Brain Mitochondria as a Therapeutic Target for Carnosic Acid.

Carnosic acid (CA), a diterpene obtained mainly from Rosmarinus officinalis and Salvia officinalis, exerts antioxidant, anti-inflammatory, and anti-apoptotic effects in mammalian cells. At least in part, those benefits are associated with the ability that CA modulates mitochondrial physiology. CA attenuated bioenergetics collapse and redox impairments in the mitochondria obtained from brain cells exposed to several toxicants in both in vitro and in vivo experimental models. CA is a potent inducer of the major modulator of the redox biology in animal cells, the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), which controls the expression of a myriad of genes whose products are involved with cytoprotection in different contexts. Moreover, CA upregulates signaling pathways related to the degradation of damaged mitochondria (mitophagy) and with the synthesis of these organelles (mitochondrial biogenesis). Thus, CA may be considered an agent that induces mitochondrial renewal, depending on the circumstances. In this review, we discuss about the mechanisms of action by which CA promotes mitochondrial protection in brain cells.

Effects of Extracts of Two Selected Strains of Haematococcus pluvialis on Adipocyte Function.

Recently, microalgae are arousing considerable interest as a source of countless molecules with potential impacts in the nutraceutical and pharmaceutical fields. Haematococcus pluvialis, also named Haematococcus lacustris, is the largest producer of astaxanthin, a carotenoid exhibiting powerful health effects, including anti-lipogenic and anti-diabetic activities. This study was carried out to investigate the properties of two selected strains of H. pluvialis (FBR1 and FBR2) on lipid metabolism, lipolysis and adipogenesis using an in vitro obesity model. FBR1 and FBR2 showed no antiproliferative effect at the lowest concentration in 3T3-L1 adipocytes. Treatment with FBR2 extract reduced lipid deposition, detected via Oil Red O staining and the immunocontent of the adipogenic proteins PPARγ, ACLY and AMPK was revealed using Western blot analysis. Extracts from both strains induced lipolysis in vitro and reduced the secretion of interleukin-6 and tumor necrosis factor-α. Moreover, the FBR1 and FBR2 extracts improved mitochondrial function, reducing the levels of mitochondrial superoxide anion radical and increasing mitochondrial mass compared to untreated adipocytes. These findings suggest that FBR2 extract, more so than FBR1, may represent a promising strategy in overweight and obesity prevention and treatment.

Testing the Accuracy of Wearable Technology to Assess Sleep Behaviour in Domestic Dogs: A Prospective Tool for Animal Welfare Assessment in Kennels.

Sleep is a physiological process that has been shown to impact both physical and psychological heath of individuals when compromised; hence, it has the potential to be used as an indicator of animal welfare. Nonetheless, evaluating sleep in non-human species normally involves manipulation of the subjects (i.e., placement of electrodes on the cranium), and most studies are conducted in a laboratory setting, which limits the generalisability of information obtained, and the species investigated. In this study, we evaluated an alternative method of assessing sleep behaviour in domestic dogs, using a wearable sensor, and compared the measurements obtained to behavioural observations to evaluate accuracy. Differences between methods ranged from 0.13% to 59.3% for diurnal observations and 0.1% to 95.9% for nocturnal observations for point-by-point observations. Comparisons between methods showed significant differences in certain behaviours, such as inactivity and activity for diurnal recordings. However, total activity and total sleep recorded did not differ statistically between methods. Overall, the wearable technology tested was found to be a useful, and a less-time consuming, tool in comparison to direct behavioural observations for the evaluation of behaviours and their indication of wellbeing in dogs. The agreement between the wearable technology and directly observed data ranged from 75% to 99% for recorded behaviours, and these results are similar to previous findings in the literature.

Impact of Maternal Folic Acid Supplementation on Descendants' Kidney in Adulthood.

Supplementation with folic acid (FA) during gestation has been recommended by medical society all over the world, but some studies have shown that intake of high folic acid diet may unleash damages to the descendants. OBJECTIVES: Describing the effects of maternal supplementation with FA during gestation on offspring's kidney at late life stages. DATA SOURCE: It is a systematic review by which were consulted the following databases: Medline, through Pubmed, Lilacs, and SciELO. The research was performed using the keywords "Folic acid", "Gestation" and "Kidney". STUDY SELECTION: Eight studies were regarded for this systematic review. DATA COLLECTION: Only studies that evaluated folic acid consumption during gestation and its effects exclusively on descendants' kidney at several phases of life were regarded. RESULTS: Gestational FA intake did not change the renal volume, glomerular filtration rate and the expression of some essential genes in the kidney of puppies whose dams were supplemented with FA. Maternal consumption of double FA plus selenium diet was effective in preserving antioxidant enzymes activity in the kidney of descendants from mothers exposed to alcohol. FA supplementation decreased some gross anomalies in the puppies caused by teratogenic drug despite of had not been effective in preventing some renal architectural damages. CONCLUSION: FA supplementation did not cause renal toxicity; it exerted an antioxidant protective effect and mitigated some renal disorders caused by severe aggressions.

