Analysis of health services use and clinical profiles in the year prior to suicide between 2010 and 2018: An opportunity to improve its prevention.

INTRODUCTION: Suicide is the first cause of external death in Spain. International studies show frequent and varied health contacts in the months prior to suicide. There are hardly any studies on that phenomenon in this country. OBJECTIVE: To analyze health care use in the year prior to suicide between 2010 and 2018 in the Basque Country, as well as pharmacological prescriptions and psychiatric diagnoses received. METHODS: Retrospective descriptive study with all suicides registered by the Basque Institute of Legal Medicine (BILM) between 2010 and 2018. The records of the BILM and the Basque Health Service (Osakidetza) were cross-checked. RESULTS: 1526 suicides were analyzed. 74% had health contacts in the previous year. The use was higher in women (p<0.05) and in older ages (p<0001). Primary care was the most used specialty (58.8% the previous year and 7.1% the previous week), followed by Hospital Emergencies (50.3% and 10.2%) and Outpatient Medical Specialties (49% and 11.6%), especially Radiology. Outpatient psychiatry only contacted 29.6% that year, although it had the highest average number of visits (15.1 SD22.6). The most frequent diagnostic category among psychiatric patients was F30-39 (26.7%), with differences between sexes and ages. 49.7% received psychotropic drugs. CONCLUSIONS: The results are aligned with international evidence, which they also extend, and reinforce the need to extend prevention beyond psychiatric services. It seems advisable to increase proactivity in the search for risk by sensitizing and training different professional profiles, but also to work from non-health settings to improve assistance to highly vulnerable profiles (young men) with low health links.

Alcohol Triggers the Accumulation of Oxidatively Damaged Proteins in Neuronal Cells and Tissues.

Alcohol is toxic to neurons and can trigger alcohol-related brain damage, neuronal loss, and cognitive decline. Neuronal cells may be vulnerable to alcohol toxicity and damage from oxidative stress after differentiation. To consider this further, the toxicity of alcohol to undifferentiated SH-SY5Y cells was compared with that of cells that had been acutely differentiated. Cells were exposed to alcohol over a concentration range of 0-200 mM for up to 24 h and alcohol effects on cell viability were evaluated via MTT and LDH assays. Effects on mitochondrial morphology were examined via transmission electron microscopy, and mitochondrial functionality was examined using measurements of ATP and the production of reactive oxygen species (ROS). Alcohol reduced cell viability and depleted ATP levels in a concentration- and exposure duration-dependent manner, with undifferentiated cells more vulnerable to toxicity. Alcohol exposure resulted in neurite retraction, altered mitochondrial morphology, and increased the levels of ROS in proportion to alcohol concentration; these peaked after 3 and 6 h exposures and were significantly higher in differentiated cells. Protein carbonyl content (PCC) lagged behind ROS production and peaked after 12 and 24 h, increasing in proportion to alcohol concentration, with higher levels in differentiated cells. Carbonylated proteins were characterised by their denatured molecular weights and overlapped with those from adult post-mortem brain tissue, with levels of PCC higher in alcoholic subjects than matched controls. Hence, alcohol can potentially trigger cell and tissue damage from oxidative stress and the accumulation of oxidatively damaged proteins.

Permissive epigenetic regulatory mechanisms at the histone level are enhanced in postmortem dorsolateral prefrontal cortex of individuals with schizophrenia.

BACKGROUND: Susceptibility to schizophrenia is determined by interactions between genes and environment, possibly via epigenetic mechanisms. Schizophrenia has been associated with a restrictive epigenome, and histone deacetylase (HDAC) inhibitors have been postulated as coadjuvant agents to potentiate the efficacy of current antipsychotic drugs. We aimed to evaluate global histone posttranslational modifications (HPTMs) and HDAC expression and activity in the dorsolateral prefrontal cortex (DLPFC) of individuals with schizophrenia. METHODS: We used postmortem DLPFC samples of individuals with schizophrenia and controls matched for sex, age, and postmortem interval. Schizophrenia samples were classified into antipsychotic-treated or antipsychotic-free subgroups according to blood toxicology. Expression of HPTMs and HDAC was quantified by Western blot. HDAC activity was measured with a fluorometric assay. RESULTS: H3K9ac, H3K27ac, and H3K4me3 were globally enhanced in the DLPFC of individuals with schizophrenia (+24%-42%, p < 0.05). HDAC activity (-17%, p < 0.01) and HDAC4 protein expression (-20%, p < 0.05) were downregulated in individuals with schizophrenia. Analyses of antipsychotic-free and antipsychotic-treated subgroups revealed enhanced H3K4me3 and H3K27ac (+24%-49%, p < 0.05) and reduced HDAC activity in the antipsychotic-treated, but not in the antipsychotic-free subgroup. LIMITATIONS: Special care was taken to control the effect of confounding factors: age, sex, postmortem interval, and storage time. However, replication studies in bigger cohorts might strengthen the association between permissive HPTMs and schizophrenia. CONCLUSION: We found global HPTM alterations consistent with an aberrantly permissive epigenome in schizophrenia. Further studies to elucidate the significance of enhanced permissive HPTMs in schizophrenia and its association with the mechanism of action of antipsychotic drugs are encouraged.

