Characterization of Neural Stem/Progenitor Cells Expressing VEGF and its Receptors in the Subventricular Zone of Newborn Piglet Brain

Neural stem/progenitor cell (NSP) biology and neurogenesis in adult central nervous system (CNS) are important both towards potential future therapeutic applications for CNS repair, and for the fundamental function of the CNS. In the present study, we report the characterization of NSP population from subventricular zone (SVZ) of neonatal piglet brain using in vivo and in vitro systems. We show that the nestin and vimentin-positive neural progenitor cells are present in the SVZ of the lateral ventricles of neonatal piglet brain. In vitro, piglet NSPs proliferated as neurospheres, expressed the typical protein of neural progenitors, nestin and a range of well-established neurodevelopmental markers. Upon dissociation and subculture, piglet NSPs differentiated into neurons and glial cells. Clonal analysis demonstrates that piglet NSPs are multi-potent and retain the capacity to generate both glia and neurons. These cells expressed VEGF, VEGFR1, VEGFR2 and Neuropilin-1 and -2 mRNAs. Real time PCR revealed that SVZ NSPs from newborn piglet expressed total VEGF and all VEGF splice variants. These findings show that piglet NSPs may be helpful to more effectively design growth factor based strategies to enhance endogenous precursor cells for cell transplantation studies potentially leading to the application of this strategy in the nervous system disease and injury.

Use of Aspiration Method for Collecting Brain Samples for Rabies Diagnosis in Small Wild Animals

P>In developing countries such as Brazil, where canine rabies is still a considerable problem, samples from wildlife species are infrequently collected and submitted for screening for rabies. A collaborative study was established involving environmental biologists and veterinarians for rabies epidemiological research in a specific ecological area located at the São Paulo State, Brazil. The wild animals' brains are required to be collected without skull damage because the skull's measurements are important in the identification of the captured animal species. For this purpose, samples from bats and small mammals were collected using an aspiration method by inserting a plastic pipette into the brain through the magnum foramen. While there is a progressive increase in the use of the plastic pipette technique in various studies undertaken, it is also appreciated that this method could foster collaborative research between wildlife scientists and rabies epidemiologists thus improving rabies surveillance.

Gene expression in placentation of farm animals: An overview of gene function during development

Eutherian mammals share a common ancestor that evolved into two main placental types, i.e., hemotrophic (e.g., human and mouse) and histiotrophic (e.g., farm animals), which differ in invasiveness. Pregnancies initiated with assisted reproductive techniques (ART) in farm animals are at increased risk of failure; these losses were associated with placental defects, perhaps due to altered gene expression. Developmentally regulated genes in the placenta seem highly phylogenetically conserved, whereas those expressed later in pregnancy are more species-specific. To elucidate differences between hemotrophic and epitheliochorial placentae, gene expression data were compiled from microarray studies of bovine placental tissues at various stages of pregnancy. Moreover, an in silico subtractive library was constructed based on homology of bovine genes to the database of zebrafish - a nonplacental vertebrate. In addition, the list of placental preferentially expressed genes for the human and mouse were collected using bioinformatics tools (Tissue-specific Gene Expression and Regulation [TiGER] - for humans, and tissue-specific genes database (TiSGeD) - for mice and humans). Humans, mice, and cattle shared 93 genes expressed in their placentae. Most of these were related to immune function (based on analysis of gene ontology). Cattle and women shared expression of 23 genes, mostly related to hormonal activity, whereas mice and women shared 16 genes (primarily sexual differentiation and glycoprotein biology). Because the number of genes expressed by the placentae of both cattle and mice were similar (based on cluster analysis), we concluded that both cattle and mice were suitable models to study the biology of the human placenta. (C) 2011 Elsevier B.V. All rights reserved.

Effects of intrauterine malnutrition on brain free amino acids of young rats, after nutritional recovery during lactation period

The effect of protein-calorie malnutrition during gestation on the brain amino acids of rat pups was studied following nutritional recovery during lactation. The brain amino acids of rat pups born to dam rats malnourished during gestation were studied after these rat pups received proper nutrition during lactation. Pregnant rats were fed a 1% protein diet with total caloric intake restricted to half that of controls. After birth, the offspring of rats fed on deficient diets were nurtured up to the 28th day postpartum by foster mothers receiving adequate diets. At this time, the offspring were killed. The control group consisted of offspring from pregnant rats fed a diet with adequate protein (21%) and calories during the entire gestation and lactation period. Quantitation of brain amino acids in the pups at 28 days postpartum showed lower concentrations of essential and nonessential amino acids in the rats malnourished during gestation. Concentrations of histidine, glycine, and α-aminobutyric acids were all reduced. These findings demonstrate that the brains of rat pups malnourished during gestation show persistent decreases in specific brain amino acids after adequate postpartum nutrition.

