Association between levobupivacaine and pancuronium. Interference in neuromuscular transmission and blockade in rats

ABSTRACT PURPOSE: To evaluate the effects of levobupivacaine on neuromuscular transmission and neuromuscular blockade produced by pancuronium in vitro. METHODS: Thirty rats were distributed into groups (n = 5) according to the drug used alone or in combination: Group I - levobupivacaine (5 µg.mL-1); Group II - pancuronium (2 µg.mL-1); Group III - pancuronium (2 µg.mL-1) + levobupivacaine (5µg.mL-1). The following parameters were evaluated: 1) amplitude of diaphragmatic response to indirect stimulation, before and 60 minutes after the addition of levobupivacaine and pancuronium alone, and after the addition of levobupivacaine combined with pancuronium; 2) membrane potentials (MP) and miniature endplate potentials (MEPP). RESULTS: Levobupivacaine alone did not alter the amplitude of muscle response and MP. In preparations previoulsy exposed to levobupivacaine, the block with pancuronium was significantly denser (90.2 ± 15.2%), showing a significant difference (p=0.031) in comparison to the block produced by pancuronium alone (48.9% ± 9.8%). There was a decrease in the frequency and amplitude of MEPPs. CONCLUSION: Levobupivacaine potentiated the neuromuscular blockade produced by pancuronium, confirming a presynaptic action by a decrease in miniature endplate potentials.

Ventilação elétrica: indicações e aspectos técnicos do implante cirúrgico do marca-passo de estimulação diafragmática / Electric ventilation: indications for and technical aspects of diaphragm pacing stimulation surgical implantation

OBJETIVO: Pacientes com lesão medular cervical alta em geral são dependentes de ventilação mecânica, que, embora salve vidas, está associada a complicações e redução da expectativa de vida devido a infecções respiratórias. A estimulação do diafragma por marca-passo, às vezes chamada de ventilação elétrica, induz a inspiração por estimulação dos músculos inspiratórios. Nosso objetivo foi destacar as indicações e alguns aspectos da técnica cirúrgica empregada no implante laparoscópico dos eletrodos, assim como descrever cinco casos de pacientes tetraplégicos submetidos à técnica. MÉTODOS: A seleção dos pacientes envolveu estudos de condução do nervo frênico por via transcutânea para determinar se os nervos estavam preservados. A abordagem cirúrgica foi laparoscopia clássica, com quatro trocartes. A técnica foi iniciada com o mapeamento elétrico para encontrar os "pontos motores" (pontos de contração máxima do diafragma). Se o mapeamento era bem-sucedido, dois eletrodos eram implantados na face abdominal de cada lado do diafragma para estimular ramos do nervo frênico. RESULTADOS: Dos cinco pacientes, três e um, respectivamente, eram capazes de respirar somente com o uso do marca-passo por períodos superiores a 24 e 6 h, enquanto um não era capaz. CONCLUSÕES: Embora seja necessário um acompanhamento mais longo para chegar a conclusões definitivas, os resultados iniciais são promissores, pois, no momento, a maioria dos nossos pacientes pode permanecer sem ventilação mecânica por longos períodos de tempo.

OBJECTIVE: Patients with high cervical spinal cord injury are usually dependent on mechanical ventilation support, which, albeit life saving, is associated with complications and decreased life expectancy because of respiratory infections. Diaphragm pacing stimulation (DPS), sometimes referred to as electric ventilation, induces inhalation by stimulating the inspiratory muscles. Our objective was to highlight the indications for and some aspects of the surgical technique employed in the laparoscopic insertion of the DPS electrodes, as well as to describe five cases of tetraplegic patients submitted to the technique. METHODS: Patient selection involved transcutaneous phrenic nerve studies in order to determine whether the phrenic nerves were preserved. The surgical approach was traditional laparoscopy, with four ports. The initial step was electrical mapping in order to locate the "motor points" (the points at which stimulation would cause maximal contraction of the diaphragm). If the diaphragm mapping was successful, four electrodes were implanted into the abdominal surface of the diaphragm, two on each side, to stimulate the branches of the phrenic nerve. RESULTS: Of the five patients, three could breathe using DPS alone for more than 24 h, one could do so for more than 6 h, and one could not do so at all. CONCLUSIONS: Although a longer follow-up period is needed in order to reach definitive conclusions, the initial results have been promising. At this writing, most of our patients have been able to remain ventilator-free for long periods of time.

