Toxoplasma gondii (T. gondii) is the causative agent of toxoplasmosis. This protozoan has the characteristic of being obligate intracellular and having a high prevalence worldwide, where it is believed to have infected one third of the world's population, causing great morbidity and mortality. Given the complexity of this disease, several studies have been dedicated to the study of structures that are associated with parasitic diseases. Among these structures are the Communicating Junctions, which are responsible for the exchange of ions and small messengers that maintain tissue homeostasis. These transmembrane channels play an important role in intercellular communication in different tissues, as they allow communication in different cell types, including macrophages. With this, the morphological and functional characterization of gap junctions in macrophages, and in particular formed by connexin 43, has been the subject of study by several groups, but their regulatory mechanisms still deserve clarification, especially in the face of pathological changes, such as in infectiousinflammatory processes caused by T. gondii. In view of this, the main objective of this study was to evaluate the modulation of gap junctions in the macrophage lineage J774- G8, after infection with the parasite Toxoplasma gondii and subsequent activation with inflammatory pro-immune factors. As methodology, it was used: (1) Culture of J774-G8 cells; (3) Infection of the culture by the T. gondii strain RH; (4) Treatment with individual and conjugated pro-immunoinflammatory factors (IFN-γ, TNF-α, IFN-γ + TNF-α); (5) Immunoelectrophoretic assays (Western Blot); and (4) Immunofluorescence assays and analysis by confocal microscopy. The general results found were: (1) Improvement in the morphological profile of cultures of J774-G8 cells infected with T. gondii treated with proimmune-inflammatory factors; (2) Increased Cx43 protein expression in infected J774- G8 cells after treatment with pro-inflammatory immune factors for 24 and 48 hours; (3) Cell activation stimulated by treatment with conjugated factors; (4) The damage to the cellular cytoskeleton caused by the infection was irreversible, even after treatment with inflammatory pro-immune factors in infected cells; (5) Damage to the cytoskeleton prevented the transport and anchoring of Cx43 in the plasmatic membrane, however the factors provided an increase in the cytoplasmic levels of Cx43. With this, it was possible to conclude that: infection with T. gondii causes irreversible damage to macrophage cells, however treatment with pro-immune inflammatory factors stimulates the production of Cx43, which even though it fails to insert itself into the plasmatic membrane in infected cells due to damage to the cytoskeleton, may play important roles in the process of maintaining the infected cell structure.

Analgesics are among one of the most commonly used drug classes for the treatment or relief of painful and inflammatory conditions. However, due to the large number of side effects and often the ineffectiveness of these drugs, it is necessary to search for new drugs. When a new compound shows promising responses, the conjugation of specific bioactive structures in a single molecule can be performed through molecular hybridization. An effective way to rationally plan new drugs that have synergistic action and/or decrease adverse effects. The aim of this study was to evaluate the acute and chronic antinociceptive and anti-inflammatory activity of the new hybrid compound: (±-cis) (6-ethyl-tetrahydro-2H-pyran-2-yl)methyl 2-(2-(2, 6-dichlorophenylamino) phenyl acetate (LS26), synthesized from the hybridization of the compound [(±)-Cys)(6-ethyl-Tetrahydro-2H-Pyran-2-yl]Methanol (LS20) and the non-inflammatory anti-inflammatory steroid diclofenac sodium. The compound LS26 was administered orally in all animals and in all experimental models. According to the results, the compound has acute antinociceptive activity observed in the abdominal writhing models, formalin (both phases) and immersion of the tail in hot water. The mechanism of action of the compound involves the opioid system, considering that the application of selective opioid antagonists were able to reduce the antinociceptive effect of the hybrid compound. The NO/cGMP/K+ATP pathway is also one of the mechanisms of action of LS26 due to the reduction of the antinociceptive effect with the administration prior L-NAME, ODQ and glibenclamide. Acute and chronic antinociceptive activity on allodynia induced by chronic constriction of the sciatic nerve was also observed in the neuropathic pain model due to inhibition of IL-1β and IL-6 production in the sciatic nerve. The compound shows no relationship between the antinociceptive effect and a motor deficit, according to the results of the rota-rod test. As for anti-inflammatory activity, the LS26 compound showed anti-edematogenic activity, as it was able to reduce leukocyte migration and the production of pro-inflammatory cytokines such as IL-1β, TNF-α and IL-6 and increase cytokine synthesis anti-inflammatory IL-10 in the subcutaneous air bag model. The compound showed selectivity for COX-2 inhibition observed in in vitro evaluation. In the toxicological test, the compound did not present acute and subchronic alterations in the microscopic and macroscopic evaluations. These results indicate antinociceptive activity through opioid receptors and the NO/cGMP/K+ATP pathway, in addition to anti-inflammatory activity with inhibition of leukocyte migration, reduction of IL-1β, TNF-α and IL-6 levels, increase of IL-10 and selective inhibitory activity on COX-2.