Does the male reproductive performance depend on the early lactation milk in rats?

This study investigated the role of the early lactation milk (ELM) on the male reproductive performance and the participation of GnRH on this effect in Wistar rats. Newborn males were divided into three experimental groups: 1) ELM-deprived pups, 2) ELM-deprived pups treated with exogenous GnRH 1, 7, 25, and 31 h after birth, and 3) non-ELM-deprived and without exogenous GnRH. In relation to the other two groups, the ELM-deprived male rats exhibited as adults: reduced fertility, decreased weight of both vas deferens and seminal vesicle, and reduced levels of fructose in the seminal vesicle and prostate gland. We suggest that the intake of ELM during the neonatal period is important to the later sexual development of rats, and that GnRH is somehow involved in such an effect.

Long-term effect of prolactin treatment on glucose-induced insulin secretion in cultured neonatal rat islets

Insulin secretion and 45SCa2+ uptake and efflux were studied in neonatal rat islets maintained in culture for 7 or 19 days in the absence or presence of prolactin (PRL). Insulin secretion in response to glucose (G), leucine (Leu), arginine (Arg) and carbachol (Cch) was augmented after 7 and 19 days in culture, compared to basal secretion (G 2.8 mM), in both PRL- treated and control islets. However, the increase in insulin secretion induced by the above secretagogues was higher in islets cultured in the presence of PRL for 19 days. In PRL-treated islets, the 45Ca2+ content after a 5 min incubation in the presence of G, Leu, Arg and Cch was significantly higher than the control only in islets cultured for 19 days. Except with Arg, the 45Ca2+ uptake in PRL-treated islets after a 90 min incubation was also significantly higher than the control only in islets cultured for 19 days. Finally, Leu-induced alterations in the 45Ca2+ efflux were higher in PRL-treated than in control islets cultured for 7 or 19 days. In the absence of external Ca2+, the reduction in 45Ca2+ efflux induced by glucose was also significantly higher in PRL-treated than in control islets. This effect was slightly potentiated after 19 days in culture. These data further support the hypothesis that PRL treatment enhances maturation of the secretory mechanism in neonatal islets. This effect can be potentiated even more if the treatment is prolonged.

Ticks (Acari: Ixodidae) associated with wild animals in the Pantanal region of Brazil

This paper describes the identification of ticks from wild animals of the Pantanal region in Brazil as part of a comprehensive study about established and emerging tick-host relationships and related pathological aspects. Eighty-one animals were captured (representing 13 species six orders), and ticks were found on 63 (78%). Tick species identified included Boophilus microplus (Canestrini), Amblyomma cajennense (F.), Amblyomma parvum Aragão, Amblyomma pseudoconcolor Aragão, Amblyomma scalpturatum Neumann, Amblyomma nodosum Neumann, Amblyomma ovale Koch, and Amblyomma tigrinum Koch. Dragging from grasslands yielded negative results compared with the high concentration of ticks that were collected from leaves in the forests.

Evaluation of lacrimal system outflow in the newborn by dye tests

PURPOSE: The aim of this study was to evaluate the outflow from the lacrimal excretory system (LES) at birth. METHOD: Two-hundred LES were evaluated by the Milder Test (DDT) and the modified Jones I Test (JIT) in 100 newborns (NBs) from 31 (T 1) until 42 (T 3) weeks of gestational age. The occurrence of obstruction was related to sex, weight at birth, presence of ocular discharge and the degree of maturity. RESULTS: Both tests were in agreement and showed LES obstruction in 61.5% (DDT) and 78.5% (JIT), respectively, while the LES outflow increased with increasing maturity. CONCLUSION: Evaluation of LES outflow at birth reveals obstruction rates that are much higher than those clinically observed in children with complaints of epiphora.

Ticks associated with wild animals in the Nhecolândia Pantanal, Brazil

A study of ticks associated with wild animals was carried out from September 1996 to April 1998 at the Fazenda Alegria (21,000 ha), in the Nhecolândia Pantanal, State of Mato Grosso do Sul, Brazil, a sunken plain bordering the upper Paraguay river, located 19 × 08′S; 56 × 46′W. A total of 81 wild animals (13 species, 6 orders) were captured with the aid of nets, and ticks were found on 63 (78%). Tick species identified included Boophilus microplus (Canestrini), Amblyomma cajennense (F.), A. parvum (Aragão), A. pseudo-concolor (Aragão), A. scalpturatum (Neumann), A. nodosum (Neumann), A. ovale (Koch), and A. tigrinum (Koch). Dragging from grasslands (campos) yielded negative results compared to the high concentration of ticks, mainly nymphs, that were collected from leaves in the forests (capão). Predominance of immature instars (Amblyomma genera) was observed in the end of winter (August-September). Ticks were associated mainly with coatis, deer (Mazama gouazoubira) and anteater, and these animals may play a role in the epidemiology of tick-transmitted pathogens in the Pantanal if one considers their coexistence with local domestic animals.

