Renal profile of patients treated with elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide fumarate and dolutegravir/abacavir/lamivudine: 120-week results from a real-world cohort.

Elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide fumarate (EVG/c/FTC/TAF) and dolutegravir/abacavir/lamivudine (DTG/ABC/3TC) are currently available for HIV patients. OBJECTIVES: This study evaluated modifications in the renal safety profile in a large real-world cohort of patients who had received EVG/c/FTC/TAF or DTG/ABC/3TC. METHODS: A retrospective observational study of HIV-infected patients who received EVG/c/FTC/TAF or DTG/ABC/3TC between March 2015 and June 2019 at a reference hospital in north-western Spain was conducted. Epidemiological, clinical, immunovirological data and information regarding antiretroviral therapy were recorded. The statistical differences between treatments were calculated. RESULTS: A total of 457 patients were evaluated, 266 using EVG/c/FTC/TAF and 191 using DTG/ABC/3TC. Up to week 120, serum creatinine improved in both study groups among experienced patients (EVG/c/FTC/TAF 1.01±0.24 vs 0.91±0.19, p<0.001; DTG/ABC/3TC 1.08±0.24 vs 1.02±0.31, p<0.001), while in naïve patients serum creatinine remained stable compared with baseline. Statistically significant differences were found in serum creatinine when comparing both treatments at week 48 in experienced (0.94±0.21 vs 1.09±0.28, p<0.001) and naïve patients (0.89±0.16 vs 1.06±0.20, p=0.001), and among experienced patients at week 120 (0.91±0.19 vs 1.02±0.31, p=0.015) for the EVG/c/FTC/TAF and DTG/ABC/3TC groups, respectively. During the follow-up, 39 patients in EVG/c/FTC/TAF and 33 in DTG/ABC/3TC (p=0.449) discontinued treatment. The main reason for stopping treatment was adverse events, which were similar in both groups. CONCLUSIONS: During the follow-up, patients experienced changes that were not clinically relevant in both treatment groups. Differences in renal events were not found.

Discontinuation due to neuropsychiatric adverse events with efavirenz- and dolutegravir-based antiretroviral therapy: a comparative real-life study.

OBJECTIVES: Despite the high efficacy of antiretroviral treatment, no drug is free from adverse events (AEs). Efavirenz (EFV) and dolutegravir (DTG) are antiretroviral drugs for which neuropsychiatric adverse events (NPAEs) have been described. This study evaluated the safety and tolerability of DTG-based and EFV-based antiretroviral regimens in HIV-infected patients. METHODS: A retrospective observational study was carried out in HIV-infected patients who started DTG- or EFV-based antiretroviral treatment from January 2008 to December 2018 at a reference hospital in north-western Spain. Epidemiological, clinical and immunovirological data were recorded. A statistical analysis was performed with SPSS software. RESULTS: A total of 282 DTG- and 148 EFV-based therapies were initiated. During follow-up, statistically significant differences have been found between the rate of patients who discontinued DTG and EFV due to AEs (12.1% vs 35.8%, p<0.001) and the main AEs in both groups, NPAEs (8.2% vs 25.0%, p<0.001). Female gender (OR 2.610 (95% CI 1.327 to 5.133), p=0.005) was associated with discontinuations due to AEs. Patients with documented psychiatric disorders were at higher risk of discontinuation due to NPAEs (OR 4.782 (95% CI 1.190 to 19.220), p=0.027). The multivariate analysis showed a 61.2% risk reduction in benzodiazepine prescriptions in patients treated with DTG. In both groups, patients needed consultation and follow-up in the psychiatry unit (16.9% in the EFV group and 8.9% in the DTG group, p=0.021). CONCLUSIONS: We found a high rate of discontinuations due to AEs and NPAEs, prescription of benzodiazepines and a requirement for consultation in a psychiatric unit in both treatment groups, especially with EFV.

Does the Rural Environment Influence Symptomatology and Optimize the Effectiveness of Disease Acceptance? A Study Among Women With Fibromyalgia.

