Effect of Non-Coding RNA on Post-Transcriptional Gene Silencing of Alzheimer Disease

A large amount of hidden biological information is contained in the human genome, which is not expressed or revealed in the form of proteins; the usual end product form of gene expression. Instead, most of such information is in the form of non-coding RNAs (ncRNAs). ncRNAs correspond to genes that are transcribed, but do not get translated into proteins. This part of the genome was, till recently, considered as ‘junk’. The term ‘junk’ implied lack of any discernible function of these RNA. More than 98% of the human genomic size encompasses these non-coding RNAs. But, recent research has evidently brought out the indispensible contribution of non-coding RNA in controlling and regulating gene expression. ncRNA such as siRNAs and microRNAs have been reported to greatly help in causing post-transcriptional gene silencing (PTGS) in cells through RNA interference (RNAi) pathway. In this work, we have investigated the possibility of using siRNAs and microRNAs to aid in gene silencing of early onset Alzheimer’s disease genes. 
Alzheimer’s disease specific mutations and their corresponding positions in mRNA have been identified for six genes; Presenilin-1, Presenilin-2, APP (amyloid beta precursor protein), APBB3, BACE-1 and PSENEN. 

Small interfering RNAs (siRNAs) that can cause PTGS through RNA interference pathway have been designed. RNA analysis has been done to verify complementarity of antisense siRNA sequence with target mRNA sequence. Interaction studies have been done computationally between these antisense siRNA strands and seven Argonaute proteins. From the interaction studies, only one of the seven Argonaute proteins; 1Q8K, was found to have interaction with the siRNAs indicating the importance and uniqueness of this particular protein in RISC (RNA induced silencing complex). 

The interaction studies have been carried out for the microRNAs also. Out of the 700 mature human microRNAs collected, 394 microRNAs have been identified to show partial complementarity with their target sequence on PSEN-1 mRNA. Of these 394, five microRNAs have shown partial complementarity to early onset Alzheimer’s disease specific mutations in PSEN-1 mRNA. Interaction studies have been done between these microRNAs and Argonaute proteins. Thus, design, characterization and analysis of ncRNAs that contribute to post transcriptional gene silencing of Alzheimer’s disease have been achieved.
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Cervical Cancer screening program in Hyderabad and surrounding peri-urban areas, South India: prevalence of high risk HPV subtypes

AbstractPurposeCervical screening programs were conducted in urban and peri-urban areas of Telangana and Andhra Pradesh, during the period January 2012–March 2014. Our work was aimed at (I) identifying women with abnormal cervix upon visual inspection and pap cytology testing (II) determining the presence of HPV infection in the individual, along with sub-typing in severe pathologies and cancer, which is an established risk factor for cervical malignancy reported world-over. Methods      A total of 530 eligible women (based inclusion criteria for participation) were screened by gynecologist. Pap smears were collected on slides and cervical swab cells were collected in a saline solution for subsequent cytological and molecular analysis. The study was approved by our Ethics committee and informed consent was obtained from the subjects. Personal and medical history of each of the eligible participants was recorded in a well-designed proforma. Data tabulation and statistical analysis was performed using MS Excel and MedCalc software.ResultsOur program witnessed an incremental increase in the number of women accessing screening after counselling for awareness of the importance of testing. A total of 530 women were screened for Pap smear cytology. The mean age of women in the study population was 38.73yrs (+10.82). 16.41% cases showed a normal cytology, 43.39% had an inflammatory smear, 26.22% showed infection, 4.15% showed reactive cellular changes, 6.41% showed atrophy, 0.75% showed ASCUS, 1.88% showed LSIL, 0.37% showed HSIL, 0.37% showed squamous cell carcinoma. Prevalence of HPV in our study population was 14.7% (n=530). 1.8% of our subjects showed the presence of high-risk HPV subtypes; all of them were associated with an abnormal Pap cytology. HPV was shown to be associated with an infectious pap, RCC, ASCUS, HSIL, SCC (p0.05) when compared to normal cytology (p=0.05). ConclusionsAwareness and the importance of cervical examination is low. Health camps need to focus on counselling subjects about its benefits to improve their participation and ensure success of screening programs. The significant association of HPV infection with abnormal pathologies (p=0.05) and the presence of hr-HPV subtypes other than 16 and 18 (that are used for vaccination) draws attention to the need to evaluate the subtypes prevalent in our population and apply this information to cervical vaccination schemes.Keywords Cervical, Pap cytology, HPV, High-risk subtypes Abbreviations HPV- Human Papilloma Virus, hr-HPV-high risk Human Papilloma Virus, RCC-Reactive Cellular Changes, ASCUS- Atypical Squamous Cells of Undetermined Significance, LSIL-Low Grade Squamous Intraepithelial Lesion, HSIL-High Grade Squamous Intraepithelial Lesion, CIN-Cervical intraepithelial neoplasia, SCC-Squamous cell carcinoma

