[Clinical and psychopathological characteristics of patients with late-onset schizophrenia and schizophrenia-like psychoses in clusters identified by biological parameters]. / Kliniko-psikhopatologicheskie kharakteristiki bol'nykh s pozdno manifestiruyushchimi shizofreniei i shizofrenopodobnymi psikhozami v klasterakh, vydelennykh po biologicheskim parametram.

OBJECTIVE: To assess clinical and psychopathological characteristics of late-aged female patients with late-onset psychoses in clusters formed on the basis of biochemical and immunological blood parameters. MATERIAL AND METHODS: We examined 59 women with schizophrenia and schizophrenia-like psychoses with onset after 40 years (ICD-10 F20, F22.8, F25, F23, F06.2), including 34 women with late-onset (40-60 years) and 25 with very late onset psychoses (after 60 years). At the time of hospitalization, a clinical/ psychopathological study was carried out using CGI-S, PANSS, CDSS, and HAMD-17, as well as the activities of glutathione reductase (GR) and glutathione-S-transferase (GT) have been determined in erythrocyte hemolysates, and the activities of leukocyte elastase (LE) and α1-proteinase inhibitor (α1-PI) have been assessed in blood plasma. Biochemical and immunological parameters have been also determined in 34 age-matched mentally healthy women. RESULTS: Clustering by signs such as GR, GT, LE and α1-PI has yielded two clusters of objects (patients) significantly different in GT (p<0.0001), LE (p<0.0001), and α1-PI (p<0.001) activities. Relatively to the controls, in the cluster 1 patients, the activities of GST and α1-PI are increased, the activity of LE is decreased, whereas, in the cluster 2 patients, the activity of GR is decreased, and the activities of LE and α1-PI are increased. Cluster 1 patients differ from cluster 2 patients in greater severity of the condition (CGI-S, p=0.04) and higher total scores on PANSS subscales' items. Cluster 1 includes 76% of patients with very late onset. Different correlations between clinical and biological signs are found in two clusters. CONCLUSION: The identified clusters have different clinical and psychopathological characteristics. Dividing patients into subgroups according to biochemical and immunological parameters is promising for the search for differentiated therapeutic approaches.

Epidemiological Screening for the Risk of Mental, Behavioral and Developmental Disorders, Including Autism, in Early Childhood: Data for Russia 2017-2019.

We aimed to screen children aged 18-48 months in the general population of nine Russian regions for risk of mental, behavioral and developmental disorders (MBDDs) including autism spectrum disorders (ASD) using an original screening tool. The prevalence of the risk for MBDDs is 1307:10,000 (13.07%), the prevalence of clinically verified cases of MBDDs is 151:10,000 (1.51%), whereas the prevalence of ASD among them is 18:10,000 (0.18%). Basing on our results, the screening procedures are already integrated into the Russian primary care system since the end of 2019. Screening of the risk for MBDDs including ASD in Russia among children in the general pediatric population is a promising area of preventive medicine.

[Correlations between clinical features of bipolar affective disorder and serum concentrations of ANKRD12 gene product, coagulation factor XIII, and cadherin 5]. / Svyaz' kontsentratsii belka-produkta gena ANKRD12, XIII faktora koagulyatsii i kadgerina 5 v syvorotke krovi s klinicheskimi osobennostyami bipolyarnogo affektivnogo rasstroistva.

