Lymphocyte Blast Transformation Research Articles

INFLUENCE OF MODIFIED PEPTIDES FROM THE FETAL THYMUS ON THE ACTIVITY OF T-LYMPHOCYTES AND NATURAL KILLERS IN EXPERIMENTAL VIRAL HEPATITIS

The results showed that immunomodulin has a moderate regulatory effect on the spontaneous blast transformation of lymphocytes in the in vitro system. The costimulatory effect of immunomodulin in the reaction of PHA-induced blast transformation of T-lymphocytes was revealed. The data we obtained on the rather pronounced interferon-inducing properties of Sanogen developed by us and its combinations with well-known inductors -Cycloferon and Betaleukin, can be of practical use in the treatment of infectious pathology, especially viral hepatitis, when the combination of hepatoprotective, anti-inflammatory, antiviral, immunomodulatory and detoxification mechanisms of action, in the absence of toxicity and side effects, will ensure the development of sanogenetic processes in the patient's body.

May be dysfunction of cellular immunity considered a sign of post-COVID syndrome?

Our aim was to study association between proliferative activity of peripheral blood lymphocytes after COVID, and developing post-COVID syndrome, and to determine whether the cell immunity dysfunction may be regarded as its marker. The retrospective cohort study involved 242 patients (56 males, 186 females, 18 to 85 years old) who contracted new coronavirus infection. Of them, post-COVID syndrome was diagnosed in 180 cases (duration over 3 months). The patients were classified by severity of clinical course of COVID (i.e., presence of acute respiratory disease and pneumonias), and PHA-induced blast transformation of lymphocytes. Along with PHA-induced response, we studied cyclooxygenase (COG)-producing cells by morphological method. Control group consisted of 200 healthy people without any features of coronavirus infection. All patients were questioned and examined by multidisciplinary medical team, dependent on their complaints. We also registered incidence of comorbidities associated with cellular immune deficiency. The patients with post-COVID syndrome exhibited a decrease of PHA-induced lymphocyte proliferation as compared with control group (significant at p 0.01 in cases of acute respiratory infection, and p 0.05 in patients with pneumonia). Activity of COG-producing cells was similar in all groups, independently on presence of post-COVID syndrome. Classification of patients by presence of cellular immune dysfunction (PHA-induced blast transformation 50%) allowed to detect higher activity of COG-producing cells. This enzyme is known to participate in development of inflammation promoting immune deficiency, thus, probably, manifesting in clinical activation of herpesvirus infection following COVID-19. Activity of COG-synthesizing cells was found to be higher in post-COVID syndrome which evolves after middle-severe and severe forms of new coronavirus infection complicated by pneumonias. Chronic inflammation in post-COVID syndrome associated with high activity of COG-producing cells may promote dysfunction of cell immunity, thus being a cause of evolving syndrome, like as its biomarker. Absence of the immune cell dysfunction markers among other post-COVID features leads to decreased registration of post-COVID patients and misinterpretation of the results obtained.

The relationship between the character of the course of the disease, the level of tetanus and diphtheria antibodies, and indicators of general immunity in adults with infectious mononucleosis

