Perinatal outcomes after open fetal surgery for myelomeningocele repair: a retrospective cohort study.

OBJECTIVE: Describe outcomes of open fetal surgery for myelomeningocele (MMC) repair in two Brazilian hospitals and the impact of surgical experience on outcome. DESIGN: Retrospective cohort study. SETTING: Sao Paulo, Brazil. POPULATION: 237 pregnant women carrying a fetus with an open spinal defect. METHODS: Surgical details, and maternal and fetal outcomes collected from all patients. MAIN OUTCOME MEASURES: Analysis of surgical and perinatal outcome parameters. RESULTS: Total surgical time was 119 ± 7.6 minutes. Preterm labour occurred in 24.2%, premature rupture of membranes in 26.7%, placental abruption in 0.8%, need for a blood transfusion at delivery in 2.1%, and dehiscence at the repair site in 2.5%. Reversal of hindbrain herniation at birth occurred in 71.4%. There were no maternal deaths or severe maternal morbidities. The failure rate with the patient anaesthetised was 0.42% and perinatal mortality was 2.1% (three intrauterine demises and two neonatal deaths). Comparing results from our study in the first 3 years with the last 3 years demonstrated improvement in the total surgical time (121.2 ± 6.4 versus 118.5 ± 8.2 minutes, P = 0.005) and an increase in reversal of hindbrain herniation at birth (64.0 versus 77.1%, P = 0.042). CONCLUSION: Our open fetal surgical approach for MMC was effective and results were comparable to past studies. Improvements in surgical performance and perinatal outcome increased as the surgical team became more familiar with the procedure. FUNDING: The study was funded solely by institutional funds. TWEETABLE ABSTRACT: Brazilian experience of in utero open surgery for myelomeningocele repair.

Detection of pathogenic Escherichia coli and microbiological quality of chilled shrimp sold in street markets.

UNLABELLED: Foodborne illnesses caused by Escherichia coli are one of the most important gastrointestinal diseases and therefore represent a public health risk. The presence of E. coli in water or in products such as shrimp indicates faecal contamination. However, indicator micro-organisms can be used to evaluate the microbiological quality of food sold in markets. This study focused on detecting isolates of E. coli containing the genes stx1A, stx2A, eae, LTI, STa, STb, aggR and pCVD432 in chilled shrimp sold in street markets in the municipality of São Paulo, Brazil, and to assess the microbiological quality of this product. Enteropathogenic and enterotoxigenic E. coli pathotypes were detected on the surface of two chilled shrimp samples. Salmonella spp. was not isolated. In addition, contamination of surface and muscle of the shrimp samples was found to be correlated. The detection of EPEC and ETEC pathotypes in chilled shrimp sold in street markets in Brazil provides useful epidemiological information for public health authorities to improve food safety and public health. SIGNIFICANCE AND IMPACT OF THE STUDY: Shrimps are crustaceans commonly produced and consumed in Brazil. Specimens of Farfantepenaeus brasiliensis and Litopenaeus schmitti sold in street markets were examined by PCR to detect the presence of Escherichia coli pathotypes (enteropathogenic, enterotoxigenic, enterohemorrhagic and enteroinvasive). EPEC and ETEC strains were detected in whole shrimp. These findings provide useful information for public health authorities to improve the food safety and health of the Brazilian population.

Shiga toxigenic and enteropathogenic Escherichia coli in water and fish from pay-to-fish ponds.

UNLABELLED: Escherichia coli is part of the normal microflora of the intestines of mammals. However, among the enteric pathogens, it is one of the leading causes of intestinal diseases, especially Shiga toxigenic E. coli, which can cause diarrhoea, haemorrhagic colitis and complications like haemolytic uraemic syndrome and thrombotic thrombocytopaenic purpura. Escherichia coli is considered a serious public health problem. Water and fish samples were subjected to biochemical tests to confirm the presence of E. coli and by PCR to verify the presence of pathogenic strains (O157, enteropathogenic and shiga toxigenic) in water and fish (skin, gastrointestinal tract and muscles) from pay-to-fish ponds located in the Córrego Rico watershed in the northeastern region of the state of São Paulo, Brazil. Of the 115 E. coli isolates from fish or water, five (4·34%) contained eae and stx2 genes, one had only the eae gene and two had the stx1 gene. An isolate containing the stx2 gene was also found in the water sample. In addition, eight isolates (6·95%) from the fish gastrointestinal tract contained rfbEO157:H7 (O157 gene), and three (2·61%) contained stx2 and eae genes, demonstrating the potential risk to the environment and public health. The results provide useful basic information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population. SIGNIFICANCE AND IMPACT OF THE STUDY: Pay-to-fish ponds are a common commercial activity in Brazil. Samples of water and Oreochromis niloticus were examined by PCR to detect the presence of pathogenic strains of Escherichia coli (O157, enteropathogenic and shiga toxigenic). Several pathogenic strains were detected in this study, providing useful epidemiological information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population.

