Coping styles do not interact with the association between childhood trauma history and the immune-inflammatory phenotype of schizophrenia: Findings from a cross-sectional study.

Adverse childhood experiences (ACEs) are a well-known risk factor of schizophrenia. Moreover, individuals with schizophrenia are likely to use maladaptive stress coping strategies. Although it has been reported that a history of ACEs might be associated with a pro-inflammatory phenotype in patients with schizophrenia, the interacting effect of coping styles on this association has not been tested so far. In the present study, we aimed to investigate the levels of immune-inflammatory markers in patients with schizophrenia and healthy controls (HCs), taking into consideration a history of ACEs and coping strategies. Participants included 119 patients with schizophrenia and 120 HCs. Serum levels of 26 immune-inflammatory markers were determined. A history of any categories of ACEs was significantly more frequent in patients with schizophrenia. Moreover, patients with schizophrenia were significantly more likely to use emotion-focused coping and less likely to use active coping strategies compared to HCs. The levels of interleukin(IL)-6, RANTES, and tumor necrosis factor-α (TNF-α), appeared to be elevated in patients with schizophrenia after adjustment for potential confounding factors in all tested models. Participants reporting a history of any ACEs had significantly higher levels of TNF-α and IL-6. No significant main and interactive effects of active strategies as the predominant coping on immune-inflammatory markers with altered levels in patients with schizophrenia were found. Findings from the present study indicate that ACEs are associated with elevated TNF-α and IL-6 levels regardless of schizophrenia diagnosis and predominant coping styles.

Functional evidence for two distinct mechanisms of action of progesterone and selective progesterone receptor modulator on uterine leiomyomas.

OBJECTIVE: To study the specific mechanisms through which progesterone and selective progesterone receptor modulators impact the growth, synthesis, and accumulation of the extracellular matrix in uterine leiomyomas. DESIGN: Laboratory study. SETTING: Academic Research Institutions. PATIENTS (S): This study involved reproductive-age women diagnosed with infertility associated uterine leiomyomas who underwent myomectomy either after selective progesterone receptor modulator ulipristal acetate (UA) treatment or without any pharmacological pretreatment. Control samples included healthy myometrium tissue (n = 100). Specimens were obtained from the Department of Reproduction and Gynecological Endocrinology and Biobank, Medical University of Bialystok, Poland. INTERVENTIONS: Daily (5 mg/d) UA treated for 2 months (n = 100) and untreated (n = 150) patients with uterine leiomyomas or normal healthy myometrium (n = 100) tissue samples immediately after surgery were collected for transcriptional analysis and assessments. MAIN OUTCOME MEASURES: Progesterone-induced activation of the signaling pathways related to uterine leiomyomas extracellular matrix synthesis, deposition, and growth, as well as the expression profile of progesterone receptors in uterine leiomyomas, were assessed. RESULTS: The results indicated that progesterone activated the transforming growth factor-ß and SMAD3 signaling pathways and promoted proliferation, growth, and extracellular matrix remodeling in uterine leiomyomas by up-regulating SMAD3, transforming growth factor-ß (TGF-ß) receptor type 1 and II, Ras homolog A, vascular endothelial growth factor, or increasing the fibrosis-related gene collagen, type I, ɑ-1, and procollagen, type I, ɑ-1 production. In contrast, UA had inhibitory effects on these processes. The study also showed that both nuclear and membrane progesterone receptors play distinct roles in uterine leiomyoma pathobiology. CONCLUSIONS: We showed that both nuclear and membrane progesterone receptors were relevant in the treatment of uterine leiomyomas, especially when combined with selective progesterone receptor modulators. Novel therapeutic approaches combining selective progesterone receptor modulators with or without direct and indirect extracellular matrix targeting through selected specifically TGF-ß and SMAD3 (SMAD3, TGF-ß receptor types 1 and II, Ras homolog A, vascular endothelial growth factor, collagen, type I, ɑ-1) signaling pathways could therefore be a treatment option for uterine leiomyomas.

Associations of gut microbiota alterations with clinical, metabolic, and immune-inflammatory characteristics of chronic schizophrenia.

