Lead reduces tension development and the myosin ATPase activity of the rat right ventricular myocardium

Lead (Pb2+) poisoning causes hypertension, but little is known regarding its acute effects on cardiac contractility. To evaluate these effects, force was measured in right ventricular strips that were contracting isometrically in 45 male Wistar rats (250-300 g) before and after the addition of increasing concentrations of lead acetate (3, 7, 10, 30, 70, 100, and 300 µM) to the bath. Changes in rate of stimulation (0.1-1.5 Hz), relative potentiation after pauses of 15, 30, and 60 s, effect of Ca2+ concentration (0.62, 1.25, and 2.5 mM), and the effect of isoproterenol (20 ng/mL) were determined before and after the addition of 100 µM Pb2+. Effects on contractile proteins were evaluated after caffeine treatment using tetanic stimulation (10 Hz) and measuring the activity of the myosin ATPase. Pb2+ produced concentration-dependent force reduction, significant at concentrations greater than 30 µM. The force developed in response to increasing rates of stimulation became smaller at 0.5 and 0.8 Hz. Relative potentiation increased after 100 µM Pb2+ treatment. Extracellular Ca2+ increment and isoproterenol administration increased force development but after 100 µM Pb2+ treatment the force was significantly reduced suggesting an effect of the metal on the sarcolemmal Ca2+ influx. Concentration of 100 µM Pb2+ also reduced the peak and plateau force of tetanic contractions and reduced the activity of the myosin ATPase. Results showed that acute Pb2+ administration, although not affecting the sarcoplasmic reticulum activity, produces a concentration-dependent negative inotropic effect and reduces myosin ATPase activity. Results suggest that acute lead administration reduced myocardial contractility by reducing sarcolemmal calcium influx and the myosin ATPase activity. These results also suggest that lead exposure is hazardous and has toxicological consequences affecting cardiac muscle.

Myosin light chain kinase is necessary for post-shock mesenteric lymph drainage enhancement of vascular reactivity and calcium sensitivity in hemorrhagic-shocked rats

Vascular hyporeactivity is an important factor in irreversible shock, and post-shock mesenteric lymph (PSML) blockade improves vascular reactivity after hemorrhagic shock. This study explored the possible involvement of myosin light chain kinase (MLCK) in PSML-mediated vascular hyporeactivity and calcium desensitization. Rats were divided into sham (n=12), shock (n=18), and shock+drainage (n=18) groups. A hemorrhagic shock model (40±2 mmHg, 3 h) was established in the shock and shock+drainage groups. PSML drainage was performed from 1 to 3 h from start of hypotension in shock+drainage rats. Levels of phospho-MLCK (p-MLCK) were determined in superior mesenteric artery (SMA) tissue, and the vascular reactivity to norepinephrine (NE) and sensitivity to Ca2+ were observed in SMA rings in an isolated organ perfusion system. p-MLCK was significantly decreased in the shock group compared with the sham group, but increased in the shock+drainage group compared with the shock group. Substance P (1 nM), an agonist of MLCK, significantly elevated the decreased contractile response of SMA rings to both NE and Ca2+ at various concentrations. Maximum contractility (Emax) in the shock group increased with NE (from 0.179±0.038 to 0.440±0.177 g/mg, P<0.05) and Ca2+ (from 0.515±0.043 to 0.646±0.096 g/mg, P<0.05). ML-7 (0.1 nM), an inhibitor of MLCK, reduced the increased vascular response to NE and Ca2+ at various concentrations in the shock+drainage group (from 0.744±0.187 to 0.570±0.143 g/mg in Emax for NE and from 0.729±0.037 to 0.645±0.056 g/mg in Emax for Ca2+, P<0.05). We conclude that MLCK is an important contributor to PSML drainage, enhancing vascular reactivity and calcium sensitivity in rats with hemorrhagic shock.

Vascular O-GlcNAcylation augments reactivity to constrictor stimuli by prolonging phosphorylated levels of the myosin light chain

