Marina Mihaljevic

Lymphocyte levels of redox-sensitive transcription factors and antioxidative enzymes as indicators of pro-oxidative state in depressive patients.

Background: Oxidative stress is reliably observed in major depressive disorder (MDD). However, molecular data on the principal cellular redox-sensitive transcriptional factors and the levels of their downstream-regulated antioxidant enzymes in MDD are scarce. Methods: In the peripheral blood mononuclear cells (PBMC) of subjects with a current episode of MDD (n = 30) and healthy controls (n = 35), we investigated alterations in the levels of redox-sensing nuclear factor (erythroid-derived 2)-like 2 (Nrf2) protein, its inhibitor Keap1, and nuclear factor-κB (NF-κB), along with their cognate downstream effectors, the antioxidant enzymes (AOEs): manganese and copper zinc superoxide dismutase (MnSOD and CuZnSOD, respectively), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GLR). Results: MDD subjects exhibited higher levels of Nrf2 and its regulator Keap1, as well as NF-κB in the cytoplasm of PBMC compared to controls. This state was further reflected by increased levels of MnSOD, CuZnSOD and CAT proteins and by the lack of correlation between MnSOD and CAT, which could indicate impaired oxidative detoxification capacity in MDD patients. Moreover, increased levels of MnSOD, CuZnSOD and CAT in MDD patients positively correlated with levels of Nrf2, while increased levels of SODs were also positively related to NF-κB. There were no differences regarding the levels of GPx and GLR proteins, but the ratio of GLR/GPx was reduced, suggesting diminished capacity of GPx in antioxidative defence in PBMC of MDD subjects. Conclusion: These data provide evidence that MDD is characterized by up-regulation of redox-sensitive transcriptional factors (Nrf2 and NF-κB) and AOEs (MnSOD, CuZnSOD and CAT), indicating pro-oxidative state in the PBMC of MDD patients.

Stigmatization of ‘psychiatric label’ by medical and non-medical students

Backround: Stigmatization of psychiatric patients is present both in the general population and among healthcare professionals. Aim: To determine the attitudes and behaviour of medical students towards a person who goes to a psychiatrist, before and after psychiatric rotation, and to compare those attitudes between medical and non-medical students. Methods: The study included 525 medical students (second and sixth year of studies) and 154 students of law. The study instrument was a three-part self-reported questionnaire (socio-demographic data, Rosenberg Self-Esteem Scale and a vignette depicting a young, mentally healthy person). The experimental intervention consisted of ascribing a ‘psychiatric label’ to only one set of vignettes. All the vignettes (with or without the ‘psychiatric label’) were followed by 14 statements addressing the acceptance of a person described by vignette, as judged by social distance (four-point Likert scale). Results: Higher tendency to stigmatize was found in medical students in the final year, after psychiatric rotation (Z U = −3.12, p = .002), particularly in a closer relationship (Z U = −2.67, p = .007) between a student and a hypothetical person who goes to a psychiatrist. The non-medical students had a similar tendency to stigmatize as medical students before psychiatric rotation (Z U = −0.03, p = .975). Neither gender, nor the size of student’s place of origin or average academic mark was associated with the tendency to stigmatize in our sample. However, student’s elf-esteem was lower in those with a tendency to stigmatize more in a distant relationship (ρ = −0.157, p = .005). Conclusions: Psychiatric education can either reinforce stigmatization or reduce it. Therefore, detailed analyses of educational domains that reinforce stigma will be the starting point for anti-stigma action.

The emerging role of the FKBP5 gene polymorphisms in vulnerability-stress model of schizophrenia: further evidence from a Serbian population

