RESUMEN
Understanding the spatial dynamics and drivers of wildlife pathogens is constrained by sampling logistics, with implications for advancing the field of landscape epidemiology and targeted allocation of management resources. However, visually apparent wildlife diseases, when combined with remote-surveillance and distribution modelling technologies, present an opportunity to overcome this landscape-scale problem. Here, we investigated dynamics and drivers of landscape-scale wildlife disease, using clinical signs of sarcoptic mange (caused by Sarcoptes scabiei) in its bare-nosed wombat (BNW; Vombatus ursinus) host. We used 53,089 camera-trap observations from over 3261 locations across the 68,401 km2 area of Tasmania, Australia, combined with landscape data and ensemble species distribution modelling (SDM). We investigated: (1) landscape variables predicted to drive habitat suitability of the host; (2) host and landscape variables associated with clinical signs of disease in the host; and (3) predicted locations and environmental conditions at greatest risk of disease occurrence, including some Bass Strait islands where BNW translocations are proposed. We showed that the Tasmanian landscape, and ecosystems therein, are nearly ubiquitously suited to BNWs. Only high mean annual precipitation reduced habitat suitability for the host. In contrast, clinical signs of sarcoptic mange disease in BNWs were widespread, but heterogeneously distributed across the landscape. Mange (which is environmentally transmitted in BNWs) was most likely to be observed in areas of increased host habitat suitability, lower annual precipitation, near sources of freshwater and where topographic roughness was minimal (e.g. human modified landscapes, such as farmland and intensive land-use areas, shrub and grass lands). Thus, a confluence of host, environmental and anthropogenic variables appear to influence the risk of environmental transmission of S. scabiei. We identified that the Bass Strait Islands are highly suitable for BNWs and predicted a mix of high and low suitability for the pathogen. This study is the largest spatial assessment of sarcoptic mange in any host species, and advances understanding of the landscape epidemiology of environmentally transmitted S. scabiei. This research illustrates how host-pathogen co-suitability can be useful for allocating management resources in the landscape.
Asunto(s)
Marsupiales , Escabiosis , Animales , Humanos , Escabiosis/epidemiología , Efectos Antropogénicos , Ecosistema , Sarcoptes scabiei , Animales SalvajesRESUMEN
Fast purinergic signaling is mediated by ATP and ATP-gated ionotropic P2X receptors (P2XRs), and it is implicated in pain-related behaviors. The properties exhibited by P2XRs vary between those expressed in heterologous cells and in vivo. Several modulators of ligand-gated ion channels have recently been identified, suggesting that there are P2XR functional modulators in vivo. Here, we establish a genome-wide open reading frame (ORF) collection and perform functional screening to identify modulators of P2XR activity. We identify TMEM163, which specifically modulates the channel properties and pharmacology of P2XRs. We also find that TMEM163 is required for full function of the neuronal P2XR and a pain-related ATP-evoked behavior. These results establish TMEM163 as a critical modulator of P2XRs in vivo and a potential target for the discovery of drugs for treating pain.
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Adenosina Trifosfato/farmacología , Conducta Animal/efectos de los fármacos , Proteínas de la Membrana/metabolismo , Receptores Purinérgicos P2X/metabolismo , Animales , Calcio/metabolismo , Potenciales Evocados/efectos de los fármacos , Femenino , Genoma , Células HEK293 , Humanos , Proteínas de la Membrana/antagonistas & inhibidores , Proteínas de la Membrana/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuronas/metabolismo , Sistemas de Lectura Abierta/genética , Dolor/patología , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Receptores Purinérgicos P2X/genética , Receptores Purinérgicos P2X3/deficiencia , Receptores Purinérgicos P2X3/genética , Receptores Purinérgicos P2X3/metabolismoRESUMEN
Accurate and reliable quantification methods for gluten in food are necessary to ensure proper product labeling and thus safeguard the gluten sensitive consumer against exposure. Immunochemical detection is the method of choice, as it is sensitive, rapid and relatively easy to use. Although a wide range of detection kits are commercially available, there are still many difficulties in gluten detection that have not yet been overcome. This review gives an overview of the currently commercially available immunochemical detection methods, and discusses the problems that still exist in gluten detection in food. The largest problems are encountered in the extraction of gluten from food matrices, the choice of epitopes targeted by the detection method, and the use of a standardized reference material. By comparing the available techniques with the unmet needs in gluten detection, the possible benefit of a new multiplex immunoassay is investigated. This detection method would allow for the detection and quantification of multiple harmful gluten peptides at once and would, therefore, be a logical advancement in gluten detection in food.
