Absolute Number Of Lymphocytes Research Articles

Immune system dysfunction in personnel of chemical hazard facilities: purpose and effectiveness of immunotherapeutic drugs

Clinical signs of the presence of immune dysfunction/insufficiency, identified by questioning workers at the landfill for the storage and destruction of industrial toxic waste, were the basis for performing a screening study of immune status. This included a clinical blood test with determination of the leukocyte formula, assessment of the phenotypic status of lymphocytes for the main subpopulations, determination of the concentration in serum immunoglobulins of various classes, assessment of the migration activity of leukocytes, assessment of the microbicidal capacity of neutrophils and their ability to phagocytosis. When the examined employees were found to have more than 20% changes in the normative range of immune status parameters and the presence of combined disorders in different parts of immunoreactivity, in particular combined disorders in the cellular part of the immune system, it was the basis for conducting an in-depth stage of immunoepidemiological examination. This included determination of functional activity cellular elements of the adaptive component of immunity, assessment of the severity of autosensitization, assessment of the severity and specificity of atopic hypersensitization, assessment of cytokine status in normal conditions and in response to mitogen. When assessing the phenotypic status of immunoreactivity cells, it turned out that the absolute number of T lymphocytes (CD3+ phenotype), helper T lymphocytes (CD3+CD4+ phenotype) and killer T lymphocytes (CD3+CD8+ phenotype) was significantly reduced in the peripheral blood of test site employees (p ≤ 0.05, compared with similar indicators in the control group). On the contrary, the content of NK cells (CD3-CD56+ phenotype) tended to increase (both the absolute number and the proportion among other lymphocyte populations). Dysfunction of the cellular component of humoral immunoreactivity (a significant decrease in the absolute number of lymphocytes with the CD3-CD20+ phenotype [B lymphocytes] in the peripheral blood), as well as a significant increase in the concentration of serum IgA-class immunoglobulin and pronounced disimmunoglobulinemia in the serum of the examined individuals (compared to individuals in the control group ) indicated a predominantly inhalation method of exposure to immunotoxic industrial toxicants from the landfill on the immune system. Pathogenetically targeted immune-oriented therapy for the personnel of the test site was carried out with drugs that have thymomimetic activity (cytovir-3, thymogen) and are a means of replacement immunocorrection with a cytokine drug (genetically engineered recombinant yeast IL-2 [roncoleukin]). The established effectiveness of the immunotherapy, in particular the reduction in the incidence rate of various nosological forms of immune-related pathology among the site employees who received immunotherapy and the presence of an immunocorrective effect in the immunoactive drugs used for therapy, indicate the advisability of carrying out preventive medical measures in persons professionally exposed to xenobiotic influences as a component of improving medical provision.

Expression of γδ T cells and lymphocyte subsets in newly diagnosed multiple myeloma patients

ObjectiveTo retrospectively analyze the expression and significance of γδ T cells and lymphocyte subsets in peripheral blood on patients with multiple myeloma (MM). MethodsFifty-five patients with multiple myeloma admitted to Yangpu Hospital Affiliated to Tongji University from January 2022 to August 2023 were selected as the disease group, and were categorized into stage I group, stage II group and stage III group according to the International Staging System (ISS) of multiple myeloma. All included patients were consecutive visits. Another 30 cases of healthy people with physical examination were selected as the control group. Flow cytometry was used to detect the γδ T cells and lymphocyte subsets in the peripheral blood of the subjects. After clinical indicators of patients and healthy controls were collected, the statistical analysis was conducted. ResultsCompared with the control group, the percentage of γδ T cells and CD8+ T cells in the disease group were higher, while, the absolute numbers of total peripheral blood lymphocytes (LYMT) and CD19+ B cells, percentage of CD19+ B cells, CD3+ T cells, CD4+ T cells were lower (P​<​0.05). The percentage of γδ T cells (P​<​0.001), the percentage of CD19+ B cells (P​=​0.003), the absolute number of CD3+ T cells (P​=​0.010), the absolute number of CD8+ T cells (P​=​0.002) were the significant influential factors of MM. γδ T cells (P​=​0.004), neutrophils (P​<​0.001), neutrophil to lymphocyte ratio (P​=​0.003), and white blood cell count (P​<​0.001) were statistically different in different stages. ROC curve suggested that the area under the curve of the percentage of γδ T cells in predicting the MM were 0.973 (P​<​0.001) which had certain diagnostic value. Conclusionsγδ T cells and lymphocyte subsets (CD8+ T cells, CD19+ B cells, CD3+ T cells, etc.) in patients with newly diagnosis of MM can reflect the immune function of the patient’s organism, which is a key factor of MM. γδ T cells influence the staging of MM patients, and also have certain diagnostic value.

