Tratamento conservativo e cirúrgico em 22 ruminantes com fraturas em membros

O objetivo do presente trabalho foi relatar os principais locais de fraturas em membros de 22 ruminantes e determinar a eficiência dos tratamentos utilizados. Foram incluídos no estudo oito caprinos, oito ovinos e seis bovinos Em casos de fraturas distais, o tratamento conservativo com imobilização foi o método de eleição. Em casos de fraturas proximais e fraturas expostas de metatarso ou metacarpo, os ruminantes foram tratados cirurgicamente. Observou-se maior frequência de fraturas envolvendo o metacarpo ou metatarso (54,5%), seguido por fraturas de tíbia (22,7%), fêmur (9%) e casos isolados de fraturas de falange medial, úmero, rádio e ulna (4,5% cada). O índice de recuperação total alcançou 95,4%. A imobilização com gesso associada à tala de Thomas, ou apenas com talas de madeira em animais jovens, foi eficiente na redução de fraturas de metacarpo, metatarso, tíbia, úmero e rádio nos ruminantes tratados. Enquanto no caso de fratura de falange medial, a imobilização com uso apenas de gesso foi suficiente para proporcionar reparação óssea adequada. A fixação esquelética externa ou interna deve ser considerada uma opção no tratamento de fraturas metacárpicas, metatársicas, tibiais e femorais em ruminantes, principalmente quando o animal for de baixo peso corporal.

Neoformação óssea e osteointegração de biomateriais micro e nanoestruturados em ovinos

A substituição de parte do tecido ósseo se faz necessária muitas vezes na rotina cirúrgica, seja em função de fraturas, neoplasias ósseas ou enfermidades ortopédicas que acarretem em perda óssea. Neste sentido, têm se buscado biomateriais capazes de promover esta substituição, evitando o uso de enxertos ou transplantes ósseos. O objetivo deste trabalho foi avaliar a capacidade osteoregenerativa de biomateriais em diferentes composições, em tíbias de ovinos. Foram utilizadas oito ovelhas mestiças texel, com 12 meses de idade e peso médio de 28,5±7,4kg. Após adequada preparação anestésica e cirúrgica, foram produzidos três defeitos ósseos na diáfise das tíbias em sua face medial, totalizando seis defeitos de 6mm cada, sendo que quatro foram preenchidos por biomateriais, e dois por fragmentos ósseos retirados do próprio animal (autocontrole). Os materiais implantados foram: hidroxiapatita (HA), tricálcio fosfato-β (TCP-β), hidroxiapatita/tricálcio fosfato-b 60/40 (HA/TCP-b 60/40) e o nanocompósito hidroxiapatita/alumina a 5% (HA/Al2O3 a 5%). Os animais foram alocados em dois grupos: Grupo 60 (n=04), em que os animais foram eutanasiados após 60 dias da colocação dos implantes e Grupo 90 (n=04), em que a eutanásia ocorreu 90 dias após a colocação dos implantes. Foram realizadas radiografias nos períodos pré-operatório, imediatamente após o procedimento e aos 30, 60 e 90 dias de pós-operatório, a fim de excluir qualquer alteração prévia ou complicação pós-operatória, capaz de comprometer o estudo. Após a eutanásia, foram coletadas as tíbias para avaliação macro e microscópica, por meio de microscopia eletrônica de varredura (MEV) e microscopia óptica. Os resultados encontrados mostraram uma boa capacidade de neoformação óssea e uma lenta absorção da HA. O TCP-β foi rapidamente absorvido e apresentou boa capacidade osteoindutiva e osteocondutiva, sendo observada neoformação óssea no interior dos grânulos deste biomaterial. O composto bifásico HA/TCP-β (60/40) apresentou o melhor resultado a longo prazo, devido ao melhor controle na solubilização e liberação dos íons cálcio e fosfatos para o meio biológico durante o processo de neoformação óssea. O biomaterial nanocompósito HA/Al2O3 a 5% não apresentou resultados promissores neste estudo, e sugerem-se novas pesquisas a fim de investigar melhor o potencial e aplicabilidade deste novo biomaterial. Conclui-se que a hidroxiapatita, o tricálcio fosfato-β e a associação HA/TCP-β (60/40) apresentam excelente capacidade de reparação óssea, podendo ser utilizados como substitutos ósseos; a associação HA/TCP-β (60/40) é o melhor dentre os biomateriais estudados, pois apresenta velocidade de absorção intermediária em relação à HA e ao TCP-β isolados, fornecendo ainda uma sustentação adequada ao tecido neoformado; o biomaterial HA/Al2O3 5% se mostrou incompatível, provocando reação de rejeição por parte do hospedeiro e com insignificante formação de tecido ósseo, sugerindo novas pesquisas acerca deste material.

