Background : assessment of the effectiveness of antipsychotic therapy is generally based on the knowledge, experience of the psychiatrist, and psychometric scales. However, all changes in a person’s mental state can be both a consequence and a cause of changes in biochemical processes, one of which is the complement system (CS). In addition to the fact that CS is the most important factor in innate immunity, its proteins, especially C3, are ubiquitous and directly involved in organismal and cellular homeostasis. CS includes its connection with coagulation, which compensates for the lack of activation potential of the cascade proteolytic complement system, as the first link of immunity. The deficiency is associated with excessive consumption of CS proteins in pathologically altered processes, which also determine mental pathologies. A change in the level of this connection allows one to determine changes in the overall level of homeostasis, for example, as a result of treatment. A new marker for assessing the level of coagulation-complement coupling, equal to the difference in the functional activity of CS in plasma and serum (d), may increase the reliability of assessing the effectiveness of therapy. Objective: justification for the use of this marker of the complement system in combination with mental state assessments on psychometric scales and electroencephalography results to analyze the effectiveness of therapy for patients with autism and schizophrenia aged 8–14 years. Patients and Methods : the study included 7 patients aged 8 to 14 years (10 ± 2.2) with a diagnosis “Childhood autism, F84.0 (ICD-10)” and 11 patients aged 11 to 14 years (13 ± 1.4) with a diagnosis “Schizophrenia, F20 (ICD-10)”. The assessment of the condition before and after treatment was carried out using the following scales: for patients with autism — CARS (Childhood Autism Rating Scale), BFCRS (Bush–Francis Catatonia Rating Scale), CGI-S (General Clinical Severity Scale); for patients with schizophrenia — PANSS (Positive and Negative Syndrome Scale) and CGI-S. A neurophysiological study of all patients was carried out with spectral analysis and topographic mapping using the NeuroKM computer system (Russia). A calculated parameter equal to the difference (d) of the functional activity of the complement system (faCS) in plasma and serum was used as a biological marker of CS. FaCS was assessed using the method on the ciliates Tetrahymena pyriformis and the Biolat device. Results: in 14 out of 18 patients, positive changes in mental state were observed, which corresponded to positive changes in marker d, as well as positive changes in ratings on scales and EEG results, in accordance with all assessments, 14 out of 18 patients from both groups showed positive changes in their condition, in 3 patients resistance to psychopharmacotherapy and minimal positive dynamics of the condition were noted; the marker of the complement system in these patients decreased slightly upon discharge; in one patient assessments of the biological marker and scales were contradictory. Conclusion: in most patients (14 out of 18 people), the selected therapy was effective, and the new candidate marker d of the complement system changed in the same direction (its normalization was noted) with clinical symptoms and neurophysiological parameters. The results of the study suggest that the candidate marker d reflects changes in the patients’ conditions as a result of treatment.

Relevance. Objective comparison of biological markers and real clinical presentation is especially difficult in mental disorders, which are classified according to a large number of diagnostic criteria and a wide variety of symptoms. Therefore, the development of an effective system of biochemical markers and assessment of their relationship to optimize the diagnosis and treatment of schizophrenia are relevant.The aim of the study was to develop a statistical model that combines known and tested biochemical markers for mental illnesses in patients with schizophrenia.Materials and methods. The study included 47 women aged 18–50 years (median age – 22 years) with the diagnosis of schizophrenia (ICD-10, F20) and 25 healthy women of the same age. The model was based on the functional activity of complement, thrombodynamics parameters, markers of inflammation, glutamate and energy metabolism, and antioxidant defense, which were shown to be associated with the severity of schizophrenia. The listed markers were evaluated in plasma, platelets, and erythrocytes of sick and healthy individuals.Results. Statistical software found pair correlations and features of the distribution of all markers as random variables in the examined groups and evaluated correlations between pairs of markers. Ten biomarkers were identified and united into a system that was adequately described by the logistic regression model. The model was evaluated using the Pearson’s test (χ2(11) = 57.6, p = 0.001) and calculation of correct predictions (91 and 80%) for samples of patients and healthy people, respectively.Conclusion. Calculating the logistic equation resulted in the probability that the patient has schizophrenia involving the immune system, hemostasis, and oxidative stress. This model can be considered as a new formalized approach to the preclinical diagnosis of mental illnesses.

