RESUMEN
Global investment in biomedical research has grown significantly over the last decades, reaching approximately a quarter of a trillion US dollars in 2010. However, not all of this investment is distributed evenly by gender. It follows, arguably, that scarce research resources may not be optimally invested (by either not supporting the best science or by failing to investigate topics that benefit women and men equitably). Women across the world tend to be significantly underrepresented in research both as researchers and research participants, receive less research funding, and appear less frequently than men as authors on research publications. There is also some evidence that women are relatively disadvantaged as the beneficiaries of research, in terms of its health, societal and economic impacts. Historical gender biases may have created a path dependency that means that the research system and the impacts of research are biased towards male researchers and male beneficiaries, making it inherently difficult (though not impossible) to eliminate gender bias. In this commentary, we - a group of scholars and practitioners from Africa, America, Asia and Europe - argue that gender-sensitive research impact assessment could become a force for good in moving science policy and practice towards gender equity. Research impact assessment is the multidisciplinary field of scientific inquiry that examines the research process to maximise scientific, societal and economic returns on investment in research. It encompasses many theoretical and methodological approaches that can be used to investigate gender bias and recommend actions for change to maximise research impact. We offer a set of recommendations to research funders, research institutions and research evaluators who conduct impact assessment on how to include and strengthen analysis of gender equity in research impact assessment and issue a global call for action.
Asunto(s)
Investigación Biomédica , Políticas , Sexismo , África , Américas , Asia , Europa (Continente) , Femenino , Identidad de Género , Política de Salud , Humanos , Masculino , Ciencia , Factores SexualesRESUMEN
The long-chain polyunsaturated n-6 and n-3 fatty acids are essential nutrients in membrane biogenesis and regulate gene expression via their eicosanoid metabolites. We investigated whether the n-6 and n-3 fatty acid supply as determined by maternal diet alters colonic phospholipid fatty acids, intestinal morphology, and epithelial barrier permeability during milk feeding with lasting effect on mucosal responsiveness to dinitrobenzene sulfonic acid (DNBS)-induced colitis in young adulthood. Female rats were fed diets with 20% energy from safflower oil (SO) or canola oil (CO), or 8% fish oil (FO) plus 2% SO (10% FO) or 18% FO plus 2% SO (20% FO) throughout gestation and lactation and offspring weaned to a standard diet at 21 days of age. At 15 days of age, pups in the 20% and 10% FO groups had lower 20:4n-6 and higher 20:5n-3 and 22:6n-3 in colon phospholipids (P < 0.01), shorter crypts (P < 0.05), and higher paracellular permeability than SO or CO groups. At 3 mo of age, male offspring in the FO groups showed lasting reduction of crypt depth and a heightened inflammatory response to DNBS. We demonstrate that early decreased colon 20:4n-6 with increased n-3 fatty acids impairs intestinal barrier development and sensitizes the colon response to inflammatory insults later in life.
Asunto(s)
Colitis/inducido químicamente , Grasas de la Dieta/farmacología , Intestinos/efectos de los fármacos , Intestinos/crecimiento & desarrollo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Colitis/fisiopatología , Colon/fisiología , Dinitrofluorobenceno/análogos & derivados , Dinitrofluorobenceno/toxicidad , Femenino , Masculino , Fenómenos Fisiologicos Nutricionales Maternos , Permeabilidad , Embarazo , Ratas , Ratas Sprague-DawleyRESUMEN
Hephaestin (Hp) is a membrane protein with ferroxidase activity that converts Fe(II) to Fe(III) during the absorption of nutritional iron in the gut. Using anti-peptide antibodies to predicted immunogenic regions of rodent Hp, previous immunocytochemical studies in rat, mouse, and human gut tissues localized Hp to the basolateral membranes of the duodenal enterocytes where the Hp was predicted to aid in the transfer of Fe(III) to transferrin in the blood. We used a recombinant soluble form of human Hp to obtain a high-titer polyclonal antibody to Hp. This antibody was used to identify the intracellular location of Hp in human gut tissue. Our immunocytochemical studies confirmed the previous localization of Hp in human enterocytes. However, we also localized Hp to the entire length of the gastrointestinal tract, the antral portion of the stomach, and to the enteric nervous system (both the myenteric and submucous plexi). Hp was also localized to human pancreatic beta-cells. In addition to its expression in the same cells as Hp, ferroportin was also localized to the ductal cells of the exocrine pancreas. The localization of the ferroxidase Hp to the neuronal plexi and the pancreatic beta cells suggests a role for the enzymatic function of Hp in the protection of these specialized cell types from oxidative damage.
