Venous Blood Leukocytes Research Articles

Association of gene polymorphisms with biological features of gastric cancer

Aim of the investigation. To evaluate associations of polymorphic variants of TNF (G308A), XPC (Lys939Gln), MTHFR (Ala222Val) genes with some biological features of gastric cancer. Materials and Methods. We evaluated polymorphic gene variants in 74 patients with verified gastric cancer or gastriesophageal junction cancer (Siewert III). Genotyping was performed by DNA isolation from venous blood leukocytes of the subjects followed by PCR with electrophoretic detection of the result. The association of polymorphic variants of genes with such biological features of the tumor as localization of the process, morphological type, degree of differentiation, type of tumor growth, presence or absence of lymphovascular or perineural invasion was evaluated. Results. Lys/Gln genotype of XPC gene (Lys939Gln) are 3.3 times more likely to develop gastric body cancer (p=0.04). Genotype GG of TNF gene (G308A) 5.3 times increases the development of gastric adenocarcinoma in comparison with other histological types (p=0.018), genotype GA of the same gene 5.2 times more often develop tumor with high-grade differentiation degree (p=0.032). Val/Val genotype of the MTHFR gene (Ala222Val) are significantly less likely to develop lymphatic/vascular invasion compared to carriers of the other genotype (p=0,029). Conclusion. The obtained data suggest that the genetic status of the patient itself may affect the biological characteristics of gastric cancer, which in turn may lead to a change in the patient’s treatment tactics.

Exploring the Susceptibility to Multiple Primary Tumors in Patients with Differentiated Thyroid Cancer.

It was demonstrated that differentiated thyroid cancer (DTC) patients may develop multiple primary tumors (MPT) during follow-up. Many studies showed an association between reduced telomere length and cancer phenotype; in particular, the short telomeres were associated with the development of a primary tumor. However, the role of altered telomere length in MPT development has not yet been demonstrated. The aim of this study was to evaluate the possible correlation between a short telomere length in blood leukocytes and the risk of developing MPT in DTC patients. We retrospectively evaluated 167 DTC patients followed up for a median of 13.6 years. Our control group was represented by 105 healthy subjects without any thyroid disease or present or past history of tumors. Our study groups, age-matched, were evaluated for the relative telomere length measured in leukocytes of peripheral venous blood. The relative telomere length (RTL) was significantly different in healthy subjects compared to the total group of differentiated thyroid cancer patients [p < 0.0001]. Shorter telomeres length was observed in DTC patients with (n = 32) and without (n = 135) MPT compared to healthy subjects (p < 0.0001 and p = 0.0002, respectively). At multivariate analysis, the parameters independently associated with the presence of MPT were RTL [OR: 0.466 (0.226-0.817), p = 0.018] and the familial DTC [OR: 2.949 (1.142-8.466), p = 0.032]. The results of this study suggest a role of the relative telomere length in predicting MPT development in DTC patients. Our results contribute to increasing the knowledge of the genetic mechanisms underlying MPT development in DTC patients, considering relative telomere length as a possible prognostic marker.

Adhesive Activity of Peripheral Venous Blood Leukocytes under in Vitro Conditions in Patients with Malignant Neoplasms