The present study had the following aims: 1) to compare gut microbiota composition in patients with schizophrenia and controls and 2) to investigate the association of differentially abundant bacterial taxa with markers of inflammation, intestinal permeability, lipid metabolism, and glucose homeostasis as well as clinical manifestation. A total of 115 patients with schizophrenia during remission of positive and disorganization symptoms, and 119 controls were enrolled. Altogether, 32 peripheral blood markers were assessed. A higher abundance of Eisenbergiella, Family XIII AD3011 group, Eggerthella, Hungatella, Lactobacillus, Olsenella, Coprobacillus, Methanobrevibacter, Ligilactobacillus, Eubacterium fissicatena group, and Clostridium innocuum group in patients with schizophrenia was found. The abundance of Paraprevotella and Bacteroides was decreased in patients with schizophrenia. Differentially abundant genera were associated with altered levels of immune-inflammatory markers, zonulin, lipid profile components, and insulin resistance. Moreover, several correlations of differentially abundant genera with cognitive impairment, higher severity of negative symptoms, and worse social functioning were observed. The association of Methanobrevibacter abundance with the level of negative symptoms, cognition, and social functioning appeared to be mediated by the levels of interleukin-6 and RANTES. In turn, the association of Hungatella with the performance of attention was mediated by the levels of zonulin. The findings indicate that compositional alterations of gut microbiota observed in patients with schizophrenia correspond with clinical manifestation, intestinal permeability, subclinical inflammation, lipid profile alterations, and impaired glucose homeostasis. Subclinical inflammation and impaired gut permeability might mediate the association of gut microbiota alterations with psychopathological symptoms and cognitive impairment.

Gut microbiota alterations in schizophrenia might be related to stress exposure: Findings from the machine learning analysis.

Specific mechanisms underlying gut microbiota alterations in schizophrenia remain unknown. We aimed to compare gut microbiota between patients with schizophrenia and controls, taking into consideration exposure stress across lifespan, dietary habits, metabolic parameters and clinical manifestation. A total of 142 participants, including 89 patients with schizophrenia and 52 controls, were recruited. Gut microbiota were analyzed using the 16 S rRNA sequencing. Additionally, biochemical parameters related to glucose homeostasis, lipid profile and inflammation were assessed. Increased abundance of Lactobacillus and Limosilactobacillus as well as decreased abundance of Faecalibacterium and Paraprevotella were found in patients with schizophrenia. The machine learning analysis demonstrated that between-group differences in gut microbiota were associated with psychosocial stress (a history of childhood trauma, greater cumulative exposure to stress across lifespan and higher level of perceived stress), poor nutrition (lower consumption of vegetables and fish products), lipid profile alterations (lower levels of high-density lipoproteins) and cognitive impairment (worse performance of attention). Our findings indicate that gut microbiota alterations in patients with schizophrenia, including increased abundance of lactic acid bacteria (Lactobacillus and Limosilactobacillus) and decreased abundance of bacteria producing short-chain fatty acids (Faecalibacterium and Paraprevotella) might be associated with exposure to stress, poor dietary habits, lipid profile alterations and cognitive impairment.

The deficit subtype of schizophrenia is associated with a pro-inflammatory phenotype but not with altered levels of zonulin: Findings from a case-control study.

There is evidence that subclinical inflammation and increased gut permeability might be involved in the pathophysiology of schizophrenia. Less is known about these phenomena in patients with the deficit subtype of schizophrenia (D-SCZ) characterized by primary and enduring negative symptoms. Therefore, in the present study we aimed to compare the levels of zonulin (the marker of gut permeability) and immune-inflammatory markers in patients with D-SCZ, those with non-deficit schizophrenia (ND-SCZ) and healthy controls (HCs). A total of 119 outpatients with schizophrenia and 120 HCs were enrolled. The levels of 26 immune-inflammatory markers and zonulin were determined in serum samples. The following between-group differences were significant after adjustment for multiple testing and the effects of potential confounding factors: 1) higher levels of interleukin(IL)- 1ß and C-reactive protein (CRP) in patients with D-SCZ compared to those with ND-SCZ and HCs; 2) higher levels of tumor necrosis factor-α and RANTES in both groups of patients with schizophrenia compared to HCs and 3) higher levels of IL-17 in patients with D-SCZ compared to HCs. No significant between-group differences in zonulin levels were found. Higher levels of IL-1ß and CRP were associated with worse performance of attention after adjustment for age, education and chlorpromazine equivalents. Also, higher levels of IL-1ß were correlated with greater severity of negative symptoms after adjustment for potential confounding factors. In conclusion, individuals with D-SCZ are more likely to show subclinical inflammation. However, findings from the present study do not support the hypothesis that this phenomenon is secondary to increased gut permeability.