Avaliação ultrassonográfica do aparelho reprodutor em serpentes vivíparas da família Boidae

A reprodução faz parte do ciclo de vida dos animais permitindo a perpetuação e a conservação das espécies. Em serpentes, existe uma escassez de informações técnicas a respeito do ciclo reprodutivo. Este estudo teve o objetivo de avaliar o aparelho reprodutivo por meio da ultrassonografia em serpentes vivíparas cativas da família Boidae, permitindo diagnosticar as diferentes fases reprodutivas. Foram avaliadas ultrassonograficamente onze serpentes adultas de quatro espécies da família Boidae: Eunectes murinus, Boa constrictor constrictor, Corallus hortulanus e Epicrates cenchria pertencentes ao acervo do Museu Biológico do Instituto Butantan, São Paulo Brasil. Para a avaliação ultrassonográfica, as serpentes foram contidas fisicamente com gancho herpetológico e depois manualmente por aproximadamente 15 minutos. A avaliação foi feita aplicando-se gel acústico sobre a pele e posicionando o transdutor na linha lateral-ventral direita e esquerda, em região medial do corpo em sentido crânio-caudal. O exame ultrassonográfico permitiu avaliar todo o ciclo reprodutivo nas serpentes. Nas avaliações ultrassonográficas das fêmeas pode-se definir as fases de desenvolvimento ovariano e ovidutal. Os folículos ovarianos durante a fase pré-vitelogênica foram visualizados como homogêneos e anecogênicos, em forma de "cacho de uva". Já na fase vitelogênica, os folículos estavam maiores e mais ecogênicos seguidos uns dos outros, como um "colar de pérolas". Quando não houve cópula, os folículos foram reabsorvidos dentro do ovário retornando a fase pré-vitelogênica. Na fase pós ovulatória foram visualizados três estágios bem definidos de desenvolvimento fetal dentro do oviduto: 1) logo após a ovulação (e fecundação), somente o vitelo foi visualizado; 2) o vitelo ocupava 60% e o feto 40% do ovo e 3) o feto estava formado e não havia vitelo. Nos machos, os testículos foram visualizados como uma imagem homogênea e hipoecogênica quando se encontravam em estágio reprodutivo. Quando não estavam reprodutivos não era possível visualizar a imagem do testículo devido ao seu tamanho. A avaliação ultrassonográfica do aparelho reprodutor em serpentes demonstrou ser uma técnica de diagnóstico segura, não invasiva e que permite o acompanhamento das principais fases reprodutivas

Análise comparativa da morfometria do casco de bovinos das raças Nelore, Curraleira e Pantaneira e de bubalinos e sua relação com a etiopatogenia das enfermidades digitais