Bee venom enhances the differentiation of human regulatory T cells.

Venom-specific immunotherapy (VIT) is well recognized by its efficacy, and compelling evidence implicates regulatory T cells (Tregs) in the underlying tolerogenic mechanisms. Additionally, hymenoptera venom has for a long time been claimed to modulate immunity. Here, we investigated the putative role of bee venom (Bv) in human FOXP3-expressing Treg homeostasis and differentiation, irrespective of the donors' allergic status. We found that Bv significantly enhanced the differentiation of FOXP3-expressing cells both from conventional naïve CD4 T cells and mature CD4 thymocytes, a property that may contribute to the VIT's capacity to expand circulating Tregs in allergic individuals. We expect that our data enlightening the Treg-mediated immunomodulatory properties of Bv regardless of TCR specificity, to have application in other allergies, as well as in other clinical settings, such as autoimmunity and transplantation.

Cortisol profile and clinical evaluation of canine neonates exposed antenatally to maternal corticosteroid treatment.

The effects of glucocorticoids on both foetal canine lung and endogenous serum cortisol concentration have not been clearly delineated. Therefore, we aimed to investigate whether maternal corticosteroid treatment can alter maternal and neonatal cortisol profile and improve neonatal vitality. We allocated six bitches of different breeds and their neonates into two groups: control group (CONT)--maternal administration of saline solution at 55 days post-ovulation (n = 3); and betamethasone group (BETA)--administration of a single dose of 0.5 mg/kg betamethasone (Celestone Soluspan(®) ) at 55 days post-ovulation (n = 3). Caesarean sections were scheduled for day 63 after ovulation. However, BETA group dams showed precocious signs of labour, and c-sections were performed at 58 days post-ovulation. Maternal and neonatal evaluations were performed periodically between betamethasone administration and birth, respectively. Neonates from both groups presented unsatisfactory (<5) Apgar score at birth. However, in spite of an earlier improvement on vitality found on CONT group and the premature delivery on BETA group, both groups showed acceptable Apgar score 120 min after birth. Neonatal cortisol concentrations were higher on CONT group compared to BETA group at birth. In addition, a gradual decrease on maternal cortisol concentrations was observed in the BETA group from treatment until parturition. These findings suggest that despite the down-regulation on the hypothalamic-pituitary-adrenal axis and the induction of premature delivery, betamethasone treatment was able to provide similar vitality when compared to the untreated neonates born at term.

Influence of chlorine added to drinking water during the preslaughter feed withdrawal on microbiology and morphology of the broiler gastrointestinal tract.

The objective of this experiment was to test the effects of the addition of chlorine to broiler drinking water during a 12-h preslaughter feed withdrawal period on reduction of the quantities of microorganisms, such as Escherichia coli and enterococci, in broiler crops and ceca. Reduction of these microorganisms would likely also reduce contamination of broiler meat by pathogenic bacteria during processing. It was also investigated if the chlorine caused some intestinal damage that could disseminate the microorganisms to the carcass. A total of 40 Cobb male broilers were used. Samples of crop and cecal content were collected for microbiological analysis, and duodenum and jejunum were used for morphological analysis from 10 birds in each treatment. The most probable number (MPN) of E. coli and enterococci in the collected samples of crop and ceca and the measure of the free residual chlorine in water were determined. The scanning electron microscopy from duodenum and jejunum was used to illustrate the mucosa integrity. The chlorine added to water was efficient in reducing the quantities of microorganisms in broiler crops and improved the integrity of the mucosa. Therefore, preslaughter feed withdrawal should be coupled with crop disinfection, because preslaughter feed withdrawal increases the MPN of enterococci and E. coli in broiler crops. So, it presents a higher risk for carcass contamination during slaughterhouse processing and, consequently, a higher risk for public health.

Polypharmacy among adult and older adult users of primary care services delivered through the Unified Health System in Minas Gerais, Brazil.