The Self-Evaluation of Negative Symptoms in Differentiating Deficit Schizophrenia: The Comparison of Sensitivity and Specificity with Other Tools.

INTRODUCTION: Psychometric properties of the Self-evaluation of Negative Symptoms (SNS) in subjects with the deficit subtype of schizophrenia (SCZ-D) have not been investigated so far. This study had the following aims: (1) to assess psychometric properties of SNS in subjects with SCZ-D and (2) to explore the usefulness of SNS, in comparison with other clinical characteristics, in screening for SCZ-D. METHODS: Participants were 82 stable outpatients with schizophrenia, including 40 individuals with SCZ-D and 42 individuals with the non-deficit subtype (SCZ-ND). RESULTS: Internal consistency was acceptable-to-good in both groups. Factor analysis revealed two dimensions (apathy and emotional). There were significant positive correlations of the SNS total score with the subscore of negative symptoms from the Positive and Negative Syndrome Scale (PANSS) and significant negative correlations with scores of the Social and Occupational Functioning Assessment Scale (SOFAS) in both groups, indicating good convergent validity. The following measures were found to be appropriate screening tools for differentiating SCZ-D and SCZ-ND (p < 0.001): the SNS total score (area under the curve [AUC]: 0.849, cut-off ≥16, sensitivity: 80.0%, specificity: 78.6%), the PANSS subscore of negative symptoms (AUC: 0.868, cut-off ≥11, sensitivity: 90.0%, specificity: 78.6%), and the SOFAS (AUC: 0.779, cut-off ≤59, sensitivity: 69.2%, specificity: 82.5%). Also, adding the SOFAS (cut-off ≤59) to the SNS (cut-off: ≥16) further improved sensitivity and specificity (AUC: 0.898, p < 0.001, sensitivity = 87.5%, specificity = 82.2%). Cognitive performance and age of psychosis onset were not found to be suitable measures for differentiating SCZ-D and SCZ-ND. CONCLUSION: The present findings indicate that the SNS has good psychometric properties in subjects with SCZ-D and those with SCZ-ND. Moreover, the SNS, the PANSS, and the SOFAS might be used as screening tools for SCZ-D.

Functional Implications of Estrogen and Progesterone Receptors Expression in Adenomyosis, Potential Targets for Endocrinological Therapy.

Adenomyosis is a common gynaecological disease associated with the presence of endometrial lesions in the uterine myometrium. Estrogens have been proven to be the crucial hormones driving the growth of adenomyosis. Little is known about the distinct mechanisms of progesterone action in adenomyosis. Hence, in this study, we decided to characterize the expression of all nuclear and membrane estrogen and progesterone receptors. Additionally, as a functional investigation, we monitored prolactin production and cell proliferation after estradiol and progesterone treatments. We confirmed the presence of all nuclear and membrane estrogen and progesterone receptors in adenomyotic lesions at gene and protein levels. The expression of membrane progesterone receptors α and ß (mPRα, mPRß) as well as estrogen receptor ß (ERß) was upregulated in adenomyosis compared to normal myometrium. Estradiol significantly increased adenomyotic cell proliferation. Progesterone and cAMP upregulated prolactin secretion in adenomyosis in the same pattern as in the normal endometrium. In the present study, we showed the functional link between estradiol action and adenomyotic cell proliferation, as well as progesterone and prolactin production. Our findings provide novel insights into the sex steroid receptor expression pattern and potential regulated pathways in adenomyosis, suggesting that all receptors play an important role in adenomyosis pathophysiology.