Цель. Установить влияние концентраций внеклеточного пула кластеров дифференциации и аутоантител на адгезивные свойства мононуклеаров периферической венозной крови людей при злокачественных новообразованиях. Материал и методы. В работе приводятся результаты иммунологического обследования онкологических больных (566) и практически здоровых лиц (103). В сыворотке крови определяли концентрации IL-1β, TNF-α, IL-6, IL-10, IFN-γ, IgA, IgM, IgG, IgЕ, аутоантител к фосфолипидам, фосфатидиловой кислоте, двухцепочечной ДНК и ревматоидного фактора. Изучали также фенотипы лимфоцитов (CD3+, CD16+, CD23+, CD25+, CD54+, CD71+) и концентрации sCD71, sCD25, sCD23, sCD54. Проведено изучение адгезивных свойств мононуклеаров in vitro лейкоцитарной взвеси венозной крови 67 онкологических больных и 23 практически здоровых лиц в возрасте 21-56 лет с параллельным иммунологическим обследованием. Результаты. Иммунные реакции на злокачественные новообразования характеризуются низкой эффективностью фагоцитов и цитотоксических эффекторных клеток при активном антителообразовании с участием IgE и значительного спектра аутоантител. Низкий уровень активности фагоцитов и цитотоксических мононуклеаров в периферической крови больных ассоциирован с подавлением их адгезии к стеклу в условиях in vitro. Снижение адгезивной активности к стеклу лейкоцитов периферической крови больных связано с подавлением способности к распластыванию, экзофагии и ядерной активности. Слабая адгезия к стеклу мононуклеаров венозной крови больных связана с повышенными концентрациями в сыворотке крови молекул адгезии (sCD54), рецепторов к трансферрину (sCD71), интерлейкину-2 (sCD25) и FcII (sCD23), которые сбрасываются клеткой при чрезмерной их концентрации на мембране. Среди адгезированных мононуклеаров больных чаще регистрируются агрегаты клеток и клазматоз, рассматриваемые как признаки неэффективности цитотоксичности. Выводы. Иммунные реакции на злокачественные новообразования характеризуются не только высоким уровнем гуморального ответа с участием IgE и значительного спектра аутоантител, но и низкой эффективностью цитотоксических эффекторных клеток и фагоцитов. Низкий уровень активности фагоцитов и цитотоксических мононуклеаров периферической крови больных взаимосвязан с подавлением их адгезии к стеклу в условиях in vitro. Снижение адгезивных свойств мононуклеаров обусловлено сбрасыванием рецепторных структур с мембраны при чрезмерной перегрузке цитозоля клетки сигналами извне. Потеря адгезивных способностей мононуклеаров сопровождается увеличением частоты формирования агрегатов и клазматоза.

TNF and MMP1 gene polymorphisms and their association with clinical aspects of colorectal cancer

Aim. To evaluate associations of polymorphic variants of some genes with such clinical aspects of colorectal cancer as gender, age of manifestation of the disease, localization of the tumor, the stage of the process at the time of manifestation. Materials and methods. Polymorphic variants of TNF and MMP1 genes were evaluated in 178 patients. The main group consisted of 110 patients with colorectal cancer, the control group consisted of 68 healthy volunteers. Genotyping was performed by DNA isolation from leukocytes of venous blood of the subjects followed by PCR with electrophoretic detection of the result. Description and comparison of differences in the frequencies of qualitative traits in independent groups was performed using Pearson’s χ2 criterion. Odds ratios (OR, 95% confidence interval) were also calculated for polymorphism variants that showed a correlation relationship. Differences at p<0.05 were considered statistically significant in the analysis. Results. Polymorphic variants of TNF gene (G308A) showed association with colorectal cancer in men and women, age of manifestation 50-59 and 60-69, 70 and older, localization of tumor in rectum and colon, I-III stages of tumor process. Polymorphic variants of MMP1 gene (1607 1G/2G) are associated with the development of colorectal cancer in women, age of manifestation 60-69 years old, colorectal cancer stage I-III. Conclusion. The results obtained suggest that not only risk factors influence the development of colorectal cancer, but also the human genetic status itself contributes to the development of tumors of these localizations.

The effect of endomorphins-1, -2 on the production of reactive oxygen species and the absorption activity of innate immunity cells &lt;i&gt;in vitro&lt;/i&gt; and &lt;i&gt;in vivo&lt;/i&gt;