ABSTRACT. EBV infection is one of the most common infections in the world. About 90 % of the world's population is infected with the Epstein-Barr virus. The Epstein-Barr virus is capable of affecting immunocompetent cells, causing lymphadenopathy, hepato- and splenomegaly, being stored forever in the body's lymphocytes, and causing an immunodeficient state. The aim of the study. The purpose of the study was to determine the relationships between the nature of the course of the disease for infectious mononucleosis (IM) in adults, the level of blood antibodies before tetanus and diphtheria, and general immunoreactivity. Materials and methods. In the first series of studies, blood antibodies was studied before diphtheria and tetanus in 79 adults. The control group consisted of 41 people. In the second series of studies, the immune status of 45 adults who became ill with IM was studied: 19 people who did not have a protective level of blood antibodies to diphtheria and tetanus (1st group) at the same time and 26 people (2nd group) who had a protective level of blood antibodies to these infections . In this work, the level of blood antibodies to diphtheria and tetanus toxin was studied by the ELISA method; population and subpopulation composition of blood lymphocytes using monoclonal antibodies: anti-CD3, CD4, CD8, CD22, CD16 (eBioscience, Beckman Coulter, R&D System). The proliferative activity of blood lymphocytes in the reaction of lymphocyte blast transformation was studied and the level of IgA, IgM, IgG in blood serum and the phagocytic activity of leukocytes. The results of the research were processed by the method of variation and correlation statistics using the "Statistica 10.0 for Windows" program. Results. It was established that 34.2% and 54,5 % of adults who developed MI in a moderate form did not have a protective level of blood antibodies for tetanus and diphtheria at the same time. A protective level of blood antibodies simultaneously before diphtheria and tetanus was observed in 55.2% of those who fell ill with IM in a moderate form and in 45,4 % of those who fell ill in a severe form. In persons who had a mild form of IM, the blood antibodies did not drop below the protective level before diphtheria and tetanus.The study of the immune status proved that in adults who do not have a protective level of blood antibodies against diphtheria and tetanus, the level of leukocytes, lymphocytes, CD3+, CD4+ - cells and the blast-transforming ability of T cells, the level of IgA, IgM, IgG, and the phagocytic ability of blood leukocytes are probably lower than in adults who have had an IM and have the protective blood antibodies level before these infections. Conclusions. The lack of a protective level of blood antibodies against diphtheria and tetanus in adults who have had IM is associated with a general low immunoreactivity of the body. A low level of blood antibodies before diphtheria and tetanus in persons who have had IM is closely related to the course of the disease. The lowest level of blood antibodies before these infections was observed in patients who fell ill with a severe form of IM, in patients with a mild form of the disease, significant changes in the level of blood antibodies before diphtheria and tetanus were not observed, as well as in the immune status.

Live attenuated vaccines in patients receiving immunosuppressive agents.

The use of live attenuated vaccines in patients with immunosuppressive agents is contraindicated in package inserts and guidelines in Japan and other countries. However, patients receiving immunosuppressants have a high risk of infectious disease becoming severe, and the necessity to prevent infectious disease is high. To date, 2,091 vaccinations have been reported in 25 reports of live attenuated vaccines in people receiving immunosuppressants. Twenty-three patients (1.1%) became infected with the virus strain used in the vaccine, which was varicella virus in 21 patients. No reports have described life-threatening complications. A prospective study at the National Center for Child Health and Development conducted under certain immunological conditions (CD4 cell count ≥ 500/mm3, stimulation index of lymphocyte blast transformation by phytohemagglutinin (PHA) ≥ 101.6, serum immunoglobulin G ≥ 300mg/dL) confirmed the serological effectiveness and safety. The evidence suggests that live attenuated vaccines can be used even in combination with immunosuppressants. Further evidence must be gathered and immunological criteria investigated to determine the conditions for safe use. Depending on the results of these investigations, the wording in package inserts and guidelines may need to be revised.

Influence of Paleobacteria on the Proliferative Activity of Human Lymphocytes In Vitro

When testing the proliferative activity of 14 strains of permafrost microorganisms in the reaction of blast transformation of human lymphocytes in vitro, a strain (Alcaligenes sp.) with mitogen properties was isolated (20-fold increase in the rate of lymphocyte proliferation in comparison with the control). Four strains activated lymphocyte proliferation by 3-9 times in comparison with the control. Three strains produced substances with cytostatic properties and reduced proliferation activity by 33-43% and one strain (Bacillus sp.) almost completely suppressed phytohemagglutinin-induced lymphocyte proliferation. These data indicate that strains with a unique immunobiological potential are concentrated in the population of permafrost microorganisms that have undergone rigorous evolutionary selection.

Is Formulary of Maranta Arundinacea Clarias Gariepinus (F-MaCg) a Potential Immunosti-mulant?

