Central and Systemic Responses to Methionine-Induced Hyperhomocysteinemia in Mice

Hyperhomocysteinemia has been considered a risk factor for neuropsychiatric disorders, but the mechanisms involved in this process have not been completely elucidated. the aim of this study was to analyze the influence of hyperhomocysteinemia induction by methionine supplementation considering different levels and periods of exposure in mice. for this purpose, methionine supplementation at concentrations of 0.5 and 1% were administered in water to increase homocysteinemia in male C57BL/6 mice, and was maintained for 3 time periods (2, 4 and 6 months of treatment). the results from one-carbon metabolism parameters, brain-derived neurotrophic factor (BDNF) concentrations and behavioral evaluation were compared. the 0.5% supplementation was efficient in increasing plasma homocysteine levels after 2 and 6 months. the 1% supplementation, increased plasma homocysteine after 2, 4 and 6 months. Little influence was observed in cysteine and glutathione concentrations. Frontal cortex BDNF levels showed a lack of treatment influence in all periods; only the expected decrease due to increasing age was observed. Moreover, the only behavioral alteration observed using a novel object recognition task was that which was expected with increasing age. We found that responses to hyperhomocysteinemia varied based on how it was reached, and the length of toxicity. Moreover, hyperhomocysteinemia can affect the normal pattern of one carbon metabolism during age increase in mice. These findings allow the establishment of a reliable animal model for studies in this field.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilFAPESP: 2010/00075-2FAPESP: 2011/15699-4Web of Scienc

Serum homocysteine and cysteine levels and associated factors in children and adolescents

Introduction: Studies have identified high serum homocysteine (Hcy) and cysteine (Cys) levels as risk factor for cardiovascular diseases, important cause of death in all world. The factors associated with high levels of these biochemistry markers in adults are well known; however, data are sparse on these associations in the pediatric age group. Objective: the objective was to identify factors associated with different concentrations of serum Hcy and Cys in children and adolescents. Methods: a cross-sectional study with 483 individuals of 7-15 years of age of both sexes, from a municipality of Bahia. Serum Hcy and Cys levels were considered outcome variables, with exposure being evaluated according to sociodemographic, clinical, biochemical and lifestyle variables. Polytomous logistic regression was used to evaluate the association between exposure and outcome. Results: high serum Hcy levels were associated with being male (PR=3.74; p12 years, being overweight, high blood pressure, low HDL-c levels, high triglyceride levels and poor intake of protective foods are common factors found in individuals with increased serum Hcy and Cys levels. Being male was associated with high serum Hcy levels alone. Considering that these factors are already present early in life, measures should be adopted to prevent and control high Hcy and Cys levels, promoting health and preventing chronic non-communicable diseases at this stage of life

Sleepiness, inflammation and oxidative stress markers in middle-aged males with obstructive sleep apnea without metabolic syndrome: a cross-sectional study

Background: the simultaneous occurrence of metabolic syndrome and excessive daytime sleepiness are very common in obstructive sleep apnea (OSA) patients. Both conditions, if present in OSA, have been reported to be associated with inflammation and disruption of oxidative stress balance that impair the cardiovascular system. To verify the impact of daytime sleepiness on inflammatory and oxidative stress markers, we evaluated OSA patients without significant metabolic disturbance.Methods: Thirty-five male subjects without diagnostic criteria for metabolic syndrome (Adult Treatment Panel III) were distributed into a control group (n = 10) (43 +/- 10.56 years, apnea-hypopnea index - AHI 2.71 +/- 1.48/hour), a non-sleepy OSA group (n = 11) (42.36 +/- 9.48 years, AHI 29.48 +/- 22.83/hour) and a sleepy OSA group (n = 14) (45.43 +/- 10.06 years, AHI 38.20 +/- 25.54/hour). Excessive daytime sleepiness was considered when Epworth sleepiness scale score was >= 10. Levels of high-sensitivity C-reactive protein, homocysteine and cysteine, and paraoxonase-1 activity and arylesterase activity of paraoxonase-1 were evaluated.Results: Patients with OSA and excessive daytime sleepiness presented increased high-sensitivity C-reactive protein levels even after controlling for confounders. No significant differences were found among the groups in paraoxonase-1 activity nor arylesterase activity of paraoxonase-1. AHI was independently associated and excessive daytime sleepiness tended to have an association with high-sensitivity C-reactive protein.Conclusions: in the absence of metabolic syndrome, increased inflammatory response was associated with AHI and daytime sleepiness, while OSA was not associated with abnormalities in oxidative stress markers.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Associacao Fundo de Incentivo a Pesquisa (AFIP)Universidade Federal de São Paulo UNIFESP EPM, Dept Psychobiol, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP EPM, Dept Med, São Paulo, BrazilFac Med ABC FUABC, Dept Morphol & Physiol, Santo Andre, SP, BrazilUniversidade Federal de São Paulo UNIFESP EPM, Dept Psychobiol, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP EPM, Dept Med, São Paulo, BrazilFAPESP: 98/14303-3CNPq: 501343/2010-5Web of Scienc

