Nicotinamide Adenine Dinucleotide Precursor Supplementation Modulates Neurite Complexity and Survival in Motor Neurons from Amyotrophic Lateral Sclerosis Models.

Aims: Increasing nicotinamide adenine dinucleotide (NAD+) availability has been proposed as a therapeutic approach to prevent neurodegeneration in amyotrophic lateral sclerosis (ALS). Accordingly, NAD+ precursor supplementation appears to exert neuroprotective effects in ALS patients and mouse models. The mechanisms mediating neuroprotection remain uncertain but could involve changes in multiple cell types. We investigated a potential direct effect of the NAD+ precursor nicotinamide mononucleotide (NMN) on the health of cultured induced pluripotent stem cell (iPSC)-derived human motor neurons and in motor neurons isolated from two ALS mouse models, that is, mice overexpressing wild-type transactive response DNA binding protein-43 (TDP-43) or the ALS-linked human superoxide dismutase 1 with the G93A mutation (hSOD1G93A). Results: NMN treatment increased the complexity of neuronal processes in motor neurons isolated from both mouse models and in iPSC-derived human motor neurons. In addition, NMN prevented neuronal death induced by trophic factor deprivation. In mouse and human motor neurons expressing ALS-linked mutant superoxide dismutase 1, NMN induced an increase in glutathione levels, but this effect was not observed in nontransgenic or TDP-43 overexpressing motor neurons. In contrast, NMN treatment normalized the TDP-43 cytoplasmic mislocalization induced by its overexpression. Innovation: NMN can directly act on motor neurons to increase the growth and complexity of neuronal processes and prevent the death induced by trophic factor deprivation. Conclusion: Our results support a direct beneficial effect of NAD+ precursor supplementation on the maintenance of the neuritic arbor in motor neurons. Importantly, this was observed in motor neurons isolated from two different ALS models, with and without involvement of TDP-43 pathology, supporting its therapeutic potential in sporadic and familial ALS.

FABP7 drives an inflammatory response in human astrocytes and is upregulated in Alzheimer's disease.

Alzheimer's disease (AD), the most common cause of dementia in the elderly, is characterized by the accumulation of intracellular neurofibrillary tangles, extracellular amyloid plaques, and neuroinflammation. In partnership with microglial cells, astrocytes are key players in the regulation of neuroinflammation. Fatty acid binding protein 7 (FABP7) belongs to a family of conserved proteins that regulate lipid metabolism, energy homeostasis, and inflammation. FABP7 expression is largely restricted to astrocytes and radial glia-like cells in the adult central nervous system. We observed that treatment of primary hippocampal astrocyte cultures with amyloid ß fragment 25-35 (Aß25-35) induces FABP7 upregulation. In addition, FABP7 expression is upregulated in the brain of APP/PS1 mice, a widely used AD mouse model. Co-immunostaining with specific astrocyte markers revealed increased FABP7 expression in astrocytes. Moreover, astrocytes surrounding amyloid plaques displayed increased FABP7 staining when compared to non-plaque-associated astrocytes. A similar result was obtained in the brain of AD patients. Whole transcriptome RNA sequencing analysis of human astrocytes differentiated from induced pluripotent stem cells (i-astrocytes) overexpressing FABP7 identified 500 transcripts with at least a 2-fold change in expression. Gene Ontology enrichment analysis identified (i) positive regulation of cytokine production and (ii) inflammatory response as the top two statistically significant overrepresented biological processes. We confirmed that wild-type FABP7 overexpression induces an NF-κB-driven inflammatory response in human i-astrocytes. On the other hand, the expression of a ligand-binding impaired mutant FABP7 did not induce NF-κB activation. Together, our results suggest that the upregulation of FABP7 in astrocytes could contribute to the neuroinflammation observed in AD.

Production of a potential multistrain probiotic in co-culture conditions using agro-industrial by-products-based medium for fish nutrition.