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Glútenes/análisis , Inmunoensayo/métodos , Enfermedad Celíaca/dietoterapia , Enfermedad Celíaca/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Epítopos/inmunología , Análisis de los Alimentos/métodos , Etiquetado de Alimentos , Glútenes/inmunología , Humanos , Péptidos/análisis , Péptidos/inmunología , Triticum/química , Triticum/inmunologíaRESUMEN
Amylose is able to form helical inclusion complexes with lysophosphatidylcholine (LPC). This complexation influences the functional and rheological properties of wheat starch; however it is well known that the formation of these complexes lead the starchy systems to a slower enzymatic hydrolysis. Based on this, to benefit from both the structuring properties of starch and also lower digestibility of the inclusion complexes, the objective of this study is the formation of amylose-LPC inclusion complexes while developing a firm network providing the desired functional properties in a starchy system. To investigate the influence of amylose-LPC complex formation at different stages of starch gelation on the viscosity behavior of wheat starch, 3% (w/w) LPC was added at three different points of the viscosity profile, obtained by rapid visco analyzer (RVA). LPC addition at all points affected the gelation behavior of wheat starch as compared with the reference. LPC addition at half-peak and peak of the viscosity profile resulted in a viscosity increase during cooling. Measuring the dynamic rheological properties of the freshly prepared gelatinized samples showed a decrease of storage modulus (G') and loss modulus (G") in the presence of LPC. During storage, in the presence of LPC, a lower elasticity was observed which indicates a lower rate of amylose retrogradation due to complexation with LPC.
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Amilosa/metabolismo , Lisofosfatidilcolinas/metabolismo , Reología , Almidón/química , Triticum/química , Amilosa/química , Elasticidad , Geles , Hidrólisis , Lisofosfatidilcolinas/química , Temperatura , ViscosidadRESUMEN
Anorexia nervosa (AN) is a serious, potentially life-threatening disorder characterized by severe weight loss, dysregulated eating, and often excessive exercise. While psychiatric illnesses such as depression are associated with increased levels of pro-inflammatory mediators, evidence for such disturbances in patients with AN has been less clear. In an exploratory study of possible disturbances in immune responses in AN, we assayed a panel of cytokines and chemokines in the blood of patients undergoing inpatient treatment, testing the hypothesis that metabolic disturbances in this disease would lead to a pattern of immune disturbances distinct from that of other psychiatric diseases. For this purpose, we evaluated patients by the Beck Depression Inventory-II (BDI-II) and the Eating Disorders Examination-Questionnaire and assessed cytokines and chemokines by enzyme-linked immunosorbent assays. Patients reported a moderate level of depression (mean BDI-II = 22.6) but exhibited few immunologic abnormalities of the kind associated with major depressive disorder [e.g., increased interleukin (IL)-6]; RANTES showed the most frequent elevations and was increased in 4 of the patients studied. Together, these findings suggest that features of AN such as loss of adipose tissue and excessive exercise may attenuate cytokine production and thus modulate the experience of illness that impacts on core features of disease.
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Anorexia Nerviosa/sangre , Quimiocina CCL5/sangre , Depresión/complicaciones , Interleucina-6/sangre , Adolescente , Adulto , Composición Corporal , Quimiocina CCL5/biosíntesis , Ensayo de Inmunoadsorción Enzimática , Ejercicio Físico , Femenino , Humanos , Persona de Mediana Edad , Proyectos Piloto , Escalas de Valoración Psiquiátrica , Encuestas y Cuestionarios , Pérdida de Peso , Adulto JovenRESUMEN
Amylose forms inclusion complexes with lysophosphatidylcholine (LPC), that decrease the susceptibility of amylose to amylase degradation. This study on the influence of complexation on starch susceptibility to amylase explains the nature of this protective effect. Wheat starch suspensions (9% w/w) containing 0.5-5% LPC were subjected to hydrolysis by porcine pancreatic α-amylase at 37 °C for several digestion times. The digesta were analysed by size-exclusion chromatography (SEC). The molar mass distribution was closely dependent on the digestion time and amount of LPC. This study precisely demonstrates the alteration of the digestion profile of starch on a molecular level, influenced by amylose-LPC complexation; however the effect depends on the digestion time. During 15 and 30 min digestion, inclusion complexes not only protect amylopectin in the initial hydrolysis stage, but also demonstrate lower susceptibility of the molecular amylose complexes to amylase hydrolysis. Digestion for 240 min resulted in a lower oligosaccharide peak concentration, in the presence of a high LPC concentration, which is related to less degradation of complexed amylose fraction.
