Pre, intra and post-ischemic hypothermic neuroprotection in temporary focal cerebral ischemia in rats: morphometric analysis

Objective: To evaluate the neuroprotection of mild hypothermia, applied in different moments, in temporary focal cerebral ischemia in rats. Methods: Rats was divided into Control (C), Sham (S), Ischemic-control(IC), Pre-ischemic Hypothermia (IH1), Intra-ischemic Hypothermia (IH2), and Post-ischemic Hypothermia (IH3) groups. Morphometry was performed using the KS400 software (Carl Zeiss (R)) in coronal sections stained by Luxol Fast Blue. Ischemic areas and volumes were obtained. Results: Statistically, blue areas showed difference for C vs. IC, IC vs. IH1 and IC vs. IH2 (p=0.0001; p=0.01; p=0.03), and no difference between C vs. S, IC vs. IH3 and IH vs. IH2 (p=0.39; p=0.85; p=0.63). Red areas showed difference between C vs. IC, IC vs. IH1 and IC vs. IH2 (p=0.0001; p=0.009; p=0.03), and no difference between C vs. S, IC vs. IH3 and IH1 vs. IH2 (p=0.48; p=0.27; p=0.68). Average ischemic areas and ischemic volumes showed difference between IC vs. IH1 and IC vs. IH2 (p=0.0001 and p=0.0011), and no difference between IC vs. IH3 and IH1 vs. IH2 (p=0.57; p=0.79). Conclusion: Pre-ischemic and intra-ischemic hypothermia were shown to be similarly neuroprotective, but this was not true for post-ischemic hypothermia.

Morphometric analysis of the placenta in the New World mouse Necromys lasiurus (Rodentia, Cricetidae): a comparison of placental development in cricetids and murids

Background: Stereology is an established method to extrapolate three-dimensional quantities from two-dimensional images. It was applied to placentation in the mouse, but not yet for other rodents. Herein, we provide the first study on quantitative placental development in a sigmodontine rodent species with relatively similar gestational time. Placental structure was also compared to the mouse, in order to evaluate similarities and differences in developmental patterns at the end of gestation. Methods: Fetal and placental tissues of Necromys lasiurus were collected and weighed at 3 different stages of gestation (early, mid and late gestation) for placental stereology. The total and relative volumes of placenta and of its main layers were investigated. Volume fractions of labyrinth components were quantified by the One Stop method in 31 placentae collected from different individuals, using the Mercator® software. Data generated at the end of gestation from N. lasiurus placentae were compared to those of Mus musculus domesticus obtained at the same stage. Results: A significant increase in the total absolute volumes of the placenta and its main layers occurred from early to mid-gestation, followed by a reduction near term, with the labyrinth layer becoming the most prominent area. Moreover, at the end of gestation, the total volume of the mouse placenta was significantly increased compared to that of N. lasiurus although the proportions of the labyrinth layer and junctional zones were similar. Analysis of the volume fractions of the components in the labyrinth indicated a significant increase in fetal vessels and sinusoidal giant cells, a decrease in labyrinthine trophoblast whereas the proportion of maternal blood space remained stable in the course of gestation. On the other hand, in the mouse, volume fractions of fetal vessels and sinusoidal giant cells decreased whereas the volume fraction of labyrinthine trophoblast increased compared to N. lasiurus placenta. Conclusions: Placental development differed between N. lasiurus and M. musculus domesticus. In particular, the low placental efficiency in N. lasiurus seemed to induce morphological optimization of fetomaternal exchanges. In conclusion, despite similar structural aspects of placentation in these species, the quantitative dynamics showed important differences.

Morphometric analysis of fetal development of Cavia porcellus (Linnaeus, 1758) by ultrasonography: pilot study

Measurements on the growth process and placental development of the embryo and fetuses of Cavia porcellus were carried out using ultrasonography. Embryo, fetus, and placenta were monitored from Day 15 after mating day to the end of gestation. Based on linear and quadratic regressions, the following morphometric analysis showed a good indicator of the gestational age: placental diameter, biparietal diameter, renal length, and crown rump. The embryonic cardiac beat was first detected at an average of 22.5 days. The placental diameter showed constant increase from beginning of gestation then remained to term and presented a quadratic correlation with gestational age (r2 = 0.89). Mean placental diameter at the end of pregnancy was 3.5 ± 0.23 cm. By Day 30, it was possible to measure biparietal diameter, which followed a linear pattern of increase up to the end of gestation (r2 = 0.95). Mean biparietal diameter in the end of pregnancy was 1.94 ± 0.03 cm. Kidneys were firstly observed on Day 35 as hyperechoic structures without the distinction of medullar and cortical layers, thus the regression model equation between kidney length and gestational age presents a quadratic relationship (r2 = 0.7). The crown rump presented a simple linear growth, starting from 15 days of gestation, displaying a high correlation with the gestational age (r2 = 0.9). The offspring were born after an average gestation of 61.3 days. In this study, we conclude that biparietal diameter, placental diameter, and crown rump are adequate predictive parameters of gestational age in guinea pigs because they present high correlation index.

