Hepatocyte growth factor and MET support mouse enteric nervous system development, the peristaltic response, and intestinal epithelial proliferation in response to injury

UNLABELLED: Factors providing trophic support to diverse enteric neuron subtypes remain poorly understood. We tested the hypothesis that hepatocyte growth factor (HGF) and the HGF receptor MET might support some types of enteric neurons. HGF and MET are expressed in fetal and adult enteric nervous system. In vitro, HGF increased enteric neuron differentiation and neurite length, but only if vanishingly small amounts (1 pg/ml) of glial cell line-derived neurotrophic factor were included in culture media. HGF effects were blocked by phosphatidylinositol-3 kinase inhibitor and by MET-blocking antibody. Both of these inhibitors and MEK inhibition reduced neurite length. In adult mice, MET was restricted to a subset of calcitonin gene-related peptide-immunoreactive (IR) myenteric plexus neurons thought to be intrinsic primary afferent neurons (IPANs). Conditional MET kinase domain inactivation (Met(fl/fl); Wnt1Cre+) caused a dramatic loss of myenteric plexus MET-IR neurites and 1-1\u27-dioctodecyl-3,3,3\u27,3\u27-tetramethylindocarbocyamine perchlorate (DiI) labeling suggested reduced MET-IR neurite length. In vitro, Met(fl/fl); Wnt1Cre+ mouse bowel had markedly reduced peristalsis in response to mucosal deformation, but normal response to radial muscle stretch. However, whole-bowel transit, small-bowel transit, and colonic-bead expulsion were normal in Met(fl/fl); Wnt1Cre+ mice. Finally, Met(fl/fl); Wnt1Cre+ mice had more bowel injury and reduced epithelial cell proliferation compared with WT animals after dextran sodium sulfate treatment. These results suggest that HGF/MET signaling is important for development and function of a subset IPANs and that these cells regulate intestinal motility and epithelial cell proliferation in response to bowel injury. SIGNIFICANCE STATEMENT: The enteric nervous system has many neuronal subtypes that coordinate and control intestinal activity. Trophic factors that support these neuron types and enhance neurite growth after fetal development are not well understood. We show that a subset of adult calcitonin gene-related peptide (CGRP)-expressing myenteric neurons produce MET, the receptor for hepatocyte growth factor, and that loss of MET activity affects peristalsis in response to mucosal stroking, reduces MET-immunoreactive neurites, and increases susceptibility to dextran sodium sulfate-induced bowel injury. These observations may be relevant for understanding and treating intestinal motility disorders and also suggest that enhancing the activity of MET-expressing CGRP neurons might be a useful strategy to reduce bowel inflammation

Revaluación voluntaria de los activos fijos y su incidencia en el estado de situación financiera, y el estado de cambios en el patrimonio en la empresa SCHEMIN PERÚ S.A.

El presente trabajo Revaluación Voluntaria de los Activos Fijos y su Incidencia en el Estado de Situación Financiera, y el Estado de Cambios en el Patrimonio en la empresa Schemin Perú S.A, está orientada a determinar como la aplicación de la revaluación voluntaria en los activos fijos en la empresa Schemin Perú S.A. Permitirá disponer con información actualizada y precisa, para una toma de decisiones más certeras y de acuerdo a sus objetivos estratégicos empresariales; demostrando que una revaluación voluntaria no es simplemente un acto contable, sino que implica una herramienta eficaz para el empresario. La investigación se realizó a través del análisis de los estados financieros y del libro de activos fijos de los años 2015-2016 de la empresa Schemin Perú S.A, obteniendo como resultado activos totalmente depreciados o próximos a estarlo debido a que la vida útil fue estimada de acuerdo al método de línea recta y de porcentajes conforme a la Ley de Impuesto a la Renta (LIR); sin embargo estos seguían siendo utilizados para su trabajo habitual. El informe pericial elaborado a los activos fijos refleja que se encuentran en buenas condiciones para su uso. Con los nuevos datos se tomó la iniciativa de elaborar la Revaluación voluntaria de los Activos fijos, donde se muestra una variación en los Estados de Situación Financiera al 31 de diciembre del 2016. La incidencia en el Estado de Situación Financiera es de S/ 306,521.61 soles y en el Estado de Cambios en el Patrimonio es de S/ 327,079.09 solesThe present work; voluntary revaluation of fixed assets and its incidence in the state of financial position and the state of changes in the equity of Schemin Peru S.A. company, is oriented to determine how the application of voluntary revaluation in the fixed assets of the Schemin Peru S.A. company will provide updated and precise information in order to improve the decision making of the company considering its business strategical objectives. This will show that a voluntary revaluation is not just an accounting act, but also an effective tool for the businessman. The research was done through the analysis of the financial statements and the fixed assets book from Schemin Peru S.A. of the 2015 – 2016 periods, resulting in assets totally or almost despised. This happens because the useful life was estimated in terms of the straight line method and percentages according to taxes laws of our country; however, these still have been used in its working routine. The expert report elaborated to the fixed assets reflects that it is in good conditions for its use. With the new data, we elaborated the voluntary revaluation of fixed assets, in which it shows a variation in the statements of financial situation as of December 31, 2016 The incidence in the statement of financial situation is S/. 306,521.61 and in the statement of changes in equity is S/. 327,079.09.Trabajo de investigació