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Amilosa/metabolismo , Digestión , Lisofosfatidilcolinas/metabolismo , Almidón/metabolismo , Triticum/metabolismo , alfa-Amilasas/metabolismo , Amilosa/química , Animales , Cromatografía en Gel , Hidrólisis , Lisofosfatidilcolinas/química , Modelos Biológicos , Peso Molecular , Almidón/química , Porcinos , Triticum/química , alfa-Amilasas/químicaRESUMEN
This study was aimed to assess the role of lysophosphatidylcholine (LPC) in the development of slowly digestible starch (SDS). The influence of LPC, on the enzymatic degradation of diluted 9% wheat starch suspensions (w/w) was investigated, using an in vitro digestion method. Wheat starch suspensions containing 0.5-5% LPC (based on starch) were heated in a Rapid Visco Analyser (RVA) till 95 °C and subjected to enzyme hydrolysis by porcine pancreatic α-amylase at 37 °C for several digestion periods. In vitro digestion measurements demonstrated that complexing starch with 5% LPC leads to a 22% decrease in rate of reducing sugar compared to the reference while the samples containing 0.5% LPC showed an equal digestibility comparable to the control. A clear decrease in the formation of reducing sugars was observed in presence of 2-5% LPC, since the results after 15 min digestion imply the formation of SDS due to the formation of amylose-LPC inclusion complexes. The DSC measurements proved the presence of amylose-LPC inclusion complexes even after 240 min digestion demonstrating the low susceptibility of amylose-V complexes to amylase.
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Amilosa/metabolismo , Lisofosfatidilcolinas/metabolismo , Almidón/metabolismo , Triticum/metabolismo , alfa-Amilasas/metabolismo , Animales , Sus scrofa , Temperatura , Factores de Tiempo , ViscosidadRESUMEN
Starch is an omnipresent constituent which is used for its nutritional and structuring properties. Recently concerns have been raised since starch is a source of readily available glucose which is tightly correlated with diabetes type II and obesity. For this reason, the possibilities for modulating the digestibility of starch while preserving its functional properties were investigated; therefore the focus of this paper is on starch gelatinization and the effect of lysophosphatidylcholine (LPC) on the structuring properties of wheat starch. The effect of LPC on thermal properties and viscosity behavior of starch suspensions was studied using DSC and RVA, respectively. The influence on granular structure was observed by light microscopy. The RVA profile demonstrated no viscosity increase at high LPC concentrations which proves intact granular structure after gelatinization. LPC in intermediate concentrations resulted in a notable delay of pasting; however the peak and end viscosities were influenced as well. Lower LPC concentrations demonstrated a higher peak viscosity as compared with pure starch suspensions. DSC results imply that inclusion complexes of amylose-LPC might be formed during pasting time. Since the viscosity profiles are changed by LPC addition, swelling power and solubility of starch granules are influenced as well. LPC hinders swelling power and solubility of starch granules which are stimulated by heating.
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Lisofosfatidilcolinas/química , Almidón/química , Triticum/química , Amilosa/química , Animales , Yema de Huevo/química , Geles/química , Solubilidad , Coloración y Etiquetado , Propiedades de Superficie , Suspensiones/química , Termodinámica , Factores de Tiempo , ViscosidadRESUMEN
BACKGROUND: We internally validated previously published rates of remission, continuation and incidence of broadly defined eating disorders during pregnancy in the Norwegian Mother and Child Cohort (MoBa) at the Norwegian Institute of Public Health. METHOD: A total of 77 267 pregnant women enrolled at 17 weeks gestation between 2001 and 2009 were split into a training sample (n = 41 243) from the version 2 dataset and a validation sample (n = 36 024) from the version 5 dataset who were not in the original study. Internal validation of original rate models involved fitting a calibration model to compare model parameters between the two samples and bootstrap estimates of bias in the entire version 5 dataset. RESULTS: Remission, continuation and incidence estimates remained stable. Pre-pregnancy prevalence estimates in the validation sample were: anorexia nervosa (AN; 0.1%), bulimia nervosa (BN; 1.0%), binge eating disorder (BED; 3.3%) and eating disorder not otherwise specified-purging disorder (EDNOS-P; 0.1%). In early pregnancy, estimates were: BN (0.2%), BED (4.8%) and EDNOS-P (<0.01%). Incident BN and EDNOS-P during pregnancy were rare. The highest rates were for full or partial remission for BN and EDNOS-P and continuation for BED. CONCLUSIONS: We validated previously estimated rates of remission, continuation and incidence of eating disorders during pregnancy. Eating disorders, especially BED, during pregnancy were relatively common, occurring in nearly one in every 20 women. Pregnancy was a window of remission from BN but a window of vulnerability for BED. Training to detect eating disorders by obstetricians/gynecologists and interventions to enhance pregnancy and neonatal outcomes warrant attention.