Peculiarities of the blood system of animals under the conditions of consumption of drinking water containing chlorates

In modern conditions, providing the population with high-quality drinking water is becoming an increasingly urgent hygienic, scientific, technical, and social problem. Chlorine dioxide can be an alternative reagent for disinfecting drinking water. However, as with chlorine, dangerous by-products, particularly chlorites and chlorates are formed. The processes of development of morphofunctional changes in homeostasis remain insufficiently studied in this aspect. The objective is to determine the nature and peculiarities of changes in hematological indicators in the blood of animals due to the chronic impact of various concentrations of chlorates in drinking water to establish criteria for the hygienic assessment of their adverse effects on the body. The test animals were given drinking water with different concentrations of chlorates daily for 6 months: group 1 – control, group 2 – chlorates at a dose of 0.2 mg/dm³, group 3 – chlorates at a dose of 0.7 mg/dm³, and group 4 – chlorates at a dose of 1.2 mg/dm³. Evaluating the results of experimental studies, it should be noted that the most pronounced changes are observed in groups of animals that received chlorates in concentrations of 0.7 and 1.2 mg/dm³: a gradual decrease in the absolute number of leukocytes, the absolute and relative number of lymphocytes, relative to the indicators of the control group can be observed. In groups of animals that received chlorates in concentrations of 0.7 and 1.2 mg/dm³, a decrease in the absolute number of erythrocytes was observed during the 90 days of the experiment. It was established that the severity of the effects depended on the dose of chlorates and the time of exposure. Thus, the nature and features of changes in hematological indicators in the body of experimental animals during a chronic sanitary-toxicological experiment under the influence of chlorates in drinking water may indicate that chlorates affect the indicators of the blood system, being the indirect signs of disorders of other functional systems in the body, namely – slowing of redox reactions, hypoxic manifestations, reduction and weakening of the immune response and reactivity, etc.

Blood smears analysis: a valuable tool for profiling circulating erythrocytes and leukocytes in mouse model of leishmaniasis

Although blood smear analysis can yield valuable insights into the health status of laboratory animals, it is not frequently performed in studies involving mouse models of leishmaniasis. Thus, to underscore its importance, we conducted microscopic evaluation of blood smears prepared from 10 µL of blood from L. (L.) amazonensis-infected BALB/c mice. Subsequently, we explored potential correlations between the identified hematological alterations, clinical evaluation data, and serum levels of total proteins (TP), albumin (Alb), globulins (Glob), IL-17A, and IL-10. Our findings revealed that infected mice exhibited an increased erythrocyte count, alongside a higher prevalence of microcytic and hypochromic erythrocytes. Moreover, it was observed elevated relative and absolute numbers of neutrophils and monocytes and reduced relative and absolute numbers of lymphocytes. A significant proportion of monocytes and lymphocytes in infected animals displayed reactive features. The relative number of lymphocytes was negatively correlated with the serum levels of Glob. Furthermore, the frequency of reactive lymphocytes was positively correlated with the serum levels of Glob and IL-17A. There was also a positive correlation between the absolute number of neutrophils and the serum levels of IL-17A and the frequency of reactive monocytes and the serum levels of Glob. Therefore, these findings, collectively, underscore the importance of blood smear analysis for elucidating both the quantitative and qualitative hematological changes experimentally induced by Leishmania infection. Integration of this technique into research protocols involving different mouse models of leishmaniasis holds immense potential for advancing our knowledge of this infectious disease and enhancing clinical management strategies.

#1815 The neutrophil/lymphocyte, neutrophil/HDL-cholesterol, and monocyte/HDL-cholesterol ratios as simple markers of low-grade inflammation in CKD patients

Abstract Background and Aims Chronic kidney disease (CKD) has been associated with inflammation and oxidative stress, as well as with high cardiovascular (CV) morbidity and mortality. The neutrophil/lymphocyte (NLR), neutrophil/HDL-cholesterol (NHR), and monocyte/HDL-cholesterol (MHR) ratios have been linked both to inflammation and increased CV risk in a variety of diseases. The aim of this study was to investigate changes in these ratios in CKD patients of stage 2-5 without any clinical and laboratory signs of inflammation, and to compare these indicators with those in healthy volunteers. Method A retrospective cross-sectional study was conducted in 88 healthy volunteers and 187 CKD patients (50 of stage 2, 44 of stage 3a, 45 of stage 3b and 48 of stage 4-5). Patients with acute renal impairment, nephrotic proteinuria, infections, systemic inflammation, malignancies and hematologic diseases were excluded, as were those receiving hypolipidemic, corticosteroid, immunosuppressive or immunostimulant drugs. The whole blood samples were collected and the absolute numbers of neutrophils, lymphocytes, and monocytes were used for calculation of NLR, NHR and MHR, respectively. Serum creatinine, HDL-cholesterol, C-reactive protein (CRP) and albumin were measured and eGFR calculated according to the CKD EPI equation. Statistical analysis was performed with the Mann-Whitney, Shapiro–Wilk, Kruskal-Wallis, and Spearman's correlation tests using the SPSS software. Results The NLR, NHR and MHR were higher in CKD patients when compared to healthy volunteers (P &amp;lt; 0,001), but only NLR and NHR increased significantly across CKD stages (P &amp;lt; 0,01). The ratio differences between CKD stages were more evident for NHR which was also in best correlation with eGFR (ρ = −0,406, P &amp;lt; 0,001). The CRP values increased across CKD stages as well (P &amp;lt; 0,01) although remaining within the reference range. The NLR, NHR and MHR were significantly related to CRP with NHR providing the best correlation (ρ = 0,289, P &amp;lt; 0,001). Conclusion This study showed that NLR and NHR increased across CKD stages 2-5 and could be used to identify low-grade inflammation in this population even within normal CRP values.

Value of Platelet to Lymphocyte Ratio in Nasopharyngeal Carcinoma Various Stages.