(CTG)_n Expansion at DMPK locus seen only in muscle tissue: A novel case

937-940Triplet repeat expansion in 3 untranslated region of myotonic dystrophy protein kinase (DMPK) gene has been implicated as causative in myotonic dystrophy (DM). In cases of DM, high levels of somatic in stability have been reported, in which inter-tissue repeat length differences as large as 3000 repeats have been observed. This study highlights the inter-tissue (CTG)n expansion variability at the DMPK locus. Molecular analysis of DMPK gene, encompassing the triplet repeat expansion, was carried out in 31 individuals (11 clinically identified DM patients, 20 controls). All controls showed a 2.1 kb band (upto 35 CTG repeats), while four cases exhibited an expansion (>50 repeats ). A novel observation was made in one case, wherein the D A from lymphocytes showed a normal 2.1 kb band while the muscle tissue DNA from the same patient was heterozygous for normal and 4.3 kb band (> 700 repeats). Our results suggested that because inter-tissue variability existed in the (CTG)n repeat number at DMPK locus, an attempt should be made to evaluate affected tissue along with blood wherever possible prior to making a final diagnosis. This is important not only for diagnosis and prenatal analysis, but also while providing genetic counseling to families

Cardiovascular diseases: Interplay of epigenetics

Several association studies have been carried out to elucidate the role of genetic variants in cardiovascular diseases (CVDs), while studies on the epigenome regulating gene expression changes are helping to understand the development of disease and factors promoting such changes. This review summarizes the different epigenetic aspects involved in cardiac development and disease along with current therapeutic interventions

Analysis of Single Nucleotide Polymorphisms on Locus 13q33.1-34 in Multigenerational Families of Cleft Lip Palate using MassArray

BACKGROUND: Cleft lip palate is a common congenital anomaly with multifactorial etiology. Many high-risk markers at different loci were reported to be involved in its etiology. Advanced genetic research led to the discovery of evidence of a new linkage on 13q33.1-34 region at marker rs1830756 in two multigenerational Indian families. However, no further study was reported to confirm or validate this linkage in other families. Hence, the present study was designed.METHODS: Twenty multigenerational families affected by non-syndromic cleft lip palate were selected for the study. Polymorphisms, rs1830756, rs1323672, rs1935135 of FAM155A gene; rs1961495, rs953386, rs1411040 of COL4A1 gene; and rs726449, rs984300 of MYO16 gene were selected. Genomic DNA was isolated and sent for genetic analysis by single nucleotide polymorphism (SNP) genotyping using the MassArray method. Statistical analysis of the genomic data was done by PLINK. Bonferroni correction was applied and haplotype analysis was done using Haploview software.RESULTS: Polymorphisms followed the Hardy Weinberg Equilibrium. In the allelic association, all the polymorphisms analysed showed no statistical significance. Hence, there was no significant difference in the allelic frequencies between non-syndromic cleft lip palate patients and healthy controls. The odds ratio was not more than 1.6 for all the SNPs. Haplotype analysis showed that haplotypes were not significantly higher in non-syndromic cleft patients than in control subjects.CONCLUSION: There is no association between SNPs analysed in the locus 13q33.1-34 with cleft lip palate.KEYWORDS: cleft lip palate, chromosome, polymorphis