Background. A high percentage of people in the world suffer from acute or chronic pain, the treatment of which is based on the introduction of morphine and its derivatives, agonists of the µ-opioid receptors, leading to the development of addiction in patients. An urgent task is to study the properties of endogenous opioid peptides, which are capable of showing not only pronounced analgesic activity, but also have a number of other effects on the periphery, while having a much smaller spectrum of side effects. Of particular interest are the immunomodulatory effects of peptide agonists of µ-receptors – endomorphins, tetrapeptides, the effects of which are not described in detail in the literature. Due to their structure and properties, endorphins can be considered as potential substitutes for low-molecular-weight opiates. The aim. To investigate the effect of endorphins-1,2 on the synthesis of oxygen radicals and the absorption activity of innate immunity cells in vitro, in vivo. Materials and methods. The object of the in vitro study is peripheral venous blood leukocytes from donors aged 22–40 years; the object of the in vivo study is peritoneal flush cells of Swiss white male mice. Oxygen-dependent microbicidal activity was evaluated using a luminol-dependent chemiluminescence reaction. Phagocytic activity was evaluated by flow cytometry. Results. It has been found that endomorphins inhibited spontaneous production of reactive oxygen species in vitro, in vivo. Endomorphin-2 reduced the intensity of respiratory explosion in stimulated cultures of peritoneal macrophages. Blockade of opiate receptors with naloxone in vitro, in vivo did not cancel the effects of endomorphins. Both peptides and naloxone had no effect on the phagocytic activity of macrophages of the peritoneal cavity of mice in vivo, but in vitro endomorphins led to an increase in the percentage of phagocytosis of peripheral blood leukocytes. Conclusions. The effects of endomorphins in vitro, in vivo are predominantly depressive.

Effect of toxic air pollutants on the cytokine release by leukocytes in patients with chronic obstructive pulmonary disease

Chronic inflammation in obstructive pulmonary disease develops in genetically predisposed individuals with prolonged or massive exposure to allergens or toxic air pollutants. This effect leads to hyperactivation of immune system and development of uncontrolled inflammatory response. The aim of the study was to determine the level of cytokines in the supernatant of leukocytes from the patients with COPD and asthma following incubation with air toxicants, i.e., a solution of cigarette smoke, an extract of cigarette tobacco, or a solution of exhaust combustion gases. The cytokines were determined by ELISA in the supernatants following exposure of peripheral venous blood leukocytes to the toxicants. To perform the assays, 10-mL samples of peripheral venous blood from the patients were taken into the test tubes with heparin (20 U/mL) in the morning time, not earlier than 2 days after therapeutic infusions of glucocorticosteroids. After gravity sedimentation, the leukocyte-rich was removed, centrifuged at 1500 rpm, then the liquid was discarded, and the leukocyte pellets were diluted with buffered saline (2 106 cells/mL). Individual leukocyte suspensions were divided into 4 wells of an immunological plate, 100 μL each. Equal volumes of test solutions simulating the effect of toxicants were added to three wells. The fourth well contained sterile isotonic sodium chloride solution (negative control). The mixtures in plates were exposed for 45 min at 37 °C followed by centrifugation for 10 min at 1500 rpm. From each well, 50 μL of the supernatant was taken and transferred to the plate for ELISA assays (under the same number). As a result, we have found that the solutions of cigarette smoke and exhaust gases caused release of IL-1β by leukocytes in the patients with asthma and COPD, but not in the samples from control group of healthy volunteers. Spontaneous increase in the IL-1β level was registered in the patients with asthma. The cigarette extracts caused an increased release of TNFα in the supernatant fluid of the patients with COPD. Upon exposure to a solution of exhaust gases, an increased level of TGF-β was revealed in patients with asthma compared to spontaneous cell activation (p &lt; 0.05), as well as an increase in IFNγ contents in the patients with COPD as compared with control group (p &lt; 0.05). No statistically significant changes were revealed for the levels of IL-6, IL-2, IL-4, IL-12, IFNα upon exposure of air toxicants on the leukocytes of patients with COPD, asthma or healthy volunteers.

The possibility of diagnosing intrauterine infection by the content of nitrite and non-thiolate nitroso compounds in maternal blood plasma.