O estudo morfométrico dos dígitos de bovinos e bubalinos pode colaborar para o entendimento da etiopatogenia das enfermidades podais. Este estudo objetivou descrever as características morfométricas dos dígitos de bovinos das raças Curraleira (Bos taurus), Pantaneira (Bos taurus), Nelore (Bos indicus) e de bubalinos (Bubalus bubalis) da raça Murrah e estabelecer possível relação entre tais medidas e a ocorrência de enfermidades digitais. Na pesquisa foram utilizados dez animais, saudáveis, de cada raça e espécie. Foram avaliados dois membros de cada animal, sendo um torácico e outro pélvico, totalizando 80 extremidades distais. As medidas morfométricas foram obtidas com auxílio de um paquímetro mecânico graduado e os ângulos das pinças conferidos por meio de transferidor metálico. Os principais parâmetros digitais avaliados foram o ângulo dorsal do casco (A), comprimento da parede dorsal (B), altura do talão (C), altura da pinça (D), comprimento do casco (E), comprimento diagonal do casco (F), largura do dígito lateral (G), largura do dígito medial (H), comprimento do dígito lateral (I) e comprimento do dígito medial (J). Para a comparação de médias dos resultados obtidos entre as raças foi utilizado o teste de Tukey (p<0,05). A análise multivariada para as representações gráficas das variáveis canônicas foi empregada para expressar a similaridade das medidas estudadas entre os grupos, no qual se utilizou o software R. Os resultados revelaram que os bubalinos apresentam as maiores medidas morfométricas para as variáveis B, C, D, E, F, G, H, I e J e apenas na variável A apresentaram medidas inferiores entre as diferentes raças de bovinos estudadas. Existe similaridade entre as três raças de bovinos estudadas em relação às variáveis, altura da pinça (D), largura do dígito lateral (G) e largura do dígito medial (H) as quais se distanciam dos valores encontrados para essas variáveis nos bubalinos, Concluiu que a morfometria digital pode influenciar na ocorrência de enfermidades digitais, mas não age como fator isolado, necessitando da interação com outros fatores estruturais, ambientais e de manejo para a manifestação dessas doenças.

Correlations between ANP concentrations in atria, plasma and cerebral structures and sodium chloride preference in Wistar rats

We determined whether ANP (atrial natriuretic peptide) concentrations, measured by radioimmunoassay, in the ANPergic cerebral regions involved in regulation of sodium intake and excretion and pituitary gland correlated with differences in sodium preference among 40 Wistar male rats (180-220 g). Sodium preference was measured as mean spontaneous ingestion of 1.5% NaCl solution during a test period of 12 days. The relevant tissues included the olfactory bulb (OB), the posterior and anterior lobes of the pituitary gland (PP and AP, respectively), the median eminence (ME), the medial basal hypothalamus (MBH), and the region anteroventral to the third ventricle (AV3V). We also measured ANP content in the right (RA) and left atrium (LA) and plasma. The concentrations of ANP in the OB and the AP were correlated with sodium ingestion during the preceding 24 h, since an increase of ANP in these structures was associated with a reduced ingestion and vice-versa (OB: r = -0.3649, P<0.05; AP: r = -0.3291, P<0.05). Moreover, the AP exhibited a correlation between ANP concentration and mean NaCl intake (r = -0.4165, P<0.05), but this was not the case for the OB (r = 0.2422). This suggests that differences in sodium preference among individual male rats can be related to variations of AP ANP level. Earlier studies indicated that the OB is involved in the control of NaCl ingestion. Our data suggest that the OB ANP level may play a role mainly in day-to-day variations of sodium ingestion in the individual rat