Effect of antipsychotic drugs on group II metabotropic glutamate receptor expression and epigenetic control in postmortem brains of schizophrenia subjects.

Antipsychotic-induced low availability of group II metabotropic glutamate receptors (including mGlu2R and mGlu3R) in brains of schizophrenia patients may explain the limited efficacy of mGlu2/3R ligands in clinical trials. Studies evaluating mGlu2/3R levels in well-designed, large postmortem brain cohorts are needed to address this issue. Postmortem samples from the dorsolateral prefrontal cortex of 96 schizophrenia subjects and matched controls were collected. Toxicological analyses identified cases who were (AP+) or were not (AP-) receiving antipsychotic treatment near the time of death. Protein and mRNA levels of mGlu2R and mGlu3R, as well as GRM2 and GRM3 promoter-attached histone posttranslational modifications, were quantified. Experimental animal models were used to compare with data obtained in human tissues. Compared to matched controls, schizophrenia cortical samples had lower mGlu2R protein amounts, regardless of antipsychotic medication. Downregulation of mGlu3R was observed in AP- schizophrenia subjects only. Greater predicted occupancy values of dopamine D2 and serotonin 5HT2A receptors correlated with higher density of mGlu3R, but not mGlu2R. Clozapine treatment and maternal immune activation in rodents mimicked the mGlu2R, but not mGlu3R regulation observed in schizophrenia brains. mGlu2R and mGlu3R mRNA levels, and the epigenetic control mechanisms did not parallel the alterations at the protein level, and in some groups correlated inversely. Insufficient cortical availability of mGlu2R and mGlu3R may be associated with schizophrenia. Antipsychotic treatment may normalize mGlu3R, but not mGlu2R protein levels. A model in which epigenetic feedback mechanisms controlling mGlu3R expression are activated to counterbalance mGluR loss of function is described.

Toxicology Screening in Sports-Related Sudden Cardiac Death: A Multinational Observational Study.

BACKGROUND: Knowledge of toxicological findings among sports-related sudden cardiac death (SrSCD) is scarce. OBJECTIVES: This study aimed to describe postmortem toxicology findings in a multinational cohort of young SrSCD. METHODS: Patients with sudden cardiac death (SCD) aged 12 to 49 years with a complete post mortem were included from Denmark, Spain, and Australia. Postmortem findings were compared between SrSCD and non-SrSCD, and toxicology findings in SrSCD were assessed. RESULTS: We included 3,189 SCD, of which 219 (7%) were sports-related. SrSCD patients were younger (36 years vs 41 years; P < 0.001) and of male predominance (96% vs 75%; P < 0.001), and their death was more often caused by structural cardiac disease (68% vs 61%; P = 0.038). Positive toxicology screenings were significantly less likely among SrSCD than non-SrSCD (12% vs 43%; P < 0.001), corresponding to 82% lower odds of a positive toxicology screening in SrSCD. Patient characteristics were similar between SrSCDs with positive and negative toxicology screenings, but deaths were more often unexplained (59% vs 34%). Nonopioid analgesics were the most common finding (3%), and SCD-associated drugs were detected in 6% of SrSCD. SUD was more prevalent among the SrSCD with positive toxicology (59% vs 34%). CONCLUSIONS: Sports-related SCD mainly occurred in younger men with structural heart disease. They had a significantly lower prevalence of a positive toxicology screening compared with non-SrSCD, and detection of SCD-associated drugs was rare.

Ethanol and illicit drugs acute use and abuse as risk factors for suicide: A case-control study based on forensic autopsies in the Basque Country, Spain.

