scholarly journals Monitoring of cytogenetic instability by micronuclei assay of both immunocompetent and non-immunocompetent cells in tick-borne encephalitis patients depending on variants of glutathione-S-transferase genes in the genotype

2019 ◽  
Vol 9 (3-4) ◽  
pp. 600-606
Author(s):  
Nikolay Nikolaevich Ilyinskikh ◽  
Ekaterina Nikolaevna Ilyinskikh ◽  
Evgenia Vladimirovna Zamyatina ◽  
Svetoslava Vyacheslavovna Lee

Aim of this study was to study the dynamics of the frequency of cytokinesis-blocked T-lymphocytes with micronuclei in peripheral blood and the frequency of buccal micronucleated epithelium cells for a period of half a year in patients with acute tick-borne encephalitis, depending on burden of active and inactive variants of glutathione-S-transferase genes (GSTM1 and GSTT1) in the patient's genotype. We carried out micronucleus assay in immunocompetent and non-immunocompetent cells in 54 patients with acute tick-borne encephalitis and 35 healthy persons (control) residing in the Tomsk and Tyumen regions. To analyze the frequency of cytokinesis-blocked micronucleated T-lymphocytes was used venous peripheral blood as material for phytohemagglutinin-stimulated cultures, and to study the frequency of buccal micronucleated cells, samples of the buccal mucous membrane epithelial cells were obtained. To carried out both techniques of micronucleus assay, cytological preparations were prepared, which were stained using the Giemsa or Felgen methods. The material for the study was obtained repeatedly during admission of patients to treatment, and also after 1 week, 1, 3 and 6 months.  Polymerase chain reaction was used to analyze the alleles of the GSTM1 and GSTT1 genes. As a result of this analysis was found a significant increase in the frequency of micronucleated cells in tick-borne encephalitis patients compared with the control group. In addition, the frequency of cytokinesis-blocked micronucleated T-lymphocytes was increased significantly higher than the one of micronucleated buccal cells. The most significant and prolonged increase in the frequency of micronucleated cells was associated with the mutant inactive variants of the genes GSTM1 (0/0) and GSTT1 (0/0). In the patients with burden the inactive forms of these genes, the cytogenetic instability of the cytokinesis-blocked blood T-lymphocytes could persist for up to six months. In case of buccal cells, the frequency of micronucleated cells was close to the one in the control group as early as 1-3 months after a course of treatment. Conclusion. It was found that the most increased and prolonged frequency of cytogenetically instable cells persisted in cytokinesis-blocked T-lymphocytes of peripheral blood of patients with tick-borne encephalitis who were carriers of the genotype with inactive variants of  both GSTM1 (0/0) and GSTT1 (0/0 ) glutathione-S-transferase genes.

2021 ◽  
Vol 19 (1) ◽  
pp. 01-05
Author(s):  
K Sriambika ◽  

Background: Formaldehyde (FA) is the reactive and simplest of all the aldehydes. It is used as a preservative in anatomy, pathology and forensic laboratories. The international agency for research on cancer has classified FA as a carcinogen that can cause nasopharyngeal carcinoma, Leukaemia, Liver and pancreatic cancer. Objective And Method: The aim of the study was to assess the DNA damage in peripheral blood lymphocytes and in buccal cells by Micronucleus assay in Formalin exposed workers of Anatomy, Pathology and Forensic laboratories and compare with the control group, and also to analyze the relationship between frequency of Micronuclei and duration of exposure to formalin. Results: The mean and standard deviation (SD) of micronuclei in peripheral blood of exposed was 8.35 and in controls was 4.18. There was a significant increase in the frequency of MN in exposed group when compared with the comparison group (p<0. 5876). Pearson’s correlation test showed a positive correlation between the years of FA exposure and the number of micronuclei in buccal cells and peripheral blood indicating that DNA damage due to FA was directly proportional to the duration of exposure (r=0.8, 0.9). Conclusion: The present study was done to assess the DNA damage in people who were exposed to FA and a control group not exposed to FA by buccal cell and peripheral blood Micronucleus Assay. There was a significant increase in the MN in people exposed to FA which was directly proportional to the duration of exposure.