A suplementação com ácido fólico (AF) durante a gestação tem sido recomendada pela sociedade médica em todo o mundo, mas alguns estudos têm mostrado que a ingestão de altas quantidades de ácido fólico na dieta pode desencadear danos aos descendentes. Objetivos: Descrever os efeitos da suplementação materna com AF durante a gestação no rim da prole em fases tardias da vida. Fonte de Dados: Trata-se de uma revisão sistemática realizada através da consulta das seguintes bases de dados: Medline, através da Plataforma Pubmed, Lilacs e Scielo. A pesquisa foi realizada utilizando-se as palavras-chave "Ácido Fólico", "Gestação" e "Rim". Seleção dos Estudos: Oito estudos foram considerados para esta revisão sistemática. Coleta de Dados: Foram incluídos estudos que abordaram o consumo de ácido fólico durante a gestação e seus efeitos exclusivamente no rim dos descendentes em diferentes fases da vida. Resultados: O consumo gestacional de AF não alterou o volume renal, a taxa de filtração glomerular e a expressão de alguns genes essenciais no rim dos filhotes de mães suplementadas com AF. A associação de AF e selênio na dieta materna foi eficaz na preservação da atividade de enzimas antioxidantes no rim da prole de mães expostas ao álcool. O consumo de AF diminuiu algumas anomalias importantes nos filhotes causadas por drogas teratogênicas, apesar de não ter sido eficiente na prevenção de alguns danos a arquitetura renal. Conclusão: A suplementação com AF não causou toxicicdade renal, exerceu efeito protetor antioxidante e mitigou algumas desordens renais causadas por agressões severas.

The isothiocyanate sulforaphane prevents mitochondrial impairment and neuroinflammation in the human dopaminergic SH-SY5Y and in the mouse microglial BV2 cells: role for heme oxygenase-1.

Sulforaphane (SFN) promotes protective effects in different cell types. Nonetheless, it remains to be clarified by which mechanism SFN exerts benefits in mammalian cells. Mitochondria are a major source of adenosine triphosphate (ATP) and reactive species in nucleated cells. Mitochondrial impairment result in cellular redox biology disruption, bioenergetic status collapse, and inflammation. Evidence suggest that mitochondrial dysfunction plays a role in neurological disorders. Since a cure was not discovered yet to some of these diseases, investigating strategies to promote mitochondrial protection is pharmacologically relevant and may improve life quality of patients suffering from these maladies. Natural molecules, such as SFN, are potent inducers of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) and, consequently, stimulate the expression of genes whose products, such as heme oxygenase-1 (HO-1), induce cytoprotective actions in mammalian tissues. In this work, we investigated whether SFN (5 µM) would be capable to prevent the dysfunctions caused by chlorpyrifos (CPF) on the human dopaminergic SH-SY5Y cells. Moreover, we examined the effects of a pretreatment with SFN at the same concentration on the mouse microglial BV2 cells stimulated by lipopolysaccharide (LPS) in an experimental model of neuroinflammation. SFN prevented the mitochondrial impairment and the neuroinflammation caused by the chemical stressors in both cell types. Inhibition of heme oxygenase-1 (HO-1) suppressed the mitochondrial protection and anti-inflammatory action afforded by SFN in this experimental model. Overall, SFN promoted cytoprotection by a mechanism dependent on the HO-1 enzyme in the SH-SY5Y and BV2 cells.

The C-glucosyl flavone isoorientin pretreatment attenuates the methylglyoxal-induced mitochondrial dysfunction in the human neuroblastoma SH-SY5Y cells: role for the AMPK-PI3K/Akt/Nrf2/γ-GCL/GSH axis.

