[The cellular factors of innate immunity in nonpsychotic patients at high risk for schizophrenia]. / Kletochnye faktory estestvennogo immuniteta u bol'nykh s nepsikhoticheskimi psikhicheskimi rasstroistvami, otrazhayushchimi vysokii risk razvitiya shizofrenii.

AIM: Changes in the parameters of innate immunity in patients with schizophrenia are observed already in the first episode. The study was performed to find out whether these changes take place prior to disease manifestation, and what role do they play in the pathogenesis of schizophrenia. MATERIAL AND METHODS: Thirty-five male nonpsychotic patients at high risk of psychosis, aged between 17 to 23 years, were examined. Phagocyte activity (PA) of neutrophils in the blood serum was evaluated by the number of active neutrophils, i.e. phagocytic index (PhI), and phagocytic number (PhN), which was determined by counting latex particles absorbed with a single phagocytic cell. Cytotoxic activity of natural killer lymphocytes (NK CA) was evaluated by the number of cell targets K-562, which remained non-degraded after the contact with natural killer cells. The influence of monocytes on NKCA was determined as well. RESULTS AND CONCLUSION: Compared to controls, patients had the lower PhI level (p<0.001) which was compensated by the increase in PhN levels, and the lower NK CA level which was increased due to the influence of monocytes. Negative correlations between PhI and PhN (r= -0.83, p<0.01) and between the level of NKCA and PhI (r= -0.83, p<0.05) as well as the positive correlation between PhN and SOPS scores (r=0.69, p<0.01) were found. After treatment, there was the decreasein the severity of mental disorders (p<0.001). The level of PhAN was normalized in 61.9% of patients compared to 36.7% before treatment. After treatment, the proportion of patients with normal levels of NK CA was the same as before treatment (40 and 35%, respectively). The immune disturbances revealed in the study may play a role in the pathogenesis of the disease and have predictive value for schizophrenia.

[The activity of complement system in children with autistic spectrum disorders]. / Aktivnost' sistemy komplementa u detei s rasstroistvami autisticheskogo spektra.

AIM: To evaluate the state of complement system (CS) activity in children with autistic spectrum disorders (ASD) and children with schizophrenia on the basis of development and implementation of a new method of CS determination. MATERIAL AND METHODS: A study included 249 patients, aged from 3 to 14 years. The control group consisted of 279 age-matched children. The authors developed a method for integral evaluation of CS activity based on the changes in the death of free swimming ciliata Tetrahymena pyriformis measured with the apparatus BioLat (Moscow, Russia). The integral CS activity (T50) was estimated as the time of death of 50% of ciliata in the blood serum (serum concentration was 5%). RESULTS AND CONCLUSION: A comparative analysis of CS activity showed statistical differences in median T50 values between patients and controls (p<0.005). Based on CS activity levels, three groups of patients were determined: 1) with the levels lower than the lowest value of the control group (n=112 (39%)); 2) higher than the highest level of the control group (n=103 (36%)); intermittent between low and high values of the control group (n=72 (25%)). Significant differences in T50 between the psychotic autism group and children schizophrenia group were identified (p<0.005). The CS activity was lower in patients with ASD compared to children with schizophrenia.

[Interaction effect of serotonin transporter gene and brain-derived neurotrophic factor on the platelet serotonin content in stroke patients].

Platelet serotonin content in patients in the acute period of stroke is an important index of clinical changes during the post stroke period as well as a predictor of development of mental disorders. We studied the association between two polymorphisms (5-HTTLPR and Val66Met BDNF) and the platelet serotonin content in 47 patients with stroke. We also investigated the moderating effect of genetic variants on the association between platelet serotonin content and development of affective and anxiety disorders in stroke patients in the acute period of stroke. The interaction effect of two polymorphisms on levels of platelet serotonin was found. The lowest level was observed in patients with the diplotype LL*ValVal, the highest level--in the group of patients with the LL genotype and genotypes containing at least one copy of a Met allele. No moderating effect of genetic variants on the relationship between serotonin content and affective or anxiety disorder was found.

Analysis of serotonin transporter in human platelets by immunoblotting using site-specific antibodies.

We have produced a panel of site-specific antibodies recognizing different regions of the human serotonin transporter (SERT). This panel included: 1) monoclonal antibodies 23C5 (mAbs 23C5) to the C-terminal region (amino acid residues 597-630); 2) polyclonal antibodies (pAbs) to the N-terminal region (amino acid residues 69-83); 3) pAbs to the region (amino acid residues 86-100) in the beginning of the first transmembrane domain (TMD). The antibodies were produced using recombinant proteins and synthetic peptides (containing certain sequences of SERT) as antigens. These antibodies were purified by affinity chromatography, conjugated to horseradish peroxidase (HRP), and used for immunoblotting analysis of SERT in extracts of human platelets. Sodium dodecyl sulfate extracts were prepared under conditions preventing non-specific proteolytic degradation of the proteins. In platelet extracts, all antibodies were able to detect the 67 kD protein, apparently corresponding to full-length SERT molecule (its theoretical mass is about 70 kD). These antibodies also detected several polypeptides of smaller size (56, 37, 35, 32, 22, and 14 kD), apparently corresponding to N-terminal, C-terminal, and non-terminal SERT fragments. Specificity of immunostaining was confirmed by preincubation of HRP-labeled anti-SERT antibodies with excess of corresponding antigen, which resulted in disappearance of protein band staining. It is suggested that SERT undergoes a programmed proteolytic cleavage (processing) resulting in formation of several SERT-derived polypeptides of smaller size. It is possible that one of the cleaved SERT species is required for serotonin transport activity. Possible sites for specific proteolysis may be located in the region near TMD1 and in the intracellular loop between TMD4 and TMD5.