Interaction between NO and oxytocin: Influence on LHRH release

Nitric oxide synthase (NOS)-containing neurons have been localized in various parts of the CNS. These neurons occur in the hypothalamus, mostly in the paraventricular and supraoptic nuclei and their axons project to the neural lobe of the pituitary gland. We have found that nitric oxide (NO) controls luteinizing hormone-releasing hormone (LHRH) release from the hypothalamus acting as a signal transducer in norepinephrine (NE)-induced LHRH release. LHRH not only releases LH from the pituitary but also induces sexual behavior. On the other hand, it is known that oxytocin also stimulates mating behavior and there is some evidence that oxytocin can increase NE release. Therefore, it occurred to us that oxytocin may also stimulate LHRH release via NE and NO. To test this hypothesis, we incubated medial basal hypothalamic (MBH) explants from adult male rats in vitro. Following a preincubation period of 30 min, MBH fragments were incubated in Krebs-Ringer bicarbonate buffer in the presence of various concentrations of oxytocin. Oxytocin released LHRH at concentrations ranging from 0.1 nM to 1 µM with a maximal stimulatory effect (P<0.001) at 0.1 µM, but with no stimulatory effect at 10 µM. That these effects were mediated by NO was shown by the fact that incubation of the tissues with NG-monomethyl-L-arginine (NMMA), a competitive inhibitor of NOS, blocked the stimulatory effects. Furthermore, the release of LHRH by oxytocin was also blocked by prazocin, an a1-adrenergic receptor antagonist, indicating that NE mediated this effect. Oxytocin at the same concentrations also increased the activity of NOS (P<0.01) as measured by the conversion of [14C]arginine to citrulline, which is produced in equimolar amounts with NO by the action of NOS. The release of LHRH induced by oxytocin was also accompanied by a significant (P<0.02) increase in the release of prostaglandin E2 (PGE2), a mediator of LHRH release that is released by NO. On the other hand, incubation of neural lobes with various concentrations of sodium nitroprusside (NP) (300 or 600 µM), a releaser of NO, revealed that NO acts to suppress (P<0.01) the release of oxytocin. Therefore, our results indicate that oxytocin releases LHRH by stimulating NOS via NE, resulting in an increased release of NO, which increases PGE2 release that in turn induces LHRH release. Furthermore, the released NO can act back on oxytocinergic terminals to suppress the release of oxytocin in an ultrashort-loop negative feedback

Postangioplasty restenosis: a practical model in the porcine carotid artery

Transluminal coronary angioplasty is a routine therapeutic intervention in coronary heart disease. Despite the high rate of primary success, restenosis continues to be its major limitation. Porcine models have been considered to be the most adequate experimental models for studying restenosis. One limitation of porcine models is the need for radiological guidance and the expenses involved. The objective of the present study was to adapt an experimental model of angioplasty in the porcine carotid artery that does not require radiological equipment. Eight animals were used to develop the technique of balloon injury to the common carotid artery by dissection without radiological guidance. This technique was then employed in six other animals. Under anesthesia, the left common carotid artery was dissected and incised at the carotid sinus for insertion of an over-the-wire angioplasty balloon towards the aorta. Overstretch injury of the carotid artery was performed under direct visualization. After 30 days, the arteries were excised and pressure-fixated. Uninjured carotid arteries from 3 additional animals were used as controls. A decreased luminal area associated with intimal hyperplasia and medial reaction was observed in all injured arteries. Immunohistochemistry identified the intimal hyperplastic cells as smooth muscle cells. Computerized morphometry of the ballooned segments revealed the following mean areas: lumen 2.12 mm2 (± 1.09), intima 0.22 mm2 (± 0.08), media 3.47 mm2 (± 0.67), and adventitia 1.11 mm2 (± 0.34). Our experimental model of porcine carotid angioplasty without radiological guidance induced a vascular wall reaction and permitted the quantification of this response. This porcine model may facilitate the study of vascular injury and its response to pharmacological interventions

Faster gastric emptying of a liquid meal in rats after hypothalamic dorsomedial nucleus lesion