The reactive dicarbonyl methylglyoxal (MG) behaves as a pro-oxidant agent, causing redox dysfunction and cell death by different mechanisms in mammalian cells. MG is also a mitochondrial toxicant, impairing the oxidative phosphorylation (OXPHOS) system and leading to bioenergetics and redox collapses. MG induces glycation and exerts an important role in neurodegenerative and cardiovascular diseases. Isoorientin (ISO), a C-glucosyl flavone found in Aspalathus linearis, Fagopyrum esculentum, and Passiflora edulis, among others, is an antioxidant and anti-inflammatory molecule. ISO is a potent inducer of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), the master modulator of the redox environment in mammals. We investigated here whether ISO would prevent the mitochondria-related redox and bioenergetics impairments induced by MG in the human neuroblastoma SH-SY5Y cells. The cells were administrated with ISO at 20 µM for 18 h prior to the exposure to MG at 500 µM for further 24 h. It was observed that ISO efficiently prevented the mitochondrial impairments caused by MG. ISO upregulated the activity of the enzyme γ-glutamate-cysteine ligase (γ-GCL), consequently stimulating the synthesis of glutathione (GSH). The inhibition of γ-GCL, adenosine monophosphate-activated protein kinase (AMPK), and phosphoinositide 3-kinase/Akt (PI3K/Akt) suppressed the beneficial effects induced by ISO on the MG-challenged cells. Moreover, silencing of Nrf2 blocked the ISO-dependent γ-GCL and GSH upregulation and the effects on the mitochondria of the MG-challenged cells. Then, ISO caused mitochondrial protection by an AMPK-PI3K/Akt/Nrf2/γ-GCL/GSH-dependent manner in MG-administrated SH-SY5Y cells.

Impact of Maternal Folic Acid Supplementation on Descendants' Kidney in Adulthood / Impacto da suplementação materna com ácido fólico no rim dos descendentes na vida adulta

Abstract Supplementation with folic acid (FA) during gestation has been recommended by medical society all over the world, but some studies have shown that intake of high folic acid diet may unleash damages to the descendants. Objectives: Describing the effects of maternal supplementation with FA during gestation on offspring's kidney at late life stages. Data Source: It is a systematic review by which were consulted the following databases: Medline, through Pubmed, Lilacs, and SciELO. The research was performed using the keywords "Folic acid", "Gestation" and "Kidney". Study Selection: Eight studies were regarded for this systematic review. Data Collection: Only studies that evaluated folic acid consumption during gestation and its effects exclusively on descendants' kidney at several phases of life were regarded. Results: Gestational FA intake did not change the renal volume, glomerular filtration rate and the expression of some essential genes in the kidney of puppies whose dams were supplemented with FA. Maternal consumption of double FA plus selenium diet was effective in preserving antioxidant enzymes activity in the kidney of descendants from mothers exposed to alcohol. FA supplementation decreased some gross anomalies in the puppies caused by teratogenic drug despite of had not been effective in preventing some renal architectural damages. Conclusion: FA supplementation did not cause renal toxicity; it exerted an antioxidant protective effect and mitigated some renal disorders caused by severe aggressions.

Resumo A suplementação com ácido fólico (AF) durante a gestação tem sido recomendada pela sociedade médica em todo o mundo, mas alguns estudos têm mostrado que a ingestão de altas quantidades de ácido fólico na dieta pode desencadear danos aos descendentes. Objetivos: Descrever os efeitos da suplementação materna com AF durante a gestação no rim da prole em fases tardias da vida. Fonte de Dados: Trata-se de uma revisão sistemática realizada através da consulta das seguintes bases de dados: Medline, através da Plataforma Pubmed, Lilacs e Scielo. A pesquisa foi realizada utilizando-se as palavras-chave "Ácido Fólico", "Gestação" e "Rim". Seleção dos Estudos: Oito estudos foram considerados para esta revisão sistemática. Coleta de Dados: Foram incluídos estudos que abordaram o consumo de ácido fólico durante a gestação e seus efeitos exclusivamente no rim dos descendentes em diferentes fases da vida. Resultados: O consumo gestacional de AF não alterou o volume renal, a taxa de filtração glomerular e a expressão de alguns genes essenciais no rim dos filhotes de mães suplementadas com AF. A associação de AF e selênio na dieta materna foi eficaz na preservação da atividade de enzimas antioxidantes no rim da prole de mães expostas ao álcool. O consumo de AF diminuiu algumas anomalias importantes nos filhotes causadas por drogas teratogênicas, apesar de não ter sido eficiente na prevenção de alguns danos a arquitetura renal. Conclusão: A suplementação com AF não causou toxicicdade renal, exerceu efeito protetor antioxidante e mitigou algumas desordens renais causadas por agressões severas.