The state of the serotonin transporter protein in the platelets of patients with somatoform [correction of somatiform] disorders.

The role of the serotonin transporter protein (STP) in the development of somatoform [corrected] disorders was addressed in a correlational study of the levels of immunoreactive STP (IR-STP) using site-specific antibodies against the least conserved (among a group of other cotransporters) epitope at the C-terminal of STP and the level of anxiety symptoms in patients with somatoform [corrected] disorders. A total of 22 patients were studied, with DSM-IV diagnoses of somatoform [corrected] disorders, along with 32 mentally healthy subjects of comparable age and sex. Immunoblotting of IR-STP from patients from healthy donors produced a diffuse band between 68 and 105 kDal and a clear narrow band at 43 kDal. The 43-kDal IR-STP protein was almost completely absent from most patients, as compared with the levels of this protein in healthy donors. This result suggests an abnormality of STP processing or, perhaps, alternative splicing of the gene encoding STP in patients with somatoform [corrected] disorders, and this appears to reflect the dysfunction in serotoninergic transmission in the CNS in these patients.

[The status of serotonin protein -- a serotonin transporter in thrombocytes in patients with somatoform disorders]. / Sostoiane belka -- perenoschika serotonina v trombotsitakh u bol'nykh s somatoformnymi zabolevaniiami.

The role of serotonin transporter (SERT) protein in the development of somatoform disorders (SD) was investigated. An association study was performed in terms of the evaluation of the level of SERT immunoreactive (IR-SERT) protein using site-specific antibodies directed at SERT C-terminus fragment, poorly conserved among the other cotransporters. The level of the anxious symptomatology was also estimated in the patients with SD. 22 patients, who met DSM-IV criteria for somatoform disorders, and 32 normals were examined. In platelets from normals, IR-SERT protein migrated as a difuse band between 68 and 105 kDa, and a major sharper band at 43 kDa. Almost complete disappearance of platelet 43 kDa IR-SERT protein band was observed in most of the patients with SD. These findings permitted to suggest a possibility of either biosynthetic or processing abnormality of SERTs in the affected population, that might reflect a dysfunction of serotonin neurotransmission in CNS of the patients with SD.

Does platelet serotonin receptor supersensitivity accompany endogenous depression?

In a population of drug-free bipolar and unipolar depressed women, it was found that the concentration of serotonin sufficient to induce half-maximal shape change velocity in platelets is significantly (p less than 0.001) lower (0.13 +/- 0.0.04 microM) than that of closely matched controls (0.532 +/- 0.1 microM). This platelet concentration becomes higher after 1-3 months of antidepressant treatment (0.47 +/- 0.16 microM). Possible mechanisms for this up- or down-regulation of platelet serotonin receptor responsiveness are discussed.

Change in Bmax and Kd for [3H]flunitrazepam observed in the course washing rat brain tissue with distilled water.

A method for the removal of an endogenous inhibitor(s) of benzodiazepine receptor binding (extraction in distilled water at 20 degrees) was used to evaluate the possible influence of this inhibitor(s) on the affinity of the benzodiazepine receptor in vivo. The results indicate that the presence of an inhibitor(s) in the concentration which exists in the rat brain leads to a 30-fold increase in the Kd value as compared with the results obtained in vitro. The endogenous inhibitor(s) appeared to be a thermostable, proteinase K-resistant substance(s) with Mr between 500 and 10,000 daltons. Repeated washings of the brain tissue with distilled water were accompanied by a decrease in the Bmax for [3H]flunitrazepam. The [3H]flunitrazepam binding sites with Kd = 1.7 nM and Bmax = 61 fmoles/mg of protein were found in the supernatant collected after the second and third washings in distilled water. The presence of diazepam and the benzodiazepine antagonist Ro 15-1788 prevented in a dose-dependent manner the decrease in the Bmax value for [3H]flunitrazepam during the tissue washing with distilled water.

Increase of benzodiazepine binding to the membranes isolated in the presence of diazepam.

The preparation of brain cells in the presence of diazepam leads to a 60-70% increase in benzodiazepine receptor density. Diazepam may promote the dissociation of endogenous inhibitors (modulators) and/or retard the dissociation of benzodiazepin receptors from the membrane to the medium. Routinely used procedures for tissue preparation reveal only part of the benzodiazepine receptors originally present in the tissue.