Physiological response of invasive mussel Limnoperna fortunei (Dunker, 1857) (Bivalvia: Mytilidae) submitted to transport and experimental conditions / Resposta fisiológica do mexilhão invasor Limnoperna fortunei (Dunker, 1857) (Bilvalvia: Mytilidae) submetido ao transporte e condições experimentais

Abstract Successful animal rearing under laboratory conditions for commercial processes or laboratory experiments is a complex chain that includes several stressors (e.g., sampling and transport) and incurs, as a consequence, the reduction of natural animal conditions, economic losses and inconsistent and unreliable biological results. Since the invasion of the bivalve Limnoperna fortunei (Dunker, 1857) in South America, several studies have been performed to help control and manage this fouling pest in industrial plants that use raw water. Relatively little attention has been given to the laboratory rearing procedure of L. fortunei, its condition when exposed to a stressor or its acclimation into laboratory conditions. Considering this issue, the aims of this study are to (i) investigate L. fortunei physiological responses when submitted to the depuration process and subsequent air transport (without water/dry condition) at two temperatures, based on glycogen concentrations, and (ii) monitor the glycogen concentrations in different groups when maintained for 28 days under laboratory conditions. Based on the obtained results, depuration did not affect either of the groups when they were submitted to approximately eight hours of transport. The variation in glycogen concentration among the specimens that were obtained from the field under depurated and non-depurated conditions was significant only in the first week of laboratory growth for the non-depurated group and in the second week for the depurated group. In addition, the tested temperature did not affect either of the groups that were submitted to transport. The glycogen concentrations were similar to those of the specimens that were obtained from the field in third week, which suggests that the specimens acclimated to laboratory conditions during this period of time. Thus, the results indicate that the air transport and acclimation time can be successfully incorporated into experimental studies of L. fortunei. Finally, the tolerance of L. fortunei specimens to the stressor tested herein can help us understand the invasive capacity of this mussel during the establishment process.

Resumo A criação bem sucedida de animais em condições de laboratório para processos comerciais ou experimentais é uma cadeia complexa que inclui vários fatores de estresse (ex. coleta e transporte) que tem como consequência a redução das condições naturais do animal, prejuízos econômicos e resultados biológicos inconsistentes. Desde a invasão do bivalve Limnoperna fortunei (Dunker, 1857) na América do Sul, vários estudos têm sido realizados para ajudar no controle e gestão dessa praga em plantas industriais que utilizam água. Relativamente pouca atenção tem sido dada ao processo de criação de L. fortunei em laboratório, sua condição quando exposta ao estresse e sua aclimatação a condições de laboratório. Considerando estes aspectos, os objetivos deste estudo foram: (i) investigar as respostas fisiológicas de L. fortunei submetidos ao processo de depuração e subsequente transporte (sem água/condição seca) em duas temperaturas, analisando as diferentes concentrações de glicogênio e (ii) monitorar as concentrações de glicogênio nos diferentes grupos, quando mantidos por 28 dias em condições de laboratório. Com base nos resultados obtidos, a depuração não afetou nenhum grupo quando eles foram submetidos a oito horas de transporte. A variação da concentração de glicogênio entre os espécimes do campo quando depurados e não depurados, foi significativa apenas em relação à primeira semana em laboratório para o grupo não depurado e à segunda semana para o grupo depurado. Além disto, a temperatura testada não afetou os grupos submetidos ao transporte. As concentrações de glicogénio foram semelhantes as dos espécimes do campo a partir da terceira semana, o que sugere que os espécimes estão aclimatados às condições de laboratoriais neste período de tempo. Assim, os resultados indicam que o transporte ao ar e o tempo de aclimatação podem ser incorporados com sucesso aos estudos experimentais com L. fortunei. Finalmente, o conhecimento sobre a tolerância de L. fortunei ao estresse pode ajudar a entender a capacidade invasiva deste durante o processo de estabelecimento.

Glycogen-rich clear cell carcinoma of the feline mammary gland: case report / Carcinoma de células claras rico em glicogênio da glândula mamária felina: relato de caso

The aim of this report is to describe the first histopathological, immunohistochemical, and clinical characteristics of a feline glycogen-rich clear cell carcinoma (GRCCC). A Persian queen was admitted with mammary gland tumors and underwent radical unilateral mastectomy. Overall survival was considered 33 days and death was due to clinical evolution of the disease. Microscopic evaluation demonstrated epithelial cells arranged in a predominantly solid pattern, tumor cells presented an ample, granular, and foamy clear cytoplasm, and moderate cellular pleomorfism. The presence of cytoplasmatic glycogen was confirmed through diastase digestion followed by PAS staining. Histopathological and histochemical findings lead to the diagnosis of GRCCC with regional metastases.(AU)

O objetivo deste relato de caso é descrever as características clínicas, histopatológicas e imuno-histoquímicas do primeiro carcinoma mamário de células claras rico em glicogênio em felino. Uma gata persa foi atendida com tumores na glândula mamária e foi submetida à mastectomia radical unilateral. A sobrevida livre de doença foi considerada 33 dias, e o óbito foi devido à evolução da doença. A avaliação microscópica demonstrou células epiteliais arranjadas em um padrão predominantemente sólido, as células tumorais apresentaram um citoplasma claro, amplo, granular e espumoso e pleomorfismo celular moderado. A presença do glicogênio citoplasmático foi confirmada pela digestão pela diástase, seguida da coloração de PAS. Achados histopatológicos e histoquímicos levaram ao diagnóstico de carcinoma de células claras rico em glicogênio felino com metástase regional.(AU)

Morfologia e histoquímica das glândulas sexuais acessórias de Metachirus nudicaudatus, Didelphidae-Marsupialia / Morphology and histochemistry of accessory sex glands of Metachirus nudicaudatus, Didelphidae-Marsupialia