Nasopharyngeal Carcinoma (NPC) is among the most common head and neck cancer, significantly affecting the growth of human tumors. Therefore, this study aimed to investigate platelet-to-lymphocyte ratio (PLR) as a measure of inflammation in patients with various stages of NPC. This retrospective case-control study was conducted on patients diagnosed with NPC between 2016 and 2020 in Dr. Hasan Sadikin Bandung Hospital, Indonesia. Based on NPC classification criteria, patients were classified into early and advanced groups. Subsequently, the PLR ratio was calculated as the number of platelet divided by the absolute number of lymphocytes in the two groups, and the results were statistically analyzed using Unpaired T-test, Pearson, and Chi-square test. A total of 102 patients with an age range of 40 to 49 years were included in this study. A slight correlation of 0.272 (P<0.001) was observed between PLR and clinical stage, while NPC patients with PLR value > 188 had a 2.7 times higher risk of advanced stage (RR=2.702). The PLR was significantly related to the clinical stage of NPC cancer.

Morphological Characteristics of Colon Tumors in Mice with Different Tolerance to Hypoxia.

In adult male C57BL/6 mice with high (HR) and low (LR) resistance to hypoxia, morphological features of colon tumors and blood parameters were evaluated 70 days after intraperitoneal injection of azoxymethane and subsequent consumption of 3 cycles of dextran sulfate sodium. On macroscopic analysis, tumors were found in the distal colon in 35% (7 of 20 animals) of HR and 31% (4 of 13 animals) of LR animals. Microscopic analysis of the distal colon revealed tumors in 75% (15 of 20 animals) of HR and 69% (9 of 13 animals) of LR mice. The tumors were presented by areas of glandular intraepithelial neoplasia and adenocarcinomas; the incidence and the area of the tumors did not differ in groups of HR and LR mice. The number of neuroendocrine and goblet cells in the distal colon mucosa in the areas of tumors was similar in the compared groups. However, in both HR and LR mice of the experimental groups, the content of goblet cells in tumors was lower and the content of endocrine cells was higher than in the corresponding control groups. In the peripheral blood, the erythrocyte count and hemoglobin content decreased in HR and LR mice of the experimental groups; the relative number of monocytes increased only in HR mice and the absolute number of lymphocytes and monocytes decreased in LR mice. Thus, 70 days after azoxymethane administration and dextran sulfate sodium consumption, the tumors in mice were presented by glandular intraepithelial neoplasia and adenocarcinomas, and their incidence and area did not differ between animals with different tolerance to hypoxia.

Early “Red Flags” in hospitalized patients with COVID-19

Relevance. Among the studies during the COVID-19 pandemic, a special role is played by studies examining the prognostic value of clinical, laboratory and instrumental markers in patients. Biomarkers with absolute sensitivity and specificity for predicting the outcome of a new coronavirus infection have not been identified yet.Aim: To identify early clinical and laboratory predictors of severity and mortality in hospitalized patients with COVID-19 and to evaluate the effectiveness of the pharmacotherapy in outpatient and early hospital settings.Methodology and research methods. A retrospective, comparative study was carried out. Medical data from 745 adult patients with COVID-19, hospitalized in the respiratory hospital of the Siberian State Medical University (Tomsk) from 05/16/2020 to 09/30/2020 were included. The main group consists of patients who were discharged from the hospital. The comparison group included patients with a fatal outcome of the disease.Results. The group of deceased patients was characterized by older age and a greater number of comorbid diseases (р &lt; 0.0001), the presence of pneumonia during the year preceding COVID-19 (р &lt; 0.0001) and risk factors for the development of Pseudomonas aeruginosa infection (р &lt; 0.005). Early admission to the hospital (р = 0.0110), severe dyspnea (р &lt; 0.0001) with a decrease in oxygen saturation below reference values (р &lt; 0.0001) are also associated with the risk of a fatal outcome. Anosmia was significantly more common in the group of surviving patients.The number of platelets, the absolute number of lymphocytes, amounts of total protein, albumin (р &lt; 0.0001), sodium (р = 0.0013) in the first 48 hours of hospitalization in the main group were significantly higher. The number of leukocytes (p = 0.0078), neutrophils (p &lt; 0.0001), neutrophil-lymphocyte ratio (p &lt; 0.0001), concentrations of glucose, aspartate aminotransferase, urea, creatinine, C-reactive protein and lactate dehydrogenase were significantly lower in the main group. Serum procalcitonin levels &gt; 0.5 ng/ml were 5 times higher in deceased patients during early period of hospitalization.Conclusion. Clinical, anamnestic and laboratory factors were identified through this study. These factors may be useful in the early period of hospitalization for accurate assessment of risks of severe ill, effective disease management and reduction of mortality in hospitalized patients with COVID-19.

The effects of aging, sex, and tumor burden on the peripheral blood immune cell profile and absolute counts.