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Trastornos de Alimentación y de la Ingestión de Alimentos/diagnóstico , Trastornos de Alimentación y de la Ingestión de Alimentos/epidemiología , Complicaciones del Embarazo/epidemiología , Adulto , Estudios de Cohortes , Diagnóstico Precoz , Trastornos de Alimentación y de la Ingestión de Alimentos/clasificación , Femenino , Humanos , Incidencia , Noruega/epidemiología , Embarazo , Complicaciones del Embarazo/clasificación , Complicaciones del Embarazo/diagnóstico , Sistema de Registros , Remisión EspontáneaRESUMEN
Emerging preclinical and clinical evidence suggests that pregnenolone may be a promising novel therapeutic candidate in schizophrenia. Pregnenolone is a neurosteroid with pleiotropic actions in rodents that include the enhancement of learning and memory, neuritic outgrowth, and myelination. Further, pregnenolone administration results in elevations in downstream neurosteroids such as allopregnanolone, a molecule with neuroprotective effects that also increases neurogenesis, decreases apoptosis and inflammation, modulates the hypothalamic-pituitary-adrenal axis, and markedly increases GABA(A) receptor responses. In addition, pregnenolone administration elevates pregnenolone sulfate, a neurosteroid that positively modulates NMDA receptors. There are thus multiple mechanistic possibilities for pregnenolone as a potential therapeutic agent in schizophrenia, including the amelioration of NMDA receptor hypofunction (via metabolism to pregnenolone sulfate) and the mitigation of GABA dysregulation (via metabolism to allopregnanolone). Additional evidence consistent with a therapeutic role for pregnenolone in schizophrenia includes neurosteroid changes following administration of certain antipsychotics in rodent models. For example, clozapine elevates pregnenolone levels in rat hippocampus, and these increases may potentially contribute to its superior antipsychotic efficacy [Marx et al. (2006a) Pharmacol Biochem Behav 84:598-608]. Further, pregnenolone levels appear to be altered in postmortem brain tissue from patients with schizophrenia compared to control subjects [Marx et al. (2006c) Neuropsychopharmacology 31:1249-1263], suggesting that neurosteroid changes may play a role in the neurobiology of this disorder and/or its treatment. Although clinical trial data utilizing pregnenolone as a therapeutic agent in schizophrenia are currently limited, initial findings are encouraging. Treatment with adjunctive pregnenolone significantly decreased negative symptoms in patients with schizophrenia or schizoaffective disorder in a pilot proof-of-concept randomized controlled trial, and elevations in pregnenolone and allopregnanolone post-treatment with this intervention were correlated with cognitive improvements [Marx et al. (2009) Neuropsychopharmacology 34:1885-1903]. Another pilot randomized controlled trial recently presented at a scientific meeting demonstrated significant improvements in negative symptoms, verbal memory, and attention following treatment with adjunctive pregnenolone, in addition to enduring effects in a small subset of patients receiving pregnenolone longer-term [Savitz (2010) Society of Biological Psychiatry Annual Meeting New Orleans, LA]. A third pilot clinical trial reported significantly decreased positive symptoms and extrapyramidal side effects following adjunctive pregnenolone, in addition to increased attention and working memory performance [Ritsner et al. (2010) J Clin Psychiatry 71:1351-1362]. Future efforts in larger cohorts will be required to investigate pregnenolone as a possible therapeutic candidate in schizophrenia, but early efforts are promising and merit further investigation. This article is part of a Special Issue entitled: Neuroactive Steroids: Focus on Human Brain.