BACKGROUND: Probiotics are viable microorganisms that when administered in adequate amounts confer health benefits to the host. In fish, probiotic administration has improved growth, and immunological parameters. For this reason, it is necessary production of probiotic bacteria, however, commercial culture mediums used for probiotic growth are expensive, so the design of a "low" cost culture medium is necessary. Therefore, this research aimed to produce a potential multistrain probiotic preparation composed of L. lactis A12 and Priestia species isolated from Nile tilapia (Oreochromis niloticus) gut using an agro-industrial by-products-based culture medium. RESULTS: A Box-Behnken design with three factors (whey, molasses, and yeast extract concentration) was used. As the main results, a high concentration of three components enhanced the viability of L. lactis A12, however, viable cell counts of Priestia species were achieved at low molasses concentrations. The Optimal conditions were 1.00% w/v whey, 0.50% w/v molasses, and 1.50% w/v yeast extract. L. lactis A12 and Priestia species viable counts were 9.43 and 6.89 Log10 CFU/mL, respectively. L. lactis A12 concentration was higher (p < 0.05) in the proposed medium compared to commercial broth. CONCLUSIONS: It was possible to produce L. lactis A12 and Priestia species in co-culture conditions. Whey and molasses were suitable components to produce the multistrain preparation. The cost of the proposed culture medium was 77.54% cheaper than the commercial medium. The proposed culture medium could be an alternative to commercial mediums for the production of this multistrain probiotic.

Nicotinamide Adenine Dinucleotide (NAD+)-Dependent Signaling in Neurological Disorders.

Significance: Nicotinamide adenine dinucleotide (NAD+) participates in redox reactions and NAD+-dependent signaling processes, which couples the enzymatic degradation of NAD+ to posttranslational modifications of proteins or the production of second messengers. Cellular NAD+ levels are dynamically controlled by synthesis and degradation, and dysregulation of this balance has been associated with acute and chronic neuronal dysfunction. Recent Advances: A decline in NAD+ has been observed during normal aging and since aging is the primary risk factor for many neurological disorders, NAD+ metabolism has become a promising therapeutic target and prolific research field in recent years. Critical Issues: In many neurological disorders, either as a primary feature or as consequence of the pathological process, neuronal damage is accompanied by dysregulated mitochondrial homeostasis, oxidative stress, or metabolic reprogramming. Modulating NAD+ availability appears to have a protective effect against such changes observed in acute neuronal damage and age-related neurological disorders. Such beneficial effects could be, at least in part, due to the activation of NAD+-dependent signaling processes. Future Directions: While in many instances the protective effect has been ascribed to the activation of sirtuins, approaches that directly test the role of sirtuins or that target the NAD+ pool in a cell-type-specific manner may be able to provide further mechanistic insight. Likewise, these approaches may afford greater efficacy to strategies aimed at harnessing the therapeutic potential of NAD+-dependent signaling in neurological disorders. Antioxid. Redox Signal. 39, 1150-1166.

Redrawing Cities with Children and Adolescents: Development of a Framework and Opportunity Index for Wellbeing-The REDibuja Study Protocol.

Global changes require urgent integration of health and wellbeing into all urban policies. Complex social and environmental factors define wellbeing outcomes and inequities present in cities. Additionally, political decisions are seldom thought and developed considering the needs and participation of children and adolescents. The REDibuja study aims to develop a multidimensional framework of wellbeing for children and adolescents and to validate an index of opportunities for better wellbeing for children and adolescents in the urban context of Temuco, Chile. This child-centered and cross-sectional study will involve mixed methodologies throughout the implementation of five work packages for two years (2022-2023): (1) development of a conceptual framework for child and adolescent wellbeing, (2) integration of available and public data, (3) studies in the local context, (4) data integration using geographic information systems, and (5) validation of the wellbeing opportunity index for children and adolescents. REDibuja will implement methodologies that until now are little used to facilitate political decisions in our regional context. This process and results could be transferred for assessment and decision-making in Latin America and low- and middle-income countries in other regions.