Trauma Disrupts Reinforcement Learning in Rats-A Novel Animal Model of Chronic Stress Exposure.

Trauma, as well as chronic stress that characterizes a modern fast-paced lifestyle, contributes to numerous psychopathologies and psychological problems. Psychiatric patients with traumas, as well as healthy individuals who experienced traumas in the past, are often characterized by diminished cognitive abilities. In our protocol, we used an animal model to explore the influence of chronic trauma on cognitive abilities and behavior in the group of 20 rats (Rattus norvegicus). The experimental group was introduced to chronic (12 consecutive days) exposure to predator odor (bobcat urine). We measured the reinforcement learning of each individual before and after the exposition via the Probabilistic Selection Task (PST) and we used Social Interaction Test (SIT) to assess the behavioral changes of each individual before and after the trauma. In the experimental group, there was a significant decrease in reinforcement learning after exposure to a single trauma (Wilcoxon Test, p = 0.034) as well as after 11 days of chronic trauma (Wilcoxon-test, p = 0.01) in comparison to pre-trauma performance. The control group, which was not exposed to predator odor but underwent the same testing protocol, did not present significant deterioration in reinforcement learning. In cross-group comparisons, there was no difference between the experimental and control group in PST before odor protocol (U Mann-Whitney two-sided, p = 0.909). After exposure to chronic trauma, the experimental group deteriorated in PST performance compared to control (U Mann-Whitney Two-sided, p = 0.0005). In SIT, the experimental group spent less time in an Interaction Zone with an unfamiliar rat after trauma protocol (Wilcoxon two-sided test, p = 0.019). Major strengths of our models are: (1) protocol allows investigating reinforcement learning before and after exposition to chronic trauma, with the same group of rats, (2) translational scope, as the PST is displayed on touchscreen, similarly to human studies, (3) protocol delivers chronic trauma that impairs reward learning, but behaviorally does not induce full-blown anhedonia, thus rats performed voluntarily throughout all the procedures.

The Deficit Schizophrenia Subtype Is Associated with Low Adherence to the Mediterranean Diet: Findings from a Case-Control Study.

Accumulating evidence indicates that individuals with schizophrenia show poor dietary habits that might account for increased susceptibility to cardiovascular diseases in this population. However, it remains unknown whether this observation can be generalized over the whole population of individuals with schizophrenia. Therefore, in this study we aimed to investigate dietary habits, in terms of adherence to the Mediterranean diet (MD) in subjects with the deficit subtype of schizophrenia (SCZ-D), those with non-deficit subtype (SCZ-ND), and healthy controls (HCs). We recruited 45 individuals with SCZ-ND, 40 individuals with SCZ-D, and 60 HCs. Dietary habits were assessed using the Food Frequency Questionnaire-6 with a 12-month recall. Adherence to MD was decreased only in subjects with SCZ-D compared with HCs. Lower adherence to MD was associated with significantly higher levels of clinician-rated and self-reported negative symptoms (including alogia, avolition, and anhedonia). No significant correlations of adherence to MD with depressive symptoms were found. Lower adherence to MD was related to significantly higher body mass index in subjects with schizophrenia, but not in HCs. Our results indicate that poor adherence to MD is associated with a diagnosis of SCZ-D, higher severity of negative symptoms, and greater risk of developing overweight or obesity.

Confirmation Bias in the Course of Instructed Reinforcement Learning in Schizophrenia-Spectrum Disorders.

A large body of research attributes learning deficits in schizophrenia (SZ) to the systems involved in value representation (prefrontal cortex, PFC) and reinforcement learning (basal ganglia, BG) as well as to the compromised connectivity of these regions. In this study, we employed learning tasks hypothesized to probe the function and interaction of the PFC and BG in patients with SZ-spectrum disorders in comparison to healthy control (HC) subjects. In the Instructed Probabilistic Selection task (IPST), participants received false instruction about one of the stimuli used in the course of probabilistic learning which creates confirmation bias, whereby the instructed stimulus is overvalued in comparison to its real experienced value. The IPST was administered to 102 patients with SZ and 120 HC subjects. We have shown that SZ patients and HC subjects were equally influenced by false instruction in reinforcement learning (RL) probabilistic task (IPST) (p-value = 0.441); however, HC subjects had significantly higher learning rates associated with the process of overcoming cognitive bias in comparison to SZ patients (p-value = 0.018). The behavioral results of our study could be hypothesized to provide further evidence for impairments in the SZ-BG circuitry; however, this should be verified by neurofunctional imaging studies.