OBJECTIVES: To estimate the prevalence of polypharmacy (≥5 drugs) among adults and to analyze related factors. METHODS: Cross-sectional study with 1,159 interviewees distributed across 104 cities and 253 primary healthcare services delivered through the Brazilian Unified Health System. Polypharmacy-related factors were identified using logistic regression model. RESULTS: 949 (81.8%) interviewees were using at least one medication and were included in this analysis. The prevalence of polypharmacy among them was 13.7% (95%CI:11.7-16.0%) in the general population and 33.3%(95%CI:26.1-41.4%) in older adults(≥65 years). Polypharmacy was positively associated with age (45 to 64 years, OR=2.02; 95%CI:1.03-3.94; ≥65 years, OR=4.17; 95%CI:1.92-9.17) and the following chronic diseases: stroke (OR=4.20; 95%CI:1.53-11.55); diabetes mellitus (OR=4.03; 95%CI:2.43-6.68); heart disease (OR=3.18; 95%CI:1.92-5.29); depression (OR=2.85; 95%CI:1.80-4.53); hypertension (OR=2.13; 95%CI:1.17-3.86); and dyslipidemia (OR=1.73; 95%CI:1.07-2.80). CONCLUSION: This study revealed that polypharmacy is a real concern in primary health care and affects older and middle-aged adults alike. Groups of patients that are more likely to experience polypharmacy were identified. Our findings emphasize the relevance of an appropriate approach to polypharmacy driven by aging and multimorbidity.

Prediction of acidemia at birth by Doppler assessment of fetal cerebral transverse sinus in pregnancies with placental insufficiency.

OBJECTIVES: To evaluate the prediction of acidemia at birth using cerebral transverse sinus (CTS) Doppler velocimetry and to determine the best parameter and cut-off values for its prediction in pregnancies complicated with placental insufficiency. METHODS: This was a prospective cross-sectional study involving 69 pregnant women (26-40 weeks' gestation) with placental insufficiency managed in two Brazilian hospitals. Doppler assessment of the CTS was carried out in the last 24 h before delivery, and the peak ventricular systolic (S-wave) and diastolic (D-wave) velocities as well as the atrial systolic velocity (A-wave) were recorded and the pulsatility index for veins (PIV) was calculated. At birth, arterial and venous umbilical cord blood samples were collected to determine acid-base and pH status. A receiver-operating characteristics (ROC) curve was constructed for each Doppler parameter with birth acidemia as the dependent variable. Sensitivity, specificity, positive and negative predictive values, accuracy and false-positive and false-negative rates were calculated for the parameters considered to be good predictors of acidemia. RESULTS: The S, D and A peak velocities and the S/A ratio were not good predictors of acidemia at birth. The PIV and the (S - A)/S ratio were good predictors of acidemia (area under the ROC curve = 0.698 (P = 0.009) and 0.654 (P = 0.009), respectively). The cut-off values were PIV = 0.855 and (S - A)/S = 0.703). CONCLUSIONS: The PIV and the (S - A)/S ratio of the CTS were good predictors of acidemia at birth in this high-risk population with placental insufficiency.

Red wine antioxidants protect hippocampal neurons against ethanol-induced damage: a biochemical, morphological and behavioral study.

Chronic ethanol consumption increases oxidative stress, which accounts for the striking neurological changes seen in this condition. Notwithstanding, there is well-documented evidence that polyphenols, present in grape skin and seeds, exhibit a strong antioxidant activity. As red wine is rich in polyphenols, the aim of the present work was to evaluate their putative protective effects on the hippocampal formation by applying biochemical, morphological and behavioral approaches. Six-month old male Wistar rats were fed with red wine (ethanol content adjusted to 20%) and the results were compared with those from ethanol-treated (20%) rats and pair-fed controls. Biochemical markers of oxidative stress (lipid peroxidation, glutathione levels and antioxidant enzyme activities) were assessed on hippocampal homogenates. Lipofuscin pigment, an end product of lipid peroxidation, was quantified in hippocampal cornu ammonis 1 and 3 (CA1 and CA3) pyramidal neurons using stereological methods. All animals were behaviorally tested on the Morris water maze in order to assess their spatial learning and memory skills. In red wine-treated rats, lipid peroxidation was the lowest while presenting the highest levels of reduced glutathione and an induction of antioxidant enzyme activities. Morphological findings revealed that, contrary to ethanol, red wine did not increase lipofuscin deposition in CA1 and CA3 pyramidal neurons. Besides, red wine-treated animals learned the water maze task at a higher rate than ethanol group and had better performance scores by the end of the training period and on a probe trial. Actually, no significant differences were found between pair-fed controls and red wine-treated rats in morphological and behavioral data. Thus, our findings demonstrate that chronic consumption of red wine, unlike the ethanol solution alone, does not lead to a decline in hippocampal-dependent spatial memory. This may be due to the ability of red wine polyphenols to improve the antioxidant status in the brain and to prevent free radical-induced neuronal damage.