O-GlcNAcylation is a modification that alters the function of numerous proteins. We hypothesized that augmented O-GlcNAcylation levels enhance myosin light chain kinase (MLCK) and reduce myosin light chain phosphatase (MLCP) activity, leading to increased vascular contractile responsiveness. The vascular responses were measured by isometric force displacement. Thoracic aorta and vascular smooth muscle cells (VSMCs) from rats were incubated with vehicle or with PugNAc, which increases O-GlcNAcylation. In addition, we determined whether proteins that play an important role in the regulation of MLCK and MLCP activity are directly affected by O-GlcNAcylation. PugNAc enhanced phenylephrine (PE) responses in rat aortas (maximal effect, 14.2±2 vs 7.9±1 mN for vehicle, n=7). Treatment with an MLCP inhibitor (calyculin A) augmented vascular responses to PE (13.4±2 mN) and abolished the differences in PE-response between the groups. The effect of PugNAc was not observed when vessels were preincubated with ML-9, an MLCK inhibitor (7.3±2 vs 7.5±2 mN for vehicle, n=5). Furthermore, our data showed that differences in the PE-induced contractile response between the groups were abolished by the activator of AMP-activated protein kinase (AICAR; 6.1±2 vs 7.4±2 mN for vehicle, n=5). PugNAc increased phosphorylation of myosin phosphatase target subunit 1 (MYPT-1) and protein kinase C-potentiated inhibitor protein of 17 kDa (CPI-17), which are involved in RhoA/Rho-kinase-mediated inhibition of myosin phosphatase activity. PugNAc incubation produced a time-dependent increase in vascular phosphorylation of myosin light chain and decreased phosphorylation levels of AMP-activated protein kinase, which decreased the affinity of MLCK for Ca2+/calmodulin. Our data suggest that proteins that play an important role in the regulation of MLCK and MLCP activity are directly affected by O-GlcNAcylation, favoring vascular contraction.

Achados otoneurológicos em indivíduos portadores de Diabetes mellitus Tipo 1

Alterações metabólicas, como acontecem no Diabetes mellitus, têm sido mencionadas no desenvolvimento e manutenção das queixas relacionadas ao aparelho vestibular e auditivo. OBJETIVO: Investigar o aparelho vestibular em uma população de indivíduos portadores de Diabetes mellitus Tipo 1. MATERIAL E MÉTODO: Foram avaliados 19 indivíduos, sendo 10 do gênero feminino (52,6%) e 9 do gênero masculino (47,3%), com idades variando de 8 a 25 anos, com diagnóstico médico de Diabetes mellitus tipo 1. Para comparação dos resultados, foi selecionado um grupo controle com outros 19 indivíduos, equiparando-se idade e sexo. O protocolo de avaliação consistiu de anamnese, inspeção otoscópica, avaliação do equilíbrio estático e dinâmico, provas cerebelares e avaliação vectoeletronistagmográfica. DESENHO CIENTÍFICO: Clínico prospectivo. RESULTADOS: Encontrou-se na amostra estudada alteração à vectoeletronistagmografia em 36,84% (n=7) dos indivíduos portadores de Diabetes mellitus Tipo 1, sendo 21,06% (n=4) Síndrome Vestibular Periférica Deficitária e 15,79% (n=3) Síndrome Vestibular Periférica Irritativa. CONCLUSÃO: Concluiu-se que indivíduos com Diabetes mellitus Tipo 1 podem ter seu aparelho vestibular comprometido, mesmo que não apresentem queixas otoneurológicas.

Estudo comparativo entre o aproveitamento escolar de alunos de escola de 1º grau e teste de inibição de emissões otoacústicas transientes

O processamento auditivo é fundamental para a cognição e pode ocasionar deficiência no aprendizado. Os portadores de sua deficiência podem ser habilitados, melhorando o desempenho escolar. É fundamental identificá-los. Dentre os que apresentam baixo custo e facilidade operacional está o exame de emissões otoacústicas. TIPO DE ESTUDO: Clínico e experimental. OBJETIVO: Estudar a relação do aproveitamento escolar com a inibição da emissão otoacústica transiente por estímulo auditivo contralateral. Material e Métodos: Foram avaliados 39 alunos, de sete a doze anos, sendo 19 (48,7%) com bom aproveitamento escolar e 20 (51,3%) com aproveitamento inadequado. Os exames emissão otoacústica com inibição contralateral foram comparados aos resultados de aproveitamento escolar. RESULTADOS: A falha da supressão da otoemissão transiente por estímulo acústico contralateral foi mais encontrada no grupo de crianças com mau aproveitamento escolar. Foi estabelecido um valor de corte de 1.6 dB SPL de redução da otoemissão que caracteriza a criança como pertencente ao grupo com mau aproveitamento com sensibilidade de 65,0%, especificidade de 72,2%, acurácia de 68,4% e valor preditivo positivo de 72,2%. CONCLUSÃO: O teste da falha da inibição contralateral da emissão otoacústica por estímulo auditivo contralateral é preditivo de transtorno do aproveitamento escolar em indivíduos de seis a doze anos de idade.