OBJECTIVE: To determine the serum concentrations of some proteins identified in the blood serum of patients with bipolar affective disorder (BD) and to evaluate the relationship of their concentrations with clinical characteristics of BD. MATERIAL AND METHODS: Protein concentrations of cadherin 5, coagulation factor XIII and ANKRD12 gene product in the blood serum of patients with BD and healthy individuals were determined using commercial ELISA kits. The severity of current depression and the main clinical manifestations of BD were assessed by SIGH-SAD and CGI upon admission to the hospital before the prescription of therapy. Data analysis was carried out using the Kruskal-Wallis and Mann-Whitney tests. RESULTS: Comparison between subgroups of BD patients, selected in accordance with the current affective episode (hypomanic, manic, depressive and mixed), and control group by the Kruskal-Wallis test has shown the significant difference in ANKRD12 protein product concentration (p=0.0448). When comparing pairwise each subgroup with the control group, the significant difference in ANKRD12 product concentration was found only in the case of a subgroup of BD patients with current depressive episode (Mann-Whitney test, p=0.006). Pairwise comparison of the ANKRD12 product concentrations in subgroups of BD patients with different disease duration (<5 years, 6-10 years, and >11 years) and in the control group by the Mann-Whitney test has revealed significant differences (p=0.04, p=0.01, p=0.02, respectively), the maximum protein concentration was found in patients with disease duration >11 years. Significant negative correlation of cadherin 5 concentration with the severity of atypical depressive symptoms assessed by SIGH-SAD (r=-0.69, p=0.038) and the decrease in cadherin 5 concentration in BD patients with greater severity of the current depressive episode were revealed. Significant negative correlation was found between factor XIII concentration and CGI scores (r=-0.62, p=0.032). CONCLUSION: The results indicate a possible link of the studied proteins with BD pathogenesis. Further study of these proteins may contribute to the development of new therapeutic and diagnostic techniques.

[Activity of enzymes of glutamate, energy and glutathione metabolism in the first juvenile depression with attenuated symptoms of schizophrenia]. / Aktivnost' fermentov glutamatnogo, energeticheskogo i glutationovogo obmena pri pervykh yunosheskikh depressiyakh s attenuirovannymi simptomami shizofrenii.

OBJECTIVE: To reveal clinical and biological correlations in patients with attenuated symptoms of schizophrenia in the first juvenile depression, namely, the correlation between SOPS and HDRS-21 scores and the levels of activities of glutathione, glutamate and energy metabolism enzymes in the blood of patients. MATERIAL AND METHODS: The study included 81 young men, aged 16-25 years, with the first depressive episode (ICD-10 items F32.1, F32.2, F32.38, F32.8), from which the groups with predominantly attenuated positive symptoms (group 1, n=36) and predominantly attenuated negative symptoms (group 2, n=24), and a group without attenuated schizophrenia symptoms (group 3, n=21) were selected. The control group consisted of 20 mentally healthy men aged 19-25 years. Psychometric methods (SOPS and HDRS-21) and psychopathological methods were used. Activities of cytochrome c oxidase (COX), glutamate dehydrogenase (GDH), glutathione reductase (GR), and glutathione S-transferase (GST) enzymes were determined spectrophotometrically in blood cells. RESULTS: When compared with the control group, activities of platelet GDH, GR, and GST (before and after treatment) were significantly reduced in groups 1, 2, 3 (Mann-Whitney U-test, p<0.0002, p<0.001 and p<0.0001, respectively); the activity of erythrocyte GST was reduced in group 1, and the activities of erythrocyte GR and GST were reduced in group 3 (p<0.05). In group 1, baseline COX (before treatment) was positively correlated with post-treatment SOPS-N scores (R=0.580, p=0.0003), while baseline erythrocyte GR was negatively correlated with post-treatment HDRS-21 scores (R=-0.591, p=0.0004). In group 2, baseline GDH levels were positively correlated with post-treatment scores on SOPS-P (R=0.425, p=0.0384), SOPS-N (R=0.500, p=0.0129), SOPS total (R=0.526, p=0.0083) and HDRS-21 (R=0.479, p=0.0180). CONCLUSION: The discovery of clinical and biological correlations in groups of patients with attenuated symptoms of schizophrenia in the structure of juvenile depression contributes to understanding the pathogenetic mechanisms of the formation of a high clinical risk of psychosis and contributes to the search for markers of the initial stages of schizophrenia.

[The link of platelet cytochrome C-oxidase activity with some clinical parameters of depression in elderly patients]. / Svyaz' aktivnosti trombotsitarnoi tsitokhrom s s-oksidazy s otdel'nymi klinicheskimi parametrami depressii u bol'nykh pozhilogo vozrasta.

