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AIMS: To identify the bacteria nodulating Sulla spinosissima growing profusely in a lead and zinc mine tailings in Eastern Morocco. METHODS AND RESULTS: In all, 32 rhizobial cultures, isolated from root nodules of S. spinosissima growing in soils of the mining site, were tolerant to different heavy metals. The ERIC-polymerase chain reaction (PCR) fingerprinting analysis clustered the isolates into seven different groups, and the analysis of the 16S rRNA sequences of four selected representative strains, showed they were related to different species of the genus Mesorhizobium. The atpD, glnII and recA housekeeping genes analysis confirmed the affiliation of the four representative strains to Mesorhizobium camelthorni CCNWXJ40-4T , with similarity percentages varying from 96·30 to 98·30%. The sequences of the nifH gene had 97·33-97·78% similarities with that of M. camelthorni CCNWXJ40-4T ; however, the nodC phylogeny of the four strains diverged from the type and other reference strains of M. camelthorni and formed a separated cluster. The four strains nodulate also Astragalus gombiformis and A. armatus but did not nodulate A. boeticus, Vachellia gummifera, Prosopis chilensis, Cicer arietinum, Lens culinaris, Medicago truncatula, Lupinus luteus or Phaseolus vulgaris. CONCLUSIONS: Based on similarities of the nodC symbiotic gene and differences in the host range, the strains isolated from S. spinosissima growing in soils of the Sidi Boubker mining site may form a different symbiovar within Mesorhizobium for which the name aridi is proposed. SIGNIFICANCE AND IMPACT OF THE STUDY: In this work, we show that strains of M. camelthorni species nodulating S. spinosissima in the arid area of Eastern Morocco constitute a distinct phylogenetic clade of nodulation genes; we named symbiovar aridi, which encompasses also mesorhizobia from other Mediterranean desert legumes.
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Fabaceae/microbiologia , Chumbo/metabolismo , Mesorhizobium/fisiologia , Mineração , Simbiose , Proteínas de Bactérias/genética , Genes Essenciais/genética , Especificidade de Hospedeiro , Mesorhizobium/classificação , Marrocos , Filogenia , Nodulação/genética , RNA Ribossômico 16S/genética , Nódulos Radiculares de Plantas/microbiologia , Microbiologia do Solo , Simbiose/genéticaRESUMO
Purpose: The bioluminescence reporter PER2::Luciferase (PER2::Luc) provides a powerful tool to study the regulation of biological clocks in explant tissues, including the retinal clock. However, the establishment of a standardized procedure to replicate experimental conditions and to enable meaningful comparisons between findings from different studies is still lacking. In addition, different parameters may affect the retinal circadian bioluminescence signal and its dynamic in in vitro assays. In the present study, we first evaluated the effect of sex and age on the main parameters of the mouse retinal clock. We then examined the impact of medium change on PER2::Luc rhythm and compared two light stimulation protocols of the retinal clock. Methods: In a first set of experiments, retinal explants from both male and female Per2Luc mice of different ages (1 to 8 months) are cultured and the period, phase, amplitude, and rhythmic power of PER2::Luc oscillations are analyzed. In a second set of experiments, we quantified the effect of a medium change done after 4, 6, 8, 9, or 10 days of culture on the phase and period of retinal explants. Finally, we compared the phase shift and the period change resulting from two methods of light stimulations of retinal explants: the first involved the transfer of the cultured tissues from the Lumicycle into a light stimulation chamber, while the second used a light delivery apparatus embedded in the Lumicycle. Results: We do not observe any sex-dependent effects on the amplitude, period, phase, and rhythmic power of the in vitro retinal PER2::Luc oscillations in animals aged of 2 to 3 months. The most remarkable effect of age is on the amplitude of PER2::Luc oscillations that significantly decrease from 1 to 4-5 months, whereas the endogenous period and rhythmic power increase slightly until 2 to 3 months and then do not change until 8 months. The phase is not affected by age. We then show that a medium change occurring after 4 days of culture does not alter the phase of PER2::Luc rhythm by comparison with day 0, whereas a medium change done after 6, 8, 9, or 10 days in culture advances the phase and lengthens the period. Finally, we observe that the physical displacement of the culture dishes containing retinal explants, even in complete darkness, induces a strong phase shift of PER2::Luc oscillations. Conclusions: Our work shows that the retina cultures are particularly sensitive to some aspects of the culture procedure, and it provides an accurate standard protocol to avoid biases due to artifactually induced phase shifts resulting from the medium change or physical displacement.
