Plasma endothelin-1 levels rise in diabetes and after exposure to contrast media suggesting a role in progressive diabetic and acute radiocontrast nephropathies. Here we studied individual and ...combined effects of streptozotocin-induced diabetes and contrast media on renal endothelin converting enzyme-1 levels in the rat. In vivo, medullary (but not cortical) endothelin converting enzyme protein gradually increased 4 to 5-fold following the induction of diabetes or after the administration of contrast media but rose 15-fold when diabetic rats were given contrast media. Changes in mRNA expression paralleled those of the protein. Immunohistochemistry confirmed that increased tubular and endothelial cell endothelin converting enzyme-1 were most pronounced in the medulla. In vitro, endothelin-1 levels increased 3-fold following incubation of endothelial cells with media high in glucose or with contrast and 4-fold with their combination. Endothelin converting enzyme-1 protein and mRNA expression changed in a similar pattern while prepro endothelin-1 mRNA increased with each insult but not in an additive way. Our study shows that diabetes and contrast media up-regulate renal medullary endothelin converting enzyme-1 expression and synthesis.
Introduction
We assessed sex differences in behavioural and neural responses to rectal pain stimuli in healthy subjects.
Methods
In age‐ and body mass index‐matched healthy subjects (n = 15 men, 15 ...women), rectal sensory and pain thresholds were assessed with a pressure‐controlled barostat device. The blood oxygen level‐dependent response during cued anticipation and painful stimulation was measured using functional magnetic resonance imaging (fMRI). Retrospective pain evaluations were accomplished with visual analogue scales. For fMRI data, region‐of‐interest (ROI) analyses and additional whole‐brain analyses were carried out.
Results
There were no sex differences in rectal thresholds or pain ratings. ROI analyses revealed comparable distension‐induced activation of the thalamus, somatosensory cortex, insula and dorsolateral prefrontal cortex (DLPFC). Only in additional whole‐brain analyses did we find increased activation in women in DLPFC and middle temporal gyrus during pain anticipation and in the cerebellum and medial frontal gyrus during pain. A significant inverse association between rectal pain threshold and distension‐induced activation in virtually all ROIs was found in women. In men, pain thresholds and insula activation were positively correlated, as were pain ratings and anterior cingulate cortex activation.
Conclusions
Healthy men and women do not differ in behavioural measures of visceral pain sensitivity. The pattern of neural activation is comparable in the majority of pain‐processing brain regions, although women may differ in the activation of DLPFC which could reflect sex differences in cognitive–emotional pain regulation. Women with lower pain thresholds showed greater neural responses, which may be relevant in the pathophysiology of visceral hyperalgesia.
Newcastle disease (ND) is prevalent worldwide and causes significant clinical and economic losses to the poultry industry. Current vaccine programs using live attenuated vaccines and inactivated ...vaccines have limitations, and new vaccines with distinct features are needed. To offer an alternative solution to control ND, a turkey herpesvirus vector Newcastle disease vaccine (HVT/ND) expressing the fusion gene of Newcastle disease virus (NDV) has been developed. First, immunogenicity of the HVT/ND was evaluated in specific-pathogen-free layer chickens after vaccination by the in ovo route to 18-day-old embryos or by the subcutaneous route to 1-day-old chicks. Antibodies against NDV were detected at 24 days of age using a commercial NDV enzyme-linked immunosorbent assay (ELISA) kit and the hemagglutination inhibition test. At least 90% of chickens were protected against challenge with velogenic neurotropic NDV Texas GB strain (genotype II; pathotype velogenic) at 4 wk of age, while none of the nonvaccinated, challenged controls were protected from challenge. Second, the age at which a vaccinated chicken elicits an immunologic response to the HVT/ND prepared for this study, and thus is protected from ND virus, was assessed in commercial broiler chickens after in ovo vaccination of 18-day-old embryos. Challenge was conducted using a low-virulence NDV strain (genotype II; pathotype lentogenic) via the respiratory tract each week between 1 and 5 wk of age, in order to mimic the situation in areas where virulent NDV strains do not normally exist and low-virulence strains cause mild respiratory symptoms leading to economic losses. Protection was evaluated by the presence or absence of isolated virus from tracheal swabs at 5 days postchallenge. Partial protection was observed at 3 wk of age, when 6 out of 10 (60%) chickens were protected. Full protection was obtained at 4 and 5 wk of age, when 9 out of 10 (90%) and 10 out of 10 (100%) chickens were protected, respectively. Finally, protection against