OBJECTIVE: To search for correlations between platelet cytochrome c-oxidase (COX) activity and the quality of therapeutic outcomes and other clinical parameters of depression in elderly patients. MATERIAL AND METHODS: Twenty elderly women, aged 55-78 years, with depressive episodes in recurrent depressive disorder (RDD) or bipolar affective disorder (BD) were studied. COX activity and severity of depression were evaluated twice: before the beginning of antidepressant treatment and at the 28-th day of the therapy, using the Hamilton Depression Rating Scale (HAMD-17) and the Hamilton Anxiety Rating Scale (HAM-A). RESULTS: Significant correlations were revealed between platelet COX activity and some clinical parameters of the disease and the severity of depression in patients after treatment. The baseline level of the platelet COX activity was correlated with the age of disease onset (R= -0.63, p=0.003) and its duration (R=0.55, p=0.010). Significant negative correlations were also found between the baseline level of COX activity and depression severity (HAMD-17 total score) (R= -0.48, p=0.032) and the severity of anxiety (HARSHAM-A total score) (R= -0.54, p=0.010) after 28-day treatment. CONCLUSION: This pilot study has revealed a link between platelet COX activity and the severity of depression and anxiety after a 28-day antidepressant therapy. The results indicate the prospects for further study of COX as a biomarker of therapeutic outcomes in elderly patients with depression.

Decrease in 130 kDa- amyloid protein precursor protein (APP) and APP protein ratio in schizophrenia platelets.

Cognitive dysfunction is common among people with schizophrenia. The molecular substrates underlying this remain poorly understood. To address this, we analyzed changes in amyloid precursor protein (APP) in platelets of people with acute schizophrenia (n=24) and control subjects (n=20) by ECL-immunoblotting. APP bands corresponding to molecular masses of ∼130, ∼110 and ∼100 kDa, and the APP ratio (APPr: highest APP molecular mass vs lowest APP molecular mass bands) were quantified. The intensity of 130 kDa-APP and the APPr were significantly reduced in schizophrenia patients compared to control subjects. The age-associated decreases in the 130 kDa, ∼110 kDa proteins and APPr were present in patients, but not controls. Our results confirm peripheral APP metabolism is altered in people with schizophrenia. Further work is now warranted on a larger sample of diseased subjects with detailed cognitive assessment to determine the APP role in cognitive processing in schizophrenia, how it is related to severity and disease progression, as well as outcomes.

[The activity of enzymes of glutathione metabolism in blood cells of patients with a high risk of manifestation of endogenous psychoses and patients with the first psychotic episode]. / Aktivnost' fermentov glutationovogo obmena v formennykh élementakh krovi u patsientov s vysokim riskom manifestatsii éndogennykh psikhozov i bol'nykh s pervym psikhoticheskim pristupom.

AIM: To assess the activity of glutathione reductase (GR) and glutathione-S-transferase (GST) in blood cells of patients at clinical high-risk (HR) state for psychosis, in first-episode patients with schizophrenia and schizoaffective disorder (SD), and control group, and to seek correlations of these biochemical parameters with clinical assessments in patients. MATERIAL AND METHODS: The study included male patients at HR (n=21, 16-25 years old), first-episode patients with schizophrenia (F20, n=14, 18-25 years old) and SD (F25, n=20, 16-25 years old), and 12 people of the control group (19-25 years old). Psychometric scales (SOPS, HDRS, and PANSS) and psychopathological methods were employed. GR and GST enzymatic activities were determined spectrophotometrically. RESULTS: The activities of platelet GR and GST in all groups of patients both before and after treatment were lower than in controls (p<0.01). The platelet GST activity was lower in patients at HR compared to patients with schizophrenia before treatment and lower than in patients with SD after treatment (p<0.05), it was higher in patients with schizophrenia than in patients with SD before treatment (p<0.05). Erythrocyte GST activity in patients with HR was lower than in patients with SD after treatment, and in the latter it exceeded that in patients with schizophrenia and controls (p<0.05). Complex and different patterns of changes in the activities of erythrocyte and platelet GR and GST in patients with schizophrenia spectrum disorders, occurring both before the first psychotic episode in the initial stage of disease, and in the first-episode patients, were detected. CONCLUSION: The activity of glutathione-converting enzymes in endogenous psychoses of the schizophrenic spectrum, including its early stages, can be used as a biomarker for predicting the development of psychosis, the course of disease, and as criteria for evaluation of therapeutic response to antipsychotic treatment.

