Conselho Nacional de Desenvolvimento Científico e Tecnológico / Two vaccinia viruses (VACV) genetically and phenotypically divergent were isolated, in a mixed infection, from a horse lesion during an outbreak of vesicular and exanthematous disease in horses in Southern Brazil and termed Pelotas 1 (P1V) and Pelotas 2 (P2V). This thesis describes studies performed to investigate the pathogenesis of P1V and P2V infection in rabbits and guinea pigs, and to analyze the sequence of genes potentially involved in their phenotype. Chapter 1 investigated the dose-dependent susceptibility of rabbits to P1V and P2V after intranasal (IN) inoculation. Groups of weaning rabbits were inoculated with three doses of each VACV isolate (102.5 TCID50, 104.5 TCID50 e 106.5 TCID50/rabbit). The inoculation resulted in severe respiratory distress and death of most inoculated rabbits regardless the viral strain. Clinical signs started three to six days post-inoculation (pi) and culminated in death or euthanasia at days 5 to 10 pi. Viremia was detected in animals of all groups. All rabbits surviving the infection beyond day 9 pi developed neutralizing antibodies. Interstitial pneumonia, necrossupurative bronchopneumonia and diarrhea were observed in animals which died or were euthanized in extremis. These results demonstrate that P1V and P2V are virulent for rabbits and show no apparent differences in phenotype in this species. Chapter 2 describes the investigation of the susceptibility of rabbits to intradermal (ID) inoculation to VACV, in single or mixed infection. All inoculated animals developed skin lesions characterized by hyperemia, papules, vesicles pustules and ulcers. Infectious virus was detected in cutaneous lesions, lungs and intestine of animals that died during acute infection. These results demonstrate that rabbits develop cutaneous disease and systemic infection after P1V and P2V ID inoculation. Apparently, co-infected animals developed lesions more severe than those submitted to single virus infection. In chapter 3, the susceptibility and the potential of transmission of P1V and P2V by guinea pigs were investigated. For that, guinea pigs were inoculated IN with both P1V and P2V (106 TCID50/ml). The guinea pigs did not showed clinical signs but developed viremia, shed virus in secretions and seroconverted to VACV. Nevertheless, the virus was not transmitted to guinea pig sentinels maintained in close contact or when exposed to food and feces contaminated with VACV. In Chapter 4, four genes involved in virus phenotype/virulence (C7L, K2L, N1L e B1R) were submitted to nucleotide sequencing and analysis. A 15 nucleotide (nt) deletion in K2L gene was identified in P2V. The same pattern of nucleotide deletion was also detected in other genogroup 1 Brazilian VACV isolates. Point mutations were identified in K2L, C7L and N1R genes from P2V isolates when compared to P1V and to a standard VACV strain. The molecular analysis of these genes would not allow the establishment of association between the sequences/genotype and phenotype. However, this analysis indicate that the 15 nt deletion in K2L gene may be used as a molecular marker for genogroup 1 Brazilian VACV isolates. In summary, the results obtained in these studies demonstrate: i. P1V and P2V produce systemic and cutaneous disease in rabbits but they do not exhibit evident differences in virulence for rabbits; ii. Guinea pigs are susceptible to mixed P1V an P2V infection but apparently do not effectively transmit the virus; iii. P1V and P2V present some sequence differences in virulence genes and that a 15 nt deletion in K2L gene may be used as a molecular marker to distinguish between VACV genogroups. / Duas amostras de vírus vaccínia (VACV) geneticamente e fenotipicamente distintas foram isoladas de um mesmo animal em um surto de doença vesicular e exantemática em equinos no Rio Grande do Sul, e denominados Pelotas 1 (P1V) e Pelotas 2 (P2V). Esta tese descreve estudos realizados para investigar a patogenia dos isolados P1V e P2V em coelhos e cobaias, e analisar a sequência de genes potencialmente envolvidos no fenótipo desses isolados. O Capítulo 1 relata a investigação da susceptibilidade dose-dependente de coelhos ao P1V e P2V. Os animais foram inoculados pela via intranasal (IN) com três doses (102,5 DICC50, 104,5 DICC50 e 106,5DICC50/coelho) de cada um dos isolados. A inoculação resultou em enfermidade respiratória grave e morte na maioria dos coelhos, independente do isolado utilizado. Os sinais clínicos iniciaram