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Técnicas de Cultura de Células/métodos , Relógios Circadianos , Genes Reporter/genética , Medições Luminescentes/métodos , Proteínas Circadianas Period/metabolismo , Retina/metabolismo , Envelhecimento/fisiologia , Envelhecimento/efeitos da radiação , Animais , Relógios Biológicos/fisiologia , Relógios Biológicos/efeitos da radiação , Células Cultivadas , Relógios Circadianos/efeitos da radiação , Ritmo Circadiano/fisiologia , Ritmo Circadiano/efeitos da radiação , Feminino , Luz , Luciferases/metabolismo , Medições Luminescentes/instrumentação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas Circadianas Period/genética , Retina/efeitos da radiaçãoRESUMO
AIM: The best form of prophylactic management of a decompressed sigmoid volvulus (SV) is controversial especially in the elderly. We have studied our experience with this condition to assess the short- and long-term results of SV management. METHOD: All patients treated for SV in our department between 2003 and 2013 were retrospectively included. Emergency decompression was attempted in all patients in whom there was no sign of peritonitis. Planned surgical resection was the procedure of choice in young patients. Percutaneous endoscopic colopexy (PEC) was used in high surgical risk patients. RESULTS: There were 65 patients (45 males) of median age 71.5 (24-99) years. Non-surgical reduction was performed in 62 with a success rate of 95% (59/62). Recurrence after initial decompression was 67% at a median follow-up of 5 (1-14) years. A prophylactic surgical resection was performed with primary anastomosis in 33 patients. There were no deaths and the major morbidity rate was 6%. At a mean follow-up of 62 months, only 1 (3%) patient had had a recurrence (at 130 months). PEC was performed in six patients of median age 90 (84-99) years and with a median American Society of Anesthesiologists score of 4. Complications included local site infection (n = 2), pain (n = 1) and abdominal wall bleeding (n = 1). After a median follow-up of 2 (1-4) years, three patients died from medical causes and one recurrence occurred 13 months after removal of the PEC tube. CONCLUSION: Prophylactic treatment after initial decompression of SV results in a low rate of recurrence. Planned sigmoid resection is safe and effective. In frail elderly patients, PEC is satisfactory.
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Colectomia/métodos , Colo Sigmoide/cirurgia , Descompressão Cirúrgica/métodos , Volvo Intestinal/cirurgia , Doença Aguda , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Anastomose Cirúrgica , Estudos de Coortes , Colo Sigmoide/fisiopatologia , Colonoscopia/métodos , Tratamento de Emergência , Feminino , Seguimentos , Humanos , Volvo Intestinal/diagnóstico , Volvo Intestinal/mortalidade , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Estudos Retrospectivos , Medição de Risco , Fatores Sexuais , Estatísticas não Paramétricas , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: This study aimed to explore the occurrence and determinants of poor response to short-term pre-operative erythropoietin treatment and the effect of such poor response on transfusion in total hip arthroplasty patients. METHODS: We studied total hip arthroplasty patients who received erythropoietin before surgery. The primary outcome was the pre-operative increase in haemoglobin (delta haemoglobin) as response to erythropoietin therapy. Additionally, patients were classified in tertiles based on this delta haemoglobin: poor responders (cases), responders and good responders (controls) to erythropoietin. Patient characteristics, comedication and co-morbidity were collected as potential determinants of erythropoietin response. Regression techniques were used to estimate the strength of the associations and to assess the effect of poor response on transfusion requirement. RESULTS: A total of 379 patients receiving erythropoietin were eligible to enter the study. Mean delta haemoglobin was 19.3 g/l (standard deviation 9.4). Factors significantly associated with delta haemoglobin were the use of angiotensin II antagonists [-3.1 g/l; 95% confidence interval (CI) -5.7 to -0.6] and vitamin K antagonists (-6.9 g/l; 95% CI -10.0 to -0.2), together with body mass index (BMI) (-0.3 g/l per unit>; 95% CI -0.5 to -0.2). The additional case-control analysis yielded comparable results. Poor response to erythropoietin was associated with an increased transfusion risk (odds ratio 4.6, 95% CI 2.0-11). CONCLUSION: Use of angiotensin II receptor antagonists and vitamin K antagonists, and having a high BMI were determinants of poor response to short-term pre-operative erythropoietin treatment in total hip arthroplasty patients. Poor responders had a higher risk for perioperative blood transfusion.
