Mutational Analysis of the Poly(ADP-Ribosyl)ation Sites of the Transcription Factor CTCF Provides an Insight into the Mechanism of Its Regulation by Poly(ADP-Ribosyl)ation

ABSTRACT Poly(ADP-ribosyl)ation of the conserved multifunctional transcription factor CTCF was previously identified as important to maintain CTCF insulator and chromatin barrier functions. However, the molecular mechanism of this regulation and also the necessity of this modification for other CTCF functions remain unknown. In this study, we identified potential sites of poly(ADP-ribosyl)ation within the N-terminal domain of CTCF and generated a mutant deficient in poly(ADP-ribosyl)ation. Using this CTCF mutant, we demonstrated the requirement of poly(ADP-ribosyl)ation for optimal CTCF function in transcriptional activation of the p 19ARF promoter and inhibition of cell proliferation. By using a newly generated isogenic insulator reporter cell line, the CTCF insulator function at the mouse Igf2-H19 imprinting control region (ICR) was found to be compromised by the CTCF mutation. The association and simultaneous presence of PARP-1 and CTCF at the ICR, confirmed by single and serial chromatin immunoprecipitation assays, were found to be independent of CTCF poly(ADP-ribosyl)ation. These results suggest a model of CTCF regulation by poly(ADP-ribosyl)ation whereby CTCF and PARP-1 form functional complexes at sites along the DNA, producing a dynamic reversible modification of CTCF. By using bioinformatics tools, numerous sites of CTCF and PARP-1 colocalization were demonstrated, suggesting that such regulation of CTCF may take place at the genome level. </jats:p

Tissue localization of collagenase and leucine aminopeptidase in the bovine filarial parasite Setaria cervi

BACKGROUND: Like other helminth proteases, filarial proteases have also been shown to require for parasite survival inside the host and mediate various physiologic processes such as tissue invasion, feeding, embryogenesis and host immune evasion. Many of these proteases have shown potential for vaccines and chemotherapeutic agents against active filarial infections. Setaria cervi is a bovine filarial parasite and serves as a good parasite model for the studies in lymphatic filariasis. Recently, a 175 kDa collagenase and leucine aminopeptidase (LAP) have been purified and characterized from the bovine filarial parasite S. cervi and shown to be potential vaccine candidate and diagnostic marker, respectively for human lymphatic filariasis. However, their tissue localizations and putative roles in the parasite biology have not yet been examined and thus remain unclear. Therefore, the current study attempts to localize and explore the putative roles of these two enzymes in S. cervi. METHODS: The tissue distributions of 175 kDa collagenase and leucine aminopeptidase in S. cervi were examined by immunohistochemical and histochemical methods, respectively. Immune sera obtained from the jirds immunized with collagenase served as primary antibody, rabbit anti-mouse IgG-HRP conjugate as secondary antibody and DAB as the substrate for the immunostaining of collagenase. Leu-βNA was used as the substrate for the histochemical staining of LAP. RESULTS: Both the collagenase and LAP were present in the body wall; however, they differ in their distribution pattern in different layers of body wall. Collagenase was mainly localized in epicuticle, cuticle, syncytial hypodermis and the nerve cord region whereas LAP was more concentrated in epicuticle, longitudinal muscle layers and almost absent or very faintly stained in syncytial hypodermis and nerve cord region. Both collagenase and LAP showed their common distributions in intestine, uterus and mature eggs, growing embryos and mf. Very strong immunostaining of collagenase in the outer body surface of the parasite indicates its major role in host-parasite relationship whereas the presence of LAP in muscular region suggests its role in tissue remodeling. The common presences of collagenase and LAP in the S. cervi intestine, ovary, uterus, eggs and mf suggest that they also have collaborative roles in molting, nutrition and embryogenesis. The data obtained on their immunological characterizations and their presence in important parasite organs give strong indication that they are critical for the survival of filarial parasite and thus can be good vaccine candidates and/or diagnostic markers for human lymphatic filariasis. CONCLUSION: The manuscript reports for the first time the tissue distribution of collagenase and LAP in the bovine filarial parasite S. cervi and discuss their putative roles in vivo. Our findings also open the avenue to examine the roles of these two proteases in vivo, which will require further experiments like using their natural substrates and/or specific inhibitors in each tissues