NR1D1 downregulation in astrocytes induces a phenotype that is detrimental to cocultured motor neurons.

Nuclear receptor subfamily 1 group D member 1 (NR1D1, also known as Rev-erbα) is a nuclear transcription factor that is part of the molecular clock encoding circadian rhythms and may link daily rhythms with metabolism and inflammation. NR1D1, unlike most nuclear receptors, lacks a ligand-dependent activation function domain 2 and is a constitutive transcriptional repressor. Amyotrophic lateral sclerosis (ALS) is the most common adult-onset motor neuron disease, caused by the progressive degeneration of motor neurons in the spinal cord, brain stem, and motor cortex. Approximately 10%-20% of familial ALS is caused by a toxic gain-of-function induced by mutations of the Cu/Zn superoxide dismutase (SOD1). Dysregulated clock and clock-controlled gene expression occur in multiple tissues from mutant hSOD1-linked ALS mouse models. Here we explore NR1D1 dysregulation in the spinal cord of ALS mouse models and its consequences on astrocyte-motor neuron interaction. NR1D1 protein and mRNA expression are significantly downregulated in the spinal cord of symptomatic mice expressing mutant hSOD1, while no changes were observed in age-matched animals overexpressing wild-type hSOD1. In addition, NR1D1 downregulation in primary astrocyte cultures induces a pro-inflammatory phenotype and decreases the survival of cocultured motor neurons. NR1D1 orchestrates the cross talk between physiological pathways identified to be disrupted in ALS (e.g., metabolism, inflammation, redox homeostasis, and circadian rhythms) and we observed that downregulation of NR1D1 alters astrocyte-motor neuron interaction. Our results suggest that NR1D1 could be a potential therapeutic target to prevent astrocyte-mediated motor neuron toxicity in ALS.

Morphological characterization of hemocytes of the brown mussel Perna perna: An update.

Considering the importance of hemocyte characterization for immunological studies, this work aimed to characterize the hemocyte types of Perna perna mussels combining transmission electron microscopy and flow cytometry with the classical optical microscopy. The results indicated four type of hemocytes: hyalinocytes, semigranulocytes, granulocytes and blast-like cells.

Associations Between Movement Behaviors and Emotional Changes in Toddlers and Preschoolers During Early Stages of the COVID-19 Pandemic in Chile.

Background: There is limited evidence about emotional and behavioral responses in toddlers and preschoolers during the novel coronavirus (COVID-19) pandemic, particularly in Latin America. Objective: To assess associations between changes in movement behaviors (physical activity, screen time and sleeping) and emotional changes in toddlers and preschoolers during early stages of the pandemic in Chile. Methods: A cross-sectional study conducted from March 30th to April 27th, 2020. Main caregivers of 1- to 5-year-old children living in Chile answered an online survey that included questions about sociodemographic characteristics, changes in the child's emotions and behaviors, movement behaviors and caregivers' stress during the pandemic. Multiple linear regressions were used to assess the association between different factors and emotional changes in toddlers and preschoolers. Results: In total, 1727 caregivers provided complete data on emotional changes for children aged 2.9 ± 1.36 years old, 47.9% girls. A large proportion of toddlers and preschoolers in Chile experienced emotional and behavioral changes. Most caregivers reported that children "were more affectionate" (78.9%), "more restless" (65.1%), and 'more frustrated' (54.1%) compared with pre-pandemic times. Apart from changes in movement behaviors, factors such as child age, caregivers' age and stress, and residential area (urban/rural) were consistently associated with changes in emotions and behaviors. Conclusion: The pandemic substantially affected the emotions and behaviors of toddlers and preschoolers in Chile. The findings suggest that supportive actions for caregivers may have a positive impact not only on adults but also on children. Mental health promotion programs should consider multilevel approaches in which the promotion of movement behaviors and support for caregivers should be essential pieces for future responses.

Comparison between self-reported and accelerometer-derived measurements for classifying children and adolescents as physically active in Chile.