Influence of prenatal maternal corticosteroid therapy on clinical and metabolic features and pulmonary function of preterm newborn puppies.

Although the effects of antenatal corticosteroid therapy in clinical improvement and pulmonary maturation in preterm have been described, little is known on premature newborn puppies. This study aimed to evaluate the effect of maternal administration of a single dose of prenatal betamethasone on lung function of preterm newborn puppies in the first hours of life, especially from the clinical point of view and acid-base balance. A prospective study was conducted involving 21 puppies allocated into three experimental groups: Term Group (63 days post-ovulation), Preterm-Treated Group (57 days post-ovulation and maternal administration of a single dose of 0.5 mg/kg of betamethasone) and Preterm-Control Group (57 days post-ovulation). Puppies were analyzed clinically through the Apgar score, heart rate, respiratory rate and neurological tests (muscular tone and irritability reflex) and for oximetry and blood acid-base balance in distinct experimental moments. Premature puppies had marked degree of prematurity, reversed by maternal administration of betamethasone. Prenatal corticosteroid therapy promoted better pulmonary and metabolic condition, with more efficient compensatory response to acid-base imbalance and better pulmonary gas exchange capacity. Therefore, prenatal treatment with betamethasone can be adopted as clinical lung maturation protocol for pregnancies at risk in order to prevent low vitality and increase neonatal survival.

Reduced density of neuropeptide Y neurons in the somatosensory cortex of old male and female rats: relation to cholinergic depletion and recovery after nerve growth factor treatment.

Synthesis of neuropeptide Y in the neocortex and activity of the basalocortical cholinergic system are both reduced in the aging brain. We hypothesized that, by stimulating the activity of the basal forebrain cholinergic neurons, nerve growth factor might also be capable of restoring the synthesis of neuropeptide Y in cortical neurons. Old male and female rats were intraventricularly infused with nerve growth factor for 14 days and their brains were analyzed in order to quantify the densities of neuropeptide Y-immunoreactive neurons and of fiber varicosities stained for vesicular acetylcholine transporter protein in layers II/III, V and VI of the primary somatosensory barrel-field cortex. The areal densities of neuropeptide Y neurons and of vesicular acetylcholine transporter protein varicosities in all cortical laminae were found to be dramatically decreased in old rats when compared with young rats. However, infusions of nerve growth factor, known to exert a powerful trophic effect upon cortically projecting cholinergic neurons, have led to considerable recovery of vesicular acetylcholine transporter protein-positive terminal fields, which was paralleled by complete restoration of function in neuropeptide Y-producing neurons. With respect to the gender differences, although the density of cortical neuropeptide Y neurons was found to be significantly higher in young females than in young males and the opposite was true for vesicular acetylcholine transporter protein-positive varicosities, the general pattern of age- and treatment-related changes in these neurochemical markers was similar in both sexes. Overall, the age- and treatment-related variations in the density of cortical neuropeptide Y cells were found to correlate with those observed in the density of vesicular acetylcholine transporter protein varicosities. These results lend support to the idea that there is a causal relationship between age-related changes in cortical cholinergic and neuropeptide Y-ergic neurotransmitter systems.

Anterior vocal commissure invasion in laryngeal carcinoma diagnosis.