INTRODUCTION: Abuse/dependence and acute use of ethanol and illicit drug are considered risk factors for suicide. The risk is also influenced by demographic conditions and/or psychiatric comorbidity. The aim of the study was to test the association between presence of ethanol, illicit substances and prescribed drugs in suicide decedents and controls. MATERIALS AND METHODS: Case-control study of autopsies performed in the Biscay Forensic Pathology Service, Basque Country, Spain from 01/01/2010 to 30/06/2021 in subjects between 15 and 55 years old. Suicide deaths (n=481) with completed autopsy were evaluated. Concurrent natural deaths were chosen as controls (n=330). The risk for suicide according to demographic, toxicological and psychiatric variables was analyzed using logistic regression. RESULTS: Ethanol was present in 21% and illicit drugs, mainly cannabis, cocaine and amphetamine, in 27% of suicide deaths. Illicit drugs were more frequent among males. In 63% of suicide cases, prescribed psychotropic drugs were detected. In a multivariate analysis, the main risk factors for suicide were psychiatric diagnosis of illicit drug abuse/dependence (OR=5.56, 95% CI 2.74-11.30) or another mental disease as mood or psychotic disorders (OR=13.05, 95% CI 8.79-19.37). Acute presence of ethanol (OR=4.22, 95% CI 2.52-7.08), recent use of cocaine (OR=2.52, 95% CI 1.05-6.07) and age <35 years (OR=2.50, 95% CI 1.62-3.87) were also associated with suicide deaths. CONCLUSIONS: The presence of drugs of abuse in suicide deaths of people ≤55 years old is high. Recent use of ethanol and cocaine is significantly associated with an increased suicide risk. Specific prevention strategies against exposition to substances of abuse should be promoted, especially in psychiatric patients.

Differential brain ADRA2A and ADRA2C gene expression and epigenetic regulation in schizophrenia. Effect of antipsychotic drug treatment.

Postsynaptic α2A-adrenoceptor density is enhanced in the dorsolateral prefrontal cortex (DLPFC) of antipsychotic-treated schizophrenia subjects. This alteration might be due to transcriptional activation, and could be regulated by epigenetic mechanisms such as histone posttranslational modifications (PTMs). The aim of this study was to evaluate ADRA2A and ADRA2C gene expression (codifying for α2-adrenoceptor subtypes), and permissive and repressive histone PTMs at gene promoter regions in the DLPFC of subjects with schizophrenia and matched controls (n = 24 pairs). We studied the effect of antipsychotic (AP) treatment in AP-free (n = 12) and AP-treated (n = 12) subgroups of schizophrenia subjects and in rats acutely and chronically treated with typical and atypical antipsychotics. ADRA2A mRNA expression was selectively upregulated in AP-treated schizophrenia subjects (+93%) whereas ADRA2C mRNA expression was upregulated in all schizophrenia subjects (+53%) regardless of antipsychotic treatment. Acute and chronic clozapine treatment in rats did not alter brain cortex Adra2a mRNA expression but increased Adra2c mRNA expression. Both ADRA2A and ADRA2C promoter regions showed epigenetic modification by histone methylation and acetylation in human DLPFC. The upregulation of ADRA2A expression in AP-treated schizophrenia subjects might be related to observed bivalent chromatin at ADRA2A promoter region in schizophrenia (depicted by increased permissive H3K4me3 and repressive H3K27me3) and could be triggered by the enhanced H4K16ac at ADRA2A promoter. In conclusion, epigenetic predisposition differentially modulated ADRA2A and ADRA2C mRNA expression in DLPFC of schizophrenia subjects.

Sports-related sudden cardiac death in Spain. A multicenter, population-based, forensic study of 288 cases.

INTRODUCTION AND OBJECTIVES: To determine the incidence of sports-related sudden cardiac death in Spain, and to identify the clinical-pathological characteristics, substance abuse, and sports activity associated with this entity. METHODS: Retrospective, population-based, multicenter study of forensic autopsies conducted in 25 provinces during an 8-year period (2010-2017). RESULTS: We investigated 288 cases (98.6% occurred in men with a mean age of 43.8±14.4 years). The incidence in the general population was 0.38 cases out of 100 000 inhabitants per year (0.82 among regular athletes), and most cases (54%) occurred in persons aged between 35 and 54 years. The most frequent sports (96% recreational) were cycling (28%), football (18%), and jogging (17%). Death was of cardiovascular origin in 99%. The main causes were ischemic heart disease (63%), cardiomyopathies (21%), and sudden arrhythmic death syndrome (6%). In young people, cardiomyopathies (38%) and ischemic heart disease (30%), present after the age of 20 years, were the most prevalent. The disease was diagnosed during life in 23 cases, relevant clinical antecedents were observed in 30 cases, and cardiovascular risk factors, mainly obesity, in 95 cases. Toxicological analysis detected cardiotoxic substances in 7%, highlighting the association between cannabis and acute ischemic heart disease. CONCLUSIONS: Sports-related sudden cardiac death in Spain has a very low incidence and affects middle-aged men practicing recreational sports, mainly cycling, football, and jogging. This entity is of cardiovascular origin with early onset of ischemic heart disease. Clinical data and substance abuse should be taken into account to develop preventive strategies.