Background: External factors have the potential to act as immunostimulants in order to influence the body's protection from many foreign antigens. We intended to investigate the ethanol extract Formulary of F-MaCg effect as an immunostimulant. 
 Methods: A purely experimental with a completely randomized design was used on twenty-four white male rats. They were divided into four groups:1) G0 [given aquades (5 ml)]; 2) G1 [given F-MaCg-75 mg/gr BW (Body Weight)]; 3) G2 (F-MaCg -150 mg/gr plus Hepatitis B vaccine at the beginning and the end of treatment); and 4) G3 (F-MaCg -300 mg/gr BW plus hepatitis B vaccine at the end of treatment). The rat's spleen lymphocyte blast transformation was evaluated on the 15th and 37th days. Lymphocytes were examined using microtetrazolium assays. Optical Density (OD) was measured using an ELISA reader [493 nμ (nanomicro)]. Observation of lymphocyte viability by a counting chamber using a light microscope and trypan blue 1% before being cultured with Phytohaemoaglutinin.
 Results: Lymphocyte cell viability in the hepatitis B vaccine-induced group on the 15th day showed the highest average value in the G2 (1,484/mcl of blood); on the 37th day, it was in G3 (1,578/mcl of blood). The proliferative activity of spleen lymphocytes indicated by the difference in the OD values of the four treatment groups was 0.467, 0.913, 1.619, and 1.473 nμ, respectively. Histological observations of the spleen showed differences at all given formulary dose concentrations. 
 Conclusion: F-MaCg could be an immunostimulant because of its ability to trigger a cellular immune response.

The production of pro-inflammatory cytokines and the state of cellular immunity in patients with pulmonary tuberculosis with and without relapse

The purpose of the study was to conduct a comparative study of the synthesis of pro-inflammatory cytokines, the state of cellular immunity, free radical oxidation reactions and antioxidant protection in patients with pulmonary tuberculosis with and without relapse. Material and methods. The study was carried out on two groups of patients: the first group basic which included 39 patients with relapsed pulmonary tuberculosis (TBR); 2nd control group 39 patients with non-relapsed pulmonary tuberculosis (TB). Pro-inflammatory cytokines (TNF-α, IL-2 IL-6), CD-3 lymphocyte content, lymphocyte blast transformation response to phytohemagglutinin and tuberculin, NBT-test, AOPP, SOD, and catalase were determined in patients of both groups and healthy people. Results. In patients with tuberculosis with relapses, compared with patients without relapses, complaints of cough and dyspnea were more often presented, a longer duration of hospitalization, late abacillation. In patients with relapses, a pronounced suppression of the functional and specific activity of lymphocytes and neutrophils was noted, which is confirmed by high levels of lipid peroxidation (Advanced Oxidation Protein Products AORP) and low levels of the antioxidant system (SOD and catalase). The high content of pro-inflammatory cytokines in patients with relapses indicates a higher activity of the inflammatory process in these patients and corresponds to a low blast formation rate for both phytohemagglutinin and tuberculin. Conclusion. Сhanges in the content of pro-inflammatory cytokines, the state of cellular immunity and the characteristics of reactions of free radical oxidation and antioxidant protection indicate a higher activity of the inflammatory process in patients with pulmonary tuberculosis with relapses compared to patients without relapses, which led to a longer duration of hospitalization, late abacillation.

The effect of biostimulants from the spleen on the activity of T-lymphocytes and natural killers ..............

The work is devoted to the study of the effect of a biostimulator and its compounds with zinc on spontaneous, PHA-induced blast transformation of T-lymphocytes in patients with chronic viral hepatitis B, as well as on the cytotoxic activity of natural killers in colon cancer, and healthy donors in the in vitro system. The results showed that the biostimulator has a moderate regulatory effect on spontaneous blast transformation of lymphocytes in the in vitro system. The costimulating effect of the biostimulator of the PHA-induced blast transformation of T-lymphocytes was revealed. A significant increase in the membranotoxic function of natural killer cells in relation to K-562 target cells was found under the influence of a biostimulator after preliminary incubation of the drug with effector cells. The biological efficiency of splenic peptides increased after the addition of zinc ions to them.