nos dias 3 e 6 pós-inoculação (pi) e culminaram com a morte ou eutanásia dos animais, 5 a 10 dias pi. Viremia foi detectada em coelhos de todos os grupos. Anticorpos neutralizantes foram detectados em todos os animais que sobreviveram além do dia 9 pi. Pneumonia intersticial com broncopneumonia necrossupurativa e conteúdo líquido intestinal foram lesões observadas em animais inoculados com o P1V ou P2V que evoluíram para a morte ou foram motivo para a eutanásia in extremis. Esses resultados demonstram que P1V e P2V são virulentos para coelhos e não apresentam diferenças evidentes de patogenia nessa espécie. No Capítulo 2 foi investigada a susceptibilidade de coelhos após inoculação de VACV pela via intradérmica (ID). Para isso, os coelhos foram inoculados com um dos isolados ou com ambos. Todos os coelhos inoculados apresentaram lesões de pele caracterizadas por hiperemia, pápulas, vesículas, pústulas e úlceras. Excreção viral foi detectada nas lesões cutâneas e também em amostras de pulmão e intestino de animais que morreram durante a fase aguda da infecção. Os resultados desta inoculação demonstraram que coelhos desenvolvem doença cutânea e sistêmica após a inoculação ID de P1V e P2V. Algumas evidências indicam que os coelhos co-infectados desenvolveram lesões mais severas do que na infecção simples. No Capítulo 3, investigou-se a susceptibilidade e o potencial de transmissibilidade dos isolados P1V e P2V por cobaias. Para isso, cobaias foram inoculadas pela via intranasal (IN) com uma mistura dos isolados P1V e P2V (106 DICC50/ml). As cobaias não apresentaram sinais clínicos, porém excretaram o vírus nas secreções nasais, desenvolveram viremia e soroconverteram para VACV. Apesar disso, o vírus não foi transmitido a sentinelas por contato direto, indireto (aerossóis) ou por água e alimentos contaminados com fezes deliberadamente infectadas com o vírus. No Capítulo 4, quatro genes (C7L, K2L, N1L e B1R) envolvidos no fenótipo do VACV foram amplificados por PCR, sequenciados e submetidos à análise molecular. Uma deleção de 15 nucleotídeos (nt) no gene K2L foi identificada no P2V. Essa mesma deleção também foi identificada em isolados brasileiros do VACV pertencentes ao genogrupo 1. Mutações pontuais foram identificadas nos genes K2L, C7L e N1L no P2V comparando-se com o P1V e cepas de referência do VACV. A análise molecular desses genes não permite associar essas deleções/mutações presentes no P2V com o fenótipo, mas sugere que a deleção de 15 nt no gene K2L possa ser utilizado como marcador molecular de isolados de VACV do genogrupo 1. Em resumo, os resultados obtidos nesses experimentos demonstram que: i. P1V e P2V produzem doença sistêmica e cutânea em coelhos, mas não diferem fenotipicamente nessas espécies; ii. cobaias são susceptíveis à infecção mista pelo P1V e P2V, mas aparentemente não transmitem o vírus com eficiência; iii. P1V e P2V apresentam algumas diferenças em genes de virulência, sendo que a deleção de 15 nt no gene K2L pode ser utilizada como marcador de genogrupos de VACV.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The present work aims to investigate the cisplatin influence on the cochlea and the deoxiribonucleic acid (DNA) of guinea pigs. An experimental study was executed with 12 guinea pigs (Cavia porcellus). The inclusion criterion for guinea pigs in the sample was the presence of Preyer's reflex (contraction of the pinna when facing sound stimulus) and distortion product otoacoustic emissions (DPOAEs). Guinea pigs were divided into two groups: Control Group (CG) - composed by six guinea pigs, to which it was administrated physiological solution of sodium chloride 0.9% during six consecutive days; Study Group (SG) - composed by six guinea pigs to which it was administrated cisplatin in six consecutive doses of 3mg/kg/day intraperitoneally. Twenty-four hours after the last cisplatin injection, guinea pigs were sacrificed, the blood sample was collected to perform the Comet Essay, and the cochleas were removed to histological analysis. When comparing the SG guinea pigs before and after the cisplatin administration, it was verified a statistically significant reduction of the amplitude of DPOAEs, mainly in the frequencies of 1000Hz to 3998Hz. After the cisplatin administration, it was certified that the amplitude of the DPOAEs frequencies of 2830Hz and 5657Hz from the SG guinea pigs suffered a statistically significant reduction compared to the CG guinea pigs. After the Cisplatin administration there were not detected any identifiable