[Glutathione reductase and glutathione-S-transferase in blood cells in schizophrenia and schizophrenia spectrum disorders]. / Glutationreduktaza i glutation-S-transferaza v formennykh élementakh krovi pri shizofrenii i rasstroistvakh shizofrenicheskogo spektra.

AIM: To compare glutathione reductase (GR) and glutathione-S-transferase (GST) enzymatic activities in blood cells (erythrocytes and platelets) of patients with schizophrenia spectrum disorders and in the control group and to search for correlations of these biochemical parameters with clinical psychiatric assessments of the patient. MATERIAL AND METHODS: The study included patients (97 men) with schizophrenia spectrum disorder (schizophrenia and schizoaffective disorders) in an acute state of exacerbation of psychotic symptoms and 33 men without mental pathology. Symptom severity was measured with the PANSS before and after antipsychotic therapy. GR and GST activities were determined spectrophotometrically. RESULTS: There were no significant between-group differences in the activities of erythrocyte GR and GST. In platelets, the GR activity was lower in all patients' groups than in controls, whereas the GST activity in patients with schizophrenia relapses and in patients with schizoaffective disorder (SD) was lower than in controls (p<0.05) both before and after treatment. Differences between subgroups of first-episode patients (schizophrenia and SD) and patients with schizophrenia relapses were found not only in the levels of erythrocyte and platelet GR and GST activities, but also in the changes of these enzymatic activity levels under antipsychotic treatment, as well as in links binding these enzymatic activities and PANSS scores. CONCLUSION: The decreased level of GR and GST, the glutathione-dependent enzymes, contributes to the reduction of antioxidant defense in schizophrenia spectrum disorders. The correlations linking the basal levels of GR and GST activities with the results of clinical assessments after treatment allow us to consider these parameters as potential biomarkers for predicting treatment response.

Recombinant Human Erythropoietin Proteins Synthesized in Escherichia coli Cells: Effects of Additional Domains on the in vitro and in vivo Activities.

The aim of this work was to compare biological activities of three variants of bacterially expressed human recombinant erythropoietin (EPO) with additional protein domains: 6His-s-tag-EPO protein carrying the s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) at the N-terminus and HBD-EPO and EPO-HBD proteins containing heparin-binding protein domains (HBD) of the bone morphogenetic protein 2 from Danio rerio at the N- and C-termini, respectively. The commercial preparation Epostim (LLC Pharmapark, Russia) produced by synthesis in Chinese hamster ovary cells was used for comparison. The EPO variant with the C-terminal HBD domain connected by a rigid linker (EPO-HBD) possesses the best properties as compared to HBD-EPO with the reverse domain arrangement. It was ~13 times more active in vitro (i.e., promoted proliferation of human erythroleukemia TF-1 cells) and demonstrated a higher rate of association with the erythropoietin receptor. EPO-HBD also exhibited the greatest binding to the demineralized bone matrix (DBM) and more prolonged release from the DBM among the four proteins studied. Subcutaneous administration of EPO-HBD immobilized on DBM resulted in significantly more pronounced vascularization of surrounding tissues in comparison with the other proteins and DBM alone. Therefore, EPO-HBD displayed better performance with regard to all the investigated parameters than other examined EPO variants, and it seems promising to study the possibility of its medical use.

[α-Klotho protein in neurodegenerative and mental diseases]. / Belok α-kloto pri neirodegenerativnykh i psikhicheskikh zabolevaniiakh.

The review aims to attract attention of psychiatrists and neurologists to a role of α-Klotho protein in biochemical mechanisms that counteract pathogenic processes of neurodegenerative and psychiatric diseases and to possible therapeutic potential of the protein. Basing on the analysis of contemporary literature, the authors summarized the results of model experiments and a few clinical trials (in psychiatry and neurology) indicating the role of α-Klotho protein in the brain processes of neurogenesis, dendrite growth, myelination (oligodendroglia differentiation and activity), regulation of antioxidant system, and synthesis of glutamate neurotransmitter system components, regulation of the activity and synthesis of ion channel protein components and membrane transporters, synaptic plasticity. It is concluded that α-Klotho protein can be used for therapeutic purposes in diseases associated with pathological brain aging, and/or in diseases associated with insufficient synthesis of this protein.