The present study aims to explore whether the symptoms associated with fibromyalgia are contextually influenced by the area of residence (rural/urban). Furthermore, it is analyzed whether the effect of the acceptance of the disease on the emotional, cognitive and physical symptoms is moderated by the patients' place of residence. Using a cross-sectional design, a total of 234 women with fibromyalgia (mean age = 56.91 years; SD = 8.94) were surveyed, of which 55.13% resided in rural areas and 44.87% in urban areas. Self-reported questionnaires were used to assess pain severity, anxiety and depression, functional limitation, physical and mental fatigue and acceptance of the disease. The results show significant differences in acceptance (p = 0.040), pain (p < 0.001), and physical and mental fatigue (p = 0.003 and p = 0.004, respectively) between patients from rural and urban areas. The rural area patients presented higher levels of acceptance and pain and lesser levels of physical and mental fatigue compared to the urban area. The moderation analysis add that, only in patients from the rural area, the variables of physical symptoms (pain, functional limitation, and physical fatigue) were significantly and negatively associated with acceptance. This study addresses for the first time the role of the place of residence in suffering from fibromyalgia, suggesting that the rural or urban environment plays a relevant role in the severity and/or management of symptoms in fibromyalgia women. Limitations and practical implications are also discussed.

Adicción Alimentaria: ¿Otro determinante asociado a la adhesión del tratamiento nutricional? / Dependência de Alimentos: Outro determinante associado à adesão de tratamento nutricional? / Food Addiction: Another determinant associated with the adhesion of nutritional treatment

Resumen: Los Programas de Prevención Secundaria Cardiovascular se han desarrollado para mejorar el proceso de atención de pacientes con cardiopatía isquémica y/o con alto riesgo cardiovascular. Sin embargo, la adherencia terapéutica en algunos aspectos es limitada, por lo que conocer la prevalencia de adicción alimentaria en esta población podría ser de utilidad para redireccionar la intervención nutricional. El objetivo de este trabajo es estudiar la prevalencia de adicción alimentaria de los pacientes que asisten al Programa de Prevención Secundaria Cardiovascular del Fondo nacional de Recursos en el período octubre a diciembre 2016 en el departamento de Montevideo, Uruguay. Se trata de un estudio descriptivo, observacional y de corte transversal. Se estudian 153 pacientes a los que se les aplicó el cuestionario estructurado YFAS de Gearhardt y valoración del estado nutricional a través del IMC extraído del Sistema Informático María del FNR. El 63% de la población fue del sexo masculino, 8 de cada 10 de los encuestados fueron adultos mayores (≥60 años), y predominó la malnutrición por exceso en el 70% de los encuestados, encontrándose obesidad en 4 de cada 10. La prevalencia de adicción alimentaria en el total de los encuestados fue del 5,9% (IC95%=2,7-10,9), siendo más prevalente en el rango etáreo de menores de 60 años. Dentro de la categoría de obesidad la prevalencia de adicción alimentaria fue 13,8% seguida por el sobrepeso. Se observa en un 10% de la población la presencia de 3 o más síntomas para adicción alimentaria sin tener aún la significación clínica que determine la presencia de la patología, predominando esto en la población del sexo masculino y en los menores de 60 años. Esta presencia dentro de la categoría de obesidad se puede ver en un 10%. Se concluye que casi el 6% de la población presentó adicción alimentaria y que un 10% tuvieron 3 o más síntomas propios de la patología. Incluir el test en la consulta de los pacientes pertenecientes al PPSCV permitiría derivar al equipo de psicología de la institución los pacientes que así lo requieran.

