Brainstem oxytocinergic modulation of heart rate control in rats: effects of hypertension and exercise training

Oxytocinergic brainstem projections participate in the autonomic control of the circulation. We investigated the effects of hypertension and training on cardiovascular parameters after oxytocin (OT) receptor blockade within the nucleus tractus solitarii (NTS) and NTS OT and OT receptor expression. Male spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats were trained (55% of maximal exercise capacity) or kept sedentary for 3 months and chronically instrumented (NTS and arterial cannulae). Mean arterial blood pressure (MAP) and heart rate (HR) were measured at rest and during an acute bout of exercise after NTS pretreatment with vehicle or OT antagonist (20 pmol of OT antagonist (200 nl of vehicle)-1). Oxytocin and OT receptor were quantified (35S-oligonucleotide probes, in situ hybridization) in other groups of rats. The SHR exhibited high MAP and HR (P < 0.05). Exercise training improved treadmill performance and reduced basal HR (on average -11%) in both groups, but did not change basal MAP. Blockade of NTS OT receptor increased exercise tachycardia only in trained groups, with a larger effect on trained WKY rats (+31 +/- 9 versus +12 +/- 3 beats min-1 in the trained SHR). Hypertension specifically reduced NTS OT receptor mRNA density (-46% versus sedentary WKY rats, P < 0.05); training did not change OT receptor density, but significantly increased OT mRNA expression (+2.5-fold in trained WKY rats and +15% in trained SHR). Concurrent hypertension- and training-induced plastic (peptide/receptor changes) and functional adjustments (HR changes) of oxytocinergic control support both the elevated basal HR in the SHR group and the slowing of the heart rate (rest and exercise) observed in trained WKY rats and SHR.

Multiple Pathways Analysis of Brain Functional Networks from EEG Signals: An Application to Real Data

In the present study, we propose a theoretical graph procedure to investigate multiple pathways in brain functional networks. By taking into account all the possible paths consisting of h links between the nodes pairs of the network, we measured the global network redundancy R (h) as the number of parallel paths and the global network permeability P (h) as the probability to get connected. We used this procedure to investigate the structural and dynamical changes in the cortical networks estimated from a dataset of high-resolution EEG signals in a group of spinal cord injured (SCI) patients during the attempt of foot movement. In the light of a statistical contrast with a healthy population, the permeability index P (h) of the SCI networks increased significantly (P < 0.01) in the Theta frequency band (3-6 Hz) for distances h ranging from 2 to 4. On the contrary, no significant differences were found between the two populations for the redundancy index R (h) . The most significant changes in the brain functional network of SCI patients occurred mainly in the lower spectral contents. These changes were related to an improved propagation of communication between the closest cortical areas rather than to a different level of redundancy. This evidence strengthens the hypothesis of the need for a higher functional interaction among the closest ROIs as a mechanism to compensate the lack of feedback from the peripheral nerves to the sensomotor areas.

Increased SOD1 association with chromatin, DNA damage, p53 activation, and apoptosis in a cellular model of SOD1-linked ALS

Mutations in the gene encoding cytosolic Cu,Zn-superoxide dismutase (SOD1) have been linked to familial amyotrophic lateral sclerosis (FALS). However the molecular mechanisms of motor neuron death are multifactorial and remain unclear. Here we examined DNA damage;p53 activity and apoptosis in SH-SY5Y human neuroblastoma cells transfected to achieve low-level expression of either wild-type or mutant Gly(93) --> Ala (G93A) SOD1, typical of FALS. DNA damage was investigated by evaluating the levels of 8-oxo-7,8-dihydro-2`-deoxyguanosine (8-oxodGuo) and DNA strand breaks. Significantly higher levels of DNA damage, increased p53 activity, and a greater percentage of apoptotic cells were observed in SH-SY5Y cells transfected with G93A SOD1 when compared to cells overexpressing wild-type SOD1 and untransfected cells. Western blot, FACS, and confocal microscopy analysis demonstrated that G93A SOD1 is present in the nucleus in association with DNA. Nuclear G93A SOD1 has identical superoxide dismutase activity but displays increased peroxidase activity when compared to wild-type SOD1. These results indicate that the G93A mutant SOD1 association with DNA might induce DNA damage and trigger the apoptotic response by activating p53. This toxic activity of mutant SOD1 in the nucleus may play an important role in the complex mechanisms associated with motor neuron death observed in ALS pathogenesis. (C) 2010 Elsevier B.V. All rights reserved.