São descritas a morfologia e a distribuição de glicogênio e mucossubstâncias na próstata e nas glândulas bulbouretrais de Metachirus nudicaudatus (Geoffroy, 1803), única espécie do gênero. A próstata é envolvida pelas túnicas adventícia e muscular, e o estroma é formado pelo conjuntivo da mucosa uretral. O parênquima é constituído pelos túbulos secretores, disseminados na mucosa uretral, e que diferem histológica e histoquimicamente nos segmentos cranial, médio e caudal. Essas diferenças morfo-histoquímicas também são observadas nas partes externa, média e interna de cada túbulo. De um modo geral, os três segmentos prostáticos secretam mucossubstâncias neutras, porém o segmento caudal produz também glicogênio. Os três pares de glândulas bulbouretrais (laterais, intermédias e mediais) do Metachirus são envolvidas por uma cápsula conjuntiva e músculo estriado esquelético. O maior par é a bulbouretral lateral que é constituída por longos túbulos secretores de mucossubstâncias neutras. As bulbouretrais intermédias são formadas por túbulos ramificados, que produzem mucossubstâncias neutras, ácidas carboxiladas e ácidas sulfatadas. Os túbulo-ácinos ramificados das bulbouretrais mediais secretam mucossubstâncias neutras. O Metachirus não possui glândulas ampulares, vesículas seminais nem glândulas de coagulação.(AU)

This paper describes the morphology and distribution of glycogen and mucous substances in the prostate and the bulbourethral glands of Metachirus nudicaudatus (Geoffroy, 1803), the only species of the genus. The prostate is surrounded by the tunica adventitia, and muscle and stroma is formed by connective urethral mucosa. The glandular parenchyma consists of secretory tubules, scattered throughout the connective tissue of the urethral mucosa which differs histologically and histochemically in cranial, middle, and caudal segments of the prostate. These morpho-histochemical differences are also observed in the outer, middle and inner parts of the tubular epithelium of each prostatic segment. In general, prostatic segments secrete neutral mucous substances, and the caudal segment also produces glycogen. The three pairs of bulbourethral glands (lateral, intermediate and medial) are surrounded by a capsule of dense connective tissue and skeletal striated muscle. The glandular parenchyma is formed by tubules or branched tubuloacinar, covered by simple epithelium which is characteristic for each pair of glands. The lateral bulbourethral glands and the medial bulbourethral glands produce neutral mucous substances and the secretion of the intermediate bulbourethral glands consists of neutral mucous substances, carboxylated acids, and sulfated acids. The M. nudicaudatus does not have ampullary glands, seminal vesicles or coagulating glands.(AU)

Seasonal variation of energy reserves and reproduction in neotropical free-tailed bats Molossus molossus (Chiroptera: Molossidae) / Variação sazonal das reservas energéticas e a reprodução do morcego neotropical de cauda livre Molossus molossus (Chiroptera: Molossidae)

Seasonal variation is a key factor regulating energy metabolism and reproduction in several mammals, including bats. This study aimed to track seasonal changes in the energy reserves of the insectivorous bat Molossus molossus associated with its reproductive cycle. Adult males were collected during the four neotropical annual seasons in Viçosa - MG, Brazil. Blood and tissues were collected for metabolic analysis and testes were removed for histology and morphometry. Our results show that liver and breast muscle glycogen concentrations were significantly lower in winter. The adiposity index was significantly higher in the fall compared to winter and spring. Seminiferous tubules were greater in diameter in animals captured in fall and winter, indicating a higher investment in spermatic production during these seasons. The percentage of Leydig cells was higher in summer compared to fall and winter. We suggest that M. molossus presents a type of seasonal reproduction with two peaks of testicular activity: one in fall, with higher sperm production (spermatogenesis), and another in summer, with higher hormone production (steroidogenesis). The metabolic pattern may be associated with reproductive events, especially due to the highest fat storage observed in the fall, which coincides with the further development of the seminiferous tubules.

A variação sazonal é um fator chave na regulação do metabolismo energético e da reprodução em vários mamíferos, incluindo os morcegos. O objetivo deste estudo foi avaliar as reservas energéticas do morcego insetívoro Molossus molossus ao longo das estações anuais associadas ao seu ciclo reprodutivo. Foram coletados machos adultos durante as quatro estações anuais na cidade de Viçosa - MG, Brasil. Para as análises metabólicas foram coletados tecidos e sangue, e os testículos foram removidos para análises histológicas e morfométricas. Os resultados mostram que as concentrações de glicogênio no fígado e músculo peitoral foram significativamente menores no inverno. O índice adiposo foi significativamente maior no outono em relação ao inverno e primavera. Os túbulos seminíferos apresentaram maiores diâmetros nos animais coletados no outono e inverno, indicando um maior investimento na produção espermática durante estas estações. A porcentagem de células de Leydig foi maior no verão em comparação com outono e inverno. Sugere-se que M. molossus apresenta um tipo de reprodução sazonal com dois picos de atividade testicular: uma no outono, com maior produção de espermatozóides (espermatogênese), e outra no verão, com maior produção de hormônio (esteroidogênese). O padrão metabólico pode estar associado a eventos reprodutivos, especialmente devido ao maior armazenamento de gordura observado no outono, que coincide com o desenvolvimento dos túbulos seminíferos.