2019 ◽  
Vol 22 (3) ◽  
pp. 225-232
Author(s):  
Nikolay N. Ilyinskikh ◽  
Marina S. Kostromeeva ◽  
Ekaterina N. Ilyinskikh

BACKGROUND: Tick-borne encephalitis (TBE) is an acute viral disease with activation of oxidative stress and increasing in cytogenetic instability. Clinical symptoms of infectious diseases usually more severe in patients with type 2 diabetes mellitus (DM2), especially in the case of burden in the genotype of mutant variants of glutathione-S-transferase genes GSTM1 and GSTT1. AIMS: The aim of this study was to study the dynamics of the frequency of micronucleated cells in patients with acute TBE with concomitant DM2, depending on the burden of active and inactive variants of glutathione-S-transferase genes (GSTM1 and GSTT1) in the patients genotype. MATERIALS AND METHODS: Totally, samples to make micronucleus assay were obtained from 138 patients with febrile illness of acute TBE, 64 of whom were diagnosed with concomitant DM2 (groups 3 and 4). As control groups, 57 healthy individuals (control 1) and 61 patients with DM2 (control 2) were examined. The samples of buccal cells for the micronucleus assay were repeatedly obtained from the individuals on the first day of admission, and also after 1 week, 1, 3 and 6 months. Polymerase chain reaction was used to analyze the variants of the GSTM1 and GSTT1 genes. RESULTS: On the first days of the disease, significant increases in the frequency of micronucleated buccal cells were determined in all TBE patients as compared to controls 1 and 2 (P0.001). Significant increases in the frequency of micronucleated buccal cells was revealed in groups 3 and 4 of the TBE patients who were carriers of inactive variants of the GSTM1(0) and GSTT1(0) genes, as compared to the subgroup of TBE patients with active variants of these genes (P0.001). In all subgroups of TBE patients with concomitant DM2, the frequencies of micronucleated cells were significantly higher than in the subgroups of TBE patients without DM2 (P0.001). Study of the dynamics of the frequency of micronucleated buccal cells, as compared to the control, demonstrated that the highest and long-lasting (within 6 months) cytogenetical effects were maintained in the group of TBE patients with genotype GSTM1(0)/GSTT1 (0) and concomitant DM2. CONCLUSION: The most prolonged and highest increases in the frequencies of cytogenetically instable cells were determined in the group of acute TBE patients with concomitant DM2 who were carriers of the genotype with inactive variants of both GSTM1(0) and GSTT1(0) glutathione-S-transferase genes.


2017 ◽  
Vol 14 (1) ◽  
pp. 24-29
Author(s):  
Ekaterina N. Ilyinskikh ◽  
Nicolai Nicolaevich Ilyinskikh

Backgraund. Tick-borne encephalitis (TBE), one of the large-scale public health problems of the Siberian region, can cause significant cytogenetic damage in humans by stimulating oxidative stress. Active functioning of the detoxification system including enzymes of glutathione-S-transferase is intended to protect the genetic structure of cells in the body, while at the same time, there are studies showing the defective functioning of this system in individuals with obesity. Aims. Aim of this work is to study the longevity of term of TBE-induced cytogenetic damage in patients with different alleles of genes of glutathione-S-transferase, suffering from alimentary-constitutional obesity compared with the control. Materials and methods. We were examined 133 female residents of the northern areas of Tomsk region from 35 to 44 years old who were hospitalized for infection TBE at local medical clinics. All individials were divided into two subgroups. Patients of the 1st subgroup (control) had a body mass index (BMI) within 2126, the 2nd subgroup had BMI showing severe obesity (BMI 3545). Buccal cells for cytogenetic and molecular analysis were taken several times: 12 days after hospitalization, 1 week, 1 month, 3 months, and 6 months after beginning of the disease. Methods of immunoassay and polymerase chain reaction (PCR) was used to confirm the diagnosis of TBE. As an untreated control were examined 126 healthy female residents of the same age groups. All individuals signed informed consent. In the analysis of deletions in the genes GSTM1 and GSTT1 was used multiplex PCR. We studied at least 1000 epithelial buccal cells from each individual. Results. In patients with high BMI rates revealed a significant increase in the frequency of the buccal cells with cytogenetic damage, compared to the control group. TBE disease was significantly increased the frequency of the cytogenetically damaged cells in this group of patients. The cytogenetic aberrations persisted in their body for six months after hospitalization. In patients with normal BMI rates TBE induced significantly less changes. The recovery of normal cytogenetic status was observed in this group 3 months after hospitalization. Analysis of the glutathione-S-transferase gene variants showed that in TBE patients with normal BMI and non-active alleles of these genesthe frequency of micronucleated epithelial cellswere significantly increased compared to the patients with the activeGSTM1 (+) / GSTT1 (+)genes. In the patients with alimentary obesity such patterns were not observed. Conclusions. Thus,infection with TBE in patients with obesity induced significant increase in both the frequency of cytogenetically damaged cells and the longevity of their persistency in the body compared to these in control group. Polymorphisms in genes of glutathione-S-transferase had no significant effect on the indices.