Introduction The data of current research indicate the participation of systemic inflammation in the pathogenesis of endogenous psychoses. Changes in the level of peripheral immune markers are associated with the development of neuroinflammation and correlate with the severity of psychopathological symptoms detected in patients. However, the association between individual components of the immune system involved in the development of endogenous psychosis remains poorly understood. Objectives To study the connection between molecular and cellular components of the immune system in women with endogenous psychoses with depressive-delusional symptoms. Methods 32 female patients aged 23 [17; 36] years with endogenous psychoses within different nosologies (F20, F21, F31, depressive-delusional conditions) and 17 women without clinical signs of psychiatric pathology were examined. The activity of leukocyte elastase (LE), α1-proteinase inhibitor (α1-PI), the proportion of four subpopulations of monocytes (classical CD14++CD16-, intermediate CD14++CD16+, nonclassical CD14+CD16+ and transitional CD14+CD16-) in plasma, activity of cytochrome-c oxidase (COX), glutamate dehydrogenase (GDH), glutathione s-transferase (GST) and glutathione reductase (GR) in platelets and functional activity of complement system (faCS) in serum were determined. The PANSS scale was used to assess the severity of psychopathological symptoms. Results Increased activity of the inflammatory markers LE (p=0.033) and α1-PI (p=0.02) was found in the plasma of the patients. Increased percentage of pro-inflammatory monocytes (intermediate and transient) in plasma (p=0.003) was confirmed by negative correlations between CD14++CD16- and CD14++CD16+ (R=-0.685, p=0.00002), CD14++CD16- and CD14+CD16- (R=-0.608, p=0.0002), CD14++CD16- and CD14+CD16+ (R=-0.424, p=0.002). A decrease in GDH activity (p=0.0079), GST activity (p=0.002) and GR activity (p=0.0006) was observed in patient platelets, which can reflect changes in the activity of intracellular metabolic pathways. A positive correlation was found between COX activity and α1-PI (R=0.51, p=0.025). A significant decrease in faCS compared to control (p=0.0003) and a negative correlation between faCS and GST activity (R=-0.496, p=0.011) were observed. faCS was positively correlated with the degree of reduction in the PANSS score (R=0.416, p=0.038). Conclusions The revealed connection between molecular and cellular components of the immune system in patients with endogenous psychoses reflect activation of the systemic inflammatory response accompanied by changes in the ratio of monocyte subpopulations and impaired regulation of the complement system. The data obtained can be used to develop methods of monitoring patients taking into account their immunological features. Disclosure of Interest None Declared

The effects of extracts of ten plant species from Russia and five species from Vietnam on the growth and survival of ciliates Tetrahymena pyriformis were studied. T. pyriformis belongs to the subkingdom Protozoa, which also includes pathogens of protozoan infections. Extraction of dried plants was carried out with acidic and alkaline aqueous solutions, as well as with an aqueous ethanol. Various amounts of extracts were added to the ciliate cells, and the number of cells survived after incubation for 1 and 24 h was recorded. We found that our samples of several plants, including wormwood, harmala, and licorice, similarly to those studied earlier, exhibit antiprotozoal activity, which may indicate that the secondary metabolites are the same in plants from different regions. Using the ciliate T. pyriformis as a model organism, the presence of antiprotozoal activity in extracts of lilac, chondrilla, cinquefoil, hop, and elm was shown for the first time.

As human civilization develops, environmental problems arise more and more often. All forms of the environment are consistently exposed to pollution: water, air, and soil. In the natural aquatic environment, redox processes occur, in which both endogenous and exogenous substances are involved. The processes of antagonism and synergy are also realized, which leads to a decrease or increase in the toxicity of the environment. All this affects intra-aquatic processes, which, as a result, lead to the formation of the quality of the aquatic environment. Historically, there have been three main approaches to assessing water quality using biological objects. These are biodiagnostic methods: bioindication, bio marking, and biotesting. The review reveals the features of biodiagnostic methods and their application in assessing the quality of natural waters.