challenge with virulent Texas GB strain at 19 wk of age was evaluated in commercial female layer chickens vaccinated at 1 day of age with HVT/ND. All of the vaccinated chickens were protected, while all of the challenge controls succumbed to the challenge. Furthermore, anti-NDV antibodies measured by ELISA were maintained through 50 wk of age. Protección y respuesta de anticuerpos por una vacuna contra la enfermedad de Newcastle con un herpesvirus de los pavos como vector. La enfermedad de Newcastle (ND) es prevalente en todo el mundo y causa pérdidas clínicas y económicas significativas para la industria avícola. Los programas de vacunación actuales con vacunas vivas atenuadas y vacunas inactivadas tienen limitaciones, y se necesitan nuevas vacunas con características diferentes. Para ofrecer una solución alternativa para controlar a la enfermedad de Newcastle, se desarrolló una vacuna contra la enfermedad de Newcastle utilizando a un herpesvirus de pavo como vector (HVT/ND) que expresa el gene de fusión del virus de la enfermedad de Newcastle (NDV). En primer lugar, se evaluó la inmunogenicidad de la vacuna HVT/ND en aves de postura libres de patógenos específicos después de la vacunación in ovo a los 18 días de desarrollo embrionario, o por vía subcutánea en pollos de un día de edad. Se detectaron anticuerpos contra la enfermedad de Newcastle a los 24 días de edad con un estuche comercial del ensayo de inmunoabsorción con enzimas ligadas (ELISA) para Newcastle y por la prueba de inhibición de la hemaglutinación. Al menos el 90% de los pollos estuvieron protegidos contra el desafío con la cepa velogénica neurotrópica GB Texas (genotipo II; patotipo velogénico) a las cuatro semanas de edad, mientras que ninguna de las aves control no vacunadas, estuvo protegida ante el desafío. En segundo lugar, se evaluó la edad a la que los pollos vacunados desarrollaban una respuesta inmunológica contra la vacuna HVT/ND para este estudio y por lo tanto la protección contra el virus de Newcastle fue evaluada en pollos de engorde comerciales después de la vacunación in ovo a los 18 días de desarrollo embrionario. Se llevó a cabo un desafío usando una cepa de baja virulencia (genotipo II; patotipo lentógénico) a través de las vías respiratorias cada semana entre las semanas una y cinco de edad, con el fin de imitar la situación en áreas en las que normalmente no existen cepas virulentas de Newcastle y las cepas de baja virulencia causan síntomas respiratorios leves que conducen a pérdidas económicas. La protección se evaluó por la presencia o ausencia de aislamiento viral a partir de hisopos traqueales a los cinco días después del desafío. Se observó protección parcial a las tres semanas de edad, cuando se protegieron seis de diez (60%) pollos. Se obtuvo protección total a las cuatro y cinco semanas de edad, cuando nueve de diez (90%) y diez de diez (100%) pollos estuvieron protegidos, respectivamente. Por último, se evaluó la protección contra el desafío con la cepa virulenta GB Texas a las 19 semanas de edad en gallinas de postura comerciales vacunadas al día de edad con la vacuna HVT/ND. Todas las aves vacunadas estaban protegidas, mientras que todos los controles de desafío murieron ante la exposición. Además, los anticuerpos contra Newcastle determinados por ELISA se mantuvieron hasta las 50 semanas de edad.
The aim of the study was to analyse effects of psychological stress on the neural processing of visceral stimuli in healthy women. The brain functional magnetic resonance imaging blood oxygen ...level‐dependent response to non‐painful and painful rectal distensions was recorded from 14 healthy women during acute psychological stress and a control condition. Acute stress was induced with a modified public speaking stress paradigm. State anxiety was assessed with the State‐Trait‐Anxiety Inventory; chronic stress was measured with the Perceived Stress Questionnaire. During non‐painful distensions, activation was observed in the right posterior insular cortex (IC) and right S1. Painful stimuli revealed activation of the bilateral anterior IC, right S1, and right pregenual anterior cingulate cortex. Chronic stress score was correlated with activation of the bilateral amygdala, right posterior IC (post‐IC), left periaqueductal grey (PAG), and right dorsal posterior cingulate gyrus (dPCC) during non‐painful stimulation, and with activation of the right post‐IC, right PAG, left thalamus (THA), and right dPCC during painful distensions. During acute stress, state anxiety was significantly higher and the acute stress – control contrast revealed activation of the right dPCC, left THA and right S1 during painful stimulation. This is the first study to demonstrate effects of acute stress on cerebral activation patterns during visceral pain in healthy women. Together with our finding that chronic stress was correlated wit the neural response to visceral stimuli, these results provide a framework for further studies addressing the role of chronic stress and emotional disturbances in the pathophysiology of visceral hyperalgesia.