Background: To better assess the peripheral immune status and aid in the early diagnosis and prognosis of tumors, we compared the proportion and absolute counting of peripheral immune cell subsets in healthy individuals and tumor patients of varying ages, taking into account the impact of sex and tumor metastasis. Methods: We used peripheral blood mononuclear cell (PBMC) samples from 520 patients with various tumor types and 109 healthy volunteers. The absolute numbers of lymphocytes and monocytes were identified by an automated blood analyzer, and multi-parameter flow cytometry was used to examine the subsets of natural killer (NK) cells (CD3-CD16+CD56+), T cells (CD3+CD4+/CD8+), and mononuclear cells (CD14+) in PBMC. Results: The percentage of T cells (CD3+) in peripheral blood mononuclear cells (PBMC) was 55.83% VS 45.54% (P<0.0001) between healthy volunteers and tumor patients, showing a significant downward trend. Meanwhile, the percentages of monocytes (CD14+) and NK cells (CD3-CD16+CD56+) showed a significant upward trend. Single factor or multifactor analysis yielded identical findings on the proportion of PBMC between healthy individuals and patients with different malignancies, considering the three confounding variables of age, sex, and tumor metastasis. Conclusion: The proportion and absolute counting of acquired immune T cells, innate immune NK cells, and monocytes in PBMCs all exhibit substantial changes between cancer patients and healthy individuals, and the differences are influenced by age, sex, and tumor progression.

Multi-strain probiotic combined with dietary fiber is an effective factor in the nutritional support of immunity in athletes

A priority in the prevention and correction of immune disorders in athletes is the use of products with high nutrient density, fortified with various nutrients or bioactive compounds, as well as probiotic microorganisms. Probiotics help to maintain the gut microbiota, which is actively involved in the absorption of substances and energy and increases the host immune resistance. Dietary fiber, resistant to digestion in the small intestine, is fully or partially fermented in the large intestine and acts as an essential substrate for the growth and regulation of metabolic activity of normal flora, improves peristalsis and digestion. The purpose of the study was to evaluate the impact of a multi-strain probiotic in combination with dietary fiber on the immune status of basketball athletes during the training period. Material and methods. The study was conducted with the participation of 30 male basketball athletes aged 18 to 24 years. The athletes were randomly divided into 2 groups of 15 people. Athletes in the main group received 1 capsule of multi-strain probiotic (≥1.25×1010 CFU of 10 probiotic strains of bifidobacteria and lactobacilli) and 40 g of corn bran (as a source of dietary fiber) for 23 days. Athletes in the control group received 1 placebo capsule containing maltodextrin and breadcrumbs (40 g/day). Subpopulations of peripheral blood lymphocytes were studied by flow cytometry: T lymphocytes, T helper cells, T cytotoxic lymphocytes, NK cells, NKT cells, B lymphocytes, as well as lymphocytes carrying activation markers and apoptosis marker antigen. The content of cytokines in blood serum [FGF, Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1ra, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12(p70), IL-13, IL-15, IL-17A, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α, VEGF] was determined using a multiplex immunoassay. Results. Calculation of the absolute number of lymphocytes revealed a tendency (0.05<p<0.10) to a decrease in T helper cells by the end of the observation period in athletes of the main group (497.60±27.67 vs 632.67±65.20 cells/μL), as well as a decrease (p<0.05) in the expression of the CD95/Fas apoptotic marker on peripheral blood lymphocytes of athletes of the main group compared to the beginning of the study (41.53±5.78 vs 69.53±11.79 cells/μL). At the end of the study, a significant increase in IL-9 level was found in the control group [(Me; min-max) = (0.33; 0.21-0.48) vs (0.26; 0.09-0.38) pg/ml; p<0.05; in comparison with the initial indicator]; as well as the tendency (0.05<p<0.10) towards an increase in the levels of IL-15, IL-1ra and RANTES was revealed. In the main group, at the end of the study, the level of G-CSF significantly decreased [(0.36; 0.03-0.95) vs (0.53; 0.14-1.36) pg/ml, p<0.05]. At the end of the observation period, blood serum levels of FGF, G-CSF, IL-13, IL-2 and RANTES in the athletes of the control group exceeded these indicators in the athletes of the main group whereas no significant differences in the studied cytokines were detected between the control and the main groups at the beginning of the study. Conclusion. The results of observation of basketball athletes who consumed a multi-strain probiotic in combination with corn bran (sources of arabinoxylans) in addition to the main diet for 23 days indicate a decrease in the inflammatory process activity and peripheral blood lymphocyte apoptosis, which confirms the effectiveness of probiotics and dietary fiber in sports nutrition.

PATHOGENESIS OF IMMUNE DYSFUNCTION IN THE SETTING OF METABOLIK SYNDROME IN CORONARY HEAT DISEASE. QUALIFYING SCENTIFYING WORK ON THE RIGHTS OF THE MANUSCRIPT

PATHOGENESIS OF IMMUNE DYSFUNCTION IN THE SETTING OF METABOLIK SYNDROME IN CORONARY HEAT DISEASE. QUALIFYING SCENTIFYING WORK ON THE RIGHTS OF THE MANUSCRIPT

The role of clinical and immunological parameters in predicting the effectiveness of additional immunotropic therapy in oxygen-dependent patients with COVID-19 coronavirus disease