Short-term effectiveness of transcranial direct current stimulation in the nociceptive behavior of neuropathic pain rats in development.

Neuropathic pain (NP) is caused by a lesion that triggers pain chronification and central sensitization and it can develop in a different manner, dependent of age. Recent studies have demonstrated the efficacy of transcranial direct current stimulation (tDCS) for treating NP. Then, we aimed to investigate the effects of tDCS and BDNF levels in neuropathic pain rats in development, with 30 days old in the beginning of experiments. Eight-five male Wistar rats were subjected to chronic constriction injury. After establishment of NP, bimodal tDCS was applied to the rats for eight consecutive days, for 20 minutes each session. Subsequently, nociceptive behavior was assessed at baseline, 14 days after surgery, 1 day and 7 days after the end of tDCS. The rats were sacrificed 8 days after the last session of tDCS. An increase in the nociceptive threshold was observed in rats in development 1 day after the end of tDCS (short-term effect), but this effect was not maintained 7 days after the end of tDCS (long-term effect). Furthermore, brain derived neurotrophic factor (BDNF) levels were analyzed in the frontal cortex, spinal cord and serum using ELISA assays. The neuropathic pain model showed an effect of BDNF in the spinal cord of rats in development. There were no effects of BNDF levels of pain or tDCS in the frontal cortex or serum. In conclusion, tDCS is an effective technique to relieve nociceptive behavior at a short-term effect in neuropathic pain rats in development, and BDNF levels were not altered at long-term effect.

How environmental conditions affect sleep? An investigation in domestic dogs (Canis lupus familiaris).

Studies with humans and some other animal species have shown that sleep is compromised when the presence of external factors such as light, sound, and temperature surpass normal levels. This study investigated the effects of these environmental conditions on 13 kennelled laboratory dogs, assessing whether each variable interfered with their sleep behaviour and/or increased stress responses, which could further compromise sleep quality. The behaviour of dogs was video recorded for eight months. Diurnal and nocturnal behaviour were recorded, along with naturally occurring levels of temperature, light and sound in the dogs' kennel environment. Faecal cortisol metabolites (FCM), from samples collected every morning, were used to monitor the dogs' adrenocortical activity. GLMM models and non-parametric tests were conducted to evaluate the relationship between sleeping patterns, environmental variables, and stress on the studied dogs. Nocturnal sleep decreased in response to increases in temperature and in day light duration. No effects of sound and FCM levels on dogs' sleep were observed. However, diurnal sleep was affected by sound and FCM levels, decreasing when both factors increased. Additionally, noisier days increased stress responses, especially in male dogs. Increased FCM levels were associated with changes in the diurnal behaviour of dogs; for example, decreased activity. The decrease in daily activities and increased physiological stress responses could be associated with maladaptation to the environment, which could indicate poor welfare. Our study suggests that mitigating the impact of environmental conditions in the kennels could improve sleep quality and the overall quality of life of the dogs.

A Pretreatment with Isoorientin Attenuates Redox Disruption, Mitochondrial Impairment, and Inflammation Caused by Chlorpyrifos in a Dopaminergic Cell Line: Involvement of the Nrf2/HO-1 Axis.

The C-glucosyl flavone isoorientin (ISO) is obtained by humans from the diet and exhibits several cytoprotective effects, as demonstrated in different experimental models. However, it was not previously shown whether ISO would be able to prevent mitochondrial impairment in cells exposed to a chemical stressor. Thus, we treated the human neuroblastoma SH-SY5Y cells with ISO (0.5-20 µM) for 18 h before a challenge with chlorpyrifos (CPF) at 100 µM for additional 24 h. We observed that ISO prevented the CPF-induced lipid peroxidation and protein carbonylation and nitration in the membranes of mitochondria extracted from CPF-treated cells. ISO also attenuated the CPF-elicited increase in the production of reactive species in this experimental model. Moreover, ISO prevented the CPF-induced disruption in the activity of components of the oxidative phosphorylation (OXPHOS) system in the SH-SY5Y cells. ISO also promoted an anti-inflammatory action in the cells exposed to CPF. CPF caused a decrease in the activity of the enzyme heme oxygenase-1 (HO-1), a cytoprotective agent. On the other hand, ISO upregulated HO-1 activity in SH-SY5Y cells. Inhibition of HO-1 by zinc protoporphyrin-IX (ZnPP-IX) suppressed the cytoprotection induced by ISO in the CPF-treated cells. Besides, silencing of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) abolished the ISO-induced HO-1 upregulation and mitochondrial benefits induced by this flavone on the CPF-challenged cells. Thus, ISO protected mitochondria of the CPF-treated cells by an Nrf2/HO-1-dependent fashion in the SH-SY5Y cells.