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...

Función del diafragma durante la colocación de cargas sobre el abdomen en sujetos normales / Study of diaphragmatic muscle function during abdominal weight in normal subjects

Los efectos de las cargas en el abdomen con el objeto de producir entrenamiento del diafragma, no han sido suficientemente evaluados. Estudiamos la función del diafragma durante la colocación de cargas sobre el abdomen y con cambios en el patrón respiratorio. Se estudiaron 6 voluntarios normales. Se obtuvo flujo en la boca, presión gástrica (Pga), presión esofágica (Pes), movimiento torácico (TX) y abdominal (AB), presión inspiratoria máxima (PImax) y presión transdiafragmática media (Pdi) y máxima (Pdimax). Se calculó la relación Pdi/Pdimax y el índice tensión-tiempo del diafragma (TTdi). Etapas: patrón normal (PN), patrón abdominal (PA) y carga de 1, 2, 4 y 6 kg con PN y PA. El PA fue facilitado por las cargas sobre el abdomen. Solo con 6 kg (PN y PA) la Pga a capacidad residual funcional aumentó significativamente (p 0.001). La Pdi siguió a las variaciones de la Pga y aumentó con todos los PA (p < 0.001). Con PA y carga el índice TTdi alcanzó un valor de 0.05 ± 0.02 (p < 0.001). Las cargas no aumentaron este índice más de lo que hizo el PA solo. Nuestros hallazgos sugieren que las cargas sobre el abdomen aumentan la propiocepción relacionada con los movimientos respiratorios y descenso del diafragma. Las cargas producen cambios leves en la mecánica del diafragma (en sujetos normales, 1/3 de la carga necesaria para desarrollar fatiga). En sujetos normales estos cambios parecen ser insuficientes para producir entrenamiento de los músculos respiratorios.

The effects of the abdominal weight with the intention of producing training of the diaphragm, have not been sufficiently evaluated. We studied the function of the diaphragm during the abdominal weight training and during associated changes in the respiratory pattern. Six normal volunteers were studied. Flow at the mouth at functional residual capacity (FRC) was obtained as well as gastric pressure (Pga), esophageal pressure (Pes), thoracic and abdominal movements, maximal inspiratory pressure and mean and maximal transdiaphragmatic pressure (Pdi and Pdi max). Pdi/Pdimax and the diaphragm tension-time index (TTdi) were calculated. Studied steps: normal pattern (NP), abdominal pattern (AP) and weight of 1, 2, 4 and 6 kg with NP and AP as well. We found 1) The AP was facilitated by the abdominal weight, 2) Only with 6 kg (NP and AP) the Pga at FRC increased significantly (p 0.001), 3) the Pdi followed the variations of the Pga and increased with all the AP (p < 0.001), 4) The index TTdi load reached a value of 0.05 ± 0.02 (p < 0.001). The charges did not increase this rate more than did the AP alone. Our findings suggest abdominal weight increases propioception related to the respiratory movements and descent of the diaphragm. The loads on the abdomen produce minor changes in mechanics of the diaphragm (1/3 of the load required to develop fatigue in normal subjects). Al least in normal subjects these changes appear to be insufficient to produce respiratory muscle training.

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.

The influence of lidocaine and racemic bupivacaine on neuromuscular blockade produced by rocuronium: a study in rat phrenic nerve-diaphragm preparation / Influência da lidocaína e da bupivacaína racêmica no bloqueio neuromuscular produzido pelo rocurônio: estudo em preparação nervo frênico-diafragma de rato