Immune Disorders in Patients with Pulmonary Tuberculosis with Primary and Acquired Drug-Resistance of <i>Mycobacterium Tuberculosis</i>

The objective: to evaluate main parameters of cellular, humoral immunity and natural resistance in tuberculosis patients with primary and acquired multiple drug resistance; to study the correlation of these parameters with clinical and radiological manifestations.Subjects and Methods. A prospective study was conducted which included 169 patients with pulmonary tuberculosis, 80 of them were new cases of drug susceptible pulmonary tuberculosis and they made the Comparison Group (CG), and 89 patients suffered from multiple drug resistant tuberculosis and they made the Main Group (MG). The Main Group was divided into 2 subgroups: MG-1 – 40 patients with primary multiple drug resistant tuberculosis; MG-2 ‒ 49 patients who acquired drug resistance of Mycobacterium tuberculosis during treatment of tuberculosis.The immune assays included the lymphocyte blast transformation reaction (LBTR) to phytohemagglutinin and tuberculin (PPD), immunophenotyping of CD3+ and CD19+ cells; phagocytic index and phagocytic number were calculated; nitro blue tetrazolium test was used, concentrations of immunoglobulins and antimycobacterial antibodies were determined.Results. The clinical laboratory data demonstrated a more severe course of the disease in patients with multiple drug resistant tuberculosis, especially in those with acquired multiple drug resistant tuberculosis which was confirmed by the severity of immune deficiencies of the cellular immunity and innate resistance as well as by the overactivity of humoral immunity. The levels of CD3+ and LBTR reaction to PPD had a negative correlation with clinical manifestations and radiological features demonstrating the severity of pulmonary tuberculosis in patients with multiple drug resistance, especially in those with acquired multiple drug resistance; and CD19+ and antimycobacterial antibodies positively correlated with clinical manifestations and radiological signs.Conclusions. Parameters of cellular immunity and natural resistance were the lowest in the patients with acquired multiple drug resistance of Mycobacterium tuberculosis versus primary multiple drug resistance and drug susceptible Mycobacterium tuberculosis. Parameters of humoral immunity demonstrated the increased activity to compensate for the insufficiency of cellular immunity regardless of the type of resistance of Mycobacterium tuberculosis.

Investigation of Antiblastic and Antibacterial Properties of Binuclear Platinum (II)-Interferon Conjugate

Introduction: This work aimed to create a new platinum-interferon conjugate and investigate its properties. Background: Multiple drug resistance of tumor cells is a serious challenge in modern medicine. Neoplastic diseases are often complicated by persistent bacterial infections, which impairs efficacy and the outcome of anticancer therapy. The development of new bifunctional drugs for the treatment of tumor growth and bacterial complications is an urgent task. Objective: The objective of this study was to evaluate the complex antiblastic and antibacterial properties of interferon-platinum conjugate in vitro. Material and Methods: The coordination complex of dichloro-N,N,N,N-tetrakis-(2-aminoethyl)- 1,6-hexamethylenediaminobisplatin dichloride was used for production of conjugate with human α-interferon. The binding of a metal complex with protein was established by means of radiochemical neutron activation analysis. As test objects, we used lympholeukemic Jurkat cells line, human lymphocytes in reaction of lymphocyte blastic transformation and different strains of microorganisms, including Staphylococcus aureus and Pseudomonas aeruginosa. Results: Binuclear platinum human α-interferon conjugate has a significant dose-dependent antibacterial and antiproliferative suppressive effect on target cells and bacteria. Conclusion: The tested preparation can be of a certain interest in drug designing for rational antiblastic therapy, especially in the case of infectious complications. This conjugate can be a convenient platform for creating drugs for the complex therapy of oncological diseases with bacterial complications.