Resumo: Programas secundários de prevenção cardiovascular foram desenvolvidos para melhorar o processo de atendimento de pacientes com cardiopatia isquêmica e / ou com alto risco cardiovascular. No entanto, a adesão terapêutica em alguns aspectos é limitada, portanto, conhecer a prevalência de dependência alimentar nessa população pode ser útil para redirecionar a intervenção nutricional. O objetivo deste trabalho é estudar a prevalência de dependência alimentar de pacientes do Programa de Prevenção Secundária Cardiovascular do Fundo Nacional de Recursos no período de outubro a dezembro de 2016 no departamento de Montevidéu, Uruguai. Trata-se de um estudo descritivo, observacional e transversal. Foram estudados 153 pacientes aos quais foi aplicado o questionário estruturado YFAS de Gearhardt e a avaliação do estado nutricional por meio do IMC extraído do María Informatics System do FNR. 63% da população era do sexo masculino, 8 em cada 10 entrevistados eram adultos mais velhos (≥60 anos) e a supernutrição prevaleceu em 70% dos entrevistados, com obesidade em 4 em cada 10. a prevalência de dependência alimentar no total dos entrevistados foi de 5,9% (IC95% = 2,7-10,9), sendo mais prevalente na faixa etária de crianças menores de 60 anos. Na categoria obesidade, a prevalência de dependência alimentar foi de 13,8%, seguida pelo excesso de peso. Observa-se em 10% da população a presença de 3 ou mais sintomas de dependência alimentar sem ainda ter o significado clínico que determina a presença da patologia, predominantemente na população masculina e naqueles com menos de 60 anos. Essa presença na categoria obesidade pode ser vista em 10%. Conclui-se que quase 6% da população apresentava dependência alimentar e 10% apresentavam 3 ou mais sintomas da patologia. A inclusão do teste na consulta de pacientes pertencentes ao PPSCV permitiria que os pacientes que precisassem dele se referirem à equipe de psicologia da instituição.

Summary: The Cardiovascular Secondary Prevention Programs have been developed to improve the care process for patients with ischemic heart disease and / or with high cardiovascular risk. However, the therapeutic adherence in some aspects is limited, so knowing the prevalence of food addiction in this population could be useful to redirect the nutritional intervention. The objective of this study is to study the prevalence of food addiction in patients attending the Cardiovascular Secondary Prevention Program of the National Resources Fund from October to December 2016 in the department of Montevideo. It is a descriptive, observational and cross-sectional study. 153 patients were studied, the structured YFAS questionnaire of Gearhardt was applied and the nutritional status was assessed through the IMC extracted from the Maria IT System of the FNR. 63% of the population was male, 8 out of 10 of the respondents were elderly (≥60 years), and excess malnutrition predominated in 70% of respondents, with obesity in 4 out of 10. The prevalence of food addiction in the total of the respondents was 5.9% (95% CI = 2.7-10.9), being more prevalent in the age range of those under 60 years of age. Within the category of obesity, the prevalence of food addiction was 13.8%, followed by overweight. The presence of 3 or more symptoms for food addiction was observed in 10% of the population without having the clinical significance that determines the presence of the pathology, predominating in the male population and in those under 60 years of age. This presence within the category of obesity can be seen in 10%. It is concluded that almost 6% of the population had food addiction and 10% had 3 or more symptoms typical of the pathology. Including the test in the appointment of patients belonging to the CSPP would allow referral to the psychology team of the institution of those who require it.

Teleconsultation for the Pharmaceutical Care of HIV Outpatients in Receipt of Home Antiretrovirals Delivery: Clinical, Economic, and Patient-Perceived Quality Analysis.