Introduction: The present work was carried out with the support of the Higher Education Personnel Improvement Coordination – Brazil (CAPES) - Finacing code 001. The spontaneously hypertensive animal strain (SHR) is considered as a model of essencial hypertension, being the model that is closer to the hypertension of the human being. This strain up to 4 weeks of age does not present hypertention, which makes it interesting to study phenotipic changes that occur during this animals’ life span. Males and females are diferent in many physiological aspects. The body sodium reserve can be stored in several tissues, being the skin reserve, the interest of this study and in the SHR lineage this aspect has not been elucidated yet. Objectives: To evaluate the physiological status of active and inactive reserves; evaluate changes in the animals’s ingestive pattern making a relationship with blood pressure and sodium reserve; to evaluate diferences between males and females in relation to ingestive behavior; to evaluate changes in hormone concentrations before and after the establishment of hypertention in 4 weeks and 12 weeks old animals. Material and methods: Male and female SHR and Wistar animals, 4 weeks and 12 weeks old, were used. The animals were obtained from the Central Biottery of the Department of Physiological Sciences of UFRRJ. They were kept under standard conditions of light, temperature, and food. Experiments were performed in metabolic cages, radioimmunoassay, tail cuff for pressure assessments and skin sodium and water content analizes. Results: Protocol 1. At the age of 4 weeks SHR males take more sodium from 26 days of age and females from 30 days. Protocol 2: SHR lineage animals had lower weight indexes than Wistar animals. There were no lineage differences in the hormones AVP, OT and ANG II at 4 weeks and 12 weeks of age in females, males SHR and Wistar at 4 weeks of age were no different. Protocol 3: Spontaneously hypertensive animals (SHR) are not hypertensive at 4 weeks of age and become hypertensive at 5 weeks of life. Protocol 4. Accumulated active or inactive reserve does not change with lineage in adult males or offspring, but increases with advancing age in males and females. Discussion: Protocol 1: As expected SHR animals take more hypertonic saline than male or female Wistar animals. Protocol 2: Hormonal differences in AVP are due to physiological variations of gonadal hormones and increased plasma concentration of ANG II in SHR was expected as a trait of the strain. Protocol 3: The increase in the mean arterial pressure was observed in the 5^th week of life for the male SHR rats and in the 6^th week of life for the female SHR. Protocol 4. In males, hypertension does not make the accumulated sodium reserve different from normotensive animals. Conclusions: Protocol 1: Sodium and water intake are influenced by lineage and sexual dimorphism. Protocol 2: Most hormonal variations occurred due to lineage differences. Protocol 3: The essencial hypertension is related to the age of the animals. Protocol 4. Hypertension does not change osmotically active or inactive reserve under physiological conditions, but age and sexual dimorfism play importante roles.