BACKGROUND: Laryngeal carcinoma involving anterior vocal commissure (AVC) represents a great challenge for staging and treatment. OBJECTIVES: To compare laryngoscopy and computed tomography (CT) scan efficiency in staging tumors extending to the AVC. We also analyzed the helicoidal axial CT scan accuracy in recognizing this larynx subregion invasion. MATERIAL AND METHODS: Fifty-two glottic and supraglottic laryngeal squamous cell carcinoma patients with tumoral extension to the AVC were prospectively studied from August 2001 to August 2003 at the National Cancer Institute (Rio de Janeiro, Brazil). All patients underwent videolaryngoscopic examination and direct laryngoscopy for lesion extension analysis and biopsy. After AVC helicoidal axial CT scan with sagittal and coronal 1.0 mm thick reconstruction, patients were submitted to surgical treatment. The same pathologist analyzed all surgical specimens. RESULTS: When compared with pathologic stage, clinical endoscopic classification was correct in 40.38% of cases (40% for T1, 29.41% for T2, 46.43% for T3, and 50% in T4). Helicoidal axial CT scan accuracy for AVC tumors was 75% (P = .0001), being more important for T2 (62.50%), T3 (73.91%), and T4 (88.24%) lesions. Identification of radiologic signs described as gross radiologic anterior commissure involvement (GRACI) increased radiologic image staging accuracy to over 96%. CONCLUSIONS: Endoscopic evaluation understaged tumors in all clinical stages but really T1. Helicoidal axial CT scan reformatted to 1.0 mm thick played an important role in correctly staging more advanced AVC laryngeal tumors. Radiologic signs, here identified as GRACI, may be very helpful for tomographic staging and patient treatment.

Lesions of the laterodorsal tegmental nucleus alter the cholinergic innervation and neuropeptide Y expression in the medial prefrontal cortex and nucleus accumbens.

The effects of the ibotenic acid infused into the area of the laterodorsal tegmental nucleus (LDT) of rats on the expression of cortical and accumbal neuropeptides were assessed. The effects of this manipulation were determined in the nucleus accumbens (NAc) and medial prefrontal cortex (mPFC) by estimating the numerical density of varicosities immunoreactive for vesicular acetylcholine transporter and the total number of NAc neurons immunoreactive for choline acetyltransferase (ChAT) and neuropeptide Y (NPY) as well as the total number of mPFC neurons immunoreactive for NPY and vasoactive intestinal polypeptide (VIP). In LDT-lesioned rats, the density of the cholinergic varicosities was reduced in the ventral divisions of the mPFC and in all divisions of the NAc. In addition, in these rats, the total number of NPY-immunoreactive neurons was reduced in all subregions of the mPFC and in the NAc. Conversely, the total number of VIP-immunoreactive neurons in the mPFC and of ChAT-immunoreactive neurons in the NAc did not differ between LDT- and sham-lesioned rats. These data provide the first direct evidence for a relationship between selective damage of LDT cholinergic neurons and decreased expression of NPY in the mPFC and NAc. They also reveal that different types of cortical and accumbal interneurons respond differently to the cholinergic denervation induced by LDT lesions.

Chronic alcohol consumption induces lipofuscin deposition in the rat hippocampus.

Previous investigations have suggested that chronic alcohol consumption accelerates a number of age-related changes in the cerebellar cortex and hippocampal formation. In the cerebellum, alcohol-feeding has been shown to accelerate the intracellular deposition of lipofuscin. In order to determine whether alcohol administration has a similar effect on hippocampal lipofuscin deposition, we studied the pattern of lipofuscin deposition in alcohol-fed rats for periods of 1, 3, 6, 12 and 18 months and compared the results with those obtained in the respective pair-fed controls. A precocious and progressive deposition of lipofuscin pigment was found in both CA1 and CA3 neurons in Ammon's horn hippocampal fields after 3 and 6 months of alcohol feeding, respectively. These results parallel those observed during normal aging and reinforce the hypothesis of a close link between chronic alcohol consumption and a premature nerve cell aging.

Reorganization of mossy fiber synapses in male and female hypothyroid rats: a stereological study.