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Anemia/tratamento farmacológico , Eritropoetina/uso terapêutico , Cuidados Pré-Operatórios , Idoso , Anemia/sangue , Artroplastia de Quadril , Transfusão de Sangue , Comorbidade , Interpretação Estatística de Dados , Feminino , Hemoglobinas/análise , Humanos , Masculino , Proteínas Recombinantes/uso terapêutico , Falha de Tratamento , Resultado do TratamentoRESUMO
Many experimental approaches require housing rodents in individual cages, including in epilepsy research. However, rats and mice are social animals; and individual housing constitutes a stressful situation. The goal of the present study was to determine the effects of individual housing as compared to conditions maintaining social contact on stress markers and epilepsy. Control male mice socially housed during pretest and then transferred to individual cages for six weeks displayed anhedonia, increased anxiety and biological markers of stress as compared to pretest values or mice kept socially housed during six weeks. Pilocarpine (pilo)-treated mice housed together showed increased levels of anhedonia, anxiety and stress markers as well as decreased cognitive performance as compared to the control group. The differences were more significant in pilo-treated mice housed individually. Anxiety correlated linearly with cognitive performance and stress markers independently of the experimental conditions. In the male rat pilo model, seizures were sixteen times more frequent in singly housed animals as compared to animals kept in pairs. Daily interactions with an experimenter in otherwise singly housed animals was sufficient to produce results identical to those found in animals kept in pairs. We propose that social isolation produces a severe phenotype in terms of stress and seizure frequency as compared to animals maintaining social contact (at least in these two models), a factor that needs to be taken into account for data interpretation, in particular for preclinical studies.
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Epilepsia/fisiopatologia , Abrigo para Animais , Convulsões/fisiopatologia , Isolamento Social , Estresse Psicológico/fisiopatologia , Anedonia/fisiologia , Animais , Ansiedade/complicações , Ansiedade/fisiopatologia , Modelos Animais de Doenças , Epilepsia/induzido quimicamente , Epilepsia/complicações , Masculino , Pilocarpina/administração & dosagem , Ratos Wistar , Convulsões/induzido quimicamente , Convulsões/complicações , Estresse Psicológico/complicaçõesRESUMO
PURPOSE: To evaluate the feasibility of day surgery anterior cruciate ligament reconstruction (ACLR) in children. Our hypothesis was that proper patient selection, adequate information and a streamlined clinical pathway would allow for a safe and satisfactory procedure. METHODS: A total of 20 patients, seven girls and 13 boys, who underwent outpatient ACLR at mean age of 14 years (8 to 16) were included in this single institution prospective case series. Surgery took place under simple general anesthesia while analgesia protocol combined local anesthesia, oral analgesics for the postoperative period and compressive cryotherapy. The main failure criterion was the inability to return home the day of surgery or the need for rehospitalization during the first week after discharge. Secondary evaluation parameters were the rate of postoperative complications, postoperative pain, quality of life (QOL) and patients' satisfaction outcome questionnaire (PSOQ) at postoperative day 1, 7 and 31. RESULTS: Of the 20 patients included, only one failed the ambulatory mode. The mean PSOQ score was, respectively for children and parents, 91.4% and 90.7% at day 7 and 94.6% and 95.7% at one month. Postoperative QOL at day 7 was very satisfying for all patients. Two minor early complications were reported. Both of the late complications, which included one partial wound dehiscence and one persistent knee swelling, resolved spontaneously. CONCLUSION: This study indicates that, in a targeted and well-informed population, day surgery for children ACLR yields high level of patient and family satisfaction in a safely manner. LEVEL OF EVIDENCE: IV, prospective case series.