DNA changes in the Comet essay, the histological analysis showed alterations in the organ of Corti and in the spiral ganglion. Cisplatin causes alterations in the function and cochlear morphology, however, any damage to the DNA was detected. / O presente trabalho tem como objetivo verificar a influência da cisplatina sobre a cóclea e o ácido desoxirribonucleico (DNA) de cobaias. Estudo experimental executado com 12 cobaias (Cavia porcellus). O critério de inclusão de cobaias na amostra foi a presença de reflexo de Preyer (contração do pavilhão auricular frente a estímulo sonoro) e emissões otoacústicas produto de distorção (EOAPDs). As cobaias foram dividas em dois grupos: Grupo controle (GC) - composto de seis cobaias, às quais foi administrada solução fisiológica de cloreto de sódio a 0,9% por seis dias consecutivos; Grupo estudo (GE) - composto por seis cobaias, às quais foi administrada cisplatina em seis doses consecutivas de 3mg/kg/dia via intraperitoneal. Vinte e quatro horas após a última aplicação de cisplatina as cobaias foram sacrificadas, foi coletada amostra sanguínea para realização do Ensaio Cometa, e as cócleas foram removidas para análise histológica. Ao comparar-se as cobaias do GE antes e após a administração de cisplatina verificou-se redução estatisticamente significante da amplitude das EOAPDs principalmente nas frequências de 1000Hz à 3998Hz. Após a administração de cisplatina constatou-se que a amplitude das EOAPDs nas frequências de 2830Hz e 5657Hz, das cobaias do GE, sofreram redução estatisticamente significante quando comparado com as cobaias do GC. Após a administração de cisplatina não foram detectados danos genotóxicos identificáveis no Ensaio Cometa, a análise histológica mostrou alterações no órgão de Corti e gânglio espiral. A cisplatina provoca alterações na função e morfologia coclear, no entanto não foi detectado dano genotóxico.

A implantação embrionária e a placentação em cobaios são caracterizadas pela presença trofoblastos que se destacam da placenta principal, semelhantes ao trofoblasto extra-viloso de humanos. Nestes animais ultrapassam os limites, e podem ser encontrados infiltrados no profundamente no endométrio e no em ambiente externo ultrapassando aos limites da parede uterina. A cobaia desenvolve uma importante estrutura fisiológica de troca materno-embrionária, denominada de placenta vitelina invertida, definidas como membrana fetal destituída parcial ou totalmente do revestimento trofoblástico que permite a exposição do endoderma extra-embrionário em contato direto com o tecido materno. Tais características denotam um mecanismo de controle da resposta imune materna distinta dos paradigmas estabelecidos na reprodução humana e de roedores, assim como ratos e camundongos. Sendo a mais intrigante, a destituição do trofoblasto como célula da interface-materno-fetal que controla a tolerância imune-materna.No presente trabalho, procurou-se estabelecer a organização da placenta vitelina de cobaios a partir da identificação das células que compõe esta membrana extra-embrionária e identificar em que momento ocorre à remoção das células trofoblásticas, e a subseqüente forma de interação das células da placenta vitelina na interface com o tecido materno. Para tanto foram utilizados cobaias fêmeas com idade gestacional conhecida, sacrificadas para coleta de segmentos uterinos nos períodos iniciais da gestação e destinados ao processamento histotógico de embebição em parafina. Na ausência de marcadores celulares específicos conhecidos para cobaios, foram realizados testes prospectivos com reações: citoquímicas de PAS e azul de toluidina (AT; um painel de lectinas biotinadas com afinidade específica para diferentes açúcares; e imunocitoquímica para citoqueratina. As reações realizadas com PAS e AT não identificaram populações celulares com marcação seletiva. Contudo dentre as lectinas tetadas, a Erytrina cristagali lectin (ECL) apresentou reação altamente seletiva para a população de trofoblasto mural (TM) que se origina do trofectoderma, mantendo esta reatividade ao longo da gestação. Esta marcação permitiu avaliar temporal e espacialmente o destino destas células que ao longo da gestação eram mantidas como monocamada de TM revestindo externamente a placenta vitelina e, portanto, não expondo as células do endoderma parietal ou visceral ao ecido materno. Pelo acompanhamento do desenvolvimento embrionário nos cortes seriados, foi constatada no interior do blastocisto a organização de duas massas celulares internas em