Clinical and pharmacological factors influencing serum clozapine and norclozapine levels.

Background: Clozapine (CLO) is a very effective antipsychotic, whose use is associated with dose-dependent risk of complications. Due to high interindividual variability in CLO metabolism, there is a need to identify factors affecting the blood concentrations of CLO and its active metabolite, norclozapine (NCLO). Methods: A total of 446 blood samples (collected from 233 women and 213 men, aged from 18 to 77 years) were included in this study and analyzed for CLO and NCLO concentrations. The patients were treated at a psychiatric hospital in Warsaw in the years 2016-2021. Serum CLO and NCLO concentrations were determined with high-performance liquid chromatography coupled to UV. Results: The following factors were shown to increase serum CLO and NCLO levels: higher CLO dose (p < 0.001), female sex (p < 0.001), nonsmoker status (p < 0.001), the use of more than two additional psychotropic drugs (only in the case of CLO; p = 0.046), concomitant use of beta-blockers (for CLO p = 0.049; for NCLO p < 0.001), and older age (for CLO p < 0.001; for NCLO p = 0.011). Despite the use of CLO at daily doses within the recommended range (200-450 mg), the evaluated serum CLO and NCLO levels were within the therapeutic ranges in only 37% and 75% of cases, respectively, with 5.6% of cases exceeding the CLO toxicity threshold. Discussion: The use of CLO at recommended doses does not guarantee achieving therapeutic concentrations of CLO or NCLO. Women and nonsmokers were at the highest risk of having toxic CLO levels.

Depression, anxiety, insomnia and interleukins in the early postpartum period.

Background: Some new mothers have been shown to suffer from anxiety and depression associated with insomnia during the postpartum period. Our study assessed the impact of demographic, psychopathological, and biochemical factors on the incidence of depression in women during the early postpartum period. Methods: A total of 119 women were evaluated at 24-48 h postpartum with the following psychometric scales: Hamilton Depression Rating Scale (HDRS), Edinburgh Postnatal Depression Scale (EPDS), Hamilton Anxiety Rating Scale (HARS) and Athens Insomnia Scale (AIS). In addition, blood was drawn to assay interleukin 6 (IL-6) and interleukin 10 (IL-10). Results: The factors that had the greatest impact on the risk of postpartum depression detected with the HDRS were high HARS scores and evidence of insomnia in the AIS. There were no significant differences in IL-6 or IL-10 levels in women with and without depression (based on either HDRS or EPDS scores) and insomnia (based on AIS) after childbirth. Considering demographic factors, divorced and single women were shown to be at higher risk of postpartum depression (based on EPDS scores). Limitations: Small sample size and short observation span. Conclusion: This study highlights the relationship between postpartum depression and both anxiety and insomnia and emphasises the importance to assess symptoms of anxiety and sleep quality as part of screening in women at risk of postpartum depression.

Factors Contributing to Risk of Persistence of Positive and Negative Symptoms in Schizophrenia during Hospitalization.

The aim of the study was to evaluate factors that may contribute to the persistence of positive, negative and other psychopathological symptoms of schizophrenia. All patients were treated in general psychiatric wards between January 2006 and December 2017. The initial study sample comprised of the medical reports of 600 patients. The main, specified inclusion criterion for the study was schizophrenia as a discharge diagnosis. Medical reports of 262 patients were excluded from the study due to no neuroimaging scans being available. The symptoms were categorised into three groups: positive, negative, and other psychopathological symptoms. The statistical analysis comprised modalities such as demographic data, clinical symptoms, as well as neuroimaging scans linking them to a potential impact of sustaining the mentioned groups of symptoms during the period of hospitalization. The analysis revealed that statistically significant risk factors of persistence of the three groups of symptoms are the elderly age, the increasing toll of hospitalizations, suicidal attempts in medical history, a family history of alcohol abuse, the presence of positive, negative and other psychopathological symptoms on admission to the hospital, as well as the absence of cavum septi pellucidi (CSP). The study showed that addiction to psychotropic drugs and a family history of schizophrenia were more frequent in patients with persistent CSP.

The cancer-risk variant frequency among Polish population reported by the first national whole-genome sequencing study.