[The activity of erythrocyte and platelet glutathione reductase and glutathione-S-transferase in paranoid schizophrenia]. / Aktivnost' éritrotsitarnykh i trombotsitarnykh glutationreduktazy i glutation-S-transferazy pri paranoidnoi shizofrenii.

AIM: A comparative evaluation of glutathione reductase (GR) and glutathione-S-transferase (GST) activities in erythrocytes and platelets of patients with schizophrenia. MATERIAL AND METHODS: Fifty patients, 47 men and 3 women, aged 25-56 years (medium 34) with acute paranoid schizophrenia (F20.0 ICD-10) with hallucinatory-paranoid or paranoid syndrome were studied. The control group consisted of 48 healthy people, 45 men and 3 women, aged 21-59 years (medium 38). GR activity was determined by the oxidation of NADP-H in the reduction reaction of oxidized glutathione. GST activity was determined by the rate of chromogenic conjugate formation between glutathione and 1-chloro-2.4-dinitrobenzene. RESULTS: No differences in the erythrocyte GR and GST activities between the control group and patients with schizophrenia were found. The platelet activity of GR and GST was significantly lower in patients compared to the control group (Mann-Whitney U test, p<0.01). Spearman rank correlation analysis showed that the erythrocyte GST activity was significantly correlated with PANSSneg scores when measured at the beginning of the study, GST was higher in those patients who had less PANSSneg scores after treatment (R=-0.41, p<0.05). The activity of platelet GST in patients with schizophrenia was correlated with the severity of positive symptoms (PANSSpos score) at the beginning of the study before taking therapy (R=-0.31, p<0.05), i.e. the more prominent psychotic symptoms were expressed in patients with lower GST activity. Upon completion of therapy, this association disappeared. CONCLUSION: The activity of glutathione-dependent enzymes in the blood cells of schizophrenic patients determined before the beginning of antipsychotic pharmacotherapy may be important for objective assessment of this metabolic system status and the degree of its impairment in patients.

Synthesis in Escherichia coli and Characterization of Human Recombinant Erythropoietin with Additional Heparin-Binding Domain.

Recombinant human erythropoietin (EPO) with additional N-terminal heparin-binding protein domain (HBD) from bone morphogenetic protein 2 was synthesized in Escherichia coli cells. A procedure for HBD-EPO purification and refolding was developed for obtaining highly-purified HBD-EPO. The structure of recombinant HBD-EPO was close to that of the native EPO protein. HBD-EPO contained two disulfide bonds, as shown by MALDI-TOF mass spectrometry. The protein demonstrated in vitro biological activity in the proliferation of human erythroleukemia TF-1 cell test and in vivo activity in animal models. HBD-EPO increased the number of reticulocytes in the blood after subcutaneous injection and displayed local angiogenic activity after subcutaneous implantation of demineralized bone matrix (DBM) discs with immobilized HBD-EPO. We developed a quantitative sandwich ELISA method for measuring HBD-EPO concentration in solution using rabbit polyclonal serum and commercial monoclonal anti-EPO antibodies. Pharmacokinetic properties of HBD-EPO were typical for bacterially produced EPO. Under physiological conditions, HBD-EPO can reversibly bind to DBM, which is often used as an osteoplastic material for treatment of bone pathologies. The data on HBD-EPO binding to DBM and local angiogenic activity of this protein give hope for successful application of HBD-EPO immobilized on DBM in experiments on bone regeneration.

[Autism spectrum disorders in children and adults: the experience of reserches from different countries]. / Rasstroistva autisticheskogo spektra u detei i vzroslykh: podkhody k probleme v raznykh stran.

Current studies on the development autism spectrum disorders (ASD) at different ages are reviewed. The review highlights the increasing interest to this problem and converging positions of researchers from different countries, encouraged by the development of international and other classifications of mental diseases, on the terminology, classification and prevalence of ASD in children. An important feature of the present stage is to draw attention to an understudied problem of ASD in adults, including elderly, and provision of medical and social care to these patients.