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Antipsicóticos/uso terapéutico , Evaluación Preclínica de Medicamentos , Pregnenolona/uso terapéutico , Ensayos Clínicos Controlados Aleatorios como Asunto , Esquizofrenia/tratamiento farmacológico , Animales , Modelos Animales de Enfermedad , Maleato de Dizocilpina/uso terapéutico , Humanos , Aprendizaje/efectos de los fármacos , Neurotransmisores/metabolismo , Pregnenolona/metabolismo , RatasRESUMEN
BACKGROUND: In mammals, oocyte activation at fertilization is thought to be induced by the sperm-specific phospholipase C zeta (PLCzeta). However, it still remains to be conclusively shown that PLCzeta is the endogenous agent of oocyte activation. Some types of human infertility appear to be caused by failure of the sperm to activate and this may be due to specific defects in PLCzeta. METHODS AND RESULTS: Immunofluorescence studies showed PLCzeta to be localized in the equatorial region of sperm from fertile men, but sperm deficient in oocyte activation exhibited no specific signal in this same region. Immunoblot analysis revealed reduced amounts of PLCzeta in sperm from infertile men, and in some cases, the presence of an abnormally low molecular weight form of PLCzeta. In one non-globozoospermic case, DNA analysis identified a point mutation in the PLCzeta gene that leads to a significant amino acid change in the catalytic region of the protein. Structural modelling suggested that this defect may have important effects upon the structure and function of the PLCzeta protein. cRNA corresponding to mutant PLCzeta failed to induce calcium oscillations when microinjected into mouse oocytes. Injection of infertile human sperm into mouse oocytes failed to activate the oocyte or trigger calcium oscillations. Injection of such infertile sperm followed by two calcium pulses, induced by assisted oocyte activation, activated the oocytes without inducing the typical pattern of calcium oscillations. CONCLUSIONS: Our findings illustrate the importance of PLCzeta during fertilization and suggest that mutant forms of PLCzeta may underlie certain types of human male infertility.
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Infertilidad Masculina/enzimología , Fosfoinositido Fosfolipasa C/metabolismo , Interacciones Espermatozoide-Óvulo/fisiología , Espermatozoides/metabolismo , Sustitución de Aminoácidos , Animales , Sitios de Unión , Calcio/metabolismo , Fertilización/fisiología , Humanos , Immunoblotting , Masculino , Ratones , Modelos Moleculares , Fosfoinositido Fosfolipasa C/química , Fosfoinositido Fosfolipasa C/genética , Mutación Puntual , Estructura Terciaria de ProteínaRESUMEN
Neural cell adhesion molecule (NCAM) is a membrane-bound cell recognition molecule that exerts important functions in normal neurodevelopment including cell migration, neurite outgrowth, axon fasciculation, and synaptic plasticity. Alternative splicing of NCAM mRNA generates three main protein isoforms: NCAM-180, -140, and -120. Ectodomain shedding of NCAM isoforms can produce an extracellular 105-115 kilodalton soluble neural cell adhesion molecule fragment (NCAM-EC) and a smaller intracellular cytoplasmic fragment (NCAM-IC). NCAM also undergoes a unique post-translational modification in brain by the addition of polysialic acid (PSA)-NCAM. Interestingly, both PSA-NCAM and NCAM-EC have been implicated in the pathophysiology of schizophrenia. The developmental expression patterns of the main NCAM isoforms and PSA-NCAM have been described in rodent brain, but no studies have examined NCAM expression across human cortical development. Western blotting was used to quantify NCAM in human postmortem prefrontal cortex in 42 individuals ranging in age from mid-gestation to early adulthood. Each NCAM isoform (NCAM-180, -140, and -120), post-translational modification (PSA-NCAM) and cleavage fragment (NCAM-EC and NCAM-IC) demonstrated developmental regulation in frontal cortex. NCAM-180, -140, and -120, as well as PSA-NCAM, and NCAM-IC all showed strong developmental regulation during fetal and early postnatal ages, consistent with their identified roles in axon growth and plasticity. NCAM-EC demonstrated a more gradual increase from the early postnatal period to reach a plateau by early adolescence, potentially implicating involvement in later developmental processes. In summary, this study implicates the major NCAM isoforms, PSA-NCAM and proteolytically cleaved NCAM in pre- and postnatal development of the human prefrontal cortex. These data provide new insights on human cortical development and also provide a basis for how altered NCAM signaling during specific developmental intervals could affect synaptic connectivity and circuit formation, and thereby contribute to neurodevelopmental disorders.
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Regulación del Desarrollo de la Expresión Génica , Moléculas de Adhesión de Célula Nerviosa/genética , Moléculas de Adhesión de Célula Nerviosa/metabolismo , Corteza Prefrontal/crecimiento & desarrollo , Corteza Prefrontal/metabolismo , Adolescente , Adulto , Envejecimiento/genética , Envejecimiento/metabolismo , Animales , Moléculas de Adhesión Celular Neuronal/genética , Moléculas de Adhesión Celular Neuronal/metabolismo , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Molécula L1 de Adhesión de Célula Nerviosa/genética , Molécula L1 de Adhesión de Célula Nerviosa/metabolismo , Corteza Prefrontal/embriología , Embarazo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estabilidad Proteica , Ratas , Ratas Sprague-Dawley , Ácidos Siálicos/genética , Ácidos Siálicos/metabolismo , Adulto JovenRESUMEN
OBJECTIVE: The current study describes detailed eating behaviors, dieting behaviors, and attitudes about shape and weight in 4023 women ages 25 to 45. METHOD: The survey was delivered on-line and participants were identified using a national quota-sampling procedure. RESULTS: Disordered eating behaviors, extreme weight loss measures, and negative cognitions about shape and weight were widely endorsed by women in this age group and were not limited to White participants. Thirty-one percent of women without a history of anorexia nervosa or binge eating reported having purged to control weight, and 74.5% of women reported that their concerns about shape and weight interfered with their happiness. DISCUSSION: Unhealthy approaches to weight control and negative attitudes about shape and weight are pervasive even among women without eating disorders. The development of effective approaches to address the impact of these unhealthy behaviors and attitudes on the general well-being and functioning of women is required.