Introduction: Population-based cancer screening has raised many controversies in recent years, not only regarding the costs but also regarding the ethical nature and issues related to variant interpretation. Nowadays, genetic cancer screening standards are different in every country and usually encompass only individuals with a personal or family history of relevant cancer. Methods: Here we performed a broad genetic screening for cancer-related rare germline variants on population data from the Thousand Polish Genomes database based on 1076 Polish unrelated individuals that underwent whole genome sequencing (WGS). Results: We identified 19 551 rare variants in 806 genes related to oncological diseases, among them 89% have been located in non-coding regions. The combined BRCA1/BRCA2 pathogenic/likely pathogenic according to ClinVar allele frequency in the unselected population of 1076 Poles was 0.42%, corresponding to nine carriers. Discussion: Altogether, on the population level, we found especially problematic the assessment of the pathogenicity of variants and the relation of ACMG guidelines to the population frequency. Some of the variants may be overinterpreted as disease-causing due to their rarity or lack of annotation in the databases. On the other hand, some relevant variants may have been overseen given that there is little pooled population whole genome data on oncology. Before population WGS screening will become a standard, further studies are needed to assess the frequency of the variants suspected to be pathogenic on the population level and with reporting of likely benign variants.

Genetics, Genomics and Emerging Molecular Therapies of Pancreatic Cancer.

The number of cases of pancreatic cancers in 2019 in Poland was 3852 (approx. 2% of all cancers). The course of the disease is very fast, and the average survival time from the diagnosis is 6 months. Only <2% of patients live for 5 years from the diagnosis, 8% live for 2 years, and almost half live for only about 3 months. A family predisposition to pancreatic cancer occurs in about 10% of cases. Several oncogenes in which somatic changes lead to the development of tumours, including genes BRCA1/2 and PALB2, TP53, CDKN2A, SMAD4, MLL3, TGFBR2, ARID1A and SF3B1, are involved in pancreatic cancer. Between 4% and 10% of individuals with pancreatic cancer will have a mutation in one of these genes. Six percent of patients with pancreatic cancer have NTRK pathogenic fusion. The pathogenesis of pancreatic cancer can in many cases be characterised by homologous recombination deficiency (HRD)-cell inability to effectively repair DNA. It is estimated that from 24% to as many as 44% of pancreatic cancers show HRD. The most common cause of HRD are inactivating mutations in the genes regulating this DNA repair system, mainly BRCA1 and BRCA2, but also PALB2, RAD51C and several dozen others.

Pharmaco-Electroencephalography-Based Assessment of Antidepressant Drug Efficacy-The Use of Magnesium Ions in the Treatment of Depression.

Pharmaco-electroencephalography (pharmaco-EEG) is a technique used to assess the effects of psychotropic medications on the bioelectrical activity of the brain. The purpose of this study was to assess the treatment response with the use of the Hamilton Depression Rating Scale (HDRS) and via EEG. Over an 8-week period, we analyzed electroencephalographic tracings of 91 patients hospitalized for major depression at the Medical University of Warsaw. Thirty-nine of those patients received tricyclic antidepressants (TCAs), 35 received fluoxetine, and 17 received fluoxetine augmented with magnesium (Mg) ions. All patients had their serum drug levels monitored. The highest proportion of patients (88.2%) who showed adequate responses to treatment was observed in the fluoxetine+Mg group, whereas the lowest rates of treatment response were observed in the TCA group (58.3%). This difference was statistically significant (p = 0.029, Phi = 0.30). Our study demonstrated a relationship between achieving remission (HDRS ≤ 6 at week 8 of treatment) and obtaining a positive pharmaco-EEG profile 6 h after administration of the first dose in the group receiving fluoxetine augmented with Mg ions (p = 0.035, Phi = 0.63).

Substance Use Disorder Status Moderates the Association between Personality Traits and Problematic Mobile Phone/Internet Use.

BACKGROUND: Associations between personality traits and problematic smartphone use (PSU) among individuals with substance use disorder (SUD) have not been widely investigated. The current study aims to assess whether SUD status moderates the association between personality traits and PSU. METHODS: The study group included 151 individuals with SUD and a normative sample (NS) comprised of 554 non-SUD students. The following self-report questionnaires were used: the Mobile Phone Problem Use Scale (MPPUS-10) to assess problematic smartphone use (PSU), the Internet Addiction Test (IAT) to assess intensity of internet use, and the NEO Five-Factor Inventory (NEO-FFI) to assess Personality traits. RESULTS: SUD status moderated the association between neuroticism and openness to new experiences on PSU. That is, greater neuroticism and openness were significantly associated with more excessive PSU among the NS. In the SUD group, greater openness was a significant protective factor against PSU. Moderation results were similar when using the IAT (which was significantly correlated with MPPUS) as an outcome. CONCLUSIONS: The presence of SUD may influence how personality traits are associated with problematic mobile phone/internet use. Given that this is among one of the first studies examining this topic, findings should be replicated with additional studies.