Asunto(s)
Sistema Nervioso Entérico/metabolismo , Enterocitos/metabolismo , Tracto Gastrointestinal/metabolismo , Células Secretoras de Insulina/metabolismo , Proteínas de la Membrana/metabolismo , Antro Pilórico/metabolismo , Anticuerpos/inmunología , Especificidad de Anticuerpos/inmunología , Glándulas Duodenales/metabolismo , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Ceruloplasmina/inmunología , Duodeno/citología , Duodeno/metabolismo , Sistema Nervioso Entérico/citología , Células Epiteliales/metabolismo , Tracto Gastrointestinal/citología , Expresión Génica/genética , Humanos , Íleon/citología , Íleon/metabolismo , Insulina/metabolismo , Yeyuno/citología , Yeyuno/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/inmunología , Plexo Mientérico/citología , Plexo Mientérico/metabolismo , Neuronas/metabolismo , Páncreas/citología , Páncreas/metabolismo , Antro Pilórico/citología , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Plexo Submucoso/citología , Plexo Submucoso/metabolismoRESUMEN
Networks that represent connections between individuals can be valuable analytic tools. The Social Network Cytoscape app is capable of creating a visual summary of connected individuals automatically. It does this by representing relationships as networks where each node denotes an individual and an edge linking two individuals represents a connection. The app focuses on creating visual summaries of individuals connected by co-authorship links in academia, created from bibliographic databases like PubMed, Scopus and InCites. The resulting co-authorship networks can be visualized and analyzed to better understand collaborative research networks or to communicate the extent of collaboration and publication productivity among a group of researchers, like in a grant application or departmental review report. It can also be useful as a research tool to identify important research topics, researchers and papers in a subject area.
RESUMEN
It is well established that LH action is mediated primarily by adenylate cyclase/cAMP. However, the role of inositol phosphate/calcium in LH signaling is less well established. We examined the effects of gonadotropins in primary culture human granulosa-lutein cells and in HEK293 cells transiently transfected with human wild-type or chimeric gonadotropin receptors. The intracellular free calcium concentration was measured using fura-2 microspectrofluorometric techniques. Human (h) LH (2-4 microg/ml) and CG (10 IU/ml) consistently evoked oscillatory calcium signals in HEK293 cells transfected with hLH receptor, whereas hFSH (2-4 microg/ml) failed to elicit any response. Conversely, both hLH and hFSH failed to elicit a calcium response from HEK293 cells transfected with hFSHR, indicating the specificity of the response to the LH receptor. Pretreatment of transfected HEK293 cells with pertussis toxin (100 ng/ml) attenuated all gonadotropin-evoked calcium mobilization. Studies with chimeric LH receptor showed that the sequence of the long extracellular portion of the receptor was not critical for stimulation of PLC activity, but maintained agonist binding specificity. The C-terminal sequence of the receptor was clearly important for the generation of the basal calcium oscillations, but the precise extent of the critical sequence has yet to be identified. Although various subdivisions of this region were capable of stimulating calcium transients, an intact carboxyl-terminal third of the receptor was required for normal and sustained intracellular calcium signaling. Our study unequivocally shows that the hLH receptor is coupled to the inositol phosphate/calcium signaling pathway via a pertussis toxin-sensitive G protein-coupled receptor.
Asunto(s)
Calcio/metabolismo , Receptores de Gonadotropina/efectos de los fármacos , Cafeína/farmacología , Línea Celular , Gonadotropina Coriónica/farmacología , ADN Complementario/genética , Inhibidores Enzimáticos/farmacología , Femenino , Células de la Granulosa/efectos de los fármacos , Células de la Granulosa/metabolismo , Humanos , Indicadores y Reactivos , Luteína/metabolismo , Hormona Luteinizante/farmacología , Inhibidores de Fosfodiesterasa/farmacología , Proteínas Recombinantes de Fusión/metabolismo , Espectrometría de Fluorescencia , Relación Estructura-Actividad , Tapsigargina/farmacología , TransfecciónRESUMEN
In the stomach, the majority of substance P's effects are mediated by the activation of neurokinin-1 (NK1) receptors. The gastric cellular distribution of these receptors in Wistar and Sprague-Dawley rats was determined using immunocytochemistry. The localization of the NK1 receptors with respect to von Willebrand's factor, protein gene product 9.5, substance P, vasoactive intestinal peptide, and calcitonin gene-related peptide was also determined. Results show that NK1 receptor immunoreactivity was dependent on the duration of fixation. In corpus and antrum tissues that were fixed in 4% paraformaldehyde for 30 min, the presence of NK1 receptor immunoreactivity was demonstrated on nerve fibers throughout the stomach, on the surface and in the cytoplasm of myenteric cell bodies, on circular smooth muscle cells, and on vascular endothelial cells. This was observed in tissues from both rodent strains. Overnight fixation in the same fixative, however, demonstrated the presence of NK1 receptor immunoreactivity only on nerve fibers and cell bodies of the myenteric plexus, and on circular smooth muscle cells. In 30-min fixed tissues, the localization of NK1R immunoreactivity on vascular endothelial cells and nerve fibers was confirmed by co-localization with von Willebrand's factor and protein gene product 9.5 immunoreactivity, respectively. In both rodent strains, NK1 receptor immunoreactivity was co-localized with substance P immunoreactivity on nerve fibers of the longitudinal and circular muscle. In the Wistar rat, NK1 receptor immunoreactivity was co-localized with vasoactive intestinal peptide immunoreactivity or calcitonin gene-related peptide immunoreactivity throughout the stomach. However, in the Sprague-Dawley rat, NK1 receptor immunoreactivity was only co-localized with calcitonin gene-related peptide immunoreactivity in a minority of fibers of the circular muscle. The overall results of this study show that the antigenic epitopes of the NK1 receptor are sensitive to overfixation. When tissues were not overfixed, NK1 receptor immunoreactivity was distributed more extensively throughout the rat stomach than has been described previously. The results of this study provide the anatomical basis for many of the actions of substance P in the rat stomach.
