Dysphagia Continues to Impact Recovery at One Year After Stroke-An Observational Study.

OBJECTIVES: Dysphagia is one of the most common medical complication after acute stroke, which can impact hospital stay and discharge outcome. Here we sought to study the predictors and 1 year outcome of patients with acute post stroke dysphagia. METHODS: Single centre hospital based observational study of all acute ischemic stroke patients who had undergone formal swallow assessment within 24 hours of admission with a 1 year completed follow-up were recruited by screening of medical records. Clinical, imaging and swallow assessment details were extracted as per proforma. 3 month and 1 year outcome were assessed using modified Rankin scale. Correlations were made with clinical and imaging findings, in hospital worsening and dysphagia at discharge with short and long term functional outcome. RESULTS: We had 469 patients included in our study, with a mean age 61. 04(±19. 09) years and median NIHSS 9. 52(IQR 4). 56. 75% of AIS patients had some degree of dysphagia at admission. We found that admission stroke severity and in-hospital worsening were independently predictive of severe swallow dysfunction at discharge. At 3-4 weeks after stroke, only 20.27% of the patients with moderate-severe dysphagia at baseline has persistent swallow deficits requiring modification of feeds. Dysphagia continued to have a significant association with outcome at 1 year, independent of admission stroke severity. CONCLUSIONS: Majority of patients with acute post stroke dysphagia recover well within 3-4 weeks after stroke. Patients with post stroke dysphagia had more in hospital neurological worsening. Post stroke dysphagia continued to have an impact on functional outcome up to 1 year after stroke.

STIL Endows Oncogenic and Stem-Like Attributes to Colorectal Cancer Plausibly by Shh and Wnt Signaling.

The discovery of a potent gene regulating tumorigenesis and drug resistance is of high clinical importance. STIL is an oncogene; however, its molecular associations and role in colorectal oncogenesis are unknown. In this study, we have explored the role of STIL gene in tumorigenesis and studied its molecular targets in colorectal cancer (CRC). STIL silencing reduced proliferation and tumor growth in CRC. Further, STIL was found to regulate stemness markers CD133 and CD44 and drug resistant markers thymidylate synthase, ABCB1, and ABCG2 both in in-vitro and in-vivo CRC models. In addition, high expression of STIL mRNA was found to be associated with reduced disease-free survival in CRC cases. Interestingly, we observed that STIL-mediated regulation of stemness and drug resistant genes is not exclusively governed by Sonic hedgehog (Shh) signaling. Remarkably, we found STIL regulate ß-catenin levels through p-AKT, independent of Shh pathway. This partially answers Shh independent regulatory mechanism of cancer stem cell (CSC) markers by STIL. Our study suggests an instrumental role of STIL in molecular manifestation of CRC and progression.

Low Bifidobacterium Abundance in the Lower Gut Microbiota Is Associated With Helicobacter pylori-Related Gastric Ulcer and Gastric Cancer.

Helicobacter pylori infection in stomach leads to gastric cancer, gastric ulcer, and duodenal ulcer. More than 1 million people die each year due to these diseases, but why most H. pylori-infected individuals remain asymptomatic while a certain proportion develops such severe gastric diseases remained an enigma. Several studies indicated that gastric and intestinal microbiota may play a critical role in the development of the H. pylori-associated diseases. However, no specific microbe in the gastric or intestinal microbiota has been clearly linked to H. pylori infection and related gastric diseases. Here, we studied H. pylori infection, its virulence genes, the intestinal microbiota, and the clinical status of Trivandrum residents (N = 375) in southwestern India by standard H. pylori culture, PCR genotype, Sanger sequencing, and microbiome analyses using Illumina Miseq and Nanopore GridION. Our analyses revealed that gastric colonization by virulent H. pylori strains (vacAs1i1m1cagA+) is necessary but not sufficient for developing these diseases. Conversely, distinct microbial pools exist in the lower gut of the H. pylori-infected vs. H. pylori-non-infected individuals. Bifidobacterium (belonging to the phylum Actinobacteria) and Bacteroides (belonging to the phylum Bacteroidetes) were present in lower relative abundance for the H. pylori+ group than the H. pylori- group (p < 0.05). On the contrary, for the H. pylori+ group, genus Dialister (bacteria belonging to the phylum Firmicutes) and genus Prevotella (bacteria belonging to the phylum Bacteroidetes) were present in higher abundance compared to the H. pylori- group (p < 0.05). Notably, those who carried H. pylori in the stomach and had developed aggressive gastric diseases also had extremely low relative abundance (p < 0.05) of several Bifidobacterium species (e.g., B. adolescentis, B. longum) in the lower gut suggesting a protective role of Bifidobacterium. Our results show the link between lower gastrointestinal microbes and upper gastrointestinal diseases. Moreover, the results are important for developing effective probiotic and early prognosis of severe gastric diseases.

