de um único centro / Incidence of inflammatory bowel disease: a single centre retrospective study

ABSTRACT Background: Inflammatory bowel disease (IBD) is a chronic inflammatory disorder affecting the gastrointestinal tract. The etiology of this alarming condition is multifactorial. A Recently increasing trend in IBD is noted in our country. Objective: The present study was designed with the main objective to assess the incidence and to identify the associated risk factors including demographic, geographical areas, and dietary patterns of IBD population of Northern of Karnataka viz. Hubli-Dharwad city. Methods: A retrospective investigation was conducted on a cohort of 226 patients with a working diagnosis of IBD and those who were admitted between 2015 to 2019 the department of gastroenterology, SDMCMS&H. The diagnosis of IBD was made based on clinical, radiological, endoscopic, and histopathologic findings. The patients were categorized into IBD and those who have symptoms suggestive of IBD but did not fit into the diagnostic criteria into, non-IBD groups. The data about of on demography, diet patterns, and laboratory parameters were recorded. Results: Among 226 patients enrolled in this study 2015-2019, IBD was confirmed in 54 Ulcerative colitis - 44 (19.46%), Crohn's disease - 10 (4.42%) patients with varying distribution of disease among different age groups and both genders, Ulcerative colitis (UC) [M: F: 28 (63.6%): 16 (36.4%)] and Crohn's disease (CD) [M: F: 07 (70.0%):03 (30.0%)]. Dietary pattern and other habitats had no significant contribution to illness and its symptoms. Urban (U) and Rural (R) divide was UC [U: R: 32 (72.7%): 12 (27.3%)], CD [U:R:07(70.0%):03(30.0%)] maintained. Conclusion Incidence of IBD was high with UC as compared to CD. The incidence of IBD among patients presenting with symptoms suggestive of IBD is 19.46% with UC being major as compared to CD (4.42%). Male predominant patterns of IBD incidences were noted. Year by year increasing trend in disease burden was observed. The Dietary pattern has no direct correlation with IBD disease prevalence and incidences.

Resumo Contexto: A Doença inflamatória intestinal (DII) é uma doença inflamatória crônica que afeta o trato gastrointestinal. A etiologia desta condição alarmante é multifatorial. Uma tendência recentemente crescente na DII é notada em nosso país. Objetivo: O presente estudo foi desenhado com o objetivo principal de avaliar a incidência e identificar os fatores de risco associados, incluindo demográficos, áreas geográficas e padrões alimentares da população com DII do Norte de Karnataka viz. Cidade de Hubli-Dharwad. Metodos: Uma investigação retrospectiva foi realizada em uma coorte de 226 pacientes com diagnóstico de DII e que foram admitidos entre 2015 e 2019 no departamento de gastroenterologia, SDMCMS&H. O diagnóstico de DII foi feito com base em achados clínicos, radiológicos, endoscópicos e histopatológicos. Os pacientes foram categorizados em DII e aqueles que apresentam sintomas sugestivos de DII, mas não se enquadraram nos critérios diagnósticos em grupos sem DII. Os dados sobre a demografia, padrões de dieta e parâmetros laboratoriais foram registrados. Resultados: Entre os 226 pacientes inscritos neste estudo entre 2015-2019, DII foi confirmada em 54 [RCUI - 44 (19,46%), DC - 10 (4,42%)] com distribuição variada da doença entre diferentes faixas etárias e ambos os sexos, colite ulcerativa (RCUI) [M: F: 28 (63,6%):16 (36,4%)] e doença de Crohn (DC) [M: F: 07 (70,0%): 03 (30,0%)]. O padrão alimentar e outros hábitos não tiveram contribuição significativa para a doença e seus sintomas. Urbanos (U) e rurais (R) dividiram-se em RCUI [U: R: 32 (72,7%):12 (27,3%)], DC [07 (70,0%): 03 (30,0%)]. Conclusão A incidência de DII foi elevada para RCUI em relação a DC. A incidência de DII entre os pacientes com sintomas sugestivos de DIB é de 19,46% com a RCUI sendo maior em relação a DC (4,42%). Foram observados padrões predominantes masculinos de incidência de DII. Ano a ano foi observada tendência crescente de carga da doença. O padrão dietético não tem correlação direta com a prevalência e incidências da DII.

An optimized protocol for total RNA isolation from archived formalin-fixed paraffin-embedded tissues to identify the long non-coding RNA in oral squamous cell carcinomas.

Approximately 93% of the human genome is translated into RNAs, of which only 2% code for proteins and the rest 98% are noncoding RNAs. Long non-coding RNAs (lncRNAs) are a class of non-coding RNAs of > 200 nucleotides length that are emerging as novel players in the field of cancer diagnostics or prognostics. Recently, lncRNAs are known to be associated with oral squamous cell carcinomas (OSCC). The demonstration of stable lncRNA has been a challenge in formalin-fixed paraffin-embedded tissues (FFPE). The survivability and expression level of lncRNA in FFPE tissues compared with fresh tissues is not well documented in the literature. Hence, we designed the current pilot study with the main aim to optimize modified TRI (Total RNA isolation) reagent RNA isolation protocol to identify the lncRNA expression in archived FFPE tissues of OSCC in comparison to the standard RNA isolation kit method. The findings of our study demonstrated that the RNA quantity and quality were comparatively better with the optimized TRI reagent modified protocol than the standard RNA isolation kit method. Furthermore, ct (cycle threshold) values after reverse-transcription and qRT-PCR (Quantitative Real time PCR) were comparable and almost equal in both the methods for normal mucosa (control) and OSCC samples.