To search for compounds with antiprotozoal activity, effects of snake venoms on the ciliates Tetrahymena pyriformis was studied. T. pyriformis from subkingdom of Protozoa, including the protozoal pathogens, was used as a model organism to select the venoms that are the most active against parasitic protozoans. Various concentrations of venoms were added to the cells, and the cells that survived after 24 h were counted. Among the six snake species from the Viperidae family, the venom of the viper Vipera berus, which completely killed the cells at 49 μg/mL, was the most active. Among four species from the Elapidae family, the previously studied cobra venoms containing cytotoxins with strong antiprotozoal activity as well as the venom of krait Bungarus multicinctus (10 μg/mL) were the most active. The venoms of the pit vipers and Nikolsky's viper did not show any activity at 12.5 mg/mL. Thus, the venoms of V. berus and B. multicinctus are promising for the isolation of new antiprotozoal compounds.

The cytotoxicity of doxorubicin (Dox) and its peptide modifications Z-Gly-Pro-Dox and Boc-Gly-Pro-Dox were studied. Tetrahymena pyriformis was used as a test system, which made it possible, due to the short life cycle and high reproduction rate of ciliates, to trace their response to the effects of toxicants over several generations. It was found that peptide modification of the Dox molecule markedly reduces its cytotoxic and cytostatic effect. The Z-Gly-Pro-Dox modification has less cytotoxic and cytostatic effect compared to Boc-Gly-Pro-Dox. When determining the ability of drugs (at a concentration of 100 μM) to prevent bacterial contamination of samples, it was shown that the smallest degree of overgrowth was recorded in the presence of Dox (OD600nm 81.1). Boc-Gly-Pro-Dox also had a bacteriostatic effect, though less pronounced (OD600nm 93.8). The degree of overgrowth in the presence of Z-Gly-Pro-Dox was close to that of distilled water. The results obtained on ciliates did not contradict the data obtained in similar studies on mice.

Introduction: the complement system can be a critical factor in the outcome of SARS-CoV-2 viral infection. Many mental illnesses are characterized by systemic sterile inflammation, in which the complement system is an obligatory participant. Purpose: to present an analysis of scientific data on the role of the complement system in the pathogenesis of viral diseases and the characteristics of the course of COVID-19 in mental patients. Material and methods: the keywords “complement system” “SARS-CoV-2”, “inhibition of the complement system”, “COVID-19” “mental illness” were used to search scientific articles in the databases MEDLINE, PubMed and other bibliographic sources. Conclusion: patients with mental illness are at risk due to physiological and mental characteristics, and infection with SARS-CoV-2 can provoke a relapse of the underlying disease. Therapeutic inhibition of complement system will help reduce this risk and reduce the likelihood of severe complications from systemic inflammation caused by this infection.

How a number of biological effects of the peptide neuroprotector HLDF-6-amide work are still unknown and require further study. It was found that HLDF-6-amide dose-dependent (1–100 μM) stimulates the proliferation of Tetrahymena pyriformis infusoria, and the MK801 glutamate NMDA receptor antagonist partially blocks this effect. The introduction of a peptide (12.5–100 μM) reduces the cytotoxic and cytostatic effect of doxorubicin (25 μM) on infusoria. Radio-ligand analysis revealed the presence of specific binding sites of [3H] Ac-HLDF-6-amide on the cell membranes of the rat brain cortex, which indicates the receptor mechanism of action of the peptide. Thus, it was shown for the first time that HLDF-6-amide stimulates proliferation and has cytoprotective properties, which may underlie its wound-healing, nootropic and neuroprotective effects. The participation of the glutamate system, in particular ionotropic NMDA receptors, in these effects was established.

