Behavioral, neurochemical and neuroimmune features of RasGEF1b deficient mice.

The factor RasGEF1b is a Ras guanine exchange factor involved in immune responses. Studies have also implicated RasGEF1b in the CNS development. It is still limited the understanding of the role of RasGEF1b in CNS functioning. Using RasGEF1b deficient mice (RasGEF1b-cKO), we investigated the impact of this gene deletion in behavior, cognition, brain neurochemistry and microglia morphology. We showed that RasGEF1b-cKO mice display spontaneous hyperlocomotion and anhedonia. RasGEF1b-cKO mice also exhibited compulsive-like behavior that was restored after acute treatment with the selective serotonin reuptake inhibitor (SSRI) fluoxetine (5 mg/kg). A down-regulation of mRNA of dopamine receptor (Drd1, Drd2, Drd4 and Drd5) and serotonin receptor genes (5Htr1a, 5Htr1b and 5Htr1d) was observed in hippocampus of RasGEF1b-cKO mice. These mice also had reduction of Drd1 and Drd2 in prefrontal cortex and 5Htr1d in striatum. In addition, morphological alterations were observed in RasGEF1b deficient microglia along with decreased levels of hippocampal BDNF. We provided original evidence that the deletion of RasGEF1b leads to unique behavioral features, implicating this factor in CNS functioning.

Weight-drop model as a valuable tool to study potential neurobiological processes underlying behavioral and cognitive changes secondary to mild traumatic brain injury.

The pathophysiology of post-traumatic brain injury (TBI) behavioral and cognitive changes is not fully understood, especially in its mild presentation. We designed a weight drop TBI model in mice to investigate the role of neuroinflammation in behavioral and cognitive sequelae following mild TBI. C57BL/6 mice displayed depressive-like behavior at 72 h after mild TBI compared with controls, as indicated by a decrease in the latency to first immobility and climbing time in the forced swim test. Additionally, anxiety-like behavior and hippocampal-associated spatial learning and memory impairment were found in the elevated plus maze and in the Barnes maze, respectively. Levels of a set of inflammatory mediators and neurotrophic factors were analyzed at 6 h, 24 h, 72 h, and 30 days after injury in ipsilateral and contralateral hemispheres of the prefrontal cortex and hippocampus. Principal components analysis revealed two principal components (PC), which represented 59.1% of data variability. PC1 (cytokines and chemokines) expression varied between both hemispheres, while PC2 (neurotrophic factors) expression varied only across the investigated brain areas. Our model reproduces mild TBI-associated clinical signs and pathological features and might be a valuable tool to broaden the knowledge regarding mild TBI pathophysiology as well as to test potential therapeutic targets.

Innate and adaptive immune gene expression in the brain is associated with neuropathological changes after infection with bovine alpha-herpesvirus-5 in mice.

Bovine alpha herpesvirus-5 (BoAHV-5) is related to the development of meningoencephalitis in cattle. Very little is known about the molecular pathways involved in the central nervous system (CNS) damage associated with inflammation during BoHV-5 infection in mice. To better identify the specific immunological pathways triggered by BoAHV-5 infection in mice, we evaluated the mRNA expression of 84 genes involved in innate and adaptive immune responses. We compared gene expression changes in the cerebrum from noninfected and infected mice with BoHV-5 at a 1 × 107 TCID50. Then, we analyzed the association of these genes with neurological signs, neuropathology, and activation of glial cells in response to BoHV-5 infection. Three days after BoAHV-5 infection, increased expression of TNF, IL-2, CXCL10, CXCR3, CCR4, CCL5, IFN-γ, IL-10, IRF7, STAT1, MX1, GATA 3 C3, LIZ2, caspase-1 and IL-1b was found. We also observed the upregulated expression of the CD8a, TBX21 and CD40LG genes and the downregulated expression of the CD4 gene after BoAHV-5 infection. In addition, BoHV-5-infected animals showed higher levels of all the evaluated inflammatory mediators (TNF, IFN-γ and IL-10) on day 3 postinfection. BoAHV-5-infected animals showed neurological changes along with meningoencephalitis, neuropil vacuolation, hemorrhage and reactive gliosis. Astrogliosis and microgliosis, indicated by increased expression of glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (Iba-1), were found throughout the neuropil in infected brains. Moreover, cleaved caspase-3 immunopositive glio-inflammatory cells were visualized around some blood vessels in areas of neuroinflammation in the cerebrum. In agreement on that we found higher cleaved caspase-3 and Iba-1 expression evaluated by western blot analysis in the brains of infected mice compared to control mice. In conclusion, our results revealed.