PURPOSE: To evaluate in vitro lidocaine and racemic bupivacaine effects in neuromuscular transmission and in neuromuscular blockade produced by rocuronium. METHODS: Rats were distributed in 5 groups (n = 5) in agreement with the studied drugs: lidocaine, racemic bupivacaine, rocuronium, separately (Groups I, II, III); rocuronium in preparations exposed to local anesthetics (Groups IV, V). The concentrations used were: 20 µg/mL, 5 µg/mL and 4 µg/mL, for lidocaine, bupivacaine and rocuronium, respectively. It was evaluated: 1) amplitude of diaphragm muscle response to indirect stimulation, before and 60 minutes after separately addition of lidocaine, racemic bupivacaine and rocuronium and the association of local anesthetics - rocuronium; 2) membrane potentials (MP) and miniature end-plate potentials (MEPP). RESULTS: Lidocaine and bupivacaine separately didn't alter the amplitude of muscle response and MP. In preparations previously exposed to lidocaine and racemic bupivacaine, the rocuronium blockade was significantly larger (90.10 ± 9.15 percent and 100 percent, respectively), in relation to the produced by rocuronium separately (73.12 ± 9.89 percent). Lidocaine caused an increase in the frequency of MEPP, being followed by blockade; racemic bupivacaine produced decrease being followed by blockade. CONCLUSIONS: Local anesthetics potentiated the blockade caused by rocuronium. The alterations of MEPP identify presynaptic action.

OBJETIVO: Avaliar in vitro os efeitos da lidocaína e bupivacaína racêmica na transmissão neuromuscular e no bloqueio neuromuscular produzido pelo rocurônio. MÉTODOS: Ratos foram distribuídos em 5 grupos (n = 5) de acordo com a droga estudada: lidocaina, bupivacaína racêmica, rocurônio, isoladamente (Grupos I, II, III); rocurõnio em preparações expostas aos anestésicos locais (Grupos IV, V). As concentrações utilizadas foram: 20 µg/mL, 5 µg/mL e 4 µg/mL, para lidocaína, bupivacaína e rocurônio, respectivamente. Avaliou-se: 1) amplitude das respostas do músculo diafragma à estimulação indireta, antes e 60 minutos após a adição da lidocaína, bupivacaína racêmica e rocurônio isoladamente e da associação anestésicos locais - rocurônio; 2) potenciais de membrana (PM) e potenciais de placa terminal em miniatura (PPTM). RESULTADOS: A lidocaína e a bupivacaína isoladamente não alteraram a amplitude das respostas musculares e os PM. Nas preparações previamente expostas a lidocaína e a bupivacaína racêmica, o bloqueio com o rocurônio foi significativamente maior (90,10 ± 9,15 por cento e 100 por cento, respectivamente), em relação ao produzido pelo rocurônio isoladamente (73,12 ± 9,89 por cento). A lidocaína causou aumento na freqüência dos PPTM, seguido de bloqueio; a bupivacaína racêmica produziu diminuição seguida de bloqueio. CONCLUSÕES: Os anestésicos locais potencializaram o bloqueio causado pelo rocurônio. As alterações do PPTM identificam ação pré-sináptica.

Central and peripheral diaphragmatic neural function in chronic obstructive pulmonary disease assessed by magnetic stimulation

In chronic obstructive pulmonary disease [COPD], the diaphragm may contribute to respiratory decompensation. Factors responsible for such impairment are complex and not completely studied. Little is known about the state of neural diaphragmatic pathways particularly in milder grades of the disease. This study aimed to evaluate the cortical and peripheral diaphragmatic neuronal function in COPD using magnetic stimulation. Twenty patients with mild/moderate COPD with mean age of 52.5 +/- 8.5 years, were included. Diaphragmatic function was assessed by transcranial magnetic stimulation of the motor cortex followed by stimulation of the cervical roots of the phrenic nerves. The following parameters were measured: diaphragmatic resting motor threshold, cortical motor evoked potential [MEP] latency and amplitude, phrenic nerve latency and amplitude and central conduction time [CMCT]. Correlations were done between different electrophysiological parameters and age, duration of illness and variables of pulmonary function testing. Compared to controls, patients demonstrated significant prolongation of cortical [n=9] and cervical [n=3] MEP latencies and CMCT [n=9], reduced phrenic nerve amplitude [n=5] and increased resting motor threshold [n=11]. Significant correlations were identified between different electrophysiological parameters and age, duration, severity of disease and degree of hypoxemia. All patients with phrenic nerve abnormalities had cortical abnormalities [n=9]. We concluded that diaphragmatic neural dysfunction occurs in early stages of COPD. Central impairment [cortico-spinal] seems to occur earlier than peripheral [neuropathy of the phrenic nerve]. This information has important clinical and therapeutic implications. It will be important to study the effect of therapies as non-invasive ventilation and specific pharmacological interventions on these changes