Endocannabinoid system in trauma and psychosis: distant guardian of mental stability.

Central endocannabinoid system (eCBS) is a neuromodulatory system that inhibits potentially harmful, excessive synaptic activation. Endocannabinoid receptors are abundant among brain structures pivotal in different mental disorders development (for example, hippocampus, amygdala, medial-prefrontal cortex, hypothalamus). Here, we review eCBS function in etiology of psychosis, emphasizing its role in dealing with environmental pressures such as traumatic life events. Moreover, we explore eCBS as a guard against hypothalamic-pituitary-adrenal axis over-activation, and discuss its possible role in etiology of different psychopathologies. Additionally, we review eCBS function in creating adaptive behavioral patterns, as we explore its involvement in the memory formation process, extinction learning and emotional response. We discuss eCBS in the context of possible biomarkers of trauma, and in preclinical psychiatric conditions, such as at-risk mental states and clinical high risk states for psychosis. Finally, we describe the role of eCBS in the cannabinoid self-medication-theory and extinction learning.

The Role of Dopaminergic Genes in Probabilistic Reinforcement Learning in Schizophrenia Spectrum Disorders.

Schizophrenia spectrum disorders (SZ) are characterized by impairments in probabilistic reinforcement learning (RL), which is associated with dopaminergic circuitry encompassing the prefrontal cortex and basal ganglia. However, there are no studies examining dopaminergic genes with respect to probabilistic RL in SZ. Thus, the aim of our study was to examine the impact of dopaminergic genes on performance assessed by the Probabilistic Selection Task (PST) in patients with SZ in comparison to healthy control (HC) subjects. In our study, we included 138 SZ patients and 188 HC participants. Genetic analysis was performed with respect to the following genetic polymorphisms: rs4680 in COMT, rs907094 in DARP-32, rs2734839, rs936461, rs1800497, and rs6277 in DRD2, rs747302 and rs1800955 in DRD4 and rs28363170 and rs2975226 in DAT1 genes. The probabilistic RL task was completed by 59 SZ patients and 95 HC subjects. SZ patients performed significantly worse in acquiring reinforcement contingencies during the task in comparison to HCs. We found no significant association between genetic polymorphisms and RL among SZ patients; however, among HC participants with respect to the DAT1 rs28363170 polymorphism, individuals with 10-allele repeat genotypes performed better in comparison to 9-allele repeat carriers. The present study indicates the relevance of the DAT1 rs28363170 polymorphism in RL in HC participants.

Epigenetic mechanisms, trauma, and psychopathology: targeting chromatin remodeling complexes.

Environmental pressure affects the genotype throughout different epigenetic processes. There is currently ample evidence on the role of epigenetics in developing various mental disorders. A burden of environmental pressure, such as psychological trauma, and its influence on genotype can lead to a variety of psychopathologies. Thus, this study focuses on the epigenetic activity of the complex protein machinery operating on chromatin - the ATP-dependent chromatin remodeling complexes. Although there are several recent studies on the molecular structure, functions, and taxonomy of ATP-dependent chromatin remodeling complexes, the focus of this paper is to highlight the importance of those 'protein machines' in developing psychiatric disorders. Data were obtained from human preclinical and clinical studies. The results of this review indicate an importance of ATP-dependent chromatin remodeling complexes in the interaction between environmental factors, including traumatic events, and genetic vulnerability to stress. Several studies indicate that ATP-dependent chromatin remodeling complexes play a crucial role in the development and consolidation of memory, in neurodevelopmental processes, and in etiology depressive-like behavior. Thus, the activity of those 'protein machines' emerges as a key factor in the pathophysiology of various psychiatric diseases. It can also be concluded that the limitations of clinical studies may be explained by inappropriate laboratory methods and research paradigms due to the delayed timeframe of biochemical responses to environmental stimuli. Future research in this field may enable a better understanding of the pathophysiology of psychiatric diseases and contribute to the development of novel molecular treatment targets.