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OBJECTIVE: This study aimed to investigate pregnancy outcomes in Somali-born women compared with those women born in each of the six receiving countries: Australia, Belgium, Canada, Finland, Norway and Sweden. DESIGN: Meta-analyses of routinely collected data on confinements and births. SETTING: National or regional perinatal datasets spanning 3-6 years between 1997 and 2004 from six countries. SAMPLE: A total of 10 431 Somali-born women and 2 168 891 receiving country-born women. METHODS: Meta-analyses to compare outcomes for Somali-born and receiving country-born women across the six countries. MAIN OUTCOME MEASURES: Events of labour (induction, epidural use and proportion of women using no analgesia), mode of birth (spontaneous vaginal birth, operative vaginal birth and caesarean section) and infant outcomes (preterm birth, birthweight, Apgar at 5 minutes, stillbirths and neonatal deaths). RESULTS: Compared with receiving country-born women, Somali-born women were less likely to give birth preterm (pooled OR 0.72, 95% CI 0.64-0.81) or to have infants of low birthweight (pooled OR 0.89, 95% CI 0.82-0.98), but there was an excess of caesarean sections, particularly in first births (pooled OR 1.41, 95% CI 1.25-1.59) and an excess of stillbirths (pooled OR 1.86, 95% CI 1.38-2.51). CONCLUSIONS: This analysis has identified a number of disparities in outcomes between Somali-born women and their receiving country counterparts. The disparities are not readily explained and they raise concerns about the provision of maternity care for Somali women postmigration. Review of maternity care practices followed by implementation and careful evaluation of strategies to improve both care and outcomes for Somali women is needed.
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Parto Obstétrico/estatística & dados numéricos , Emigração e Imigração/estatística & dados numéricos , Resultado da Gravidez/etnologia , Adulto , Austrália/epidemiologia , Canadá/epidemiologia , Europa (Continente)/epidemiologia , Feminino , Humanos , Mortalidade Infantil , Recém-Nascido , Idade Materna , Gravidez , Nascimento Prematuro/etnologia , Fatores de Risco , Somália/etnologia , Adulto JovemRESUMO
OBJECTIVE: Local recurrence of pelvic cancer is a therapeutic challenge. The purpose of the study was to evaluate radiofrequency ablation (RFA, intra-operative or CT-guided) for the treatment of pelvic recurrence in patients not eligible for curative surgical resection. METHOD: Charts of all patients treated for pelvic recurrence by RFA between March 2004 and March 2005 were reviewed. RESULTS: Eight patients (two females) had RFA for inoperable local recurrence [rectal adenocarcinoma (six) and sarcoma (two)]. Surgical resection of the primary tumour had been performed at a median age of 50.2 (36.7-61.6) years. Recurrence occurred after a median of 49.5 (11.7-63.5) months. The mean size of the recurrence was 33.4 (20-45) mm. RFA was given on a median number of two occasions (1-3). Complications occurred in six patients including minor pain [pelvic (six); sciatic nerve irritation (four)]; ureteric obstruction requiring stenting (two) and colo-vesical fistula (one), requiring ileal urinary diversion. After a median follow-up of 18.2 months (11-32), six patients were still alive. Patients, who had experienced pain prior to RFA were pain-free. Five patients showed evidence of further tumour growth but were asymptomatic. CONCLUSION: Radiofrequency ablation is a feasible therapeutic option for recurrent pelvic cancer. It allows good symptom control in patients with pain but morbidity is high.
Assuntos
Ablação por Cateter/métodos , Recidiva Local de Neoplasia/mortalidade , Recidiva Local de Neoplasia/cirurgia , Cuidados Paliativos/métodos , Neoplasias Pélvicas/mortalidade , Neoplasias Pélvicas/cirurgia , Adenocarcinoma/mortalidade , Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Adulto , Ablação por Cateter/efeitos adversos , Feminino , Seguimentos , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Medição da Dor , Neoplasias Pélvicas/patologia , Neoplasias Retais/mortalidade , Neoplasias Retais/patologia , Neoplasias Retais/cirurgia , Medição de Risco , Estudos de Amostragem , Sarcoma/mortalidade , Sarcoma/patologia , Sarcoma/cirurgia , Análise de Sobrevida , Resultado do TratamentoRESUMO
For powder catalyst characterization, Fourier Transform Infrared (FTIR), Raman, and X-Ray Fluorescence (XRF) spectrometers and X-Ray Diffraction (XRD) are available in high-throughput (HT) configurations, for example at the REALCAT platform to sequentially analyse multiple sets of samples. To remove the bottleneck resulting from the use of different sample holders for each equipment, a unique multi-well plate was developed. This paper details the design of such a plate including the selection of the fabrication material and the plate dimensioning based on the study of the 4 different physical interactions between matter and electromagnetic radiations for the aforementioned techniques. This new plate consists of a holder for removable wells enabling the avoidance of cross-contamination between samples. Raman, a focusing technique, has no strict constraint on the plate design. The number of wells, their geometry, spacing and dimensions were adjusted to deal with the constraints of IR optics. The well depth was set according to the XRF maximum penetration depth in the sample. The well diameter was optimized in order to obtain from the X-ray spot size the maximum achievable intensity. Poly-methyl-methacrylate (PMMA) was chosen as the material for the new plate due to its amorphous structure (no peak in XRD analysis) and ease with which it can be cut by a laser. Finally, the flatness of the multi-well plate was validated on the most challenging instrument: XRD. This new plate allows fast sample filling/preparation, requires small quantities of catalyst (50 to 80 mg) in each well and is compatible and convenient for HT experimentation.