Background/Introduction:Pharmacist teleconsultations, combined with home drug delivery or mail-order pharmacy (MOP), can help hospital outpatients with difficulties accessing treatment. The objectives of this study are to describe a teleconsultation protocol and to evaluate clinical, economic, and patient-perceived quality results.Materials and Methods:A cohort observational study was carried out for 3 years on HIV outpatients. Clinical variables were adherence, plasma HIV-RNA, and CD4+ levels. A pharmacoeconomic analysis was carried out through a cost-minimization study. Patient-perceived quality was assessed through a satisfaction survey. Simple random sampling was performed for 95% safety, accuracy ±1%, and losses ±20%.Results:The 38 participants (sample size) consisted of 82% male patients, aged 44.7 ± 8.4 years. There were 854 teleconsultations and 100% treatment adherence. All HIV outpatients kept virally suppressed (p = 1.00) and maintained a controlled immunological level (p = 0.87). The economic evaluation revealed 137 ± 23 € patient/year costs-saved and 18.5 ± 7.2 h/patient/year working time gained. Patient-perceived quality average score was >9.4 out of 10 in all items; the most valued factors were the saving of direct costs and reconciliation with work commitments (45%) and the least valued attributes were making the payment for the shipment and having to adjust to a telephone appointment (41%).Discussion/Conclusions:A teleconsultation protocol associated with home antiretrovirals delivery or MOP obtains a high degree of satisfaction from the HIV hospital outpatients receiving treatment, without repercussions on the therapeutic objectives and with the saving of important direct costs for the patient and indirect costs in relation to labor productivity.

Olfactory and cognitive functioning in patients with fibromyalgia.

Fibromyalgia is a complex syndrome characterized by chronic musculoskeletal pain. Previous research has found impaired olfactory function and cognitive deficits in patients with fibromyalgia. In non-clinical population (middle-aged and elderly populations) an association has been found between olfactory function and cognitive impairment. Therefore, olfactory impairment identification could be considered as a reliable and early marker predicting the decline of cognitive function in mild cognitive impairment, Alzheimer's disease and Parkinson's disease. Nevertheless, the relationship between cognitive and olfactory functioning has not been studied in fibromyalgia patients. The aims of the study have been to analyze the cognitive and olfactory functioning and their association in a sample of women with fibromyalgia in comparison to women without a chronic pain disease. A transversal study was conducted with fibromyalgia patients (n = 146) and women without a chronic pain disease (n = 122). Data were collected using the WAIS-III (cognitive functioning) and the CCCRC test (olfactory functioning). Descriptive analysis, Student's t test and chi-square tests were conducted. There were significant differences in perceptive organization and processing speed, and in odour identification, odour threshold and total CCCRC score; the lower scores were for fibromyalgia patients. Among the fibromyalgia patients there were significant differences in the verbal scale, the manipulative scale, the total scale and processing speed between normosmic and hyposmic patients, the lowest scores were for hyposmic patients. Although previous literature has shown that self-reports by fibromyalgia patients inform about an enhanced olfactory acuity, the current study suggests that there are deficits in olfactory functioning in these patients. Also, the relationship between olfactory and cognitive functioning in fibromyalgia patients, due to the lack of previous studies, suggests a new line of research. Taken together, these results suggest that olfactory functioning could be included in a bio-psycho-social approach of fibromyalgia patients, because of the association with cognitive functioning.

Treatment with tenofovir alafenamide fumarate worsens the lipid profile of HIV-infected patients versus treatment with tenofovir disoproxil fumarate, each coformulated with elvitegravir, cobicistat, and emtricitabine.

Two elvitegravir/cobicistat-based therapies combined with emtricitabine/tenofovir disoproxil fumarate (EVG/c/FTC/TDF) or emtricitabine/tenofovir alafenamide fumarate (EVG/c/FTC/TAF) are currently available for HIV patients. This study evaluated the modifications in the lipid profile of patients who received these treatments in the last three years at our institution. A retrospective observational study in HIV-infected patients who received EVG/c/FTC/TDF or EVG/c/FTC/TAF from January 2015 to January 2018 at a reference hospital in northwestern Spain was carried out. Epidemiological, clinical and immunovirological data were recorded. A statistical analysis was performed using SPSS software. A total of 384 EVG/c-based therapies were initiated during the study period, 151 EVG/c/FTC/TDF and 233 EVG/c/FTC/TAF. A significantly negative influence in all the lipid profile parameters in experienced patients and total cholesterol (TC), and LDL-C in naïve patients were observed after 48 weeks of treatment with EVG/c/FTC/TAF, while these parameters remained stable in the EVG/c/FTC/TDF group. During follow-up, a greater proportion of patients had lipid levels above the normal range (63.1% TC, 56.2% LDL-C) and new lipid-modifying drugs were prescribed (11.9%) in the EVG/c/FTC/TAF group. The number of cardiovascular risk factors (OR 1.66 [95% CI 1.01-2.72]; P = 0.043) was recognised as an independent predictor of lipid-lowering prescription for patients treated with both EVG/c/FTC/TDF and EVG/c/FTC/TAF. For patients treated with EVG/c/FTC/TAF, the mean total cholesterol to HDL ratio in the first 48 weeks of the study treatment was associated with a higher likelihood of lipid-lowering prescription in multivariate analysis (OR 1.6 [95% CI 1.12-2.52]; P = 0.011). Significant changes in lipid profile have been observed in patients who have received EVG/c/FTC/TAF. It was necessary to prescribe almost twice the number of lipid-lowering drugs to patients who received EVG/c/FTC/TAF (11.9%) vs EVG/c/FTC/TDF (4.7%).