Body symmetry and reproductive hormone levels in women.

Fluctuating asymmetry (FA), a morphological marker of developmental stability, may be related to an individual's biological condition, e.g., health or fertility. The aim of this study was to test if the level of a woman's FA was related to her fertility and reproductive potential as measured by reproductive hormone levels. Fifty-three healthy, non-pregnant, naturally cycling women (mean age = 23.42, SD = 1.85 years), participated in the study, conducted in Wroclaw (Poland) in May 2015. Early-follicular phase serum levels of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) were measured. FA was calculated based on anthropometric measures of six bilateral body traits, and the composite FA index was used in statistical analyses. No relationship was observed between FA and the levels of FSH, LH, and AMH (p > .05), controlled for potential confounders. However, the level of E2 was positively correlated with FA (p < .05). Thus, in young women, FA was not related to hormones levels related to ovarian reserve, but more symmetrical women had lower E2 levels. As FA is an index of developmental stability, environmental, and genetic stress, the results of the study confirm previous research suggesting that developmental conditions may be related to women's endogenous estrogen levels.

Toward a unified theory of childhood trauma and psychosis: A comprehensive review of epidemiological, clinical, neuropsychological and biological findings.

There is a growing body of research focused on the relationship between childhood trauma and the risk of developing psychosis. Numerous studies, including many large-scale population-based studies, controlling for possible mediating variables, provide persuasive evidence of a dose-response association and are indicative of a causal relationship. Existing evidence supports the specificity model, showing differential associations between particular adversities and clinical symptoms, with cumulative adversity causing less favorable clinical and functional outcomes in psychotic patients. To date, several psychological and biological models have been proposed to search for underlying developmental trajectories leading to the onset of psychosis, influencing psychopathological manifestation and negative functional outcomes due to a history of childhood trauma. In this article, we provide a unified review on the relationship between childhood trauma and psychosis by integrating results of epidemiological, clinical, neuropsychological and biological studies. The question whether psychosis with a positive history of childhood trauma should be considered as a new psychotic phenotype, requiring specific therapeutic interventions, warrants further investigation.

Oocyte zona pellucida and meiotic spindle birefringence as a biomarker of pregnancy rate outcome in IVF-ICSI treatment.

OBJECTIVES: IVF-ICSI procedures are accompanied by a continuous search for predictors of ART outcome. The properties of zona pellucida (ZP) have been believed to reflect the history of oocyte cytoplasmic maturation. The meiotic spindle (MS) is crucial for chromosomal alignment and proper separation of the maternal chromosomes. There is data suggesting that birefringent ZP and MS can clinically predict the oocyte quality and developmental potential of an embryo. The aim of the study was to examine the possible effect of ZP birefringent properties and MS visualization and localization as valuable predictors of IVF-ICSI effectiveness. MATERIAL AND METHODS: The prospective study was performed during a 16-month period. A total of 51 patients undergoing in vitro fertilization--embryo transfer (IVF-ET) treatment procedure with intracytoplasmic sperm injection (ICSI) were included. Controlled ovarian hyperstimulation (COH) was done using either a long n = 32 (62.75%) or an antagonist protocol n = 19. In the group of the 48 examined patients (aged 25-40), 46 ET were performed, resulting in 24 positive pregnancy tests and 19 (39.59%) clinical pregnancies. Oocytes were examined as follows: ZP birefringence autoscoring (OCTAX PolarAIDE), numeral autoscoring, thickness and clinical evaluation; MS visualization, if MS was visualized, localization of MS in relation to the polar body (PB). RESULTS: On day 3, 64.3% of the embryos were of good and 40.3% were of top quality. Visible differences, not statistically significant, were observed in the numeral score of ZP between oocytes selected and non-selected for ET. In cases when embryos were not of good or top quality, ZP score was higher (p = 0.005 p = 0.001). ZP manual evaluation indicated significantly stronger birefringence when pregnancy was not achieved (p = 0.022). The rate of MS positive oocytes was the highest in the group with pregnancy but it did not reach statistical significance (p = 0.471). The MS localization in relation to the PB was in most oocytes very close (< 45 degrees) in 70.9% and not different in the studied groups. CONCLUSIONS: Unexpected polarization microscopy imaging and rating of ZP and MS cannot be a direct predictor of the IVF outcome.