Morfofisiologia da inervação do diafragma de ovinos / Morphophysiology of diaphragm innervation in sheep

Thirty diaphragms of sheep of Santa Inês breed were studied regarding their origin, division and arrangement of the right and left phrenic nerves (Fde), and the participation of other nerves in the innervation of the diaphragm. By fixing and dissecting pieces, it was found that phrenic nerves (F) frequently come from the ventral branches of the 5th (C5) and 6th (C6) cervical spinal nerves (Ec), at right (46.67 percent) and at left (43.33 percent). The F often form a lumbocostal trunk, sternal branches at right (40.00 percent) and lumbar, costal and esternal branches at left (36.68 percent). The lumbar branches of F innervate frequently at left (96.67 percent) the homolateral pillar of the diaphragma, and at right (50.00 percent) they give fillets to Vena cava caudalis. The costal branches of the F innervate at left (90.00 percent) and at right (76.66 percent) the dorsal and ventral regions of the pars costalis. The sternal branches of the F innervate at right (100.00 percent) and at left (83.33 percent) the pars sternalis and the ventral region of the pars costalis at the same side. The intercostal nerves (VII to XII pairs, 63.33 percent) contribute to innervate the diaphragm of Santa Inês sheep.

Foram estudados em 30 diafragmas de ovinos da raça Santa Inês, a origem, a divisão e a distribuição dos nervos frênicos direito e esquerdo (Fde) e a participação de outros nervos na inervação do diafragma. Mediante fixação e dissecação das peças foi observado que os nervos frênicos (F) originam-se a partir dos ramos ventrais do 5º (C5) e 6º (C6) nervos espinhais cervicais (Ec) tanto à direita (46,67 por cento) como à esquerda (43,33 por cento). Os F finalizam em tronco lombocostal e ramo esternal à direita (40,00 por cento) e em ramo lombar, costal e esternal à esquerda (36,68 por cento). Os ramos lombares dos F inervam à esquerda (96,67 por cento) o pilar homolateral do diafragma e, à direita (50,00 por cento) fornecem filetes à veia caudal. Os ramos costais dos F ramificam à esquerda (90,00 por cento) e à direita (76,67 por cento) as regiões dorsal e ventral da pars costalis. Os ramos esternais dos F inervam à direita (100,00 por cento) e à esquerda (83,33 por cento) a pars sternalis e a região ventral da pars costalis do mesmo lado. Os nervos intercostais (VIII ao XII pares, 63,33 por cento) contribuem na inervação do diafragma de ovinos da raça Santa Inês.

Influence of stimulus frequency on blockade induced by pancuronium and rocuronium: study on rats phrenic nerve-diaphragm preparation

PURPOSE: To evaluate the influence of two stimulation frequencies on the installation of neuromuscular blockade produced by pancuronium and rocuronium on the rat diaphragm. METHODS: Diaphragms were submitted to an indirect frequency stimulation of 0.1 and 1Hz (Groups I and II, respectively). Subgroups were formed (n=5) according to the neuromuscular blocker employed (pancuronium-2µg/ml and rocuronium-4µg/ml). The twitch height depression was evaluated at 5, 15 and 30 minutes after adding the neuromuscular blocker. RESULTS: The decrease in twitch height was greater (p<0.01) with a frequency of 1Hz at all time periods studied both in preparations that are blocked with pancuronium and in those that are blocked with rocuronium. CONCLUSION: The frequency of stimulation interferes significantly with the installation of neuromuscular blockade produced by pancuronium and rocuronium, since the reduction in amplitude of the rat diaphragm response was greater for 1Hz frequencies, at all periods studied.

OBJETIVO: Avaliar a influência de duas freqüências de estimulação na instalação do bloqueio neuromuscular induzido por pancurônio e rocurônio em diafragma de ratos. MÉTODOS: Os diafragmas foram submetidos a uma freqüência de estimulação indireta de 0.1 e 1Hz (Grupos I e II, respectivamente). Os animais foram divididos em subgrupos (n=5) de acordo com o bloqueador neuromuscular a ser utilizado (pancurônio-2µg/mL e rocurônio-4µg/mL). A amplitude das respostas musculares foi avaliada 5, 15 e 30 minutos após a adição do bloqueador neuromuscular à preparação. RESULTS: A redução na intensidade da contração foi maior (p<0.01) com a freqüência de 1Hz em todos os tempos avaliados para as preparações contendo pancurônio e rocurônio CONCLUSION: A freqüência de estímulo interfere significativamente na instalação do bloquio neuromuscular produzido por pancurônio e rocurônio, uma vez que a redução na amplitude da resposta do diafragma foi maior para a freqüência de 1Hz em todos os períodos estudados.

