Developmental neurotoxic effects of graphene oxide exposure in zebrafish larvae (Danio rerio).

Although graphene oxide (GO), a nanomaterial with hexagonal planar layer, has been widely studied due to its applications in neurobiology that include drug delivery and tissue engineering, additional studies to assess its potential toxic effects are still needed. Thus, this study evaluated the effects of GO exposure (at 5, 10, 50 or 100mg/L) during six consecutive days on mortality, hatching, spontaneous movement, heart rate, morphology, locomotion behavior, acetylcholinesterase (AChE) activity, dopamine levels and relative gene expression of developmental neurology-related genes using zebrafish larvae. In the 5mg/L dose, synapsin IIa expression up-regulation was seen concomitantly with down-regulation of dat expression, showing a potential compensatory mechanism. Moreover, the 10mg/L exposure caused an increase in heart rate, in absolute turn angle, brain cell damage and a decrease in dopamine levels. These alterations may be associated with autophagosome formation found in GO-exposed larval brain. No changes were observed on higher doses of GO exposure, probably due to nanomaterial agglomeration. Taken together, these results show that toxic effects of GO exposure are not dose-dependent, and are preeminent in lower concentrations. Additional studies are needed to deepen the specific mechanisms of GO neurotoxicity and are required to elucidate its potential biomedical use.

A Selective Association between Central and Peripheral Lithium Levels in Remitters in Bipolar Depression: A 3T-(7) Li Magnetic Resonance Spectroscopy Study.

OBJECTIVE: The objective of this study was to evaluate brain lithium levels using (7) Li magnetic resonance spectroscopy after 6 weeks of lithium therapy in bipolar depression to test the hypothesis that brain and plasma lithium are correlated. It was also tested whether responders and remitters have different pharmacokinetics, blood and brain lithium levels (ratio) compared with those presenting suboptimal antidepressant improvement. METHOD: Twenty-three patients with bipolar disorder (I and II) during depressive episodes were included and followed up for 6 weeks at the University of Sao Paulo using flexible dose of lithium (450-900 mg/day). Sixteen patients were drug-naïve. At endpoint, patients underwent a (7) Li-MRS scan and brain lithium concentrations were calculated. RESULTS: A significant association between central and peripheral lithium levels was found only in remitters (r = 0.7, P = 0.004) but not in non-remitters (r = -0.12, P = 0.76). Also, brain lithium (but not plasma) was inversely correlated with age (r = -0.46, P = 0.025). Plasma lithium did not correlate with any clinical outcome, lithium dosage or adverse effects. CONCLUSION: These findings suggest that non-remitters may not transport lithium properly to the brain, which may underlie treatment resistance to lithium in BD. Future studies with (7) Li-MRS integrated with the evaluation of blood-brain barrier transport mechanisms and longitudinal clinical outcomes in BD and aging are warranted.

Avaliação econômica da terminação de bovinos de corte em pastagem irrigada / Economic evaluation of finishing beef cattle on irrigated pasture

Foi avaliada a viabilidade econômica de três sistemas de produção por meio da mensuração dos custos de produção e da análise dos indicadores financeiros. Os três sistemas (S1, S2 e S3), situados na fronteira oeste do Rio Grande do Sul, foram avaliados no período de outubro de 2010 a outubro de 2011. A pastagem estabelecida no S1 foi de azevém (Lolium multiflorumLam)e trevo branco (Trifolium repensLam); no S2, trevo branco, trevo vermelho (Trifolium pratenseLam), cornichão (Lotus corniculatusLam), azevém e aveia preta (Avena strigosaSchreb); e no S3, azevém, cornichão, trevo branco e trevo vermelho. Os animais foram pesados na entrada e na saída das pastagens. A coleta de dados para mensuração do custo foi realizada mensalmente utilizando-se planilhas apropriadas. As produtividades foram de 369, 772 e 637kg/ha para os sistemas S1, S2 e S3, respectivamente. O S2 apresentou o maior custo total (R$ 2.101,59/ha), enquanto o menor foi obtido pelo S1 (R$ 1.626,93/ha). Na análise do custo operacional médio dos três sistemas, os itens que apresentaram maiores valores foram a implantação de pastagem, a energia elétrica, a manutenção da pastagem e a depreciação. Quanto às margens, os sistemas S1, S2 e S3 apresentaram, respectivamente, R$ 33,46, 1.162,55 e 701,01/ha para margem bruta; R$ -63,51, 955,40 e 541,05/ha para margem operacional; e R$ -583,18, 291,61 e -28,99/ha para lucro total. A rentabilidade foi de -0,47, 4,37 e 3,01% e a lucratividade foi de -5,94, 39,92 e 27,38% para o S1, S2 e S3, respectivamente. A irrigação de pastagens para terminação de bovinos é viável economicamente, no entanto a sua adoção deve sempre visar a altas produtividades zootécnicas devido ao seu elevado custo de produção.

