Cloning and characterization of Enaptin, a novel giant actin-binding protein connecting the nucleus to the actin cytoskeleton

Summary ----------------------------------------------------------------------- The a-actinin superfamily is the largest of the F-actin cross-linking protein families Members of this superfamily have an actin-binding domain (ABD) consisting of a pair of calponin homology domains. We have cloned and characterized a novel giant actin-binding protein called Enaptin, which belongs to this family. Together with NUANCE, Enaptin forms a group of giant proteins that associate with the F-actin cytoskeleton as well as the nuclear membrane. Enaptin is composed of an N-terminal a-actinin type ABD, followed by a long coiled-coil rod and a transmembrane domain at the C-terminus. We have cloned and assembled a cDNA for Enaptin and found that the longest open reading frame of Enaptin encompasses 27,669 bp and predicts a 1014 kDa protein. The human Enaptin gene located on 6q23.1-25.3 spreads over 515 kb and gives rise to several splicing isoforms (Nesprin-1, Myne-1, Syne-1, CPG2). Northern blot analysis identified a >14 kb transcript and an additional transcript of 5.5 kb in brain. Using a polyclonal antibody against the ABD of Enaptin we detected a protein of approximately 400 kDa in tissues like brain and skeletal muscle. Further analysis showed that Enaptin is expressed in a wide range of tissues. Polyclonal antibodies generated against the C-terminus of Enaptin detected the protein at the nuclear envelope and in the cytosol of human fibroblasts. We showed that Enaptin is located in the outer nuclear membrane by selective permeabilisation of the plasma membrane with digitonin, furthermore its nuclear envelope localization was not affected by disruption of the actin cytoskeleton. Our studies also indicated that the nuclear envelope localization of Enaptin depended on lamin A/C. This is underlined by our findings in fibroblast cells from patients affected with laminopathies where we observed an altered distribution of Enaptin. With the N-terminal ABD domain and its C-terminal transmembrane domain, Enaptin has the potential to connect the nucleus to the actin cytoskeleton. Studies on Enaptin orthologues in lower eukaryotes proposed a role for these proteins in nuclear positioning and anchorage and in embryonic muscle development. Our myoblast differentiation studies showed an altered expression of Enaptin during differentiation. In order to reveal the functions of Enaptin and its link to human disease, we initiated a project for targeted disruption of Enaptin in the mouse

User Satisfaction on Library Resources and Services: A Case Study of Pontifical Institute of Theology and Philosophy Alwaye (PIA) Library at Mangalapuzha Campus

A case study was conducted to evaluate the users’ level of satisfaction with the Pontifical Institute of Theology and Philosophy, Alwaye (PIA) Library at Mangalapuzha Campus. The respondents were the resident seminarians of St. Joseph Pontifical Seminary, Alwaye and they belonged to the faculty of theology and philosophy. A total of 152 responses were received with a response rate of 86.86%. The study concluded that the overall satisfaction of respondents with the library resources and services was 76%. Results showed that the majority of the respondents satisfied with library timings and visited the library for the purpose of referring to books and journals. The reference section of the library was the most relied on the area and the encyclopedia collection was the most relied on resources. The results of this study would help both the PIA administration and St. Joseph Pontifical Seminary authority to take serious steps to strengthen and improve the resources and services

Diversity and Bioprospecting Potential of Bacteria Isolated from the Arctic: A Preliminary Study

Bioprospecting is a search for new or better bioproducts from biological sources preferably    from novel biodiversity. Molecules derived from natural products, particularly those produced by plants and microorganisms, have an excellent record of providing novel chemical structures for development as new pharmaceuticals. Bacteria living under stress are best sources for bioprospecting and also these organisms are least explored. This paper focuses on the bioprospecting and biotechnological potential of bacteria isolated from the Polar Regions. In this context, let us have a glance through the major cold adaptations of psychrophiles and their application potentials with special reference to psychrophilic enzymes. Our lab focus on identifying the biofilm inhibitors against bacterial pathogens and novel molecules of medical importance from bacteria and actinomycetes isolated from Arctic, the North Pole. We are also interested in profiling the diversity, multidrug resistance pattern   and the molecular mechanisms involved in bacteria isolated from the pristine environment

RhoJ/TCL Regulates Endothelial Motility and Tube Formation and Modulates Actomyosin Contractility and Focal Adhesion Numbers

Objective—RhoJ/TCL was identified by our group as an endothelial-expressed Rho GTPase. The aim of this study was to determine its tissue distribution, subcellular localization, and function in endothelial migration and tube formation. Methods and Results—Using in situ hybridization, RhoJ was localized to endothelial cells in a set of normal and cancerous tissues and in the vasculature of mouse embryos; endogenous RhoJ was localized to focal adhesions by immunofluorescence. The proangiogenic factor vascular endothelial growth factor activated RhoJ in endothelial cells. Using either small interfering (si)RNA-mediated knockdown of RhoJ expression or overexpression of constitutively active RhoJ (daRhoJ), RhoJ was found to positively regulate endothelial motility and tubule formation. Downregulating RhoJ expression increased focal adhesions and stress fibers in migrating cells, whereas daRhoJ overexpression resulted in the converse. RhoJ downregulation resulted in increased contraction of a collagen gel and increased phospho–myosin light chain, indicative of increased actomyosin contractility. Pharmacological inhibition of Rho-kinase (which phosphorylates myosin light chain) or nonmuscle myosin II reversed the defective tube formation and migration of RhoJ knockdown cells. Conclusion—RhoJ is endothelial-expressed in vivo, activated by vascular endothelial growth factor, localizes to focal adhesions, regulates endothelial cell migration and tube formation, and modulates actomyosin contractility and focal adhesion numbers