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Anorexia Nerviosa/epidemiología , Trastorno por Atracón/epidemiología , Imagen Corporal , Peso Corporal , Bulimia Nerviosa/epidemiología , Adulto , Negro o Afroamericano/estadística & datos numéricos , Anorexia Nerviosa/etnología , Depresores del Apetito/administración & dosificación , Asiático/estadística & datos numéricos , Trastorno por Atracón/etnología , Índice de Masa Corporal , Bulimia Nerviosa/etnología , Restricción Calórica , Diuréticos/administración & dosificación , Ejercicio Físico , Femenino , Hispánicos o Latinos/estadística & datos numéricos , Humanos , Indígenas Norteamericanos/estadística & datos numéricos , Laxativos/administración & dosificación , Persona de Mediana Edad , Prevalencia , Muestreo , Estados Unidos/epidemiología , Vómitos , Población Blanca/estadística & datos numéricosRESUMEN
BACKGROUND AND AIMS: Alpha-gliadin proteins are important for the industrial quality of bread wheat flour, but they also contain many epitopes that can trigger celiac (coeliac) disease (CD). The B-genome-encoded alpha-gliadin genes, however, contain very few epitopes. Controlling alpha-gliadin gene expression in wheat requires knowledge on the processes of expression and deposition of alpha-gliadin protein during wheat grain development. METHODS: A 592-bp fragment of the promotor of a B-genome-encoded alpha-gliadin gene driving the expression of a GUS reporter gene was transformed into wheat. A large number of transgenic lines were used for data collection. GUS staining was used to determine GUS expression during wheat kernel development, and immunogold labelling and tissue printing followed by staining with an alpha-gliadin-specific antibody was used to detect alpha-gliadin protein deposited in developing wheat kernels. The promoter sequence was screened for regulatory motifs and compared to other available alpha-gliadin promoter sequences. KEY RESULTS: GUS expression was detected primarily in the cells of the starchy endosperm, notably in the subaleurone layer but also in the aleurone layer. The alpha-gliadin promoter was active from 11 days after anthesis (DAA) until maturity, with an expression similar to that of a 326-bp low molecular weight (LMW) subunit gene promoter reported previously. An alpha-gliadin-specific antibody detected alpha-gliadin protein in protein bodies in the starchy endosperm and in the subaleurone layer but, in contrast to the promoter activity, no alpha-gliadin was detected in the aleurone cell layer. Sequence comparison showed differences in regulatory elements between the promoters of alpha-gliadin genes originating from different genomes (A and B) of bread wheat both in the region used here and upstream. CONCLUSIONS: The results suggest that additional regulator elements upstream of the promoter region used may specifically repress expression in the aleurone cell layer. Observed differences in expression regulator motifs between the alpha-gliadin genes on the different genomes (A and B) of bread wheat leads to a better understanding how alpha-gliadin expression can be controlled.
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Regulación de la Expresión Génica de las Plantas , Gliadina/genética , Plantas Modificadas Genéticamente/genética , Regiones Promotoras Genéticas , Triticum/genética , Epítopos/genética , Epítopos/metabolismo , Genes Reporteros , Gliadina/metabolismo , Glucuronidasa/genética , Glucuronidasa/metabolismo , Fenotipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Triticum/metabolismoRESUMEN
A dry crust loses its crispness when water migrates into the crust. It is not clear if it is the amount of water absorbed or the water activity ( a w) that leads to a loss of crispness. The hysteresis effect observed when recording a water sorption isotherm allowed us to study the effects of a w and moisture content separately. All experiments were carried out on model bread crusts made from Soissons bread flour. The effect of water content and water activity on the glass transition of model bread crusts was studied in detail using two complimentary techniques: phase transition analysis (PTA) and nuclear magnetic resonance (NMR). The results were compared with sensory data and results from a puncture test, which provided data on acoustic emission and fracture mechanics during breaking of the crusts. The water content of the crust was found to be decisive for the transition point as measured by PTA and NMR. However, both water content and water activity had an effect on perceived crispness and number of force and sound peaks. From this may be concluded that the distribution of the water in the samples with a history of high water content is more inhomogeneous, which results in crispy and less crispy regions, thus making them overall more crispy than samples with the same water content but higher a w.