Psychometric properties of the polish version of the Alcohol Use Disorders Identification Test (AUDIT).

BACKGROUND: Alcohol consumption is one of the risk factors associated with over 200 types of diseases, being strongly correlated with unemployment, low socioeconomic status, traffic accidents, and violence. Alcohol is also one of the main causes of premature deaths, especially among males. Early diagnosis based on screening may help to prevent the evolution from alcohol misuse to dependence by counseling or treatment. METHODS: 1024 participants took part in the study: 300 of alcohol dependents and 724 of controls (405 students and 319 patients from General Practice) (M ± SD age = 33.7 ± 15.4 years). Finally, group of 877 participants were included to the statistical analysis due to the missing data - 453 were male and 424 were female. The median AUDIT score was 17 (IQR = 11). RESULTS: Correlation analysis showed that AUDIT scores were strongly correlated with MAST (rho = 0.764, p < 0.001) and CAGE scores (rho = 759, p < 0.001). The ROC analysis resulted in an area under the curve (AUC) of 0.95 (p < 0.001; 95 % C.I.: 0.936-0.963). The cutoff point of 22 for alcohol dependence corresponded to a sensitivity of 0.893 and a specificity of 0.863. CONCLUSION: According to our results, the AUDIT scale proved to be a useful tool with high sensitivity and specificity and therefore can be used as a valid screening measure in Poland.

Adaptation and Validation of the Polish Version of the 10-Item Mobile Phone Problematic Use Scale.

Pathological use of smartphones may be the biggest non-drug addiction of the 21st century. Therefore, rapid screening tools designed for easy identification of people with problematic mobile phone use are needed. The main aim of the present study was to validate a short version of the Mobile Phone Problematic Use Scale (MPPUS-10) in the Polish population. The study comprised 640 university students aged 18-38 years. We used a self-report questionnaire that included questions regarding socio-demographic variables and Polish versions of the Mobile Phone Problem Use Scale (MPPUS-10), Mobile Phone Addiction Assessment Questionnaire (MPAAQ in Polish KBUTK), and Internet Addiction Test (IAT) by Kimberly Young. The analysis showed high reliability for the final Polish version of MPPUS-10 (Cronbach's α = 0.78) and confirmed a significant correlation between the MPPUS-10 and the MPAAQ, which was previously used in Poland (rho = 0.56; p < 0.001). Due to the poor correlation of item number 10 with other items, we suggest dropping this item and using the nine-item Polish version. Our result also confirmed significant correlation between the MPPUS-10 and the IAT (rho = 0.54; p < 0.001). The contribution of the study is the Polish validation and adaptation of the MPPUS-10 scale with confirmed psychometric values. It provides a quick and convenient screening tool to assess problematic mobile phone use. Our results also indicate the need for a revision of available diagnostic tools in Poland.

Psychometric Properties of the Polish version of the Drug-Use Disorders Identification Test.

BACKGROUND: Substance use is a growing problem worldwide, and there is a great need to develop national policies addressing prevention and treatment of substance-use disorders (SUD). However, the lack of a commonly used, valuable diagnostic tool to assess the symptoms of SUDs precludes comparison of the prevalence of drug-use problems as well as the efficacy of policy strategies applied in different countries. This study was conducted to validate one of the commonly used scales the Drug Use Disorders Identification Test (the DUDIT) for use in Polish condition. OBJECTIVES: The aims of this study were to test the reliability of the DUDIT in the Polish population, to confirm the comparative stability of the factor structure of the instrument, and to verify previously proposed DUDIT cutoff scoring. METHODS: The group of drug users comprised 127 patients aged 19-40 years (mean age 30.37, SD 6.36) with a diagnosis of SUD, while the control group consisted of 533 students aged 19-25 years (mean age 20.72, SD 1.88). All participants completed the Polish version of the DUDIT. Internal consistency of the Polish version of the DUDIT was determined, and subsequently external validation was performed. RESULTS: Analysis showed that the Polish version of the DUDIT was characterized by a good reliability based on Cronbach's α, with a value of 0.92. The between-group comparison revealed a significant difference between the control group and substance-dependent patients corresponding to a large effect size (Cohen's d = 3.27). The receiver-operating characteristic analysis, comparing the DUDIT score to the ICD-10 diagnosis of SUD, showed an optimal cutoff value of 7 points, with a sensitivity of 0.929 and a specificity of 0.974. CONCLUSION: These results constitute preliminary evidence that the Polish version of the DUDIT may be a valid and reliable screening tool for drug-use disorders in the Polish population.

