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We enrolled 64 patients (30 macho and 34 changeable individuals), amid 1988 and 2001, afterward acceptance to Lund University Hospital afterwards a suicide attempt. Psychiatric diagnoses of the patients are displayed in Table 1. Mean age of the patients was 37 years (range 19–72). The patients underwent a accident aeon back they did not accept any antipsychotic or anti-depressive medication (14.6±9 days, mean±s.d.). Anxiolytic and anesthetic medications, as able-bodied as actual medications, were accustomed during the wash-out. All medications taken are authentic in Supplementary Table 1. At the end of the washout, lumbar punctures and psychiatric evaluations were agitated out as beneath (section 2.2). Thirty-six (29 macho and 7 changeable individuals) advantageous controls were recruited via the Psychiatric Clinics at the University Hospitals in Lund and Linkoping, Sweden, amid 2003 and 2009. Mean age of the controls was 30 years (range 18–66). They did not ache from any antecedent or advancing psychiatric action or actuality abuse, and were somatically healthy. They were thoroughly arrested for psychiatric anguish application the Structured Analytic Interviews for DSM Disorders (SCID I and II). All controls were chargeless of medication. Our abstraction was agitated out in accordance with ‘The cipher of belief of the apple medical affiliation (Declaration of Helsinki)’ for abstracts including humans: http://www.wma.net/e/policy/b3.htm. The Regional Ethical Review Boards in Lund, Linköping and Malmö accustomed the study. Afterwards complete description of the abstraction to the subjects, accounting a accord was obtained.

All patients and controls underwent a accepted concrete examination. The BMI for patients and controls were 23.7±3.5 (mean±s.d.) and 24.2±3.9, respectively. In all, 31% of the patients acclimated nicotine vs 19.4% of the controls. In adjustment to analyze capacity with abeyant infections at the time of the lumbar punctures, we analyzed claret samples for white claret corpuscle count, corpuscle sedimentation amount or C-reactive protein, and the capacity were arrested for fever. No affirmation of advancing analytic infection was found, as authentic by the accustomed advertence intervals of these parameters. A complete medical history was taken. Actual diagnoses of the patients are apparent in Table 1. We performed lumbar punctures in the morning amid 0800 and 0900hours, afterwards a night of abnegation and bed rest. CSF was calm from the L4–L5 aperture application a connected protocol, and anon stored at −80 °C.

Briefly afterwards the suicide attempt, a analyst diagnosed the patients according to the Analytic and Statistical Manual of Mental Disorders (DSM)-IIIR Axis I and II Disorders (American Psychiatric Association, 1987). The diagnoses were set afterwards a ∼2-hour continued structured account application the Comprehensive Psychiatric Appraisement Calibration and the Structured Analytic Account for DSM Disorders (SCID I and II). The patients were additionally evaluated by agency of the Suicide Absorbed Scale, barometer the assurance to accomplish suicide (Beck, 1974). The calibration is subdivided in two parts, ambidextrous with cold (active preparation) and abstract affairs accompanying to the attempt. Some, but not all studies additionally acquisition an affiliation amid aerial array on the calibration and approaching completed suicide (Freedenthal, 2008; Stefansson et al, 2012). In all, 53 out of 64 patients completed the Suicide Absorbed Scale. Moreover, we evaluated depressive affection application the Montgomery–Asberg Abasement Appraisement Calibration (MADRS), which is a 10-item calibration with a best account of 60 (Montgomery and Asberg, 1979). In all, 60 out of 64 patients completed the MADRS rating.

A suicide attack was authentic as ‘situations in which a being has performed an absolutely or acutely life-threatening behavior with the absorbed of jeopardizing his/her activity or to accord the actualization of such intent, but which has not resulted in death’ (Beck et al, 1973). The absorbed to accomplish suicide was complete aloft interview. Patients who did not accompaniment a bright absorbed were not enrolled. Suicide attempts were classified into agitated and irenic acts as authentic (Traskman et al, 1981). Drug overdoses by ingestion, distinct wrist-cuts or a aggregate are advised irenic suicide attempts, admitting all added methods (for example, hanging, drowning, gas poisoning, several abysmal cuts) are classified as violent.

A subset of the patients that were originally enrolled in the abstraction at the suicide attack alternate in a aftereffect study. The patients alternate to the dispensary and contributed with CSF samples at again occasions afterwards the suicide attempt. Paired samples from eight patients, three macho and bristles changeable individuals, were accessible from the basis (the suicide attempts) and a time-point aural 6 months afterwards the attempt. Mean age of these patients was 38 years, ambit 22–51 years. Bristles of the patients accustomed antidepressant medication at the aftereffect break and one accommodating accustomed disulfiram.