The cyclic interaction between daytime behavior and the sleep behavior of laboratory dogs.

Sleep deprivation has been found to negatively affect an individual´s physical and psychological health. Sleep loss affects activity patterns, increases anxiety-like behaviors, decreases cognitive performance and is associated with depressive states. The activity/rest cycle of dogs has been investigated before, but little is known about the effects of sleep loss on the behavior of the species. Dogs are polyphasic sleepers, meaning the behavior is most observed at night, but bouts are also present during the day. However, sleep can vary with ecological and biological factors, such as age, sex, fitness, and even human presence. In this study, kennelled laboratory adult dogs' sleep and diurnal behavior were recorded during 24-h, five-day assessment periods to investigate sleep quality and its effect on daily behavior. In total, 1560 h of data were analyzed, and sleep metrics and diurnal behavior were quantified. The relationship between sleeping patterns and behavior and the effect of age and sex were evaluated using non-parametric statistical tests and GLMM modelling. Dogs in our study slept substantially less than previously reported and presented a modified sleep architecture with fewer awakenings during the night and almost no sleep during the day. Sleep loss increased inactivity, decreased play and alert behaviors, while increased time spent eating during the day. Males appeared to be more affected by sleep fragmentation than females. Different age groups also experienced different effects of sleep loss. Overall, dogs appear to compensate for the lack of sleep during the night by remaining inactive during the day. With further investigations, the relationship between sleep loss and behavior has the potential to be used as a measure of animal welfare.

Sesamol prevents mitochondrial impairment and pro-inflammatory alterations in the human neuroblastoma SH-SY5Y cells: role for Nrf2.

Mitochondria are a primary source and a target of reactive oxygen species (ROS). Increased mitochondrial production of ROS is associated with bioenergetics decline, cell death, and inflammation. Here we investigated whether a pretreatment (for 24 h) with sesamol (SES; at 12.5-50 µM) would be efficient in preventing the mitochondrial collapse induced by hydrogen peroxide (H2O2, at 300 µM) in the human neuroblastoma SH-SY5Y cell line. We have found that a pretreatment with SES at 25 µM decreased the effects of H2O2 on lipid peroxidation, protein carbonylation, and protein nitration in membranes obtained from the mitochondria isolated from the SH-SY5Y cells. In this regard, SES pretreatment decreased the production of superoxide anion radical (O2-•) by the mitochondria of H2O2-treated cells. SES also prevented the mitochondrial dysfunction induced by H2O2, as assessed by analyzing the activity of the complexes I and V. The H2O2-induced reduction in the production of adenosine triphosphate (ATP) was also prevented by SES. The levels of the pro-inflammatory cytokines interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α), as well as the activity of the transcription factor nuclear factor-κB (NF-κB) were downregulated by the SES pretreatment in the H2O2-challenged cells. Silencing of the nuclear factor erythroid 2-related factor 2 (Nrf2) transcription factor abolished the protection induced by SES regarding mitochondrial function and inflammation. Thus, SES depends on Nrf2 to promote mitochondrial protection in cells facing redox impairment.

Suppression of Mitochondria-Related Bioenergetics Collapse and Redox Impairment by Tanshinone I, a Diterpenoid Found in Salvia miltiorrhiza Bunge (Danshen), in the Human Dopaminergic SH-SY5Y Cell Line Exposed to Chlorpyrifos.

Tanshinone I (T-I, C18H12O3) is a diterpene found in Salvia miltiorrhiza Bunge (Danshen) and promotes cytoprotection in several experimental models. Chlorpyrifos (CPF) is an agrochemical that causes bioenergetics failure, redox impairment, inflammation, and cell death in animal tissues. Here, we investigated whether T-I would be able to prevent the consequences resulting from the exposure of the human dopaminergic SH-SY5Y cells to CPF. We found that a pretreatment with T-I at 2.5 µM for 2 h suppressed lipid peroxidation and protein carbonylation and nitration on the membranes of mitochondria extracted from the CPF-treated cells. Also, T-I reduced the production of radical superoxide (O2-•) by the mitochondria of the CPF-challenged cells. The production of nitric oxide (NO•) and hydrogen peroxide (H2O2) was also decreased by T-I in the cells exposed to CPF. The CPF-induced decrease in the activity of the complexes I-III, II-III, and V was abolished by a pretreatment with T-I. Loss of mitochondrial membrane potential (ΔΨm) and reduction in the production of adenosine triphosphate (ATP) were also prevented by T-I in the CPF-treated cells. T-I also induced anti-inflammatory effects in the CPF-treated cells by decreasing the levels of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) and the activity of the nuclear factor-κB (NF-κB). Inhibition of heme oxygenase-1 (HO-1) or silencing of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) blocked the T-I-promoted mitochondrial protection and anti-inflammatory action. Overall, T-I depended on the Nrf2/HO-1 axis to prevent the deleterious effects caused by CPF in this experimental model.