Aim. The purpose of the research is to find out the role of clinical and immunological parameters in predicting the effectiveness of additional immunotropic therapy in oxygen-dependent patients with the coronavirus disease COVID-19. Material and methods. 79 oxygen-dependent patients with COVID-19 were examined, who received additional therapy with tocilizumab according to current protocols. The patients were divided into groups: I group – 39 patients who recovered; group II – 40 patients with a fatal outcome of the disease. The content of ferritin (Monobind Inc., USA), interleukin (IL) IL-2 (Elabscience, USA), IL-6 (Invitrogen, Austria), monocyte chemotactic protein-1 (MCP-1) (Elabscience, USA) was determined in the blood serum of patients and persons of the control group by the method of immunoenzymatic analysis. Statistical processing of the obtained data was carried out in the program Statistica 13 for Windows (StatSoft Inc., No. JPZ804I382130ARCN10-J). Results. Threshold levels of immunological parameters at different stages of observation, which have prognostic value regarding the risk of a fatal outcome of COVID-19, have been established. After 5 days of tocilizumab administration, the preservation of the indicator of the absolute number of blood lymphocytes ≤1.2 × 109/l (AUC = 0.631, p = 0.039), CRP level &gt;67.5 mg/l (AUC = 0.670, p = 0.020), IL-2 &gt;309.45 pg/ml (AUC = 0.761, p = 0.013), IL-6 &gt;4.66 pg/ml (AUC = 0.871, p &lt; 0.001) indicated a high probability of developing a fatal outcome of the disease in patients with COVID-19. The analysis of the diagnostic significance of ferritin showed the informativeness of the level of its increase as at the time of the start of observation &gt;548.02 ng/ml (AUC = 0.718, p = 0.004) and after 5 days the application of additional immunotropic therapy &gt;443.55 ng/ml (AUC = 0.736, p = 0.026). It has been proven that, in addition to immunological parameters, when assessing the probability of a fatal outcome of COVID-19, such clinical parameters as the presence of febrile fever at the time of the appearance of oxygen dependence (p &lt; 0.05) and the duration of oxygen dependence at the time of additional immunotropic therapy with tocilizumab are informative (p &lt; 0.05). Under the conditions of additional immunotropic therapy with tocilizumab more than 4 days after the onset of oxygen dependence, the probability of ineffectiveness of the specified additional treatment was significant (AUC = 0.756, p &lt; 0.001). Conclusions. The dynamics of immunological parameters in oxygen-dependent patients with COVID-19 during additional immunotropic therapy with tocilizumab has certain features with different treatment results. When predicting the probable risk of a fatal outcome of the disease, clinical parameters are informative, namely the appearance of febrile fever during the development of oxygen dependence and the duration of oxygen dependence until the moment of tocilizumab administration, and immunological parameters, namely the level of the absolute content of lymphocytes, ferritin, CRP, IL-2, IL-6 in the blood.

The state of cellular immunity link and steroid hormones in bitches under different conditions of the reproductive system

The purpose of the conducted study was to establish the correlation between indicators of the cellular immunity link and the concentration of steroid hormones in bitches under different conditions of the reproductive system. Seventeen crossbred bitches aged from 2 to 5 years were involved in the experiment. They were divided into three groups depending on the state of the reproductive system: anestrus, sexual cycle, and pyometra. The following immunophysiological indicators were determined in blood plasma: the absolute number of leukocytes, lymphocytes, and their immunoregulatory subpopulations, and the phagocytic activity of neutrophils. The content of progesterone, estradiol, and cortisol was determined in serum. According to the results of the research, it was established that the absolute number of neutrophils, as the most reactogenic cells of the innate link of cellular immunity, was 0.51 G/L higher in the group of animals with pyometra than in the group of animals during the sexual cycle, and by 0.84 G/L in bitches in conditions of anestrus. The absolute number of phagocytic active neutrophils in conditions of pyometra was 2.32 ± 0.58 G/L (Р &lt; 0.001). However, as a percentage of the absolute number of neutrophils, this indicator was 66 %. During the sexual cycle, the absolute number of neutrophils capable of phagocytosis was 2.33 ± 0.51 G/L, which is 78 % of the absolute number of all neutrophilic leukocytes. The indicator of the absolute number of lymphocytes was reliably (P &lt; 0.05) lower in the group of bitches with pyometra – 1.48 ± 0.35 G/L, which is 0.92 G/L less compared to the group of animals in the anestrous period. Analysis of the absolute number of monocytes showed that this indicator was the same in bitches during the pyometra and anestrous periods and was lower by 0,07 G/L during the sexual cycle. The absolute number of B-lymphocytes, as a humoral link of adaptive immunity, in bitches in conditions of the anestrous and estrous period was the same and amounted to 0.30 G/L. In the group of bitches with pyometra, a reliable decrease of this indicator by 33 % was established (Р &lt; 0.05). The population of natural killer (NK) lymphocytes was also the lowest in bitches with pyometra, almost twice as compared to animals during the sexual cycle. Conducted research on determining the absolute quantitative indicators of the cellular immunity link showed their dependence on the functional state of the reproductive system. It was established that the absolute number of neutrophils in pyometra was higher than in animals in anestrus. However, at the same time, their relative number capable of phagocytosis was the lowest. In conditions of pyometra, the lowest number of lymphocytes and their main subpopulations were observed.