Somatically acquired hypomethylation of IGF2 in breast and colorectal cancer

The imprinted insulin-like growth factor 2 (IGF2) gene is expressed predominantly from the paternal allele. Loss of imprinting (LOI) associated with hypomethylation at the promoter proximal sequence (DMR0) of the IGF2 gene was proposed as a predisposing constitutive risk biomarker for colorectal cancer. We used pyrosequencing to assess whether IGF2 DMR0 methylation is either present constitutively prior to cancer or whether it is acquired tissue-specifically after the onset of cancer. DNA samples from tumour tissues and matched non-tumour tissues from 22 breast and 42 colorectal cancer patients as well as peripheral blood samples obtained from colorectal cancer patients [SEARCH (n=case 192, controls 96)], breast cancer patients [ABC (n=case 364, controls 96)] and the European Prospective Investigation of Cancer [EPIC-Norfolk (n=breast 228, colorectal 225, controls 895)] were analysed. The EPIC samples were collected 2–5 years prior to diagnosis of breast or colorectal cancer. IGF2 DMR0 methylation levels in tumours were lower than matched non-tumour tissue. Hypomethylation of DMR0 was detected in breast (33%) and colorectal (80%) tumour tissues with a higher frequency than LOI indicating that methylation levels are a better indicator of cancer than LOI. In the EPIC population, the prevalence of IGF2 DMR0 hypomethylation was 9.5% and this correlated with increased age not cancer risk. Thus, IGF2 DMR0 hypomethylation occurs as an acquired tissue-specific somatic event rather than a constitutive innate epimutation. These results indicate that IGF2 DMR0 hypomethylation has diagnostic potential for colon cancer rather than value as a surrogate biomarker for constitutive LOI

Uncovering Genomic Regions Associated With 36 Agro-Morphological Traits in Indian Spring Wheat Using GWAS

Wheat genetic improvement by integration of advanced genomic technologies is one way of improving productivity. To facilitate the breeding of economically important traits in wheat, SNP loci and underlying candidate genes associated with the 36 agro-morphological traits were studied in a diverse panel of 404 genotypes. By using Breeders’ 35K Axiom array in a comprehensive genome-wide association study covering 4364.79 cM of the wheat genome and applying a compressed mixed linear model, a total of 146 SNPs (-log10P ≥ 4) were found associated with 23 traits out of 36 traits studied explaining 3.7–47.0% of phenotypic variance. To reveal this a subset of 260 genotypes was characterized phenotypically for six quantitative traits [days to heading (DTH), days to maturity (DTM), plant height (PH), spike length (SL), awn length (Awn_L), and leaf length (Leaf_L)] under five environments. Gene annotations mined ∼38 putative candidate genes which were confirmed using tissue and stage specific gene expression data from RNA Seq. We observed strong co-localized loci for four traits (glume pubescence, SL, PH, and awn color) on chromosome 1B (24.64 cM) annotated five putative candidate genes. This study led to the discovery of hitherto unreported loci for some less explored traits (such as leaf sheath wax, awn attitude, and glume pubescence) besides the refined chromosomal regions of known loci associated with the traits. This study provides valuable information of the genetic loci and their potential genes underlying the traits such as awn characters which are being considered as important contributors toward yield enhancement

Tumour specific CTCF point mutations abrogate CTCF function

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Feasibility and acceptability of ambulatory glucose profile in children with Type 1 diabetes mellitus: A pilot study

Background: Insulin administration and self-monitoring of blood glucose (SMBG) are pillars in the management of diabetes in children. Introduction of continuous glucose monitoring (CGM) has made it possible to understand the glycemic profiles which are not picked up by SMBG. Recent advent of flash glucose monitoring with inbuilt software to obtain ambulatory glucose profile (AGP) has emerged as a novel method to study glycemic patterns in adults with Type I diabetes. However, the use of AGP in children is yet to be explored. Methods: AGP was used in 46 children with Type 1 diabetes mellitus. Feasibility was measured regarding data and sensor failure. Acceptability was measured using a questionnaire. Results: Forty-six children (22 girls and 24 boys) with a mean age of 10.07 years and mean diabetes duration of 3.4 years were included in the study. In this cohort, for 30 (65.21%) subjects, the sensor remained in situ for a complete duration of 14 days. Except for minor discomfort, AGP was well accepted by most of the children and their parents. Conclusion: AGP is a feasible option for monitoring glycemic status in children with diabetes with a high rate of acceptance

Implementation of DDARC: Software Architecture for Debugging Distributed Programs

Debugging and testing is a larger part of the effort spent in a software development cycle. Debugging a program is time consuming and is a continuous cycle of code modification and testing. The task of debugging depends on the environment, the language, the operating system; the problem; and more interestingly even the individual program. We present a detailed implementation of software architecture for debugging distributed programs. The debugger based on the proposed architecture supports debugging based on a mechanism for controlling multiple processes, presenting the process states, grouping of processes for ease in controlling the distributed programs developed on PVM based computing paradigm

Development of Mucoadhesive Microsphere of Quinapril Hydrochloride for Treatment of Hypertension

