Structural Insights into the Shroom-Rock Interaction and the Regulation of the Actomyosin Cytoskeleton

During development, tissues undergo precise and controlled changes in shape and size. Various signaling pathways regulate these changes, temporally and spatially, by altering cytoskeleton dynamics to alter cell shape. One such pathway involves the Shroom and Rock proteins that reorganize the actomyosin cytoskeleton in epithelial cells to alter tissue morphology. The Shroom family of proteins are multi-domain, actin-binding proteins required for many developmental processes such as neural tube formation, and retinal morphogenesis. Shroom proteins interact with Rho-kinase (Rock), another conserved cytoskeleton regulator, to activate non-muscle Myosin II and assemble a contractile actomyosin network. All Shroom proteins contain a highly conserved C-terminal domain called Shroom Domain 2 (SD2) that interacts with the Rock Shroom binding domain (SBD) and is required for Shroom-mediated apical constriction. In the Shroom-Rock system it is unclear how this interaction activates the kinase activity of Rock. The goal of this dissertation is to understand the mechanistic details of the Shroom-Rock interaction. Using structural studies I have started to dissect the SD2-SBD interaction. I first determined the crystal structure of the Drosophila Shroom SD2 domain at 2.7 Å resolution to be a novel fold composed of a three-segmented, anti-parallel, coiled-coil dimer. Using mutational analysis and a combination of in vivo and in vitro assays we identified surfaces within the central coiled-coil segment of the SD2 domain that mediate Rock binding. The anti-parallel nature of the SD2 domain introduces internal symmetry into the SD2 domain such that there are two identical binding sites for Rock on opposite sides of the molecules suggesting interesting implications for the Shroom-Rock interaction. We also determined the crystal structure of the Rock SBD to 2.5 Å resolution and saw that it is a parallel coiled-coil dimer. Using mutational analysis combined with biochemical assays I have identified two conserved patches on opposite ends of Rock SBD that are required for Shroom interaction. These patches, unlike the patches on the SD2 domain, are not identical. Biochemical characterization of the SD2-SBD complex suggests that molar ratio of this complex is 1:1. Based on these results we can start to suggest models for how Shroom and Rock interact

Energetic Water Quality Analysis Auditing Scheme With IoT Environment

In nowadays worldwide, Internet of Things (IoT) and Remote Sensing (RS) approaches are being used in special areas of study for surveillance, generating and also assessing statistics from remote locations. Consuming water is a totally important product for all humans as alcohol consumption water energies encounter lots of brand-new requiring situations in real-time procedure. These frustrating situations stem because of constricted water resources, growing populace, aging framework and also many others. Consequently, there is a need for higher methodologies to reveal the water premium. In order to make certain the secure supply of consuming water the fantastic needs to be checked in real-time. In this paper, we mean to supply the layout as well as enhancement of a low-fee tool for real monitoring of water fantastic in IoT surroundings. The criteria together with temperature, PH, turbidity, float sensor of the water may be determined. The measured values from the sensing units can be refined by means of the centre controller. The ARM 7 version may be utilized as a centre controller. Finally, the sensor truths may be appeared on the internet making use of WI-FI equipment

In-orbit Performance of UVIT on ASTROSAT

We present the in-orbit performance and the first results from the ultra-violet Imaging telescope (UVIT) on ASTROSAT. UVIT consists of two identical 38cm coaligned telescopes, one for the FUV channel (130-180nm) and the other for the NUV (200-300nm) and VIS (320-550nm) channels, with a field of view of 28 $arcmin$. The FUV and the NUV detectors are operated in the high gain photon counting mode whereas the VIS detector is operated in the low gain integration mode. The FUV and NUV channels have filters and gratings, whereas the VIS channel has filters. The ASTROSAT was launched on 28th September 2015. The performance verification of UVIT was carried out after the opening of the UVIT doors on 30th November 2015, till the end of March 2016 within the allotted time of 50 days for calibration. All the on-board systems were found to be working satisfactorily. During the PV phase, the UVIT observed several calibration sources to characterise the instrument and a few objects to demonstrate the capability of the UVIT. The resolution of the UVIT was found to be about 1.4 - 1.7 $arcsec$ in the FUV and NUV. The sensitivity in various filters were calibrated using standard stars (white dwarfs), to estimate the zero-point magnitudes as well as the flux conversion factor. The gratings were also calibrated to estimate their resolution as well as effective area. The sensitivity of the filters were found to be reduced up to 15\% with respect to the ground calibrations. The sensitivity variation is monitored on a monthly basis. UVIT is all set to roll out science results with its imaging capability with good resolution and large field of view, capability to sample the UV spectral region using different filters and capability to perform variability studies in the UV.Comment: 10 pages, To appear in SPIE conference proceedings, SPIE conference paper, 201

Structure of a highly conserved domain of rock1 required for shroom-mediated regulation of cell morphology