Attachment styles, pain intensity and emotional variables in women with fibromyalgia.

This study aims to explore the relations between attachment styles and pain intensity and certain emotional variables (anxiety, depression and alexithymia) in a sample of fibromyalgia patients, in comparison with healthy women. Data were collected from 146 women with fibromyalgia and 122 healthy women. The variables studied were attachment style, pain intensity, anxiety, depression and alexithymia dimensions. Patients with fibromyalgia showed lower percentages of secure attachment style (69.9% vs. 86%) whilst showing higher avoidant attachment (19.8% vs. 7.4%), as well as increased numbers of anxious-ambivalent attachment (10.3% vs. 6.6%) than healthy women (X2  = 9.915, p = .007). Also, fibromyalgia patients showed significantly higher scores in two of the insecure attachment factors (p < .000; p = .020) and lower scores on the secure attachment factor (p = .008) in comparison with healthy women. Higher scores of alexithymia were found in women showing anxious-ambivalent and avoidant attachment styles in comparison with those showing a secure attachment style, regardless of the group they belonged to. In fibromyalgia patients, higher anxiety (p = .005) was found among the women with anxious-ambivalent attachment styles (Mean = 15.15; SD = 1.15) in comparison with those with secure attachment style (Mean = 11.18; SD = .45). No relation was found between attachment style and pain intensity. Avoidant attachment seems to carry out a contradictory role and warrants further research. The results found seem to highlight the need for the Attachment-Diathesis Model of Chronic Pain to include attachment styles as a predictor of the emotional experience of pain in fibromyalgia patients.

High prevalence of subtype F in newly diagnosed HIV-1 persons in northwest Spain and evidence for impaired treatment response.

HIV-1 non-B subtype variants were found in 37.8% of 296 newly diagnosed persons in northwest Spain over the past 5 years. Subtype F was the most prevalent non-B subtype (29.6%) and displayed preferential transmission among MSM. Virologic response rates to antiretroviral therapy were lower among F subtypes compared to B subtypes at weeks 24 (31% vs. 78.3%), 48 (51.7% vs. 85.2%), and 96 (61.1% vs. 94.3%) of therapy. Subtype F was independently associated with virological response at 24 weeks.

Thioredoxin reductase 1 deficiency enhances selenite toxicity in cancer cells via a thioredoxin-independent mechanism.

Selenium is an essential trace element in mammals, but is toxic at high levels. It is best known for its cancer prevention activity, but cancer cells are more sensitive to selenite toxicity than normal cells. Since selenite treatment leads to oxidative stress, and the Trx (thioredoxin) system is a major antioxidative system, we examined the interplay between TR1 (Trx reductase 1) and Trx1 deficiencies and selenite toxicity in DT cells, a malignant mouse cell line, and the corresponding parental NIH 3T3 cells. TR1-deficient cells were far more sensitive to selenite toxicity than Trx1-deficient or control cells. In contrast, this effect was not seen in cells treated with hydrogen peroxide, suggesting that the increased sensitivity of TR1 deficiency to selenite was not due to oxidative stress caused by this compound. Further analyses revealed that only TR1-deficient cells manifested strongly enhanced production and secretion of glutathione, which was associated with increased sensitivity of the cells to selenite. The results suggest a new role for TR1 in cancer that is independent of Trx reduction and compensated for by the glutathione system. The results also suggest that the enhanced selenite toxicity of cancer cells and simultaneous inhibition of TR1 can provide a new avenue for cancer therapy.