It is known that functional activity of complement system depends not only on balance and concentration of components participating in formation of the system end products, but also on levels of inhibitory activities. Numerous relations with hemostasis also substantially contribute to general level of complement system activity. Changes in complement system functioning are inevitable during chronic diseases accompanied with immune system dysregulation. All mental diseases tend to be chronic and are they aggravated by patients’ immune system changes. Autism spectrum disorders in children is a group of mental disorders. Immune system dysregulation is usually detected in such patients, manifesting as excessive susceptibility to viral and bacterial infections. Therefore, the level of its functional activity is diagnostically and prognostically significant in this pathology, since the complement system is a key element of immune system.We have evaluated functional activity of complement system in patients with autistic spectrum disorders, using the method which was developed earlier. It is based on the reaction of the protozoa (Tetrahymena pyriformis) which are both targets and activators for the complement system. The complement system capacity (cSC) was used as the main parameter of complement evaluation. The half-time of protozoa survival (T50) was defined using the BioLat device for each serum specimen added at four concentrations (1/20, 1/40, 1/80, 1/160 dilution). The complement capacity was calculated as the area enclosed by influence curve of the reciprocals of T50 and the serum dilution. According to Mann–Whitney U test, the difference between patients’ and healthy volunteers’ groups was established as Z = 4.43 (by T50 at 1/160 dilution), p < 0.001 and by cSCas Z = 5.8, p < 0.001. cSC was calculated from the results obtained at each serum concentration measured. The difference between the two groups according to Mann–Whitney U test appeared to be more significant than the difference according to T50. Therefore, cSC was taken as the main characteristic of complement system function.The contribution of hemostasis plasma components to complement system functional activity level was estimated by determination of complement capacity in plasma and serum of each blood sample from 6 patients with autism spectrum disorders and 5 healthy donors. All healthy donors showed small difference between plasma and serum complement capacity, and their complement activity was higher in plasma. In patients’ group, the complement capacity levels in plasma and serum differed significantly. The cSC levels of two patients were higher in serum than in plasma, and the cSC levels of three other patients were significantly higher in plasma than in serum. Differential involvement of coagulation into the complement system activation may be indicative for the immune system dysfunction which is observed in patients with autistic spectrum disorders of different etiology.

It is known that psychotropic substances affect the immune system. Unfortunately, chronic antipsychotic administration causes side toxicological effects, associated with oxidative stress. The mechanisms of these effects are still unclear. We investigated the impact of sub-chronic administration of haloperidol (Hal) on parameters of innate immunity and related systems in healthy rats and compared them with Hal content. Hal administration (0.5 mg/kg, 3 weeks) resulted in two-fold decrease of the activity of the complement system and hemostasis. Hal content correlated with the activity of the complement (r = −0.71), phagocytic activity of peritoneal macrophages (r = 0.78), leukocyte elastase (r = −0.71) and glutathione-S-transferase activity (r = −0.67). Hal fully blocked in vitro PMA-induced iNOS expression in macrophages and changed their morphology to “anti-inflammatory” phenotype. The comparison of in vivo and in vitro data showed that Hal has a direct effect on phagocytic component of innate immunity and an indirect effect on leukocyte elastase and antioxidant enzymes. The results obtained in the present study indicated that Hal significantly affects homeostasis and causes a number of complex biological transformations.

Bacterial cellulose is a biomaterial produced by Gluconacetobacter hansenii bacteria with peculiar properties. The aim of this research was to assess three methods for washing bacterial cellulose from producer cells: 5% SDS, RIPA and RIPA with the addition of nucleases. Atomic force microscopy showed that all methods provide the removal of bacterial cells. The authors carried out a quantitative estimation of residual DNA on the bacterial cellulose matrix and demonstrated a significant difference (p < 0.05) in the DNA content between the group of RIPA with the addition of nucleases and the non-washed matrix. The authors evaluated the cytotoxicity of the extracts and the matrix of bacterial cellulose in regard to protozoa (Tetrahymena) and the mammalian cell culture (NIH/3T3), respectively. The results from the assay with the Biolat device and the MTT assay indicated that the samples are not cytotoxic. The Live/Dead assay of NIH/3T3 showed that bacterial cellulose is an adhesive matrix in regard to mammalian cells and maintains cell viability. In accordance with the authors' data, a bacterial cellulose matrix may provide with washing 5% SDS and RIPA; however, the addition of nucleases is necessary to reduce residual bacterial DNA.