The economic viability of three systems of production was evaluated through the measurement of costs and analysis of financial indicators. The three systems (S1, S2 and S3), located on the Western Border of Rio Grande do Sul, were evaluated from October 2010 to October 2011. The pasture established is S1 was ryegrass (Lolium multiflorum Lam) and white clover (Trifolium repens Lam), in S2, white clover, red clover (Trifolium pratense Lam), Bird's-foot trefoil (Lotus corniculatus Lam), ryegrass and oats (Avena strigosa Schreb), and in S3, ryegrass, birdsfoot trefoil, white clover and red clover. The animals were weighed before and after the pastures. Data collection was conducted monthly, using spreadsheets suitable for the measurement of production cost. Productivities were 369, 772 and 637 kg/ha for systems S1, S2 and S3, respectively. S2 had the highest total cost (R$ 2101.59/ha), while the lower was obtained by S1 (R$ 1626.93/ha). In the analysis of the average operating costs, items that showed the highest values were the establishment of pasture, electricity, maintenance of pasture and depreciation. As for margins, the systems S1, S2, and S3 were, respectively, R$ 33.46, 1162.55 and 701.01/ha for gross margin; R$ -63.51, 955.40 and 541.05 ha-1 for operating margin; and R$ -583.18, -28.99 and 291.61 ha-1 for total profit. The return was -0.47, 4.37 and 3.01% and profitability was -5.94, 39.92 and 27.38% for the S1, S2 and S3 respectively. The positive result found in the financial evaluation of the three systems studied indicates the economic viability of irrigated pasture in finishing cattle. Irrigation of pasture for finishing cattle is economically feasible, however, its application should always aim high yields husbandry due to their high cost of production.

Avaliação econômica da terminação de bovinos de corte em pastagem irrigada / Economic evaluation of finishing beef cattle on irrigated pasture

Foi avaliada a viabilidade econômica de três sistemas de produção por meio da mensuração dos custos de produção e da análise dos indicadores financeiros. Os três sistemas (S1, S2 e S3), situados na fronteira oeste do Rio Grande do Sul, foram avaliados no período de outubro de 2010 a outubro de 2011. A pastagem estabelecida no S1 foi de azevém (Lolium multiflorumLam)e trevo branco (Trifolium repensLam); no S2, trevo branco, trevo vermelho (Trifolium pratenseLam), cornichão (Lotus corniculatusLam), azevém e aveia preta (Avena strigosaSchreb); e no S3, azevém, cornichão, trevo branco e trevo vermelho. Os animais foram pesados na entrada e na saída das pastagens. A coleta de dados para mensuração do custo foi realizada mensalmente utilizando-se planilhas apropriadas. As produtividades foram de 369, 772 e 637kg/ha para os sistemas S1, S2 e S3, respectivamente. O S2 apresentou o maior custo total (R$ 2.101,59/ha), enquanto o menor foi obtido pelo S1 (R$ 1.626,93/ha). Na análise do custo operacional médio dos três sistemas, os itens que apresentaram maiores valores foram a implantação de pastagem, a energia elétrica, a manutenção da pastagem e a depreciação. Quanto às margens, os sistemas S1, S2 e S3 apresentaram, respectivamente, R$ 33,46, 1.162,55 e 701,01/ha para margem bruta; R$ -63,51, 955,40 e 541,05/ha para margem operacional; e R$ -583,18, 291,61 e -28,99/ha para lucro total. A rentabilidade foi de -0,47, 4,37 e 3,01% e a lucratividade foi de -5,94, 39,92 e 27,38% para o S1, S2 e S3, respectivamente. A irrigação de pastagens para terminação de bovinos é viável economicamente, no entanto a sua adoção deve sempre visar a altas produtividades zootécnicas devido ao seu elevado custo de produção.(AU)