Role of cytokine and neurotrophic factors in nicotine addiction in the conditioned place preference paradigm.

The mechanisms involved in the maintenance of cigarette smoking and nicotine reward remain unclear. Immune response might play an important role in this context. Nicotine may induce both central and systemic inflammatory responses as well as changes in the regulation of brain-derived neurotrophic factor (BDNF). The conditioned place preference (CPP) is a method used for the evaluation of nicotine-induced reward, reproducing nicotine-seeking behavior in humans. So far, there are no studies investigating the relationship between neuroinflammation and nicotine-induced CPP. This study aimed to evaluate the levels of inflammatory mediators and neurotrophic factors in key areas of the central nervous system (CNS) of mice subject to nicotine-induced CPP. CPP was induced with an intraperitoneal administration of 0.5 mg/kg of nicotine in male Swiss mice, using an unbiased protocol. Control group received vehicle by the same route. The levels of cytokines, chemokines, and neurotrophic factors were measured using Enzyme-Linked Immunosorbent Assay (ELISA) in the brain after CPP test. As expected, nicotine induced place preference behavior. In parallel, we observed increased peripheral levels of IL-6 and IL-10 alongside increased hippocampal levels of NGF but decreased GDNF in mice treated with nicotine compared to controls. In the striatum, nicotine promoted decrease of IL-1ß, IL-10 and GDNF levels, while the levels of all the mediators were similar between groups in the pre-frontal cortex. Our results provide evidence on the role of cytokines and neurotrophic factors in nicotine-induced CPP in mice.

Brain-derived neurotrophic factor is down regulated after bovine alpha-herpesvirus 5 infection in both wild-type and TLR3/7/9 deficient mice.

Neurotrophic factors have been implicated in the control of neuronal survival and plasticity in different brain diseases. Meningoencephalitis caused by bovine alpha-herpesvirus 5 (BoHV-5) infection is a frequent neurological disease of young cattle, being the involvement of apoptosis in the development of neuropathological changes frequently discussed in the literature. It's well known that Toll-like receptors (TLRs) can activate neuroinflammatory response and consequently lead to neuronal loss. However, there are no studies evaluating the expression of neurotrophic factors and their association with brain pathology and TLRs during the infection by BoHV-5. The current study aimed to analyze brain levels of neurotrophic factors along with neuropathological changes during acute infection by BoHV-5 in wild-type (WT) and TLR3/7/9 (TLR3/7/9-/-) deficiency mice. The infection was induced by intracranial inoculation of 1 × 104 TCID50 of BoHV-5. Infected animals presented similar degrees of clinical signs and neuropathological changes. Both infected groups had meningoencephalitis and neuronal damage in CA regions from hippocampus. BoHV-5 infection promoted the proliferation of Iba-1 positive cells throughout the neuropil, mainly located in the frontal cortex. Moreover, significant lower levels of brain-derived neurotrophic factor (BDNF) were detected in both BoHV-5 infected WT and TLR3/7/9 deficient mice, compared with non-infected animals. Our study showed that BDNF down regulation was associated with brain inflammation, reactive microgliosis and neuronal loss after bovine alpha-herpesvirus 5 infection in mice. Moreover, we demonstrated that combined TLR3/7/9 deficiency does not alter those parameters.

Minocycline treatment prevents depression and anxiety-like behaviors and promotes neuroprotection after experimental ischemic stroke.