Effects of Magnesium Supplementation on Unipolar Depression: A Placebo-Controlled Study and Review of the Importance of Dosing and Magnesium Status in the Therapeutic Response.

Animal studies using tests and models have demonstrated that magnesium exerts an antidepressant effect. The literature contains few studies in humans involving attempts to augment antidepressant therapy with magnesium ions. The purpose of our study was to assess the efficacy and safety of antidepressant treatment, in combination with magnesium ions. A total of 37 participants with recurrent depressive disorder who developed a depressive episode were included in this study. As part of this double-blind study, treatment with the antidepressant fluoxetine was accompanied with either magnesium ions (120 mg/day as magnesium aspartate) or placebo. During an 8-week treatment period, each patient was monitored for any clinical abnormalities. Moreover, serum fluoxetine and magnesium levels were measured, and pharmaco-electroencephalography was performed. The fluoxetine + magnesium and fluoxetine + placebo groups showed no significant differences in either Hamilton Depression Rating Scale (HDRS) scores or serum magnesium levels at any stage of treatment. Multivariate statistical analysis of the whole investigated group showed that the following parameters increased the odds of effective treatment: lower baseline HDRS scores, female gender, smoking, and treatment augmentation with magnesium. The parameters that increased the odds of remission were lower baseline HDRS scores, shorter history of disease, the presence of antidepressant-induced changes in the pharmaco-EEG profile at 6 h after treatment, and the fact of receiving treatment augmented with magnesium ions. The limitation of this study is a small sample size.

Insomnia, postpartum depression and estradiol in women after delivery.

After childbirth, women may develop symptoms of depression with the associated sleep disturbances. This study assessed the relationship between insomnia and both depression symptoms and blood estradiol levels in women during the early postpartum period. 84 patients were assessed 24-48 h after labor. The main assessment methods were the following psychometric scales: Beck Depression Inventory (BDI), Edinburgh Postnatal Depression Scale (EPDS) and Athens Insomnia Scale (AIS). Serum estradiol levels were measured using ELISA assay. Women who developed postpartum insomnia significantly more often reported insomnia during pregnancy (P = 0.001), were more likely to have suffered from depression in the past (P = 0.007) and had significantly higher BDI (P = 0.002) and EPDS (P = 0.048) scores. Our study demonstrated no significant association between Restless Legs Syndrome (RLS) during pregnancy and postpartum insomnia. The groups of women with and without postpartum RLS showed no significant differences in the incidence of postpartum insomnia. No significant differences in estradiol levels were observed in women with and without postpartum insomnia. The study showed the following factors to play a major role in development of postpartum insomnia: an increase in Beck Depression Inventory score, a history of depression and a history of insomnia during pregnancy.

An association between genetic variation in the glutamatergic system and suicide attempts in alcohol-dependent individuals.

BACKGROUND AND OBJECTIVES: Pathological alterations of glutamatergic systems were observed in neurodegenerative and psychiatric disorders. There is some evidence that this system may be involved in the genetic vulnerability to suicide. The aim of the present study was to analyze possible relationship between the GRIN2B polymorphism and suicidal behavior. We hypothesized that this genetic factor may be associated with suicide attempts in alcohol-dependent patients and with death by suicide. METHODS: To analyze the relationship between GRIN2B and suicide attempts, the selected rs2268115 polymorphism was genotyped in a sample of 345 alcohol-dependent individuals stratified by the history of suicide attempts. The second part of the study concerning suicide was based on a sample of 510 suicide victims and 450 controls. RESULTS: The frequency of rs2268115 G allele among alcohol-dependent patients with the history of suicide attempts was significantly higher than among non-suicidal alcohol-dependent individuals (OR = 1.45, p = .033). This association was more significant when analyzing alcohol-dependent patients only without co-occurring drug dependence (OR = 1.62, p = .021). The analyzed GRIN2B polymorphism was associated with a twofold increase in odds of a suicide attempt (OR = 2.01, p = .004). No relationships between rs2268115 and death by suicide were identified. DISCUSSION AND CONCLUSIONS: Our results suggest that glutamatergic system influence susceptibility to suicide attempts in alcohol-dependent individuals. Suicidal behavior and alcohol dependence may share a common etiology related to the glutamatergic system. SCIENTIFIC SIGNIFICANCE: The major contribution of the present study is a novel finding of the possible association between GRIN2B rs2268115 polymorphism and suicide attempts in alcohol-dependent individuals. (Am J Addict 2017;26:595-601).