Mucoadhesive drug delivery systems are very widely functional approach for delivery of system within the lumen of GIT to enhance drug absorption through the part of stomach with specific manner. Quinapril hydrochloride is the hydrochloride salt of quinapril, the ethyl ester of a non-sulfhydryl, angiotensin-converting enzyme (ACE) inhibitor, quinaprilat. The quinapril hydrochloride microspheres was prepared with a coat consisting of alginate polymer i.e. sodium alginate in combination of mucoadhesive polymer chitosan/ guargum by an ionic gelation process. The microspheres were evaluated for morphological character, particle size, micromeritic properties, percentage entrapment efficiency, in-vitro wash-off test and in-vitro release studies. The drug quinapril hydrochloride was study for the release over 24 h duration.&nbsp; The retarding nature of system maximizing the medication discharge rate at the appropriate site within specified time period for enhancing the bioavailability of drug at desired site of action to give successful treatment to the patients experiencing hypertension The drug release of the microspheres (QLMM1 – QLMM10) was slow, extended and dependent on the composition of galactomannan concentration of polymer and stirring speed during formulation used. The mucoadhesive microspheres were adhered at intestinal pH due to highly swelling nature of composition of polymers at this pH. So, increase the adhesive strength and retarded the drug release of best composition of CH:GG in the ratio of 1:3 (QLMM6). Guargum is a highly viscous material having a property of more swelling nature due to presence of galactommannan constituent. Thus, drug release from QLMM6 was slow and extended over a period of 24 h and these microcapsules were found suitable for oral controlled release formulations. Keywords: Gastro-retentive drug delivery, Mucoadhesive microsphere, Quinapril hydrochloride, Pulsatile drug delivery system, Treatment of hypertension, Natural polymer

Design, Optimization and Evaluation of Etoricoxib Castor Oil Emulgel for acute gout therapy

Etoricoxibis is a non-steroidal anti-inflammatory drug which showed anti-inflammatory, analgestic and antipyretic activities. Etoricxib is indicates in the management of Osteoarthritis, Rheumatoid and Acute gout. The major drawback of topical dosage form is dissolution and diffusion of drug in the delivery of hydrophobic drugs, and permeation through stratum corneum is for hydrophilic drugs, thus, to be referred as emulgels. In emulgel formulations, oil-in-water and water-in-oil emulsions are commonly utilised as vehicles to deliver different hydrophilic and hydrophobic medications to the skin. They're also good in dissolving drugs and penetrating skin. Oil-in-water emulsions are mostly used as medication bases that may be washed away. The proposed research project intends to create an emulgel containing the analgesic drug Etoricoxib. The created emulgel containing etoricoxib medicines has a topical route that is more permeable, regulated, and localised. The analytical procedure of etoricoxib medicine was assessed using described UV spectrophotometric techniques in pH Phosphate buffer 6.8 and exhibited good linearity of data, as demonstrated by correlation coefficients greater than 0.998. For organoleptic characteristics, flow qualities, and partitioning, preformulation experiments were employed. All result of preformulation study was satisfactory and drug was examined visually for their feeling after application on skin, color intensity, pH determination, consistency and extrudability determination. The result concluded that ETEG4 was best formulation. This formulation ETEG4 was prepared emulgel Carbopol 940 (2g), PVP, castor oil base. The drug release profile and release kinetics are two important characteristics of the dosage forms, which play an important role for describing dissolution profile of dosage form. The dissolution data was obtained and ETEG4 showed supercase II transport mechanism. Keywords: Etorcoxib, Castor Oil, Emulgel, Osteoarthritis, Rheumatoid, Acute gou

Land Cover Change Dynamics and their Impacts on Thermal Environment of Dadri Block, Gautam Budh Nagar, India

Land use / land cover (LULC) has been considered as one of the important bio-physical parameters and have significant affect on local environmental change, particularly increasing anthropogenic temperature. Remote sensing images from Landsat series satellites are a major information source for LULC change analysis. In the present investigation, long term changes in LULC and its negative impact on land surface temperature (LST) were analyzed using multi-temporal Landsat satellite images between 2000 to 2016. firstly LULC of the study area has been classified and temporal changes in land use classes were quantify, and observed that in most of the land use classes such as vegetation (-1.28 %), water bodies (-1.65 %), agriculture (-3.52) and open land (-2.43 %) have shown negative change, however large scale positive changes in built-up area (+8.87 %) has been observed during the analysis, which is mainly due to continuous urbanization and growth of population in the area. The classified thermal images from the same period also show mean temperature of the area has increased by 1.60 °C since last 16 years. The observation from the present study reveals that due to the large-scale land use change practices in urban and peri-urban area witnessed for the rising temperature due to loss natural vegetation and other natural resources