Ribosomal Protein S6 Hypofunction in Postmortem Human Brain Links mTORC1-Dependent Signaling and Schizophrenia.

The mechanistic target of rapamycin (also known as mammalian target of rapamycin) (mTOR)-dependent signaling pathway plays an important role in protein synthesis, cell growth, and proliferation, and has been linked to the development of the central nervous system. Recent studies suggest that mTOR signaling pathway dysfunction could be involved in the etiopathogenesis of schizophrenia. The main goal of this study was to evaluate the status of mTOR signaling pathway in postmortem prefrontal cortex (PFC) samples of subjects with schizophrenia. For this purpose, we quantified the protein expression and phosphorylation status of the mTOR downstream effector ribosomal protein S6 as well as other pathway interactors such as Akt and GSK3ß. Furthermore, we quantified the status of these proteins in the brain cortex of rats chronically treated with the antipsychotics haloperidol, clozapine, or risperidone. We found a striking decrease in the expression of total S6 and in its active phosphorylated form phospho-S6 (Ser235/236) in the brain of subjects with schizophrenia compared to matched controls. The chronic treatment with the antipsychotics haloperidol and clozapine affected both the expression of GSK3ß and the activation of Akt [phospho-Akt (Ser473)] in rat brain cortex, while no changes were observed in S6 and phospho-S6 (Ser235/236) protein expression with any antipsychotic treatment. These findings provide further evidence for the involvement of the mTOR-dependent signaling pathway in schizophrenia and suggest that a hypofunctional S6 may have a role in the etiopathogenesis of this disorder.

Serotonin 5-HT2A receptor expression and functionality in postmortem frontal cortex of subjects with schizophrenia: Selective biased agonism via Gαi1-proteins.

Serotonin 5-HT2A receptors (5-HT2ARs) have been implicated in schizophrenia. However, postmortem studies on 5-HT2ARs expression and functionality in schizophrenia are scarce. The 5-HT2AR mRNA and immunoreactive protein expression were evaluated in postmortem tissue from dorsolateral prefrontal cortex (DLPFC) of antipsychotic-free (n = 18) and antipsychotic-treated (n = 9) subjects with schizophrenia, and matched controls (n = 27). Functional coupling of 5-HT2AR to G-proteins was tested by measuring the activation induced by the agonist (±)-2,5-dimethoxy-4-iodoamphetamine hydrochloride ((±)DOI) in antibody-capture [35S]GTPγS scintillation proximity assays (SPA). In antipsychotic-free schizophrenia subjects, 5-HT2AR mRNA expression and protein immunoreactivity in total homogenates was similar to controls. In contrast, in antipsychotic-treated schizophrenia subjects, lower mRNA expression (60±9% vs controls) and a trend to reduced protein immunoreactivity (86±5% vs antipsychotic-free subjects) just in membrane-enriched fractions was observed. [35S]GTPγS SPA revealed a significant ~6% higher stimulation of Gαi1-protein by (±)DOI in schizophrenia, whereas activation of the canonical Gαq/11-protein pathway by (±)DOI remained unchanged. Expression of Gαi1- and Gαq/11-proteins did not differ between groups. Accordingly, in rats chronically treated with clozapine, but not with haloperidol, a 30-40% reduction was observed in 5-HT2AR mRNA expression, 5-HT2AR protein immunoreactivity and [3H]ketanserin binding in brain cortical membranes. Overall, the data suggest a supersensitive 5-HT2AR signaling through inhibitory Gαi1-proteins in schizophrenia. Together with previous results, a dysfunctional pro-hallucinogenic agonist-sensitive 5-HT2AR conformation in postmortem DLPFC of subjects with schizophrenia is proposed. Atypical antipsychotic treatment would contribute to counterbalance this 5-HT2AR supersensitivity by reducing receptor expression.

