Go Ark; an ameliorative bio-product (in vitro) on Phenyl induced cytotoxicity

Phenyl (PHY) is one of the chemicals which are used as a disinfectant in the world due to its toxic potential. Cleaning workers are directly exposed to it in institutes, hospitals and houses. Cow urine/Go Ark (GA) has been proved as a bioenhancer in many studies. The present study dealt with the in vitro analysis of PHY induced cytotoxicity (CT) on human peripheral blood lymphocytes and ameliorative potential of Distillate cow urine/Go Ark (DGA) and Fresh Go Ark (FGA) as GA is believed to be an elixir in Ayurved. MTT assay was used to study CT and Cell viability % on Human peripheral blood lymphocytes (HPBL) in vitro. CT of PHY was found to be higher than that of DGA and FGA treated groups. This showed that when PHY induced cells were treated with DGA and FGA, they showed increase in the cell viability %. It was also found that FGA had more potential for enhancing cell viability % of HPBL than that of DGA. We suggest that GA can be used as an ameliorative agent on PHY induced CT. It can be explored by in vivo experiments further for its detoxification properties. Now a day, PHY is used in combination with GA for cleaning purposes as “Gonyl”, it may be safe for cleaning workers to use GA based disinfectants to diminish the CT induced due to PHY exposure at the time of cleaning.

Cytokine production by peripheral blood lymphocytes in patients with ovarian epithelial tumors

In presence of autoserum, production of IL1b, IL2, IL4, IL6 and TNFa by peripheral blood lymphocytes of 30 patients with epithelial tumors of ovaries and 50 patients with epithelial ovarian cancer of I-IV stages was investigated. The data received enable to ascertain the increase of mononuclear activity during ovarian tumor progression, increase of T-helpers of the second type activity, along with the reduction of T-helpers of the first type activity in peripheral blood of patients, and abnormalities of IL6-dependent mechanism of control of ILIb and TNFa production by peripheral blood lymphocytes of patients with ovarian cancer.

IMMUNE CYTOPENIAS IN PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA (PECULIARITIES, PROGNOSTIC MARKERS)

Background: Immune cytopenia (IC) is one of the major complications in chronic lymphocytic leukemia (CLL). The paper describes the peculiarities of different immune cytopenia in CLL patients and the importance of individual prognostic markers in the course of the disease. Methods: We observed 62 patients with CLL complicated by immune cytopenia. Among these patients 30 had autoimmune hemolytic anemia (AIHA), 18 experienced immune thrombocytopenia (ITP), 10 had Fisher-Evans syndrome (FES), 3 were diagnosed with partial red cell aplasia (PRCA), and immune neutropenia (IN) was revealed in 1 patient. In addition to general examination and laboratory studies, the following examinations were performed: immunophenotyping of peripheral blood lymphocytes, flow cytometry (CD5; CD19; CD20; CD23; CD38; ZAP70), Coombs test, a molecular cytogenetic study of peripheral blood lymphocytes using the FISH method with TP53 and ATM probes, the level of ß2-microglobulin. Results: It was established that the overall survival of CLL patients with IC depends on the form of the latter. The median overall survival in patients with Fisher-Evans syndrome was the shortest (75 months), slightly better survival was observed in patients with AIHA (median 80 months), the best survival was found in patients with ITP (median not reached). Among unfavorable markers of CLL with IC, there is the presence of del 11q22.3. Unfavorable prognostic markers were also the following: a positive Coombs test, high levels of ZAP 70 expression, and high levels of ß2-microglobulin

Comparison and Evaluation of Different Radiotherapy Techniques Using Biodosimetry Based on Cytogenetics

While rapid technological advances in radiotherapy techniques have led to a more precise delivery of radiation dose and to a decreased risk of side effects, there is still a need to evaluate the efficacy of the new techniques estimating the biological dose and to investigate the radiobiological impact of the protracted radiotherapy treatment duration. The aim of this study is to compare, at a cytogenetic level, advanced radiotherapy techniques VMAT and IMRT with the conventional 3D-CRT, using biological dosimetry. A dicentric biodosimetry assay based on the frequency of dicentrics chromosomes scored in peripheral blood lymphocytes from prostate cancer patients and PC3 human prostate cancer cell line was used. For each patient blood sample and each subpopulation of the cultured cell line, three different irradiations were performed using the 3D-CRT, IMRT, and VMAT technique. The absorbed dose was estimated with the biodosimetry method based on the induced dicentric chromosomes. The results showed a statistically significant underestimation of the biological absorbed dose of ~6% for the IMRT and VMAT compared to 3D-CRT irradiations for peripheral blood lymphocytes, whereas IMRT and VMAT results were comparable without a statistically significant difference, although slightly lower values were observed for VMAT compared to IMRT irradiation. Similar results were obtained using the PC3 cell line. The observed biological dose underestimation could be associated with the relative decreased dose rate and increase irradiation time met in modulated techniques compared to the conventional 3D-CRT irradiations.