A review of antifertility folkloric plants tested in laboratory animals

The use of natural active principals is widespread among a great proportion of the rural population, or by people who do not have easy access to medical assistance. These active principles are used as food or medicines, and even for purposes of contraception. It becomes necessary to establish a relationship between the folklore habits and current information on the nature of anti-fertility substances, and knowledge of their mechanisms. Anti-fertility agents may exert their actions in a number of areas, (hypothalamus, anterior pituitary, oviduct, uterus, and vagina), inhibiting synthesis and/or liberation of hormones (follicle-stimulating, luteinizing, and steroid hormones), ovulation, ovum transportation, and implantation process. Therefore, a review of literature was carried out, including of several plants used by women as abortifacient and anti-fertility agents to compare their effects with those obtained among laboratory animals.

Neonatal inhalatory anesthetic exposure: Reproductive changes in male rats

We investigated the effects of an inhalatory anesthetic (ethyl ether) during the neonatal period of brain sexual differentiation on the later fertility and sexual behavior of male rats. Animals were exposed to ethyl ether immediately after birth. At adulthood, body weight, testes wet weight, and plasma testosterone levels were not affected; however, neonatal exposure to ether showed alterations on male fertility: a decrease in the number of spermatids and spermatozoa, an increase in the transit time of cauda epididymal spermatozoa and a decrease in daily sperm production. An alteration of sexual behavior was also observed: decreased male sexual behavior and appearance of homosexual behavior when the male rats were castrated and pretreated with exogenous estrogen. Probably, the ether delayed or reduced the testosterone peak of the sexual differentiation period, altering the processes of masculinization and defeminization of the hypothalamus. Our results indicate that perinatal exposure to ethyl ether during the critical period of male brain sexual differentiation, acting as endocrine disruptors, has a long-term effect on the fertility and sexual behavior of male rats, suggesting endocrine disruption through incomplete masculinization and defeminization of the central nervous system. © 2002 Elsevier B.V. All rights reserved.

Neuroendocrine alterations impair enamel mineralization, tooth eruption and saliva in rats.

Neonatal administration of monosodium glutamate (MSG) in rats causes definite neuroendocrine disturbances which lead to alterations in many organ systems. The possibility that MSG could affect tooth and salivary gland physiology was examined in this paper. Male and female pups were injected subcutaneously with MSG (4 mg/g BW) once a day at the 2nd, 4th, 6th, 8th and 10th day after birth. Control animals were injected with saline, following the same schedule. Lower incisor eruption was determined between the 4th and the 10th postnatal days, and the eruption rate was measured between the 43rd and the 67th days of age. Pilocarpine-stimulated salivary flow was measured at 3 months of age; protein and amylase contents were thereby determined. The animals treated with MSG showed significant reductions in the salivary flow (males, -27%; females, -40%) and in the weight of submandibular glands (about -12%). Body weight reduction was only about 7% for males, and did not vary in females. Saliva of MSG-treated rats had increased concentrations of total proteins and amylase activity. The eruption of lower incisors occurred earlier in MSG-treated rats than in the control group, but on the other hand the eruption rate was significantly slowed down. The incisor microhardness was found to be lower than that of control rats. Our results show that neonatal MSG treatment causes well-defined oral disturbances in adulthood in rats, including salivary flow reduction, which coexisted with unaltered protein synthesis, and disturbances of dental mineralization and eruption. These data support the view that some MSG-sensitive hypothalamic nuclei have an important modulatory effect on the factors which determine caries susceptibility.

Static lung compliance and body pressures in Tupinambis merianae with and without post-hepatic septum

The surgical removal of the post-hepatic septum (PHS) in the tegu lizard, Tupinambis merianae, significantly reduces resting lung volume (VLr) and maximal lung volume (VLm) when compared with tegus with intact PHS. Standardised for body mass (MB), static lung compliance was significantly less in tegus without PHS. Pleural and abdominal pressures followed, like ventilation, a biphasic pattern. In general, pressures increased during expiration and decreased during inspiration. However, during expiration pressure changes showed a marked intra- and interindividual variation. The removal of the PHS resulted in a lower cranio-caudal intracoelomic pressure differential, but had no effect on the general pattern of pressure changes accompanying ventilation. These results show that a perforated PHS that lacks striated muscle has significant influence on static breathing mechanics in Tupinambis and by analogy provides valuable insight into similar processes that led to the evolution of the mammalian diaphragm. © 2003 Elsevier Science B.V. All rights reserved.