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The number of axons in the optic nerve of the ovoviviparous reptile Vipera aspis was estimated from electron micrographs taken during the first 5 weeks of postnatal life. One to two days after birth, the optic nerve contains about 170,000 fibres, of which about 9% are myelinated. At the end of the fifth postnatal week, the number of optic fibres has fallen to about 100,000, of which about 42% are myelinated. This fibre loss continues after the fifth postnatal week, since in the adult viper the nerve contains about 60,000 fibres, of which 85% are myelinated; overall, about 65% of the optic nerve fibres present at birth disappear before the number of axons stabilises at the adult level. This study shows, for the first time, that the mode of development of the visual axons of reptiles is not that of anamniote vertebrates but similar to that of birds and mammals.
Assuntos
Nervo Óptico/crescimento & desenvolvimento , Nervo Óptico/ultraestrutura , Viperidae/crescimento & desenvolvimento , Animais , Animais Recém-Nascidos , Contagem de Células , Feminino , Masculino , Necrose/patologia , Fibras Nervosas/patologia , Fibras Nervosas/ultraestrutura , Fibras Nervosas Mielinizadas/patologia , Fibras Nervosas Mielinizadas/ultraestrutura , Neuroglia/ultraestrutura , Nervo Óptico/citologia , Viviparidade não MamíferaRESUMO
The aim of the present study was to analyze the neurochemical properties of the centrifugal visual system (CVS) of the quail using an immunohistochemical approach by testing 16 neuropeptides (angiotensin: ANG, bradykinin: BK, cholecystokinin, dynorphin, L and M-enkephalin, beta-endorphin: beta-END, galanin, alpha-neoendorphin, neurokinin A, neuropeptide Y (NPY), ocytocin, somatostatin, substance P, vasopressin, vasoactive intestinal polypeptide) and three neurotransmitters or their synthetic enzymes (choline acetyltransferase: ChAT, tyrosine hydroxylase: TH, serotonin: 5-HT and nitric oxide synthase: NOS, including the histochemical nicotinamide adenine dinucleotide phosphate diaphorase technique). For each substance, the somatic and afferent fiber and terminal labeling was analyzed within the nucleus isthmo-opticus (NIO) and the ectopic area (EA) and compared with that of retinopetal cell bodies labeled retrogradely with RITC following its intraocular injection (double-labeling procedure). The results showed that none of the centrifugal neurons were reactive to any of the substances tested. In contrast, all with the exception of ANG, BK and beta-END, labeled fibers and terminals within the EA and only four (ChAT, 5-HT, NPY and NOS) within the NIO. Possible sources of these immunoreactive fibers terminating in the NIO and EA were investigated by mapping the somatic immunolabeling of the different substances within brainstem regions previously shown by Miceli and other authors to project upon the centrifugal neurons. The data suggests that, besides the rapid retino-tecto-NIO-retinal loop, which facilitates the transfer of meaningful or more relevant information within particular portions of the visual field, the multiple afferent input which stems from various brainstem regions utilizes a wide range of neuroactive substances. Some of these afferent projections upon the centrifugal neurons appear to belong to nonspecific systems which might play a role in modulating the excitability of centrifugal neurons as a function of arousal.