Spatial analysis of leprosy incidence and associated socioeconomic factors

OBJECTIVE: To identify clusters of the major occurrences of leprosy and their associated socioeconomic and demographic factors. METHODS: Cases of leprosy that occurred between 1998 and 2007 in Sao Jose do Rio Preto (southeastern Brazil) were geocodified and the incidence rates were calculated by census tract. A socioeconomic classification score was obtained using principal component analysis of socioeconomic variables. Thematic maps to visualize the spatial distribution of the incidence of leprosy with respect to socioeconomic levels and demographic density were constructed using geostatistics. RESULTS: While the incidence rate for the entire city was 10.4 cases per 100,000 inhabitants annually between 1998 and 2007, the incidence rates of individual census tracts were heterogeneous, with values that ranged from 0 to 26.9 cases per 100,000 inhabitants per year. Areas with a high leprosy incidence were associated with lower socioeconomic levels. There were identified clusters of leprosy cases, however there was no association between disease incidence and demographic density. There was a disparity between the places where the majority of ill people lived and the location of healthcare services. CONCLUSIONS: The spatial analysis techniques utilized identified the poorer neighborhoods of the city as the areas with the highest risk for the disease. These data show that health departments must prioritize politico-administrative policies to minimize the effects of social inequality and improve the standards of living, hygiene, and education of the population in order to reduce the incidence of leprosy.

Population divergence of Melipona scutellaris (Hymenoptera: Meliponina) in two restricted areas in Bahia, Brazil

Melipona scutellaris Latreille has great economic and ecological importance, especially because it is a pollinator of native plant species. Despite the importance of this species, there is little information about the conservation status of their populations. The objective of this study was to assess the diversity in populations of M. scutellaris coming from a Semideciduous Forest Fragment and an Atlantic Forest Fragment in the Northeast Brazil, through geometric morphometric analysis of wings in worker bees. In each area, worker bees were collected from 10 colonies, 10 workers per colony. To assess the diversity on the right wings of worker bees, 15 landmarks were plotted and the measures were used in analysis of variance and multivariate analysis, principal component analysis, discriminant analysis and clustering analysis. There were significant differences in the shape of the wing venation patterns between colonies of two sites (Wilk's lambda = 0.000006; p < 0.000001), which is probably due to the geographical distance between places of origin which impedes the gene flow between them. It indicates that inter and intrapopulation morphometric variability exists (p < 0.000001) in M. scutellaris coming from two different biomes, revealing the existence of diversity in these populations, which is necessary for the conservation of this bee species.

Spatial regimes: dynamics of intentional homicides in the city of Sao Paulo between 2000 and 2008

Objective. To identify the existence of spatial and temporal patterns in the occurrence of intentional homicides in the municipality of Sao Paulo (MSP), Brazil, and to discuss the analytical value of taking such patterns into account when designing studies that address the dynamics and factors associated with the incidence of homicides. Methods. A longitudinal ecological study was conducted, having as units of analysis 13 205 census tracts and the 96 census districts that congregate these sectors in Sao Paulo. All intentional homicides reported in the city between 2000 and 2008 were analyzed. The gross homicide rates per 100 000 population was calculated as well as the global and local Bayesian estimates for each census tract during the study period. To verify the possibility of identifying different patterns of the spatial distribution of homicides, we used BoxMap and Moran's I index. Results. The homicide trends in the city of Sao Paulo in the last decade were not homogeneous and systematic. Instead, seven patterns of spatial distribution were identified; that is, seven spatial regimes for the occurrence of intentional homicides, considering the homicide rates within each census tract as well as the rates in adjacent tracts. These spatial distribution regimes were not contained within the limits of the census tracts and districts. Conclusions. The results show the importance of analyzing the spatial distribution of social phenomena without restriction of political and administrative boundaries.