Pinocembrin pretreatment counteracts the chlorpyrifos-induced HO-1 downregulation, mitochondrial dysfunction, and inflammation in the SH-SY5Y cells.

Chlorpyrifos (CPF), an insecticide, induces pro-oxidant, pro-inflammatory, and pro-apoptotic effects in animal cells. Contamination with CPF occurs not only in farms, since CPF is found in the food consumed in homes. Recently, it was demonstrated that CPF affects the mitochondria, inhibiting components of the electron transfer chain (ETC), causing loss of mitochondrial membrane potential (MMP), and reducing the synthesis of adenosine triphosphate (ATP) by the Complex V. Pinocembrin (PB) is found in propolis and exhibits antioxidant, anti-inflammatory, and anti-apoptotic effects in mammalian cells. PB is a potent inducer of the nuclear factor erythroid 2-related factor 2 (Nrf2), which is a major transcription factor controlling the expression of heme oxygease-1 (HO-1), among others. In the present work, we investigated whether PB would be able to prevent the mitochondrial and immune dysfunctions in the human neuroblastoma SH-SY5Y cells exposed to CPF. PB was tested at 1-25 µM for 4 h before the administration of CPF at 100 µM for additional 24 h. We found that PB prevented the CPF-induced inhibition of ETC, loss of MMP, and decline in the ATP synthesis. PB also promoted anti-inflammatory actions in this experimental model. Silencing of Nrf2 or inhibition of HO-1 suppressed the PB-induced effects in the CPF-challenged cells. Thus, PB promoted beneficial effects by a mechanism dependent on the Nrf2/HO-1/CO + BR axis in the CPF-treated cells.

Astaxanthin prevents mitochondrial impairment in the dopaminergic SH-SY5Y cell line exposed to glutamate-mediated excitotoxicity: Role for the Nrf2/HO-1/CO-BR axis.

Mitochondrial dysfunction has been viewed in several diseases, including neurological disorders. In the glutamate (GLU)-mediated excitotoxicity, it has been described mitochondrial impairment, disrupted redox environment, and increased rates of cell death in the affected brain areas. Astaxanthin (AST) is a potent antioxidant and anti-inflammatory xanthophyll that also promotes beneficial mitochondria-related effects in brain cells. However, it is not completely clear how AST would be able to promote mitochondrial protection in those cell types. Thus, we investigated here how AST would protect mitochondria in the dopaminergic SH-SY5Y cell line exposed to GLU. AST was administrated to the cells at 1-40 µM for 24 h prior to the exposure to GLU at 80 mM for additional 24 h. AST prevented the GLU-induced impairment in the activity of the Complexes I and V, the loss in mitochondrial membrane potential (MMP), and the decline in the synthesis of ATP. AST also induced an antioxidant effect in the membranes of mitochondria obtained from the GLU-treated SH-SY5Y cells. Inhibition of the enzyme heme oxygenase-1 (HO-1) or silencing of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) suppressed the AST-promoted cellular and mitochondrial protection. Either tricarbonyldichlororuthenium(II) dimer (CORM-2, a source of carbon monoxide - CO) or bilirubin (BR), that are products of the HO-1-biliverdin reductase (BVR) axis, blocked some of the effects caused by GLU in the SH-SY5Y cells. Overall, our data demonstrate that AST prevented mitochondrial dysfunction by a mechanism related to the Nrf2/HO-1 axis in GLU-challenged cells.