A Study of the Developing Paediatric Skullbase Anatomy and its Application to Endoscopic Endonasal Approaches in Children.

OBJECTIVE: The sinonasal anatomy in growing children undergoes change in size along with progressive pneumatization, this is of particular significance in endoscopic endonasal surgery. We aim to measure and quantify the sinonasal dimensions in the Indian paediatric population, which are relevant to skull base surgeons. MATERIAL AND METHODS: This is a retrospective radio anatomical study of sinonasal measurements and volumetric analysis of the sphenoid sinus performed on archived CT images of children less than 18 years of age. RESULTS: CT scan images of 110 patients (male, 68; female, 42) were included for the study. The number of patients in each age groups was as follows [0-6 years, 17; 7-9 years, 20; 10-12 years, 27; 13-15 years, 29; 16-18 years, 17]. The distance from the nares to the sphenoid and from the sphenoid to the sella was significantly greater in children of 13-15 years (69.4 ± 5.2 mm) as compared to children less than six years (62.6 ± 6.7 mm) (P < 0.003). The volume of the sphenoid in children between 0 and 6 years was 4641.4 ± 1924.7 mm3. The pneumatized sphenoid volume in the same age group was 1655 ± 1631.1 mm3. In older children between 13 and 15 years, the total volume of sphenoid sinus was 11732.8 ± 2614.4 mm3. The volume of pneumatization in the sphenoid sinus in this group was 6287.5 ± 2157.9 mm3. The total volume of the sphenoid sinus (Pearson coefficient (r) = 0.704, P < 0.001) and the volume of pneumatization of the sphenoid sinus was also seen to have a positive correlation to the age of the child (r = 0.62, P < 0.0001). The narrowest distance between both the internal carotid arteries was seen at the level of the proximal dural ring. In children less than six years of age it was 13.4 ± 2.0 mm, a significant change was seen by the age of 10-12 years where this distance was 15.6 ± 2.2 mm (P = 0.036). CONCLUSION: The sinonasal anatomy shows progressive development between the age of 6 to 15 years after which it plateaus. The pneumatization in young children may be incomplete, which necessitates drilling in the sphenoid sinus. The intercarotid distance was not seen to be a hindrance for endoscopic endonasal surgery. None of the measurements of the skull-base, made in this study appear to restrict endoscopic endonasal surgery in children. However, a meticulous preoperative assessment of the CT scan may be needed for optimal surgical outcome.

Using open-source data to explore distribution of built environment characteristics across Kerala, India.

BACKGROUND: Built environment characteristics in the neighborhood are of utmost priority for a healthy lifestyle in the fast-urbanizing countries. These characteristics are closely linked to the disease burden and challenges in low- and middle-income countries (LMICs), which have been unexplored using open-source data. The present technology offers online resources and open source software that enable researchers to explore built environment characteristics with health and allied phenomena. OBJECTIVES: This article intends to delineate methods to capture available and accessible objective built environment variables for a state in India and determine their distribution across the state. METHODS: Built environment variables such as population density and residential density were collated from the Census of India. Safety from crime and traffic were captured as crime rates and pedestrian accident rates, respectively, acquired from State Crime Records Bureau. Greenness, built-up density, and land slope were gathered from open-source satellite imagery repository. Road intersection density was derived from OpenStreetMap. Processing and analysis differed for each dataset depending on its source and nature. RESULTS: Each variable showed a distinct pattern across the state. Population and residential density were found to be closely related to each other across both districts and subdistricts. They were both positively related to crime rates, pedestrian accident rates, built-up density, and intersection density, whereas negatively related to land slope and greenness across the subdistricts. CONCLUSION: Delineating the distribution of built environment variables using available and open-source data in resource-poor settings is a first in public health research among LMICs. Cost-effectiveness and reproducible nature of open-source solutions could equip researchers in resource-poor settings to identify built environment characteristics and patterns across regions.