Rho-associated coiled coil containing protein kinase (Rho-kinase or Rock) is a well-defined determinant of actin organization and dynamics in most animal cells characterized to date. One of the primary effectors of Rock is non-muscle myosin II. Activation of Rock results in increased contractility of myosin II and subsequent changes in actin architecture and cell morphology. The regulation of Rock is thought to occur via autoinhibition of the kinase domain via intramolecular interactions between the N-terminus and the C-terminus of the kinase. This autoinhibited state can be relieved via proteolytic cleavage, binding of lipids to a Pleckstrin Homology domain near the C-terminus, or binding of GTP-bound RhoA to the central coiled-coil region of Rock. Recent work has identified the Shroom family of proteins as an additional regulator of Rock either at the level of cellular distribution or catalytic activity or both. The Shroom-Rock complex is conserved in most animals and is essential for the formation of the neural tube, eye, and gut in vertebrates. To address the mechanism by which Shroom and Rock interact, we have solved the structure of the coiled-coil region of Rock that binds to Shroom proteins. Consistent with other observations, the Shroom binding domain is a parallel coiled-coil dimer. Using biochemical approaches, we have identified a large patch of residues that contribute to Shrm binding. Their orientation suggests that there may be two independent Shrm binding sites on opposing faces of the coiled-coil region of Rock. Finally, we show that the binding surface is essential for Rock colocalization with Shroom and for Shroom-mediated changes in cell morphology. © 2013 Mohan et al

Instructional Models for Course-Based Research Experience (CRE) Teaching

The course-based research experience (CRE) with its documented educational benefits is increasingly being implemented in science, technology, engineering, and mathematics education. This article reports on a study that was done over a period of 3 years to explicate the instructional processes involved in teaching an undergraduate CRE. One hundred and two instructors from the established and large multi-institutional SEA-PHAGES program were surveyed for their understanding of the aims and practices of CRE teaching. This was followed by large-scale feedback sessions with the cohort of instructors at the annual SEA Faculty Meeting and subsequently with a small focus group of expert CRE instructors. Using a qualitative content analysis approach, the survey data were analyzed for the aims of inquiry instruction and pedagogical practices used to achieve these goals. The results characterize CRE inquiry teaching as involving three instructional models: 1) being a scientist and generating data; 2) teaching procedural knowledge; and 3) fostering project ownership. Each of these models is explicated and visualized in terms of the specific pedagogical practices and their relationships. The models present a complex picture of the ways in which CRE instruction is conducted on a daily basis and can inform instructors and institutions new to CRE teaching

Usefulness of The Reflux Symptom Index in the Management of Laryngopharyngeal Reflux

Introduction Laryngopharyngeal Reflux (LPR) is highly prevalent in the general population and its impact on health systems is growing dramatically by the day. The contents of the stomach flowing back into the oesophagus, pharynx and larynx because of a transient relaxation of the lower oesophageal sphincter leads to a spectrum of symptoms diagnosed as LPR and Gastroesophageal Reflux Disease (GERD). The aim was to study in detail the symptoms of LPR and to ascertain if and how they hamper the routine of an individual by using the Reflux Symptom Index (RSI).  Materials and Methods The Reflux Symptom Index (RSI) is a self-administered nine-item outcomes instrument for LPR.  Ninety-one patients with clinically diagnosed LPR were taken up for this study and were issued the RSI (translated into the local language for better results) before and after treatment.  Data was assessed at the end of 3 months and 6 months.  Results The tabulated data showed significant improvement in the symptomatic index after treatment.   Conclusion It can thus be concluded that RSI is still highly valid in the follow-up for patients with LPR because it can be easily administered and gives accurate results with excellent validation

Crystal structure of 2-phenylethanaminium 3-carboxyprop-2-enoate

The title molecular salt, C8H12N+·C4H3O4−, crystallized with two independent cations and anions in the asymmetric unit. The ethanaminium side chains of the cations exhibit anti conformations [C—C—C—N torsion angles = 176.5 (3) and −179.4 (3)°]. In the crystal, N—H...O and C—H...O hydrogen bonds connect adjacent anions and cations, and , O—H...O hydrogen bonds connect adjacent anions, generating sheets parallel to (001)

Crystal structure of 2-phenylethylaminium 4-nitrophenolate monohydrate

In the title hydrated molecular salt, C8H12N+·C6H4NO3−·H2O, the conformation of the side chain in the cation is anti [C—C—C—N = 179.62 (12)°] and the dihedral angle between the aromatic ring and the nitro group in the anion is 3.34 (11)°. In the crystal, the components are linked by O—H...O and N—H...O hydrogen bonds, generating (10-1) sheets, which feature R44(21) loops. The sheets interact by weak aromatic π–π stacking interactions [centroid–centroid distance = 3.896 (3) Å], forming a three-dimensional network