The economic viability of three systems of production was evaluated through the measurement of costs and analysis of financial indicators. The three systems (S1, S2 and S3), located on the Western Border of Rio Grande do Sul, were evaluated from October 2010 to October 2011. The pasture established is S1 was ryegrass (Lolium multiflorum Lam) and white clover (Trifolium repens Lam), in S2, white clover, red clover (Trifolium pratense Lam), Bird's-foot trefoil (Lotus corniculatus Lam), ryegrass and oats (Avena strigosa Schreb), and in S3, ryegrass, birdsfoot trefoil, white clover and red clover. The animals were weighed before and after the pastures. Data collection was conducted monthly, using spreadsheets suitable for the measurement of production cost. Productivities were 369, 772 and 637 kg/ha for systems S1, S2 and S3, respectively. S2 had the highest total cost (R$ 2101.59/ha), while the lower was obtained by S1 (R$ 1626.93/ha). In the analysis of the average operating costs, items that showed the highest values were the establishment of pasture, electricity, maintenance of pasture and depreciation. As for margins, the systems S1, S2, and S3 were, respectively, R$ 33.46, 1162.55 and 701.01/ha for gross margin; R$ -63.51, 955.40 and 541.05 ha-1 for operating margin; and R$ -583.18, -28.99 and 291.61 ha-1 for total profit. The return was -0.47, 4.37 and 3.01% and profitability was -5.94, 39.92 and 27.38% for the S1, S2 and S3 respectively. The positive result found in the financial evaluation of the three systems studied indicates the economic viability of irrigated pasture in finishing cattle. Irrigation of pasture for finishing cattle is economically feasible, however, its application should always aim high yields husbandry due to their high cost of production.(AU)

Ultrastructural study of the denervated diaphragm in rats / Estudio ultraestructural del diafragma desnervado en ratas

The structural alterations that occur in the muscle fibers of denervated rat diaphragms were studied. Fifteen adult male albino rats (Rattus norvegicus) with a mean weight of 200 g and about 60 days of age were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by sectioning of the phrenic nerve and the right antimere served as control. Each antimere was divided into fragments, which were used for analysis transmission electron microscopy. During the initial phase of denervation (4 weeks), ultrastructural muscle modifications appeared in scattered fibers and in foci along these fibers. Muscle fibers with foci of less dense and loosely arranged myofibrils, disorganized Z line, displaced T tubules, and central nucleus exhibiting reentrances and fragmented aspect were observed. After 8 weeks, formation of large aggregates of small elongated mitochondria showing altered cristae, matrix inclusions and increased electron density was noted. At 12 weeks of denervation the alterations were found to be more drastic. Nuclei with internal deposits of myofibrillar or amorphous material were observed. In these fibers, vacuoles harbored enormous myeloid structures in the subsarcolemmal or intermyofibrillar region.

Fueron estudiadas las alteraciones ultra estructurales de las fibras musculares del diafragma denervado de ratas. Fueron utilizadas 15 ratas albinas (Rattus norvegicus), machos, adultas, con peso promedio de 200 g de aproximadamente 60 días de edad. Los diafragmas crónicamente denervados fueron obtenidos después de 4, 8 y 12 semanas de denervación. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en fragmentos, que fueron utilizados para el estudio en microscopia electrónica de transmisión. Durante los períodos iniciales de denervación (4 semanas), las modificaciones en la ultraestructura del músculo se disponen en fibras dispersas y en focos a lo largo de las mismas. Se observan fibras musculares con focos de miofibrillas rarefactas y laxamente dispuestas; línea Z desorganizada; túbulos T dislocados; núcleo central con aspecto fragmentado. Después de 8 semanas de denervación, se observa la formación de numeroso agregados de pequeñas mitocondrias alargadas, con alteraciones en las crestas, inclusiones en la matriz y aumento de la electrodensidad. Con 12 semanas de denervación, las alteraciones se muestran más drásticas; se observan núcleos con depósitos internos de material miofibrillar o amorfo. En estas fibras, las vacuolas presentan grandes estructuras mieloides en la región subsarcolemal o intermiofibrillar.