Adaptation to hypoxic environment is conferred through hypoxia-inducible transcription factors (HIFs). We have previously shown that the HIF system is transiently activated in vivo in ...radiocontrast-induced acute renal failure, associated with profound hypoxia in the renal medulla. Medullary thick ascending limbs (mTALs), the most affected nephron segments in this model, were virtually unable to mount an adaptive HIF response. Here, we study correlations between oxygenation, HIF activation, and cell viability in a related ex vivo model, the isolated perfused rat kidney (IPK). In IPKs perfused with cell-free oxygenated medium, severe medullary hypoxic damage developed, affecting 42±9% of mTALs in the mid-inner stripe. HIF-1α tubular immunostaining was noted with a zonal and tubular pattern largely similar to our findings in vivo: in 34±3% of collecting ducts (CDs) within the mid-inner stripe and extensively in the papillary tip, whereas mTALs were all HIF-negative. In IPKs supplemented with RBCs (improved oxygen supply), mTAL damage was totally prevented and CDs’ HIF expression was attenuated (22±4%). By contrast, although measures designed to reduce medullary hypoxia by decreasing tubular reabsorptive activity (furosemide, ouabain, or high-albumin-non-filtering system) reduced mTAL damage, all paradoxically resulted in increased HIF expression in CDs (51±4%), and 17±3% of mTALs became immunostained as well. Our data confirm that CDs and mTALs have markedly different HIF responses, which correlate with their viability under hypoxic stress. mTALs transcriptional adaptation occurs within a narrow hypoxic range, and it appears that workload reduction can shift mTALs into this window of opportunity for HIF activation and survival.
In this paper, we put forward a novel fusion framework that mixes together label fields instead of observation data as is usually the case. Our framework takes as input two label fields: a quickly ...estimated and to-be-refined segmentation map and a spatial region map that exhibits the shape of the main objects of the scene. These two label fields are fused together with a global energy function that is minimized with a deterministic iterative conditional mode algorithm. As explained in the paper, the energy function may implement a pure fusion strategy or a fusion-reaction function. In the latter case, a data-related term is used to make the optimization problem well posed. We believe that the conceptual simplicity, the small number of parameters, the use of a simple and fast deterministic optimizer that admits a natural implementation on a parallel architecture are among the main advantages of our approach. Our fusion framework is adapted to various computer vision applications among which are motion segmentation, motion estimation and occlusion detection.
Postsurgical outcome measures are crucial to define the efficacy of perioperative pain management; however, it is unclear which are most appropriate. We conducted a prospective study aiming to assess ...sensitivity-to-change of patient-reported outcome measures assessing the core outcome set of domains pain intensity (at rest/during activity), physical function, adverse events, and self-efficacy.
Patient-reported outcome measures were assessed preoperatively, on day 1 (d1), d3, and d7 after four surgical procedures (total knee replacement, breast surgery, endometriosis-related surgery, and sternotomy). Primary outcomes were sensitivity-to-change of patient-reported outcome measures analysed by correlating their changes (d1-d3) with patients' global impression of change and patients' specific impression of change items as anchor criteria. Secondary outcomes included identification of baseline and patient characteristic variables explaining variance in change for each of the scales and descriptive analysis of various patient-reported outcome measures from different domains and after different surgeries.
Of 3322 patients included (18 hospitals, 10 countries), data from 2661 patients were analysed. All patient-reported outcome measures improved on average over time; the median calculated sensitivity-to-change for all patient-reported outcome measures (overall surgeries) was 0.22 (range: 0.07-0.31, scale: 0-10); all changes were independent of baseline data or patient characteristics and similar between different procedures.
Pain-related patient-reported outcome measures have low to moderate sensitivity-to-change; those showing higher sensitivity-to-change from the same domain should be considered for inclusion in a core outcome set of patient-reported outcome measures to assess the effectiveness and efficacy of perioperative pain management.