Ischemic pre and postconditioning in skeletal muscle injury produced by ischemia and reperfusion in rats

PURPOSE: To investigate the protective effects of ischemic pre and postconditioning, as well as the association of both methods, in skeletal muscle injury produced by ischemia and reperfusion in rats. METHODS: An experimental study was designed using 40 Wistar rats divided in four groups (n=10): Control - rats submitted to ischemia for 240 minutes (min) and reperfusion for 60 min; Ischemic preconditioning (Pre) - animals submitted to three cycles of clamping and releasing the aorta for five min before being submitted to the ischemia/reperfusion procedure; Ischemic postconditioning (Post) - rats submitted to three cycles of clamping and releasing the aorta for one min after the 240-minute ischemic phase; Ischemic pre and postconditioning (Pre-post) - animals submitted to the same procedures of Pre and Post groups. Skeletal muscle injury was evaluated by measuring serum levels of aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and creatine phosphokinase (CPK); and muscular levels of malondialdehyde (MDA) and glycogen. RESULTS: AST levels were significantly higher in Pre and Pre-post groups (P<.01). There were no differences in LDH and CPK levels. Muscular MDA levels were similar. Glycogen levels were significantly higher in Pre and Pre-post groups (P<.01). CONCLUSIONS: Both preconditioning and its association with postconditioning had a protective effect by avoiding glycogen depletion in skeletal muscle in rats submitted to ischemia and reperfusion. Association of pre and postconditioning did not show advantage compared to preconditioning alone. Postconditioning alone did not show protective effect.

Short-term effects of a spinosyn's family insecticide on energy metabolism and liver morphology in frugivorous bats Artibeus lituratus (Olfers, 1818) / Efeitos em curto prazo de inseticida da família das espinosinas sobre o metabolismo energético e a morfologia hepática em morcegos frugívoros

A new class of insecticide derived from fermentation of Sacharopolyspora spinosa - spinosad, has been indicated as being of low toxicity and a natural alternative to classical pesticides. In order to elucidate several aspects related to the morphophysiological changes induced by spinosad in Artibeus lituratus, the effects of a seven-day administration on plasma glucose, glycogen, protein and lipid concentrations were evaluated, and possible changes in liver cells were examined by histological analysis. Animals were fed with spinosyn-contaminated fruit through immersion in a solution. Data reporting on metabolism revealed a decrease in hind limb muscle lipid concentration in the treated group. Morphological analysis indicated a significant increase in liver cell diameter in treated animals compared to the control group. This study indicates that spinosyn, used at its recommended dose, does not affect general energy metabolism in A. lituratus but may affect some ultrastructural characteristics of liver cells.

Uma nova classe de inseticida derivado da fermentação de Sacharopolyspora spinosa - espinosade - tem sido indicada como uma alternativa natural de baixa toxicidade aos agrotóxicos clássicos. A fim de elucidar diversos aspectos relacionados às mudanças morfofisiológicas induzidas por espinosade Artibeus lituratus, os efeitos da administração durante sete dias sobre a glicose plasmática, proteína de glicogênio, e as concentrações de lipídios foram avaliados, assim como possíveis alterações nas células do fígado foram examinadas por análise histológica. Os animais foram alimentados com frutas contaminadas com espinosade por meio de imersão em uma solução. Os dados sobre o metabolismo revelaram um decréscimo na concentração de lipídios dos músculos das patas posteriores dos animais do grupo tratado. A análise morfológica indicou um aumento significativo no diâmetro das células do fígado dos animais tratados em relação ao controle. Este estudo indica que o espinosade, utilizado na dose recomendada, não afeta o metabolismo energético em geral de A. lituratus, mas pode afetar algumas características ultraestruturais das células hepáticas.

Avaliação do glicogênio hepático correlacionado com glicose sérica em ratas castradas sob tratamento com tibolona / Evaluation of hepatic glycogen correlated with serum glucose in castrated rats under tibolone treatment

INTRODUÇÃO: O glicogênio representa a forma de armazenamento de açúcares na célula animal, sendo estocada naturalmente no hepatócito. Em sua dinâmica metabólica ocorre participação de receptores, hormônios e enzimas que mantêm e equilibram os níveis séricos desse componente. OBJETIVO: O presente estudo investigou a influência da tibolona no metabolismo glicídico hepático por meio de avaliação da presença do glicogênio hepático e níveis séricos de glicose. MATERIAL E MÉTODOS: Utilizamos ao todo 14 ratas Wistar, em menopausa cirúrgica comprovada citologicamente, tratadas diariamente com tibolona (n = 9) ou com placebo (n = 5) durante 20 semanas. Efetuou-se avaliação dos pesos do animal e do fígado e dos níveis de glicose sérica. O estudo morfológico foi realizado em cortes histológicos de tecido hepático, corados com hematoxilina e eosina (HE) e com ácido periódico de Schiff (PAS), com e sem amilase salivar. Para avaliação do glicogênio no fígado, utilizou-se grade de estudo morfológico (GEM), que delineia as regiões metabólicas e circulatórias do lóbulo hepático. RESULTADOS: O peso dos animais foi menor no Grupo Tibolona, com glicose sérica em níveis mais baixos; já o peso relativo do fígado foi significativamente maior (p < 0,001). No Grupo Controle o glicogênio apresentou distribuição heterogênea em três diferentes padrões e o Grupo Tibolona mostrou glicogênio uniforme em toda a estrutura lobular. CONCLUSÃO: A tibolona, administrada em alta dose e por tempo prolongado, determina perda de peso por deficiência alimentar, que leva a alterações nas funções hepáticas, podendo influir na glicogenólise e na gliconeogênese, com modificações do glicogênio hepático e da glicose circulante.

