RESUMEN
Expression of immune function genes within follicle cells has been reported in ovaries from many species. Recent work from our laboratory showed a direct effect of the monocyte chemoattractant protein 1/C-C motif chemokine receptor 2 system within the feline cumulus oocyte complex, by increasing the mRNA levels of key genes involved in the ovulatory cascade in vitro. Studies were designed to evaluate if C-C motif chemokine receptor 2 acts as a novel mediator of the ovulatory cascade in vitro. Therefore, feline cumulus oocyte complexes were cultured in the presence or absence of a highly selective C-C motif chemokine receptor 2 antagonist together with known inducers of cumulus-oocyte expansion and/or oocyte maturation to assess mRNA expression of key genes related to periovulatory events in other species as well as oocyte maturation. Also, the effects of recombinant monocyte chemoattractant protein 1 on spontaneous or gonadotrophin-induced oocyte maturation were assessed. This is an in vitro system using isolated cumulus oocyte complexes from feline ovaries. The present study reveals the modulation of several key ovulatory genes by a highly selective C-C motif chemokine receptor 2 antagonist. However, this antagonist was not enough to block the oocyte maturation induced by gonadotropins or amphiregulin. Nonetheless, recombinant monocyte chemoattractant protein 1 had a significant effect on spontaneous oocyte maturation, increasing the percentage of metaphase II stage oocytes in comparison to the control. This is the first study in any species to establish C-C motif chemokine receptor 2 as a mediator of some actions of the mid-cycle gonadotrophin surge.
Asunto(s)
Ovulación/genética , Receptores CCR2/fisiología , Animales , Gatos , Células Cultivadas , Células del Cúmulo/metabolismo , Células del Cúmulo/fisiología , Femenino , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oocitos/metabolismo , Oocitos/fisiología , Oogénesis/genética , Folículo Ovárico/metabolismo , Folículo Ovárico/fisiologíaRESUMEN
Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.
Asunto(s)
Leucosis Bovina Enzoótica/diagnóstico , Virus de la Leucemia Bovina/fisiología , Provirus/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Carga Viral/métodos , Animales , Bovinos , Pruebas Diagnósticas de Rutina/normas , Laboratorios/normas , Virus de la Leucemia Bovina/genética , ARN Viral/genética , Carga Viral/normasRESUMEN
We present a numerical method to compute the acoustic field scattered by finite perforated elastic plates. A boundary element method is developed to solve the Helmholtz equation subjected to boundary conditions related to the plate vibration. These boundary conditions are recast in terms of the vibration modes of the plate and its porosity, which enables a direct solution procedure. A parametric study is performed for a two-dimensional problem whereby a cantilevered perforated elastic plate scatters sound from a point quadrupole near the free edge. Both elasticity and porosity tend to diminish the scattered sound, in agreement with previous work considering semi-infinite plates. Finite elastic plates are shown to reduce acoustic scattering when excited at high Helmholtz numbers k0 based on the plate length. However, at low k0, finite elastic plates produce only modest reductions or, in cases related to structural resonance, an increase to the scattered sound level relative to the rigid case. Porosity, on the other hand, is shown to be more effective in reducing the radiated sound for low k0. The combined beneficial effects of elasticity and porosity are shown to be effective in reducing the scattered sound for a broader range of k0 for perforated elastic plates.
RESUMEN
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed livestock which has a drastic economic impact for affected countries. Although FMDV is distributed worldwide, many regional programs have been effective eradicating this agent. In Argentina, as in many other regions of South America, the combination of a systematic vaccination plan, together with an effective detection system capable of differentiating infection from vaccination, has been successful for eradicating this agent from the country. The properties of recombinant 3AB1 FMDV non-structural protein (r3AB1 FMDV-NSP), as a marker for the detection of antibodies to differentiate between cattle infected and vaccinated with FMDV, have been described previously. The goal of the present study was to validate the 3AB1 ELISA using a well characterized serum panel from Argentina (n=559) including eight national and one international reference sera. Overall, the 3AB1 ELISA demonstrated good feasibility, repeatability, reproducibility, analytical sensitivity and specificity, and accuracy. The results from the 3AB1 ELISA when compared with those obtained from the OIE index test (NCPanaftosa screening) showed a similar performance of both tests [diagnostic sensitivity=84% (C.I.=79-88%) and 80% (C.I.=75-85%), respectively; and diagnostic specificity=98.6% (C.I.=97-100%) and 95% (C.I.=91-98%), respectively]. The present work proposes the 3AB1 ELISA as an alternative to imported kits for FMD internal screening and transboundary sero-surveillance.