Analysis of male reproductive parameters in a murine model of mucopolysaccharidosis type I (MPS I)

Mucopolysaccharidosis (MPS) I is a lysosomal storage disorder (LSD) that is characterised by alpha-L-idu-ronidase (Idua) deficiency and continuous deposition of glycosaminoglycans (GAGs), which consequently interferes with cell signalling mechanisms and results in multisystemic and progressive symptoms. The animal model of MPS I (Idua-/-) has been widely studied to elucidate the consequences and progression of the disorder; however, studies specifically assessing the male reproductive tract are lacking. The aim of this study was to evaluate some of the reproductive characteristics of male MPS I mice in two phases of life. Reproductive organ biometry, sperm counts, sperm morphological evaluation, plasma testosterone measurements and histopathological, histomorphometrical and immunohistochemical analysis were performed in 3- and 6-month-old C57BL/6 Idua+/+ and Idua-/- mice. Seminal vesicle weights were decreased in both the 3- and 6-month-old Idua-/- mice. Decrease in sperm counts and the majority of the histopathological signs were observed in the 6-month-old Idua-/- mice. No differences were detected in the sperm morphological analysis. Immunohistochemistry revealed that seminiferous tubules from 3-month-old Idua-/- mice were more intensely stained with anti-caspase-3 than 3-month-old Idua+/+ mice, but no difference was found at 6 months. These results suggest that MPS I interferes with male reproductive parameters both in 3 and 6-month-old animals and histopathological signs are more pronounced in 6-month-old mice, indicating that the effects of the disorder may intensify with the disease progression.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)AFIPFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed Sao Paulo, Dept Psychobiol, BR-04024002 Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Biosci, Santos, SP, BrazilUniv Fed Sao Paulo, Dept Psychobiol, BR-04024002 Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Biosci, Santos, SP, BrazilWeb of Scienc

Subchronical treatment with Fluoxetine modifies the activity of the MCHergic and hypocretinergic systems. Evidences from peptide CSF concentration and gene expression

In the postero-lateral hypothalamus are located two neuronal systems that utilize the neuropeptides melanin-concentrating hormone (MCH) and hypocretins (also called orexins) as neuromodulators. These systems have reciprocal connections between them, and project throughout the central nervous system. MCH has been involved in the generation of sleep, mainly REM sleep, while hypocretins have a critical role in the generation of wakefulness. MCHergic activity is also involved in the pathophysiology of major depressive disorder (MD). In this regards, intracerebral administration of MCH promotes pro-depressive behaviors (i.e., immobility in the forced swimming test) and REM sleep hypersomnia, which is an important trait of depression. Furthermore, the antagonism of the MCHR-1 receptor has a reliable antidepressant effect, suggesting that MCH is a pro-depressive factor. Hypocretins have been also involved in mood regulation; however, their role in depression is still on debate. Taking these data into account, we explored whether systemic subchronical treatment with Fluoxetine (FLX), a serotonergic antidepressant, modifies the concentration of MCH in the cerebrospinal fluid (CSF), as well as the preproMCH mRNA expression. We also evaluated the hypocretinergic system by quantifying the hypocretin-levels in the CSF and the preprohypocretin mRNA expression. Compared to control, FLX increased the levels of preprohypocretin mRNA without affecting the hypocretin-1 CSF levels. On the contrary, FLX significantly decreased the MCH CSF concentration without affecting the preproMCH gene expression. This result is in agreement with the fact that MCH serum level diminishes during the antidepressant treatment in MD, and supports the hypothesis that an increase in the MCHergic activity could have pro-depressive consequences. (C) 2016 Brazilian Association of Sleep. Production and Hosting by Elsevier B.V.Proyecto de Cooperacion Bilateral Uruguay- Brasil, Dicyt-CNPqPrograma de Desarrollo de Ciencias Basicas (PEDECIBA)Associacao Fundo de Incentivo a Pesquisa (AFIP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Univ Fed Sao Paulo, Dept Psychobiol, Sao Paulo, BrazilUniv Republica, Dept Physiol, Sch Med, Montevideo, UruguayUniv Fed Sao Paulo, Dept Psychobiol, Sao Paulo, BrazilProyecto de Cooperacion Bilateral Uruguay- Brasil, Dicyt-CNPq: ANII-FCE-1-2011-1-5997Web of Scienc

Sleep deprivation impairs calcium signaling in mouse splenocytes and leads to a decreased immune response