The aim of this study was to compare self-reported with two accelerometer-derived methods to classify Chilean children and adolescents as physically active. In total, 247 students wore an accelerometer on their hips during 7 consecutive days to classify them as physically active based on (1) daily accumulation of ≥ 60 minutes of moderate-to-vigorous physical activity (MVPA) on each of the seven days, and (2) average MVPA ≥ 60 minutes/day. Also, participants were classified as physically active if they reported being active for at least 60 minutes in all seven days. When using the accelerometer data, 0.8% were active in all seven days, while 10.5% recorded ≥ 60 minutes MVPA per day on average. Based on self-report, 7.2% were physically active. The agreement between self-reported and accelerometer estimations were poor. Important differences were observed between the self-reported and device-derived methods for classifying children and adolescents as physically active. When comparing them, some considerations should be taken. The findings suggest that these methods are not interchangeable. Therefore, if possible, they should be used as complementary measurements.

Altered expression of clock and clock-controlled genes in a hSOD1-linked amyotrophic lateral sclerosis mouse model.

Most physiological processes in mammals are subjected to daily oscillations that are governed by a circadian system. The circadian rhythm orchestrates metabolic pathways in a time-dependent manner and loss of circadian timekeeping has been associated with cellular and system-wide alterations in metabolism, redox homeostasis, and inflammation. Here, we investigated the expression of clock and clock-controlled genes in multiple tissues (suprachiasmatic nucleus, spinal cord, gastrocnemius muscle, and liver) from mutant hSOD1-linked amyotrophic lateral sclerosis (ALS) mouse models. We identified tissue-specific changes in the relative expression, as well as altered daily expression patterns, of clock genes, sirtuins (Sirt1, Sirt3, and Sirt6), metabolic enzymes (Pfkfb3, Cpt1, and Nampt), and redox regulators (Nrf2, G6pd, and Pgd). In addition, astrocytes transdifferentiated from induced pluripotent stem cells from SOD1-linked and FUS RNA binding protein-linked ALS patients also displayed altered expression of clock genes. Overall, our results raise the possibility of disrupted cross-talk between the suprachiasmatic nucleus and peripheral tissues in hSOD1G93A mice, preventing proper peripheral clock regulation and synchronization. Since these changes were observed in symptomatic mice, it remains unclear whether this dysregulation directly drives or it is a consequence of the degenerative process. However, because metabolism and redox homeostasis are intimately entangled with circadian rhythms, our data suggest that altered expression of clock genes may contribute to metabolic and redox impairment in ALS. Since circadian dyssynchrony can be rescued, these results provide the groundwork for potential disease-modifying interventions.

Comparison between self-reported and accelerometer-derived measurements for classifying children and adolescents as physically active in Chile / Comparación entre medidas autoreportadas y provenientes de acelerómetro para clasificar a los niños, niñas y adolescentes como físicamente activos en Chile / Comparação de autorrelato e medidas derivadas de acelerômetro para classificar a atividade física em crianças e adolescentes chilenos

The aim of this study was to compare self-reported with two accelerometer-derived methods to classify Chilean children and adolescents as physically active. In total, 247 students wore an accelerometer on their hips during 7 consecutive days to classify them as physically active based on (1) daily accumulation of ≥ 60 minutes of moderate-to-vigorous physical activity (MVPA) on each of the seven days, and (2) average MVPA ≥ 60 minutes/day. Also, participants were classified as physically active if they reported being active for at least 60 minutes in all seven days. When using the accelerometer data, 0.8% were active in all seven days, while 10.5% recorded ≥ 60 minutes MVPA per day on average. Based on self-report, 7.2% were physically active. The agreement between self-reported and accelerometer estimations were poor. Important differences were observed between the self-reported and device-derived methods for classifying children and adolescents as physically active. When comparing them, some considerations should be taken. The findings suggest that these methods are not interchangeable. Therefore, if possible, they should be used as complementary measurements.

