ABCB1 c.-6-180 T > G polymorphism and clinical risk factors in a multi-breed cohort of dogs with refractory idiopathic epilepsy

Epilepsy is the most common chronic neurological disorder in dogs. Approximately 20-30% of dogs do not achieve satisfactory seizure control with two or more anti-epileptic drugs at appropriate dosages. This condition, defined as refractory epilepsy, is a multifactorial condition involving both acquired and genetic factors. The P glycoprotein might play and important role in the pathophysiological mechanism and it is encoded by the ABCB1 gene. An association between a single nucleotide variation of the ABCB1 gene (c.-6-180 T > G) and phenobarbital resistance has previously been reported in a Border collie population with idiopathic epilepsy. To date, the presence and relevance of this polymorphism has not been assessed in other breeds. A multicentre retrospective, case-control study was conducted to investigate associations between ABCB1 c.-6-180 T > G, clinical variables, and refractoriness in a multi-breed population of dogs with refractory idiopathic epilepsy. A secondary aim was to evaluate the possible involvement of the ABCB1 c.-6-180 T > G single nucleotide variation this population. Fifty-two refractory and 50 responsive dogs with idiopathic epilepsy were enrolled. Of these, 45 refractory and 50 responsive (control) dogs were genotyped. The G allele was found in several breeds, but there was no evidence of association with refractoriness (P = 0.69). The uncertain role of the c.-6-180T>G variation was further suggested by an association between the T/T genotype with both refractoriness and responsiveness in different breeds. Furthermore, high seizure density (cluster seizure) was the main clinical risk factor for refractory idiopathic epilepsy (P = 0.003)

Derangement of a Factor Upstream of RARα Triggers the Repression of a Pleiotropic Epigenetic Network

Chromatin adapts and responds to extrinsic and intrinsic cues. We hypothesize that inheritable aberrant chromatin states in cancer and aging are caused by genetic/environmental factors. In previous studies we demonstrated that either genetic mutations, or loss, of retinoic acid receptor alpha (RARalpha), can impair the integration of the retinoic acid (RA) signal at the chromatin of RA-responsive genes downstream of RARalpha, and can lead to aberrant repressive chromatin states marked by epigenetic modifications. In this study we tested whether the mere interference with the availability of RA signal at RARalpha, in cells with an otherwise functional RARalpha, can also induce epigenetic repression at RA-responsive genes downstream of RARalpha.To hamper the availability of RA at RARalpha in untransformed human mammary epithelial cells, we targeted the cellular RA-binding protein 2 (CRABP2), which transports RA from the cytoplasm onto the nuclear RARs. Stable ectopic expression of a CRABP2 mutant unable to enter the nucleus, as well as stable knock down of endogenous CRABP2, led to the coordinated transcriptional repression of a few RA-responsive genes downstream of RARalpha. The chromatin at these genes acquired an exacerbated repressed state, or state "of no return". This aberrant state is unresponsive to RA, and therefore differs from the physiologically repressed, yet "poised" state, which is responsive to RA. Consistent with development of homozygosis for epigenetically repressed loci, a significant proportion of cells with a defective CRABP2-mediated RA transport developed heritable phenotypes indicative of loss of function.Derangement/lack of a critical factor necessary for RARalpha function induces epigenetic repression of a RA-regulated gene network downstream of RARalpha, with major pleiotropic biological outcomes

STUDIO SULLA BIOPSIA TC-GUIDATA DEL POLMONE CON AGO SOTTILE NEL CANE E NEL GATTO

La diagnosi delle patologie polmonari sulla base dell’anamnesi e della visita clinica è spesso difficile. La diagnostica per immagini risulta essere di grande importanza in questo settore. La radiologia è stata considerata per molto tempo la tecnica di elezione per lo studio di queste malattie. Tuttavia spesso non è possibile differenziare una lesione infiammatoria/infettiva da una neoplastica. Risulta quindi necessaria una corretta diagnosi cito-istopatologica per avere una diagnosi accurata, una corretta prognosi ed un preciso piano terapeutico. In medicina umana la TC e la biopsia TC-guidata sono indicate in presenza di lesioni non adeguatamente visualizzate con altre procedure diagnostiche. Nel presente studio sono stati sottoposti a biopsia TC-guidata con ago sottile 30 cani e 9 gatti, di differente sesso, razza e dimensioni. In tutti gli animali sono stati eseguiti esame clinico, esami ematici e le radiografie del torace. Nel presente studio 32 campioni su 39 sono risultati diagnostici. Gli altri 7 casi o a causa di incertezza diagnostica, od in quanto contenevano solo sangue sono stati considerati non diagnostici. Non si sono riscontrate complicazioni gravi, solamente 5 casi di lieve pneumotorace.Diagnosis of pulmonary lesions on the basis of history and physical examination is often challenging. Diagnostic imaging is therefore of paramount importance in this field. Radiology has traditionally been considered the elective diagnostic procedure for these diseases. Nonetheless it is often not possible to differentiate inflammatory/infectious lesions from neoplastic disease. A correct cyto-histopathologic diagnosis is therefore needed for an accurate diagnosis and subsequent prognostic and therapeutic plan. In human medicine TC and TC-guided biopsy are indicated in the presence of lesions which are not adequately imaged with other diagnostic procedures. In the present study 30 dogs and 9 cats of different sex, breed and size underwent TCguided lung fine-needle aspiration. Clinical examination, haematology and chest radiography were performed on all animals. In this study 32 samples out of 39 were diagnostic. Other 7 cases either because of uncertainty or only blood was aspirated, were considered non diagnostic. Only mild pneumothorax was seen in 5 cases. No major complications were encountered