Analysis of the NMDA in Focal Cerebral Ischemia in Rats

NMDAR (N-methyl-D-aspartate receptor) is one subtype of ionotrophic glutamate receptor which is extensively distributed in the central nervous system (CNS). In the mammalian CNS, NMDAR serves prominent roles in the pathophysiologic process of cerebral ischemia. This study aimed to investigate the pattern of expression of protein and gene of the excitatory neurotransmitter NMDAR in experimental focal cerebral ischemia and the hole of neuroprotection with hypothermia and ketoprofen. 120 rats were randomly divided into 6 groups (20 animals each): control - no surgery; sham - simulation of surgery; ischemic - focal ischemia for 1 hour, without reperfusion; ischemic + intraischemic hypothermia; ischemic + previous intravenous ketoprofen, and ischemic + hypothermia and ketoprofen. Ten animals from each experimental group were used to establish the volume of infarct. Transient focal cerebral ischemia was obtained in rats by occlusion of the middle cerebral artery with an intraluminal suture. The infarct volume was measured using morphometric analysis of infarct areas defined by triphenyl tetrazolium chloride and the patterns of expression of the protein and gene NMDA were evaluated by immunohistochemistry and quantitative real-time PCR, respectively. Increases in the protein and gene NMDA receptor in the ischemics areas were observed and these increases were reduced by hypothermia and ketoprofen. The increase in the NMDA receptor protein and gene expression observed in the ischemic animals was reduced by neuroprotection (hypothermia and ketoprofen). The NMDA receptor increases in the ischemic area suggests that the NMDA mediated neuroexcitotoxicity plays an important role in cell death and that the neuroprotective effect of both, hypothermia and ketoprofen is directly involved with the NMDA.

Morphological aspects of the ovaries of turtle Kinosternon scorpioides raised in captivity

Chaves E.P., Oliveira S.C.R., Araujo L.P.F., Oliveira A.S., Miglino M.A., Abreu-Silva A.L., Melo F.A. & Sousa A.L. 2012. Morphological aspects of the ovaries of turtle Kinosternon scorpioides raised in captivity. Pesquisa Veterinaria Brasileira 32(7):667-671. Departamento das Clinicas, Curso de Medicina Veterinaria, Universidade Estadual do Maranhao, Cidade Universitaria Paulo VI, Tirirical, Sao Luis, MA 65050-150, Brazil. E-mail: alana@elo.com.br The swear turtle "jurara" (Kinosternon scorpioides) is a mud turtle of the Amazon region exposed to disordering capture in the rural areas of Maranhao, Brazil. Despite its popularity in these areas, little meaningful information regarding the reproductive morphology is currently available, fact that impedes the adoption of policies for preservation of the species. To obtain more information, we studied the ovarian morphology adult jurara females kept in captivity by morphological and morphometric analysis in the dry and rainy season. The results revealed that all females were sexually mature and were in a vitellogenic period. The ovaries are two irregular structures composed by follicles in different stages of development (primary, secondary and tertiary) scattered in a stroma of loose connective highly vascularized tissue. The ovary weight was 6.25+/-4.23g and 2.27+/-1.42g, for the right and left one respectively. The gonadosomatic indexes were 2.06% for the dry season and 1.79% for the rainy season. The average of the follicles was 29.83 units per ovary. Microscopically, the mature ovaries revealed a basal layer composed by four cellular layers: the inner and outer theca, stratum granulosum with perivitelline membrane and zona radiata with vitelline membrane. No significant differences were observed in the ovaries either in the dry or wet period.

Comparison of wing geometry data and genetic data for assessing the population structure of Aedes aegypti

Aedes aegypti is the most important vector of dengue viruses in tropical and subtropical regions. Because vaccines are still under development, dengue prevention depends primarily on vector control. Population genetics is a common approach in research involving Ae. aegypti. In the context of medical entomology, wing morphometric analysis has been proposed as a strong and low-cost complementary tool for investigating population structure. Therefore, we comparatively evaluated the genetic and phenotypic variability of population samples of Ae. aegypti from four sampling sites in the metropolitan area of Sao Paulo city, Brazil. The distances between the sites ranged from 7.1 to 50 km. This area, where knowledge on the population genetics of this mosquito is incipient, was chosen due to the thousands of dengue cases registered yearly. The analysed loci were polymorphic, and they revealed population structure (global F-ST = 0.062; p < 0.05) and low levels of gene flow (Nm = 0.47) between the four locations. Principal component and discriminant analyses of wing shape variables (18 landmarks) demonstrated that wing polymorphisms were only slightly more common between populations than within populations. Whereas microsatellites allowed for geographic differentiation, wing geometry failed to distinguish the samples. These data suggest that microevolution in this species may affect genetic and morphological characters to different degrees. In this case, wing shape was not validated as a marker for assessing population structure. According to the interpretation of a previous report, the wing shape of Ae. aegypti does not vary significantly because it is stabilised by selective pressure. (C) 2011 Elsevier B.V. All rights reserved.