pólos opostos desde a fase de pré-eclosão. Uma das massas celulares constituída de embrioblastos que dará origem aos os folhetos embrionários nas fases subseqüentes, enquanto a outra formada as células tronco trofoblásticas precursora do cone ectoplacentário (CE). A cavidade da blastocele que separa estas duas massas celulares tem a sua parede revestida pelo endoderma parietal em fase tardia, após a formação da cavidade amniótica. Estes achados demonstram a pecularidade da embriogênese no cobaio, diferente daquelas descritas para humanos e outros roedores, não permitem analogias diretas, o que pode ter contribuído para o equívoco na descrição clássica da organização e constituição da placenta vitelina invertida de constituição córion-amniótica. Isto é, o trofoblasto participa da organização da placenta vitelina inicial e permanece na membrana âmnion-córion-vitelina perfazendo todo o limite do embrião ao longo da gestação. Portanto a hipótese da placenta vitelina parcial ou totalmente invertida baseada na descrição clássica em cobaios é decorrente da interpretação equivocada da embriogênese destes animais. / The guinea pig embryo implantation and placentation is characterized by trophoblast cells detaching from the main placenta in a similar way of human extra-villous trophobasts that deeply intrude inside the endometrium and sometimes also found outside the uterine wall. Furthermore, this animal also develops inverted yolk sac placenta defined as fetal membrane partially or fully devoided of trophoblast sheet that allows extra-embryonic endoderma direct exposition to the maternal environment. These characteristics denote a distinct control mechanism of maternal immune response from the established paradigm for human and rodents (rat and mouse) reproduction, being most intriguing the depriving of trophoblast as cells of maternal-fetal interface regulating the maternal immune tolerance. The present work aimed to establish the organization of guinea pig yolk sac based on identification of cell populations composing this membrane and identification if, or, when the trophoblast cells are removed from and subsequent interaction way of yolk sac cell in interface with maternal tissue. It was used pregnant guinea pig sacrificed on established gestational day to collect uterine fragments on early pregnancy stage and processed by conventional paraffin embedding. Due to absence of known specific cell markers for guinea pig, was performed the prospective evaluation using PAS and toluidine blue (TB) cytochemistry and a screening using a panel of biotinylated lectin specific for different sugars and, anti-cytokeratin. The PAS and TB staining did not identify any specific cell population, however, among the lectins used, Erytrina cristagali lectin (ECL) showed high selective labeling to the trophoblast cells originated from the trophectoderm that was kept through the gestational period. This reaction pattern was useful to evaluate chronologically and topologically the fate of this cell and confirmed the constancy of these cells layering the yolk sac placenta in contact with maternal tissue and therefore, endodermal cells were not exposed to maternal environment. Evaluation of embryo development step by step in the serial sections showed the presence of two inner cell mass in opposite sites inside the pre-hatched blastocyst. One of this, was formed with embryoblast that latter will originate the embryonic sheets and the other formed with trophoblast stem cells (ST) will originate the ectoplacental-cone. The wall of blastocele cavity separate these two inner cell mass was initially covered by a single ECL positive mural trophoblast and only later after the amniotic cavity is formed the extraembyonic endodermal cells migrate from the embryonic sheets to cover internally the blastocele cavity to organize the yolk sac placenta. These findings show the peculiarity of guinea pig embryogenesis, quite different from those described for human and rodents and therefore, does not allow direct analogy and seems to contribute in the misunderstanding of classic description of inverted yolk sac placenta and its cellular organization. It means, the trophoblast cell participates in the early organization of yolk sac placenta and remains in chorioamniotic yolk sac fetal membrane constantly limiting the embryo surface in contact with maternal environment. Therefore, the hypothesis of complete or partially inverted yolk sac placenta seems to be a miss understanding of guinea pig embryogenesis.