The particularities of SARS-CoV-2 infection on onco-hematological diseases in children

The particularities of SARS-CoV-2 infection on onco-hematological diseases in children. Introduction: Cancer in childrenis a rare disease among the pediatric population, representing approximately 0.5%- 1% of the total number of cancers. In the Republic of Moldova, anually, approximately 90 children are diagnosed with malignant tumors. The survival rate of children with cancer has improved considerably over time, but it remains a significant cause of morbidity and mortality among children. In oncological pathology, viral and bacterial infections acquired in the community can lead to a delay in chemotherapy treatment, which increases the risk of disease recurrence and resistance, thus increasing mortality. Children with hematologic malignancies generally have a more favorable prognosis for COVID-19 infection than adults with the same pathologies, but their risks of hospitalization, and death are much higher compared to the general pediatric populationThe purpose of the study: Determination of the infection rate of children with hemoblastosis with SARS Cov 2 infection, evolution and survival rate. Materials and methods: Thisis a retrospective study carried out in the Oncological Institute, hematology section for children, carried out during the SARS-CoV-2 pandemic March 2020-March 2022. Results: Among all patients hospitalized and treated in theOncological Institute, the children’s hematology section (125 patients) - 33 (26.4%) patients suffered SARS COV 2 infection, of which 3 had endured covid 2 times. The majority of patients - 26 patients (78.78%) had acute lymphoblastic leukemia, 1 (3.03%) Hodgkin’s lymphoma and 2 (6.06%) with Burkitt’s lymphomas, and non-Hodgkin’s lymphoma. Among the clinical symptoms, most children presented with 11 patients (30.5%), and fever&gt; 38-10 patients (27.7%), 12 children (33.3%) did not present any symptoms, only the positive COVID-19 test. Among children diagnosed with COVID-19 infection, 10 (27%) cases of COVID were registered in children before or simultaneously with the basic hematological diagnosis, 25% (9 patients) were in the period of intermediate maintenance and we had one case ( 2.7%) in reinduction therapy and refractory therapy respectively. Analyzing the paraclinical data, a slight prevalence of the absolute number of lymphocytes compared to that of neutrophils was observed in patients who presented radiological signs of viral pneumonia. According to the imaging data, we had 7 confirmed cases of bilateral viral pneumonia, Rx1, 1 case of severe bilateral viral pneumonia-Rx4, which resulted in death, and one case of bilateral viral pneumonia due to leukemoid infiltration which also led to death, the child being decompensated approximately 2 months before the diagnosis of COVID-19. Conclusions: For most pediatric patients with hematologic malignancies, infection with COVID-19 does not result in severe progression. But this infection leads to the discontinuation of specific therapy, but the long-term consequences are notyet sufficiently studied.

Applying decision tree algorithms to early differential diagnosis between different clinical forms of acute Lyme borreliosis and tick-borne encephalitis

Background: Tick-borne encephalitis and Lyme borreliosis are the most common natural focal infections in Russia often arising as a mixed infection, which is often clinically difficult to distinguish from a monoinfection at the onset of the disease that is a result of delayed laboratory verification of the diagnosis and it requires further searching for fundamentally new approaches to the issue of early differential diagnosis of tick-borne infections.Aims: is to develop decision tree algorithms for early differential diagnosis between the mono- and mixed forms of acute Lyme borreliosis and tick-borne encephalitis with prevailing febrile syndrome in clinical picture based on clinical and laboratory data.Materials and methods: We retrospectively analyzed 55 clinical and laboratory parameters obtained from 291 hospitalized tick-borne infections patients with or without erythema migrans at the site of ixodid tick bites in the first week of the disease, who were included in the single-center study from 2010 to 2023. In 211 patients without erythema, the analysis was carried out between three classes depending on the diagnosis: the mixed infection of non-erythematous Lyme borreliosis and tick-borne encephalitis, the mono-infection of non-erythematous Lyme borreliosis or the monoinfection of tick-borne encephalitis. The other two classes, which included 80 patients with erythema, had the mixed infection of acute erythematous Lyme borreliosis and tick-borne encephalitis or erythematous Lyme borreliosis monoinfection. Python programming language was applied to develop two decision tree models. Feature importance was assessed for all predictors. Each patient class was randomly divided into training (70%) and testing (30%) datasets. Accuracy evaluation of the models was based on ROC analysis.Results: The decision tree algorithm for early differential diagnosis among the tick-borne infection patients without erythema migrans included the following most important predictors: maximal fever rise, chills, neutrophil-to-monocyte ratio, ESR, absolute number of reactive lymphocytes and immature granulocytes, and percentage of eosinophils. The model for differential diagnosis between the patients with erythema migrans included the following predictors: maximal fever rise, the absolute number of reactive lymphocytes and immature granulocytes, and the percentage of basophils. Both decision tree models showed excellent predictive values based on sensitivity, specificity, precision, accuracy, and F1 scores, as well as areas under the ROC curve, which were higher than 0.90.Conclusions: Based on clinical and laboratory parameters, two decision tree models with high sensitivity have been developed, which can be easily applied in clinical practice for early differential diagnosis of the tick-borne infections with prevailing fever syndrome.

Systemic Inflammation Response Index Predicts Clinical Outcomes in Patients With Acute Ischemic Stroke (AIS) After the Treatment of Intravenous Thrombolysis.