Selenoprotein S expression in reactive astrocytes following brain injury.

Selenoprotein S (SelS) is an endoplasmic reticulum (ER)-resident protein involved in the unfolded protein response. Besides reducing ER-stress, SelS attenuates inflammation by decreasing pro-inflammatory cytokines. We have recently shown that SelS is responsive to ischemia in cultured astrocytes. To check the possible association of SelS with astrocyte activation, here we investigate the expression of SelS in two models of brain injury: kainic acid (KA) induced excitotoxicity and cortical mechanical lesion. The regulation of SelS and its functional consequences for neuroinflammation, ER-stress, and cell survival were further analyzed using cultured astrocytes from mouse and human. According to our immunofluorescence analysis, SelS expression is prominent in neurons and hardly detectable in astrocytes from control mice. However, brain injury intensely upregulates SelS, specifically in reactive astrocytes. SelS induction by KA was evident at 12 h and faded out after reaching maximum levels at 3-4 days. Analysis of mRNA and protein expression in cultured astrocytes showed SelS upregulation by inflammatory stimuli as well as ER-stress inducers. In turn, siRNA-mediated SelS silencing combined with adenoviral overexpression assays demonstrated that SelS reduces ER-stress markers CHOP and spliced XBP-1, as well as inflammatory cytokines IL-1ß and IL-6 in stimulated astrocytes. SelS overexpression increased astrocyte resistance to ER-stress and inflammatory stimuli. Conversely, SelS suppression compromised astrocyte viability. In summary, our results reveal the upregulation of SelS expression in reactive astrocytes, as well as a new protective role for SelS against inflammation and ER-stress that can be relevant to astrocyte function in the context of inflammatory neuropathologies.

Response of transcription factor NFATc3 to excitotoxic and traumatic brain insults: identification of a subpopulation of reactive astrocytes.

Astrocytes react to brain injury triggering neuroinflammatory processes that determine the degree of neuronal damage. However, the signaling events associated to astrocyte activation remain largely undefined. The nuclear factor of activated T-cells (NFAT) is a transcription factor family implicated in activation of immune cells. We previously characterized the expression of NFAT isoforms in cultured astrocytes, and NFAT activation in response to mechanical lesion. Here we analyze NFATc3 in two mouse models of inflammatory brain damage: hippocampal excitotoxicity induced by intracerebral kainic acid (KA) injection and cortical mechanical lesion. Immunofluorescence results demonstrated that NFATc3 is specifically induced in a subset of reactive astrocytes, and not in microglia or neurons. In KA-treated brains, NFATc3 expression is transient and NFATc3-positive astrocytes concentrate around damaged neurons in areas CA3 and CA1. Complementary Western blot and RT-PCR analysis revealed an NFAT-dependent induction of RCAN1-4 and COX-2 in hippocampus as soon as 6 h after KA exposure, indicating that NFAT activation precedes NFATc3 over-expression. Moreover, activation of NFAT by ATP increased NFATc3 mRNA levels in astrocyte cultures, suggesting that NFATc3 expression is controlled through an auto-regulatory loop. Meanwhile, stab wound enhanced NFATc3 expression specifically in a subclass of reactive astrocytes confined within the proximal layer of the glial scar, and GFAP immunoreactivity was attenuated in NFATc3-expressing astrocytes. In conclusion, our work establishes NFATc3 as a marker of activation for a specific population of astrocytes in response to brain damage, which may have consequences for neuronal survival.

A proteomic analysis of PKCε targets in astrocytes: implications for astrogliosis.