Depression and anxiety have been reported as the major neuropsychiatric consequences following stroke. Minocycline, a neuroprotective drug has minimized depressive symptoms in patients with major depressive disorders and anxiety-like symptoms. In addition, minocycline demonstrated efficacy and seemed a promising neuroprotective agent in acute stroke patients. The present studied evaluated the effects of minocycline treatment on the depression and anxiety-like behaviors, brain damage and expression of inflammatory and neuroprotective mediators after transient global cerebral ischemia in C57BL/6 mice. Brain ischemia was induced by bilateral occlusion of the common carotids (BCCAo) for 25 min and subsequent reperfusion. Sham and BCCAo animals received minocycline at a dose of 30 mg/kg by intraperitoneal injection during 14 days. The locomotor activity, depression and anxiety-like behaviors were assessed by open field, forced swim and elevated plus maze tests, respectively. Then, the brains were removed and processed to evaluate brain damage by histological and morphometric analysis, hippocampal neurodegeneration using Fluoro-Jade C histochemistry, microglial activity using iba-1 immunohistochemistry, brain levels of TNF, IFN-γ, IL-6, IL-10, IL-12p70 and CCL2 by CBA, CX3CL1 and BDNF by ELISA assays. The animals developed depression and anxiety-like behaviors post-stroke and minocycline treatment prevented those neurobehavioral changes. Moreover, minocycline-treated BCCAo animals showed less intense brain damage in the cerebral cortex, brainstem and cerebellum as well as significantly reduced hippocampal neurodegeneration. BCCAo groups exhibited up-regulation of some cytokines at day 14 after ischemia and brain levels of CX3CL1 and BDNF remained unaltered. Our data indicate that the depression and anxiety-like behavioral improvements promoted by minocycline treatment might be related to its neuroprotective effect after brain ischemia in mice.

Physical training improves exercise tolerance, cardiac function and promotes changes in neurotrophins levels in chagasic mice.

AIMS: To investigate the effects of moderate aerobic physical training on cardiac function and morphology as well as on the levels of glial cell-derived neurotrophic factor (GDNF), nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) of animals infected with the Y strain of Trypanosoma cruzi. MAIN METHODS: Twenty-eight male C57BL/6 mice were distributed into 4 groups: sedentary control (SC), trained control (TC), sedentary infected (CHC) and trained infected (CHT). The infection was performed by intraperitoneal injection of trypomastigote forms and the animals were adapted to treadmill in the week before the beginning of the training protocol, initiated 45 days post infection. Maximal exercise test (TEM) was performed at the baseline as well as at the end of the 4th, 8th and 12th weeks of training. At the end of the 12th week, all animals were evaluated for cardiac morphology and function by echocardiography. KEY FINDINGS: CHC group showed a larger area of right ventricle (RVA), increased end-systolic volume and reduction in ejection fraction (EF), stroke volume (SV), cardiac output (CO) and fractional area change (FAC). The training reduced the RVA and improved the FAC of chagasic animals. GDNF level was higher in TC and CHC groups compared to SC in heart and BDNF levels were higher in CHC compared to SC in heart and serum. SIGNIFICANCE: Physical training ameliorated the cardiac function of infected animals and promoted adjusts in BDNF and GDNF levels. These findings evidenced these neurotrophins as possible biomarkers of cardiac function responsive to exercise stimulus.

Inflammatory changes in peripheral organs in the BACHD murine model of Huntington's disease.

Huntington's disease (HD) is a neurodegenerative disease caused by a CAG repeat expansion in the gene encoding the huntingtin protein (HTT). This expansion leads to the formation of mutant huntingtin protein (mHTT) that is expressed in many body tissue cells. The mHTT interacts with several molecular pathways within different cell types, affecting the regulation of the immune system cells. It is still very limited the understanding of the immune changes in peripheral tissues in HD. Herein, we investigated the levels of inflammatory and regulatory cytokines in peripheral organs (i.e. kidney, heart, liver and spleen) of the 12-month-old BACHD model of HD. This robust murine model closely resembles the human disease. We found significant changes in cytokine levels in all organs analyzed. Increased levels of IL-6 were found in the kidney, while levels of IL-6 and IL-12p70 were increased in the heart of BACHD mice in comparison with wild-type (WT) animals. In the liver, we observed enhanced IL-12p70 and TNF-α levels. In the spleen, there was an increase in the levels of IL-4 and a decrease in the levels of IL-5 and IL-6 in BACHD compared to WT. Our findings provide the first evidence that the BACHD model also exhibits immune changes in peripheral organs, opening an avenue for the investigation of the potential role played by peripheral inflammatory response in HD. Further studies are needed to systematically address the mechanisms and pathways underlying immune signaling in peripheral organs in HD.

Interactions between local renin angiotensin system and nitric oxide in the brain of Trypanosoma cruzi infected rats.