The virulence of low pathogenicity (LP) type A H7N2 avian influenza virus (AIV) isolates recovered from chickens in Delaware and the eastern shore of Maryland in 2004 was evaluated. Three-week-old ...leghorn- and broiler-type chickens and turkeys were inoculated via the conjunctival sac with 103.5–104.0 50% embryo infectious dose (EID50) of virus per bird with A/chicken/Delaware/Viva/04, A/chicken/Delaware/Hobo/04, and A/chicken/Maryland/Minh Ma/04. In broilers, the viruses produced respiratory signs, airsacculitis, and microscopic lesions in the trachea and lung. In contrast, signs and lesions were less severe in turkeys, and they were rarely observed in specific-pathogen-free (SPF) leghorns. In broilers and SPF leghorns, AIV peaked on day 3 postinoculation (PI), based on virus isolation and real-time reverse transcription-polymerase chain reaction, and antigen capture testing. Infection in turkeys peaked on day 7 PI. Serum antibodies generally were detected earlier in broilers (day 7 PI) than in turkeys or SPF leghorns (day 14 PI) using agar gel immunodiffusion, hemagglutination-inhibition, and the enzyme-linked immunosorbent assay. A second trial was performed to further examine the disease susceptibility of the leghorn chicken given the comparatively mild responses noted in the first trial. A 10-fold higher dose of 104.5–105.0EID50 per chick given via the conjunctival sac was used. In addition, commercial-type leghorns were tested as were chicks from the SPF leghorn source. The higher AIV dose resulted in more rapid and consistent rates of infection and higher serum antibody responses in both types of leghorn chickens. However, as observed in the first trial, clinical signs and microscopic lesions in both types of leghorns were infrequent and very mild. These findings indicate leghorn-type chickens, which are commonly used for pathogenicity assessments because of their availability, may not be the most suitable host for evaluating the virulence potential of LP AIV. Abbreviations: AGID = agar gel immunodiffusion; AI = avian influenza; AIV = avian influenza virus; CAS = chorioallantoic sac; Ct = cycle threshold; EID50 = embryo infectious dose50; ELD50 = embryo lethal dose50; ELISA = enzyme-linked immunosorbent assay; HA = hemagglutination; HI = hemagglutination-inhibition; HP = high pathogenicity; LP = low pathogenicity; NVSL = National Veterinary Service Laboratory; PBS = phosphate-buffered saline; PI = postinoculation; RRT-PCR = real-time reverse transcription-polymerase chain reaction; SPF = specific-pathogen-free Virulencia en pollos de engorde, aves leghorn y pavos, de aislados de virus H7N2 de influenza aviar de baja patogenicidad provenientes de la península de Delmarva. Se evaluó la virulencia de aislamientos del virus de influenza aviar tipo A H7N2 aislados en el año 2004 a partir de aves domésticas en los estados de Delaware y la costa este de Maryland. Aves de la línea de postura (leghorn), pollos de engorde y pavos de tres semanas de edad fueron inoculados por vía ocular con una dosis infecciosa 50% para embrión de pollo de 103.5–104.0 de los siguientes virus: A/Pollo/Delaware/Viva/04, A/Pollo/Delaware/Hobo/04 y A/Pollo/Maryland/Minh Ma/04. En los pollos de engorde los virus produjeron signos respiratorios, aerosaculitis y lesiones microscópicas en la tráquea y pulmón. En contraste, las lesiones fueron menos severas en los pavos y fueron escasamente observadas en las aves leghorn libres de patógenos específicos. Mediante aislamiento viral, la prueba de reacción en cadena por la polimerasa transcriptasa reversa en tiempo real y pruebas de captura de antígenos, se determinó que en los pollos de engorde y en las aves libres de patógenos específicos los virus de influenza aviar alcanzaron un título máximo al tercer día post inoculación. La infección en los pavos alcanzó un título máximo siete días posteriores a la infección. Utilizando la prueba de inmunodifusión en agar, la prueba de inhibición de la hemoaglutinación y la prueba de inmunoensayo asociado a enzimas, los anticuerpos séricos se detectaron más temprano en pollos de engorde (día siete post inoculación) que en los pavos o las aves libres de patógenos (14 días de edad). Se realizó un segundo experimento con la finalidad de examinar más a profundidad la susceptibilidad de las aves leghorn a los virus de influenza aviar debido a las reacciones comparativamente leves observadas en el primer experimento. Para el segundo experimento se administró en la conjuntiva una dosis 10 veces más alta (104.5–105.0dosis infecciosa 50% para embrión de pollo) por ave. Adicionalmente, se evaluaron aves tipo leghorn comerciales, así como las aves leghorn libres de patógenos específicos. La dosis mas alta del virus de influenza aviar resultó en tasas de infección más rápidas y sostenidas, así como en mayores títulos de anticuerpos séricos en ambos tipos de aves leghorn. Sin embargo, como se observó en el primer experimento, los signos clínicos y las lesiones microscópicas fueron poco comunes y muy leves en ambos tipos de aves leghorn. Estos hallazgos indican que las aves tipo leghorn, que son comúnmente utilizadas debido a su disponibilidad para las evaluaciones de patogenicidad, tal vez no sean el huésped mas adecuado para evaluar el potencial de virulencia de los virus de influenza aviar de baja patogenicidad.