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.

Efeito da natação e do basquetebol em cadeira de rodas sobre o colesterol HDL: uma investigação em indivíduos com lesão medular

Despite the observation of an increase in life expectancy in individuals with Spinal cord injury (SCI), it is lower than that of the general population. Studies have shown that affected individuals have a sedentary lifestyle that reflects negatively on health and quality of life. Studies have demonstrated that HDL cholesterol (HDL-C) levels, a high-density lipoprotein and important predictor of cardiovascular disease, are lower in this population exposing these people to a greater incidence of heart disease from atherosclerotic process In the general population, exercise increases HDL-C serum levels, but this phenomenon is not very clear in people with spinal cord injury (SCI). The present study examined the effect of both swimming and wheelchair basketball in the lipid profile of eleven men and seven women with SCI. The subjects included in regular exercise programs showed increases in HDL-C levels and decreases in CT/HDL-C and LDL-C/HDL-C ratios. We found better results mainly in men with lower levels of SCI and in those that sustained exercise intensities above 60% of the heart rate reserve. The duration of training sessions can be an essential factor in these results. The results suggest that both the exercise prescription and the personal characteristics of people with SCI influence changes in the lipid profile mediated through exercise. The elaboration of this work is an attempt to clarify uncertainties about health and the longevity of people with SCI generated in discussion of all members of the interdisciplinary rehabilitation team, especially the physiotherapists, nutritionists, nurses and physicians that contributed considerably in all phases of the research

Transplacental transmission of Toxoplasma gondii in reinfected pregnant female canines

Twelve pregnant female canines, naturally infected with Toxoplasma gondii, were reinfected with T. gondii: three (GI) received tachyzoites subcutaneously (1.0 x 107), three (GII) were orally inoculated with oocysts (1.5 x 104), and six (GIII) were kept as a nonreinfected control group. All the reinfected female canines (GI and GII) miscarried or presented fetal death, while only one GIII female presented a stillborn in a litter of four pups (P < 0.01). Fever, lymphoadenopathy, miscarriage, and fetal death were the main clinical alterations observed. The highest serological titers detected through the indirect fluorescence antibody test (IFAT) were 1,024 (GI) and 4,096 (GII). In group III, the titers ranged between 64 and 256. By bioassays in mice, T. gondii was isolated in 17 organs of the reinfected adult canines, in 11 of the control group, and in 20 of the neonates. Positive immunostaining of cysts and/or tachyzoites were observed in 26 canine tissues (14 from GI and GII and ten from GIII). The agent was detected by immunohistochemistry in the encephalon of a neonate and in the spinal cord of a stillborn, thus, confirming that T. gondii infected canine fetuses, provoking miscarriages, even in bitches that presented primoinfection.

Biochemical, histopathological and clinical evaluation of delayed effects caused by methamidophos isoforms and TOCP in hens: Ameliorative effects using control of calcium homeostasis

This work evaluated the potential of the isoforms of methamidophos to cause organophosphorus-induced delayed neuropathy (OPIDN) in hens. In addition to inhibition of neuropathy target esterase (NTE) and acetylcholinesterase (AChE), calpain activation, spinal cord lesions and clinical signs were assessed. The isoforms (+)-, (+/-)- and (-)-methamidophos were administered at 50 mg/kg orally; tri-ortho-cresyl phosphate (TOCP) was administered (500 mg/kg, po) as positive control for delayed neuropathy. The TOCP hens showed greater than 80% and approximately 20% inhibition of NTE and AChE in hen brain, respectively. Among the isoforms of methamidophos, only the (+)-methamidophos was capable of inhibiting NTE activity (approximately 60%) with statistically significant difference compared to the control group. Calpain activity in brain increased by 40% in TOCP hens compared to the control group when measured 24h after dosing and remained high (18% over control) 21 days after dosing. Hens that received (+)-methamidophos had calpain activity 12% greater than controls. The histopathological findings and clinical signs corroborated the biochemical results that indicated the potential of the (+)-methamidophos to be the isoform responsible for OPIDN induction. Protection against OPIDN was examined using a treatment of 2 doses of nimodipine (1 mg/kg, i.m.) and one dose of calcium gluconate (5 mg/kg, iv.). The treatment decreased the effect of OPIDN-inducing TOCP and (+)-methamidophos on calpain activity, spinal cord lesions and clinical signs. (C) 2012 Elsevier B.V. All rights reserved.