Chagas' disease (CD) is a zoonosis caused by the protozoan Trypanosoma cruzi. Besides being an important cause of cardiomyopathy, central nervous system (CNS) manifestations have also been reported in CD. Renin-Angiotensin System (RAS) plays a pathophysiological role in several brain disorders such as cerebrovascular and neurodegenerative diseases. A link between RAS and nitric oxide (NO) pathways has been described in CNS. For instance, Angiotensin-(1-7) increases NO expression in the brain, which may, in turn, help to control parasite load in response to T. cruzi infection. Herein, we investigated the levels of RAS components in the brain cortex in acute T. cruzi infection and the effect of L-NAME administration, an inhibitor of the enzyme NO synthase, in CNS infection and in RAS molecules. Male Holtzman rats were inoculated intraperitoneally with T. cruzi Y strain and received L-NAME or tap water from one day before the infection until 13 days post infection (dpi). Parasitemia was evaluated on alternate days from day 3 post-infection until day 13 in both T. cruzi infected groups. Histopathological analysis of the brain cortex was also performed. Brain cortex was collected from non-infected (controls) and infected rats at 13 dpi for RAS components assessment. Infected rats receiving L-NAME presented higher parasitemia, brain parasitism and inflammation compared with non-treated infected animals. The administration of L-NAME significantly decreased the levels of Angiotensin I Converting Enzyme 2 (ACE2). In conclusion, we provided preliminary evidence of the interaction between RAS and NO during the acute phase of T. cruzi infection.

Conhecimento e sentimentos das mulheres climatéricas sobre a doença coronariana / Knowledge and feelings of climacteric women on coronary disease / Conocimiento y sentimientos de las mujeres en edad de climaterio sobre la enfermedad coronaria

Objetivo: estudo qualitativo com o objetivo de descrever o impacto da descoberta da doença coronariana no cotidiano das mulheres climatéricas. Método: utilizou- -se, para interpretação dos dados, a análise de conteúdo de Bardin. Resultados e discussão: foram identificadas quatro categorias: "o conhecimento da doença coronariana"; "mudança na alimentação"; "mudanças no trabalho"; "a insegurança e o medo da morte". As mulheres manifestam certo desconhecimento em definir a doença coronariana. Compreendem que é grave e impõe riscos, porém sentem dificuldades em defini-la ou explicá-la. Reconhecem a importância do tratamento preconizado, seguindo as recomendações terapêuticas como a mudança de hábitos de vida, a utilização da terapia medicamentosa e dos tratamentos invasivos, como a cirurgia de revascularização do miocárdio e a angioplastia. Conclusão: as principais mudanças no cotidiano das mulheres estão relacionadas à alimentação e às atividades laborais e domésticas. A perda da autonomia imposta pela doença ocasiona dependência e inutilidade. A insegurança e o medo também estão presentes associados a significações simbólicas acerca do inesperado e da morte. As mudanças cotidianas após a descoberta da doença coronariana causam impacto físico, emocional e social a essas mulheres, bastante prejudicial à sua saúde na recorrência de eventos coronarianos ou para uma condição mais grave e incapacitante da doença coronariana.(AU)

Objective: this is a qualitative study aimed at describing the impact of the discovery of the coronary disease in the daily life of climacteric women. Method: Bardin content analysis was used for data interpretation. Results and discussion: four categories were identified: "knowledge of the coronary disease"; "Change in food"; "Changes in work"; "Insecurity and fear of death". The women showed a lack of knowledge in defining the coronary disease. They understand that it is a serious and risky disease, but they find it difficult to define or explain it. They recognize the importance of the recommended treatment, following the therapeutic recommendations as to the change of habits of life, the use of the drug therapy and the invasive treatments, as the surgery of revascularization of the myocardium and the angioplasty. Conclusion: the main changes in the daily life of women are related to food and work and domestic activities. The loss of autonomy imposed by the disease causes dependence and uselessness. Insecurity and fear are also present associated with symbolic meanings of unexpected and death. The daily changes after the discovery of the coronary disease cause physical, emotional and social impact to these women, quite harmful to their health in the recurrence of coronary events or to a more serious and incapacitating condition of the coronary disease.(AU)