Comparison of modulation interference microscopy, DNA spectrometry, DNA cytometry, and flow cytofluorimetry in the assessment of phytohemagglutinin-induced activity of human blood lymphocytes

Rationale: The study of the structural particulars and functional state of immune cells and primarily lymphocytes is of great importance for both fundamental and clinical medicine. It requires the development of simple and reliable analytic methods that would allow for fast and effective real-time assessment of cell activity.Aim: To evaluate the effectiveness of the interference microscopy compared to DNA spectrometry, DNA cytometry, and flow cytometry with an internalized fluorescent label CFSE (carboxyfluorescein succinimidyl ester) in the assessment of PHA-induced proliferation of human blood lymphocytes.Materials and methods: Phytohemagglutinin (PHA)-induced proliferative activity of blood lymphocytes from 10 healthy volunteers was studied with various methodological strategies. Blast transformation of lymphocytes was induced by their incubation for 5 days with PHA 5 μg/mL. The cell proliferative activity was assessed as follows: 1) by DNA spectrometry at 260/280 nm using Tecan Infinite 200 Pro with a specialized NanoQuant Plate™; 2) by cytophotometry followed by cell distribution analysis assessing deoxyribonucleic acid (DNA) content after staining with Felgen's dye with an imaging system based on an Olympus BX41 light microscope with a ProgRes CF camera; 3) by flow cytometry using an internalized fluorescent label CFSE; the analysis was performed with a BD FACS Calibur flow cytometer; 4) by measurement of the lymphocyte interference profile with a modulation interference microscope MIM-340 (Schwabe, Russia). The functional activity of the nucleus (FAN) was determined and used as a criterion for assessment of the lymphocyte functional state.Results: Incubation of lymphocytes with PHA led to an increase in the linear size by 22.2±2.8%, a decrease in phase height by 46.3±4.7% (p=0.019), and an increase in FAN by 75.9±9.4%, vs control (p=0.046). As measured by isolated DNA spectroscopy, PHA stimulation of lymphocytes was associated with an increase in the amount of DNA by 55% vs baseline (409.8±22.3 ng/μL and 264.3±25.0 ng/μL, respectively, p=0.049). Felgen's reaction revealed that the proportion of nuclei containing more than 2n DNA was 2% in the control cells and 14.8% in the PHA-activated lymphocytes, with a difference between the groups of 12.8%. CFSE staining with subsequent incubation and assessment by flow cytofluorimetry demonstrated an increase in the percentage of proliferating cells from 1.68±0.9% in the control to 55.56±5.6% (p=0.00068) in the mitogen-stimulated sample.Conclusion: Modulation interference microscopy does not require the sample preparation and demonstrated comparable and even higher effectiveness compared to conventional methods for assessment of lymphocyte activity. At the same time, it allows for evaluation of the lymphocyte functional state in real time in the process of cultivation. This opens ample opportunities for evaluation immune cells for research and diagnostic purposes.

Cell-mediated immunity in patients with post-CoVID syndrome

Aim: to study the investigate of the cell-mediate immunity in post-COVID syndrome. Material and Methods: 278 patients with mild, moderate or severe Covid-19 in acute period with post-COVID syndrome were examined. All patients were questioned, and, if necessary, were examined by doctors of various specialties. The patients were divided into 2 groups depending on the severity of the course of the disease in the acute period and the duration of manifestations of post-COVID symptoms (up to 12 weeks or more). The state of cell-mediated immunity was assessed using the reaction of blast transformation of peripheral blood lymphocytes (morphological method for evaluating the reaction) in response to PHA. The activity of cells synthesizing cyclooxygenase was also determined. The control group included 200 practically healthy people of the same sex and age, who were not vaccinated and did not have a coronavirus infection. Results: a decrease in the proliferative activity of lymphocytes in response to PHA was found in patients who underwent Covid-19, both in mild (51,4±1.1%, p<0.01) and severe (49,3± 1,2% p<0,001), more pronounced in the latter in the first three months after the acute period of the disease (p<0,001). In all groups, there is an increase in the activity of cells synthesizing cyclooxygenase, which causes inflammation, which is more pronounced in patients who have had a severe form of Covid-19. Conclusion: The proliferative activity of peripheral blood lymphocytes in people who have had a coronavirus infection is reduced, both in the period up to 12 weeks and beyond; this is especially pronounced in patients who have had severe forms of Covid-19. In all groups, an increase in the activity of cells synthesizing cyclooxygenase, which plays a role in the development of inflammation, and possibly determines the progression of the course, was noted.