INTRODUCTION: Glycogen serves as glucose storage in animals and it is naturally found in hepatocytes. Receptors, hormones and enzymes, which maintain and balance serum levels of this component, participate in its metabolic dynamics. OBJECTIVE: This study investigated the influence of tibolone on the hepatic glycemic metabolism by assessing the presence of liver glycogen and serum glucose. MATERIAL AND METHODS: Fourteen castrated Wistar rats, cytologically validated as surgical menopause models, were treated with tibolone (n = 9) or placebo (n = 5) for 20 weeks. Their body and liver weight and serum glucose levels were assessed. The morphologic study was performed in histological sections of liver tissue stained with hematoxylin and eosin (HE) and periodic acid-Schiff (PAS), with and without salivary amylase. For liver glycogen analysis, a morphological study grid (MSG), which outlines the metabolic and circulatory areas in the liver lobule, was applied. RESULTS: The animals' weight was lower in the Tibolone Group, with serum glucose at lower levels, whereas the relative liver weight was significantly higher (p < 0.001). The Control Group showed heterogeneous glycogen distribution in three different patterns. The Tibolone Group presented uniform glycogen throughout the lobular structure. CONCLUSION:Tibolone administration in high doses and for a long period determines weight loss by dietary deficiency, which leads to liver function changes. Thus, it may affect glycogenolysis and gluconeogenesis with changes in liver glycogen and circulating glucose.

Changes in Glycogen and Glycosaminoglycan Levels in Hepatocytes of Iduronate-2-Sulfatase Knockout Mice before and after Recombinant Iduronate-2-Sulfatase Supplementation

PURPOSE: Mucopolysaccharidosis II (MPS II) is a lysosomal storage disorder caused by a deficiency of iduronate-2 sulfatase (IdS), which is involved in the degradation of glycosaminoglycan (GAG). In this study, the frequency of fasting hypoglycemia in patients with MPS II was investigated and changes in accumulation of glycogen and GAG in the hepatocytes of IdS-knockout (KO) mice were evaluated before and after recombinant IdS enzyme replacement therapy (ERT). MATERIALS AND METHODS: Plasma glucose levels were evaluated after an 8-hour fast in 50 patients with MPS II. The IdS-KO mice were divided into three groups (group 2; saline, group 3; 0.15 mg/kg of IdS, and group 4; 0.5 mg/kg of IdS); wild-type mice were included as controls (group 1). ERT was initiated intravenously at four weeks of age, and continued every week until 20 weeks of age. RESULTS: The mean glucose level after an 8-hour fast was 94.1 +/- 23.7 mg/dL in the patients with MPS II. Two (4%) out of 50 patients had fasting hypoglycemia. For the mice, GAG in the lysosomes nearly disappeared and glycogen particles in the cytoplasm were restored to the normal range in group 4. CONCLUSION: Glucose metabolism in patients with MPS II appeared to function well despite hepatocytic GAG accumulation and hypothetical glycogen depletion. A higher dose of IdS infusion in MPS II mice led to disappearance of lysosomal GAG and restoration of glycogen to the cytoplasm of hepatocytes.

Energy metabolism and fasting in male and female insectivorous bats Molossus molossus (Chiroptera: Molossidae) / Metabolismo energético e resposta ao jejum em morcegos insetívoros Molossus molossus (Chirpotera: Molossidae)

Metabolic adaptations induced by 24 and 48 hours of fasting were investigated in male and female insectivorous bats (Molossus molossus Pallas, 1766). For this purpose, plasma glucose, non esterified fatty acids (NEFA), glycogen, protein and lipids concentrations in liver and muscles were obtained. Data presented here demonstrate that fed bats showed plasma glucose levels similar to those reported for other mammal species. In response to fasting, glycemia was decreased only in 48 hours fasted females. Plasma NEFA levels were similar in both sexes, and did not exhibit any changes during fasting. Considering the data from energy reserve variations, fed females presented an increased content of liver glycogen as well as higher breast muscle protein and limbs lipids concentrations, compared to fed males. In response to fasting, liver and muscle glycogen levels remained unchanged. Considering protein and lipid reserves, only females showed decreased values following fasting, as seen in breast, limbs and carcass lipids and breast muscle protein reserves, but still fail to keep glucose homeostasis after 48 hours without food. Taken together, our data suggest that the energy metabolism of insectivorous bats may vary according to sexual differences, a pattern that might be associated to different reproduction investments and costs between genders.

As adaptações metabólicas induzidas pelo jejum foram investigadas em morcegos insetívoros machos e fêmeas (Molossus molossus Pallas, 1766) alimentados e submetidos ao jejum por 24 e 48 horas. Para este propósito, análises plasmáticas de glucose, ácidos graxos livres, glicogênio, proteína e lipídios do fígado e músculos foram analisados. Os dados obtidos demonstraram que o nível de glicose plasmática em morcegos alimentados foi similar ao apresentado por outras espécies de mamíferos. No entanto, em resposta ao jejum, a glicemia de fêmeas diminuiu significativamente após 48 horas, enquanto os níveis circulantes de machos permaneceram constantes. Os níveis de ácidos graxos não esterificados no plasma foram similares em ambos os sexos, e não houve mudança durante o jejum. Em relação às reservas energéticas, fêmeas alimentadas apresentaram maior teor de glicogênio no fígado, de proteína armazenada no músculo peitoral e lipídios nos músculos dos membros anteriores e posteriores, em comparação aos machos alimentados. Em resposta ao jejum, somente as fêmeas mostraram diminuição de algumas reservas energéticas, como a reserva lipídica dos músculos dos membros anteriores e posteriores, da carcaça e da reserva proteica do músculo peitoral. Apesar desta mobilização, as fêmeas, diferentemente dos machos, demonstraram uma incapacidade de manter a homeostase da glicose após 48 horas sem o alimento. Nossos dados sugerem que o metabolismo energético de morcegos insetívoros varia de acordo com o sexo, sendo que o padrão metabólico pode estar associado a diferenças de custo energético no investimento reprodutivo entre machos e fêmeas.

Diagnostic efficiency of toluidine blue with Lugol's iodine in oral premalignant and malignant lesions.