Thyroid hormone deficiency has been reported to interfere with synaptogenesis, particularly in those regions of the brain where the neurons display a late and protracted histogenesis, although the extent of the synaptic alterations remains unknown. To provide detailed quantitative data on the effects of hypothyroidism upon synapses, a link of the hippocampal circuitry was selected: the contact between mossy fibers and dendritic excrescences of CA3 pyramidal cells (MF-CA3 synapses). Groups of six male and six female rats aged 30 and 180 days were analyzed separately after being treated as follows: (1) hypothyroid from day 0 until day 30 (30 day old hypothyroid group); (2) hypothyroid from day 0 until day 180 (180 day old hypothyroid group); (3) hypothyroid until day 30 and thenceforth maintained euthyroid (recovery group); and (4) and (5) 30 and 180 day old control groups, respectively. Timm staining, Golgi impregnation, and electron microscopy were employed to estimate the volume of the mossy fiber system, the number and size of mossy fiber boutons, and the number and related features of MF-CA3 synapses. The volume of the mossy fiber system and the number of synaptic boutons were reduced in all experimental groups. The total number of synapses was decreased in 30 day old hypothyroid rats, but did not differ among 180 day old animals. Postsynaptic densities were shorter in hypothyroid and recovery groups than in controls, although the reduction was not as marked in recovery rats as it was in hypothyroid animals. Structural alterations were noted in the pre- and postsynaptic compartments of MF-CA3 synapses of both 180 day old hypothyroid and recovery rats. These changes can be regarded as mechanisms of reorganization as they underlie the compensation for the hypothyroid-induced numerical reduction of synapses observed in 30 day old animals and enable a complete catch-up of their total number. However, synaptic reorganization was not fully achieved, as revealed by the reduction in the size of the synaptic sites in hypothyroid and recovery animals. Finally, we demonstrate that hypothyroidism did not interfere with the sex-related differences of MF-CA3 synapses described in normal rats.

An ultrastructural morphometric study of mossy fiber endings in pigeon, rat and man.

Mossy fiber endings and their synapses have an increasing morphological complexity along the phylogenetic scale which is specially evident when the evolution from reptiles to birds is considered. Among birds and mammals, only scanty and merely qualitative differences have been noticed in the mossy fiber organization. In the present study, the mossy fiber endings organization within the neuropil of four pigeons, four rats and six men was studied at optic and ultrastructural levels using morphometric methods. No significant differences were found in the fraction of volume of the neuropil and in the number of granule cells per unit volume of granular layer in all the three species. In man, a significant decrease in the volume of mossy fiber endings per unit volume of neuropil, and significant increases in their surface-to-volume ratio, in the fraction of their neurolemma occupied by synaptic contacts and in the average length of their synaptic contact zones, were found. The total synaptic area of mossy fiber endings per unit volume of neuropil was larger in man than in other animals in spite of their more reduced volume. These results show that significant differences in mossy fiber organization can be demonstrated when man is compared with a small mammal (the rat) and a bird (the pigeon). It is suggested that differences in the stereological organization of man's terminals might be related to his capacity for making finely graded movements. Moreover, it is suggested that the phylogenetic evolution of the mossy fiber organization has progressed among the higher vertebrates as it did in the rest of the animal scale.

Influence of sex and estrus cycle on the sexual dimorphisms of the hypothalamic ventromedial nucleus: stereological evaluation and Golgi study.

Neurons in the ventromedial nucleus of the hypothalamus (VMN) display structural and biochemical sex differences in response to estrogen. Despite this fact, reports on sex differences in the morphology of the VMN are restricted to its volume and synaptic patterning. The aim of this study was to characterize the neuroanatomical sexual dimorphisms in the VMN and to investigate whether endogenous changes in ovarian steroid secretion influence such dimorphisms. The VMN of adult male rats and intact, aged-matched female rats killed on proestrus and diestrus day 1 was examined by using stereological methods applied to conventionally stained sections and Golgi-impregnated material. The VMN contained 55,000 neurons in rats of both sexes, but its volume was, on average, 1.25 times larger in males than in females. The volume was greater in proestrus than in diestrus rats due to parallel changes in the neuronal somatic size. Unlike the dorsomedial division, neurons in the ventrolateral division had longer dendritic trees in proestrus than in diestrus females and males. The spine density was consistently higher in females than in males in both VMN divisions. In addition, in the ventrolateral part the magnitude of the sex differences varied across the estrus cycle, and reached the greatest value when females were in proestrus. The volume of the neuropil was significantly larger in males than in females, and was not affected by the estrus phase. Our results reveal that the magnitude of the neuroanatomical sex differences in the VMN vary across the estrus cycle due to the trophic influence of estrogen upon its neurons. They also show that the fundamental sex difference in the structure of the VMN is accounted for by the neuropil components.

Age and sex do not affect the volume, cell numbers, or cell size of the suprachiasmatic nucleus of the rat: an unbiased stereological study.