Comparative characteristics of the state of the immune system in patients with coronary artery disease with stable angina pectoris and acute coronary syndrome

The aim – to assess the relationship between the state of the immune system and the development of acute coronary syndrome in patients with IHD.Materials and methods. The first group consisted of 64 patients with ST-segment elevation acute coronary syndrome, mean age 54 (49–64) years; the second group – 223 patients with coronary artery disease with stable exertional angina, FC II–III, mean age 56 (49–63) years; the third group – 47 patients with acute coronary syndrome without ST segment elevation, mean age 61 (52–65) years. The material for the immunological study was peripheral venous blood. To determine the parameters of cellular and humoral innate and adaptive immunity in blood serum and supernatants of mononuclear cells, enzyme immunoassay was used.Results and discussion. In patients with coronary artery disease with acute coronary syndrome with ST segment elevation compared with patients with coronary artery disease with stable angina pectoris, the levels of indicators of the immune status in the blood were: CRP – 9.3 (5.3–12.0) versus 4.8 (2.4–8.1) mg/L (p=0.0001), sICAM – 785 (690–830) versus 565 (406–744) ng/ml (p=0.0001), IL-10 in blood mononuclear cells – 48 (1–228) versus 194 (21–758) pg/ml (p=0.0007), circulating immune complexes – 90 (70–108) versus 76 (54–105) od. (p=0.045), lymphocytes with apoptosis (CD95) – 16 (9–27) versus 11 (8–17) % (p=0.029), spontaneous oxygen-dependent metabolism of monocytes – 16 (12–21) versus 13 (9–17) (p=0.001). The levels of indicators of the immune system in the blood in patients with coronary artery disease with acute coronary syndrome with ST segment elevation compared with patients with coronary artery disease with acute coronary syndrome without ST segment elevation were: T-helpers – 37 (32–41) versus 42 (37–48) % (p=0.0006) (R=–0.33; p=0.0005), reaction of lymphocyte blast transformation to nonspecific antigen – 38 (32–47) versus 50 (42–61) % (p=0.0004) (R=–0.37; p=0.0003).Conclusions. The development of acute coronary syndrome is directly combined with increased activity of the immune system, as evidenced by the high production of proinflammatory CRP, IL-8, sICAM with a low level of anti-inflammatory IL-10, a pronounced humoral adaptive immune response (in terms of antibodies to the myocardium and vascular tissues, CD40, circulating immune complexes) and active functional state of monocytes (according to cNCT test, functional reserve, phagocytosis) in patients with coronary artery disease with acute coronary syndrome, regardless of the position of the ST segment in comparison with patients with stable coronary artery disease. Elevated levels of antibodies to the myocardium in patients with stable coronary heart disease indicate moderate myocardial damage due to temporary ischemia in angina attacks, even with a stable course of the disease. In patients with acute coronary syndrome, high levels of antibodies to the myocardium indicate myocardial damage due to increased ischemia in plaque destabilization much earlier than the clinical manifestations of acute coronary syndrome. In acute coronary syndrome with ST-segment elevation, compared with ACS patients without ST-segment elevation, activation of neutrophils and suppression of the activity of adaptive T-cell immunity is noted (by the level of T-helpers, sCD40L, blast transformation of lymphocytes, γ-interferon in mononuclear cells, apoptosis of lymphocytes).

Immunomodulatory Activity and Phytochemical Profile of Infusions from Cleavers Herb

Extracts from aerial parts of G. aparine (cleavers) constitute a herbal remedy with monography in British Herbal Pharmacopeia. On the European market, there are several drugs and food supplements consisting of Galium extracts. In folk medicine, cleavers was used topically in Europe, Asia, and the Americas to treat skin diseases. In several remedies, cleavers is also listed as an immunomodulatory active herb influencing the defense response of the human body. The aim of this study was to investigate the immunostimulatory activity and antioxidant potential in vitro of a raw infusion of cleavers and bioactive fractions. The functional activity of lymphocytes in the reaction of the lymphocyte blast transformation (RLBT) method was used for immunomodulatory activity assays and direct scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and hydrogen peroxide (H2O2) was chosen for the examination of antioxidant activity. It was shown that both the raw extract and fractions show significant immunostimulatory and scavenging activities. The obtained data partially justify the traditional use of cleavers as topical remedy for skin infections and for wounds.