Sports-related sudden cardiac death due to myocardial diseases on a population from 1-35 years: a multicentre forensic study in Spain.

This is a multicentre forensic study that identifies all sports-related sudden deaths (SRSDs) in young people, due to myocardial diseases (MDs) that occurred in a large area of Spain. The aim of the study is to assess the epidemiology, causes of death, and sport activities associated with these fatalities. This is a retrospective study based on forensic autopsies performed in the provinces of Biscay, Seville, Valencia and in the jurisdiction covered by the National Institute of Toxicology and Forensic Sciences in Madrid (Spain). The retrospective study encompasses from 2010 to 2017. All sudden cardiac deaths (SCDs) in persons 1-35 years old were selected. The total number of SCDs were divided into death occurred during exercise (SRSD) and death during rest, sleep or normal activities (non-SRSD). Each of these two groups was subdivided according to the cause of death into MD (primary cardiomyopathies and myocarditis) and non-MD. Clinic-pathological, toxicological and genetic characteristics of SRSD due to MD were analysed. Over the 8-year study period, we identified 645 cases of SCD in the young: 75 SRSD (11.6%) and 570 non-SRSD (88.4%). MD was diagnosed in 33 (44.0%) of the SRSD and in 112 (19.6%) of the non-SRSD cases. All cases of SRSD due to MD were males (mean age (24.0 ± 7.6) years) practicing recreational sports (85%). SRSDs were more frequent in arrhythmogenic cardiomyopathy (ACM) (37%) and hypertrophic cardiomyopathy (HCM) (24%), followed by myocarditis (15%) and idiopathic left ventricular hypertrophy (ILVH) (9%). Only in five cases of SRSD the MD responsible of death (HCM) had been diagnosed in life. Cardiovascular symptoms related to the disease were present in other seven patients (six of them with ACM). Postmortem genetic studies were performed in 15/28 (54%) primary cardiomyopathies with positive results in 12 (80%) cases. The most frequent sports disciplines were football (49%) followed by gymnastics (15%) and running (12%). In Spain, SRSD in young people due to MDs occurs in males who perform a recreational activity. Compared with control group we observed a strong association between MDs and exertion. One in three SRSDs are due to cardiomyopathy, especially ACM, which reinforces the need for preparticipation screening to detect these pathologies in recreational sport athletes. Further studies are warranted to understand the causes and circumstances of sudden death to facilitate the development of preventive strategies.

Endocannabinoid system imbalance in the postmortem prefrontal cortex of subjects with schizophrenia.

BACKGROUND: The endocannabinoid system - comprising cannabinoid receptors, endocannabinoid ligands and their synthesis and inactivation enzymes - has been widely implicated in the pathophysiology of schizophrenia. However, little is known regarding the status of the different elements of the endocannabinoid system in the brain of schizophrenic patients. We have previously reported altered endocannabinoid levels in the postmortem brain of subjects with schizophrenia compared with matched controls. AIMS: Our aim was to further examine the status of the main elements of the endocannabinoid system in the postmortem prefrontal cortex of the same cohort of subjects. METHODS: Gene expression and function of the cannabinoid receptor type-1 (CB1) and the endocannabinoid degrading enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) have been assessed. RESULTS: A significant decrease in CB1 mRNA levels in schizophrenia was found, without alteration of FAAH or MAGL mRNA expression. Moreover, positive correlations among mRNA expressions of the three genes studied were found in the prefrontal cortex of controls but not in schizophrenic subjects. No alteration was found in CB1 receptor mediated functional coupling to G-proteins, but a significant increase of FAAH activity was found in schizophrenic subjects compared with controls. 2-arachidonoylglycerol levels and MAGL activity were found to positively correlate in controls but not in schizophrenic subjects. CONCLUSIONS: The present findings reveal an imbalance in the expression and function of different elements of the endocannabinoid system in schizophrenia. This outcome highlights the relevance of the endocannabinoid system in the pathophysiology of schizophrenia and emphasises its elements as potential targets in the search for new therapeutic strategies.

Sudden cardiac death associated to substances of abuse and psychotropic drugs consumed by young people: A population study based on forensic autopsies.