Background and Objectives: In vivo stains are prompt resources, which have emerged, in the recent years, to aid as clinical diagnostic tools in detecting early premalignant and malignant lesions. The aim of the study was to determine the diagnostic efficiency of toluidine blue with Lugol's iodine in oral premalignancies and malignancies and to evaluate the reliability of in vivo staining with toluidine blue and Lugol's iodine in the lesions at risk of malignancy. Materials and Methods: The study group comprised 30 subjects with clinically suspicious premalignant lesions and 30 subjects with clinically suspicious malignant lesions. All the lesions were stained consecutively with toluidine blue and Lugol's iodine and the dye retention were recorded with photographs. Depending on the retention of the dyes, the biopsy site was determined. The biopsy specimens were sent for histological confirmation and results were statistically analyzed. Results: The overall diagnostic accuracy of Lugol's iodine when used consecutively with toluidine blue stain in distinguishing premalignant lesions and malignant lesions was 90%. As the degree of differentiation of malignant lesions progressed toward more severity, they failed to show the retention of Lugol's iodine and the result was highly significant statistically, with a P value < 0.001. Interpretation and Conclusion: Lugol's iodine when used with toluidine blue helped in delineating the inflammatory lesions and was the mean source in determining clinically the degrees of differentiation of malignant lesions as the poorly differentiated malignant lesions without glycogen content failed to show Lugol's iodine retention. Toluidine blue with Lugol's iodine can be used as a pretherapeutic assessment of the biologic aggressiveness of the disease.

Pharmacological preconditioning with l-carnitine: relevance to myocardial hemodynamic function and glycogen and lactate content

Carnitine is a vital biologic substance facilitating fatty acids transport into mitochondria for ATP production. This study was to investigate the effects of pre-ischemic pharmacological preconditioning [PC] with L-carnitine [L-Car] on myocardial infarct size and cardiac functions in ischemic and reperfused isolated rat heart and meanwhile on left ventricular glycogen and lactate content. Isolated rat hearts were subjected to 30 min coronary artery occlusion followed by 120 min reperfusion. The hearts [n=8-12] were perfused with L-Car [0.5-5 mM] only for 15 min before to 10 min after induction of ischemia. Preconditioning of the hearts with L-Car provided concentration-dependent cardioprotection as evidenced by improved postischemic ventricular functional recovery [developed pressure, left ventricular end diastolic pressure and coronary flow rate] and reduced myocardial infarct size [p<0.001]. L-Car [2.5 mM] decreased both glycogen [p<0.001] and lactate [p<0.05] content in left ventricle during ischemia compared with the control. The results of this study demonstrate that L-Car pharmacologically precondition the hearts against ischemic and reperfusion injury in part by recovery of postischemic ventricular hemodynamic functions, depletion of glycogen and therefore reduction of lactate accumulation

Efeitos do treinamento de corrida em diferentes intensidades sobre a capacidade aeróbia e produção de lactato pelo músculo de ratos Wistar / Running training effects in different intensities on the aerobic capacity and lactate production by the muscle of Wistar rats

São raros os estudos que associam indicadores de capacidade aeróbia e os substratos produzidos pelo metabolismo muscular em ratos. Dessa forma, o objetivo do presente estudo foi verificar o efeito do treinamento de corrida em duas diferentes intensidades sobre a capacidade aeróbia e a produção de lactato pelo músculo sóleo isolado de ratos. Ratos Wistar (90 dias) tiveram a transição metabólica aeróbio-anaeróbia determinada pelo teste de máxima fase estável de lactato (MFEL). Em seguida, os ratos foram treinados 40 minutos/dia, cinco dias/semana, na velocidade equivalente à MFEL (TT) ou 5 por cento superior a essa (TS), por oito semanas. Como controles foram usados ratos mantidos sedentários (S). Ao final, todos os animais foram sacrificados para análise da produção de lactato pelo músculo sóleo isolado. No inicio do experimento, a maior parte dos animais obteve a MFEL na velocidade de 25m/min à concentração de 4,38 ± 0,22mmol/L sanguínea de lactato. Ao final do experimento, a maior parte dos ratos treinados na TT apresentou MFEL na velocidade de 25m/min, à concentração sanguínea de lactato 3,10 ± 0,27mmol/L. A maioria dos treinados TS teve MFEL na velocidade de 25m/min à concentração sanguínea de lactado de 3,36 ± 0,62mmol/L. Os sedentários mostraram a MFEL na velocidade de 20m/min à concentração sanguínea de lactato de 4,83 ± 0,67mmol/L. A produção de lactato (μmol/g.h) pelo músculo sóleo isolado foi menor no grupo TS (3,83 ± 0,62) do que nos demais (S 4,31 ± 0,58 e TT 4,71 ± 0,39). A partir dos resultados obtidos no presente estudo, pode-se concluir que o treinamento aeróbio evitou a deterioração do condicionamento aeróbio imposta pelo avanço da idade e que o treinamento físico na intensidade superior à MFEL reduziu a produção muscular de lactato.

There are few studies that associate indicators of aerobic capacity and the substrates produced by the muscular metabolism in rats. The aim of the present study was to analyze the effects of physical training in different intensities on the aerobic capacity and lactate production by the isolated soleus muscle of Wistar rats (90 days) that had the aerobic/anaerobic metabolic transition determined by the Maximal Lactate Steady State Test (MLSS). Subsequently, the rats were trained 40 minutes/day, 5 days/week, in the speed equivalent to MLSS (MT) or 5 percent above it (AT), for 8 weeks. Rats maintained sedentary (S) were used as controls. At the end, all rats were sacrificed for analysis of lactate production by the isolated soleus muscle. The main results were: in the beginning of the experiment, in most of the rats the MLSS was obtained in the speed of 25m/min, to the concentration of 4.38+0.22mmol/L of blood lactate. At the end of the experiment, most of the rats trained at the MLSS intensity presented MLSS in the speed of 25m/min, to the concentration of 3.10+0.27 mmol/L of blood lactate. Most of the animals trained above-MLSS had MLSS in the speed of 25m/min, to the concentration of 3.36+0.62 mmol/L of blood lactate. Sedentary rats showed MLSS in the speed of 20m/min to the concentration of blood lactate of 4.83+0.67mmol/L. The lactate production (μmol/g.h): S 4.31+0.58, MT 4.71+0.39, AT 3.83+0.62 was lower in the ST group., It can be concluded from the results of the present study that the aerobic training prevented the deterioration of the aerobic conditioning imposed by the age advance, and that physical training above the MLSS reduced muscle lactate production.