El objetivo fue comparar el autoreporte con dos métodos provenientes de acelerómetría para clasificar a niños, niñas y adolescentes chilenos como físicamente activos. Doscientos cuarenta y siete estudiantes llevaron un acelerómetro en la cintura durante siete días consecutivos y fueron clasificados como físicamente activos, basados en: (1) acumulación diaria de ≥ 60 minutos de actividad física de moderada a vigorosa (AFMV) en cada uno de los siete días, y (2) promedio de AFMV ≥ 60 minutos/día. Asimismo, los participantes fueron clasificados como físicamente activos si reportaron ser activos por al menos 60 minutos en cada uno de los siete días. Al usar los datos del acelerómetro, 0,8% fueron activos los siete días, mientras que un 10,5% registró ≥ 60 minutos AFMV por día promedio. Un 7,2% fue físicamente activos según el autoreporte. El acuerdo entre las estimaciones autoreportadas y el acelerómetro fue pobre. Se observaron importantes diferencias entre los autoreportes y los métodos derivados del dispositivo para clasificar niños, niñas y adolescentes como físicamente activos. Cuando se comparen datos derivados de los acelerómetros y autoreportes esto se debería considerar. Los resultados sugieren que estos métodos no son intercambiables. Por ello, en la medida de lo posible, se deberían usar como medidas complementarias.

O objetivo foi comparar o autorrelato com dois métodos derivados de acelerômetro para classificar crianças e adolescentes chilenos como fisicamente ativas ou inativas. Um total de 247 alunos usaram um acelerômetro no quadril durante sete dias consecutivos e foram classificados como fisicamente ativos com base em: (1) acúmulo diário de ≥ 60 minutos de atividade física de moderada a vigorosa intensidade (AFMV) em cada um dos sete dias e (2) AFMV média por dia de ≥ 60 minutos. Além disso, os participantes foram classificados como fisicamente ativos com base no autorrelato de ser ativo por pelo menos 60 minutos em todos os sete dias. Com o uso dos dados de acelerômetro, 0,8% foram classificados como ativos em todos os sete dias, enquanto 10,5% registravam ≥ 60 minutos de AFMV por dia em média e 7,2% eram fisicamente ativos com base no autorrelato. A concordância foi baixa entre o autorrelato e as estimativas por acelerômetro. Foram observadas diferenças importantes entre o autorrelato e os métodos baseados em dispositivos para classificar as crianças e adolescentes como fisicamente ativos. Algumas considerações são relevantes ao comparar estimativas baseadas em acelerômetro e o autorrelato de atividade física. Os achados sugerem que esses métodos não são intercambiáveis. Portanto, quando possível, devem ser usados como medidas complementares.

STXBP2-R190C Variant in a Patient With Neonatal Hemophagocytic Lymphohistiocytosis (HLH) and G6PD Deficiency Reveals a Critical Role of STXBP2 Domain 2 on Granule Exocytosis.

Neonatal hemophagocytic lymphohistiocytosis (HLH) is a medical emergency that can be associated with significant morbidity and mortality. Often these patients present with familial HLH (f-HLH), which is caused by gene mutations interfering with the cytolytic pathway of cytotoxic T-lymphocytes (CTLs) and natural killer cells. Here we describe a male newborn who met the HLH diagnostic criteria, presented with profound cholestasis, and carried a maternally inherited heterozygous mutation in syntaxin-binding protein-2 [STXBP2, c.568C>T (p.Arg190Cys)] in addition to a severe pathogenic variant in glucose 6-phosphate dehydrogenase [G6PD, hemizygous c.1153T>C (Cys385Arg)]. Although mutations in STXBP2 gene are associated with f-HLH type 5, the clinical and biological relevance of the p.Arg190Cys mutation identified in this patient was uncertain. To assess its role in disease pathogenesis, we performed functional assays and biochemical and microscopic studies. We found that p.Arg190Cys mutation did not alter the expression or subcellular localization of STXBP2 or STX11, neither impaired the STXBP2/STX11 interaction. In contrast, forced expression of the mutated protein into normal CTLs strongly inhibited degranulation and reduced the cytolytic activity outcompeting the effect of endogenous wild-type STXBP2. Interestingly, arginine 190 is located in a structurally conserved region of STXBP2 where other f-HLH-5 mutations have been identified. Collectively, data strongly suggest that STXBP2-R190C is a deleterious variant that may act in a dominant-negative manner by probably stabilizing non-productive interactions between STXBP2/STX11 complex and other still unknown factors such as the membrane surface or Munc13-4 protein and thus impairing the release of cytolytic granules. In addition to the contribution of STXBP2-R190C to f-HLH, the accompanied G6PD mutation may have compounded the clinical symptoms; however, the extent by which G6PD deficiency has contributed to HLH in our patient remains unclear.