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Pan/análisis , Sensación , Agua/análisis , Agua/química , Fenómenos Químicos , Química Física , Tecnología de Alimentos , Humanos , Almidón/análisis , Almidón/química , Temperatura de TransiciónRESUMEN
We examined achromatic contrast discrimination in asymptomatic carriers of 11778 Leber's hereditary optic neuropathy (LHON 18 controls) and 18 age-match were also tested. To evaluate magnocellular (MC) and Parvocellular (PC) contrast discrimination, we used a version of Pokorny and Smith's (1997) pulsed/steady-pedestal paradigms (PPP/SPP) thought to be detected via PC and MC pathways, respectively. A luminance pedestal (four 1 degree x 1 degree squares) was presented on a 12 cd/m2 surround. The luminance of one of the squares (trial square, TS) was randomly incremented for either 17 or 133 ms. Observers had to detect the TS, in a forced-choice task, at each duration, for three pedestal levels: 7, 12, 19 cd/m2. In the SPP, the pedestal was fixed, and the TS was modulated. For the PPP, all four pedestal squares pulsed for 17 or 133 ms, and the TS was simultaneously incremented or decremented. We found that contrast discrimination thresholds of LHON carriers were significantly higher than controls' in the condition with the highest luminance of both paradigms, implying impaired contrast processing with no evidence of differential sensitivity losses between the two systems. Carriers' thresholds manifested significantly longer temporal integration than controls in the SPP, consistent with slowed MC responses. The SPP and PPP paradigms can identify contrast and temporal processing deficits in asymptomatic LHON carriers, and thus provide an additional tool for early detection and characterization of the disease.
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Sensibilidad de Contraste , Tamización de Portadores Genéticos , Atrofia Óptica Hereditaria de Leber/genética , Adolescente , Adulto , Discriminación en Psicología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Valores de Referencia , Pruebas de Visión , Agudeza Visual , Vías VisualesRESUMEN
Previous studies of postnatal synaptic development in human frontal cortex have shown that synaptic density rises after birth, reaches a plateau in childhood and then decreases to adult levels by late adolescence. A similar pattern has been seen in nonhuman primate cortex. These earlier studies in human cortex are limited, however, by significant age gaps in study subjects at critical inflection points of the developmental curve. Additionally, it is unclear if synaptic development occurs in different patterns in different cortical layers in prefrontal cortex (PFC). The purpose of this study was to examine synaptic density in human PFC across development by measuring two synaptic marker proteins: synaptophysin (presynaptic), and postsynaptic density protein 95 (PSD-95; postsynaptic). Western blotting was used to assess the relative levels of synaptophysin and PSD-95 in dorsolateral PFC of 42 subjects, distributed in age from 18 weeks gestation to 25 years. In addition, synaptophysin immunoreactivity was examined in each layer of areas 9 and 46 of PFC in 24 subjects, ranging in age from 0.1-25 years. Synaptophysin levels slowly increased from birth until age 5 and then increased more rapidly to peak in late childhood around age 10. Synaptophysin subsequently decreased until the adult level was reached by mid-adolescence, around age 16. PSD-95 levels increased postnatally to reach a stable plateau by early childhood with a slight reduction in late adolescence and early adulthood. The pattern of synaptophysin immunoreactivity seen with immunohistochemistry was similar to the Western experiments but the changes across age were more subtle, with little change by layer within and across age. The developmental patterns exhibited by these synaptic marker proteins expand upon previous studies of developmental synaptic changes in human frontal cortex; synaptic density increases steadily from birth to late childhood, then decreases in early adolescence to reach adult levels by late adolescence.