The effect of a low dose of clenbuterol on rat soleus muscle submitted to joint immobilization

The aim of the present study was to evaluate the effect of joint immobilization on morphometric parameters and glycogen content of soleus muscle treated with clenbuterol. Male Wistar (3-4 months old) rats were divided into 4 groups (N = 6 for each group): control, clenbuterol, immobilized, and immobilized treated with clenbuterol. Immobilization was performed with acrylic resin orthoses and 10 µg/kg body weight clenbuterol was administered subcutaneously for 7 days. The following parameters were measured the next day on soleus muscle: weight, glycogen content, cross-sectional area, and connective tissue content. The clenbuterol group showed an increase in glycogen (81.6 percent, 0.38 ± 0.09 vs 0.69 ± 0.06 mg/100 g; P < 0.05) without alteration in weight, cross-sectional area or connective tissue compared with the control group. The immobilized group showed a reduction in muscle weight (34.2 percent, 123.5 ± 5.3 vs 81.3 ± 4.6 mg; P < 0.05), glycogen content (31.6 percent, 0.38 ± 0.09 vs 0.26 ± 0.05 mg/100 mg; P < 0.05) and cross-sectional area (44.1 percent, 2574.9 ± 560.2 vs 1438.1 ± 352.2 µm²; P < 0.05) and an increase in connective tissue (216.5 percent, 8.82 ± 3.55 vs 27.92 ± 5.36 percent; P < 0.05). However, the immobilized + clenbuterol group showed an increase in weight (15.9 percent; 81.3 ± 4.6 vs 94.2 ± 4.3 mg; P < 0.05), glycogen content (92.3 percent, 0.26 ± 0.05 vs 0.50 ± 0.17 mg/100 mg; P < 0.05), and cross-sectional area (19.9 percent, 1438.1 ± 352.2 vs 1724.8 ± 365.5 µm²; P < 0.05) and a reduction in connective tissue (52.2 percent, 27.92 ± 5.36 vs 13.34 ± 6.86 percent; P < 0.05). Statistical analysis was performed using Kolmogorov-Smirnov and homoscedasticity tests. For the muscle weight and muscle glycogen content, two-way ANOVA and the Tukey test were used. For the cross-sectional area and connective tissue content, Kruskal-Wallis and Tukey tests were used. This study emphasizes the importance of anabolic pharmacological...

Mecanismos das alterações do desempenho físico na programação pelo bloqueio da prolactina ao final da lactação / Mechanisms of changes in physical performance in the programming by the blockage of prolactin at the end of lactation

Anteriormente, observamos que o uso de bromocriptina (BRO), um agonista dopaminérgico inibidor de prolactina, ao final da lactação, leva ao bloqueio da produção de leite, causando uma desnutrição moderada da prole e programando uma maior massa corporal total (MCT) e de gordura visceral (MGV) e hipofunção tireóidea na idade adulta, características que são deletérias para o adequado desempenho físico. Sendo assim, avaliamos nestes animais alguns dos mecanismos relacionados à capacidade física como o conteúdo de glicogênio (muscular e hepático), as concentrações séricas de insulina, hormônios tireóideos (HTs), a atividade da enzima glicerol-fosfato desidrogenase mitocondrial (GPDm), que é regulada por HTs, no fígado, no TAM e no músculo esquelético. Indicadores de estresse oxidativo também foram avaliados utilizando o teste de espécies reativas ao ácido tiobarbitúrico (TBARs) e capacidade antioxidante total (CAT), na prole adulta aos 90 e 180 dias de idade cujas mães receberam BRO (1 mg/dia) nos 3 dias finais da lactação. O desempenho físico foi avaliado aos 90 e 180 dias de idade pela quantificação do tempo máximo de nado (TMN) em metade dos animais de cada grupo (n=10). Os ratos foram colocados em piscina com temperatura controlada (32+-2ºC) com carga adicional presa à cauda (equivalente a 5% da MCT). Aos 90 dias a MGV foi maior no grupo BRO (+56%), enquanto que os valores de glicemia (-10%) e atividade da GPDm no músculo (-53%) e n TAM (-40%) foram menores. O conteúdo de glicogênio no músculo sóleo não apresentou diferenças em resposta ao tratamento experimental ou exercício em ambas as idades, enquanto que no EDL nós observamos uma mobilização de glicogênio nos aminais exercitados de forma similar. Os animais do grupo BRO apresentaram um maior TMN (+35%), com menor produção de lactato pós-exercício (-20%). O maior conteúdo de glicogênio hepático observado no grupo BRO aos 90 dias na condição basal (+53%), e sua maior degradação com o exercício (-57%)...