Compliance of the 24-Hour Movement Guidelines in 9- to 11-Year-Old Children From a Low-Income Town in Chile.

BACKGROUND: The purpose was to determine the proportion of 9- to 11-year-old children meeting the 24-hour movement guidelines (24-HMG) in a low-income town from Chile. METHODS: Physical activity, sedentary behavior (recreational screen), and sleep times were measured with both questionnaire and accelerometer in 258 children from third to sixth grade. Meeting the 24-HMG was defined as having ≥60 minutes per day of moderate to vigorous physical activity, ≤2 hour day of screen time, and 9 to 11 hours of sleep per night. Compliance rates were calculated as self-reported 24-HMG, with all estimations based on questionnaires, and mixed 24-HMG, in which physical activity and sleep were determined with an accelerometer and sedentary behavior was determined with a questionnaire. RESULTS: About 198 children (10.1 [0.8] y, range 9-11 y) provided valid data for estimating self-reported 24-HMG, and 141 for mixed 24-HMG. Only 3.2% and 0.7% met the 24-HMG when using the self-reported and mixed methods, respectively. When assessing individual recommendations, 13.1% and 3.7% of the sample were physically active based on the self-report and accelerometer, respectively. About a quarter met the sedentary behavior recommendations, while around 50% met the sleep recommendations with both self-reported and mixed methods. CONCLUSIONS: An extremely low percentage of the participants met the 24-HMG. Multicomponent initiatives must be implemented to promote healthy movement behaviors in Chilean children.

Effects of RAGE inhibition on the progression of the disease in hSOD1G93A ALS mice.

Astrocytes play a key role in the progression of amyotrophic lateral sclerosis (ALS) by actively inducing the degeneration of motor neurons. Motor neurons isolated from receptor for advanced glycation end products (RAGE)-knockout mice are resistant to the neurotoxic signal derived from ALS-astrocytes. Here, we confirmed that in a co-culture model, the neuronal death induced by astrocytes over-expressing the ALS-linked mutant hSOD1G93A is prevented by the addition of the RAGE inhibitors FPS-ZM1 or RAP. These inhibitors also prevented the motor neuron death induced by spinal cord extracts from symptomatic hSOD1G93A mice. To evaluate the relevance of this neurotoxic mechanism in ALS pathology, we assessed the therapeutic potential of FPS-ZM1 in hSOD1G93A mice. FPS-ZM1 treatment significantly improved hind-limb grip strength in hSOD1G93A mice during the progression of the disease, reduced the expression of atrophy markers in the gastrocnemius muscle, improved the survival of large motor neurons, and reduced gliosis in the ventral horn of the spinal cord. However, we did not observe a statistically significant effect of the drug in symptoms onset nor in the survival of hSOD1G93A mice. Maintenance of hind-limb grip strength was also observed in hSOD1G93A mice with RAGE haploinsufficiency [hSOD1G93A ;RAGE(+/-)], further supporting the beneficial effect of RAGE inhibition on muscle function. However, no benefits were observed after complete RAGE ablation. Moreover, genetic RAGE ablation significantly shortened the median survival of hSOD1G93A mice. These results indicate that the advance of new therapies targeting RAGE in ALS demands a better understanding of its physiological role in a cell type/tissue-specific context.