The effects of dorsomedial hypothalamic (DMH) nucleus lesion on body weight, plasma glucose levels, and the gastric emptying of a liquid meal were investigated in male Wistar rats (170-250 g). DMH lesions were produced stereotaxically by delivering a 2.0-mA current for 20 s through nichrome electrodes (0.3-mm tip exposure). In a second set of experiments, the DMH and the ventromedial hypothalamic (VMH) nucleus were lesioned with a 1.0-mA current for 10 s (0.1-mm tip exposure). The medial hypothalamus (MH) was also lesioned separately using a nichrome electrode (0.3-mm tip exposure) with a 2.0-mA current for 20 s. Gastric emptying was measured following the orogastric infusion of a liquid test meal consisting of physiological saline (0.9% NaCl, w/v) plus phenol red dye (6 mg/dl) as a marker. Plasma glucose levels were determined after an 18-h fast before the lesion and on the 7th and 15th postoperative day. Body weight was determined before lesioning and before sacrificing the rats. The DMH-lesioned rats showed a significantly faster (P<0.05) gastric emptying (24.7% gastric retention, N = 11) than control (33.0% gastric retention, N = 8) and sham-lesioned (33.5% gastric retention, N = 12) rats, with a transient hypoglycemia on the 7th postoperative day which returned to normal by the 15th postoperative day. In all cases, weight gain was slower among lesioned rats. Additional experiments using a smaller current to induce lesions confirmed that DMH-lesioned rats had a faster gastric emptying (25.1% gastric retention, N = 7) than control (33.4% gastric retention, N = 17) and VMH-lesioned (34.6% gastric retention, N = 7) rats. MH lesions resulted in an even slower gastric emptying (43.7% gastric retention, N = 7) than in the latter two groups. We conclude that although DMH lesions reduce weight gain, they do not produce consistent changes in plasma glucose levels. These lesions also promote faster gastric emptying of an inert liquid meal, thus suggesting a role for the DMH in the regulation of gastric motility

Antinociception induced by stimulating amygdaloid nuclei in rats: changes produced by systemically administered antagonists

The antinociceptive effects of stimulating the medial (ME) and central (CE) nuclei of the amygdala in rats were evaluated by the changes in the latency for the tail withdrawal reflex to noxious heating of the skin. A 30-s period of sine-wave stimulation of the ME or CE produced a significant and short increase in the duration of tail flick latency. A 15-s period of stimulation was ineffective. Repeated stimulation of these nuclei at 48-h intervals produced progressively smaller effects. The antinociception evoked from the ME was significantly reduced by the previous systemic administration of naloxone, methysergide, atropine, phenoxybenzamine, and propranolol, but not by mecamylamine, all given at the dose of 1.0 mg/kg. Previous systemic administration of naloxone, atropine, and propranolol, but not methysergide, phenoxybenzamine, or mecamylamine, was effective against the effects of stimulating the CE. We conclude that the antinociceptive effects of stimulating the ME involve at least opioid, serotonergic, adrenergic, and muscarinic cholinergic descending mechanisms. The effects of stimulating the CE involve at least opioid, ß-adrenergic, and muscarinic cholinergic descending mechanisms.

Peritoneal fluid modulates the sperm acrosomal exocytosis induced by N-acetylglucosaminyl neoglycoprotein

The effect of peritoneal fluid (PF) on the human sperm acrosome reaction (AR) was tested. Sperm was pre-incubated with PF and the AR was induced by calcium ionophore A23187 and a neoglycoprotein bearing N-acetylglycosamine residues (NGP). The AR induced by calcium ionophore was inhibited 40% by PF from controls (PFc) and 50% by PF from the endometriosis (PFe) group, but not by PF from infertile patients without endometriosis (PFi). No significant differences were found in the spontaneous AR. When the AR was induced by NGP, pre-incubation with PFc reduced (60%) the percentage of AR, while PFe and PFi caused no significant differences. The average rates of acrosome reactions obtained in control, NGP- and ionophore-treated sperm showed that NGP-induced exocytosis differed significantly between the PFc (11%) and PFe/PFi groups (17%), and the ionophore-induced AR was higher for PFi (33%) than PFc/PFe (25%). The incidence of the NGP-induced AR was reduced in the first hour of pre-incubation with PFc and remained nearly constant throughout 4 h of incubation. The present data indicate that PF possesses a protective factor which prevents premature AR.