The aim of the study is to develop a method for early diagnosis of intrauterine infection (IUI). A study of markers of inflammation in the venous blood of 60 pregnant women was conducted. The study was followed by a retrospective assessment of the outcomes of pregnancies and childbirth. Of these, 33 patients with a gestation period of more than 37 weeks (full-term pregnancy) and, accordingly, 27 patients from whom the blood sample was taken at a period of less than 37 weeks - patients with the threat of premature birth (PB). PB is the main factor contributing to the development of IUI. 27 patients were diagnosed with premature rupture of the membranes (PROM). Of these, 15 are with the threat of PB. 8 of them had a diagnosed IUI. In all cases of diagnosed PROM, including those with IUI, the concentration of nitrite and nontiolate nitroso compounds (NO2-+RNO) in the mother's blood plasma was 2.3±1.2 µM, while normally it does not exceed 0.1 µM (p<0.001). Regardless of the duration of pregnancy. The use of antibiotics in the case of PROM contributed to the normalization of the concentration (NO2-+RNO). Therefore, increasing of this indicator is result of bacterial infection. Indications of other markers of inflammation: the number of leukocytes in venous blood and in a smear of vaginal contents, the level of C-RB did not significantly change in both PROM and IUI (p>0.1). Since the concentration index (NO2-+RNO) increased in almost all cases of PREM, unlike all other clinical and biochemical indicators used in modern medicine, there is an obvious sense of its use for the current monitoring of the health of pregnant women. But it is still impossible to say unequivocally about the possibility of monitoring the fetal health by concentration (NO2-+RNO) in the mother's blood.

Characterization of polymorphisms in CFI and ARMS genes and their association with exudative age-related macular degeneration in Algerian patients.

Increasing evidence shows that polymorphisms in CFI and ARMS2 genes can influence exudative age-related macular degeneration (nAMD) risk. The aim of this study was to assess the role of CFI rs10033900 and ARMS2 rs3750846 polymorphisms in susceptibility to nAMD for the first time in the Algerian population. A total of one hundred twenty four controls and seventy two nAMD cases were included in the present study. Genomic DNA was extracted from venous blood leukocytes. CFI rs10033900 and ARMS2 rs3750846 variants were determined by using the real‑time polymerase chain reaction method. Differences in allele and genotype distribution between the cases and controls were tested with adjustment for age by logistic regression analysis. A stratification of case and control groups by age (<65 or ≥65) and by gender (male and female) was also performed. Statistical analyses were done using SPSS21.0. No statistically significant association was observed between CFI rs10033900 and ARMS2 rs3750846 polymorphisms and nAMD risk (p>0.05 for all comparisons). Stratification by age and gender did not show any significant association between these two polymorphisms and nAMD in a sample of the Algerian population. In our study, CFI rs10033900 and ARMS2 rs3750846 polymorphisms did not predispose alone to nAMD in our population. This study is a contribution to the enrichment of the bank data concerning the CFI and ARMS2 genes, reporting, for the first time, the allelic and genotypic frequencies of these genes polymorphisms characterizing the Algerian population.

Assessment of Venous Blood Leukocyte Profiles and Immunoglobulin E Levels in Atopic Dermatitis Patients.

There are differing results in the literature regarding the relationship between atopic dermatitis (AD) and peripheral blood leukocyte profiles. The aim of the study was to investigate the relationship between AD, blood immune cell profiles, and disease severity. One hundred AD patients aged 1 month to 18 years without any history of infection or systemic disease were enrolled as the study group. Serum concentration levels of total immunoglobulin E, calculated blood leukocyte profiles, and AD index severity scoring were calculated for statistical comparison. The differences between the groups for neutrophil-lymphocyte ratio and eosinophil-lymphocyte ratio were statistically different. However, there was no statistically significant correlation between the investigated parameters and AD index severity scoring. Although significant differences were found in some of the parameters between the groups, unfortunately, the lack of correlation between the parameters investigated and the disease severity index restricts their use in clinical practice.