FABP7 upregulation induces a neurotoxic phenotype in astrocytes.

Fatty acid binding proteins (FABPs) are key regulators of lipid metabolism, energy homeostasis, and inflammation. They participate in fatty acid metabolism by regulating their uptake, transport, and availability of ligands to nuclear receptors. In the adult brain, FABP7 is especially abundant in astrocytes that are rich in cytoplasmic granules originated from damaged mitochondria. Mitochondrial dysfunction and oxidative stress have been implicated in the neurodegenerative process observed in amyotrophic lateral sclerosis (ALS), either as a primary cause or as a secondary component of the pathogenic process. Here we investigated the expression of FABP7 in animal models of human superoxide dismutase 1 (hSOD1)-linked ALS. In the spinal cord of symptomatic mutant hSOD1-expressing mice, FABP7 is upregulated in gray matter astrocytes. Using a coculture model, we examined the effect of increased FABP7 expression in astrocyte-motor neuron interaction. Our data show that FABP7 overexpression directly promotes an NF-κB-driven pro-inflammatory response in nontransgenic astrocytes that ultimately is detrimental for motor neuron survival. Addition of trophic factors, capable of supporting motor neuron survival in pure cultures, did not prevent motor neuron loss in cocultures with FABP7 overexpressing astrocytes. In addition, astrocyte cultures obtained from symptomatic hSOD1-expressing mice display upregulated FABP7 expression. Silencing endogenous FABP7 in these cultures decreases the expression of inflammatory markers and their toxicity toward cocultured motor neurons. Our results identify a key role of FABP7 in the regulation of the inflammatory response in astrocytes and identify FABP7 as a potential therapeutic target to prevent astrocyte-mediated motor neuron toxicity in ALS.

Single and repeated low-dose UVB radiation exposures affect the visual system.

The visual system is an important biological indicator of effects induced by ultraviolet (UV) radiation. However, research has extensively investigated the effects of high-dose UV radiation in a single exposure, thus, the differential of this work was to investigate the effects of UVB radiation in low doses in single and repeated exposure. Therefore, we investigated the effects of repeated exposure to environmental UVB doses (0.09 J/cm2) on the retina and optic lobes of the crab Neohelice granulata. We evaluated the reactive oxygen species (ROS) concentration, antioxidant capacity against peroxyl radicals (ACAP) levels, catalase (CAT) and glutathione S-transferase (GST) activities and lipoperoxidation (LPO) levels and performed histological analysis. The crabs were exposed to UVB radiation for 1 or 60 days, while the control group was exposed to visible light. In the retina region, increases in ROS concentration and CAT and GST activities after the single exposure were observed. After 60 days of exposure, we observed an increase in ACAP levels. In the optic lobes, we observed an increase in GST activity and a decrease in LPO levels after the single exposure. However, we observed an increase in ROS concentration after 60 days of exposure. Moreover, after 60 days of exposure, infiltrating hemocytes in the retina and disorganization in neuron cell bodies of the external medulla were observed. In this sense, single and repeated exposure to low doses of UVB radiation induced changes in oxidative status and inflammatory process in the visual system of the crab Neohelice granulata.

Effects of DCOIT (4,5-dichloro-2-octyl-4-isothiazolin-3-one) to the haemocytes of mussels Perna perna.