Ibuprofen slows migration and inhibits bowel colonization by enteric nervous system precursors in zebrafish, chick and mouse

AbstractHirschsprung Disease (HSCR) is a potentially deadly birth defect characterized by the absence of the enteric nervous system (ENS) in distal bowel. Although HSCR has clear genetic causes, no HSCR-associated mutation is 100% penetrant, suggesting gene–gene and gene-environment interactions determine HSCR occurrence. To test the hypothesis that certain medicines might alter HSCR risk we treated zebrafish with medications commonly used during early human pregnancy and discovered that ibuprofen caused HSCR-like absence of enteric neurons in distal bowel. Using fetal CF-1 mouse gut slice cultures, we found that ibuprofen treated enteric neural crest-derived cells (ENCDC) had reduced migration, fewer lamellipodia and lower levels of active RAC1/CDC42. Additionally, inhibiting ROCK, a RHOA effector and known RAC1 antagonist, reversed ibuprofen effects on migrating mouse ENCDC in culture. Ibuprofen also inhibited colonization of Ret+/− mouse bowel by ENCDC in vivo and dramatically reduced bowel colonization by chick ENCDC in culture. Interestingly, ibuprofen did not affect ENCDC migration until after at least three hours of exposure. Furthermore, mice deficient in Ptgs1 (COX 1) and Ptgs2 (COX 2) had normal bowel colonization by ENCDC and normal ENCDC migration in vitro suggesting COX-independent effects. Consistent with selective and strain specific effects on ENCDC, ibuprofen did not affect migration of gut mesenchymal cells, NIH3T3, or WT C57BL/6 ENCDC, and did not affect dorsal root ganglion cell precursor migration in zebrafish. Thus, ibuprofen inhibits ENCDC migration in vitro and bowel colonization by ENCDC in vivo in zebrafish, mouse and chick, but there are cell type and strain specific responses. These data raise concern that ibuprofen may increase Hirschsprung disease risk in some genetically susceptible children

KRAS-specific inhibition using a DARPin binding to a site in the allosteric lobe.

Inhibiting the RAS oncogenic protein has largely been through targeting the switch regions that interact with signalling effector proteins. Here, we report designed ankyrin repeat proteins (DARPins) macromolecules that specifically inhibit the KRAS isoform by binding to an allosteric site encompassing the region around KRAS-specific residue histidine 95 at the helix α3/loop 7/helix α4 interface. We show that these DARPins specifically inhibit KRAS/effector interactions and the dependent downstream signalling pathways in cancer cells. Binding by the DARPins at that region influences KRAS/effector interactions in different ways, including KRAS nucleotide exchange and inhibiting KRAS dimerization at the plasma membrane. These results highlight the importance of targeting the α3/loop 7/α4 interface, a previously untargeted site in RAS, for specifically inhibiting KRAS function

The Voicing Hidden Histories Project: Participatory Video in Development, Soft Power and Film Language

In this article, we wish to reflect upon some of the findings of a recent Arts and Humanities Research Council-funded Global Challenges Research Fund Participatory Video (PV) project ‘Voicing Hidden Histories’. Working in South Africa, India and Brazil, this project has been using PV to support specific marginalised communities in each country to challenge the way their nations present themselves – and in particular their national history – to the world via ‘nation branding’ and other ‘soft power’ initiatives. Specifically, we ask: why use filmmaking as an international development tool? What are the enablers of – and barriers to – successful PV initiatives and what does ‘success’ mean in this context? Moreover, while such projects invariably make claims for PV as a particularly effective method for ‘giving’ communities ‘voice’ – however such potentially patronising terms might be defined – very little space is usually dedicated to the exploration of the films produced in such projects, that is to the specific articulation of this ‘voice’. Thus, we also wish to challenge a trend in the analysis of such projects that focuses entirely on questions of methodology and an understanding of PV as a process, largely ignoring the products made

Surfaceome interrogation using an RNA-seq approach highlights leukemia initiating cell biomarkers in an LMO2 T cell transgenic model.

The surfaceome is critical because surface proteins provide a gateway for internal signals and transfer of molecules into cells, and surfaceome differences can influence therapy response. We have used a surfaceome analysis method, based on comparing RNA-seq data between normal and abnormal cells (Surfaceome DataBase Mining or Surfaceome DBM), to identify sets of upregulated cell surface protein mRNAs in an LMO2-mediated T-ALL mouse model and corroborated by protein detection using antibodies. In this model the leukemia initiating cells (LICs) comprise pre-leukaemic, differentiation inhibited thymocytes allowing us to provide a profile of the LIC surfaceome in which GPR56, CD53 and CD59a are co-expressed with CD25. Implementation of cell surface interaction assays demonstrates fluid interaction of surface proteins and CD25 is only internalized when co-localized with other proteins. The Surfaceome DBM approach to analyse cancer cell surfaceomes is a way to find targetable surface biomarkers for clinical conditions where RNA-seq data from normal and abnormal cell are available