SIMULTANEOUS ASSAY OF ARIPIPRAZOLE AND ITS ACTIVE METABOLITE IN SERUM BY HPLC.

A rapid, convenient, precise HPLC method was developed for the simultaneous determination of aripiprazole and its active metabolite dehydroaripiprazole in blood serum. The separation was carried out by RP HPLC on Symmetry C18 column (150 x 4.6 mm; 5 µm). The mobile phase was composed of acetonitrile: water (30 : 70, v/v), pH 3.0 adjusted with o-phosphoric acid. The detection was monitored at 220 nm.

COMT and BDNF Gene Variants Help to Predict Alcohol Consumption in Alcohol-dependent Patients.

BACKGROUND: The neurobiology of alcohol dependence (AD) involves alterations in neurotransmitters and the stress response. We hypothesized that an interaction between functional variants of dopaminergic and neurotrophic genes may influence drinking in AD. METHODS: The relationship between alcohol consumption and single-nucleotide polymorphisms, Val66Met in the brain-derived neurotrophic factor (BDNF), and Val158Met in the catechol-O-methyltransferase (COMT), was analyzed among 281 alcohol-dependent individuals. RESULTS: Individuals carrying both the COMT Met158Met genotype and the BDNF Val66Val genotype drank more than those with other variants of these genes (P = 0.039). Those who had a family history of AD also drank more than those without a family history (P = 0.048). Patients with both Met/Met genotype in the COMT Val158Met polymorphism and Val/Val genotype in the BDNF Val66Met polymorphism suffered from more health problems than those carrying other variants (P = 0.030) and had lower motivation to change drinking patterns (P = 0.031). CONCLUSIONS: Patients carrying both the BDNF Val66Val and COMT Met158Met variants had higher alcohol consumption. These effects may be influenced by the effects of BDNF and COMT on dopamine responses to alcohol. Motivation-enhancing strategies might benefit the group of patients identified by genotyping in this study, and also treatment aimed at reducing alcohol consumption.

Label-free quantitative mass spectrometry for analysis of protein antigens in a meningococcal group B outer membrane vesicle vaccine.

The development of a multivalent outer membrane vesicle (OMV) vaccine where each strain contributes multiple key protein antigens presents numerous analytical challenges. One major difficulty is the ability to accurately and specifically quantitate each antigen, especially during early development and process optimization when immunoreagents are limited or unavailable. To overcome this problem, quantitative mass spectrometry methods can be used. In place of traditional mass assays such as enzyme-linked immunosorbent assays (ELISAs), quantitative LC-MS/MS using multiple reaction monitoring (MRM) can be used during early-phase process development to measure key protein components in complex vaccines in the absence of specific immunoreagents. Multiplexed, label-free quantitative mass spectrometry methods using protein extraction by either detergent or 2-phase solvent were developed to quantitate levels of several meningococcal serogroup B protein antigens in an OMV vaccine candidate. Precision was demonstrated to be less than 15% RSD for the 2-phase extraction and less than 10% RSD for the detergent extraction method. Accuracy was 70 to 130% for the method using a 2-phase extraction and 90-110% for detergent extraction. The viability of MS-based protein quantification as a vaccine characterization method was demonstrated and advantages over traditional quantitative methods were evaluated. Implementation of these MS-based quantification methods can help to decrease the development time for complex vaccines and can provide orthogonal confirmation of results from existing antigen quantification techniques.

Qualitative and quantitative evaluation of Simon™, a new CE-based automated Western blot system as applied to vaccine development.