Increased reactivity to stress is observed in patients with schizophrenia spectrum disorders and their healthy siblings in comparison with the general population. Additionally, higher levels of neuroticism, as a proposed psychological measure of stress sensitivity, increase the risk of schizophrenia. HPA axis dysregulation is one of the possible mechanisms related to the vulnerability–stress model of schizophrenia, and recent studies revealed a possible role of the functional genetic variants of FK506-binding protein 51 (FKBP5) gene which modulate activity of HPA axis. The purpose of the present study was to investigate impact of FKBP5 on schizophrenia in Serbian patients and to explore relationship between genetic variants and neuroticism by using the case–sibling–control design. In 158 subjects, we measured psychotic experiences, childhood trauma and neuroticism. Nine single-nucleotide polymorphisms (rs9295158, rs3800373, rs9740080, rs737054, rs6926133, rs9380529, rs9394314, rs2766533 and rs12200498) were genotyped. The genetic influence was modeled using logistic regression, and the relationship between genetic variants and neuroticism was assessed by linear mixed model. Our results revealed genetic main effect of FKBP5 risk alleles (A allele of rs9296158 and T allele of rs3800373) and AGTC “risk” haplotype combination (rs9296158, rs3800373, rs9470080 and rs737054, respectively) on schizophrenia, particularly when childhood trauma was set as a confounding factor. We confirmed strong relationship between neuroticism and psychotic experiences in patients and siblings and further showed relationship between higher levels of neuroticism and FKBP5 risk variants suggesting potential link between biological and psychosocial risk factors. Our data support previous findings that trauma exposure shapes FKBP5 impact on schizophrenia.

Improving Current Treatments for Schizophrenia.

Preclinical Research

Neuroticism and facial emotion recognition in healthy adults.

The aim of the present study was to examine whether healthy individuals with higher levels of neuroticism, a robust independent predictor of psychopathology, exhibit altered facial emotion recognition performance.

»Treatment Resistance« Enigma Resolved by Pharmacogenomics − A Case Study of Clozapine Therapy in Schizophrenia / Enigma »Terapo-Rezistence« Razrešena Uz Pomoć Farmakogenomike - Prikaz Slućaja Terapije Klozapinom U Shizofreniji

Summary The introduction of antipsychotic medication in the 1950s forever changed the outlook on the treatment of schizophrenia, although there is still a large proportion of patients who do not reach functional recovery. At least 30% of patients do not respond to clozapine, the tricyclic dibenzodiazepine with complex pharmacological actions, which was proven to be more effective than any other antipsychotic in the treatment of schizophrenia. According to most of the therapeutic guidelines for schizophrenia, clozapine is the third line therapy for patients who did not respond to other antipsychotics. Large inter-individual variability exists for clozapine bioavailability and plasma steadystate concentrations and clearance. Clozapine is metabolized by the cytochrome P450 oxidase enzyme family (CYP450). Cytochrome P450 1A2 (CYP1A2), which is polymorphically expressed in humans, is the main enzyme of clozapine metabolism. This case report addresses the influence of CYP1A2*1F genetic polymorphism on cloza - pine metabolism, explains the primary non-response of a young patient with schizophrenia due to increased gene expression in homozygous genotype *1F/*1F (increased metabolism of clozapine) and underlies the importance of personalizing schizophrenia treatment by means of genetic and other molecular tools, at least in the cases of »treatment resistance«. Kratak sadržaj Početak primene antipsihotika pedesetih godina prošlog veka zauvek je promenio perspektivu lečenja shizofrenije, mada značajan broj pacijenata sa ovom bolešću još uvek ne postiže funkcionalni oporavak. Klozapin, triciklični di- benzodijazepin sa kompleksnim tarmakološkim profilom, koji se u tretmanu shizofrenije pokazao efikasnijim od osta- lih antipsihotika, uglavnom je treča linija terapijskog izbora. Međutim, u najmanje 30% slučajeva obolelih od shizofreni- je stanje se ne poboljšava ni na terapiji klozapinom, pa se zbog toga pristupa polifarmaciji i odstupa od racionalne primene psihofarmaka. Postoji velika interindividualna vari- jabilnost u bioraspoloživosti klozapina, stabilnoj koncentra- ciji u plazmi i njegovom klirensu. Klozapin se metaboliše preko sistema citohrom P450 oksidaza (СYР450). Cito- hrom P450 1A2 (СYР1А2), koji se u humanoj populaciji odlikuje genskim polimorfizmima, glavni je enzim u meta- bolizmu klozapina. Aktuelni prikaz slučaja ilustruje uticaj polimorfizma CYP1A2*1F na metabolizam klozapina, gde je primarni izostanak odgovora na terapiju kod mlade paci- jentkinje sa shizofrenijom objašnjen povećanjem ekspresije gena kod homozigotnog genotipa *1F/*1F (povišen meta- bolizam klozapina), i naglašava značaj personalizacije tret- mana shizofrenije uz pomoć genetike i drugih molekularnih parametara, posebno u slučajevima »terapo-rezistence«.