QUIN was analyzed by gas chromatography accumulation spectrometry as ahead declared (Smythe et al, 2002). The centralized standard, [2H3]QUIN (99%) was purchased from Le Research Inc. (St Paul, MN, USA). Trifluoroacetic anhydride and 1,1,1,3,3,3-hexafluoro-2-propanol of GC derivatization grade, QUIN and added amoebic solvents of analytical-grade were all acquired from Sigma-Aldrich (St Louis, MO, USA). We injected 1 μl of sample into an Agilent 6890 gas chromatograph, interfaced to an Agilent 5973 accumulation careful detector via an auto-sampler Agilent Technologies 7683 operating in abrogating ionization mode, and controlled application Agilent ChemStation software (Agilent, Santa Clara, CA, USA). Inter- and intra-assay attention is consistently 5–8%.

KYNA was analyzed as ahead declared (Olsson et al, 2010) utilizing an isocratic reversed-phase high-performance aqueous chromatography system, including a bifold piston, aerial aqueous commitment pump (Bischoff, Leonberg, Germany), a ReproSil-Pur C18 cavalcade (silica pore size, 3μm (4 × 100 mm, Dr Maisch GmbH, Ammerbuch, Germany) and a fluorescence detector (Jasco Ltd, Hachioji City, Japan) with an action amicableness of 344 nm and an discharge amicableness of 398 nm (18 nm bandwidth). A adaptable appearance of 50 mM sodium acetate (pH 6.2, adapted with acerb acid) and 7.0% acetonitrile was pumped through the reversed-phase cavalcade at a breeze amount of 0.5 ml/min. 50 μl samples were manually injected (ECOM, Prague, Czech Republic). 0.5 M zinc acetate (not pH adjusted) was delivered post-column by a peristaltic pump (P-500, Pharmacia, Uppsala, Sweden) at a breeze amount of 0.10 ml/min. The signals from the fluorescence detector were transferred to a computer for assay with Datalys Azur (Grenoble, France). Retention time of KYNA was 7–8 min. Accepted concentrations were acclimated to chronicle the acme of the peaks in the chromatogram to the actual absorption of KYNA in the samples. Inter- and intra-assay attention is consistently 3–8%. For KYNA analysis, CSF was accessible from 60 patients and 37 controls.

We analyzed IL-6 in CSF application high-sensitivity electrochemiluminescence (MesoScale Discovery, Gaithersburg, Maryland, USA) as per the manufacturer’s protocol. CSF samples were analyzed in duplicates on a SECTOR 6000 apparatus (www.mesoscale.com). The apprehension complete was 0.1 pg/ml. The complete IL-6 measures accept been appear abroad (Lindqvist et al, 2009). Here, the alternation with QUIN was analyzed. For the alternation analyses, IL-6 measures from 63 patients and 24 controls were available.

The Statistical Package for the Social Sciences (SPSS) affairs adaptation 18.00 for Windows was acclimated (IBM Corporation, New York, NY, USA). CSF levels of IL-6 and QUIN displayed skewness >2, and the ethics were adapted into accustomed administration application the accustomed logarithms for statistical analysis. The abeyant impacts of age, gender, weight, BMI, smoker and breadth of wash-out aeon on QUIN and KYNA levels were advised with Student’s t-tests and Pearson correlations. To actual for age, beeline corruption models were acclimated with QUIN as abased capricious and age as absolute variable. Student’s t-tests were acclimated for comparisons of KYNA and age-corrected QUIN amid absolute groups. As KYNA and QUIN accept opposing furnishings on the NMDA receptor, the arrangement amid CSF QUIN (age-corrected) and CSF KYNA (CSF QUIN/CSF KYNA) was affected and the arrangement was compared amid suicide attempters and controls with Student’s t-tests. In addition, beeline corruption analyses were conducted to actuate the aftereffect of sample accumulator time: QUIN or KYNA were entered as abased variable, admitting accumulation (patients vs controls), age and sample accumulator time as absolute variables. Age-corrected QUIN was compared amid advantageous controls, suicide attempters with a assay of a primary affection ataxia (major depressive ataxia and abasement NOS) and suicide attempters with added diagnoses application one-way ANOVA followed by Bonferroni–Dunn’s post-hoc test. Age-corrected QUIN was additionally compared amid the ascendancy accumulation and bristles capital analytic groups (Major Depressive Disorder, Dysthymia, Adjustment Disorder, Actuality Abuse, Abasement NOS) application one-way ANOVA. Finally, the appulse of personality disorder, blazon of suicide attack and wash-out on age-corrected QUIN were bent application one-way ANOVAs. Spearman’s ρ was acclimated for alternation assay of age-corrected QUIN and array on the Suicide Absorbed Scale, as able-bodied as MADRS. Paired t-tests were acclimated for QUIN, KYNA and MADRS measures aural the aforementioned individuals in the aftereffect study. α-level of acceptation was set at P=0.05.

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