Asunto(s)
Mucosa Gástrica/metabolismo , Receptores de Neuroquinina-1/metabolismo , Animales , Femenino , Inmunohistoquímica , Masculino , Ratas , Ratas Sprague-Dawley , Ratas Wistar , Coloración y Etiquetado , Estómago/citología , Sustancia P/metabolismo , Sinaptofisina/metabolismo , Tioléster Hidrolasas/metabolismo , Distribución Tisular , Fijación del Tejido , Ubiquitina Tiolesterasa , Péptido Intestinal Vasoactivo/metabolismo , Factor de von Willebrand/metabolismoAsunto(s)
Células Enteroendocrinas/citología , Células Epiteliales/citología , Mucosa Gástrica/citología , Antro Pilórico/citología , Células Cultivadas , Células Enteroendocrinas/microbiología , Células Epiteliales/microbiología , Mucosa Gástrica/microbiología , Helicobacter pylori/citología , Humanos , Microscopía Electrónica de Rastreo , Antro Pilórico/microbiología , Técnicas de Cultivo de TejidosRESUMEN
Somatostatin regulates multiple biological functions by acting through a family of five G protein-coupled receptors, somatostatin receptors (SSTRs) 1-5. Although all five receptor subtypes inhibit adenylate cyclase activity and decrease intracellular cAMP levels, specific receptor subtypes also couple to additional signaling pathways. In CCL39 fibroblasts expressing either human SSTR1 or SSTR2, we demonstrate that activation of SSTR1 (but not SSTR2) attenuated both thrombin- and integrin-stimulated Rho-GTP complex formation. The reduction in Rho-GTP formation in the presence of somatostatin was associated with decreased translocation of Rho and LIM kinase to the plasma membrane and fewer focal contacts. Activation of Rho resulted in the formation of intracellular actin stress fibers and cell migration. In CCL39-R1 cells, somatostatin treatment prevented actin stress fiber assembly and attenuated thrombin-stimulated cell migration through Transwell membranes to basal levels. To show that native SSTR1 shares the ability to inhibit Rho activation, we demonstrated that somatostatin treatment of human umbilical vein endothelial cells attenuated thrombin-stimulated Rho-GTP accumulation. These data show for the first time that a G protein-coupled receptor, SSTR1, inhibits the activation of Rho, the assembly of focal adhesions and actin stress fibers, and cell migration.
Asunto(s)
Actinas/metabolismo , Receptores de Somatostatina/metabolismo , Somatostatina/metabolismo , Fibras de Estrés/metabolismo , Proteínas de Unión al GTP rho/metabolismo , Adenilil Ciclasas/metabolismo , Animales , Línea Celular , Membrana Celular/metabolismo , Movimiento Celular , Cricetinae , AMP Cíclico/metabolismo , Endotelio Vascular/citología , Fibroblastos/metabolismo , Guanosina Trifosfato/metabolismo , Immunoblotting , Péptidos y Proteínas de Señalización Intracelular , Pulmón/citología , Fosforilación , Unión Proteica , Proteínas Serina-Treonina Quinasas/metabolismo , Transporte de Proteínas , Transducción de Señal , Trombina/metabolismo , Factores de Tiempo , Proteínas de Unión al GTP rac/metabolismo , Quinasas Asociadas a rhoRESUMEN
Little is known about the interactions between Helicobacter pylori, which specializes in colonizing the mucin layer that covers the gastric mucosa, and primary gastric epithelial cells. The expression pattern of Toll-like receptors (TLRs) in primary gastric epithelial cells and cell lines was compared. Primary cells did not express TLR4, whereas all cell lines expressed a nonsignaling form of TLR4. Because other cells within the mucosa expressed TLR4, it was next investigated whether H. pylori can be recognized by TLR4--they cannot. Moreover, H. pylori infection of primary cells induced a regulated production of interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha, whereas infection of cell lines only resulted in IL-8 production. The cytokine production in all cell types was strictly cag dependent. These findings indicate that, although the epithelium is important in directing the immune response against H. pylori infections, the response is independent of TLR4.