Aim
Von Hippel‐Lindau protein (VHL) provides the degradation of hypoxia‐inducible factor (HIF). Tetracycline‐induced, Pax8‐rtTA‐based knockout of VHL (VHL‐KO) affects all renal tubules and periportal ...hepatocytes and leads to sustained upregulation of HIF. Here, we study the phenotype of VHL‐KO in both organs, the time course of changes, and long‐term morpho‐functional outcome.
Methods
Mice with doxycycline‐induced VHL‐KO and controls (CON) were followed for up to 9 months. Systemic and tissue parameters were evaluated using clinical chemistry, histology, immunohistochemistry, RT‐PCR and in situ hybridisation.
Results
At day 3 following VHL‐KO, substantial abundance of HIF‐1α and ‐2α was detected in the nuclei of hepatocytes and renal tubular epithelia. Hypoxia, induced by bleeding anaemia, did not further augment HIF signal. Erythropoietin mRNA was detectable in hepatocytes but not in the kidney. Vascular endothelial growth factor mRNA was upregulated in kidney but not in liver. At day 7 following VHL‐KO, the renal capillary density was enhanced, reaching its maximum at day 14. Blood haemoglobin increased constantly up to day 28 (23.3 vs. 15.8 g dL−1, VHL‐KO vs. CON). Thereafter, it was kept within the normal range by weekly blood collections. Pathological changes were absent from kidney and liver 9 months after VHL‐KO.
Conclusions
Inducible, Pax8‐rtTA‐based deletion of VHL leads to organ‐specific expression of epithelial HIF and erythropoietin in liver and kidney without causing pathological changes. Uniform, maximal and sustained HIF activation along the renal tubule may serve to study the potential benefits of hypoxia adaptation in experimental renal injury.
Background. Indirect evidence suggests that hypoxia contributes to the pathophysiology of rhabdomyolysis-induced acute kidney injury (AKI). However, the cellular location and kinetics of hypoxia, as ...well as potential hypoxia adaptation are unclear. Methods. Rhabdomyolysis was induced in rats by IM glycerol (GLY) injection, which largely recapitulates the full clinical syndrome. Additional rats received IV myoglobin (MYO), in order to assess the contribution of MYO per se. We performed immunohistochemistry for hypoxia markers pimonidazole (PIM) adducts and hypoxia-inducible factors (HIFs) and the cell-protective HIF target gene heme oxygenase-1 (HO-1). Furthermore, we sought a potential negative feedback loop to terminate HIF activation, driven by HIF prolyl-hydroxylase-2 (PHD-2). Results. In GLY, progressive tubular injury, mainly of proximal tubules (PT), developed over time, but its extent was heterogeneous. PIM, HIFα and HO-1 were all absent in controls, but strongly positive in GLY, with a specific spatio-temporal pattern. In PT, (a) PIM was detectable throughout the study with a maximum at 6 h, (b) HIF was activated only at 3 h and (c) HO-1 and PHD-2 appeared at 6 h and persisted at a lower level at 24 h. Apart from tubular cast formation, MYO did not cause overt tissue damage, but led to strong activation of HIFs, in a pattern similar to 3 h of GLY. Conclusions. Our data suggest that renal hypoxia occurs in rhabdomyolysis, and that MYO, at least partly, contributes to hypoxia generation. Since in the most affected tubules transcriptional hypoxia adaptation is transient and inhomogeneous, pharmacologic HIF enhancement holds the potential to improve outcome in rhabdomyolysis-induced AKI.
PDF