Assuntos
Coturnix/fisiologia , Neurotransmissores/fisiologia , Visão Ocular/fisiologia , Animais , Avidina , Axônios/fisiologia , Biotina , Técnica Direta de Fluorescência para Anticorpo , Imuno-Histoquímica , Fibras Nervosas/metabolismo , Fibras Nervosas/fisiologia , Neuropeptídeos/metabolismo , Neurotransmissores/metabolismo , Vias Visuais/fisiologiaRESUMO
Expression of c-fos in L(5)-S(1) spinal segments in response to mechanical vaginocervical stimulation was investigated in both cycling and ovariectomized females. The aim of this paper was to verify the influence of estrous cycle on females genital tract sensitivity using immunodetection of a neural activity endogenous marker. The results indicate that lumbosacral spinal Fos-labeling was highly increased in vaginocervical stimulated rats relative to control, and labeled neurons were present more intensively in the dorsal horn in comparison to other spinal areas. Significant differences in Fos-labeling were observed according to the estrous cycle stage at which the stimulation was applied. In estrous females, the response was greater than that obtained at diestrous and much greater than the response of proestrous females. The spinal Fos-labeling of ovariectomized females is equivalent to that of diestrous females. These results give evidence that the vaginocervical induced expression of c-fos is modulated by cyclic changes in circulating sex hormones, whereas results observed in ovariectomized females indicate the likely involvement of other mechanisms independent of ovarian hormones.
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Colo do Útero/fisiologia , Estro/fisiologia , Regulação da Expressão Gênica/fisiologia , Genes fos , Medula Espinal/fisiologia , Vagina/fisiologia , Animais , Feminino , Imuno-Histoquímica , Neurônios/citologia , Neurônios/fisiologia , Ovariectomia , Pressão , Proteínas Proto-Oncogênicas c-fos/análise , Ratos , Ratos Sprague-Dawley , Medula Espinal/citologiaRESUMO
Using in vitro quantitative autoradiography and [3H]flunitrazepam we examined the rostrocaudal distribution of benzodiazepine binding sites in the human neonate/infant hypothalamus. The autoradiographic analysis shows the presence of a heterogeneous distribution throughout the rostrocaudal extent of this brain structure. High [3H]flunitrazepam binding corresponds primarily to the diagonal band of Broca and the preoptic region. The labelling in the preoptic region showed a rostrocaudal increase, contrasting in that with the other hypothalamic structures. Intermediate densities were present in the septohypothalamic, suprachiasmatic, periventricular and paraventricular nuclei as well as in the mammillary complex. Low binding was observed in the other hypothalamic structures. The benzodiazepine binding sites analyzed belong mostly to type II receptors. In an attempt to unravel possible differences related to age, we compared the autoradiographic distribution in three postnatal age ranges. The topographical distribution of these binding sites was almost identical in each period analyzed. We found, however, that benzodiazepine binding is generally low in the neonatal period and a tendency in increasing densities is observed during development. Taken together, these results provide evidence for a large distribution of benzodiazepine binding sites in neonate/infant hypothalamus, suggesting their implication in the development of this brain structure and the maintenance of its various functions.
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Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Neurônios/metabolismo , Receptores de GABA-A/metabolismo , Fatores Etários , Núcleo Hipotalâmico Anterior/citologia , Núcleo Hipotalâmico Anterior/crescimento & desenvolvimento , Núcleo Hipotalâmico Anterior/metabolismo , Ansiolíticos/farmacologia , Sítios de Ligação/efeitos dos fármacos , Sítios de Ligação/fisiologia , Feminino , Flunitrazepam/farmacocinética , Humanos , Hipotálamo/citologia , Hipotálamo Médio/citologia , Hipotálamo Médio/crescimento & desenvolvimento , Hipotálamo Médio/metabolismo , Hipotálamo Posterior/citologia , Hipotálamo Posterior/crescimento & desenvolvimento , Hipotálamo Posterior/metabolismo , Lactente , Recém-Nascido , Masculino , Neurônios/citologia , Ensaio Radioligante , Trítio/farmacocinética , Ácido gama-Aminobutírico/metabolismoRESUMO
A double-label immunofluorescence technique was used to demonstrate the co-localization of cholecystokinin-8 (CCK-8) and gonadotrophin-releasing hormone (GnRH) in individual neurons and processes of the chameleon brain. Co-localization was limited to a small population of cells in the dorsomedial tegmentum; in other regions of the brain, neurons were observed to be either CCK-8-immunopositive or GnRH-immunopositive but never both. However, double-labeled fibers and terminals were found to be distributed at a low density throughout the thalamus, the medial hypothalamus, the tegmentum and the spinal cord. These data provide the first indication for the co-localization of CCK-8 and GnRH, whose functional significance remains to be established. ON
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Hormônio Liberador de Gonadotropina/análise , Neurônios/química , Sincalida/análise , Tegmento Mesencefálico/química , Animais , Imunofluorescência , Hormônio Liberador de Gonadotropina/imunologia , Hipotálamo/química , Hipotálamo/citologia , Imuno-Histoquímica , Lagartos , Neurônios/metabolismo , Sincalida/imunologia , Tegmento Mesencefálico/citologia , Tálamo/química , Tálamo/citologiaRESUMO
After small pressure injections of Fluorogold (FG), and Dextran-tetramethylrodamine (DR) into the dorsal motor nucleus of the vagus/nucleus of the solitary tract (DMV/NTS) and the rostral ventrolateral medulla (RVLM), respectively, retrograde FG-labelled cells were found mainly in the sensorimotor cortex; retrograde DR-labelled cells were located in the same cortical areas and in the prefrontal cortex. Double-labelled cells were also found in the sensorimotor cortical areas. These results provide evidence of direct projections from the sensorimotor cortex to the DMV/NTS and RVLM and suggest that somatic cortical areas directly control cardiovascular output during sensory and somatic processes.