Morphologic alterations resulting from denervation of the diaphragm in rats / Alteraciones morfológicas resultantes de la denervación del diafragma en ratas

The morphological alterations that occur in the muscle fibers of denervated rat diaphragms were studied. Fifteen adult male albino rats (Rattus norvegicus) with a mean weight of 200 g and about 60 days of age were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by sectioning of the phrenic nerve and the right antimere served as control. Each antimere was divided into two fragments, which were used for histological (H.E.) and histoenzymological (NADH-TR and myofibrillar ATPase). After 4 weeks, denervated muscle fibers showed important light microscopic alterations: atrophy with angular profiles in cross-sections, cytoplasm containing vacuoles, enlarged interstitial space with increased connective tissue, cellular infiltration, and muscle fibers without defined contours. The most marked alterations being observed for type IIb and IIa fibers. Eight and 12 weeks after denervation, the NADH-TR reaction showed that it was impossible to characterize the muscle fibers based on their metabolic profile...

Fueron estudiadas las alteraciones morfológicas de las fibras musculares del diafragma denervado de ratas. Se utilizaron 15 ratas albinas (Rattus norvegicus) machos, adultos, con peso promedio de 200g y cerca de 60 días de edad. Se denervó el diafragma y después de 4, 8 y 12 semanas los animales fueron sacrificados. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en fragmentos, utilizados para el estudio histológico (H-E), histoenzimológico (NADH-TR y ATPasa miofibrilar). Después de 4 semanas las fibras musculares denervadas presentaron alteraciones importantes en lamicroscopía de luz: atrofia con perfiles angulados en secciones transversales; citoplasma con vacuolas; aumento del espacio intersticial con aumento de tejido conjuntivo; infiltración celular y fibras musculares sin contornos definidos, siendo las alteraciones más marcadas en las fibras tipo IIb e IIa. Después de 8 y 12 semanas de denervación la reacción para NADH-TR demuestra que es imposible caracterizar a las fibras musculares a través de su perfil metabólico...

Morphological and morphometric study of the opossum's dorsal root ganglia neurons.

The ultrastructural characteristics and the morphometric evaluation of the different types of neurons present in the dorsal root ganglia (DRG) of the South American opossum (Didelphis albiventris) were studied. Four adult male animals were used and the neurons from cervical and lumbar DRG were removed and processed for histological and transmission electron microscopy observations. The morphometric data were obtained from serial sections stained by H/E and Masson's trichrome. The number of neurons in cervical and lumbar DRG was 22 300 and 31 000, respectively. About 68% of the cervical neurons and 62.5% of the lumbar neurons presented areas up to 1300 µm(2) and were considered as the small neurons of the DRG. The ultrastructural observations revealed two morphological types of neurons: clear large neurons and dark small neurons. The nuclei of both cell types are spherical and the chromatin is disperse and rarefected. The cytoplasm of the dark small neuron is more electron dense and shows a regular distribution of small mitochondria and many rough reticulum cisterns in the periphery. A small Golgi apparatus was close to the nucleus and many disperse neurofilaments occupy most parts of the cytoplasm. Smooth reticulum cisterns are rare and lipofucsin-like inclusions are present at some points. In the clear large neurons, the organelles are homogenously scattered through the cytoplasm. The neurofilaments are close packed forming bundles and small mitochondria and rough reticulum cisterns are disperse. Lipofucsin-like inclusions are more frequent in these cells.