Recombinant Human Erythropoietin with Additional Processable Protein Domains: Purification of Protein Synthesized in Escherichia coli Heterologous Expression System.

Three variants of human recombinant erythropoietin (rhEPO) with additional N-terminal protein domains were obtained by synthesis in an Escherichia coli heterologous expression system. These domains included (i) maltose-binding protein (MBP), (ii) MBP with six histidine residues (6His) in N-terminal position, (iii) s-tag (15-a.a. oligopeptide derived from bovine pancreatic ribonuclease A) with N-terminal 6His. Both variants of the chimeric protein containing MBP domain were prone to aggregation under nondenaturing conditions, and further purification of EPO after the domain cleavage by enterokinase proved to be impossible. In the case of 6His-s-tag-EPO chimeric protein, the products obtained after cleavage with enterokinase were successfully separated by column chromatography, and rhEPO without additional domains was obtained. Results of MALDI-TOF mass spectrometry showed that after refolding 6His-s-tag-EPO formed a structure similar to that of one of native EPO with two disulfide bonds. Both 6His-s-tag-EPO and rhEPO without additional protein domains purified after proteolysis possessed the same biological activity in vitro in the cell culture.

[Comparative study of glutamate dehydrogenase in the brain of patients with schizophrenia and mentally healthy people]. / Glutamatdegidrogenaza v mozge bol'nykh shizofreniei i psikhicheski zdorovykh lits.

AIM: To compare the glutamate dehydrogenase (GDH) activity and amounts of GDHI, GDHII, and GDHIII immunoreactive forms in prefrontal, anterior and posterior cingulate cortex and cerebellar cortex of patients with schizophrenia and control subjects. MATERIAL AND METHODS: GDH enzymatic activity was measured and levels of GDH immunoreactive forms were determined in extracts of autopsied samples of prefrontal, anterior and posterior cingulate cortex (areas 10, 24, and 23 by Brodmann), and cerebellar cortex of patients with schizophrenia (n=8) and controls (n=9). RESULTS AND CONCLUSION: GDH enzymatic activity was significantly increased in the prefrontal cortex (area 10) (p<0.004), the posterior cingulate cortex (area 23) (p<0.05) and the cerebellar cortex (p<0.002) and was unchanged in the anterior cingulate cortex (area 24) in patients with schizophrenia compared to controls. The levels of immunoreactive GDH I, GDH II and GDH III were significantly higher in the prefrontal cortex of patients with schizophrenia than in controls (p<0.008, p<0.003, and p<0.0001, respectively). Levels of all three immunoreactive GDH forms were unchanged in the anterior cingulate cortex (area 24), but they were increased in the posterior cingulate cortex (area 23) (p<0.004, p<0.001 and p<0.02, respectively). The levels of immunoreactive GDH II and GDH III, but not GDH I, were significantly increased in the cerebellar cortex of patients with schizophrenia compared with the control group (p<0.02 and p<0.001, respectively). The alteration in the levels of GDH immunoreactive forms in the brain of patients with schizophrenia is one of the causes of impaired brain glutamate metabolism and an important aspect of schizophrenia pathogenesis.

Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) with Additional Protein Domain Synthesized in E. coli: In Vivo Osteoinductivity in Experimental Models on Small and Large Laboratory Animals.

Recombinant human bone morphogenetic protein-2 with an additional s-tag domain (s-tag-BMP-2) synthesized in E. coli is characterized by higher solubility and activity than the protein without additional s-tag domain, which increases the yield during purification and simplifies protein introduction into the osteoplastic materials. The high osteoinductivity of the demineralized bone matrix with s-tag-BMP-2 was shown on the model of regeneration of cranial defects of a critical size in mice and on the model of implantation of porous titanium matrix into defects of femoral and tibial bones in rabbits.

Klotho Protein: Its Role in Aging and Central Nervous System Pathology.