Assessment of field fertility and several in vitro sperm characteristics following the use of different Angus sires in a timed-AI program with suckled Nelore cows

In order to provide information that may help researchers to understand the main cause(s) of differences in bull fertility frequently observed in field trials, this study aimed to investigate conception rates as well as several in vitro sperm characteristics of different sires of unknown fertility utilized in a Timed-AI (TAI) program. Suckled Nelore cows submitted to the same TAI protocol were allocated into eight breeding groups of approximately 120 animals each. Frozen semen doses from three Angus bulls and three different batches from each bull were utilized. Approximately 100 doses from each batch were used in TAI. Sires, batches and AI technicians were equally distributed across breeding groups. Cows were examined for pregnancy diagnosis 40 d after TAI. For in vitro sperm analyses, the same thawing procedure was repeated in the laboratory to mimic field conditions. The following in vitro sperm characteristics were assessed: computerized motility, thermal resistance, plasma and acrosomal membrane integrity, lipid peroxidation, morphology, morphometry and chromatin structure. No effect of breeding group, body condition score, AI technician and sire was observed. However, some significant differences among bulls were detected in laboratory analyses. Semen from sire presenting numerically lower (P > 0.05) pregnancy/AI also presented lower (P < 0.05) values in all sperm characteristics analyzed in thermal resistance test at 4 h (Total Motility, Progressive Motility, Average Path Velocity, Straight-Line Velocity, Curvilinear Velocity, Amplitude of Lateral Head Displacement, Beat Cross Frequency, Straightness, Linearity, and Percentage of Rapidly Moving Cells), higher (P < 0.05) Major and Total Defects in sperm morphological test, lower (P < 0.05) Length, Ellipticity and Fourier parameter (Fourier 0) in sperm morphometric analysis as well as higher (P < 0.05) chromatin heterogeneity. It was concluded that, although no bull effect was observed in the field experiment, the sire that presented numerically lower pregnancy/AI also presented lower semen quality according to the laboratory analyses performed. (C) 2012 Elsevier B.V. All rights reserved.

Associação de variáveis ambientais à ocorrência de leptospirose canina e humana na cidade de São Paulo

Foi realizada uma análise espacial da ocorrência de leptospirose humana e canina na Supervisão de Vigilância em Saúde do Butantã, situada no município de São Paulo, no ano de 2007, associada a variáveis ambientais de risco, tais como: focos de enchente e áreas de desratização. Foram encontrados aglomerados espaciais de pontos de alagamentos em 12 setores censitários e de casos de leptospirose humana em quatro setores censitários, sem correlação entre ambos. Não foram encontrados agrupamentos de casos em cães, possivelmente devido à subnotificação. As proporções casos humanos de leptospirose : população humana dentro e fora da área de desratização foram 7:199.600 e 9:257.980, respectivamente. Conclui-se que medidas de controle de roedores como a desratização foram responsáveis pela minimização dos efeitos dos fatores de risco para a transmissão de leptospirose para humanos.

Mapeamento de solos e da produtividade de plantações de Eucalyptus grandis, com uso de sistema de informação geográfica