Morphometric and morphological analysis of neuromuscular junction alterations in the denervated rat diaphragm / Evaluación de las alteraciones morfológicas y morfométricas de las uniones neuromusculares del diafragma denervado en ratas

The morphological and structural alterations that occur in the neuromuscular junctions of the denervated rat diaphragm were studied. Fifteen adult male albino rats (Rattus norvegicus) aged about 60 days and with a mean weight of 200 g were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by section of the phrenic nerve and the right antimere was used as control. Each antimere was divided into three fragments: one was used for histochemical (nonspecific esterase) and morphometric study of neuromuscular junctions, and the other two were used for transmission and scanning electron microscopy (SEM) analysis. Histochemical analysis of the diaphragm neuromuscular junctions after denervation showed only small changes in junction morphology. However, these junctions became smaller and elongated and presented less visible contours with increasing time of denervation. Ultrastructural analysis of neuromuscular junctions after 12 weeks showed more or less organized junctional folds on the muscle fiber surface. The junctional cytoplasm exhibited important alterations such as mitochondrial degeneration and the presence of numerous filaments. SEM revealed the presence of deep primary synaptic grooves with peripheral excavations which housed the nerve terminal boutons and exhibited internally the secondary synaptic clefts present among the junctional folds of the sarcolemma. This study showed that some of the morphological changes demonstrated in other denervated striated skeletal muscles are not repeated at the same intensity or in the same temporal pattern in the rat diaphragm.

En este trabajo se estudiaron las alteraciones morfológicas y estructurales de las uniones neuromusculares en el diafragma denervado de ratas. Se utilizaron 15 ratas albinas (Rattus norvegicus), machos, adultos, con peso promedio de 200g y cerca de 60 días de edad. Los diafragmas crónicamente denervados fueron obtenidos y los animales se sacrificaron después de 4, 8 y 12 semanas de denervación. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en 3 fragmentos: uno fue utilizado para el estudio histoquímico (esterasa inespecífica) y morfométrico. Los otros dos se destinaron al estudio de microscopía electrónica de transmisión (MET) y microscopia electrónica de barrido (MEB) de las uniones neuromusculares. El estudio histoquímico de las uniones neuromusculares posterior a la denervación, muestra que la morfología de esas uniones sufre pequeñas alteraciones. Con la evolución del tiempo de denervación esas uniones muestran tamaños menores, son alargadas y con contornos menos nítidos. La ultra-estructura de las uniones neuromusculares después de 12 semanas, demostró que la superficie de la fibra muscular exhibe pliegues de unión más o menos organizados. La región del citoplasma de unión exhibe alteraciones importantes, con degeneración mitocondrial y presencia de muchos filamentos. En MEB se observa que los botones sinápticos primarios son profundos, presentan escavaciones periféricas donde estaban alojados los botones de las terminaciones nerviosas y exhiben internamente, los espacios sinápticos secundarios presentes entre los pliegues de unión del sarcolema. Este estudio mostró que algunos patrones morfológicos demostrados en otros músculos estriados esqueléticos denervados no se repiten con la misma intensidad y curso temporal en el diafragma de ratas.

Dietary diphenyl diselenide reduces the STZ-induced toxicity.

Oxidative stress is implicated in the pathogenesis of diabetes mellitus. Selenium supplementation has some benefits in experimental models of diabetes mellitus. This study evaluated whether dietary diphenyl diselenide, a simple synthetic organoselenium compound with antioxidant properties, reduces the streptozotocin (STZ)-induced toxicity. STZ-induced diabetic rats were fed with either standard and diphenyl diselenide (10 ppm) supplemented diets. In experimental trials, dietary diphenyl diselenide significantly decreased mortality rate (p<0.05) induced by STZ treatment. No correlation between this effect and glycemic levels were found. Diphenyl diselenide intake also promoted an increase in vitamin C, -SH levels (liver, kidney and blood) and in catalase (liver and kidney) activity, which were decreased in STZ-treated rats. In enzyme assays, diphenyl diselenide supplementation caused a significant improvement in platelets NTPDase and 5'-nucleotidase activities in STZ-induced diabetic rats when compared to the control and diabetic groups (p<0.05). Nevertheless, this supplementation did not modify the inhibition induced by STZ in delta-ALA-D activity. Our findings suggest that diphenyl diselenide compound showed beneficial effects against the development of diabetes by exhibiting antioxidant properties.