2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Yixuan Liu ◽  
Suhong Xie ◽  
Lei Li ◽  
Yanhui Si ◽  
Weiwei Zhang ◽  
...  

Abstract Background This study investigates the effect of autologous bone marrow transfusion (BMT) on the reconstruction of both bone marrow and the immune system in patients with AIDS-related lymphoma (ARL). Methods A total of 32 patients with ARL participated in this study. Among them, 16 participants were treated with conventional surgery and chemotherapy (control group) and the remaining 16 patients were treated with chemotherapy followed by autologous bone marrow transfusion via a mesenteric vein (8 patients, ABM-MVI group) or a peripheral vein (8 patients, ABM-PI group). Subsequently, peripheral blood and lymphocyte data subsets were detected and documented in all patients. Results Before chemotherapy, no significant difference in indicators was observed between three groups of ARL patients. Unexpectedly, 2 weeks after the end of 6 courses of chemotherapy, the ABM-MVI group, and the ABM-PI group yielded an increased level of CD8+T lymphocytes, white blood cells (WBC), and platelet (PLT) in peripheral blood in comparison to the control group. Notably, the number of CD4+T lymphocytes in the ABM-PI group was significantly higher than that in the other two groups. Additionally, no significant difference in haemoglobin levels was observed before and after chemotherapy in both the ABM-MVI and ABM-PI groups, while haemoglobin levels in the control group decreased significantly following chemotherapy. Conclusions Autologous bone marrow transfusion after chemotherapy can promote the reconstruction of both bone marrow and the immune system. There was no significant difference in bone marrow recovery and reconstruction between the mesenteric vein transfusion group and the peripheral vein transfusion group.


2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Márcia Fernanda Correia Jardim Paz ◽  
André Luiz Pinho Sobral ◽  
Jaqueline Nascimento Picada ◽  
Ivana Grivicich ◽  
Antonio Luiz Gomes Júnior ◽  
...  

This study aimed to evaluate DNA damage in patients with breast cancer before treatment (background) and after chemotherapy (QT) and radiotherapy (RT) treatment using the Comet assay in peripheral blood and the micronucleus test in buccal cells. We also evaluated repair of DNA damage after the end of RT, as well as the response of patient’s cells before treatment with an oxidizing agent (H2O2; challenge assay). Fifty women with a mammographic diagnosis negative for cancer (control group) and 100 women with a diagnosis of breast cancer (followed up during the treatment) were involved in this study. The significant DNA damage was observed by increasing in the index and frequency of damage along with the increasing of the frequency of micronuclei in peripheral blood and cells of the buccal mucosa, respectively. Despite the variability of the responses of breast cancer patients, the individuals presented lesions on the DNA, detected by the Comet assay and micronucleus Test, from the diagnosis until the end of the oncological treatment and were more susceptible to oxidative stress. We can conclude that the damages were due to clastogenic and/or aneugenic effects related to the neoplasia itself and that they increased, especially after RT.


Medicina ◽  
2007 ◽  
Vol 43 (1) ◽  
pp. 60 ◽  
Author(s):  
Vilma Jurkštienė ◽  
Anatolijus Kondrotas ◽  
Egidijus Kėvelaitis