Introduction: hemostasis is one of the most important protective systems of the organism, and its deviations from the norm can lead to fatal results. Changes in hemostasis are observed in many diseases, including mental pathology. In addition, parameters of this system may also change during antipsychotic therapy. Objective: Development and testing of a new method and device for estimating plasma hemostasis parameters in patients with mental pathologies in order to optimize treatment and eliminate fatal outcomes. Materials and methods: in the study we used frozen (-80 °C) platelet-free blood plasma (PFP) samples from patients with schizophrenia (16 patients), taken at admission to the clinic FSBSI «Mental Health Research Centre». Plasma coagulation of patients and healthy donors was assessed with the device of our design. Hemostasis assessment is based on the determination of the clot size in proportion to the brightness of the image, which changes as a result of clot formation and lysis. Results: optimal concentrations of coagulation and fibrinolysis activators, as well as concentrations of calcium ions were determined during the first stage of testing the method and device. The median values of all parameters of coagulation and fibrinolysis except «time before the formation of a clot» in patients upon admission to the clinic were reduced relative with combined plasma donors. The rates of clot formation and lysis in patients are significantly reduced at discharge compared with these parameters at admission. Consequently, these parameters can serve as an estimate of the change in the state of the coagulation and fibrinolysis systems during treatment. Conclusion: during approbation of the device and method in the process of coagulation and fibrinolysis research in patients with schizophrenia, lower levels of parameters were detected compared to donors, and a decrease in the activity of plasma coagulation and fibrinolysis during treatment with antipsychotics was shown.

Toxicity of human blood serum for ciliate Tetrahymena pyriformis is determined by the complement system. When ciliate are dying after being exposed to blood serum, cell membrane permeability for low-molecular-weight compounds significantly increases, probably due to pore formation. Serine protease inhibitors or exposure to physical factors inducing complement inactivation (e.g., heating up to 56°C) completely prevented ciliate death under the effect of human serum. Activation of serum complement upon interaction with Tetrahymena cells occurred by the classical or lectin pathway, while the contribution of the alternative activation pathway was negligible.

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.

Complement system is an important component of innate immunity, providing primary protection against pathogens invading the body. In addition, it was shown that the complement system is associated with many diseases, not only autoimmune and infectious, but also mental disorders. In this regard, it is necessary to develop affordable and fast method of measuring activity of the complement system in real-time mode. We present a new semi-automated method for assessment of serum complement activity. The assay is based on cytolytic action of complement system upon the ciliate organism Tetrahymena pyriformis. This method consists in repeated counting of live Tetrahymena motile cells by means of specially developed Biolat device, which consists of two video cameras, light sources, and movable round plate. The plate has two rows of holes. The device also includes microprocessor control unit based on AutoCiliata software, intended for control of operation module and counting the surviving cell. The calculations are based on fixation of two sequential video-frames, with subsequent software image processing. Cell death events were observed upon incubation in triethanolamine (TEA) buffer containing 5% of blood serum. We have also compared complement activity in different buffers, i.e., standard medium for culturing of ciliates, Veronal-Medinalum buffer, and the TEA buffer. TEA buffer was found superior to the Veronal buffer when applied in the test system. The time of cell death in the TEA-buffered medium containing 5% serum was < 15 minutes for all the sera studied. The parameters denoting serum complement activity were as follows: a half-life time for the moving cells (TLD50), and a similar value for 100% cell inactivation (1/TLD50, functional activity of the complement system, ACS). The sensitivity of this assay was calculated from dependencies between TLD50 and ACS, and actual serum concentrations. We have suggested an opportunity for evaluation of an integral complement activity, and interrelations between the intensity of synthesis and consumption of its major effector proteins. In the course of this study, we have tested different concentrations of Ca++ and Mg++ ions in the incubation buffer, with optimal physiological concentrations of2.5 mMand1.5 mM, respectively. We have also estimated statistical precision characteristics for pre-analytical and analytical steps of the method. The average coefficients of variation (CV) were 3.9% and 2.7%, respectively, thus satisfying the reliability criteria in research. A short performance time of the study suggests its potential application in clinical practice, including online examination regimens. A method for semi-automatic measurement of serum complement activity could be applicable in daily clinical practice, including the online performance.

A new universal method for measuring activity of the serum complement system in humans, laboratory, domestic, agricultural animals, birds and amphibians is based on automated evaluation of the mortality of ciliate Tetrahymena pyriformis under the effect of the complement system. In contrast to the hemolytic method, measured activity of the complement shows no erroneously high results caused by reactive lysis in febrile patients. The method can be used for studies of the complement system in humans and animals without species-specific adaptation.