Immunohistochemical and molecular detection of equine herpesvirus 1 in Uruguay

Equine herpesvirus 1 (EHV-1) is a major cause of epidemic abortion, neonatal mortality, respiratory disease and neurological disorders in horses. In South America, the virus has been isolated in Brazil, Argentina and Colombia. In Chile pathological findings from one aborted foetus have been reported, and in Uruguay only serological data about EHV-1 activity have been found. Some pathological findings were reported in Uruguay several years ago, but these data have never been officially confirmed. The present work describes the relevant findings of a study of EHV-1 infections in the Uruguayan equine population using polymerase chain reaction (PCR) and histological and immunohistochemical analysis techniques. The sequence analysis of a portion of the glycoprotein C gene amplified by PCR confirmed EHV-1 activity. The real-time PCR revealed the association of the virus with the non-neuropathogenic genotype. This study describes for the first time the immunohistochemical and molecular detection of EHV-1 in Uruguay.Facultad de Ciencias Veterinaria

Revealing the coral habitat effect on benthopelagic fauna diversity in the Santa Maria di Leuca cold-water coral province using different devices and Bayesian hierarchical modelling

Data from two experimental longline surveys and two video inspections carried out in Santa Maria di Leuca cold-water coral province (Mediterranean Sea) during spring-autumn 2010 were used in order to compare the benthopelagic abundance and diversity between coral and non-coral habitats and between different devices. The sampling was carried out in two types of habitat: a coral habitat with carbonate mounds and a non-coral habitat characterized by intermound sea floor. A Bayesian hierarchical modelling approach to accommodate factors influencing community assemblages was used considering the number of species, the Shannon-Wiener diversity index and the two most abundant species represented by the European conger (Conger conger) and blackbelly rosefish (Helicolenus dactylopterus). A relevant effect of the habitat factor was observed for both the number of species and the diversity index, showing a higher species number and diversity index in the coral habitat than in the non-coral habitat. Concerning the relevance of fixed effects from the model on the probability of observing non-zero (positive) abundances, the devices considered, longline and baited lander, did not show different influence for either C. conger or H. dactylopterus. In the case of positive abundance, a relevant device effect was only observed for H. dactylopterus, showing higher abundances for longline than for baited lander. A habitat effect was detected, with positive abundances for both species in the coral habitat. This study proves that structurally complex habitats generated by cold-water corals influence the distribution and diversity of the benthopelagic fauna, and that the use of different devices can provide complementary useful results. Increased knowledge about the role of cold-water corals in the associated benthopelagic fauna could lead to better conservation of one of the most important hot spots of biodiversity in the Mediterranean Sea

Immunohistochemical and molecular detection of equine herpesvirus 1 in Uruguay

Equine herpesvirus 1 (EHV-1) is a major cause of epidemic abortion, neonatal mortality, respiratory disease and neurological disorders in horses. In South America, the virus has been isolated in Brazil, Argentina and Colombia. In Chile pathological findings from one aborted foetus have been reported, and in Uruguay only serological data about EHV-1 activity have been found. Some pathological findings were reported in Uruguay several years ago, but these data have never been officially confirmed. The present work describes the relevant findings of a study of EHV-1 infections in the Uruguayan equine population using polymerase chain reaction (PCR) and histological and immunohistochemical analysis techniques. The sequence analysis of a portion of the glycoprotein C gene amplified by PCR confirmed EHV-1 activity. The real-time PCR revealed the association of the virus with the non-neuropathogenic genotype. This study describes for the first time the immunohistochemical and molecular detection of EHV-1 in Uruguay.Facultad de Ciencias Veterinaria

ADAMTS2 gene dysregulation in T/myeloid mixed phenotype acute leukemia.

Background: Mixed phenotype acute leukemias (MPAL) include acute leukemias with blasts that express antigens of more than one lineage, with no clear evidence of myeloid or lymphoid lineage differentiation. T/myeloid (T/My) MPAL not otherwise specified (NOS) is a rare leukemia that expresses both T and myeloid antigens, accounting for less than 1% of all leukemias but 89% of T/My MPAL. From a molecular point of view, very limited data are available on T/My MPAL NOS. Case presentation: In this report we describe a T/My MPAL NOS case with a complex rearrangement involving chromosomes 5 and 14, resulting in overexpression of the ADAM metallopeptidase with thrombospondin type 1 motif, 2 (ADAMTS2) gene due to its juxtaposition to the T cell receptor delta (TRD) gene segment. Conclusion: Detailed molecular cytogenetic characterization of the complex rearrangement in the reported T/My MPAL case allowed us to observe ADAMTS2 gene overexpression, identifying a molecular marker that may be useful for monitoring minimal residual disease. To our knowledge, this is the first evidence of gene dysregulation due to a chromosomal rearrangement in T/My MPAL NOS. Keywords: Mixed phenotype acute leukemia, ADAMTS2, TRD, Complex chromosomal rearrangement, Promoter swapping, Gene dysregulatio