Human erythrocyte delta-aminolevulinate dehydratase inhibition by monosaccharides is not mediated by oxidation of enzyme sulfhydryl groups.

The heme pathway enzyme delta-aminolevulinate dehydratase is a good marker for oxidative stress and metal intoxication. This sulfhydryl enzyme is inhibited in such oxidative pathologies as lead, mercury and aluminum intoxication, exposure to selenium organic species and diabetes. Oxidative stress is a complicating factor in diabetes, inducing non-enzymatic glucose-mediated reactions that change protein structures and impair enzyme functions. We have studied the effects of high glucose, fructose and ribose concentrations on delta-ALA-D activity in vitro. These reducing sugars inhibited delta-ALA-D with efficacies in the order fructose=ribose>glucose. The possible mechanism of glucose inhibition was investigated using lysine, DTT, and t-butylamine. Oxidation of the enzyme's critical sulfhydryl groups was not involved because DTT had no effect. We concluded that high concentrations of reducing sugars or their autoxidation products inhibit delta-ALA-D by a mechanism not related to thiol oxidation. Also, we are not able to demonstrate that the formation of a Schiff base with the critical lysine residue of the enzyme is involved in the inhibition of delta-ALA-D by hexoses.

The ultrastructure of the intestinal epithelium in Oreochromis niloticus on a diet with different concentrations of zinc.

The effects of diets with variable zinc levels on the midgut epithelial cells were studied in Oreochromis niloticus L. One hundred and twenty fry of tilapia were apportioned into 4 experimental groups (I, II, III and IV groups), with 30 fish in each treatment, 5 replicate aquaria per treatment containing 6 fish each. The animals of the 4 groups were fed with isonitrogenous (30% crude protein) and isoenergetic (3000 Kcal/Kg of digestible energy) diets with increasing quantities of zinc (44.59; 149.17; 309.93; 599.67 mg Zn/kg of diet), twice a day, for 93 days. Three fish from each group were sacrificed at 36, 66 and 93 days and samples of midgut were removed for ultrastructural analysis. After 93 days of treatment, 3 animals of each experimental group were used for the analysis of zinc concentration by atomic absorption spectrophotometry. The comparative relative index (CRI) revealed that the animals in groups II, III and IV contained, respectively, 1.99%, 34.67% and 22.78% more zinc than the mean concentration in animals from group I. The ultrastructural analysis showed enterocytes with swelling of smooth surfaced endoplasmic reticulum and dilated mitochondria with variable matrix rarefaction and cristae number reduction in the fish exposed to 599.67 mg Zn/Kg of diet at 66 and 93 days of treatment.

Mesoscopy and scanning electron microscopy of the trabecular projections in the superior sagittal sinus.

The trabecular projections of the human superior sagittal sinus were classified into types and subtypes according to spatial arrangement and shape. The horizontal and vertical projections direct laminar blood flow, while the conic type, which is avalvular, protects the openings of the superior cerebral veins in the superior sagittal sinus.

Anatomical study of the diaphragm of the opossum (Didelphis albiventris).

The anatomical characteristics of the South American opossum diaphragm were described. Five male and seven female adult opossums, weighing between 700 and 1110 g, were used. Animals were killed by ether inhalation saturation. The abdominal and thoracic walls were dissected and opened, the viscerae were removed and the diaphragm anatomy was described and photographed in situ. After diaphragm removal, some dimensional data were taken and tabled. Primary branches of the phrenic nerves were dissected under a surgical microscope. The secondary branches were studied and described by transillumination after clarification in acetic acid. The opossum diaphragm is domed and has a mean area of 54.33 +/- 3.8 cm2. Well-identified costal, sternal and lumbar parts form the peripheral muscular region. The central tendinous region presents with a V-like form. Three folioles comprise the phrenic centre and present different dimensions. The caudal vena cava passes through its foramen between the ventral and right dorsal folioles. Both right and left phrenic nerves present one ventral branch and one dorsolateral trunk in 50.0% and 66.67% of the cases, respectively.

Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats.

In the present study morphological changes occurring in the neuromuscular junctions (NMJ) of the extensor digitorum longus (EDL) and soleus muscles from albino rats (Rattus norvegicus) submitted to experimental chronic alcoholism were evaluated. Seventy two male animals aged 4 months and weighing on average 400g were divided into three groups: control, alcoholic and isocaloric. Six rats from each group were anesthetized and sacrificed after 5, 10, 15 and 18 months. The NMJ did not show detectable morphological changes in either muscle after treatment when examined by light microscopy. With respect to the dimensions, statistical analysis demonstrated a tendency to a statistically significant treatment x time interaction for the length of soleus muscle NMJ. The ultrastructural study, however, revealed that the NMJ of the soleus muscle of animals submitted to 18 months of experimental alcoholism presented important morphological alterations. Characteristically, the NMJ of these muscles is located on an elevation on the surface of the muscle fiber, presenting a regular round, oval or elliptical shape and continuous and not very deep synaptic grooves. Approximately 30% of the NMJ of alcoholic rats are irregular in shape, with the sarcolemmal elevations typical of the synapse region being flattened on at least one side, with discontinuous synaptic grooves, and deep and punctiform contacts of the synaptic buds. These data suggest that, although skeletal muscle has a greater natural resistance against the direct or indirect effects of alcohol, some submicroscopic morphological alterations are detectable in the NMJ, especially in muscles with oxidative metabolism (soleus) following long periods of ingestion of alcohol.

Elevated serum S100B protein in drug-free bipolar patients during first manic episode: a pilot study.

S100B protein is a calcium-binding protein mostly derived from glial cells, which exerts trophic or toxic effects on neural cells depending on its concentration. Since serum S100B levels has been tested as a potential marker in neuropsychiatric disorders, and structural abnormalities on glial cells have been recently associated with bipolar disorder patients, we conducted this preliminary study to examine if S100B serum levels are altered during first manic episode. We quantitated S100B in serum of 40 subjects (20 unmedicated patients during manic episode and 20 healthy matched controls). The mean+/-S.D. values for S100B for bipolar subjects were 0.065+/-0.068 microg/l and 0.018+/-0.029 microg/l for healthy controls. Increased levels of S100B in bipolar mania was statistically significant (Wilcoxon signed ranks test, Z=-2.45, P=0.01). These preliminary findings suggest that mania may increase the levels of S100B in serum of bipolar disorder patients, which could be related to adaptative neural mechanisms in bipolar mania.

Morphological characteristics of neuromuscular junctions of the opossum (Didelphis albiventris) extraocular muscles: a scanning-electron-microscopic study.

Three types of neuromuscular junctions were described in the extraocular muscles of the opossum. The present study demonstrates the three-dimensional characteristics of these neuromuscular junctions after HCl connective tissue digestion. Adult opossum of both sexes were used and the neuromuscular junctions of the extraocular muscles were examined after removal of the intramuscular connective tissue and basal layer. This material was examined with a scanning electron microscope. Two types of 'en plaque' neuromuscular junction were described: the continuous type revealed elongated and branched primary synaptic grooves separated from each other by sarcolemma protuberances with different sizes, and the discontinuous or punctiform type which presents very shallow and discontinuous grooves when compared with the former. The multiple neuromuscular junctions were observed as two or three junctions associated with the same muscular fiber. The multiple junctions were present in thin fibers (around 11 microm caliber); the en plaque junctions were associated with large diameter fibers (around 21 microm). This study confirms and reveals the detailed morphological characteristics of the three neuromuscular junction types previously described by transmission electron microscope in the extraocular muscles of opossum.

Ultrastructural aspects of the retractor ocular bulbi muscle in the opossum (Didelphis albiventris).