BACKGROUND: Toxic substances are one of the main risk factors for sudden cardiac death (SCD) in young people. However, there is limited information about this matter based on clinical research. The aim of this study was to analyze the use of substances of abuse (legal and illicit) and prescribed psychotropic drugs in young people who died by SCD. METHODS: A population-based study performed in 15-35-year-olds who died by SCD in Biscay (Basque-Country) between 1991 and 2016. Cases were analyzed prospectively by a complete autopsy, toxicological and histopathological studies. A case was considered positive for exposure to cardiotoxic substances if smoking status was diagnosed or if toxicological analysis detected any drug associated with increased risk of SCD. RESULTS: There were 204 SCD; 98 (48%) were exposed to a cardiotoxic substance, including smoking status (n = 72) and/or positive toxicology (n = 58). Illicit drugs (n = 29, mainly cannabis and cocaine), ethanol (n = 25), and prescribed psychotropic drugs (n = 11) were detected. Positive cases were more frequent in males than in females (54% vs. 19%). They were also more common in subjects who died by acute (86%) and chronic (71%) ischemic heart disease than in myocardial diseases (33%) and sudden arrhythmic death syndrome (36%). All positive cases of illicit drugs were males. Smoking status was very high in deaths due to acute ischemic heart disease. CONCLUSIONS: The proportion of users of substances of abuse was unexpectedly high, even more prevalent than other cardiovascular risk factors. Toxic substances could play an important role as triggers of SCD in young people.

Assessing the level of credibility of allegations of physical torture.

BACKGROUND: No means exists to classify the overall credibility of physical torture allegations, which relies on qualitative forensic medical criteria. OBJECTIVES: To develop procedures for processing physical data from torture allegations in order to achieve robust credibility assessments. To assess inter-observer variation in semi-quantitative evaluations. To analyse the importance of physical aspects for the overall assessment of allegations of torture. To assess the correlation between a quantitative description of physical aspects of torture and the appraisal of credibility based on the semi-quantitative model. METHODS: We developed a semi-quantitative model to assess credibility of physical torture allegations and a model to describe allegations quantitatively. Information about physical aspects of ill-treatment in 202 extensive reports made by psychologists (part of a Basque Country Governmental project) was appraised individually and jointly by the two authors in the semi-quantitative model. Inter-observer variation was appraised. The assessed credibility based on physical details was compared with the psychologists' overall assessments and with the quantitative scoring of physical data. FINDINGS: The inter-observer agreement was substantial with kappa values of 0.64-0.77. Peer-reviewing revealed an error; using the defined criteria in the models facilitated achievement of common opinions. Comparison of credibility assessments found by different means suggests that physical data may be given a low priority in the overall assessment of torture allegations. There was a strong correlation (r=0,86, p<0.001) between the semi-quantitative credibility assessment and the quantitative scoring of physical aspects of torture, reinforcing the validity of both means. CONCLUSION: Peer-reviewing of physical aspects of torture based on a qualitative /semi-quantitative /quantitative multi-step analysis leads to robust assessments of credibility. A procedure is needed on how to merge the credibility assessments of the psychological and the physical data into an overall conclusion. We suggest that the highest level of the two evaluations should constitute the overall credibility assessment. LIMITATIONS: The examinations were done years after the detentions, they were not focused on physical details and the data we used was retrieved retrospectively.

The Loss of α- and ß-Tubulin Proteins Are a Pathological Hallmark of Chronic Alcohol Consumption and Natural Brain Ageing.

Repetitive excessive alcohol intoxication leads to neuronal damage and brain shrinkage. We examined cytoskeletal protein expression in human post-mortem tissue from Brodmann's area 9 of the prefrontal cortex (PFC). Brain samples from 44 individuals were divided into equal groups of 11 control, 11 alcoholic, 11 non-alcoholic suicides, and 11 suicide alcoholics matched for age, sex, and post-mortem delay. Tissue from alcoholic cohorts displayed significantly reduced expression of α- and ß-tubulins, and increased levels of acetylated α-tubulin. Protein levels of histone deacetylase-6 (HDAC6), and the microtubule-associated proteins MAP-2 and MAP-tau were reduced in alcoholic cohorts, although for MAPs this was not significant. Tubulin gene expressions increased in alcoholic cohorts but not significantly. Brains from rats administered alcohol for 4 weeks also displayed significantly reduced tubulin protein levels and increased α-tubulin acetylation. PFC tissue from control subjects had reduced tubulin protein expression that was most notable from the sixth to the eighth decade of life. Collectively, loss of neuronal tubulin proteins are a hallmark of both chronic alcohol consumption and natural brain ageing. The reduction of cytosolic tubulin proteins could contribute to the brain volumetric losses reported for alcoholic patients and the elderly.