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Bulbo/fisiologia , Córtex Motor/fisiologia , Córtex Somatossensorial/fisiologia , Estilbamidinas , Animais , Dextranos , Corantes Fluorescentes , Histocitoquímica , Masculino , Bulbo/citologia , Córtex Motor/citologia , Vias Neurais/citologia , Vias Neurais/fisiologia , Ratos , Ratos Wistar , Rodaminas , Núcleo Solitário/citologia , Núcleo Solitário/fisiologia , Córtex Somatossensorial/citologia , Nervo Vago/fisiologiaRESUMO
The distribution of substance P-like immunoreactivity in the chameleon brain and spinal cord was studied with immunohistochemical methods using polyclonal antibodies against substance P. In the telencephalon, immunoreactive cell bodies and fibers were located primarily in the striatum and in the globus pallidus. In addition, few substance P-like fibers were observed in the cortical areas, in the septum, and in the amygdala. In the diencephalon, a high density of immunostained neurons and fibers were seen in the periventricular and ventrolateral hypothalamus. Another group of cell bodies was located in the optic tectum and particularly in the stratum griseum central. A large number of immunoreactive fibers were also detected in the thalamic nuclei and in the median eminence. In the mesencephalon, few immunoreactive neurons were observed in the ventral tegmental area, in the substantia nigra, and in the nucleus reticularis isthmi. These latter nuclei, the periventricular area, the posterior commissure, the nucleus lentiformis mesencephali, the oculomotor nucleus, and the raphe nuclei contained a dense plexus of substance P immunoreactive fibers. No immunoreactive cell bodies were observed in raphe nuclei. In the spinal cord, no substance P-like immunoreactive neurons were observed, but a large number of substance P immunostained fibers were seen in the dorsal and lateral part of the dorsal horn and surrounding the dorsal parts of the central canal. The results of the present study are discussed with respect to those obtained in other species of reptiles, the main differences concerning the lateral septum, the habenula, the area of the paraventricular organ, and the raphe nuclei.
Assuntos
Química Encefálica/fisiologia , Lagartos/fisiologia , Substância P/metabolismo , Animais , Encéfalo/anatomia & histologia , Feminino , Imuno-Histoquímica , Masculino , Medula Espinal/anatomia & histologia , Medula Espinal/imunologia , Medula Espinal/metabolismo , Substância P/imunologiaRESUMO
An immunocytochemical method, using glutaraldehyde fixation and an antiserum developed against a GABA-glutaraldehyde protein conjugate, permitted direct visualization of GABAergic structures in the brain of a reptile (chameleon). GABA immunoreactive cell bodies and nerve terminals were observed to be evenly distributed throughout the forebrain and midbrain. In the forebrain, GABA-positive perikarya were shown in all cortical areas, the septal area, the striatum, the dorsal ventricular ridge, and in the nucleus accumbens. In the midbrain, the optic tectum contained a dense and laminar distribution of GABA neurons. These neurons were also observed in the lateral geniculate nucleus, nucleus profundus mesencephali, nucleus opticus tegmenti and substantia nigra. Immunoreactive nerve fibers and terminals were observed in the same structures and, additionally, in the tractus septo-hypothalamicus, habenula complex, median eminence, intermediate lobe of the pituitary, basal part of the subcommissural organ, torus semicircularis and nucleus reticularis isthmi. These results provide a framework for a further electron microscopic analysis of the GABAergic innervation of some encephalic areas involved in physiological regulations particular to this species especially the visual system.