Influence of polyamines of bacterial origin on the production of key cytokines in the culture of human mononuclear leukocytes

To date, participation of biogenic polyamines has been studied in details, with respect to regulation of microbial gene expression, interrelations between bacteria, development of their persistence state. Opportunity of their use as markers of human pathological conditions is being actively evaluated. The aim of our study was to assess the effect of bacterial diamines, i.e., cadaverine and putrescine, upon production of key cytokines (IFNγ and IL-4) in the culture of human mononuclear leukocytes. We studied leukocytes of peripheral venous blood obtained from 18 healthy male volunteers (mean age 24.0±0.6 years). The leukocytes were isolated by means of gradient centrifugation using a Ficoll-Verografin mixture. For the cultivation of lymphocytes, a micro-method and plastic round-bottom 96-well plates were used. Concanavalin A at a concentration of 5 μg/ml was used as a T cell mitogen. Polyamines were used at final concentrations of 5, 25, 50, 75 and 100 μM/L. The cultivation was carried out in humidified atmosphere with 5% CO2, at 37 °C for 72 hours. At the end of incubation, the culture medium was collected and frozen for subsequent quantitative enzyme immunoassays of cytokine concentrations (IFNγ and IL-4) (Russia). Viable cells were counted using Goryaev chamber after staining with 0.1% trypan blue solution. Statistical analysis was performed using Student’s t test or Mann–Whitney test. Addition of cadaverine at all concentrations reduced IFNγ production in the culture of mitogen-activated cells. When culturing leukocytes supplied with putrescine (5 to 50 μM/L), a dose-dependent decrease of IFNγ was observed. Upon further increase of putrescine concentrations, the IFNγ production is restored to the values of the control samples. Direct toxic effect of polyamines upon the cells was revealed. Both the diamines, at the doses of 50, 75, and 100 μM/L, caused increase of IL-4 production by the mitogen-activated cells. Such changes can be associated both with direct cytotoxic effect of cadaverine and putrescine, being mediated by changes of some metabolic pathways. In addition, the effects of polyamines upon monocytes present in culture can include their anti-inflammatory state, e.c., an increased IL-4 production. In general, cadaverine and putrescine, produced by microorganisms of various taxonomic groups, regulate the effectiveness of compensatory-adaptive reactions that ensure adaptation of microbial populations to changing or unfavorable environmental conditions.

A differential and diagnostic significance of monocytosis in treatment of moderate COVID-19 forms

Despite the relatively rare comorbidity with bacterial infections, in most cases treatment of COVID-19-associated pneumonia is accompanied by empirical antibiotic therapy. In addition, the occurrence of leukocytosis in response to glucocorticosteroid (GCS) therapy is often perceived as comorbid bacterial flora and is a reason for initiating antibiotic therapy. Therefore, an urgent task is to properly interpret leukocytosis in response to GCS therapy in COVID-19. The aim of the study was to examine dynamic changes in count of venous blood leukocytes, neutrophils and monocytes in patients with moderate COVID-19 after systemic GCS. We analyzed parameters of complete blood count in 154 patients with verified moderate COVID-19, at the Temporary Infectious Diseases Hospital, the “Patriot” Park of the Moscow Region. The comparison group (I) consisted of 128 patients without clinical signs of bacterial infection and leukocytosis observed on admission, who were prescribed GCS therapy. The control group (II) consisted of 26 subjects showing on admission signs of bacterial infection — a cough with purulent sputum combined with neutrophilic leukocytosis. The dynamics in venous blood cell count was assessed in group I of patients before the onset, 3 and 6 days after beginning GCS therapy. We also compared count of leukocytes, neutrophils and monocytes between patients with developed leukocytosis in group I vs. group II. As a result, an increased count of leukocytes, neutrophils and monocytes was revealed according to assessing complete blood count test in patients from group I on days 3 and 6 of ongoing GCS therapy. All patients with developed leukocytosis after GCS admission (103 subjects) had no clinical signs of bacterial infection. Patients with developed leukocytosis from group I had increased count of monocytes (0.90 (0.84; 1.02) on day 3 after GCS onset and 0.94 (0.87; 1.26) on day 6 of GCS) compared with group II (0.61 [0.50; 0.71]), p 0.001. The inter-group count of leukocytes and neutrophils did not differ. Monocytosis after GCS therapy may serve as a differential diagnostic criterion to distinguish between glucocorticoid-induced leukocytosis and comorbid bacterial infection. This may be one of the factors influencing a decision to prescribe antibiotic therapy.