The circadian rhythms displayed by numerous biological functions are known to be sex specific and affected by aging. It has not been settled yet whether the sex- and age-related characteristics of circadian rhythms derive from changes in the anatomy of the suprachiasmatic nucleus. To shed light on these issues, we applied unbiased stereological techniques to estimate the volume of the suprachiasmatic nucleus as well as the total number of its cells and the mean volume of their somata and nuclei in progressively older groups of male and female Wistar rats (aged 1, 6, 12, 18, 24, and 30 months). The volume of the nucleus was estimated with the Cavalieri principle on serial sections. The total numbers of neurons and astrocytes were estimated by applying the optical fractionator, and the mean somatic and nuclear volumes of cells were estimated by using isotropic, uniform random sections and the nucleator method. On average, the volume of the suprachiasmatic nucleus was 0.044 mm3, and the total number of neurons and astrocytes was 17,400. Cells of the dorsomedial and ventrolateral components of the nucleus, which are morphologically different, have identical mean perikaryal and nuclear volumes, which we estimated to be 750 microns3 and 400 microns3, respectively. We further demonstrated that, at all ages analysed, the volume of the suprachiasmatic nucleus, the total cell number, and the mean somatic and nuclear volumes of its cells are affected neither by the age nor by the sex of the animal, regardless of the presence of sex- and age-related variations in circadian rhythms. However, the possibility that females may display changes in the volume of the suprachiasmatic nucleus at older ages cannot be ruled out. No effect of aging was observed in the total number of neurons or in the total number of astrocytes.

Effects of hypothyroidism upon the granular layer of the dentate gyrus in male and female adult rats: a morphometric study.

The effects of hypothyroidism upon the structure of the central nervous system of adult rats are poorly understood in spite of evidence that the mature brain is vulnerable to this condition. Existing developmental studies show that the morphological changes induced by thyroid hormone deficiency are related to alterations in neurogenesis. We studied the granular layer of the dentate gyrus under different experimental conditions of hypothyroidism, because in rodents the neurogenesis of the granule cells continues during adulthood. The following groups of rats were analysed: 1) control; 2) hypothyroid from day 0 until day 180 (hypothyroid group); 3) hypothyroid until day 30 and henceforth maintained euthyroid (recovery group); and 4) hypothyroid since day 30 (adult hypothyroid group). Groups of 6 male rats and 6 female rats were analysed separately. The volume of the dentate gyrus granular layer and the numerical density of its neurons were evaluated, so we were able to estimate the total number of granule cells. Because in the experimental groups the volume of the granular layer and the numerical density of its neurons were reduced, the total number of granule cells was decreased. In the hypothyroid and recovery groups the alterations were identical and more striking than in the adult hypothyroid groups. The total number of granule cells displayed sexual differences in all groups studied except in the hypothyroid groups. The present results support the view that thyroid hormone deficiency interferes with the process of cell acquisition by reducing neuronal proliferation and that it also leads to increased cell death. These events underlie the irreversible morphological changes observed in the brain of hypothyroid rats, either during development or at maturity. The referred structural alterations are probably related to the functional deficits observed in this condition.

Arcuate nucleus of the hypothalamus: effects of age and sex.

The arcuate nucleus of the hypothalamus (ARN) is involved in a variety of functions known to be sexually dimorphic and altered by aging. Although the effects of sex and age on the synaptic organization and neurochemistry of the ARN have been extensively analyzed, data regarding sex-related differences and age-induced effects on the total number of neurons and volume of the ARN in adult and aged male and female rats are controversial. To address this issue, we have quantitatively analyzed the ARN of male and female Wistar rats aged 6 and 24 months. The optical fractionator, the optical rotator, and the Principle of Cavalieri were used as the estimators of the total number of neurons, mean nuclear volume of ARN neurons, and volume of the ARN, respectively. In addition, a Golgi study was carried out to analyze the dendritic trees of its neurons. We found that in young adult rats, the volume of the ARN is 0.9 mm3 in males and 0.7 mm3 in females, whereas the total number of neurons is 100 x 10(3) in males and 86 x 10(3) in females. ARN neurons of males and females have identical mean nuclear volumes, which we estimated to be 300 microm3. No significant effects of age were found in these parameters, both in males and in females. In adult rats, no sex-related differences were detected in the number of dendritic segments and in the total dendritic length, but the dendritic branching density and the spine density were greater in females than in males. In aged rats there was a significant reduction in the number of dendritic segments, in the total dendritic length, and in the branching and spine densities that, although evident in both sexes, was more marked in females. Our results show that the total number of neurons and the volume of the ARN are sexually dimorphic in adult and aged rats and that neither of these parameters is altered by aging. Conversely, aging induces regressive changes in the dendritic arborizations of ARN neurons of males and females and abolishes the sexual dimorphic pattern of their organization.