Assuntos
Encéfalo/citologia , Mesencéfalo/citologia , Neurônios/citologia , Ácido gama-Aminobutírico/análise , Animais , Feminino , Imuno-Histoquímica , Lagartos , Masculino , Terminações Nervosas/ultraestrutura , Fibras Nervosas/ultraestrutura , Especificidade de ÓrgãosRESUMO
Chameleons display a number of well-described physiological peculiarities of their visual system, but there is no information on the topography of the retinal ganglion cell layer. In the present study, ganglion cell density of the chameleon retina was constructed from whole mounts of the retina stained with cresyl violet. For the identification of ganglion cells, these latter cells were labelled retrogradely with horseradish peroxidase applied to the optic nerve. Using this criterion, the proportion of ganglion cells was estimated to represent 80% of retinal cells, while glial cells and amacrine cells represented 14 and 6%, respectively, of the total cell population of the retina. As for the main features of the retinal map, first, ganglion cells were distributed inhomogeneously within the ganglion cell layer, and revealed the existence of a putative area centralis. Second, a horizontal visual streak, which showed two peak density areas, was identified. These features point out the degree of specialisation of the chameleon retina and the complexity of its visual system.
Assuntos
Lagartos/anatomia & histologia , Retina/anatomia & histologia , Células Ganglionares da Retina/citologia , Animais , Peroxidase do Rábano Silvestre , Retina/citologiaRESUMO
The focus of this paper is to describe a method for the simultaneous stimulation of the vagina and uterine cervix and recording of vaginal contractions in the female rat. The influence of the estrous cycle on vaginocervical sensitivity was also investigated. The use of a latex balloon, inflated with water via a syringe and connected to a pressure transducer allowed us to record vaginal contractions, the intensity of which is an index of vaginal sensitivity, and to stimulate those spinal neurons involved in the reflex arc; at the end of the stimulation the deflated balloon was used as a probe to perform a vaginal smear in order to determine the stage of the estrous cycle at the moment of the experiment. Activated neurons were identified by Fos-immunocytochemistry. Light microscope counting of Fos-immunoreactive neurons at different stages of the estrous cycle permitted us to quantify the response to vaginocervical stimulation and to demonstrate that vaginocervical sensitivity changes significantly throughout the estrous cycle. This finding confirms the important role of sex hormones in the modulation and control of the sensitivity of the vagina and cervix, a part of the female genital tract which is implicated in a variety of neuroendocrine, behavioral and neural changes.
Assuntos
Cateterismo/instrumentação , Ciclo Estral/fisiologia , Genitália Feminina/inervação , Estimulação Física/métodos , Células do Corno Posterior/metabolismo , Transdutores de Pressão , Fibras Aferentes Viscerais/fisiologia , Animais , Cateterismo/métodos , Colo do Útero/inervação , Colo do Útero/fisiologia , Feminino , Genitália Feminina/fisiologia , Hormônios Esteroides Gonadais/metabolismo , Imuno-Histoquímica , Vértebras Lombares , Estimulação Física/instrumentação , Células do Corno Posterior/citologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Sprague-Dawley , Vagina/inervação , Vagina/fisiologiaRESUMO
The topography and localization of motoneurons innervating the six extraocular muscles in the chameleon (Chamaeleo chameleon) was studied following HRP injection in each of these individual muscles. Four muscles were innervated ipsilaterally: medial rectus, inferior rectus, inferior oblique and lateral rectus. The medial rectus muscle was innervated by the dorsomedial part of the oculomotor nucleus. The innervation to the inferior rectus muscle arose from the lateral part of the intermediate oculomotor subnucleus, which extended to the lateral part of the dorsal subdivision. The lateral rectus muscle was innervated by the abducens nucleus, which was composed by two subgroups of labeled cells, respectively observed in the principal and accessory abducens subnuclei, whereas efferents to the inferior oblique muscle originated from both the ventral and intermediate oculomotor subnuclei. The contralateral pattern consisted of motoneurons innervating the superior rectus and the superior oblique that were located respectively in the caudal portion of the ventral oculomotor nucleus and in the trochlear nucleus. These results confirmed data reported in most vertebrate species, and were discussed from a comparative and functional point of view.