Background: Sleep is a physiological event that directly influences health by affecting the immune system, in which calcium (Ca2+) plays a critical signaling role. We performed live cell measurements of cytosolic Ca2+ mobilization to understand the changes in Ca2+ signaling that occur in splenic immune cells after various periods of sleep deprivation (SD).Methods: Adult male mice were subjected to sleep deprivation by platform technique for different periods (from 12 to 72 h) and Ca2+ intracellular fluctuations were evaluated in splenocytes by confocal microscopy. We also performed spleen cell evaluation by flow cytometry and analyzed intracellular Ca2+ mobilization in endoplasmic reticulum and mitochondria. Additionally. Ca2+ channel gene expression was evaluatedResults: Splenocytes showed a progressive loss of intracellular Ca2+ maintenance from endoplasmic reticulum (ER) stores. Transient Ca2+ buffering by the mitochondria was further compromised. These findings were confirmed by changes in mitochondrial integrity and in the performance of the store operated calcium entry (SOCE) and stromal interaction molecule 1 (STIM1) Ca2+ channels.Conclusions and general significance: These novel data suggest that SD impairs Ca2+ signaling, most likely as a result of ER stress, leading to an insufficient Ca2+ supply for signaling events. Our results support the previously described immunosuppressive effects of sleep loss and provide additional information on the cellular and molecular mechanisms involved in sleep function. (C) 2012 Elsevier B.V. All rights reserved.AFIP (Associacao Fundo de Incentivo a Pesquisa)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo UNIFESP, Dept Psychobiol, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Biosci, Santos, SP, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Biochem, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Psychobiol, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Biosci, Santos, SP, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Biochem, São Paulo, BrazilFAPESP: 05/04366-3FAPESP: CEPID 98/14303-6Web of Scienc

Tryptophan overloading activates brain regions involved with cognition, mood and anxiety

Tryptophan is the only precursor of serotonin and mediates serotonergic activity in the brain. Previous studies have shown that the administration of tryptophan or tryptophan depletion significantly alters cognition, mood and anxiety. Nevertheless, the neurobiological alterations that follow these changes have not yet been fully investigated. The aim of this study was to verify the effects of a tryptophan-enriched diet on immunoreactivity to Fos-protein in the rat brain. Sixteen male Wistar rats were distributed into two groups that either received standard chow diet or a tryptophan-enriched diet for a period of thirty days. On the morning of the 31st day, animals were euthanized and subsequently analyzed for Fos-immunoreactivity (Fos-ir) in the dorsal and median raphe nuclei and in regions that receive serotonin innervation from these two brain areas. Treatment with a tryptophan-enriched diet increased Fos-ir in the prefrontal cortex, nucleus accumbens, paraventricular hypothalamus, arcuate and ventromedial hypothalamus, dorsolateral and dorsomedial periaqueductal grey and dorsal and median raphe nucleus. These observations suggest that the physiological and behavioral alterations that follow the administration of tryptophan are associated with the activation of brain regions that regulate cognition and mood/anxiety-related responses.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ Fed Sao Paulo, Dept Psicobiol, Rua Napoleao Barros 925,3 Andar, BR-04023062 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Dept Biociencias, Rua Silva Jardim 136,3 Andar, BR-11060001 Santos, SP, BrazilUniv Fed Sao Paulo, Dept Psicobiol, Rua Napoleao Barros 925,3 Andar, BR-04023062 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Dept Biociencias, Rua Silva Jardim 136,3 Andar, BR-11060001 Santos, SP, BrazilWeb of Scienc

Effect of Vitamin B Deprivation during Pregnancy and Lactation on Homocysteine Metabolism and Related Metabolites in Brain and Plasma of Mice Offspring

Epidemiological and experimental studies indicate that the altered fetal and neonatal environment influences physiological functions and may increase the risk of developing chronic diseases in adulthood. Because homocysteine (Hcy) metabolic imbalance is considered a risk factor for neurodegenerative diseases, we investigated whether maternal Vitamin B deficiency during early development alters the offspring's methionine-homocysteine metabolism in their brain. To this end, the dams were submitted to experimental diet one month before and during pregnancy or pregnancy/lactation. After birth, the offspring were organized into the following groups: control (CT), deficient diet during pregnancy and lactation (DPL) and deficient diet during pregnancy (DP). the mice were euthanized at various stages of development. Hcy, cysteine, glutathione (GSH), S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), folate and cobalamin concentrations were measured in the plasma and/or brain. At postnatal day (PND) 0, total brain of female and male offspring exhibited decreased SAM/SAH ratios. Moreover, at PND 28, we observed decreased GSH/GSSG ratios in both females and males in the DPL group. Exposure to a Vitamin B-deficient diet during the ontogenic plasticity period had a negative impact on plasma folate and brain cortex SAM concentrations in aged DPL males. We also observed decreased plasma GSH concentrations in both DP and DPL males (PND 210). Additionally, this manipulation seemed to affect the female and male offspring differently. the decreased plasma GSH concentration may reflect redox changes in tissues and the decreased brain cortex SAM may be involved in changes of gene expression, which could contribute to neurodegenerative diseases over the long term.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)AFIPUniversidade Federal de São Paulo, Dept Pediat, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilUniv São Paulo, Sch Pharmaceut Sci, São Paulo, BrazilUniv Pernambuco, Pediat Hematol & Oncol Ctr, Inst Biol Sci, Recife, PE, BrazilUniversidade Federal de São Paulo, Dept Pediat, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilFAPESP: 2010/00075-2Web of Scienc