Plasticidade induzida por treinamento locomotor na medula espinal intacta em ratos: correlatos morfológicos

The locomotion is one of the most important capabilities developed by the animals, whose improvement is dependent on several neural centers, including the spinal cord. This activity promotes a lot of spinal modifications that enable it to adapt and improve their connections. This study aimed to observe the morphological changes occurring in the spinal cord after locomotor training in intact rats. For that we used male Wistar rats, which were submitted to locomotor training in wheel activity in protocols 1, 3 and 7 days (30min/day), and the results were compared to a control group not subjected to exercise. Coronal sections of 40 μm of the lumbosacral spinal cord were subjected to immunohistochemical techniques anti-Egr1, anti-NMDA and anti-SP, to characterize the spinal plasticity related to these substances. Egr1-immunoreactive cells were increased in all laminas, essentially in those more intensely activated by locomotion, laminas IV-X levels L4-S3. All observed sections expressed NMDA-immunoreactivity. Analysis of SP in the spinal dorsal horn resulted no significant variations of this neuropeptide related to locomotion. The results suggest that locomotor training provides synaptic plasticity similar to LTP in all laminas of the lumbosacral spinal cord, in different intensities. However, the SP appears do not participate of this process in the spinal dorsal horn. This work will contribute for consolidating and characterization of synaptic plasticity in the spinal cord

The Neuraxial Effects of Intraspinal Amitriptyline at Low Concentrations

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos da administração subaracnóidea de grandes volumes de lidocaína a 2% e ropivacaína a 1% sobre a medula espinhal e as meninges: estudo experimental em cães

JUSTIFICATIVA E OBJETIVOS: A injeção de grandes volumes de anestésico local no espaço subaracnóideo, após punção dural acidental, é complicação da anestesia peridural. O objetivo desta pesquisa foi investigar as possíveis alterações clínicas e histológicas desencadeadas por grandes volumes de lidocaína a 2% e ropivacaína a 1%, simulando injeção subaracnóidea acidental, em cães. MÉTODO: Vinte e um cães foram distribuídos aleatoriamente em 3 grupos, que receberam por via subaracnóidea: G1 - cloreto de sódio a 0,9%; G2 - lidocaína a 2% e G3 - ropivacaína a 1%. A punção subaracnóidea foi realizada no espaço intervertebral L6-L7. O volume de anestésico local administrado foi de 1 ml para cada 10 cm de distância entre a protuberância occipital e o espaço lombossacral (5 - 6,6 ml). Após 72 horas de observação clínica os animais foram sacrificados e foi removida a porção lombossacral da medula para exame histológico, por microscopia óptica. RESULTADOS: Nenhum animal do G1 apresentou alterações clínicas ou histológicas da medula espinhal. Foram observados dois casos de necrose do tecido nervoso em G2, porém mudanças clínicas, em somente um desses cães e em outros dois animais que não apresentaram alterações histológicas. Foi encontrada necrose focal do tecido nervoso medular em um animal de G3. Todos os animais de G3 permaneceram clinicamente normais. CONCLUSÕES: Conclui-se que grandes volumes de lidocaína a 2% determinaram alterações clínicas e histológicas mais intensas que os de ropivacaína a 1%.