Previously, we observed that the utilization of bromocriptine (BRO) a dopaminergic agonist that inhibits prolactin, at the end of lactation causes milk production blockade, provoking a moderate malnutrition in the pups and programming a higher body mass and visceral fat mass (VFM) and thyroid hypofunction in adult age, which are deleterious conditions for adequate physical performance. Therefore, we evaluated some mechanisms related to physical performance like blood lactate, glycemia and glycogen content (muscle and liver), serum insulin and thyroid hormones (THs) concentrations, mitochondrial glycerol-phosphate dehydrogenase activity (mGPD), and enzyme regulated by thyroid hormones (THs) in liver, brown adipose tissue (BAT) and skeletal muscle. Oxidative stress indicators were also evaluated using thiobarbituric acid reactive substances (TBARs) and total antioxidant capacity (TAC) in adult pups (90 and 180 days old), whose mothers received BRO (1mg/day) at the last 3 days of lactation. Physical performance was evaluated on 90 and 180 days old animals (n=10). The rats were placed in a swimming pool with controlled temperature (89+-2ºF) with additional load attached to the tail equivalent to 5% of body mass (BM). VFM was higher in 90 days old BRO group (+56%), while glycemia (-10%) and mGPD activity in muscle (-53%) and BAT (-40%) were lower. Glycogen content in soleous muscle did not present differences in response to experimental treatment or exercise in both ages, while in EDL we observed glycogen mobilization with exercise in a similar way. BRO animals presented higher MST values (+35%), with lower post-exercise lactate production (-20%). The higher hepatic glycogen content observed in BRO group at 90 days old in the basal period (+53%), and its higher degradation (-57%) together with the higher activity of mGPD in muscle in exercised animals (+172%) could contribute to the better physical performance in that age. Additionally, the higher values...

Mobilization of endogenous glycogen and trehalose of industrial yeasts

A fermentacão dos carboidratos de reserva, glicogênio e trealose é um procedimento para aumentar o nível de proteína das células de leveduras com simultâneo aumento na producão de etanol. Este trabalho estudou a cinética de degradacão do glicogênio e trealose em duas linhagens industriais de Saccharomyces cerevisiae (PE-2 e SA-1), bem como o efeito de diferentes temperaturas (38º, 40º, 42º e 44ºC) na velocidade de degradacão. A fermentacão endógena foi conduzida com suspensão de leveduras a 20 per center (m/v) em massa úmida, no vinho com 3 a 4,5 per center (v/v) de etanol. A degradacão dos carboidratos de reserva, a 40ºC, seguiu uma cinética de primeira ordem, mostrando que sua taxa é dependente da concentracão dos carboidratos na célula. A taxa especifica de degradacão (k) variou de 0,0387 a 0,0746 h-1. Em relacão a outros parâmetros analisados a 40ºC, foi observado que a viabilidade e biomassa seca e úmida foram reduzidas, enquanto a reserva de proteína celular e etanol, glicerol e nitrogênio no meio aumentaram. A degradacão do glicogênio e trealose em diferentes temperaturas (38ºC, 40ºC, 42ºC e 44ºC) mostrou que a 38ºC a taxa de degradacão foi a menor, ao passo que a partir de 42ºC ou superior, a degradacão do glicogênio não mais progrediu após poucas horas de incubacão. Portanto, do ponto de vista prático, a melhor temperatura de incubacão é em torno de 40ºC. A aplicacão da equacão de Arrheniusmostrou que as energias de ativacão de 40ºC a 42ºC foram 165,90 e 107,94 kcal.ºK-1.mol-1 para trealose e glicogênio respectivamente para a linhagem PE-2, e 190,64 e 149,87 kcal.ºK-1/mol-1, para a linhagem SA-1 respectivamente.

Changes in the hepatopancreas histology of Palaemonetes argentinus (Crustacea, Caridea) during moult

The present work describes the histological changes in the hepatopancreas of Palaemonetes argentinus during the moulting cycle. The hepatopancreas of individuals at different moult stages were dissected and studied using histological techniques. The hepatopancreas in intermoult presents four typical cell types (E, F, R and B) and follows the general plan of the rest of decapods. During early premoult (D1), there is an important secretory activity and many R-cells have several subapical vacuoles and are highly columnar. In early postmoult (A), B-cells are confined to the proximal zone of the tubules. Some of the tubules show a folded basal lamina in late premoult and postmoult. Degenerative desquamation occurs at the proximal zone of the tubules in all the stages, this zone being replenished by mitosis of E cells. Mucopolysaccharides and glycogen reserves are more abundant in premoult than in the rest of the cycle. Cellular height increased in premoult and decreased towards the intermoult, stage of more stability. R-cells are the most abundant, and F-cells do not change significantly through the cycle. The observations suggest that the hepatopancreas of P. argentinus undergoes a significant dynamic and cellular turn over rate in relation to moult.

Cardioplegia sanguínea anterógrada intermitente normotérmica e hipotérmica: estudo comparativo em coraçöes agudamente isquêmicos de coelhos / Normothermic and hypothermic intermittent anterograde blood cardioplegia: comparative study in acutely ischemic rabbit hearts

OBJETIVO: Comparar, em coraçöes agudamente isquêmicos de coelhos, estudados in vitro, o efeito bioquímico e inotrópico da infusäo anterógrada intermitente de soluçäo cardioplégica sanguínea hipercalêmica normo ou hipotérmica. MATERIAL E METODOS: Foram analisadas as concentraçöes de glicogênio, lactato e a respiraçäo mitocondrial e a Dp/dt max do ventrículo esquerdo. As cardioplegias foram infundidas a cada 20 minutos, durante 60 minutos. O estudo compreendeu as Fase I e II, cada uma envolvendo 4 grupos: Controle, sem isquemia prévia; Isquemia, submetido a estudo bioquímico após 20 minutos de isquemia normotérmica; Normotermia (cardioplegia a 37§ C) e Hipotermia (cardioplegia a 17§ C). Na Fase I, sem reperfusäo, procedeu-se apenas o estudo bioquímico. Na Fase II, com reperfusäo, fez-se o estudo funcional (dP/dt) e o metabólico ao final de reperfusço. RESULTADOS: Ao final de 20 minutos de isquemia normotérmica houve queda significativa dos níveis de glicogênio, do consumo de O2 durante o estado 3 da respiraçäo mitocondrial, da razäo do controle respiratório (RCR) e elevaçäo nos níveis de lactato, em relaçäo ao controle. A infusäo de cardioplegia hipotérmica elevou os níveis de glicogênio e restabeleceu os parâmetros da respiraçäo mitocondrial, mas näo alterou o lactato. A cardioplegia normotérmica näo alterou os níveis de glicogênio e lactato, obtidos após a isquemia aguda, mas provocou queda maior no consumo de O2 e na RCR. Na Fase II, os valores bioquímicos retornaram a valores semelhantes aos do Controle, com ambas as cardioplegias. A dP/dt max do VE näo foi significativamente diferente entre os grupos Normo e Hipotermia. CONCLUSÃO: Ambas as soluçöes cardioplégicas resultaram em alteraçöes bioquímicas similares ao final da reperfusäo e foram igualmente eficazes em preservar a funçäo do VE de coraçäo de coelhos submetidos a período prévio de 20 minutos de isquemia global, em normotermia