O treinamento físico (TF) melhora a resposta imune de indivíduos saudáveis e traz benefícios para o paciente asmático, mas seu papel na resposta alérgica é desconhecido. Objetivo: Avaliar o papel do TF de moderada intensidade na inflamação pulmonar alérgica crônica. Métodos: 54 cobaias, divididas em 4 grupos: grupo controle (C) (não sensibilizados e não treinados), grupo OVA (sensibilizados à ovalbumina (OVA) e não treinados), grupo treinamento físico (TF) (não sensibilizados e submetidos a um TF), e grupo OVA+TF (sensibilizados à OVA e submetidos a um TF). A sensibilização à OVA teve duração de 8 semanas e o programa de TF de 6 semanas iniciando 15 dias após o início da sensibilização. Cada grupo foi dividido em 2 subgrupos. No primeiro foi avaliada a inflamação pulmonar e os níveis de óxido nítrico exalado (NOex) e no segundo, a hiperresponsividade brônquica à metacolina (Mch). Resultados: A sensibilização à OVA induziu a um aumento da densidade de eosinófilos e linfócitos, expressão de IL(interleucina)-4 e IL-13 e na espessura do músculo liso na via aerea assim como espessura do epitélio comparado aos animais não-sensibilizados (p<0,05). Os animais do grupo OVA+TF apresentaram uma redução da densidade de eosinófilos, linfócitos, IL-4 e IL-13 comparado com o grupo OVA (p<0,05). Nem a sensibilização crônica a OVA ou TF influenciaram a expressão das citocinas Th1 (IL-2 e IFN-) ou a expressão das citocinas regulatórias (IL-10 e IL-1-ra) e nos níveis de NOex. Os grupos que realizaram TF tiveram aumento na espessura do epitélio quando comparados com grupos não-treinados embora não há diferença entre os grupos na avaliação da hiperresponsividade brônquica. Conclusão: Nossos resultados sugerem que o TF reduz a inflamação alérgica sem modificar a hiperresponsividade brônquica e o remodelamento das vias aéreas / Background: Aerobic training (TF) has a positive effects on health subjects and bring benefits on the immune system of asthmatic patients. However, its role on allergic immune response remains poorly understood. Objective: To evaluate the effects of TF in chronic allergic inflammation. Methods: Fiftyfour animals, divided in 4 groups: non-trained and non-sensitized (C), nonsensitized and aerobic exercise (TF), ovalbumin sensitized and non-trained (OVA), and sensitized and aerobic exercise (OVA+TF). OVA or saline sensitization was performed during 8 weeks. TF was performed in a treadmill during 6 weeks beginning in the 3rd week of sensitization. Each group were divided in two groups. In the first one, it was evaluated airway inflammation and levels of exhaleted oxide nitric (NOex), on the second, airway hyperresponsiveness to a methacholine (Mch). Results: OVA sensitization induced an increase in the eosinophils and lymphocytes counting, expression of IL-4 and IL-13 and the amount of airway smooth muscle and epithelium thickness compared to non-sensitized animals (p<0.05). Sensitized animals submitted to TF presented a reduction in the eosinophil and lymphocyte counting, expression of IL-4 and IL-13 compared with OVA group (p<0.05) but not OVA-induced changes in airway remodeling (p>0.05). Neither OVA nor TF induced any difference in the expression of Th1 (IL-2 and IFN-) and regulatory cytokine (IL-10 and IL1-ra) and the levels of NOex. Trained groups presented an increase in epithelium thickness as compared to the nontrained groups however we did not find difference between groups on hyperresponsiveness avaliation. Conclusion: Our results suggest that TF reduces allergic airway inflammation without changes in bronchial hyperresponsiveness and airway remodeling