Amino and chlorambucil analogues of pentamidine--synthesis and biological examinations.

The amino analogues of pentamidine with a polymethylene (n = 3 - 6) chain and their chlorambucil derivatives were synthesized. The obtained compounds revealed cytotoxic effect on MCF-7 human breast cancer cell line (IC50 = 22 - 95 +/- 2 pM), mainly by the induction of apoptosis. The topoisomerase I/II inhibition assay and the ethidium displacement assay with the use of pBR322 plasmid DNA were used to the study of mechanism by which the obtained compounds could act. All the compounds are able to bind with DNA and interfere in vitro with the activity of topoisomerase (I and II). The determination of association constants with the use of calf thymus DNA, T4 coliphage DNA, poly(dA-dT)2 and poly(dG-dC)2 showed that the tested compounds bind within minor groove of B-DNA, but not selectively. The alkylating activity of chlorambucil derivatives determined in vitro using a Preussmann test was similar to the activity of chlorambucil. The influence of all the compounds on the amidolytic activity of plasmin and trypsin was also examined. The plasmin activity was inhibited by pentamidine, chlorambucil and aromatic bis-amines (IC50 = 0.1 - 8 mM), whereas the trypsin activity was influenced only by pentamidine.

[The pirymethamine influence on hexosoaminidase gene expression in synovial cell culture--preliminary report]. / Wplyw pirymetaminy na ekspresje genów heksozoaminidazy w synowiocytach in vitro--doniesienie wstepne.

UNLABELLED: Inflammation process is leading to increasing of synovial fluid and value of its pressure. Moreover, the impairment of vascular flow within synovial membrane and increased permeability of blood vessels were described. The activity of lysosomal enzymes, such as N-acetyl-beta-glucosaminidase (HEX), was increased in patients with rheumatoid arthritis in comparison to health synovial fluid. It is supposed, that HEX takes part in joint destruction. The using of HEX inhibitors in synovial cell culture and evaluation of HEX mRNA expression before and after the adding of inhibitor may contribute in showing the new ways of understanding pathogenetic pathways of motion organ disorders. THE AIM of the study was to evaluate the expression of HEXA and HEXB genes in the synovial cell culture derived from human synovial inflammatory fluid obtained from patients suffered from rheumatoid arthritis. MATERIAL AND METHODS: The inflamed synovial fluid was taken from patients suffered from rheumatoid arthritis. The following solutions of potential inhibitor--pyrimethamine were used: 20 microg/ml, 10 microg/ml, 3 microg/ml and 1.5 microg/ml. Two separate control groups were established: control group 1 where only 0.6% of ethanol was added to the synovial cell culture; control group 2 where only 0.5% DMSO was added to the synovial cell culture. The relative quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) was carried out. RESULTS. The difference in HEXA and HEXB expression was observed in synoviocytes obtained in synovial cell culture. Five time higher relative HEXA expression was determined after applying 3 microg/ml of pirymethamine compared with the control 1. The highest concentration of pirymethamine (10 and 20 microg/ml) caused the least elevation of HEXA expression. The slight decreased of HEXB expression was observed under the concentration of pirymethamine: 1.3 and 3 microg/ml. CONCLUSIONS. Pyrimethamine contributes to regulating the HEX gene expression from synovial cells. The change in gene expression level is dependent on the concentration of the pirymethamine. Our preliminary data don't let us establish the concentration of pyrimethamine that may significantly inhibit HEXA and HEXB expression. Further study may be conducted to put new insight into the pathogenesis of joint destruction in the course of rheumatoid arthritis.

Analogues of distamycin--synthesis and biological evaluation of new aromatic oligopeptides, potential anticancer agents.

Six new aromatic oligopeptides were synthesized and evaluated for their activity in the standard cell line of the mammalian tumor MCF-7 as well as in a cell-free system employing the topoisomerase I/II inhibition assay.