Bivalve molluscs rely only on an innate immune system to execute cellular and humoral processes. Haemocytes, the haemolymph circulating cells, play a major role in this type of immunity, principally regarding cellular defences. Considering that environmental pollutants can affect the immune system of invertebrates, this work evaluated the effects of the antifouling biocide 4,5-dicloro-2-n-octil-4-isotiazolin-3-ona (DCOIT) on the haemocytes of mussels Perna perna. Individuals were exposed to 0 (control), 0.1 µg L-1 and 10 µg L-1 of DCOIT for up to 96 h. The analysed parameters included: total (THC) and differential (DHC) haemocyte count, cellular viability, adhesion capacity, phagocytic activity, levels of reactive oxygen species and DNA damage. Moreover, the stress on stress (SOS) response of mussels was analysed as a general stress index. The results show that DCOIT increased the haemocyte adhesion capacity and caused a decrease in THC and in the haemocyte viability after 24 h of exposure. After 96 h of exposure, DCOIT only affected the haemocyte adhesion capacity, which was decreased by biocide exposure. Moreover, exposure to DCOIT for 96 h did not affect the capacity for air survival of mussels. These results indicate that DCOIT interferes in important parameters associated with the innate immunity of P. perna, mainly after 24 h of exposure. It is suggested that the animals were able to develop some compensatory response strategy, making them more resistant to the biocide.

Evaluation of the NAD+ biosynthetic pathway in ALS patients and effect of modulating NAD+ levels in hSOD1-linked ALS mouse models.

Amyotrophic lateral sclerosis (ALS) is characterized by progressive degeneration of motor neurons. Astrocytes from diverse ALS models induce motor neuron death in co-culture. Enhancing NAD+ availability, or increasing the expression of the NAD+-dependent deacylases SIRT3 and SIRT6, abrogates their neurotoxicity in cell culture models. To determine the effect of increasing NAD+ availability in ALS mouse models we used two strategies, ablation of a NAD+-consuming enzyme (CD38) and supplementation with a bioavailable NAD+ precursor (nicotinamide riboside, NR). Deletion of CD38 had no effect in the survival of two hSOD1-linked ALS mouse models. On the other hand, NR-supplementation delayed motor neuron degeneration, decreased markers of neuroinflammation in the spinal cord, appeared to modify muscle metabolism and modestly increased the survival of hSOD1G93A mice. In addition, we found altered expression of enzymes involved in NAD+ synthesis (NAMPT and NMNAT2) and decreased SIRT6 expression in the spinal cord of ALS patients, suggesting deficits of this neuroprotective pathway in the human pathology. Our data denotes the therapeutic potential of increasing NAD+ levels in ALS. Moreover, the results indicate that the approach used to enhance NAD+ levels critically defines the biological outcome in ALS models, suggesting that boosting NAD+ levels with the use of bioavailable precursors would be the preferred therapeutic strategy for ALS.

Oxidative stress: Noradrenaline as an integrator of responses in the neuroendocrine and immune systems of the ascidian Phallusia nigra.

Neurotransmitters play key roles in regulating the homeostasis of organisms in stressful environments. Noradrenaline (NA) is the main neurotransmitter known to modulate immunological parameters, and is important in the crosstalk between the neuroendocrine and immune systems. In this study, using the ascidian Phallusia nigra, we analyzed the level of catecholamines (CA) in the plasma after mechanical stress, and the effect of NA on the oxidative stress (OS) displayed by immune cells. We measured the concentration of reactive oxygen species (ROS), and analyzed whether α- and/or ß-adrenoreceptors (ARs) are involved in ROS modulation, lipid peroxidation (LPO), antioxidant capacity against peroxyl radicals (ACAP), and activity of the enzymes catalase (CAT) and glutathione S transferase (GST) in immune cells after incubation with different concentrations of NA, with or without zymosan (ZnA) challenge. The results showed that NA reduced ROS production, even in immune cells challenged with ZnA, and that this modulation occurred through α1-and ß1-ARs. ACAP levels showed different responses, depending on whether immune cells were challenged or not with ZnA, and also depending on the NA concentration: 1.0 µM NA increased ACAP levels, but 10.0 µM reduced ACAP levels. NA enhanced the activity of CAT and GST in ZnA-challenged and non-challenged immune cells, while 1.0 and 10.0 µM NA effectively reduced LPO. Taken together, these results show that NA can protect cells from ROS damage, decreasing ROS production and LPO, and enhancing ACAP as well as the activity of CAT and GST. The approach used here with this model contributes to understanding the relationship between the neuroendocrine and immune systems, revealing new effects of NA on OS regulation in ascidians.