In the present study the different types of muscle fibers of the retractor ocular bulbi muscle of the South American opossum were classified according to their ultrastructural characteristics. The tridimensional characteristics of the neuromuscular junctions present in this muscle were also demonstrated by scanning electron microscopy (SEM). Five adult opossums, three males and two females, were perfused with fixative solution through the left ventricle and their right retractor ocular bulbi muscle was prepared for the ultrastructural study of muscle fibers. The contralateral muscle was used for the study of neuromuscular junctions by SEM. Three types of fibers were detected, denoted 1r, 2r and 3r. Only simple neuromuscular junctions of the plate type were visualized by SEM.

Histochemical study of the extensor digitorum longus and soleus muscles in alcoholic rats.

The extensor digitorum longus (EDL) and soleus (SOL) muscle fibres from albino rats submitted to experimental chronic alcoholism were evaluated in accordance with their metabolic and morphometric profiles. Twenty-seven male animals aged 4 months and weighing approximately 400 g were used. The animals were divided into three groups: control, isocaloric and alcoholic and sacrifices were carried out after 5, 10 and 15 months. The muscles were dissected, removed, cross-sectioned in a cryostat and submitted to the NADH (nicotinamide adenine dinucleotide) reaction. The SO (slow-twitch-oxidative), FG (fast-twitch-glycolytic) and FOG (fast-twitch-oxidative-glycolytic) muscle fibre types exhibited a polygonal, triangular or rounded shape and did not present noteworthy modifications in either muscles during the study. The cross-sectional areas of the fibres from the studied muscles did not present significant differences during the observations. Fibre area behaved similarly in the alcoholic animals up to the 10th month, i.e. it was decreased, as also observed in the other groups. At 15 months, however, all fibres were increased, with a predominance of FG fibres in the SOL muscle. Changes in fibre population were observed mainly in the SOL muscle of alcoholic animals: SO fibres were initially increased in number but decreased after the 10th month, and the opposite was observed for the population of FG fibres. FOG fibres increased linearly in number throughout the experiment. The statistical analysis showed nevertheless that the fibre population and cross-sectional area changes were not significant. In the alcoholic animals quantitative variations of muscle fibres were more evident in the SOL muscle, suggesting that the SOL muscle is more sensitive to the toxic action of ethanol. The results concerning the increased fibre diameter in alcoholic animals would be associated with muscle oedema induced directly or indirectly by the ethanol.

Comparative structural analysis of neuromuscular junctions in mice at different ages.

The authors studied the histochemical and ultrastructural modifications that occur in the neuromuscular junctions (NMJ) of fibularis longus muscles of mice with an age range of 3 to 21 months. Twenty-four male and female animals were killed at 3, 5, 14 and 21 months of age: 7 of them at 3 months, 4 of them at 5 month, 9 at 14 months and 4 at 21 months. The fibularis longus muscles were processed and their NMJ examined with the transmission electron microscope. The most relevant changes were associated with the degeneration and retraction of terminal axons, i.e., axons poor in synaptic vesicles with degenerated mitochondria, and exhibiting multivesicular bodies and vacuoles; exposed and widened junctional folds and cytoplasmic processes of Schwann cells located in the synaptic gutter. The presence of lysosomes or lipofuchsin in the juxtajunctional sarcoplasm was also noted. These observations suggest that the phenomena of retraction and budding occur in the NMJ with advancing age, with a predominance of events associated with degeneration, leading to profound changes in NMJ shape.

[Morphologic and histochemical aspects of the extraocular muscles of the opossum (Didelphis albiventris)]. / Aspectos morfologicos e histoquimicos de los musculos extraoculares del zorrillo (Didelphis albiventris).

The extraocular muscle fibres of the South-American opossum were determined according to their metabolic profiles using NADH-diaforase, and myofibrilar ATPase after pre-incubation in both acid (pH 4.3) and alkaline (pH 10.4) media. Three muscles were selected to study the arrangement of the fibres (obliquous dorsalis, rectus dorsalis and rectus lateralis muscles). It was demonstrated that they are organized in two layers: the orbital layer composed by small diameter fibres and the global layer with three-times thicker fibres than the former. The global layer has three fibre types: white, red and intermediate; while the orbital layer presents two fibre types, which react differently to the ATPase.