Dynamics of extracellular matrix in ovarian follicles and corpora lutea of mice

Despite the mouse being an important laboratory species, little is known about changes in its extracellular matrix (ECM) during follicle and corpora lutea formation and regression. Follicle development was induced in mice (29 days of age/experimental day 0) by injections of pregnant mare’s serum gonadotrophin on days 0 and 1 and ovulation was induced by injection of human chorionic gonadotrophin on day 2. Ovaries were collected for immunohistochemistry (n=10 per group) on days 0, 2 and 5. Another group was mated and ovaries were examined on day 11 (n=7). Collagen type IV α1 and α2, laminin α1, β1 and γ1 chains, nidogens 1 and 2 and perlecan were present in the follicular basal lamina of all developmental stages. Collagen type XVIII was only found in basal lamina of primordial, primary and some preantral follicles, whereas laminin α2 was only detected in some preantral and antral follicles. The focimatrix, a specialised matrix of the membrana granulosa, contained collagen type IV α1 and α2, laminin α1, β1 and γ1 chains, nidogens 1 and 2, perlecan and collagen type XVIII. In the corpora lutea, staining was restricted to capillary sub-endothelial basal laminas containing collagen type IV α1 and α2, laminin α1, β1 and γ1 chains, nidogens 1 and 2, perlecan and collagen type XVIII. Laminins α4 and α5 were not immunolocalised to any structure in the mouse ovary. The ECM composition of the mouse ovary has similarities to, but also major differences from, other species with respect to nidogens 1 and 2 and perlecan

Association of two newly recognized herpesviruses with interstitial pneumonia in donkeys (Equus asinus)

Over a period of 6 years, antemortem and postmortem examinations were performed on a number of donkeys suffering from respiratory disease. For many cases, initial diagnostic efforts failed to identify an etiology consistent with the pathologic findings. However, retrospective examination of these cases using consensus primer polymerase chain reaction, designed to recognize herpesviruses from all 3 subfamilies of the Herpesviridae, amplified a fragment of the highly conserved herpesvirus DNA polymerase gene from a number of these animals. Two novel herpesviruses, herein designated asinine herpesvirus 4 (AHV4) and asinine herpesvirus 5 (AHV5), were consistently detected in lung tissue from donkeys in which the histopathology was characterized by interstitial pneumonia and marked syncytial cell formation but not in lung tissue from donkeys with evidence of bacterial or verminous pneumonia. Nucleotide sequence and phylogenetic analysis places these new viruses within the Gammaherpesvirinae subfamily and indicates that they are most closely related to the recently identified zebra herpesvirus and wildass herpesvirus as well as equine herpesviruses 2 and 5

Glutamate dehydrogenase as a biomarker for mitotoxicity; insights from furosemide hepatotoxicity in the mouse.

Glutamate dehydrogenase (GLDH) is a liver-specific biomarker of hepatocellular damage currently undergoing qualification as a drug development tool. Since GLDH is located within the mitochondrial matrix, it has been hypothesized that it might also be useful in assessing mitotoxicity as an initiating event during drug-induced liver injury. According to this hypothesis, hepatocyte death that does not involve primary mitochondrial injury would result in release of intact mitochondria into circulation that could be removed by high speed centrifugation and result in lower GLDH activity measured in spun serum vs un-spun serum. A single prior study in mice has provided some support for this hypothesis. We sought to repeat and extend the findings of this study. Accordingly, mice were treated with the known mitochondrial toxicant, acetaminophen (APAP), or with furosemide (FS), a toxicant believed to cause hepatocyte death through mechanisms not involving mitotoxicity as initiating event. We measured GLDH levels in fresh plasma before and after high speed centrifugation to remove intact mitochondria. We found that both APAP and FS treatments caused substantial hepatocellular necrosis that correlated with plasma alanine aminotransferase (ALT) and GLDH elevations. The plasma GLDH activity in both the APAP- and FS- treated mice was not affected by high-speed centrifugation. Interestingly, the ratio of GLDH:ALT was 5-fold lower during FS compared to APAP hepatotoxicity. Electron microscopy confirmed that both APAP- and FS-treatments had resulted in mitochondrial injury. Mitochondria within vesicles were only observed in the FS-treated mice raising the possibility that mitophagy might account for reduced release of GLDH in the FS-treated mice. Although our results show that plasma GLDH is not clinically useful for evaluating mitotoxicity, the GLDH:ALT ratio as a measure of mitophagy needs to be further studied

Modeling Performance and Settlement Windows of Larval Eastern Oyster (\u3ci\u3eCrassostrea virginica\u3c/i\u3e) In Delaware Bay

Oyster population maintenance and growth require a sufficient larval supply competent for metamorphosis and settlement. Larval performance, in terms of growth, development, survival, and metamorphic success, determines the capacity for a larval cohort to effectively settle and establish into an existing population. Exogenous factors influencing larval development include temperature, salinity, food quantity, and food quality. A sufficient diet, composed of balanced protein, lipids, and carbohydrates to meet larval nutritional demands, is required to promote successful metamorphosis. To evaluate the influence of these exogenous factors on oyster settlement potential in Delaware Bay, a well-established biochemically based Crassostrea gigas (Thunberg, 1793) larval model was adapted to simulate Crassostrea virginica (Gmelin, 1791) larval performance under in situ environmental conditions measured during the 2009 to 2011 reproductive seasons at 10 sites across the salinity gradient of Delaware Bay. Variation in the initial egg size and lipid content, and larval food assimilation efficiency was incorporated into the model to represent potential within-cohort phenotypic variability. The middle portion of Delaware Bay along the New Jersey shoreline, bridging the 15-salinity line, generated the most successful larvae each year, whereas the low-salinity reach, on the Delaware side, and Nantuxent Point Reef had more variable success. Survivorship was a function of adequate temperatures and salinities, sufficient food quantity, and favorable food quality defined in part by the protein-to-(lipid-plus-carbohydrate) ratio. Most settlement was predicted by the model to occur between July and September of each year. To validate the model, estimated settlement windows were compared with calculated settlement windows derived from recruitment observations on yearly shell plants. Modeled and recruitment-derived settlement windows agreed well with each other and verified the capacity of the model to accurately forecast in situ larval performance. The oyster larval model, based on measures of lipid, protein, and carbohydrate, successfully passed an important field test, demonstrating the potential of such biochemically based models to reliably evaluate larval performance under real-world conditions