Ramificação e distribuição dos nervos frênicos no músculo diafragma do gato doméstico / Ramification and distribution of the phrenic nerves in the muscle diaphragm of the domestic cat

A ramificação e a distribuição dos nervos frênicos direito e esquerdo foram estudadas em 30 músculos diafragmas de gatos domésticos sem raça definida, 7 machos e 23 fêmeas, conservados em solução de formaldeído a 10 %. Mediante fixação e dissecção, observou-se que os nervos frênicos se ramificaram para as respectivas partes carnosas do músculo diafragma, pars lumbalis, costalis e sternalis, e terminaram em maior freqüência em tronco dorsolateral e ramo ventral (63,33 %). Notou-se ainda os seguintes arranjos: ramos dorsal, lateral e ventral (25,0 %); ramo dorsal e tronco ventrolateral (6,66 %); troncos dorsolateral e ventrolateral (3,33 %); tronco dorsolateral, ramos lateral e ventral (1,66 %). Os nervos frênicos se distribuíram simetricamente em 11 exemplares (36,66 %), exibindo somente a terminação em tronco dorsolateral e ramo ventral. Os ramos dorsais inervaram a pars lumbalis (73,33 % à direita e 56,66 % à esquerda) e a pars costalis (13,33 % à direita e 10,0 % à esquerda). O ramo dorsal direito supriu a crus mediale dexter do pilar direito (100,0 %) e o ramo dorsal esquerdo inervou a crus mediale sinister do pilar direito e o pilar esquerdo (100,0 %). Os ramos laterais inervaram a pars lumbalis (23,33 % à direita e 33,33 % à esquerda), a pars costalis (96,66 % à direita e 100,0 % à esquerda) e a pars sternalis (3,33 % somente à direita). Os ramos ventrais inervaram a região ventral da pars costalis (46,66 % à direita e 43,33 % à esquerda) e a pars sternalis (96,66 % à direita e 100,0 % à esquerda). Quatro animais fêmeas (13,33 %) exibiram cruzamento de fibras provenientes do ramo ventral esquerdo para o antímero direito sendo que em um desses exemplares (3,33 %) ocorreu conexão entre o ramo ventral esquerdo e o direito.

The ramification and the distribution of the phrenic nerves right and left had been studied in 30 muscles diaphragms of unknown breed adult domestic cats, 7 males and 23 females, and were fixed in 10 % buffered formalin solution. After fixation and dissection, it was observed that the phrenic nerves ramified for the respective carnous parts of the muscle diaphragm, pars lumbalis, costalis and sternalis, and had finished in higher frequency in dorsolateral trunk and ventral branch (63.33 %). We observed the following arrangements: dorsal, ventral and lateral branches (25.0 %); dorsal branch and ventrolateral trunk (6.66 %); dorsolateral and ventrolateral trunks (3.33 %); dorsolateral trunk, lateral and ventral branches (1.66 %). The phrenic nerves had distributed symmetrically in 11 samples (36.66 %), only showing the termination in dorsolateral trunk and ventral branch. The dorsal branches supplied pars lumbalis (73.33 % to right and 56.66 % to the left) and pars costalis (13.33 % to right and 10.0 % to the left). The right dorsal branch supplied the crus mediale dexter of the right pillar (100.0 %) and the left dorsal branch supplied the crus mediale sinister of the right pillar and the left pillar (100.0 %). The lateral branches supply pars lumbalis (23.33 % to right and 33.33 % to the left), pars costalis (96.66 % to right and 100.0 % to the left) and pars sternalis (3.33 % only to the right). The ventral branches supplied the ventral region of pars costalis (46.66 % to right and 43.33 % to the left) and pars sternalis (96.66 % to right and 100.0 % to the left). Four female animals (13.33 %) had shown fibers crossing proceeding from the left ventral branch for right antimere had been that in one of these samples (3.33 %) occurred connection between the left ventral branch and the right.