Intravenous thrombolysis (IVT) is one of the most important means of therapy for patients with acute ischemic stroke (AIS). After cerebral infarction, the inflammatory response fulfills an essential role in the pathobiology of stroke, affecting the process of recanalization. Hence, we evaluated the usefulness of the systemic inflammatory response index (SIRI) for the prognosis of patients with AIS. A total of 161 patients suffering from AIS were retrospectively analyzed. SIRI was introduced and calculated using the absolute neutrophil, monocyte, and lymphocyte numbers from the admission blood work. The study outcomes were determined using a modified Rankin Scale (mRS) at the 3-month timepoint, and a favorable clinical outcome was calculated in the mRS score range of 0 to 2. The analysis of receiver operating characteristic (ROC) curves was performed to determine the values of the optimal cutoff of SIRI for the prediction of clinical outcomes. In addition, multivariate analyses were performed to investigate the association between clinical outcomes and SIRI. The ROC curve analysis revealed that the ideal SIRI cutoff was at 2.54 [area under the curve, 78.85%; 95% CI, 71.70% to 86.00%; sensitivity, 70.89%; and specificity, 84.14%]. Multivariate analysis indicated that SIRI ≤2.54 (odds ratio, 1.557, 95% CI, 1.269 to 1.840; P=0.021) was an independent predictor of favorable clinical outcomes in patients suffering from AIS after treatment with IVT. We preliminary speculate that SIRI may serve as an independent predictor of clinical outcomes with AIS following IVT.

HPV and Systemic Response in Cervical Intraepithelial Neoplasia

Objectives: to verify whether there is an association of low and high-risk HPV with blood count parameters in patients with cervical intraepithelial neoplasia. Methods: We prospectively evaluated 48 patients with Pap smear showing low or high-grade intraepithelial lesion, submitted to collection of material to perform PCR for high-risk and low-risk HPV. The following blood count parameters were evaluated: hemoglobin, red blood cells, hematocrit, absolute number of leukocytes, neutrophils, lymphocytes, eosinophils, basophils, monocytes and platelets, neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR) and Red Cell Distribution Width (RDW). Receiver operating characteristic (ROC) curve was used to obtain the area under the curve (AUC) relating the positivity or not of HPV with the blood count parameters evaluated. Mann-Whitney test and multivariate analysis were performed, with level of significance less than 0.05. Results: Relating the positivity or not of HPV with the blood count parameters (Roc curves), there was statistical significance only between high-risk HPV and the platelet count (sensitivity=75%, specificity=66.7%, AUC=0.674, p=0.039). Platelet count ≤ 177,000/mm3 is associated with high-risk HPV. Subsequently, the multivariate analysis was performed, and the absolute value of platelets was considered an independent variable in this association with high-risk oncogenic HPV. Conclusion: A systemic response is found with the presence of high-risk HPV in CIN. Thus, platelet count ≤ 177,000/mm3 is associated with high-risk HPV.

Evaluation of the toxicity of the hydroethanolic extract of the stem bark of Virola elongata (Benth.) Warb. in in vitro and in vivo models

Ethnopharmacological relevanceVirola elongata (Benth.) Warb. (Myrsticaceae), referred to as “mucuíba" in Mato Grosso, is a native tree species that can be found in the Amazon Rainforest regions of South America and the Tropical Forests of Central America. The macerated extracts from the stem bark of this tree have been traditionally used to address various health issues, including gastric ulcers, infections, inflammations, and other ailments. In scientific literature, V. elongata has demonstrated pharmacological properties such as antiulcer, gastroprotective, antiproliferative, antimitotic, and psychoactive effects. Nevertheless, it is important to note that the safety profile of V. elongata has not been thoroughly established. Aim of the studyTo evaluate the toxicity of the hydroethanolic extract of the stem bark of Virola elongata (HEVe) in experimental models in vivo and in vitro. Materials and methodsHEVe was obtained by macerating the stem bark powder in 70% hydroethanolic solution (1:10 w/v). The cytotoxicity of HEVe (3.125–200 μg/mL) was evaluated by Alamar blue assay in Chinese hamster ovary epithelial cells (CHO-k1) and human gastric adenocarcinoma (AGS). Genotoxicity assessment of HEVe (10, 30, or 100 μg/mL) was performed in CHO-k1 cells by the micronucleus test. The acute toxicity of HEVe was assessed by single-dose oral administration (2000 mg/kg) in mice of both sexes. The subacute toxicity of HEVe was assessed by oral administration of 300, 600, and 1200 mg/kg of the extract over 30 days in rats. Clinical observations of toxicological parameters were noted and pooled every 6 days. After the treatment period, blood was collected for hematological and biochemical analyses, and some organs were removed for macroscopic and histopathological analyses. ResultsHEVe did not show cytotoxicity in CHO-K1 and AGS cells (IC50 > 200 μg/mL) and did not cause DNA damage in CHO-k1 cells. Oral administration of HEVe in a single dose of 2000 mg/kg did not result in the death of the mice, with a reduction in body weight variation (33.03%, p < 0.05) and an increase in the relative weight of the stomach (12 0.82%, p < 0.05) in male mice, and increased relative weight of the spleen (25.00%, p < 0.01) in female mice. In the assessment of subacute toxicity, HEVe did not result in the death of the animals over the 30 days. A reduction (p < 0.05) in water consumption of 36.65% and 34.12% was observed in the groups treated with 300 and 600 mg/kg, respectively, of HEVe on D6., and the urine excretion of animals treated with 600 mg/kg of HEVe showed an increase (p < 0.05) throughout the experiment, with a maximum value of 46.72% on D12. The blood counts showed that the dose of 300 mg/kg reduced (p < 0.05) the absolute number of lymphocytes, while the doses of 300, 600, or 1200 mg/kg of HEVe reduced the red blood cell count in whole blood by 24.84% (p < 0.01), 16.72% (p < 0.05), and 22.14% (p < 0.01), and the absolute number of monocytes (p < 0.05) in 59.77%, 65.51%, and 79.81%, respectively. As for the biochemical parameters, the glucose level found increased by 22.41% (p < 0.05) only at the highest dose, while creatinine was reduced by 44.71% (p < 0.05) at the dose of 300 mg/kg of HEVe. In animals treated with the three doses tested, plasma levels of AST and alkaline phosphatase showed a reduction (p < 0.05) with the vehicle group. However, the hematological and biochemical changes observed are within the physiological limits for this animal species. No macroscopic and histopathological changes were observed in the organs of the animals treated with the three doses of HEVe within 30 days. ConclusionThe results showed that HEVe did not show cytotoxicity or genotoxicity in vitro. HEVe proved to be safe in rodents in both acute and subacute toxicity tests. In rats, the no-observed-adverse-effect level (NOAEL) dose was greater than 1200 mg/kg p. o. in rats.

