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)

Exercise-induced bronchospasm in children

Neste trabalho, são revisados conceitos atuais sobre definição, fisiopatologia, diagnóstico clínico e funcional pulmonar e tratamento do broncoespasmo induzido pelo exercício em crianças. O broncoespasmo induzido pelo exercício é o estreitamento da via aérea desencadeado por atividade física em pessoas com reatividade brônquica aumentada. Caracteriza-se por vários graus de obstrução ao fluxo aéreo, ocorrendo minutos após a finalização do exercício. Pode seguir-se também uma resposta tardia, especialmente em crianças. Atinge 15% da população geral, mais freqüentemente asmáticos. A apresentação clínica é variável. O diagnóstico é confirmado por uma queda no pico de fluxo expiratório (PFE) ou no volume expiratório final no primeiro segundo (VEF1) de 15% ou mais após o exercício, ou uma redução no fluxo expiratório forçado de 25 a 75% da capacidade vital (FEF25-75) de 20%. Várias teorias tentam explicar sua fisiopatologia. Os beta2-agonistas são, até o momento, as drogas mais usadas no tratamento e profilaxia do broncoespasmo induzido pelo exercício. Inibidores da degranulação dos mastócitos e antagonistas dos leucotrienos também são utilizados. O broncoespasmo induzido pelo exercício deve ser especialmente lembrado naqueles pacientes asmáticos com sintomas ao exercício. Embora muito já se conheça em termos de alternativas terapêuticas, esta é uma área ainda parcialmente explorada.We review the concept, pathophysiology, clinical diagnosis, pulmonary function tests, and treatment of exercise-induced bronchospasm in children. Exercise-induced bronchospasm is the acute narrowing of the airway that is triggered by vigorous physical activity in individuals with airway hyperreactivity. It is characterized by several degrees of obstruction occurring some minutes after the end of exercise. A late response may also occur, usually in children. Exercise-induced bronchospasm in children affects up to 15% of general population, and it is more prevalent in asthmatics. Diagnosis is suggested by typical history, and confirmed by specific tests. The clinical presentation is variable. Its diagnosis is confirmed by a recorded fall in the peak expiratory flow (PEF) or forced expiratory volume in 1 second (FEV1) of 15% or higher after exercise, or by a decrease in the forced expiratory flow at 25-75% (FEF25-75) of 20% in relation to baseline measurements. Many theories try to explain the pathophysiology of this condition. Beta-2 agonists are the most common drugs used in the treatment of exerciseinduced bronchospasm and in prophylaxis. Mast cell stabilizing agents and leukotriene antagonists can also be used. Exercise-induced bronchospasm should be remembered in asthmatic patients that present symptoms on exercising. Although there is a large amount of information available about therapeutic options, there is still much to be explored in this field of study

Infraestrutura e adesão à higienização das mãos: desafios à segurança do paciente

Considerando a importância das mãos na cadeia de transmissão de microrganismos, esta pesquisa observacional investigou a infraestrutura material e a adesão à higienização das mãos em unidade de terapia intensiva do sul do Brasil, em 2010. Os dados foram coletados por observação direta não participante e emprego de instrumento autoaplicável a 39 profissionais, analisados com auxílio de Teste do 2, estatística descritiva e análise de discurso quantitativa. Embora os profissionais superestimem a adesão, reconheçam a prática como relevante para a prevenção de infecções e refiram não haver fatores de impedimento, entre 1277 oportunidades observadas, a adesão foi de 28,6%, e significativamente menor antes do contato e dos procedimentos assépticos do que após o contato com o paciente. A infraestrutura apresentou-se deficiente em funcionalidade. Os resultados implicam risco para a segurança dos pacientes, sendo relevante o planejamento de ações corretivas e que promovam essa prática

HNF4alpha Dysfunction as a Molecular Rational for Cyclosporine Induced Hypertension

Induction of tolerance against grafted organs is achieved by the immunosuppressive agent cyclosporine, a prominent member of the calcineurin inhibitors. Unfortunately, its lifetime use is associated with hypertension and nephrotoxicity. Several mechanism for cyclosporine induced hypertension have been proposed, i.e. activation of the sympathetic nervous system, endothelin-mediated systemic vasoconstriction, impaired vasodilatation secondary to reduction in prostaglandin and nitric oxide, altered cytosolic calcium translocation, and activation of the renin-angiotensin system (RAS). In this regard the molecular basis for undue RAS activation and an increased signaling of the vasoactive oligopeptide angiotensin II (AngII) remain elusive. Notably, angiotensinogen (AGT) is the precursor of AngII and transcriptional regulation of AGT is controlled by the hepatic nuclear factor HNF4alpha. To better understand the molecular events associated with cyclosporine induced hypertension, we investigated the effect of cyclosporine on HNF4alpha expression and activity and searched for novel HNF4alpha target genes among members of the RAS cascade. Using bioinformatic algorithm and EMSA bandshift assays we identified angiotensin II receptor type 1 (AGTR1), angiotensin I converting enzyme (ACE), and angiotensin I converting enzyme 2 (ACE2) as genes targeted by HNF4alpha. Notably, cyclosporine represses HNF4alpha gene and protein expression and its DNA-binding activity at consensus sequences to AGT, AGTR1, ACE, and ACE2. Consequently, the gene expression of AGT, AGTR1, and ACE2 was significantly reduced as evidenced by quantitative real-time RT-PCR. While RAS is composed of a sophisticated interplay between multiple factors we propose a decrease of ACE2 to enforce AngII signaling via AGTR1 to ultimately result in vasoconstriction and hypertension. Taken collectively we demonstrate cyclosporine to repress HNF4alpha activity through calcineurin inhibitor mediated inhibition of nuclear factor of activation of T-cells (NFAT) which in turn represses HNF4alpha that leads to a disturbed balance of RAS

Detection of ALK fusion transcripts in FFPE lung cancer samples by NanoString technology

Background: ALK-rearranged lung cancers exhibit specific pathologic and clinical features and are responsive to anti-ALK therapies. Therefore, the detection of ALK-rearrangement is fundamental for personalized lung cancer therapy. Recently, new molecular techniques, such as NanoString nCounter, have been developed to detect ALK fusions with more accuracy and sensitivity. Methods: In the present study, we intended to validate a NanoString nCounter ALK-fusion panel in routine biopsies of FFPE lung cancer patients. A total of 43 samples were analyzed, 13 ALK-positive and 30 ALK-negative, as previously detected by FISH and/or immunohistochemistry. Results: The NanoString panel detected the presence of the EML4-ALK, KIF5B-ALK and TFG-ALK fusion variants. We observed that all the 13 ALK-positive cases exhibited genetic aberrations by the NanoString methodology. Namely, six cases (46.15%) presented EML-ALK variant 1, two (15.38%) presented EML-ALK variant 2, two (15.38%) presented EML-ALK variant 3a, and three (23.07%) exhibited no variant but presented unbalanced expression between 5'/3' exons, similar to other positive samples. Importantly, for all these analyses, the initial input of RNA was 100 ng, and some cases displayed poor RNA quality measurements. Conclusions: In this study, we reported the great utility of NanoString technology in the assessment of ALK fusions in routine lung biopsies of FFPE specimens.This study was partially funded by FINEP (MCTI/FINEP/MS/SCTIE/DECIT), Brazil. BIOPLAT (1302/13).info:eu-repo/semantics/publishedVersio