Clinical and Histological Effects of the Intrathecal Administration of Methylprednisolone in Dogs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Respiratory muscular strength decrease in children with mylomeningocele

Study Design. Case-control study.Objective. To evaluate respiratory muscle force in children with myelomeningocele. Summary of Background Data. Myelomeningocele is a common spinal cord malformation with limitations linked to central nervous system lesions and abnormalities in respiratory movements. Despite this, little attention has been given to evaluating respiratory muscle force in these patients.Methods. Children with myelomeningocele aged between 4 and 14 years ( myelomeningocele group; MG, n = 20) were studied and compared with healthy children ( control group; CG, n = 20) matched for age and gender. Respiratory muscular force was evaluated by maximum inspiratory ( Pimax) and expiratory ( Pemax) pressures.Results. Groups were similar for age [ CG = 8 ( 6 - 13) = MG = 8 ( 4 - 14), P > 0.05]; gender, and body mass index [ CG = 17.4 ( 14.1 - 24.7) x MG = 19.2 ( 12.6 - 31.9), P > 0.05]. The lumbosacral region was predominantly affected ( 45%). Maximum respiratory pressures were significantly higher in CG than MG ( Pimax = CG: similar to 83 +/- 21.75 > MG: -54.1 +/- 23.66; P < 0.001 and Pemax = CG: + 87.4 +/- 26.28 > MG: + 64.6 +/- 26.97; P = 0.01). Patients with upper spinal lesion ( UL) had lower maximum respiratory pressure values than those with lower spinal lesion ( LL), [Pimax ( UL = - 38.33 +/- 11.20 cm H2O x LL = - 60.85 +/- 24.62 cm H2O), P < 0.041 and Pemax ( UL = + 48 +/- 20.82 cm H2O x LL + 71.71 +/- 26.73 cm H2O), P = 0.067]).Conclusion. Children with myelomeningocele at the ages studied presented reduced respiratory muscle force with more compromise in upper spinal lesion.

The heminested RT-PCR for the study of rabies virus pathogenesis

The aim of the present trial was to evaluate the heminested RT-PCR for the study of rabies virus distribution in mice inoculated experimentally. Inoculation was by the intramuscular route in 150 mice, using the dog street rabies virus. Groups of five animals were killed at different times. Fragments of different organs were collected and the material was tested by Fluorescent Antibody Test (FAT) and heminested RT-PCR (hn RT-PCR). Positive results were obtained beginning on the 10th day after inoculation in the brain, spinal cord, salivary gland, limbs, lungs, liver, spleen, urinary bladder, tongue and right kidney. Hn RT-PCR was shown to be more efficient for the study of rabies virus distribution in different tissues and organs. (C) 2004 Elsevier B.V.. All rights reserved.

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

AV3V lesions reduce the pressor response to L-glutamate into the RVLM

Neurons from the rostral ventrolateral medulla (RVLM) directly activate sympathetic preganglionic neurons in the spinal cord. Hypertensive responses and sympathetic activation produced by different stimuli are strongly affected by lesions of the preoptic periventricular tissue surrounding the anteroventral third ventricle (AV3V region). Therefore, in the present study, we investigated the effects of acute (1 day) and chronic (IS days) electrolytic lesions of the AV3V region on the pressor responses produced by injections of the excitatory amino acid L-glutamate into the RVLM of unanesthetized rats. Male Holtzman rats with sham or electrolytic AV3V lesions and a stainless steel cannula. implanted into the RVLM were used. The pressor responses produced by injections of L-glutamate (1, 5 and 10 nmol/100 nl) into the RVLM were reduced 1 day (9 +/- 4, 39 +/- 6 and 37 +/- 4 mm Hg, respectively) and 15 days after AV3V lesions (13 +/- 6, 39 +/- 4 and 43 +/- 4 mm Hg, respectively, vs. sham lesions: 29 +/- 3, 50 +/- 2 and 58 +/- 3 mm Hg, respectively). Injections of L-glutamate into the RVLM in sham or AV3V-lesioned rats produced no significant change in the heart rate (HR). Baroreflex bradycardia and tachycardia produced by iv phenylephrine or sodium nitroprusside, respectively, and the pressor and bradycardic responses to chemoreflex activation with iv potassium cyanide were not modified by AV3V lesions. The results suggest that signals from the AV3V region are important for sympathetic activation induced by L-glutamate into the RVLM. (c) 2006 Elsevier B.V. All rights reserved.