The aim of the study was to investigate the immunostimulatory properties of bigroot geranium. Material and methods. Possible nonspecific characteristics of bigroot geranium were evaluated by the total leukocyte count in the peripheral blood, and qualitative changes of blood were assessed using Shilling’s formula by evaluating changes in lymphocyte counts. In addition, we also studied changes in the counts of Tcell precursors in the thymus and B lymphocytes in the spleen. Ethanol extract of the leaves of bigroot geranium was produced at the Department of Food Technology, Kaunas University of Technology. Studies were performed on mice Bl 57 (n=21). The control group (n=7) received distilled water at a dose of 1 mL/day. The second and third groups received 1% and 10% extract of bigroot geranium, respectively, as a food supplement. Changes in cell counts were investigated after 4 weeks following the initiation of the trial. Results. After a 4-week administration of 1% extract of bigroot geranium (1 mL/day) (mice group, n=7), leukocyte count in the peripheral blood increased to 6.1×109 cells/L, and lymphocyte count – to 70%, but changes were not statistically significant. The other case group of mice (n=7) received 10% extract of bigroot geranium for 4 weeks at a dose of 1 mL/day. In this group, leukocyte count in the peripheral blood increased statistically significantly from 4.4×109 cells/L to 7.2×109 cells/L (p<0.01), and lymphocyte percentage – from 52% to 80% (p<0.001), as compared to control. Thymocyte (T lymphocytes) counts in thymus and splenocyte (B lymphocytes) counts in the spleen showed a tendency to increase after the administration of 1% and 10% extracts. After a 4-week administration of 1% extract of bigroot geranium, thymocyte and splenocyte counts increased from 0.342×106 cells to 0.372×106 cells per mg of tissue and from 0.395×106 cells to 0.405×106 cells per mg of tissue, respectively, as compared to control group (p>0.1). After the administration of 10% extract of bigroot geranium, thymocyte count increased to 0.488×106 cells per mg of tissue (p<0.01), and splenocyte count – to 0.504×106 cells per mg of tissue (p<0.01). Conclusion. The extracts of the leaves of bigroot geranium increased leukocyte count and lymphocyte percentage in the peripheral blood, and after a 4-week administration of 10% extract of bigroot geranium, a statistically significant increase in the counts of T lymphocytes (in the thymus) and B lymphocytes (in the spleen) was observed. The immunostimulatory effect depends on the dose of the extract.


Animals ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 731
Author(s):  
Eduardo Naranjo-Vázquez ◽  
María Guadalupe Sánchez-Parada ◽  
Belinda Claudia Gómez-Meda ◽  
Ana Lourdes Zamora-Perez ◽  
Martha Patricia Gallegos-Arreola ◽  
...  

SKH1 hairless mice are widely used in carcinogenesis and dermatology research due to their bare skin, as exposure to different agents is facilitated. Minoxidil is a cosmetic drug that is recognized as a mitogenic agent, and mitogens are suggested to have carcinogenic and mutagenic potential by inducing cell division and increasing the possibility of perpetuating DNA damage. Therefore, we hypothesized that the application of high doses of minoxidil to the skin of hairless mice would increase the number of micronucleated erythrocytes (MNEs) in peripheral blood. The objective of this study was to evaluate the topical administration of high doses of minoxidil on peripheral blood erythrocytes of SKH1 mice by means of micronucleus assay. Minoxidil was administered on the entire body surface of mice every 12 or 24 h. Minoxidil dosing every 24 h increased the number of micronucleated polychromatic erythrocytes (MNPCEs), and dosing every 12 h increased the number of MNEs and MNPCEs, as compared to baseline and the negative control group. No decrease in polychromatic erythrocyte frequencies was observed in the minoxidil groups. Therefore, topical application of high minoxidil doses to mice can produce DNA damage, as observed through an increase in the number of MNEs, without producing cytotoxicity, possibly due to its mitogenic effect.


2013 ◽  
Vol 82 (3) ◽  
pp. 337-342
Author(s):  
Viera Revajová ◽  
Zuzana Slaminková ◽  
Ľubomíra Grešáková ◽  
Mikuláš Levkut

Morphometry and flow cytometry for intraepithelial lymphocyte phenotyping were used to determine the changes in duodenal mucosae after administration of low doses of deoxynivalenol in chickens. Moreover, functions of phagocytes and immunocompetent cells in peripheral blood were evaluated by flow cytometry. In total, sixty chickens of Ross hybrid broilers 308 were used in this experiment. Two experimental groups of 20 birds were continually fed for 14 days a diet containing deoxynivalenol at a dose of 1 and 3 mg·kg-1; 20 birds of the control group were fed uncontaminated diet. Morphometry showed only tendency to decrease the height of villi and surface area of duodenal mucosae in chickens fed the diet supplemented with 3 mg·kg-1 deoxynivalenol. Phenotyping of intraepithelial lymphocytes showed a decrease of CD45+ (P < 0.034) in duodenum of birds fed diets supplemented with deoxynivalenol. Examination of white blood cells showed a decrease of monocytes (P < 0.020) in chickens fed 3 mg·kg-1 of deoxynivalenol. Both experimental groups revealed higher metabolic burst of peripheral blood heterophils (P < 0.001). Phenotyping of immunocompetent cells showed an increase (P < 0.003) of CD3+ and a decrease (P < 0.001) of MHC II+ cells in peripheral blood of chickens fed with 3 mg·kg-1 dose of deoxynivalenol. The experimental feeding of chickens with deoxynivalenol resulted in immunomodulation of immunocompetent cells in duodenum and blood with mild atrophy of intestinal villi, mainly after the feeding of the dose of 3 mg·kg-1. We proved that even low doses of deoxynivalenol can cause changes in haemathological, immunological and morphological profiles already during two weeks, and lead to the activation of compensatory-adaptive mechanisms with unfavourable impact on health and performance of birds.