Detainees' perception of the doctors and the medical institution in Spanish police stations: An impediment in the fight against torture and ill-treatment.

BACKGROUND: As part of a program by the Basque Government (Spain) and the University of the Basque Country, persons who have alleged exposure to torture and ill-treatment have been examined by psychologists and psychiatrists according to the Istanbul Protocol (IP). Medical examinations of detainees with the aim, inter alia, to document abuses is fundamental for torture prevention. The IP prescribes how this should be done to ensure data collection and prevent reprisals for having reported ill-treatment to the doctor. OBJECTIVES: The objectives were to assess detainees' perception of the medical examinations which they underwent at different types of Spanish police institution and to compare practice between institutions; and, to compare information from the period 1969-1991 with that from 1992-2014. METHODS: All information about medical examinations of incommunicado detainees from 202 extensive IP reports was analysed according to a 19-item tool developed for the purpose dealing with the doctors' professional performance, the confidence of the detainees in the doctors, reprisals and procedural safeguards (Annex 1). All information was classified as acceptable, unacceptable /insufficient or totally unacceptable. FINDINGS: Very often the detainees perceived the doctors' professional performance as insufficient or totally unacceptable and the doctors did not instil their confidence. Threats of police reprisals and failure to observe procedural safeguards were often reported. There were no differences in the comparison between institutions. When comparing the two study periods an improvement was found over time. However, 64% of the scores came out as totally unacceptable or insufficient for the most recent period indicating that the medical institution in police stations where the detentions had taken place failed to constitute a preventive means against torture and ill-treatment. LIMITATIONS: Only 57% of the 202 reports assessed contained relevant information; data was retrieved retrospectively; the interviews were done years after detention without a particular focus on medical examinations.

Incommunicado detention and torture in Spain, Part II: Enhanced credibility assessment based on the Istanbul Protocol.

BACKGROUND: The Istanbul Protocol (IP) is the key instrument in the documentation of allegations of torture. However, few scientific studies have evaluated its effectiveness as a tool to assess credibility of allegations of ill-treatment or torture. OBJECTIVE: Present data on the credibility of allegations of torture in a sample of 45 Basque people held in short-term incommunicado detention between 1980 and 2012, using a modified version of the Standard Evaluation Form for Credibility Assessment (SEC), a new tool to assess credibility based on the IP. METHOD: Each case was evaluated by two psychiatrists, a psychologist and a physician through a layered system of simultaneous, independent assessments, blind audits and peer-review processes. Clinical interviews following the IP were contrasted with psychometric tests and external documentary evidence by independent experts. All available data were structured using the SEC and cases were accordingly classified as having Maximum consistency, Highly Consistent, Consistent or Inconsistent. FINDINGS: According to the SEC, 53% of allegations of torture were considered to have Maximum Consistency, 31% Highly consistent, 15% Consistent and 0% Inconsistent. The items that most contributed to the overall credibility assessment came from the psychological evaluation, including the description of alleged torture, emotional reactions, objective functional changes, changes in identity and worldviews and clinical diagnosis. There was little contribution from previous medical reports. INTERPRETATION: When applied competently, the IP is an essential tool in the documentation of torture. Our study shows: (a) evidence that allegations of ill-treatment and torture in the Basque Country are consistent and credible, being ascertained beyond reasonable doubt and aside from any political debate; (b) the wider use of the IP as a tool to assess credibility of allegations of ill-treatment and torture; and, (c) the usefulness of the SEC as a tool. The SEC can help as a tool for documenting torture in contexts where there are political differences and figures are distorted as a result of polarized political debates, and where legal documentation is needed for judicial purposes. Forensic science can help by providing an objective assessment of the credibility of allegations.

Incommunicado detention and torture in Spain, Part III: 'Five days is enough': the concept of torturing environments.