Envolvimento acadêmico, funcionamento adaptativo, problemas internalizantes e problemas externalizantes em universitários / Academic involvement, adaptive functioning, internalizing problems and externalizing problems in college students

Introdução: coincidente com distanciamento da família e interação com novos grupos sociais, a etapa da universidade traz oportunidades para incremento da autonomia frente às tarefas da vida adulta. A literatura aponta dificuldades psicológicas ocorrendo nos primeiros anos universitários, que podem se refletir negativamente no desempenho acadêmico. Objetivo: verificar condições psicológicas de universitários do primeiro ano com respeito a funcionamento adaptativo, problemas internalizantes, problemas externalizantes e envolvimento acadêmico. Método: participaram da pesquisa 50 alunos de primeiro ano de um curso de bacharelado interdisciplinar, de uma universidade pública. Foram utilizados o Adult Self-Report e a Escala de Envolvimento Acadêmico. A aplicação dos instrumentos ocorreu coletivamente, em salas de aula, nos meses de maio e setembro de 2018. Os dados foram analisados estatisticamente e tratados de forma descritiva e inferencial, pela variável sexo. Resultados: médias dos escores de problemas internalizantes foram indicadas em faixas limítrofe/clínicas, nos homens e nas mulheres avaliados. Encontrada correlação positiva moderada entre envolvimento em atividades obrigatórias e funcionamento adaptativo; e correlação negativa moderada entre problemas externalizantes e envolvimento em atividades obrigatórias. Discussão: problemas internalizantes nos homens investigados (em nível semelhante aos das estudantes) não são comumente reportados na literatura concernente,carecendo demais investigações.(AU)

Introduction: the years at university, wich often coincid with distancing from the family, end up becoming interaction with new social groups, college stage brings opportunities to increase autonomy in relation to the tasks of adult life. Literature points out psychological difficulties that occur in the first university years, which may reflect negatively on academic performance. Objective: to verify psychological characteristics of first year undergraduate college students, with respect to adaptive functioning, internalizing problems, externalizing problems and academic involvement. Method: 50 first year students participated in the study from an interdisciplinary bachelor degree course from a public university. Adult Self-Report and Academic Involvement Scale were used. The instruments were applied in classrooms, collectively, in the months of May and September of 2018. Data was statisticaly treated in a descriptive and inferential manner, by gender variable. Results: mean scores of internalizing problems were indicated in borderline/clinical ranges among men and women in the sample. Moderate positive correlation was found between involvement in mandatory activities and adaptive functioning; and negative moderate correlation between externalizing problems and involvement in mandatory activities. Discussion: internalizing problems in the investigated men (similar to those of the female students) are not commonly reported in the literature, requiring further investigation.(AU)

The signaling pathway PI3K/Akt/Nrf2/HO-1 plays a role in the mitochondrial protection promoted by astaxanthin in the SH-SY5Y cells exposed to hydrogen peroxide.

The mitochondria are the major source of reactive species in the mammalian cells. Hydrogen peroxide (H2O2) is a potent inducer of redox impairment by a mechanism, at least in part, dependent on its ability to impair mitochondrial function. H2O2 plays an important role in several pathological conditions, including neurodegeneration and cardiovascular diseases. Astaxanthin (AST) is a xanthophyll that may be found in microalgae, crustaceans, and salmon and exhibits antioxidant and anti-inflammatory effects in different cell types. Even though there is evidence pointing to a role for AST as mitochondrial protectant agent, it was not clearly demonstrated how this xanthophyll attenuates mitochondrial stress. Therefore, we investigated here whether and how AST would be able to prevent the H2O2-induced mitochondrial dysfunction in the human neuroblastoma SH-SY5Y cells. We found that AST (20 µM) prevented the H2O2-induced loss of mitochondrial membrane potential (MMP) and decrease in the activity of the Complexes I and V. AST pretreatment blocked the mitochondria-related pro-apoptotic effects elicited by H2O2. AST upregulated the enzyme heme oxygenase-1 (HO-1) and the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) by a mechanism dependent on the phosphoinositide 3-kinase/Akt (PI3K/Akt) signaling pathway. Inhibition of the PI3K/Akt or of the HO-1 enzyme abolished the AST-induced mitochondrial protection in cells challenged with H2O2. Silencing of Nrf2 caused similar effects. Thus, we suggest that AST promotes mitochondrial protection by a mechanism dependent on the PI3K/Akt/Nrf2/HO-1 signaling pathway in SH-SY5Y cells exposed to H2O2.

Carnosic acid depends on glutathione to promote mitochondrial protection in methylglyoxal-exposed SH-SY5Y cells.