Spatial Clusters of Diabetes and Physical Inactivity: Do Neighborhood Characteristics in High and Low Clusters Differ?

This study aims to find spatial clusters of diabetes and physical inactivity among a sample population in Kerala, India, and evaluate built environment characteristics within the high and low spatial clusters. Spatial clusters with a higher and lower likelihood of diabetes and physical inactivity were identified using spatial scan statistic at various radii. Built environment characteristics were captured at panchayat level and 1600 m buffer around participant location using Geographical Information Systems. Comparison of sociodemographic and built environment factors was carried out for participants within high and low spatial clusters using t tests. Ten high and 8 low spatial clusters of diabetes and 17 high and 23 low spatial clusters of physical inactivity were identified in urban and rural areas of Kerala. Significant differences in built environment characteristics were consistent for low spatial clusters of diabetes and physical inactivity in the urban scenario. Built environment characteristics were found to be relevant in both urban and rural areas of Kerala. There is an urgent call to explore spatial clustering of non-communicable diseases in Kerala and undo the one-size-fits-all approach for prevention and control of non-communicable diseases.

Lack of association of mirSNP rs11174811 in AVPR1A gene with arterial blood pressure and hypertension in South Indian population.

Epigenetic regulation of arterial blood pressure mediated through mirSNPs in renin-angiotensin aldosterone system (RAAS) genes is a less explored hypothesis. Recently, the mirSNP rs11174811 in the 3'UTR of the AVPR1A gene was associated with higher arterial blood pressure in a large study population from the Study of Myocardial Infarctions Leiden (SMILE). The aim of the present study was to replicate the association of mirSNP rs11174811 with blood pressure outcomes and hypertension in a south Indian population. Four hundred and fifteen hypertensive cases and 416 normotensive controls were genotyped using a 5' nuclease allelic discrimination assay. Logistic regression was used to test the association of mirSNP rs11174811 with the hypertension phenotype. Censored normal regression was used to test the association of the polymorphism with continuous blood pressure outcomes such as systolic and diastolic blood pressure. The mirSNP rs11174811 did not show any significant association with hypertension. The adjusted odds ratio was 1.02, with 95% CI of 0.72 to 1.45 (p = 0.909). The mean systolic and diastolic blood pressure values were not significantly different across the three genotypic groups, between hypertensives and normotensives, or when stratified by gender. Despite having a similar minor allele frequency (MAF) of 14.5% compared with the SMILE cohort, our results did not support an association of the mirSNP rs11174811 with the hypertension phenotype or with continuous blood pressure outcomes in the south Indian population.

Folate, vitamin B12, homocysteine and polymorphisms in folate metabolizing genes in children with congenital heart disease and their mothers.

BACKGROUND/OBJECTIVES: The objective of the study was to assess the role of variations in serum folate, vitamin B12, homocysteine and the presence of genetic polymorphisms as risk factors for congenital heart disease (CHD) in children. SUBJECTS/METHODS: A total of 32 children with CHD, and their mothers and 32 normal children and their mothers formed the study and control groups, respectively. Serum folate, vitamin B12 and homocysteine as well as genetic polymorphisms MTHFR C677→︀T, MTHFR A1298→︀C, MTR A2756→︀G and MTRR A66→︀G were assessed. RESULTS: Low serum folate and genetic polymorphisms MTHFR C677→︀T and MTRR A66→︀G among children and their mothers and high homocysteine among mothers were noted as risk factors for CHD (P<0.05). Vitamin B12 levels were normal and showed no association. Presence of MTHFR C677→︀T and MTRR A66→︀G, both concurrently among children as well as mothers and simultaneously among mother-child pairs, showed several fold increase in the risk for CHD. On multivariate analysis, the risk factors noted for CHD were presence of MTHFR C677→︀T among children and their mothers and MTRR A66→︀G among mothers. Analyses for nutrient-gene interaction revealed significant associations between low serum folate and high serum homocysteine levels, and the presence of selected genetic polymorphisms. CONCLUSIONS: Low serum folate, high homocysteine and presence of selected genetic polymorphisms among children and their mothers were noted as risk factors for CHD. Nutrient-gene interaction being a modifiable risk factor, the study recommends the use of peri-conceptional folate supplementation with vitamin B12 sufficiency for primary prevention of CHD.