Immunostimulating Activity of Gold-modified Nanodiamond Particles

Background: The fight against infectious diseases includes two main components – immediate direct anti-infective action and stimulation of one's own immunity. Objective: In this study, we investigated the properties of diamond nanoparticles modified with gold. The use of such gold nanoparticles as indirect anti-infectious agents and immunostimulators has certain prospects. Materials and Methods: Gold hydrosols were synthesized by the reduction in an aqueous solution of gold (III) with sodium citrate (Na3Cit) under heating. Modification procedure of nanodiamond by gold requires incubation, a small sample of nanodiamond in a defined volume of a gold sol for about 24 hours in a dark place. We use human blood cells as test objects. The reaction of blastic transformation of lymphocytes was applied here as a test of biological actions of modified nanodiamond. Results: Modified nanodiamond do not have a toxic influence on blood cells. Modified nanodiamond possesses stimulation effects on spontaneous proliferation of lymphocytes and does not significantly affect phytohemagglutinin-induced proliferation. Nanodiamond slightly increases phagocytosis parameters of neutrophil leucocytes. Conclusion: Thus, results showed that the nanodiamond modified by gold possesses immunostimulating activity, increases the phagocytic activity of neutrophilic leukocytes and stimulates lymphocytes in the spontaneous proliferation test. Gold-modified nanodiamond could be considered as a non-direct anti-infective agent through immune stimulation.

Phytochemical Profiles and In Vitro Immunomodulatory Activity of Ethanolic Extracts from Galium aparine L.

Galium aparine L., family Rubiaceae, is a widely spread species in the Galium genus. The herb of G. aparine is part of folk remedies and dietary supplements. In this study, we analyzed the chemical composition and immunomodulatory activities of G. aparine herb ethanolic extracts obtained from the plant material by maceration with 20%, 60% or 96% ethanol. The contents of hydroxycinnamic acid derivatives, flavonoids and polyphenols were determined spectrophotometrically, with extractives and polysaccharides quantified gravimetrically. The qualitative composition was studied using UHPLC-DAD-MS/MS analysis; isolation not previously described in G. aparine quercetin rhamnoglucoside was carried out through column chromatography, and the immunomodulatory activity of extracts was determined in the reaction of lymphocyte blast transformation. Major constitutes of extracts were iridoids, i.e., monotropein, 10-desacetylasperulosidic acid and asperulosidic acid; p-hydroxybenzoic acid; hydroxycinnamic acid derivatives, i.e., 3-O-caffeoylquinic, 5-O-caffeoylquinic, 3,4-O-dicaffeoylquinic, 3,5-O-dicaffeoylquinic, 4,5-O-dicaffeoylquinic acids and caffeic acid derivatives; flavonoids, i.e., rutin, quercetin 3-O-rhamnoglucoside-7-O-glucoside, and isorhamnetin 3-O-glucorhamnoside. Significantly, quercetin 3-O-rhamnoglucoside-7-O-glucoside was first isolated and identified in Galium species so far investigated. All G. aparine herb ethanolic extracts stimulate the transformational activity of immunocompetent blood cells, with 96% ethanolic extract being the most active. The data obtained necessitate further research into the mechanisms of immunomodulatory activity of extracts from G. aparine herb.