Polymorphism of the cytochrome P450 CYP1A1 (Ile462Val) gene in the populations of Balkars and Karachays

BACKGRAUND: Cytochrome P450 is an enzyme involved in the metabolism of phase I xenobiotics, toxins, endogenous hormones and pharmaceuticals.&#x0D; AIM: Is studying the polymorphism of the CYP1A1 gene (Ile462Val, rs1048943) in the Turkic-speaking populations of the central part of the North Caucasus Region (Balkars and Karachays).&#x0D; MATERIALS AND METHODS: We analyzed DNA isolated from leukocytes of peripheral venous blood of a total of 177 unrelated Balkars and Karachays (104 Balkars, 73 Karachays).&#x0D; RESULTS: The ethnicity of the studied individuals was established, indicating their ancestors up to the third generation. The method of genotyping was performed by real-time PCR using competing TaqMan probes. As a result: the frequency of the 462Val variant in the sample of Balkars was 8,6% (95% CI 5,2113,33), slightly above the frequency range found in European populations. In the sample of Karachays, the frequency of the 462Val allele was 7,5% (95% CI 9,8213.08), which is the upper limit of values typical for European populations.&#x0D; CONCLUSION: In the Turkic-speaking populations of the central part of the North Caucasus, the CYP1A1 462Val allele occurs with a frequency characteristic of the Near Eastern and European populations.

Genetic prognostic factors for neoadjuvant chemoradiotherapy for colorectal cancer

Introduction. There has been a recent trend toward a gradual increase in the incidence of rectal cancer and a decrease in the average age of patients. These changes interpret the need to personalize treatment in each case.Objective – to evaluate the association of polymorphic variants of some genes with the results of neoadjuvant chemoradiotherapy of rectal cancer.Materials and methods. We analyzed polymorphic variants of MTHFR, XPD, XRCC1, XRCC1, P53, VEGF, EGFR, TNF, CHEK2 and MMP1 genes in 76 patients with rectal cancer who underwent preoperative chemoradiation therapy followed by surgical treatment. Genotyping was performed by DNA isolation from venous blood leukocytes of the subjects followed by polymerase chain reaction with electrophoretic detection of the result.Results. Statistical analysis of the association of polymorphic variants of the studied genes with the treatment pathomorphosis revealed significance in relation to the MMP1-1607 gene (1G &gt;2G) (p = 0.033). There was also an association of co-carrying polymorphic variants of TNF (G / A) + MMP1 (2G / 2G) genes with grade III–IV therapeutic pathomorphosis (p = 0.007).Conclusion. Carriage of recessive allele of MMP1 gene can be a predictor of favorable prognosis of preoperative chemoradiotherapy in patients with rectal cancer.

Polymorphism of CYP3A4 isoenzyme gene in patients with chronic rheumatic heart disease