O-34 - Phosphorylation of leukocyte glucocorticoid receptor as a measure of stress vulnerability in healthy women and men

Introduction The core mechanism of maladaptive stress response includs aberrant phosphorylation of the glucocorticoid receptor (GR) at serine 221 (pGR211) or GR serine 226 (pGR226), which enhance or attenuate GR transcriptional activity, respectively. In the current study we tested a hypothesis that alterations in the nuclear levels and/or ratios of GR, pGR211 and pGR226 in peripheral blood mononuclear cells (PBMC) may reflect individual level of distress and/or psychological vulnerability. Methods In 36 healthy adults we evaluated the relation of total PMBC nuclear level of GR (tGR), pGR221, pGR226 and activated JNK1 (pJNK1) and its relation to environmental stress exposure, self reported distress (Depression Anxiety Stress Scale) and personality traits (Eysenck Personality Questionnaire). Molecular and psychometric data were cross-correlated by SPSS MannWhitney statistics. Results In women, self reported distress (DASS score) correlated more with neuroticism, while in men it correlated with environmental stressor. Also, in women, PBMC nuclear tGR level correlated with pGR211, while in men it correlated with pGR226. Cross-analysis of psychometric and molecular data indicated that in women both neuroticism and self reported distress correlated positively with nuclear pGR226, while in men the only correlation was between environmental stressor and pJNK1. Conclusion This findings provide preliminary support that the nuclear level of pGR226 and pGR211/pGR226 ratio in PBMC may serve as a measure of affective vulnerability in healthy women, while nuclear level of pJNK1 may reflect environmentaly imposed distress in healthy men. The relevance of these molecular parameters as biomarkers requires broader verification in healthy and in clinical settings.

Openness to experience shortens duration of untreated psychosis in Serbian clinical population.

To determine duration of untreated psychosis (DUP) in patients with schizophrenia‐spectrum disorders from Serbia and to analyse factors that potentially contribute to the treatment delay, with focus on personality traits.

FKBP5 Epigenetic Changes In Schizophrenia: Similarity To Stress-Related Conditions

Background Hypothalamic–pituitary–adrenal (HPA) axis dysregulation is a potential neurobiological mechanism proposed by vulnerability-stress model for schizophrenia. Recent studies highlighted the role of functional genetic variants of FKBP5 gene, which affect the activity of HPA axis following stress exposure, and supported the hypothesis of increased stress sensitivity in schizophrenia. Additionally, FKBP5 demethylation in Intron 7 was observed in stress-related conditions thus the purpose of this pilot study was to investigate FKBP5 methylation levels at Intron 7 in schizophrenia. Methods Ethnically homogeneous Serbian sample of 24 schizophrenia spectrum patients and 24 controls matched by age and gender with patients’ group, were analyzed regarding DNA methylation levels at three CpG sites and average methylation level in Intron 7. Accordingly, we covered the area of GRE that has been reported to be associated with altered stress responsiveness. Epigenetic changes in FKBP5 Intron 7 were measured by Sanger sequencing and compared between the groups using t test as appropriate. Results Analyses revealed decreased FKBP5 methylation at the three targeted CpG sites (CpG1, CpG2, and CpG3) in patients compared to controls (p=0.026, p=0.017, and p=0.027, respectively). Similarly, when we averaged methylation scores of the observed region in Intron 7, significant demethylation was detected in patients (p=0.003). Discussion To the best of our knowledge this is the first study which explored FKBP5 epigenetic changes in schizophrenia. Conclusively to previous stress-related conditions, our preliminary results revealed significant decrease of FKBP5 methylation levels in Intron 7 in patients with schizophrenia. FKBP5 demethylation presents further insight into the reported FKBP5 genetic influence in psychosis and could be promising terapeutic target for the prevention of the onset and the course of schizophrenia.