Asunto(s)
Proteínas de Drosophila , Mucosa Gástrica/microbiología , Helicobacter pylori/patogenicidad , Glicoproteínas de Membrana/metabolismo , Receptores de Superficie Celular/metabolismo , Adolescente , Adulto , Biopsia , Células Cultivadas , Células Epiteliales/inmunología , Células Epiteliales/microbiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Antro Pilórico/citología , Antro Pilórico/inmunología , Antro Pilórico/microbiología , Receptor Toll-Like 4 , Receptores Toll-Like , Células Tumorales CultivadasRESUMEN
The human gastric pathogen Helicobacter pylori attaches to antral epithelial cells in vivo. Cultured human antral epithelial cells, AGS and NCI-N87 cell lines, were grown in the absence or presence of H. pylori and compared with respect to gene transcript levels, protein expression, organization of the actin cytoskeleton, and the regulation of cell migration. The Clontech Neurobiology array detected differentially expressed transcripts, while Western blots were used to investigate related changes in protein levels. Infection with H. pylori consistently upregulated annexin II, S100 A7, Rho-GTP, and IQGAP-1, whereas SSTR-1 was downregulated upon H. pylori infection. In the adherens junction, E-cadherin and IQGAP-1 were translocated from the plasma membrane to intracellular vesicles. The primary and NCI-N87 cells were similar with respect to cell-cell and cell-matrix adhesion and cell migratory behavior; in contrast the AGS cells were significantly different from the primary gastric epithelial cell preparations, and thus caution must be used when using this cell line for studies of gastric disease. These studies demonstrate a correlation between H. pylori infection and alterations to epithelial cell adhesion molecules, including increased levels of Rho-GTP and cell migration. These data indicate that destabilizing epithelial cell adherence is one of the factors increasing the risk of H. pylori-infected individuals developing gastric cancer.
Asunto(s)
Uniones Adherentes/metabolismo , Movimiento Celular , Células Epiteliales/microbiología , Regulación de la Expresión Génica , Helicobacter pylori/patogenicidad , Proteínas/metabolismo , Adulto , Adhesión Bacteriana , Línea Celular , Células Cultivadas , Células Epiteliales/fisiología , Femenino , Perfilación de la Expresión Génica , Infecciones por Helicobacter/microbiología , Helicobacter pylori/fisiología , Humanos , Masculino , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Proteínas/genética , Antro Pilórico/citología , Antro Pilórico/microbiología , Receptores Acoplados a Proteínas G/metabolismo , VirulenciaRESUMEN
The incretins glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) are gut hormones that act via the enteroinsular axis to potentiate insulin secretion from the pancreas in a glucose-dependent manner. Both GLP-1 receptor and GIP receptor knockout mice (GLP-1R(-/-) and GIPR(-/-), respectively) have been generated to investigate the physiological importance of this axis. Although reduced GIP action is a component of type 2 diabetes, GIPR-deficient mice exhibit only moderately impaired glucose tolerance. The present study was directed at investigating possible compensatory mechanisms that take place within the enteroinsular axis in the absence of GIP action. Although serum total GLP-1 levels in GIPR knockout mice were unaltered, insulin responses to GLP-1 from pancreas perfusions and static islet incubations were significantly greater (40-60%) in GIPR(-/-) than in wild-type (GIPR(+/+)) mice. Furthermore, GLP-1-induced cAMP production was also elevated twofold in the islets of the knockout animals. Pancreatic insulin content and gene expression were reduced in GIPR(-/-) mice compared with GIPR(+/+) mice. Paradoxically, immunocytochemical studies showed a significant increase in beta-cell area in the GIPR-null mice but with less intense staining for insulin. In conclusion, GIPR(-/-) mice exhibit altered islet structure and topography and increased islet sensitivity to GLP-1 despite a decrease in pancreatic insulin content and gene expression.