Detailed environmental land characterization is essential for technical and financial planning, for both the scientific point of view and technological application. This work aimed at the physiographic and pedological characterization and eucalyptus productivity mapping at Itatinga Forest Sciences Experimental Station (southeastern Brazil), using geographic information systems in order to identify possible cause-effect relationships between forest productivity and soil attributes. The digital cartographic dataset was structured as follows: as primary source of data, aerial photograph and field survey were used and, as a secondary source, topographical, geological and land use occupation maps were used. For mapping wood productivity at age six (MAI6, Mean Annual Increment), inventory data of permanent plots (same species, provenance and age) were used, which were obtained from Eucalyptus grandis plantations. Using simple linear correlation and backward stepwise multiple regression analysis, the dependent variable (MAI) was related with physical and chemical characteristics of the soils. Two standards of contour curves were identified, one with close curves, narrow and surrounding the drainage network, in the steeper and lower altitude areas; the other, with spaced contour lines, in the areas of higher altitude and with plane relief. Six types of soils were characterized as being highly related to the physiographic patterns of the area: loamy sandy to sandy clayey Typic Hapludox (LVAd, 47.5%), clayey Rhodic Hapludox (LVd1, 33.4%), sandy clay Rhodic Hapludox (LVd2, 6%), clayey Rhodic Hapludox (LVdf, 9.1%), Entisols (G, 3.4%) and Fluvents soil (RY, 0.6%). There were large variations in wood productivity in the Eucalyptus grandis plantations, characterized in six classes, ranging from 26 to 52 m(3) ha(-1) yr(-1). These productivity changes were strictly related to soil mapping units. Through multiple regression analysis, we found that clay and organic matter contents were the attributes which most strongly explained the productivity differences.

Sural nerve involvement in experimental hypertension: morphology and morphometry in male and female normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR)

Background: The sural nerve has been widely investigated in experimental models of neuropathies but information about its involvement in hypertension was not yet explored. The aim of the present study was to compare the morphological and morphometric aspects of different segments of the sural nerve in male and female spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. Rats aged 20 weeks (N = 6 in each group) were investigated. After arterial pressure and heart rate recordings in anesthetized animals, right and left sural nerves were removed and prepared for epoxy resin embedding and light microscopy. Morphometric analysis was performed with the aid of computer software, and took into consideration the fascicle area and diameter, as well as myelinated fiber number, density, area and diameter. Results: Significant differences were observed for the myelinated fiber number and density, comparing different genders of WKY and SHR. Also, significant differences for the morphological (thickening of the endoneural blood vessel walls and lumen reduction) and morphometric (myelinated fibers diameter and G ratio) parameters of myelinated fibers were identified. Morphological exam of the myelinated fibers suggested the presence of a neuropathy due to hypertension in both SHR genders. Conclusions: These results indicate that hypertension altered important morphometric parameters related to nerve conduction of sural nerve in hypertensive animals. Moreover the comparison between males and females of WKY and SHR allows the conclusion that the morphological and morphometric parameters of sural nerve are not gender related. The morphometric approach confirmed the presence of neuropathy, mainly associated to the small myelinated fibers. In conclusion, the present study collected evidences that the high blood pressure in SHR is affecting the sural nerve myelinated fibers.

Análise espacial de óbitos por acidentes de trânsito, antes e após a Lei Seca, nas microrregiões do estado de São Paulo

OBJETIVO: Identificar aglomerados espaciais de microrregiões segundo taxas de óbito por acidentes de trânsito, no Estado de São Paulo, 1 ano antes e 1 ano após a Lei Seca. MÉTODOS: Estudo ecológico e exploratório, no período de 2007 e 2009, em 63 microrregiões do Estado de SP, Brasil. Utilizaram-se ferramentas de geoprocessamento com dados do DATASUS; analisando óbitos decorrentes de acidentes de trânsito em taxas por 100 mil habitantes, construindo coropletes. Nova estatística foi obtida subtraindo-se a taxa de 2009 de 2007, observando regiões de melhora ou piora. RESULTADOS: Em 2007, ocorreram 5.204 óbitos, com média de 83 óbitos/microrregião, variando entre 1 e 1.440. Já 2009 obteve 5.065 óbitos com a média de 80 óbitos/microrregião, variando entre 1 e 1.453. O coeficiente de Moran em 2007 foi I = 0,09 (p = 0,04) com correlação espacial positiva e 2009 obteve I = 0,04 (p = 0,16), sem correlação. A diferença entre as taxas obteve I = 0,23 (p = 0, 007), indicando associação espacial. Em 2007, Presidente Prudente, Rio Claro, Campinas, Bragança Paulista, Osasco e São Paulo tiveram altas taxas de óbito. Dessas regiões citadas, somente Osasco não se destacou em 2009. Ribeirão Preto, Ourinhos e Avaré obtiveram piora em 2009. A diferença entre as taxas de 2009 e 2007 mostrou que Amparo, Bragança Paulista e Campinas tiveram melhora, e Presidente Prudente e Ourinhos apresentaram piora. CONCLUSÃO: Foi possível identificar os locais com as maiores taxas de mortalidade, apontando locais onde as ações de fiscalização devem ser revisadas.