In the heart, the existence of an electrical memory was firstly reported by Rosenbaum et al. (1982), but Rios et al. (1975) and Garcia Moreira (1977) were those that firstly described the existence of contractile memories in the amphibian myocardium. These authors developed a mathematical model for representing such phenomenon. In the present study, we aimed to characterize two kinds of contractile memories occurring in the mammalian myocardium. One of them, depresses the tissue (the depressant memory, DM) and the other one acts by stimulating it (the excitatory memory, EM). The pivotal rationale guiding this work was: when the heart is challenged by changing the environment sources like nutrients, chemicals, temperature, etc., its behavior changes in order to optimize the energy expenditure associated with its contractility. This adaptation process allows to be reached a new state of dynamic equilibrium. In order to express such behavior, the tissue creates contractile memories for adjusting the amplitude of myocardial forces. This is provided by balancing the load of DM and EM available at each myocardial beat. The expression and accumulation of these memories were studied in the guinea pig atria submitted to the experimental protocols described previously by Seed & Walker (1988), Shimizu (2000), and Conde-Garcia (not published). The expression and accumulation of myocardial memories were described by employing two static descriptors, LODMmax and LOEMmax. They stand for the maximum load of depressant memory and the maximum load of excitatory memory, respectively. Furthermore, another pair of dynamic descriptors was also used to measure the maximum rate of erasing of the depressant memory (MREDM) and the other one to measure the maximum rate of erasing of the excitatory memory (MREEM). The static descriptors represent the transference of load of both memories but the dynamic descriptors were related to the rate of erasing of such memories. Our results brought us onto the following conclusions: 1. contractile memories are a phenomenon apart from the electrical memory because rising the external potassium from 2.7 to 7.0 mM did not modify (n n = 4) LODMmax that changed from 82,09 ± 1,58 to 81,56 ± 2,01% (p > 0,05), LOEMmax from 83,36 ± 0,56 to 90,12 ± 17,92% (p > 0,05), MREDM changed from -1,36 ± 0,67 to -1,13 ± 0,42gf/s (p > 0,05), and MREED from -2,09 ± 1,65 to -1,56 ± 1,41gf/s (p > 0,05). 2. However, the expression and accumulation of DM and EM are affected by the intracellular calcium transient. The increase of extracellular calcium from 1,37 to 5,47mM (n = 3) reduced LODMmax: from 87,56 ± 2,33 to 63,83 ± 3,78% (p < 0,05); LOEMmax from 84,36 ± 0,54 to 13,91 ± 0,11% (p < 0,05); MREDM from -2,58 ± 0,71 to -1,20 ± 0,37gf/s (p < 0,05) and MREEM from -0,90 ± 0,13 to -0,34 ± 0,05 gf/s (p < 0,05). Adding 5mM cafeine to the bath solution also reduced LODMmax from 79,88 ± 3,48 to 56,68 ± 6,62% (p < 0,05); LOEMmax from 77,14 ± 1,02 to 28,54 ± 2,11% (p < 0,05); MREDM from -1,78 ± 0,50 to -0,60 ± 0,10 gf/s (p < 0,05), and MREED from -1,74 ± 0,64 to -0,33 ± 0,14 gf/s (p < 0,05); 3. In the experimental condition employed in this work, a given beat receives both depressant and excitatory information built by the last ten beats. / A capacidade de o miocárdio memorizar foi estudada inicialmente por Rosenbaum (1982), que, entre outros, relataram uma memória elétrica no coração. Todavia, Rios e cols. (1975) e Moreira (1977) foram os primeiros a descrever a existência de memória contrátil no miocárdio de anfíbio. Eles propuseram um modelo matemático para representar esse fenômeno. A nossa proposta, contudo, visou caracterizar dois tipos de memórias contráteis. Uma delas inibe o inotropismo (memória depressora, MD) e a outra, o estimula (memória excitadora, ME). A hipótese central deste trabalho propõe que, quando o coração é desafiado por um novo ambiente (nutrientes, químicos, pH etc.), ele redefine sua atividade contrátil para que possa alcançar um novo estado de equilíbrio. Para