Experimental study of quantitative and corpuscular properties of rat’s leukocytes under the influence of ore

Introduction. Mining industry workers are at risk, as heavy metal compounds that are part of copper-zinc pyrite ore can accumulate in their bodies. One of the sensitive parts of the body to the action of heavy metals is blood, in which leukocytes play a leading role.&#x0D; The study aims to explore the effect of copper-zinc pyrite ore on the quantitative and corpuscular properties of peripheral blood leukocytes.&#x0D; Materials and methods. A group of researchers have conducted an experimental study on 48 white mongrel male rats aged 3–4 months, weighing 210.5±10.5 g. For 30 days, an aqueous suspension of ore powder was orally administered to rats daily at a dose of 600 mg/kg of animal body weight. In the ore sample under study, the proportion of lead and cadmium was 0.060% and 0.009%, respectively. In accordance with the conditions of the experiment , the scientists have divided the rats into the following groups: a control group in which the rats did not receive ore; an experimental group in which rats received ore for 10 days; an experimental group in which rats received ore for 20 days; an experimental group in which rats received ore for 30 days. The researchers measured blood parameters using a veterinary semi-automatic hematology analyzer Vet Exigo 19 (Sweden), which determined the following parameters of leukocytes: total number of leukocytes, absolute number of lymphocytes, monocytes and granulocytes. We calculated the average volume of lymphocytes, granulocytes and monocytes based on histograms of the distribution of cells by volume issued by the device. To assess the statistical significance of the differences in variables, the authors used the Mann–Whitney U-test.&#x0D; Results. In the experimental group of animals after 30 days of ore intoxication, the average volume and number of granulocytes and monocytes increased statistically compared to the control. The authors have revealed a correlation between the parameters of cell distribution and the average volume in leukocyte populations at different times of ore intoxication.&#x0D; Conclusion. The effect of copper-zinc pyrite ore on the condition of the leukocyte link of peripheral blood was an increase in the average population volumes of all types of leukocytes by the 30th day of the experiment, due to a change in the ratio of the number of cells of different sizes and the degree of heterogeneity of the populations.&#x0D; Limitations. The limitation of this study is the model of oral administration of copper-zinc pyrite ore in the form of an aqueous suspension.&#x0D; Ethics. The authors have conducted the study in compliance with the "Rules of work using experimental animals". The study was approved by the local Ethics Committee of the Bashkir State Medical University, Ministry of Health of the Russian Federation, Protocol No. 5 of 13.09.2017.

Immunomodulatory Effects of Microcin C7 in Cyclophosphamide-Induced Immunosuppressed Mice.

Microcin C7 (McC) as a viable immunomodulator peptide can be a potential solution for pathogenic microbial infection in the post-antibiotic era and has gained substantial attention. This study was designed to evaluate the immunomodulatory activity of Microcin C7 in a cyclophosphamide (CTX)-induced immunodeficient mouse model. We show that Microcin C7 treatment significantly alleviated the CTX-caused body weight loss, improved the feed and water consumption to improve the state of the mice, and elevated the absolute number and proportion of peripheral blood lymphocytes as well as the level of hemoglobulin. We further aim to characterize the phenotypes of the immune function and intestinal health profiles. The results demonstrate that Microcin C7 treatment increased serum levels of immunoglobulin A (IgA), IgG, interleukin 6, and hemolysin, promoted splenic lymphocyte proliferation induced by concanavalin A and LPS, and enhanced the phagocytosis of peritoneal macrophages immunized by sheep red blood cells. Additionally, Microcin C7 treatment decreased levels of diamine oxidase and d-lactate, ameliorated CTX-induced intestinal morphological damage, and increased the levels of zonula occluden 1, occludin, claudin-1, mucin 2, and secretary IgA in the jejunum and colon. Moreover, Microcin C7 administration is sufficient to reverse CTX-induced intestinal microbiota dysbiosis by increasing the number of Lactobacillus and Bifidobacterium, decreasing the number of Escherichia coli in colonic contents. Collectively, our results demonstrate that Microcin C7 may have protective and immunomodulatory functions and could be a potential candidate used in animal feed, functional foods, and immunological regimens..