Utilization of various lignocellulosic biomass for the production of nanocellulose: a comparative study

Nanocellulose was successfully extracted from five different lignocellulosic biomass sources viz. banana rachis, sisal, kapok, pineapple leaf and coir using a combination of chemical treatments such as alkaline treatment, bleaching and acid hydrolysis. The shape, size and surface properties of the nanocellulose generally depend on the source and hydrolysis conditions. A comparative study of the fundamental properties of raw material, bleached and nanocellulose was carried out by means of Fourier transform infrared spectroscopy, scanning electron microscopy, atomic force microscopy, transmission electron microscopy, birefringence, X-ray diffraction, inverse gas chromatography and thermogravimetric analysis. Through the characterization of the nanocellulose obtained from different sources, the isolated nanocellulose showed an average diameter in the range of 10–25 nm, high crystallinity, high thermal stability and a great potential to be used with acid coupling agents due to a predominantly basic surface. This work provides an insight into the effective utilization of a variety of plant biomass as a potential source for nanocellulose extraction.info:eu-repo/semantics/publishedVersio

Mechanics and Pervaporation Performance of Ionic Liquid Modified CNT Based SBR Membranes - A Case Study for the Separation of Toluene/Heptane Mixtures

SBR/ionic liquid modified CNT Nanocomposites were prepared using different concentration of CNT. Nano scale dispersion of CNT affected the properties of SBR. The results have been compared with the unfilled SBR films. Cure characteristics and mechanical properties such as tensile strength, modulus, abrasion resistance and hardness were measured for different composites. Morphological behaviour and structural characteristics of the composites were investigated by AFM, TEM, FTIR and Raman spectroscopy. Microstructural development in presence of filler and interfacial interaction between modified CNT and polymer matrix led to enhanced properties. The pervaporation performances of membranes were analysed using a toluene/heptane mixture. Membranes displayed high selectivity towards heptane. The influence of feed composition on pervaporation was also analysed. The 5 phr CNT loaded membrane showed enhanced membrane permeance and selectivity value, an improvement of 18% over the neat polymer. A drop in selectivity and an increase in permeation rate were observed at higher CNT loadings

Data on cardiac lncRNA STX18-AS1 expression in developing human hearts and function during in vitro hESC-cardiomyocyte differentiation

This article presents data concerning STX18-AS1, a long noncoding RNA gene identified from a Genome-wide association study of Atrial Septal Defect (ASD). The data describes its expression patterns in human tissues and functions in regulating cardiomyocyte differentiation in vitro. STX18-AS1 is a lncRNA with a higher abundance in developing tissues, including hearts. Its transcription distribution within the embryonic hearts during key heart septation stages supports STX18-AS1’s association with risk SNPs for ASD. The CRISPR stem cell pool in which STX18-AS1 was knocked down, showed reduced CM differentiation efficiency and lower expression of key cardiac transcriptional factors. This indicated its regulative role in supporting the lineage specification from cardiac mesoderm into cardiac progenitors and cardiomyocytes. These data can benefit the understanding of human embryonic heart developmental biology, and the time-course changes of cardiac transcriptional factors during in vitro cardiomyocyte differentiation from human embryonic stem cells

Apelin Is Required for Non-Neovascular Remodeling in the Retina

Retinal pathologies are frequently accompanied by retinal vascular responses, including the formation of new vessels by angiogenesis (neovascularization). Pathological vascular changes may also include less well characterized traits of vascular remodeling that are non-neovascular, such as vessel pruning and the emergence of dilated and tortuous vessel phenotypes (telangiectasis). The molecular mechanisms underlying neovascular growth versus non-neovascular remodeling are poorly understood. We therefore undertook to identify novel regulators of non-neovascular remodeling in the retina by using the dystrophic Royal College of Surgeons (RCS) rat and the retinal dystrophy 1 (RD1) mouse, both of which display pronounced non-neovascular remodeling. Gene expression profiling of isolated retinal vessels from these mutant rodent models and wild-type controls revealed 60 differentially expressed genes. These included the genes for apelin (Apln) and for its receptor (Aplnr), both of which were strongly up-regulated in the mutants. Crossing RD1 mice into an Apln-null background substantially reduced vascular telangiectasia. In contrast, Apln gene deletion had no effect in two models of neovascular pathology [laser-induced choroidal neovascularization and the very low density lipoprotein receptor (Vldlr)-knockout mouse]. These findings suggest that in these models apelin has minimal effect on sprouting retinal angiogenesis, but contributes significantly to pathogenic non-neovascular remodeling

The urgent need for integrated science to fight COVID-19 pandemic and beyond

The COVID-19 pandemic has become the leading societal concern. The pandemic has shown that the public health concern is not only a medical problem, but also afects society as a whole; so, it has also become the leading scientifc concern. We discuss in this treatise the importance of bringing the world’s scientists together to fnd efective solu‑ tions for controlling the pandemic. By applying novel research frameworks, interdisciplinary collaboration promises to manage the pandemic’s consequences and prevent recurrences of similar pandemics

Intelligent Informatics : Proceedings of the International Symposium on Intelligent Informatics

This book constitutes the thoroughly refereed post-conference proceedings of the first International Symposium on Intelligent Informatics (ISI'12) held in Chennai, India during August 4-5, 2012. The 54 revised papers presented were carefully reviewed and selected from 165 initial submissions. The papers are organized in topical sections on data mining, clustering and intelligent information systems, multi agent systems, pattern recognition, signal and image processing and, computer networks and distributed systems. The book is directed to the researchers and scientists engaged in various fields of intelligent informatics