Many CE-based technologies such as imaged capillary IEF, CE-SDS, CZE, and MEKC are well established for analyzing proteins, viruses, or other biomolecules such as polysaccharides. For example, imaged capillary isoelectric focusing (charge-based protein separation) and CE-SDS (size-based protein separation) are standard replacement methods in biopharmaceutical industries for tedious and labor intensive IEF and SDS-PAGE methods, respectively. Another important analytical tool for protein characterization is a Western blot, where after size-based separation in SDS-PAGE the proteins are transferred to a membrane and blotted with specific monoclonal or polyclonal antibodies. Western blotting analysis is applied in many areas such as biomarker research, therapeutic target identification, and vaccine development. Currently, the procedure is very manual, laborious, and time consuming. Here, we evaluate a new technology called Simple Western™ (or Simon™) for performing automated Western analysis. This new technology is based on CE-SDS where the separated proteins are attached to the wall of capillary by a proprietary photo activated chemical crosslink. Subsequent blotting is done automatically by incubating and washing the capillary with primary and secondary antibodies conjugated with horseradish peroxidase and detected with chemiluminescence. Typically, Western blots are not quantitative, hence we also evaluated the quantitative aspect of this new technology. We demonstrate that Simon™ can quantitate specific components in one of our vaccine candidates and it provides good reproducibility and intermediate precision with CV <10%.

Association of IL1A, IL1B, ILRN, IL6, IL10 and TNF-α polymorphisms with risk and clinical course of multiple sclerosis in a Polish population.

Single nucleotide polymorphisms in human pro- and anti-inflammatory genes, including IL1RN VNTR (rs315952), IL1A 4845G>T (rs17561), L1B-511C>T (rs16944), IL6-174G>C (rs1800795), IL10-1082 A>G (rs 1800896) and TNFα-308G>A (rs1800629) and their impact on multiple sclerosis risk and disease progression in a Polish population were investigated. Increased risk of MS was found for IL6-174 CC homozygotes (OR, 2.88; p<0.00001). In turn, IL1A 4845 TT genotype determined earlier appearance of MS onset whereas IL1B-511 TT genotype was associated with later occurrence of MS but faster disability progression.

Isolation and characterization of IgG1 with asymmetrical Fc glycosylation.

N-glycosylation of immunoglobulin G (IgG) at asparigine residue 297 plays a critical role in antibody stability and immune cell-mediated Fc effector function. Current understanding pertaining to Fc glycosylation is based on studies with IgGs that are either fully glycosylated [both heavy chain (HC) glycosylated] or aglycosylated (neither HC glycosylated). No study has been reported on the properties of hemi-glycosylated IgGs, antibodies with asymmetrical glycosylation in the Fc region such that one HC is glycosylated and the other is aglycosylated. We report here for the first time a detailed study of how hemi-glycosylation affects the stability and functional activities of an IgG1 antibody, mAb-X, in comparison to its fully glycosylated counterpart. Our results show that hemi-glycosylation does not impact Fab-mediated antigen binding, nor does it impact neonatal Fc receptor binding. Hemi-glycosylated mAb-X has slightly decreased thermal stability in the CH2 domain and a moderate decrease (∼20%) in C1q binding. More importantly, the hemi-glycosylated form shows significantly decreased binding affinities toward all Fc gamma receptors (FcγRs) including the high-affinity FcγRI, and the low-affinity FcγRIIA, FcγRIIB, FcγRIIIA and FcγRIIIB. The decreased binding affinities to FcγRs result in a 3.5-fold decrease in antibody-dependent cell cytotoxicity (ADCC). As ADCC often plays an important role in therapeutic antibody efficacy, glycosylation status will not only affect the antibody quality but also may impact the biological function of the product.

BDNF A196G and C270T gene polymorphisms and susceptibility to multiple sclerosis in the Polish population. Gender differences.

Two single nucleotide polymorphisms of the human BDNF gene, G196A and C270T, and their impact on the susceptibility to MS and disease progression in the Polish population were investigated. Increased risk of the disease was found for 196G/G carriers (OR -- 1.63, p=0.01) (only females) and 270C/T carriers (OR -- 7.76, p<0.001) (both males and females). The first signs of the disease appeared earlier in 196G/G than 196G/A patients (p=0.01), but it was limited to males. Our results show that C270T and G196A BDNF polymorphisms may affect susceptibility to and onset of MS, but further verification is needed, with special attention to gender differences.