Astrocytes are glial cells in the central nervous system (CNS) that play key roles in brain physiology, controlling processes, such as neurogenesis, brain energy metabolism and synaptic transmission. Recently, immune functions have also been demonstrated in astrocytes, influencing neuronal survival in the course of neuroinflammatory pathologies. In this regard, PKCepsilon (PKCε) is a protein kinase with an outstanding role in inflammation. Our previous findings indicating that PKCε regulates voltage-dependent calcium channels as well as morphological stellation imply that this kinase controls multiple signalling pathways within astrocytes, including those implicated in activation of immune functions. The present study applies proteomics to investigate new protein targets of PKCε in astrocytes. Primary astrocyte cultures infected with an adenovirus that expresses constitutively active PKCε were compared with infection controls. Two-dimensional gel electrophoresis clearly detected 549 spots in cultured astrocytes, and analysis of differential protein expression revealed 18 spots regulated by PKCε. Protein identification by mass spectrometry (nano-LC-ESI-MS/MS) showed that PKCε targets molecules with heterogeneous functions, including chaperones, cytoskeletal components and proteins implicated in metabolism and signalling. These results support the notion that PKCε is involved in astrocyte activation; also suggesting that multiple astrocyte-dependent processes are regulated by PKCε, including those associated to neuroinflammation.

Staging of alpha-synuclein in the olfactory bulb in a model of Parkinson's disease: cell types involved.

Impaired olfaction is an early symptom of Parkinson's disease. The underlying neuropathology likely includes alpha-synucleinopathy in the olfactory bulb at an earlier stage (Braak's stage1) than pathology in the substantia nigra, which is not observed until stage 3. In this report, we investigated the distribution and cell types affected by alpha-synuclein in the olfactory bulb of transgenic mice (2-8 months of age) expressing the human A53T variant of alpha-synuclein. alpha-Synuclein immunostaining progressively affects interneurons and mitral cells. Double labeling studies demonstrate that dopaminergic cells are hardly involved, whereas glutamatergic- and calcium binding protein-positive cells are severely affected. This temporal evolution and the cell types expressing alpha-synuclein are reminiscent of idiopathic Parkinson's disease and support the usefulness of this model to address specific topics in the premotor phase of the disease.

Caspase-11 mediates ischemia-induced astrocyte death: involvement of endoplasmic reticulum stress and C/EBP homologous protein.

Astrocytes are essential cells for maintaining brain integrity. We have recently shown that the transcription factor C/EBP homologous protein (CHOP), associated with endoplasmic reticulum (ER) stress, plays a key role in the astrocyte death induced by ischemia. Meanwhile, mediators of apoptosis downstream of CHOP in the ER stress-dependent pathway remain to be elucidated. Our aim in this work was to determine whether caspase-11, able to activate apoptotic and proinflammatory pathways, is implicated in ER stress-dependent astrocyte death in ischemic conditions. According to our results, caspase-11 is up-regulated in primary astrocyte cultures following either oxygen and glucose deprivation (OGD) or treatment with the ER-stress inducers thapsigargin and tunicamycin. Moreover, these same stimuli increased caspase-11 mRNA levels and luciferase activity driven by a caspase-11 promoter, indicating that caspase-11 is regulated at the transcriptional level. Our data also illustrate the involvement of ER stress-associated CHOP in caspase-11 regulation, insofar as CHOP overexpression by means of an adenoviral vector caused a significant raise in caspase-11. In turn, caspase-11 suppression with siRNA rescued astrocytes from OGD- and ER stress-induced death, supporting the idea that caspase-11 is responsible for the deleterious effects of ischemia on astrocytes. Finally, inhibition of caspase-1 and caspase-3 significantly reduced astrocyte death, which indicates that these proteases act as death effectors of caspase-11. In conclusion, our work contributes to clarifying the pathways leading to astrocyte death in response to ischemia by defining caspase-11 as a key mediator of the ER stress response acting downstream of CHOP.

mTOR/S6 kinase pathway contributes to astrocyte survival during ischemia.