expressar tal comportamento, o miocárdio cria memórias, visando ajustar a amplitude das forças geradas. Isto se dá por meio do balanço entre a carga de MD e de ME de cada batimento. Neste trabalho, a expressão e a acumulação destas memórias foram estudadas em átrio de cobaia, utilizando-se dois descritores para cada uma delas um estático, o IKMDmax e IKMEmax, que representam o incremento máximo de carga de MD e ME, respectivamente, e outro dinâmico VmedAMD e VmedAME - que está associado à velocidade de apagamento de cada memórias. As preparações foram ensaiadas com diferentes protocolos experimentais como os descritos por Seed & Walker (1988), Shimizu, et al. (2000) e Conde-Garcia (não publicado). Os resultados mostraram que a memória contrátil difere do fenômeno relativo à memória elétrica do miocárdio, porque, elevando-se o potássio externo de 2,7 para 7,0 mM, não houve variação significativa dos descritores, pois, para n = 3, o IKMDmax passou de 82,09 ± 1,58 para 81,56 ± 2,01% (p > 0,05), o IKMEmax passou de 83,36 ± 0,56 para 90,12 ± 17,92% (p > 0,05), a VmedAMD variou de -1,36 ± 0,67 para -1,13 ± 0,42gf/s (p > 0,05) e a VmedME foi alterada de -2,09 ± 1,65 para -1,56 ± 1,41gf/s (p > 0,05). A expressão e a acumulação das memórias são fenômenos que dependem do transiente intracelular de cálcio nas células miocárdicas. A elevação do cálcio extracelular de 1,37 para 5,47mM, para n = 3, alterou o IKMDmax: de 87,56 ± 2,33 para 63,83 ± 3,78% (p < 0,05); IKMEmax: 84,36 ± 0,54 para 13,91 ± 0,11% (p < 0,05); VmedAMD: -2,58 ± 0,71 para -1,20 ± 0,37gf/s (p < 0,05) e VmedME: -0,90 ± 0,13 para -0,34 ± 0,05 gf/s (p < 0,05). A adição de 5mM de cafeína à solução controle do banho fez o IKMDmax variar de 79,88 ± 3,48 para 56,68 ± 6,62% (p < 0,05); o IKMEmax de 77,14 ± 1,02 para 28,54 ± 2,11% (p < 0,05); a VmedAMD de -1,78 ± 0,50 para -0,60 ± 0,10 gf/s (p < 0,05) e a VmedME -1,74 ± 0,64 para -0,33 ± 0,14 gf/s (p < 0,05). Nas condições experimentais deste estudo, uma dada contração recebe informações depressoras e excitadoras que foram geradas pelos últimos dez batimentos.

Our objective was to determine whether chronic vagus nerve stimulation (VNS) mitigates pressure overload (PO)-induced remodeling of the cardioneural interface. Guinea pigs (n = 48) were randomized to right or left cervical vagus (RCV or LCV) implant. After 2 wk, chronic left ventricular PO was induced by partial (15–20%) aortic constriction. Of the 31 animals surviving PO induction, 10 were randomized to RCV VNS, 9 to LCV VNS, and 12 to sham VNS. VNS was delivered at 20 Hz and 1.14 ± 0.03 mA at a 22% duty cycle. VNS commenced 10 days after PO induction and was maintained for 40 days. Time-matched controls (n = 9) were evaluated concurrently. Echocardiograms were obtained before and 50 days after PO. At termination, intracellular current-clamp recordings of intrinsic cardiac (IC) neurons were studied in vitro to determine effects of therapy on soma characteristics. Ventricular cardiomyocyte sizes were assessed with histology along with immunoblot analysis of selected proteins in myocardial tissue extracts. In sham-treated animals, PO increased cardiac output (34%, P < 0.004), as well as systolic (114%, P < 0.04) and diastolic (49%, P < 0.002) left ventricular volumes, a hemodynamic response prevented by VNS. PO-induced enhancements of IC synaptic efficacy and muscarinic sensitivity of IC neurons were mitigated by chronic VNS. Increased myocyte size, which doubled in PO (P < 0.05), was mitigated by RCV. PO hypertrophic myocardium displayed decreased glycogen synthase (GS) protein levels and accumulation of the phosphorylated (inactive) form of GS. These PO-induced changes in GS were moderated by left VNS. Chronic VNS targets IC neurons accompanying PO to obtund associated adverse cardiomyocyte remodeling.