This review is devoted to Klotho protein and recent evidences for its functions in the brain. Information on transcriptional regulation of the klotho gene and posttranslational modifications of the protein resulting in multiple forms of Klotho is reviewed. Evidence is summarized that Klotho regulates the activity of protein factors, enzymes, and receptors, including data suggesting the importance of its glycosidase activity. Effects of Klotho on components of the glutamatergic neurotransmitter system, signal cascades involving protein kinases and protein phosphorylation, as well as oligodendrocyte differentiation and myelination are discussed. A possible contribution is proposed for Klotho levels in the development of central nervous system pathologies including mental disorders.

Two Variants of Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) with Additional Protein Domains: Synthesis in an Escherichia coli Heterologous Expression System.

Two variants of recombinant human bone morphogenetic protein-2 (rhBMP-2) with additional N-terminal protein domains were obtained by expression in E. coli. The N-terminal domains were s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) and lz (leucine zipper dimerization domain from yeast transcription factor GCN4). The s-tag-BMP-2 and lz-BMP-2 were purified by a procedure that excluded a long refolding stage. The resulting dimeric proteins displayed higher solubility compared to rhBMP-2 without additional protein domains. Biological activity of both proteins was demonstrated in vitro by induction of alkaline phosphatase in C2C12 cells, and the activity of s-tag-BMP-2 in vivo was shown in various experimental animal models.

[Creatine kinase isoform B distribution in the brain in schizophrenia]. / Raspredelenie izoformy V kreatinkinazy v mozge pri shizofrenii.

AIM: To compare patterns of brain isoform creatine phosphokinase (CPK B) distributions in post-mortem brain from patients with schizophrenia (Sch) and patients with somatic diseases (controls). MATERIAL AND METHODS: Extracts of readily soluble and membrane-associated proteins were prepared from post-mortem samples of prefrontal cortex (Brodmann area 10), anterior (area 24) and posterior (area 23) cingulate cortex, hippocampus and cerebellum cortex from patients with Sch and control group (the samples were matched by age and postmortem interval). CPK enzymatic activity was measured by determination of inorganic phosphate, amounts of immunoreative CPK В were estimated by ECL-Western blotting using monoclonal antibodies. RESULTS: A significant decrease in CPK activity and amounts of immunoreative CPK В was observed in fractions of readily soluble proteins in all studied brain structures of patients with Sch compared to controls (p<0.01). Significant differences in CPK activity were found in membrane-associated protein fractions from the hippocampus (p<0.01), but not from the cingulate cortex (areas 23 and 24), of Sch patients compared with controls, whereas no difference between groups was found in levels of immunoreactive CPK B in membrane-associated protein fractions from the cingulate cortex (areas 23 and 24) and hippocampus. The decrease in the amount of CPK B in the frontal cortex of patients with Sch was confirmed by purification of CPK B active dimer from brain samples of patients with Sch and controls. CONCLUSION: Changes in the levels of CPK brain isoform in the brain of patients with Sch (the decrease in CPK activity and amounts in various brain structures at different extents) lead to the substantial alteration of CPK distribution pattern among the brain areas studied, result in the disturbance of the brain energy metabolism and contribute to Sch pathogenesis.

Staphylococcus simulans Recombinant Lysostaphin: Production, Purification, and Determination of Antistaphylococcal Activity.

Staphylococcus simulans lysostaphin is an endopeptidase lysing staphylococcus cell walls by cleaving pentaglycine cross-bridges in their peptidoglycan. A synthetic gene encoding S. simulans lysostaphin was cloned in Escherichia coli cells, and producer strains were designed. The level of produced biologically active lysostaphin comprised 6-30% of total E. coli cell protein (depending on E. coli M15 or BL21 producer) under batch cultivation conditions. New methods were developed for purification of lysostaphin without affinity domains and for testing its enzymatic activity. As judged by PAGE, the purified recombinant lysostaphin is of >97% purity. The produced lysostaphin lysed cells of Staphylococcus aureus and Staphylococcus haemolyticus clinical isolates. In vitro activity and general biochemical properties of purified recombinant lysostaphin produced by M15 or BL21 E. coli strains were identical to those of recombinant lysostaphin supplied by Sigma-Aldrich (USA) and used as reference in other known studies. The prepared recombinant lysostaphin represents a potential product for development of enzymatic preparation for medicine and veterinary due to the simple purification scheme enabling production of the enzyme of high purity and antistaphylococcal activity.