Spinal cord neurenteric cyst: clinical and diagnostic findings and long term follow-up in two dogs

Non present

Effect of cilostamide treatment on the intercellular coupling, meiotic and embryonic developmental competence in horse oocyte-cumulus cells complexes of different origins

Design of new strategies to improve in vitro embryo production (IVP) efficiency, such as pre-maturation culture with cilostamide (CILO PMC) aimed to prolong oocytes-cumulus cells gap junction-mediated communications (GJC) functionality while delaying meiotic resumption, should consider the high heterogeneity of cumulus-oocyte complexes (COCs) retrieved from ovarian follicles. In the mare, main factors contributing to this heterogeneity are: the follicle diameter, the cumulus oophorus morphology and the reproductive seasonality. Thus, the objectives of this study were: 1) to identify a population of equine oocytes that would benefit from a CILO PMC, based on the assessment the GJC functionality by Lucifer Yellow microinjection, the oocyte chromatin configuration by DNA staining, and the meiotic competence after in vitro maturation (IVM), in COCs of different origins, since these parameters are indicative of the oocyte metabolic state and 2) to assess the effect of CILO PMC, in COCs of different origins, on the GJC functionality, the meiotic and embryonic developmental competence after IVM and intracytoplasmic sperm injection (ICSI). COCs with Compact (Cp) or Expanded (Ex) cumulus oophorus were collected from 2 cm follicles during winter, spring-summer and fall, and cultured with our standard IVM protocol (CTRL) or prematurated in the same medium with 10-4 UI r-hFSH/ml and 10 \u3bcM Cilostamide (CILO PMC) for different times. When meiotic and developmental were assessed, COCs were subjected to CILO PMC for 6 h and than to standard IVM and ICSI. Data were analyzed by Chi square test. Overall, our results suggest that COCs that would more probably benefit from CILO PMC are those with Cp cumuli from <1 and 1-2 cm follicles collected during spring-summer and fall. In fact, they have a lower meiotic competence after 24h IVM (P<0.05), with a mean MII rate of 40%, compared to all the other groups, in which MII rate is around 60%. Moreover, they show a higher % (P<0.05) of COC with open GJC (70%), when compared with both Cp COC in the winter season and Exp COC from all follicular classes in all seasons tested, in which more that 50% of COCs have closed GJC. Independently from the season, chromatin configuration analysis reveals that oocytes with Cp cumuli from 1-2 cm follicles are in a more advanced stage of differentiation since their chromatin is more condensed (P<0.05). Study on GJC indicates that functionality can be maintained up to 10h in Cp COCs from 1-2 cm using CILO PMC when compared to the CTRL group, in which a major GJC drop is detected at 10h. In fact, COCs with open GJC are 74% and 44% in CILO PMC and CTRL groups respectively at 10 h (P<0.05) On the contrary, the exact timing for the treatment of Cp COCs from <1 cm follicles remains to be determined since GJC functionality drops after 16h in the CTRL group, with only 14% of COCs with open GJC, and no differences are observed between groups. Finally, initial evaluation of the effect of CILO PMC for 6h indicates that it does not affect the rate of embryos obtained after ICSI. Evaluation of embryo quality is in progress to assess whether CILO PMC could improve IVP efficiency beyond embryo yield. In conclusion, our data could be useful in setting up IVM strategies to improve horse IVP efficiency as well as the understanding of horse oocyte biology. Funding: Grant n. 26096200 "Ex Ovo Omnia" Regione Sardegna-Lombardia and \u2018Dote Ricerca\u2019 FSE, Regione Lombardia