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Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana/metabolismo , Corteza Prefrontal/crecimiento & desarrollo , Corteza Prefrontal/metabolismo , Sinaptofisina/metabolismo , Adolescente , Adulto , Animales , Western Blotting , Niño , Preescolar , Homólogo 4 de la Proteína Discs Large , Femenino , Edad Gestacional , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Lactante , Recién Nacido , Masculino , Proteínas del Tejido Nervioso/metabolismo , Cambios Post Mortem , Corteza Prefrontal/embriología , Embarazo , Ratas , Ratas Sprague-Dawley , Sinapsis/metabolismoRESUMEN
BACKGROUND AND PURPOSE: The early postnatal period is perhaps the most dynamic phase of white matter development. We hypothesized that the early postnatal development of the corpus callosum and corticospinal tracts could be studied in unsedated healthy neonates by using novel approaches to diffusion tensor imaging (DTI) and quantitative tractography. MATERIALS AND METHODS: Isotropic 2 x 2 x 2 mm(3) DTI and structural images were acquired from 47 healthy neonates. DTI and structural images were coregistered and fractional anisotropy (FA), mean diffusivity (MD), and normalized T1-weighted (T1W) and T2-weighted (T2W) signal intensities were determined in central midline and peripheral cortical regions of the white matter tracts of the genu and splenium of the corpus callosum and the central midbrain and peripheral cortical regions of the corticospinal tracts by using quantitative tractography. RESULTS: We observed that central regions exhibited lower MD, higher FA values, higher T1W intensity, and lower T2W intensity than peripheral cortical regions. As expected, MD decreased, FA increased, and T2W signal intensity decreased with increasing age in the genu and corticospinal tract, whereas there was no significant change in T1W signal intensity. The central midline region of the splenium fiber tract has a unique pattern, with no change in MD, FA, or T2W signal intensity with age, suggesting different growth trajectory compared with the other tracts. FA seems to be more dependent on tract organization, whereas MD seems to be more sensitive to myelination. CONCLUSIONS: Our novel approach may detect small regional differences and age-related changes in the corpus callosum and corticospinal white matter tracts in unsedated healthy neonates and may be used for future studies of pediatric brain disorders that affect developing white matter.
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Cuerpo Calloso/anatomía & histología , Cuerpo Calloso/crecimiento & desarrollo , Imagen por Resonancia Magnética/métodos , Fibras Nerviosas Mielínicas/fisiología , Fibras Nerviosas Mielínicas/ultraestructura , Tractos Piramidales/anatomía & histología , Tractos Piramidales/crecimiento & desarrollo , Femenino , Humanos , Interpretación de Imagen Asistida por Computador/métodos , Recién Nacido , MasculinoRESUMEN
BACKGROUND AND PURPOSE: To determine the sample size needed to provide adequate statistical power in studies of brain volume by MR imaging, we examined the precision and variability of measurements in healthy controls. MATERIALS AND METHODS: A cohort of 52 people (mean age, 25.1 years) was examined at weeks 0 and 12 at 1.5 T. We used an axial multisection T1-weighted sequence and a contiguous proton-attenuation/T2-weighted sequence. Data were registered to a probabilistic brain atlas, and an automated atlas-based program was used to segment brain tissue by type and by lobe. We assumed that there were no changes in volume because there were no intervening neurologic events. Sample sizes required to yield 80% statistical power in detecting a significant difference in volume were calculated for various experimental designs, assuming a patient-control volume difference of 5% or 2%. RESULTS: The precision of most measurements was excellent, but required sample sizes were larger than anticipated. If the goal was to detect a 5% difference in whole brain volume in a 2-sample cross-sectional study, the required sample was 73 patients and 73 controls because brain volume varies between individuals in a way that is not informative about disease effects. For a similar 2-sample longitudinal study, the required sample size was just 5 patients and 5 controls. CONCLUSIONS: Our results argue strongly for longitudinal studies in preference to cross-sectional studies, especially as research budgets decline. Our findings also suggest that there may be more uncertainty than expected in published MR imaging brain volume studies.
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Algoritmos , Encéfalo/anatomía & histología , Interpretación de Imagen Asistida por Computador/métodos , Imagenología Tridimensional/métodos , Imagen por Resonancia Magnética/métodos , Adulto , Encéfalo/fisiología , Femenino , Humanos , Aumento de la Imagen/métodos , Masculino , Tamaño de los Órganos/fisiología , Reproducibilidad de los Resultados , Tamaño de la Muestra , Sensibilidad y EspecificidadRESUMEN
In order to account for the multi-phasic dynamics of photopigment regeneration in human rods, we developed a new model of the retinoid cycle. We first examined the relative roles of the classical and channeling mechanisms of metarhodopsin decay in establishing these dynamics. We showed that neither of these mechanisms alone, nor a linear combination of the two, can adequately account for the dynamics of rhodopsin regeneration at all bleach levels. Our new model adds novel inhibitory interactions in the cycle of regeneration of rhodopsin that are consistent with the 3D structure of rhodopsin. Our analyses show that the dynamics of human rod photopigment regeneration can be accounted for by end-product regulation of the channeling mechanism where all-trans retinal (tral) inhibits the binding of 11-cis retinal to the opsin.tral complex.