Neurons are highly dependent on astrocyte survival during brain damage. To identify genes involved in astrocyte function during ischemia, we performed mRNA differential display in astrocytes after oxygen and glucose deprivation (OGD). We detected a robust down-regulation of S6 kinase 1 (S6K1) mRNA that was accompanied by a sharp decrease in protein levels and activity. OGD-induced apoptosis was increased by the combined deletion of S6K1 and S6K2 genes, as well as by treatment with rapamycin that inhibits S6K1 activity by acting on the upstream regulator mTOR (mammalian target of rapamycin). Astrocytes lacking S6K1 and S6K2 (S6K1;S6K2-/-) displayed a defect in BAD phosphorylation and in the expression of the anti-apoptotic factors Bcl-2 and Bcl-xL. Furthermore reactive oxygen species were increased while translation recovery was impaired in S6K-deficient astrocytes following OGD. Rescue of either S6K1 or S6K2 expression by adenoviral infection revealed that protective functions were specifically mediated by S6K1, because this isoform selectively promoted resistance to OGD and reduction of ROS levels. Finally, "in vivo" effects of S6K suppression were analyzed in the permanent middle cerebral artery occlusion model of ischemia, in which absence of S6K expression increased mortality and infarct volume. In summary, this article uncovers a protective role for astrocyte S6K1 against brain ischemia, indicating a functional pathway that senses nutrient and oxygen levels and may be beneficial for neuronal survival.

SEPS1 gene is activated during astrocyte ischemia and shows prominent antiapoptotic effects.

Contrarily to neurons, astrocytes can survive short periods of ischemia. We have searched for genes implicated in astrocyte resistance to ischemia using oxygen and glucose deprivation (OGD) as a stroke model. A RNA differential display approach uncovered the OGD induction of selenoprotein-S-encoding gene SEPS1. This endoplasmic reticulum (ER) resident protein is known to promote cell survival regulating the ER stress as well as inflammation. We found that suppression of SEPS1 by small interfering RNA severely increases astrocyte injure caused by OGD, suggesting that selenoprotein S protects astrocytes against ischemia. Our data also support that modulation of ER stress is implicated in this effect.

Mechanical lesion activates newly identified NFATc1 in primary astrocytes: implication of ATP and purinergic receptors.

Ca2+-dependent calcineurin is upregulated in reactive astrocytes in neuroinflammatory models. Therefore, the fact that the nuclear factor of activated T cells (NFAT) is activated in response to calcineurin qualifies this family of transcription factors with immune functions as candidates to mediate astrogliosis. Brain trauma induces a neuroinflammatory state in which ATP is released from astrocytes, stimulating calcium signalling. Our goal here is to characterize NFATc1 and NFATc2 in mouse primary astrocyte cultures, also exploring the implication of NFAT in astrocyte activation by mechanical lesion. Quantitative reverse transcriptase-polymerase chain reaction, Western blot analysis and immunofluorescence microscopy identified NFATc1 in astrocytes, but not NFATc2. Moreover, NFATc1 was expressed in the cytosol of resting astrocytes, whereas activation of the Ca2+-calcineurin pathway by ionomycin translocated NFATc1 to the nucleus, which is a requirement for activation. The implication of astrocytic NFAT in brain trauma was analysed using an in vitro scratch lesion model. Mechanical lesion caused a rapid NFATc1 translocation that progressed throughout the culture as a gradient and was maintained for at least 4 h. We also demonstrate that ATP, released by lesion, is a potent inducer of NFATc1 translocation and activation. Moreover, the use of P2Y receptor modulators showed that such ATP action is mediated by stimulation of several G(q)-protein-coupled P2Y purinergic receptors, among which P2Y(1) and P2Y(6) are included. In conclusion, this work provides evidence that newly identified NFATc1 is translocated in astrocytes in response to lesion following a pathway that involves ATP release and activation of metabotropic purinergic receptors.