2021 ◽  
Vol 18 (4) ◽  
pp. 863-868
Author(s):  
Wanfeng Wu ◽  
Chengting Jiang ◽  
Cheng Cheng ◽  
Yihang Sun ◽  
Ning Luo ◽  
...  

Purpose: To study the combined effects of tenofovir and interferon α1b on viral load and peripheral blood regulatory T cell concentrations of chronic hepatitis B (CHB) subjects. Methods: Patients with chronic hepatitis B (86 cases) were randomly assigned to two groups: control group and study group. In control subjects, tenofovir was given orally (300 mg/kg bwt/day). In addition to tenofovir, the study group received interferon α1b injection intramuscularly at a dose of 50 μg/kg thrice a week. Liver function, serum hepatitis B viral (HBV) load, and serum levels of peripheral blood regulatory T-lymphocytes were determined. Clinical effectiveness and adverse reactions in both groups were also assessed. Results: After treatment, total effectiveness was higher in the study group (86.04 %) than in control patients (62.79 %) (p < 0.05). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and total bilirubin (TBIL) significantly decreased in the study group, relative to control, but HBV DNAnegative, HbeAg-negative and HbsAg-negative cells were markedly higher in patients in the study group (p < 0.05). Moreover, there were higher CD4+ T and CD8+ T counts, and CD4+ T/CD8+ T ratio in study subjects than in control subjects (p < 0.05). Conclusion: The combination of tenofovir with interferon α1b effectively improves liver functions in patients with CHB, reduces viral load, and exerts anti-HBV effect by regulating the levels of peripheral blood T-lymphocytes.


2021 ◽  
Vol 67 (6) ◽  
pp. 39-49
Author(s):  
M. A. Dudina ◽  
S. A. Dogadin ◽  
A. A. Savchenko ◽  
V. D. Belenyuk

BACKGROUND: Effective control of autoimmune inflammation in Graves’ disease determines necessity to study the T helper (Th) and cytotoxic T-lymphocytes dysfunction, as well as the level of regulatory T-cells (Treg) activation in patients with Graves’ disease on thyrostatic medication, which will clarify the immunomodulatory effects of long-term thiamazole treatment serve as targets for more specific therapies.AIM: To study the phenotypic composition of T-lymphocytes in the peripheral blood of patients with Graves’ disease to assess the direction of immune response depending on thimazole-induced euthyroidism duration.MATERIALS AND METHODS: A single-center, cohort, continuous, open-label, controlled trial was conducted to assess the phenotypic composition of T-lymphocytes in peripheral blood in women with Graves’ disease on long-term thiamazole treatment. The phenotypic composition of T-lymphocytes was determined by flow cytometry using direct immunofluorescence with conjugated FITC monoclonal antibodies depending on the duration of thimazole-induced euthyroidism of long-term thiamazole treatment.RESULTS: The study included 135 women with Graves’ disease, mean age 43.09±12.81 years, 120 (88.91%) with a relapse of the disease and 15 (11.09%) with newly diagnosed hyperthyroidism. An increase of activated CD3+CD4+CD25+ was found in patients with Graves’ disease with a duration of thimazole-induced euthyroidism 5–8 months and 9–12 months, respectively, Me=0.94 (0.48–1.45), p=0.020) and Me=0.95 (0.41–1.80), p=0.025), in control group — Me=0.12 (0.03–0.68). Compared to the control an increase of CD4+CD25+CD127Low (Treg) was found in patients with a duration of thimazole-induced euthyroidism 5–8 and 9–12 months. The content of Treg in peripheral blood in Graves’ disease patients with a duration of thimazole-induced euthyroidism more than 12 months decreases, but remains elevated relative to the control.CONCLUSION: In patients with Graves’ disease with a duration of thimazole-induced euthyroidism 5–8 months and 9–12 months the level of Treg has been increased. The increase of activated Th (CD3+CD4+CD25+) persists independently of thimazole-induced euthyroidism. In patients with Graves’ disease with a duration of thimazole-induced euthyroidism for more than 12 months, there is a compensatory increase in regulatory T-lymphocyte, and the total number of T-helpers is restored to the control.


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