Interaction between paraventricular nucleus and septal area in the control of physiological responses induced by angiotensin II

We determined the effects of losartan (40 nmol) and PD 123319 (40 nmol) (both non-peptides and selective antagonists of the AT1 and AT2 angiotensin receptors, respectively), and [Sar¹, Ala8] angiotensin II (ANG II) (40 nmol) (a non-selective peptide antagonist of angiotensin receptors) injected into the paraventricular nucleus (PVN) on the water and salt appetite, diuresis and natriuresis and mean arterial pressure (MAP) induced by administration of 10 nmol of ANG II into the medial septal area (MSA) of male Holtzman rats weighing 250-300 g. The volume of drug solution injected was 0.5 µl over a period of 10-15 s. The responses were measured over a period of 120 min. ANG II alone injected into the MSA induced an increase in all the above parameters (8.1 ± 1.2, 1.8 ± 0.3, and 17.1 ± 1.0 ml, 217 ± 25 µEq/120 min, and 24 ± 4 mmHg, respectively, N = 10-12) compared with vehicle-treated rats (1.4 ± 0.2, 0.6 ± 0.1, and 9.3 ± 0.5 ml, 47 ± 5 µEq/120 min, and 4.1 ± 0.8 mmHg, respectively, N = 10-14). Pretreatment with losartan and [Sar¹, Ala8] ANG II completely abolished the water and sodium intake, and the pressor increase (0.5 ± 0.2, 1.1 ± 0.2, 0.5 ± 0.2, and 0.8 ± 0.2 ml, and 1.2 ± 3.9, 31 ± 4.6 mmHg, respectively, N = 9-12), whereas losartan blunted the urinary and sodium excretion induced by ANG II (13.9 ± 1.0 ml and 187 ± 10 µEq/120 min, respectively, N = 9). Pretreatment with PD 123319 and [Sar¹, Ala8] ANG II blocked the urinary and sodium excretion (10.7 ± 0.8, 9.8 ± 0.7 ml, and 67 ± 13 and 57 ± 17 µEq/120 min, respectively, N = 9), whereas pretreatment with PD 123319 partially blocked the water and sodium intake, and the MAP induced by ANG II administration (2.3 ± 0.3, 1.1 ± 0.1 ml, and 12 ± 3 mmHg, respectively, N = 9-10). These results suggest the angiotensinergic effect of the MSA on the AT1 and AT2 receptors of the PVN in terms of water and sodium homeostasis and MAP modulation.

Nitrergic modulation of vasopressin, oxytocin and atrial natriuretic peptide secretion in response to sodium intake and hypertonic blood volume expansion

The central nervous system plays an important role in the control of renal sodium excretion. We present here a brief review of physiologic regulation of hydromineral balance and discuss recent results from our laboratory that focus on the participation of nitrergic, vasopressinergic, and oxytocinergic systems in the regulation of water and sodium excretion under different salt intake and hypertonic blood volume expansion (BVE) conditions. High sodium intake induced a significant increase in nitric oxide synthase (NOS) activity in the medial basal hypothalamus and neural lobe, while a low sodium diet decreased NOS activity in the neural lobe, suggesting that central NOS is involved in the control of sodium balance. An increase in plasma concentrations in vasopressin (AVP), oxytocin (OT), atrial natriuretic peptide (ANP), and nitrate after hypertonic BVE was also demonstrated. The central inhibition of NOS by L-NAME caused a decrease in plasma AVP and no change in plasma OT or ANP levels after BVE. These data indicate that the increase in AVP release after hypertonic BVE depends on nitric oxide production. In contrast, the pattern of OT secretion was similar to that of ANP secretion, supporting the view that OT is a neuromodulator of ANP secretion during hypertonic BVE. Thus, neurohypophyseal hormones and ANP are secreted under hypertonic BVE in order to correct the changes induced in blood volume and osmolality, and the secretion of AVP in this particular situation depends on NOS activity.

Effects of septal cholinergic lesion on rat exploratory behavior in an open-field