The importance of glucose for the freezing tolerence/intolerence of the anuran amphibians Rana catesbeiana and Bufo paracnemis

Several species of terrestrially hibernating frogs, turtles and inserts have developed mechanisms, such as increased plasma glucose, anti-freeze proteins and antioxidant enzymes that resist to freezing, for survival at subzero temperatures. In the present study, we assessed the importance of glucose to cryoresistance of two anuran amphibians: the frog Rana catesbeiana and the toad Bufo paracnemis. Both animals were exposed to -2 degrees Celsius for measurements of plasma glucose levels, liver and muscle glycogen content, haematocrit and red blood cell volume. Frogs survived cold exposure but toads did not. Blood glucose concentration increased from 40.35 + 7.25 to 131.87 + 20.72 mg/dl (P < 0.01) when the frogs were transferred from 20 to -2 degrees Celsius. Glucose accumulation in response to cold exposition in the frogs was accompanied by a decrease (P < 0.05) in liver glycogen content from 3.94 + 0.42 to 1.33 + 0.36 mg/100 mg tissue, indicating that liver carbohydrate reserves were probably the primary carbon source of glucose synthesis whereas muscle carbohydrate seems unimportant. In the toads, the cold-induced hyperglycaemia was less (P < 0.05) pronounced (from 27.25 + 1.14 to 73.72 + 13.50 mg/dl) and no significant change could be measured in liver or muscle glycogen. Cold exposition had no effect on the haematocrit of the frogs but significantly reduced (P < 0.01) the haematocrit of toads from 20.0 + 2.1 per cent to 5.8 + 1.7 per cent due to a decreased red blood cell volume (from 1532 + 63 70 728 + 87 mm3). When toads were injected with glucose, blood glucose increased to levels similar to those of frogs and haematocrit did not change, but this failed to make them cryoresistent. In conclusion, the lack of cold-induced glucose catabolism may not be the only mechanism responsible for the freeze intolerance of Bufo paracnemis, a freeze-intolerant species.

Study of the evolution of the placenta and fetal pancreas in the pathophysiology of growth retardation intrauterine due to restricted maternal diet

Intrauterine growth retard (IUGR) continues to be a significant perinatology problem at the end of this century. The nature of the etiologic agent, the time when the attack occurred during pregnancy and its duration affect the type of IUGR. Objective: To study the evolution of fetal pancreas and placenta between the 18th and 2 1st day of pregnancy in rats submitted to maternal protein-calorie restriction. Design: Randomized controlled trial on laboratory animal. Sample: Forty-one normoglycemic pregnant Wistar rats. Intervention: Rats were divided into six experimental groups according to their access to food and date of cesarean section (18th or 21 st day): control with free access to food; diet restricted to 25 per cent introduced on 1 st day of pregnancy; and diet restricted to 25 per cent after the 3rd day of pregnancy. Main measurements: Newborn weight, placenta weight, histopathological study (morphological histochemistry). Results: Maternal protein-calorie malnutrition caused intrauterine growth retard (IUGR) after the 18 th day of pregnancy. Dietary restriction did not interfere with the morphology of the fetal pancreas and theimmunohistochemical study of the placenta showed that glycogen stores were decreased between the 18 th and 21 st day in the control group and in a diet restricted to 25 per cent from the first day of pregnancy. Dietary restriction after the 3rd day of pregnancy led to low placental glycogen concentrations on the 18 th day and disappearance on the 21 st day. Conclusion: The pathophysiology of IUGR due to maternal protein-calorie restriction in rats is related to lower placental weight and low placental glycogen stores.

Biochemical alterations of the molluscan hosts tissues infected with schistosoma mansoni

As larval trematodes live within tissues of their molluscan hosts and indulge in asexual reproduction on a phenomenal scale, it is not surprising that snail tissues show biochemical changes. Levels of tissue glycogen, lactate, pyruvate and LDH[1] isoenzyme were measured in extracts of whole soft tissues of Biomphalaria alexandrina snails, specific molluscan hosts of Schistosoma mansoni. Moreover, assay of adenine nucleotides [ATP, ADP and AMP] and inorganic phosphate [Pi] was established to evaluate the adenylate energy charge [AEC] and the phosphate potential in trematode-infected snails. These parameters were measured in infected snails over four weeks, at weekly interval post exposure to S. mansoni miracidia and compared to age-matching non-exposed snails. The highest lactate/pyruvate concentration ratio with the lowest LDH[1], isoenzyme activity and glycogen level were observed two weeks post exposure. In addition, the lowest ATP content, ATP/ADP and ATP/AMP concentration ratios were also recorded at the same duration post exposure. The energy charge does not co-vary with ATP level but was slightly manipulated within the non-stressed range, while phosphate potential as a direct measure of oxygen consumption was significantly altered within 1[st], 2[nd] and 3[rd] weeks post exposure. This study clarify the most characteristic pattern of changes induced by the developing parasite. Moreover it reflects the need for an efficient glycolytic flux in the B.alexandrina snails to ensure survival and further development of S mansoni parasite