Inadequate maternal nutrition is related to the development of diseases and health problems, such as obesity and overweight, cardiovascular diseases and can cause metabolic programming in your offspring. Our research group previously demonstrated that maternal consumption of a high-fat diet promoted an increase in body mass, adiposity, hyperleptinemia in the offspring of rats at weaning, in addition to impaired systolic function at 30 days of age. This study aimed to investigate the effect of maternal consumption of a high-fat diet on the blood pressure of adult rats and the possible involvement of sodium ion in this process. For this, Wistar rats received a control diet (9% lipids, group C) or high-fat diet (29% lipids, group HFD ) for 8 weeks before mating, and during pregnancy and lactation. The progenitors C and DH gave rise to the control (C) and hyperlipidic (HFD) offspring, respectively. After weaning, part of the offspring was euthanized and the rest at 180 days of age. At weaning were evaluated: body weight, cardiac hypertrophy, weight of kidneys and white adipose tissues, sodium concentration in blood and skin and at 180 days were evaluated: body weight, food and water intake, kidney weight, indirect assessment of cardiac hypertrophy , white adipose tissues, sodium concentration in the blood, urine and skin. In urine in addition to urinary volume, creatinine, Na⁺, K⁺, Cl^-, normalized were also analyzed. body weight, urine volume and water intake were measured and urine samples were collected. Upon weaning, the offspring showed greater body mass, cardiac hypertrophy, increased dry skin weight and total skin water content was reduced, as well as the relative sodium of the skin. At 180 days of age, there was no variation in body mass, white adipose tissue, water and food intake, kidney weight as well as in the indirect assessment of cardiac hypertrophy. The HFD males showed higher systolic pressure compared to the C males as well as an increase in plasma sodium, with no change in urinary sodium between the groups, in biochemical analyzes only in the HFD males a decrease in the Cl- electrolyte was observed. The total water content of the skin was reduced in HFD males. None of these changes were detected in the females of the groups. When assessing sodium concentrations in urine and epithelial samples, there was no difference. Our results demonstrate that maternal consumption of a high-fat diet promotes an increase in plasma sodium concentration and systolic pressure in adult offspring in a sex-specific way. These events may be related to the development of cardiovascular diseases, such as hypertension.

Anxiety disorders and depression are among the most prevalent mental disorders in the population, affecting thousands of people around the world. Folk medicine, a very old practice, makes use of several plants for the purpose of treating diseases that affect man. Over the years there has been a great advance in the studies of medicinal plants to obtain new compounds with therapeutic properties. Schinus molle L., plant belonging to the family Anacardiaceae, is popularly known as aroeira. In addition to presenting a wide range of biological activities, its ethanolic and hexanic extract showed to have antidepressant effect. The objective of this project was to evaluate the anxiolytic and antidepressant actions of the essential oil of the leaves of Schinus molle L. (OEFS) in animal models. In the open field  test (AC), OEFS at a concentration of 100 mg/kg reduced the number of quadrants traveled (OES100 = 19.1 ± 6.2 vs. CT = 71.4 ± 3.3) as well as the number of rearings (OES100 = 1 ± 0.5 vs. CT = 22.5 ± 10.5). In the test of the rotating bar, the OESF did not present significant difference in the time of permanence in the bar and number of falls. In the test of sleep induced by etomidate, OESF100 showed an increase in total sleep recovery time in relation to the control group (OES100 = 162.2 ± 14.9 vs. CT = 97.4 ± 7.3 s). To evaluate the possible anxiolytic effect, the doses of 25 and 50 mg / kg OEFS were used in the elevated plus maze (LCE) and in the MARBLE BURYING test (TEE). In LCE, OEFS did not present significant difference in any of the parameters analyzed (percentage of time in open arms and percentage of open arms entries). In the TEE, OEFS did not reduce the number of Glass marbles burying in relation to the control group. For the evaluation of a possible like-antidepressant effect of OEFS, the forced swimming test (NF) was performed. OEFS did not reduce the immobility time at any of the doses tested (25, and 50mg/kg). Pretreatment with flumazenil was not able to reverse the reduction of locomotor and exploratory activity of the animal in CA caused by OEFS100, showing that the depressant effect is not mediated by the benzodiazepine site of the GABAA receptor. Further studies are needed for further investigation of its sedative effect, as well as better exploration of the pharmacological profile of this plant.