Evaluating genomic DNA extraction methods from human whole blood using endpoint and real-time PCR assays.

The extraction of genomic DNA is the crucial first step in large-scale epidemiological studies. Though there are many popular DNA isolation methods from human whole blood, only a few reports have compared their efficiencies using both end-point and real-time PCR assays. Genomic DNA was extracted from coronary artery disease patients using solution-based conventional protocols such as the phenol-chloroform/proteinase-K method and a non-phenolic non-enzymatic Rapid-Method, which were evaluated and compared vis-a-vis a commercially available silica column-based Blood DNA isolation kit. The appropriate method for efficiently extracting relatively pure DNA was assessed based on the total DNA yield, concentration, purity ratios (A260/A280 and A260/A230), spectral profile and agarose gel electrophoresis analysis. The quality of the isolated DNA was further analysed for PCR inhibition using a murine specific ATP1A3 qPCR assay and mtDNA/Y-chromosome ratio determination assay. The suitability of the extracted DNA for downstream applications such as end-point SNP genotyping, was tested using PCR-RFLP analysis of the AGTR1-1166A>C variant, a mirSNP having pharmacogenetic relevance in cardiovascular diseases. Compared to the traditional phenol-chloroform/proteinase-K method, our results indicated the Rapid-Method to be a more suitable protocol for genomic DNA extraction from human whole blood in terms of DNA quantity, quality, safety, processing time and cost. The Rapid-Method, which is based on a simple salting-out procedure, is not only safe and cost-effective, but also has the added advantage of being scaled up to process variable sample volumes, thus enabling it to be applied in large-scale epidemiological studies.

Oncogenic microRNAs as biomarkers of oral tumorigenesis and minimal residual disease.

OBJECTIVES: Classical diagnostic methods are not sensitive enough in detecting oral lesions that may progress to cancer and in assessing minimal residual disease (MRD) in oral surgical margins. Altered expression of microRNAs (miRNAs) contributes to human cancer, including oral cancer. Although there are many studies on microRNAs in oral cancer, there is no reported study comparing the expression of microRNAs during oral tumor progression and in oral surgical margins. MATERIALS AND METHODS: This study analyzed the expression of 72 miRNAs that were reported (till June 2011) to be differentially expressed in oral cancer, during phases of oral cancer progression and in oral surgical margins. RESULTS: Of the 72 miRNAs analyzed, four (hsa-miR-125a, hsa-miR-184, hsa-miR16 and hsa-miR-96) showed a common pattern of expression in both sets of tissues. We further analyzed the downstream target genes of hsa-miR-16 BCL2 and CCND1. The in silico network analysis of these four microRNAs and their target genes revealed presence of genes involved in tumor progression and transcription factors. CONCLUSIONS: The findings suggest that the combinatorial regulation by these miRNAs and their target transcription factors might play a substantial role in oral tumorigenesis. Here we report for the first time that a decreased expression of hsa-miR-125a, hsa-miR-184 and hsa-miR-16 and an increased expression of hsa-miR-96 could be useful in predicting oral tumorigenesis and importantly in the detection of MRD and decision-making process for postoperative treatment modalities.

C-T variant in a miRNA target site of BCL2 is associated with increased risk of human papilloma virus related cervical cancer--an in silico approach.

MicroRNAs control gene expression at the posttranscriptional level by base-pairing to the 3'-UTR of their target mRNAs, thus leading to mRNA degradation of protein fabrication. We hypothesize, SNPs within miRNAs and their targets could be of significance to an individual's risk of developing cancer. We analyzed in silico SNP information on cervical cancer associated aberrant alleles and further investigated this in a case-control study by examining eleven SNPs from different genes. It was observed that a C to T polymorphism in putative miRNA target site of BCL2 was significantly conspicuous for the aberrant SNP allele in cancer tissues as compared to controls. This study provides evidence that SNPs in miRNA-binding sites may play an important role in increasing risk of cancer. The results also paves way for future studies to validate these results in other well-characterized populations as well as to explore the biological significance of these particular SNPs.