Methylglyoxal (MG) is an endogenously produced toxicant that induces mitochondrial dysfunction leading to impaired redox biology homeostasis, bioenergetics collapse, and cell death in mammalian cells. However, MG toxicity is particularly relevant to neurons and glia given their chemical and metabolic characteristics. Here, we have investigated whether a pretreatment with carnosic acid (CA) would be able to promote mitochondrial protection in human neuroblastoma SH-SY5Y cells exposed to MG. We found that a pretreatment with CA at 1 µM for 12 h prevented the MG-induced lipid peroxidation and protein carbonylation and nitration in the membranes of mitochondria obtained from the SH-SY5Y cells. CA also prevented the MG-elicited Complexes I and V dysfunction, adenosine triphosphate (ATP) levels decline, and loss of mitochondrial membrane potential (MMP). Moreover, CA also reduced the mitochondrial production of the radical anion superoxide (O2-•) in the MG-challenged cells. We found that CA upregulated the synthesis of glutathione (GSH) by increasing the activity of the γ-glutamylcysteine ligase (γ-GCL). Inhibition of the GSH synthesis by buthionine sulfoximine (BSO) abolished the CA-induced mitochondrial protection. Besides, inhibition of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway, as well as silencing of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), suppressed the CA-stimulated protection and the synthesis of GSH. Thus, CA promoted mitochondrial protection by a PI3K/Akt/Nrf2/γ-GCL/GSH axis in MG-treated SH-SY5Y cells.

The Isothiocyanate Sulforaphane Depends on the Nrf2/γ-GCL/GSH Axis to Prevent Mitochondrial Dysfunction in Cells Exposed to Methylglyoxal.

Methylglyoxal (MG) is a reactive dicarbonyl presenting both endogenous (e.g. glycolysis) and exogenous (e.g. food cooking) sources. MG induces neurotoxicity, at least in part, by affecting mitochondrial function, including a decline in the oxidative phosphorylation (OXPHOS) system activity, bioenergetics failure, and redox disturbances. Sulforaphane (SFN) is an isothiocyanate found mainly in cruciferous vegetables and exerts antioxidant and anti-inflammatory effects in mammalian cells. SFN also decreases mitochondrial vulnerability to several chemical stressors. SFN is a potent activator of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), which is a master regulator of the mammalian redox biology. Here, we have investigated whether and how SFN would be able to prevent the MG-induced mitochondrial collapse in the human neuroblastoma SH-SY5Y cells. The cells were exposed to SFN at 5 µM for 24 h prior to the administration of MG at 500 µM for additional 24 h. We found that SFN prevented the MG-induced OXPHOS dysfunction and mitochondrial redox impairment. SFN stimulated the activity of the enzyme γ-glutamylcysteine ligase (γ-GCL), leading to increased synthesis of glutathione (GSH). Inhibition of γ-GCL with buthionine sulfoximine (BSO) or silencing of Nrf2 using small interfering RNA (siRNA) against this transcription factor reduced the levels of GSH and abolished the mitochondrial protection promoted by SFN in the MG-treated cells. Thus, SFN protected mitochondria of the MG-challenged cells by a mechanism involving the Nrf2/γ-GCL/GSH axis.

Mitochondrial Protection and Anti-inflammatory Effects Induced by Emodin in the Human Neuroblastoma SH-SY5Y Cells Exposed to Hydrogen Peroxide: Involvement of the AMPK/Nrf2 Signaling Pathway.

Emodin (EM; 1,3,8-trihydroxy-6-methylanthracene-9,10-dione; C15H10O5) is an anthraquinone and exerts cytoprotective effects, as observed in both in vitro and in vivo experimental models. Mitochondrial dysfunction induced by reactive species plays a central role in the onset and progression of different human diseases. Thus, we have tested here whether a pretreatment (for 4 h) with EM (at 40 µM) would be able to promote mitochondrial protection in the human neuroblastoma SH-SY5Y cells exposed to the pro-oxidant agent hydrogen peroxide (H2O2). We found that the pretreatment with EM suppressed the effects of H2O2 on the activity of the mitochondrial complexes I and V, as well as on the production of adenosine triphosphate (ATP) and on the mitochondrial membrane potential (MMP). EM also prevented the H2O2-induced collapse in the tricarboxylic acid cycle (TCA) function. An anti-inflammatory role for EM was also observed in this experimental model, since this anthraquinone decreased the secretion of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) by the H2O2-challenged cells. Inhibition of the adenosine monophosphate-activated protein kinase (AMPK) or silencing of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) abolished the protection induced by EM in the H2O2-treated cells. Therefore, EM prevented the H2O2-induced mitochondrial dysfunction and pro-inflammatory state in the SH-SY5Y cells by an AMPK/Nrf2-dependent manner.