Assessing efficiency of synthetic peptide-containing spray in combination therapy of chronic generalized periodontitis

Antibacterial drugs are routinely used in therapy of periodontal diseases. However, increasing incidence of antibiotics resistance necessitates development of novel therapeutic approaches for oral diseases. Currently, newly designed antibacterial agents based on natural, semi-synthetic and synthetic peptides is the most promising approach in dentistry. Among them is a product containing synthetic peptide (ZP2) replicating active site of granulocyte-macrophage colonystimulating factor (GM-CSF) as the main active ingredient in Atsegram-spray (manufactured by Academic Innovation Research Center, Chelyabinsk). Our study was aimed at assessing efficacy of a combination therapy of inflammatory periodontal diseases by introducing Atsegram-spray as well as examining potential relationship between peptide ZP2-related biological properties, physicochemical properties of the spray and mechanism of antibacterial and immunotropic action for substantiating its application on oral mucosa. During the first stage experiments, it was found that the peptide ZP2 was able to trigger lymphocyte blast transformation in vitro indicating that it might influence cell proliferation and exhibit marked immunotropic activity. Next, we assessed potential effects of the peptide ZP2 on biofilm formation mediated by staphylococcal clinical isolates. It was shown that peptide ZP2 inhibited biofilm formation in 75.0±9.0% of S. aureus and 50.0±15.1% of S. epidermidis strains, with mean inhibition index of biofilm formation reaching 25.1±3.8 and 50.4±6.0%, respectively. However, peptide ZP2 in 8.3–25.0% of staphylococcal clinical isolates was found to stimulate/lack effect on biofilm formation by 14.9–48.5 and 16.7–25.0% cultures, respectively. Thus, the synthetic peptide ZP2 exerts divergent, but mainly inhibitory effects on biofilm formation with staphylococcal clinical strains, which are characterized by inter- and intraspecific (strain) variability. Use of a synthetic peptide-based spray in antibacterial therapy of mild chronic generalized periodontitis (main group) one month after the onset was found to improve oral hygiene by 28.5% as well as decrease PMA index and gum bleeding index by 82.8 and 100%, respectively. In contrast, such parameters in patients receiving basic therapy (comparison group) were lower on average by 2-fold. While analyzing physicochemical properties of the spray such as pH, buffer capacity and solution osmotic pressure, it was found that they were related to the antibacterial mechanism of drug activity and efficacy in treatment of inflammatory periodontal diseases. Thus, assessing peptide ZP2-related biological properties and physicochemical parameters of the spray allows to evaluate their role in mechanism of previously unknown antibacterial and immunotropic activity. These findings confirm feasibility and efficacy of using Atsegram-spray in dentistry as an alternative means to antimicrobial agents, such as antibacterial drugs.

Effectiveness of omega-3 fatty acid administration on completion rate of adjuvant chemotherapy for biliary tract cancer: study protocol for a single-centre, open-label, single-arm, historically controlled study

IntroductionMultimodal treatment prolongs the survival of patients with biliary tract cancer (BTC). However, the chemotherapy choices for this disease are few, and completing each chemotherapy session is important. Adjuvant chemotherapy...

Immunosuppressant Peptide Abu-TGIRIS-Abu-NH2 and its Application for Treatment of Multiple Sclerosis

Immunosuppressant peptide immunocortin for the first time was described in 1993. It corresponds to residues 11–20 of human Ig heavy chain (conserved motif of VH domain). There are no data about production of immunocortin by proteolysis of Ig in vivo. Synthetic immunocortin in concentration ~ 10−9 M suppresses phagocytosis in peritoneal macrophages, ConA-dependent blast transformation of rat lymphocytes, exhibits ACTH-like neurotropic activity and was suggested as a potential drug for treatment of a multiple sclerosis (MS). Here, we report a sequence and method of synthesis of Abu-TGIRIS-Abu-NH2 (Abu, alpha-aminobutyric acid), an artificial analogue of immunocortin. Biological trials of peritoneally injected Abu-TGIRIS-Abu-NH2 gave an evidence of its better efficacy versus immunocortin in a test for suppression of the experimental autoimmune encephalomyelitis (EAE) in Dark Agouti (DA) rats.

Реакція бластної трансформації лімфоцитів крові нелінійних самиць щурів, їх приплоду на ранніх етапах постембріонального розвитку на фоні впливу сольового екстракту Hirudoverbena

Реакція бластної трансформації лімфоцитів крові нелінійних самиць щурів, їх приплоду на ранніх етапах постембріонального розвитку на фоні впливу сольового екстракту Hirudoverbena