BACKGROUND: Torture is changing in western societies, evolving from pain-producing torture to more subtle mixed psychological methods that are harder to detect. Despite this, there is not an adequate understanding of the complexities of contemporary psychological techniques used in coercive interrogation and torture. METHODS: The interrogation and torture techniques used on 45 detainees held in short-term incommunicado detention in Spain during the period 1980-2012 were analyzed. The list of torture categories set out in the Istanbul Protocol (IP) were assessed quantitatively. Software-aided qualitative analysis of the testimonies was conducted, using both inferential and deductive approaches to deduce a classification of torture techniques from the point of view of the survivor. FINDINGS: The most frequent methods according to the IP categories used against detainees were isolation and manipulation of environment (100%), humiliation (93%), psychological techniques to break down the individual (91%), threats (89%) and forced positions and physical exercises until extenuation (80%). Additionally, with a frequency of between 51 and 70%, mild but constant blows, being forced to witness the torture of others, hooding (mainly dry asphyxia) and unacceptable undue conditions of detention were also frequent. Sexual torture was also widespread with sexual violence (42%), forced nudity (38%) and rape (7%). Qualitative analysis showed that most detainees were submitted to coercive interrogation using a wide array of deceptive techniques. This is often a central part of the torturing process, frequently used in conjunction with many other methods. It was found that giving false or misleading information or making false accusations was most frequently used, followed by maximization of responsibility or facts and giving false information regarding relatives or friends. Different patterns of harsh interrogation, ill-treatment and torture are described that appear to have been tailored to the profile of Basque detainees. INTERPRETATION: The study shows the need to improve the conceptualization of psychological torture suggested by the IP. Key to this view is the idea that we must not concern ourselves with 'torture methods' but with Torturing Environments. The concept of Torturing Environments is defined and proposed as a focus for future study.

Incommunicado detention and torture in Spain, Part IV: Psychological and psychiatric consequences of ill-treatment and torture: trauma and human worldviews.

BACKGROUND: Most literature on psychological consequences of torture is related to prolonged detention. Psychological consequences of intensive physical and psychological torture in brief detention have not been investigated. The aim of this study is to analyse the psychological impact of torture in short-term incommunicado detention. METHOD: A sample of 45 Basque people who had allegedly been subjected to ill-treatment or torture whilst held in incommunicado detention between two and 11 days in Spain in the period 1980-2012 was analysed. The period between detention and evaluation ranged between two and 12 years. Each case was evaluated by several psychiatrists and psychologists. Clinical interviews which followed the Istanbul Protocol were assessed, as were psychometric tests (Post-traumatic Checklist-Civilian version (PCL-C), Beck Depression Inventory (BDI) and Vital Impact Assessment Questionnaire (VIVO)) and external documentary evidence. A cumulative prevalence of psychiatric diagnosis (ICD-10) from the period of detention was established. FINDINGS: Post-traumatic stress disorder (PTSD) was the most frequent diagnosis (53%). Enduring personality change after a catastrophic experience was detected in 11% of subjects. Other diagnoses were depressive disorders (16%) and anxiety disorders (9%). Psychometric evaluation at the time of the study showed symptoms of PTSD in 52% of the subjects (with a tendency for these symptoms to diminish over time) and depressive symptoms in 56%. The VIVO questionnaire discerned two subgroups of survivors: "affected" survivors (36%); and "resilient" survivors (64%). INTERPRETATION: The data demonstrated two important issues: the undervalued damaging effect of intensive torture in short-term detention and the long lasting psychological damage of the same over time.

Up-regulated 14-3-3ß and 14-3-3ζ proteins in prefrontal cortex of subjects with schizophrenia: effect of psychotropic treatment.

14-3-3 is a family of conserved regulatory proteins that bind to a multitude of functionally diverse signalling proteins. Various genetic studies and gene expression and proteomic analyses have involved 14-3-3 proteins in schizophrenia (SZ). On the other hand, studies about the status of these proteins in major depressive disorder (MD) are still missing. Immunoreactivity values of cytosolic 14-3-3ß and 14-3-3ζ proteins were evaluated by Western blot in prefrontal cortex (PFC) of subjects with schizophrenia (SZ; n=22), subjects with major depressive disorder (MD; n=21) and age-, gender- and postmortem delay-matched control subjects (n=52). The modulation of 14-3-3ß and 14-3-3ζ proteins by psychotropic medication was also assessed. The analysis of both proteins in SZ subjects with respect to matched control subjects showed increased 14-3-3ß (Δ=33±10%, p<0.05) and 14-3-3ζ (Δ=29±6%, p<0.05) immunoreactivity in antipsychotic-free but not in antipsychotic-treated SZ subjects. Immunoreactivity values of 14-3-3ß and 14-3-3ζ were not altered in MD subjects. These results show the specific up-regulation of 14-3-3ß and 14-3-3ζ proteins in PFC of SZ subjects and suggest a possible down-regulation of both proteins by antipsychotic treatment.