Changes in Glycanic Determinants of Lymphocytes Membranes in Peripheral Blood in Patients with B-Cell Chronic Lymphocytic Leukemia under Antitumor Therapy

The purpose of the study was to study the nature of changes in the exposure of surface glycans of peripheral blood lymphocytes in patients with B-cell chronic lymphocytic leukemia under conditions of antitumor therapy. Materials and methods. We studied the features of exposure of surface glycotopes of peripheral blood lymphocytes in patients with B-cell chronic lymphocytic leukemia under conditions of antitumor therapy using a set of seven lectins labeled with FITC and monoclonal antibodies to Tn-antigen- FITC for the detection of Tn antigen and CD43 exposure on blood lymphocytes. Cytostatic therapy included cyclophosphamide, vincristine (oncovin), prednisolone. Data were recorded on a Beckman Coulter EPICS flow cytometer. The results were processed using FCS3 Express. Results and discussion. The number of lymphocytes of healthy donors with a positive reaction to ConA, PHA-L, SNA, MAA-II and α1-acid glycoprotein amounted to 16.0±3.0%, 23.0±2.3%, 15.0±1.5%, 25.0±1.8% and 15.0±1.3%, respectively. The number of LABA-, UEA I-positive lymphocytes was 0.90±0.03% and 2.9±0.2%, respectively, and there was no binding to antibodies to Tn- and CD43-antigens. In the blood of patients with chronic lymphocytic leukemia, the level of ConA-, SNA- and MAA-II-positive lymphocytes increased relative to control by 2.2, 3.7 and 2.6 times, respectively. The number of LABA- and UEA I-positive lymphocytes in patients with chronic lymphocytic leukemia increased by 11 (p <0.01) and 23 (p <0.001) times and amounted to 10.5±0.5% and 67.5±5.5% respectively. The number of lymphocytes with CD43 antigen on their surface increased by 72 times, and the Tn antigen increased by 80 times. Cytostatic therapy reduced the level of LABA- and UEA I-positive lymphocytes by almost half, and MAA II-positive cells and lymphocytes interacting with antibodies to CD43 and Tn antigen by a third. The level of PHA-L-positive lymphocytes in the blood of chronic lymphocytic leukemia patients after undergoing alkylating therapy increased by 18.0±2.0% and almost did not differ from those obtained in the control group. Conclusion. 1. In chronic lymphocytic leukemia patients, the structure of glycoconjugates in peripheral blood lymphocytes changes, manifested in increased exposure of L-fucose, α-mannose and N-acetylneuraminic acid, which is confirmed by a significant increase in relation to the control of the number of ConA-, SNA-, MAA-II-, LABA I-positive cells. 2. Patients with chronic lymphocytic leukemia showed a significant increase in the number of lymphocytes, in which the markers of carcinogenesis CD43 and Tn antigens were found. 3. Cytostatic therapy significantly reduced the level of LABA-, UEA I- and MAA II-positive cells, as well as partially Tn- and CD43-antigen-positive lymphocytes, which indicates its positive effect on the treatment of chronic lymphocytic leukemia

Glycobiom Lymphocytes Surface Study of Patients with B-Cell Chronic Lymphocytic Leukemia

The purpose of the study was to investigate the intensity of exposure of peripheral blood lymphocyte surface glycans in patients with B-cell chronic lymphocytic leukemia by measuring the density of lectin- or antigen-positive epitopes under antitumor therapy in order to evaluate it for a more reasonable selection of qualitative and quantitative composition of therapy. Materials and methods. The objects of the study were blood lymphocytes of patients with chronic lymphocytic leukemia (n=15) aged 58-66 years before and after a course of standard chemotherapy according to the COP scheme. The control group consisted of healthy volunteers (n=15) aged 55 to 65 years. Isolation of lymphocytes was performed by a modified method of A. Boyum. Polyclonal antibodies to α1-acid glycoprotein and fibronectin were used. Exposure to Tn antigen and CD43 on blood lymphocytes was determined with secondary antibodies to mouse immunoglobulins conjugated to FITC (Millipore, USA). To study the exposure of glycans on the surface of lymphocytes, we used a set of seven lectins labeled with FITC. Data recording was performed on a Beckman Flower EPICS flow cytometer. Processing of the results was done using the program FCS3 Express. Results and discussion. Compared with the group of hematologically healthy donors on the surface of lymphocytes in patients with chronic lymphocytic leukemia, a 20-fold increase in the density of exposure to ConA epitopes, 10 times – UEAI- and SNA-positive epitopes were shown; MAA II epitope, Tn, and CD43 antigen densities were increased 100-fold (p <0.01). Exposure densities of MAA II-, Tn-, and CD43-positive epitopes on the plasma membrane of lymphocytes in patients with chronic lymphocytic leukemia receiving alkylation therapy decreased 10-fold relative to treatment data, but remained 10-fold higher than in the group of healthy hematologists. Conclusion. On the plasma membranes of lymphocytes in patients with chronic lymphocytic leukemia, the density of exposure of mannose and neuraminic acid residues was significantly increased. COP therapy reduced the density of these epitopes to control values. A significant increase in the density of carcinogenesis markers – Tn- and CD43-antigens on the plasma membranes of lymphocytes in patients with chronic lymphocytic leukemia has been shown. COP therapy provided only a partial decrease in their density, which indicates the insufficient effectiveness of COP therapy, its inability to completely stop the oncological process in patients with chronic lymphocytic leukemia

Genotoxicity of PM2.5 and PM1.0 Particulates on Human Peripheral Blood Lymphocytes in Manila, Philippines

Urban air quality is increasingly being studied as a fraction of the world’s population is living in megacities. In this study, particulate matter (PM) along Taft Avenue, Manila, the Philippines, is investigated in terms of its ability to induce genetic damage in human peripheral blood lymphocytes (PBLs). Size-segregated roadside air samples were obtained from 2015–2017 near a university gate and analyzed using in vitro micronucleus (MN) and cytokinesis-block proliferation tests. While cellular proliferation was unaffected by 0–0.1 kg/m3 of PM1.0 and PM2.5, PBL cells treated with PM2.5 displayed a significantly higher micronucleus count (p = 0.03) compared to the cells treated with PM1.0. Atomic absorption spectroscopy revealed greater amounts of Cd, Ca, Pb, K, Na, and Zn in PM2.5 compared to PM1.0. The results indicate that the differences in composition of the two size fractions of air particulates are associated with their genotoxicities.