The decrease of the body fluids can be caused by water deprivation and, therefore, control and regulation mechanisms are activated in order to ensure the maintenance of hydroelectrolytic balance, among them the secretion of neurohypophyseal hormones. Water deprivation in birds is an important stress stimulus that causes an increase in the activity of NPV and NSO neurons, called magnocellular cells, and consequently an increase in water intake behavior in birds. Thus, water deprivation becomes a convenient experimental model for studies of vasotocinergic and mesotocinergic activity, as well as studies of the interaction of these peptides with their respective receptors and their central and systemic action. Thus, the present study investigated the possible correlations between vasotocinergic and mesotocinergic activity in birds with behavioral changes between thirst mechanism and water intake, through an experimental model developed with Japanese quails (Coturnix coturnix japonica), submitting the experimental groups an water deprivation. Male quails of approximately 50 days were used, maintained at a controlled temperature of 26 ± 2ºC, under a photoperiod of 12/12 hours, subdivided into 3 distinct groups (n = 6): Control Group (CTRL), Water Deprivation for 36 hours (PvH) and Water Deprivation for 36 hours with 2 hours of water presentation (IH-2h). We found that water deprivation was able to increase the water intake of the IH-2h group for all observed time intervals when compared to the CTRL group and the IH-2 group before deprivation (IH-2h after deprivation vs CTRL, p IH-2h after deprivation vs IH-2h before deprivation, p <0.0001). The PvH group showed a statistically significant difference when compared to the CTRL group in basal water intake at 120 minutes (PvH 3.56 ± 3.32 vs CTRL 0.79 ± 0.70; p = 0.002). There was a significant reduction in body weight in the PvH and IH-2h groups when considering the variation in weight gain before and after deprivation (PvH after deprivation vs CTRL, p <0.0001 and IH-2h after deprivation vs CTRL, p <0.0001). Thus, the present study was able to demonstrate positive results regarding water ingestive behavior and its possible influences on species body weight gain after water deprivation. Although the results of the plasma parameters evaluated in this study do not allow us to confirm that the vasotocinergic and mesotocinergic neuroendocrine control mechanisms are involved in water deprivation, but we suggest that the action of the hormones AVT and MT is extremely important in the body fluids control. Further studies are needed to elucidate the role of these and other hormones involved in the osmoregulation of Japanese quails.

Although classically involved in cardiovascular control and hydromineral balance, the renin-angiotensin system (SRA) has been implicated in energy balance control. Angiotensin II (ANG II) stimulating its type 1 receptor (AT1) may change body weight and food ingestion in some experimental models of obesity. One of the main sites of action of circulating ANG II is the subfornical organ (SFO), a circumventricular organ related to the control of several autonomic functions, motivated behaviors, and energetic metabolism. Thus, our hypothesis is that ANG II may act on AT1 receptors to promote an integrate control of metabolic and hydroelectrolytic balance during challenges of energetic homeostasis. This thesis aims three main goals: a) investigate if a same SFO neuron is sensitive to ANG II and glucose, and if glucose concentration may influence this cell’s response to ANG II; b) evaluate if food deprivation, as a hypoglycemic stimulus, may alter systemic SRA activity and AT1 expression at SFO; c) and evaluate if ANG II through AT1 may influence food and/or water ingestion after food deprivation. Male Wistar and Sprague-Dawley male rats were used and all procedures have been approved by ethics committees in experimental animal use, respectively: Queen’s University 2017-1735 and FMRP-USP 229/18. SFO neurons that responded to ANG II may be found in different phenotype populations of glucosensing, either non-glucosensitive (43%), glucoexcited (52%) or glucoinhibited (71%). Cultures of SFO cells grown at different glucose concentrations for 1, 6 or 24h did not change expression of AT1 receptors. Nevertheless, electrophysiological studies, shows that approximately 31% (n=4/13) of tested SFO neurons didn’t responded to ANG II at 5mM glucose medium but depolarized to ANG II at 10mM. It suggests that metabolic state of the organism may change the way the central nervous system respond to ANG II. Food deprivation lead to and increased plasmatic concentration of ANG II and of mRNA for AT1a receptors at SFO. Treatment with losartan (an AT1 blocker) at low doses in drinking water, for 48h fasting attenuated the glycemic drop. After access to food was reestablished, there’s only a tendency of decrease of food ingestion at 60 min in the group treated with losartan. By that same one-hour period, this same group exhibited a significative reduction in water intake. Yet, after 12 hours of intracerebroventricular (i.c.v.) microinjections of losartan lead to food and water ingestion decrease in basal condition, while after a 48h fasting, only water ingestion was significantly decreased 12h after losartan i.c.v. Though we might conclude that SRA is activated in fasting conditions, followed to an increase in AT1 expression at SFO, which is related to water ingestion during feeding, as an important mechanism of maintenance of volume and osmolality of extracellular fluid during food ingestion.

Thirst is an essential sensation for the development behavioral changes that driving the
motivation for drink water, a vital response for the maintenance of hydrosaline
homeostasis. Animals that are submitted to water or food deprivation in general are
more willing to take risks for their survival, in this sense, water deprivation and salt
loading were stimuli used to trigger intracellular thirst. Under these conditions
vasopressin (AVP) and oxytocin (OT) are secreted by magnocellular neurons through
the neurohypophysis and release for the systemic circulation. Namely, recent studies
have demonstrated that vasopressinergic magnocellular neurons project to
extrahypothalamic regions, which include the habenula and amygdala. Thus, the present
study aimed to analyze the effect of increased plasma osmolality on exploratory and
anxiety-like behaviors in rats subjected to water deprivation and salt loading, as well as
evaluating the gene expression of type 1a AVP (AVPR1a) and OT (OTR) receptors in
the basolateral amygdala, central amygdala and habenula, as well as the enzyme
synthesizing GABA (GAD67) and the glutamate vesicular transporter (VGLUT2) in
both nuclei. Male Wistar rats were randomly divided into five experimental groups:
Control (CT), 24 hour water deprivation (PH24h) and 48 hours (PH48h) and salt
loading with 1.8% NaCl for 24 hours (SS24h) and 48 hours (SS48h). All groups were
submitted to plasma analysis, behavioral tests and analysis of gene expression through
RT-qPCR. All data are described as mean ± standard error of the meanand the
differences were considered significant when p < 0.05. Plasma osmolality increased
significantly in the PH48h (p = 0.0039) and SS48h (p = 0.0176) groups. In the elevated
plus maze, there was increase in the percentage of open arm entries in groups PH48 (p =
0.0079) and SS48h (p = 0.0015), as well as the time spent in open arms in PH48h (p =
0.0029) and SS48h (p <0.0001), indicating an anxiolytic response. In addition, linear
regression was significant between plasma osmolality vs time spent in the open arms (p
= 0.0004, r = 0,259). In the open field no difference was observed, indicating that the
animals did not increase the spontaneous locomotor activity. In the light and dark box,
only the PH48h group (p = 0.0424) showed an increase in time spent in the light area. In
relation to gene expression, we observed a significant increase in the expression of
AVPR1a (p = 0.0345) and OTR (p = 0.0103) in the PH48h group in the central
amygdala. Our results indicate that plasma hyperosmolarity induced by 48 hours of
water deprivation or salt loading is capable of eliciting anxiolytic behaviors in rats and
it’s possible that AVPR1a and OTR receptors mediate this response in the central
amygdala.