Amadurecimento e qualidade de ameixas 'Laetitia' tratadas com ácido salicílico e 1-metilciclopropeno

O objetivo deste trabalho foi avaliar o efeito da aplicação de ácido salicílico (AS), isoladamente e em combinação com o 1-metilciclopropeno (1-MCP), sobre o amadurecimento e a qualidade de ameixas 'Laetitia', armazenadas durante 40 dias, sob refrigeração (1 ± 0,1ºC / UR 92 ± 2%). Os tratamentos avaliados foram: controle; 1 mM de AS; 5 mM de AS; 10 mM de AS; e 5 mM de AS + 1-MCP (1 µL L-1). Nos frutos tratados com 10 mM de AS, verificou-se, após a saída da câmara, maior taxa de produção de etileno e incidência de manchas na epiderme, e, após três dias em condição ambiente, maior incidência de degenerescência de polpa. Em frutos tratados com 5 mM de AS, o 1-MCP proporcionou menor produção de etileno e menor incidência de degenerescência de polpa, maiores valores de atributos de textura, além de frutos menos vermelhos. O AS não apresenta efeito sobre a ocorrência de degenerescência da polpa e o amadurecimento de ameixas 'Laetitia', porém, provoca manchas na epiderme, em doses acima de 5 mM. Em frutos tratados com 5 mM de AS, a aplicação de 1-MCP atrasa o amadurecimento e mantém a qualidade da ameixa 'Laetitia'.

Antibodies against Bovine herpesvirus 1 in dairy herds in the state of Espirito Santo, Brasil

Bovine herpesvirus 1 (BoHV-1) causes major losses in worldwide livestock, affecting the respiratory and reproductive tracts of bovine. In the past decades, the number of cases in Brazil has been gradually increasing. Therefore, it is important to assess the distribution of infection in different regions of the country. In the state of Espírito Santo (ES) the BoHV 1 infection rate in dairy cattle herds is unknown. Thus, the aim of this study was to detect neutralizing antibodies against BoHV-1 in serum samples from 1,161 non-vaccinated cows from 59 dairy cattle herds in 23 municipalities of the Metropolitan, North, Northwest and South macro-regions. The identification of seropositive cows was evaluated by the virus neutralization test. The results showed that of all serum samples evaluated 775 (66.75%) had neutralizing antibodies against BoHV-1. Moreover, all herds were found positive; however, the percentage of positive cows varied among regions; 49.06%, 62.15%, 67.21% and 80.04% for the Metropolitan, South, North and Northwest macro-regions, respectively. In this study, the results clearly indicate the dissemination of the viral agent in dairy cattle in the ES state, requiring the monitoring and control of diseases related to BoHV-1 infection.

Postharvest biological control of brown rot in peaches after cold storage preceded by preharvest chemical control 1

ABSTRACT Pathogenic fungi cause skin darkening and peach quality depreciation in post harvest. Therefore, alternative techniques to chemical treatment are necessary in order to reduce risks to human health. The aim of this study was to evaluate the effect of the application of Trichoderma harzianum in association with different fungicides applied before harvest to 'Eldorado' peaches for brown rot control and other quality parameters during storage. The treatments consisted of five preharvest fungicide applications (control, captan, iprodione, iminoctadine and tebuconazole) associated with postharvest application of T. harzianum, after cold storage (with and without application), in three evaluation times (zero, two and four days at 20 °C), resulting in a 5x2x3 factorial design. The application of T. harzianum only brought benefits to the control of brown rot when combined with the fungicide captan, at zero day shelf life. After two days, there was a greater skin darkening in peaches treated with T. harzianum compared with peaches without the treatment, except for peaches treated with the fungicide iprodione and T. harzianum The application of T. harzianum during postharvest showed no benefits for the control of brown rot, however, the association with fungicides reduced the incidence of Rhizopus stolonifer during the shelf life.

Divergência genética por análise multivariada de caracteres fenotípicos de Anacardium humile (St. Hilaire)1

RESUMO A espécie Anacardium humile (St. Hilaire) é uma fruteira nativa do Cerrado, utilizada tradicionalmente, tanto no consumo in natura como na forma de sucos, doces e geleias, mas ainda pouco explorada economicamente. Em uma área de cerrado campo sujo, com transição para cerrado campo rupestre, localizada no município de Patrocínio, Minas Gerais, Brasil, 38 genótipos de uma população dessa espécie foram estudados; com o objetivo de avaliar a diversidade fenotípica, estimada por análise multivariada, bem como características morfológicas promissoras para programas de melhoramento e conservação. Assim, o estudo mostrou variabilidade genética satisfatória dentro da população estudada, destacando-se os genótipos 6, 10, 24, 36 e 38 por elevada produção de frutos e maiores pedúnculos e por se apresentarem distantes geneticamente, sendo indicados para futuros trabalhos de melhoramento genético, para serem multiplicados e utilizados em plantios comerciais e de conservação da espécie.