Contrast-Enhanced Ultrasound Imaging of Prostate Gland in Neutered Dogs

Prostatic neoplasia (PN) occurs in 5–7% of dogs with prostatic disease, with castrated dogs having the same or higher prevalence when compared to intact dogs. Considering the promising results achieved by performing contrast-enhanced ultrasound (CEUS) in intact dogs to detect PN, the present study aimed to acquire data on the prostatic perfusion pattern in neutered dogs. CEUS was performed in 64 neutered dogs, using a 5–7.5 MHz linear transducer with coded harmonic capability, dedicated analytical software, and a second-generation contrast agent, SonoVue. After B-mode evaluation was performed to assess mean prostate volume, the CEUS examination was undertaken. The flow of contrast agent was visible 10 s after injection. The subcapsular vessels were highlighted and produced rapid peripheral rim enhancement. Subsequently, the contrast agent reached the prostatic urethra via the parenchymal arterioles and gradually reached the entire prostate. Perfusion peak intensity (PPI) and time to peak (TTP) values were respectively 45.3% and 34.1 s. The measured parameters were compared with those obtained in previous studies on intact dogs with normal and with pathological patterns. In this study, CEUS showed features that may be promising for its use as a diagnostic tool for early detection of PN in neutered dogs

Differential Effects of Two Isoenergetic Meals Rich in Saturated or Monounsaturated Fat on Endothelial Function in Subjects With Type 2 Diabetes

OBJECTIVE—To examine the acute effects of consumption of monounsaturated (MUFAs) and saturated fatty acids (SAFAs) on endothelial function in subjects with type 2 diabetes

Analysis of CGF biomolecules, structure and cell population: Characterization of the stemness features of CGF cells and osteogenic potential

Concentrated Growth Factors (CGF) represent new autologous (blood-derived biomaterial), attracting growing interest in the field of regenerative medicine. In this study, the chemical, structural, and biological characterization of CGF was carried out. CGF molecular characterization was performed by GC/MS to quantify small metabolites and by ELISA to measure growth factors and matrix metalloproteinases (MMPs) release; structural CGF characterization was carried out by SEM analysis and immunohistochemistry; CGF has been cultured, and its primary cells were isolated for the identification of their surface markers by flow cytometry, Western blot, and real-time PCR; finally, the osteogenic differentiation of CGF primary cells was evaluated through matrix mineralization by alizarin red staining and through mRNA quantification of osteogenic differentiation markers by real-time PCR. We found that CGF has a complex inner structure capable of influencing the release of growth factors, metabolites, and cells. These cells, which could regulate the production and release of the CGF growth factors, show stem features and are able to differentiate into osteoblasts producing a mineralized matrix. These data, taken together, highlight interesting new perspectives for the use of CGF in regenerative medicine

The secretome derived from mesenchymal stromal cells cultured in a xeno-free medium promotes human cartilage recovery in vitro

Osteoarthritis (OA) is a disabling joint disorder causing articular cartilage degeneration. Currently, the treatments are mainly aimed to pain and symptoms relief, rather than disease amelioration. Human bone marrow stromal cells (hBMSCs) have emerged as a promising paracrine mechanism-based tool for OA treatment. Here, we investigate the therapeutic potential of conditioned media (CM) and extracellular vesicles (EVs) isolated from hBMSC and grown in a xeno-free culture system (XFS) compared to the conventional fetal bovine serum-culture system (FBS) in an in vitro model of OA. First, we observed that XFS promoted growth and viability of hBMSCs compared to FBS-containing medium while preserving their typical phenotype. The biological effects of the CM derived from hBMSC cultivated in XFS-and FBS-based medium were tested on IL-1α treated human chondrocytes, to mimic the OA enviroment. Treatment with CM derived from XFS-cultured hBMSC inhibited IL-1α-induced expression of IL-6, IL-8, and COX-2 by hACs compared to FBS-based condition. Furthermore, we observed that hBMSCs grown in XFS produced a higher amount of EVs compared to FBS-culture. The hBMSC-EVs not only inhibit the adverse effects of IL-1α-induced inflammation, but play a significant in vitro chondroprotective effect. In conclusion, the XFS medium was found to be suitable for isolation and expansion of hBMSCs with increased safety profile and intended for ready-to-use clinical therapies