This paper aims to determine whether chronic vagus nerve stimulation (VNS) mitigates myocardial infarction (MI)-induced remodeling of the intrinsic cardiac nervous system (ICNS), along with the cardiac tissue it regulates. Guinea pigs underwent VNS implantation on the right cervical vagus. Two weeks later, MI was produced by ligating the ventral descending coronary artery. VNS stimulation started 7 days post-MI (20 Hz, 0.9 ± 0.2 mA, 14 s on, 48 s off; VNS-MI, n = 7) and was compared with time-matched MI animals with sham VNS (MI n = 7) vs. untreated controls (n = 8). Echocardiograms were performed before and at 90 days post-MI. At termination, IC neuronal intracellular voltage recordings were obtained from whole-mount neuronal plexuses. MI increased left ventricular end systolic volume (LVESV) 30% (P = 0.027) and reduced LV ejection fraction (LVEF) 6.5% (P < 0.001) at 90 days post-MI compared with baseline. In the VNS-MI group, LVESV and LVEF did not differ from baseline. IC neurons showed depolarization of resting membrane potentials and increased input resistance in MI compared with VNS-MI and sham controls (P < 0.05). Neuronal excitability and sensitivity to norepinephrine increased in MI and VNS-MI groups compared with controls (P < 0.05). Synaptic efficacy, as determined by evoked responses to stimulating input axons, was reduced in VNS-MI compared with MI or controls (P < 0.05). VNS induced changes in myocytes, consistent with enhanced glycogenolysis, and blunted the MI-induced increase in the proapoptotic Bcl-2-associated X protein (P < 0.05). VNS mitigates MI-induced remodeling of the ICNS, correspondingly preserving ventricular function via both neural and cardiomyocyte-dependent actions.

No description available.

Mammals are born on a precocial–altricial continuum. Altricial species produce helpless neonates with closed distant organs incapable of locomotion, whereas precocial species give birth to well-developed young that possess sophisticated sensory and locomotor capabilities. Previous studies suggest that distinct patterns of cortex development differ between precocial and altricial species. This study compares patterns of neocortex neurogenesis and maturation in the precocial guinea pig and altricial dwarf rabbit, both belonging to the taxon of Glires. We show that the principal order of neurodevelopmental events is preserved in the neocortex of both species. Moreover, we show that neurogenesis starts at a later postconceptional day and takes longer in absolute gestational days in the precocial than the altricial neocortex. Intriguingly, our data indicate that the dwarf rabbit neocortex contains a higher abundance of highly proliferative basal progenitors than the guinea pig, which might underlie its higher encephalization quotient, demonstrating that the amount of neuron production is determined by complex regulation of multiple factors. Furthermore, we show that the guinea pig neocortex exhibits a higher maturation status at birth, thus providing evidence for the notions that precocial species might have acquired the morphological machinery required to attain their high functional state at birth and that brain expansion in the precocial newborn is mainly due to prenatally initiating processes of gliogenesis and neuron differentiation instead of increased neurogenesis. Together, this study reveals important insights into the timing and cellular differences that regulate mammalian brain growth and maturation and provides a better understanding of the evolution of mammalian altriciality and presocialit

Mammals are born on a precocial–altricial continuum. Altricial species produce helpless neonates with closed distant organs incapable of locomotion, whereas precocial species give birth to well-developed young that possess sophisticated sensory and locomotor capabilities. Previous studies suggest that distinct patterns of cortex development differ between precocial and altricial species. This study compares patterns of neocortex neurogenesis and maturation in the precocial guinea pig and altricial dwarf rabbit, both belonging to the taxon of Glires. We show that the principal order of neurodevelopmental events is preserved in the neocortex of both species. Moreover, we show that neurogenesis starts at a later postconceptional day and takes longer in absolute gestational days in the precocial than the altricial neocortex. Intriguingly, our data indicate that the dwarf rabbit neocortex contains a higher abundance of highly proliferative basal progenitors than the guinea pig, which might underlie its higher encephalization quotient, demonstrating that the amount of neuron production is determined by complex regulation of multiple factors. Furthermore, we show that the guinea pig neocortex exhibits a higher maturation status at birth, thus providing evidence for the notions that precocial species might have acquired the morphological machinery required to attain their high functional state at birth and that brain expansion in the precocial newborn is mainly due to prenatally initiating processes of gliogenesis and neuron differentiation instead of increased neurogenesis. Together, this study reveals important insights into the timing and cellular differences that regulate mammalian brain growth and maturation and provides a better understanding of the evolution of mammalian altriciality and presociality.