Objetivo: estudio cualitativo con miras a describir el impacto del descubrimiento de la enfermedad coronaria en el cotidiano de las mujeres en edad de climaterio. Método: la interpretación de datos se realizó según el análisis de Bardin. Resultados y discusión: se identificaron cuatro categorías: "conocimiento de la enfermedad coronaria"; " cambios en la alimentación"; "cambios en el trabajo"; " la inseguridad y el miedo a la muerte". Las mujeres manisfestaron desconocimiento para definir la enfermedad coronaria. Entienden que es grave y trae riesgos pero les resulta dificil definirla o explicarla. Reconocen la importancia del tratamiento, que sigue recomendaciones terapéuticas tales como el cambio de costumbres de vida, la terapia medicamentosa y los tratamientos invasivos, como la cirugía de revascularización del miocardio y la angioplastia. Conclusión: los principales cambios en el cotidiano de las mujeres están relacionados con la alimentación y las actividades laborales y domésticas. La pérdida de autonomía impuesta por la enfermedad causa dependencia e inutilidad. La inseguridad y el miedo también están presentes asociados a significados simbólicos sobre lo inesperado y la muerte. Los cambios cotidianos después del descubrimiento de la enfermedad coronaria causan impacto físico, emocional y social, bastante perjudicial a la salud en la recurrencia de eventos coronarios o para una condición más grave e incapacitante de la enfermedad coronaria.(AU)

Sintomas psicológicos em mulheres climatéricas cardiopatas / Síntomas psicológicos en mujeres en climaterio y cardiópatas / Psychological symptoms in climacteric women with heart disease

RESUMO Objetivo: investigar os sintomas climatéricos psicológicos em mulheres cardiopatas. Método: estudo quantitativo, realizado em Hospital Universitário no Nordeste do Brasil, no período de outubro de 2016 a janeiro de 2017. Foram entrevistadas individualmente 221 mulheres climatéricas cardiopatas atendidas no Ambulatório de Cardiologia. Foram realizadas análises da estatística descritiva, de associação com o teste não paramétrico de Qui-quadrado de independência (c2) e teste de Correlação de Spearman, com o auxílio do programa SPSS Statistics 20. Resultados: houve predomínio de sintomas climatéricos muito intensos como a angústia/ansiedade, 75 (33,9%); esgotamento físico e mental, 61 (27,6%); estado de ânimo depressivo, 54 (24,4%); nervosismo, 59 (26,7%); e insônia, 45 (20,4%). Houve associação significativa nos sintomas climatéricos psicológicos entre si e entre os sintomas psicológicos e a depressão. Conclusão: os sintomas climatéricos psicológicos parecem tornar as mulheres mais propensas a transtornos emocionais, agravada pela existência de uma doença crônica como a cardiopatia.

RESUMEN Objetivo: investigar los síntomas climatéricos psicológicos en mujeres cardiópatas. Método: estudio cuantitativo, realizado en Hospital Universitario en Nordeste de Brasil, en el periodo de octubre de 2016 a enero de 2017. Se entrevistaron individualmente 221 mujeres climatéricas cardiópatas atendidas en el Ambulatorio de Cardiología. Se realizaron análisis de estadística descriptiva, de asociación con el test no paramétrico de Chi cuadrado de independencia (c2) y test de Correlación de Spearman, con la ayuda del programa SPSS Statistics 20. Resultados: hubo predominio de síntomas climatéricos muy intensos como angustia/ansiedad, 75 (33,9%); agotamiento físico y mental, 61 (27,6%); estado de ánimo depresivo, 54 (24,4%); nerviosismo, 59 (26,7%); y insomnio, 45 (20,4%). Hubo asociación significativa en los síntomas climatéricos psicológicos entre sí y entre los síntomas psicológicos y la depresión. Conclusión: los síntomas climatéricos psicológicos pueden dejar las mujeres más propensas a trastornos emocionales, agraviados por la existencia de una enfermedad crónica como la cardiopatía.

ABSTRACT Objective: to investigate psychological climacteric symptoms in women with heart disease. Method: a quantitative study, carried out at a University Hospital in the Northeast of Brazil, from October 2016 to January 2017. A total of 221 climacteric women with heart disease were interviewed at the Cardiology Outpatient Clinic. Descriptive statistical analysis was performed, association with the non-parametric Chi-square test of independence (c2) and Spearman's correlation test, with the use of the SPSS Statistics 20 program. Results: there was a predominance of very intense climacteric symptoms such as anguish/anxiety, 75 (33.9%); physical and mental exhaustion, 61 (27.6%); depressed mood, 54 (24.4%); nervousness, 59 (26.7%); and insomnia, 45 (20.4%). There were significant associations among the psychological climacteric symptoms and between the psychological symptoms and depression. Conclusion: psychological climacteric symptoms appear to make women more prone to emotional disorders, aggravated by the existence of a chronic disease such as cardiopathy.