The medial septum participates in the modulation of exploratory behavior triggered by novelty. Also, selective lesions of the cholinergic component of the septohippocampal system alter the habituation of rats to an elevated plus-maze without modifying anxiety indices. We investigated the effects of the intraseptal injection of the cholinergic immunotoxin 192 IgG-saporin (SAP) on the behavior of rats in an open-field. Thirty-nine male Wistar rats (weight: 194-230 g) were divided into three groups, non-injected controls and rats injected with either saline (0.5 µl) or SAP (237.5 ng/0.5 µl). Twelve days after surgery, the animals were placed in a square open-field (120 cm) and allowed to freely explore for 5 min. After the test, the rats were killed by decapitation and the septum, hippocampus and frontal cortex were removed and assayed for acetylcholinesterase activity. SAP increased acetylcholinesterase activity in the septum, hippocampus and frontal cortex and decreased the total distance run (9.15 ± 1.51 m) in comparison to controls (13.49 ± 0.91 m). The time spent in the center and at the periphery was not altered by SAP but the distance run was reduced during the first and second minutes (2.43 ± 0.36 and 1.75 ± 0.34 m) compared to controls (4.18 ± 0.26 and 3.14 ± 0.25 m). SAP-treated rats showed decreased but persistent exploration throughout the session. These results suggest that septohippocampal cholinergic mechanisms contribute to at least two critical processes, one related to the motivation to explore new environments and the other to the acquisition and storage of spatial information (i.e., spatial memory).

Sulfated proteoglycans as modulators of neuronal migration and axonal decussation in the developing midbrain

Proteoglycans are abundant in the developing brain and there is much circumstantial evidence for their roles in directional neuronal movements such as cell body migration and axonal growth. We have developed an in vitro model of astrocyte cultures of the lateral and medial sectors of the embryonic mouse midbrain, that differ in their ability to support neuritic growth of young midbrain neurons, and we have searched for the role of interactive proteins and proteoglycans in this model. Neurite production in co-cultures reveals that, irrespective of the previous location of neurons in the midbrain, medial astrocytes exert an inhibitory or nonpermissive effect on neuritic growth that is correlated to a higher content of both heparan and chondroitin sulfates (HS and CS). Treatment of astrocytes with chondroitinase ABC revealed a growth-promoting effect of CS on lateral glia but treatment with exogenous CS-4 indicated a U-shaped dose-response curve for CS. In contrast, the growth-inhibitory action of medial astrocytes was reversed by exogenous CS-4. Treatment of astrocytes with heparitinase indicated that the growth-inhibitory action of medial astrocytes may depend heavily on HS by an as yet unknown mechanism. The results are discussed in terms of available knowledge on the binding of HS proteoglycans to interactive proteins, with emphasis on the importance of unraveling the physiological functions of glial glycoconjugates for a better understanding of neuron-glial interactions.

c-Fos expression induced by electroacupuncture at the Zusanli point in rats submitted to repeated immobilization

In laboratory animals, acupuncture needs to be performed on either anesthetized or, if unanesthetized, restrained subjects. Both procedures up-regulate c-Fos expression in several areas of the central nervous system, representing therefore a major pitfall for the assessment of c-Fos expression induced by electroacupuncture. Thus, in order to reduce the effect of acute restraint we used a protocol of repeated restraint for the assessment of the brain areas activated by electroacupuncture in adult male Wistar rats weighing 180-230 g. Repeated immobilization protocols (6 days, 1 h/day and 13 days, 2 h/day) were used to reduce the effect of acute immobilization stress on the c-Fos expression induced by electroacupuncture at the Zusanli point (EA36S). Animals submitted to immobilization alone or to electroacupuncture (100 Hz, 2-4 V, faradic wave) in a non-point region were compared to animals submitted to electroacupuncture at EA36S (4 animals/subgroup). c-Fos expression was measured in 41 brain areas by simple counting of cells and the results are reported as number of c-Fos-immunoreactive cells/10,000 µm². The protocols of repeated immobilization significantly reduced the immobilization-induced c-Fos expression in most of the brain areas analyzed (P < 0.05). Animals of the EA36S groups had significantly higher levels of c-Fos expression in the dorsal raphe nucleus, locus coeruleus, posterior hypothalamus and central medial nucleus of the thalamus. Furthermore, the repeated immobilization protocols intensified the differences between the effects of 36S and non-point stimulation in the dorsal raphe nucleus (P < 0.05). These data suggest that high levels of stress can interact with and mask the evaluation of specific effects of acupuncture in unanesthetized animals.