A comparative study of aging of the elastic fiber system of the diaphragm and the rectus abdominis muscles in rats

In the present study the age-related changes of the striated muscle elastic fiber system were investigated in the diaphragm and rectus abdominis muscles of 1-, 4-, 8- and 18-month-old rats. The activation patterns of these muscles differ in that the diaphragm is regularly mobilized tens of times every minute during the entire life of the animal whereas the rectus abdominis, although mobilized in respiration, is much less and more irregularly activated. The elastic fibers were stained by the Verhoeff technique for mature elastic fibers. Weigert stain was used to stain mature and elaunin elastic fibers, and Weigert-oxone to stain mature, elaunin and oxytalan elastic fibers. The density of mature and elaunin elastic fibers showed a progressive increase with age, whereas the amount of oxytalan elastic fibers decreased in both diaphragm and rectus abdominis muscles and their muscular fascias. These age-related quantitative and structural changes of the elastic fiber system may reduce the viscoelastic properties of the diaphragm and rectus abdominis muscles, which may compromise the transmission of tensile muscle strength to the tendons and may affect maximum total strength

Electrophysiological and ultrastructural analysis of the neuromuscular blockade and miotoxicity induced by the Micrurus nigrocinctus snake venom

Micrurus nigrocinctus is the most abundant coral snake in Central America. The venom of this specie induced a concentration-dependent (10-20 mug/ml) depolarization in the isolated mouse phrenic nerve-diaphragm preparations incubated ate 37 degree. D-Tubocurrarine (10 mug/ml) and alpha betaungarotoxin (3-5 mug/ml) were able to partially protect against the depolarization induced by the venom (10 mug/ml), suggesting the involvement of subsynaptic cholinergic receptors. This venom (10 mug/ml) also increased the frequency and amplitude of miniature end-plate potentials (mepps) during the first 10-20 min of incubation. Subsequently, the mepps progressively decreased and disappeared after 60 min. These responses were accompanied by ultrastructural changes involving the nerve terminals, the subsynaptic junctional folds and the muscle mitochondria. The synaptic gutter was shallow and, very often, "shrunken" terminal with omega-shaped axolemmal identations and a decreased number of synaptic vecicles were present. A common finding was the presence of numerous finger-like, membrane-bounded bodies interposed between the terminal and the Schwann cells or postsynaptic sarcolemma. The preincubation of the venom with specific antivenom or the incubation of the preparations at room temperature (24-26 degree) reduced the number and intensity of the ultrastructural alterations. The last finding suggests the involvement of a enzymatic process, probably a phospholipase A2, present in the venom. There was a good correlation between the electrophysiological and ultrastructural effects induced by the venom which allow us to conclude that M. nigrocinctus venom has a presynaptic action in the initial stages of intoxication followed by sub- and postsynaptic effects, the last being the most important cause of neuromuscular blockade. A direct action of the venom on muscle fibers may also contributes to the irreversible blockade.

Morfologia e inervaçäo do diafragma de veados (Manzana americana, Manzana simplicicornis e Blastoceros bezoarticus) / Morphology and innervation of the diaphragm of deers (Manzana americana, Manzana simplicicornis and Blastoceros bezoarticus)

A morfologia e a inervaçäo do diafragma foram estudadas em três espécies de veados (Manzana americana, Manzana simplicicornis e Blastoceros bezoarticus). Os diafragmas foram fixados em soluçäo aquosa de formol a 10 por cento e o método utilizado para evidenciar a ramificaçäo e a distribuiçäo dos nervos frênicos foi o da dissecaçäo. Os resultados mostraram que os diafragmas das três espécies säo pouco espessos, possuem porçäo carnosa periférica reduzida, um centro tendíneo (folíolos) amplo e delgado e pilares robustos. Quando consideradas em conjunto as três espécies, os nervos frênicos dividem-se por trifurcaçäo (mais à direita) e por bifurcaçäo (ramo dorsal e tronco lateroventral - mais à esquerda - e ramo ventral e tronco laterodorsal - percentagens sem diferença estatisticamente significante entre os dois lados). Os ramos dorsais, laterais e ventrais destinam-se às partes lombar, costal e costal esternal respectivamente. Foram encontradas conexöes entre os filetes nervosos