AIM: This study aimed to identify the associations of CYP3A4 isoenzyme gene polymorphism with the parameters of echocardiography (EchoCG), spirometry, and endothelial function of patients with chronic rheumatic heart disease (CRHD).&#x0D; MATERIALS AND METHODS: A total of 128 patients with CRHD (15.6% men and 84.4% women) were examined. A392A, A392G, and G392G polymorphic markers were genotyped through polymerase chain reaction (PCR) with an SNP-EXPRESS electrophoretic scheme (NPF Litekh, Russia) to detect results after DNA was isolated from leukocytes in venous blood. EchoCG was implemented on an Affinity 50 apparatus (Philips, the Netherlands), endothelial function was assessed with an AngioScan01 apparatus (AngioScan-Electronics, Russia), and respiratory function was examined using a SpiroLab II spirometer (MIR Medical, Italy).&#x0D; RESULTS: The distance in a 6 min walk test did not show any significant differences among the groups: A392A 327.47 6.71 m, A392G 303.63 26.19 m, G392G 338.87 20.12 m (p=0.505). The area of the mitral opening was as follows: A392A 1.74 (1.67; 1.81) cm2, A392G 1.68 (1.45; 1.92) cm2, and G392G 1.65 (1.67; 1.81) cm2 (p = 0.214). As for the EchoCG parameters, the group of G392G homozygotes had the lowest linear dimensions of the left ventricle (the end diastolic dimension 4.83 (4.72; 4.95) cm, the end systolic dimension 2.97 (2.79; 3.14) cm, the right ventricle (2.45 [2.32; 2.58] cm), of the right atrium (4.09 [3.56; 4.62] cm), and the criteria of left ventricular hypertrophy (thickness of the interventricular septum 0.88 [0.81; 0.95] cm, and the posterior wall 0.88 (0.81; 0.95 cm). No statistically significant differences were found in the occlusion index amplitude among the groups, that is, single nucleotide replacements of CYP3A4 had no influence on the system of low-resistance arteries. Conversely, the values of the phase shift between channels (reflecting the condition of large arteries) significantly differed. The G392G polymorphism showed the worst parameters, and minimal changes were observed in the A392A group. Contour analysis demonstrated the highest augmentation index values in the G392G group, reflecting the maximal stiffness of vessels. The CYP3A4 polymorphism had no effect on the parameters of respiratory function in the studied cohort of patients. Spirometry revealed that the obstructive and restrictive parameters were not significant although homozygotes demonstrated the highest forced vital capacity of the lungs (76.5% [71.1% and 82.0%]) and forced expiratory volume for 1 s (84.6% [79.0% and 90.3%]). The maximal parameter of the vital capacity of the lungs in homozygotes for A392A (85.6% [82.3% and 88.8%]).&#x0D; CONCLUSION: Patients with CRHD homozygous for G392G had the minimum parameters of hypertrophy and dimensions of the left ventricular cavity. They also had the lowest values for the cavities of the right heart. CYP3A4 polymorphism had no effect on the parameters of respiratory function in the studied patients with CRHD.

Pterygopalatine fossa block and dexmedetomidine as components of stress-limiting anesthesia in ophthalmic surgery

BACKGROUND: To date, surgical interventions continue to be accompanied by a high proportion of complications and even death. In half of the cases, mortality and serious postoperative complications could have been prevented. After a comprehensive study of the factors affecting the prolongation of hospitalization and rehabilitation (Kehlet H., 1997; Kehlet H., Wilmore D., 2002), surgical stress response was identified as the most significant inducer of dysfunction among various organs and systems. General anesthesia, in its classical sense, is not able to provide patients with complete protection against surgical trauma. Comprehensive protection can be obtained through a combination of general and local anesthesia and stress-protective drugs.&#x0D; OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of the proposed scheme of stress-limiting anesthesia in ophthalmic surgery patients with corneal transplantation.&#x0D; MATERIALS AND METHODS: This study provides a comparative characterization of the severity of the stress response and pain syndrome in patients operated under the conditions of classical general anesthesia (group K) and those who received the scheme of stress-limiting anesthesia (group DB). Multicomponent anesthesia was administered in both groups. Sibazone was used for sedation in group K, whereas dexmedetomidine was used in group DB. The patients in group DB underwent local anesthesia before surgery. The main criteria for assessing the results were as follows: hemodynamic stability, blood glucose, level of venous blood leukocytes, and severity of pain according to the VAS in the postoperative period.&#x0D; RESULTS: Both schemes helped avoid pronounced fluctuations in hemodynamic and gas exchange parameters at all stages of the study. The analysis of blood glucose and leukocyte counts showed that patients in group K had a more significant deviation for both indicators from the preoperative level than those in group DB. In the study of pain syndrome, group K showed higher levels of pain according to the VAS than group DB.&#x0D; CONCLUSION: The scheme of stress-limiting anesthesia enables the control of postoperative pain without the additional use of narcotic analgesics and blockades in the postoperative period.

ПОЛИМОРФИЗМ TOLL-ПОДОБНЫХ РЕЦЕПТОРОВ-2 У ПАЦИЕНТОВ С ХРОНИЧЕСКОЙ РЕВМАТИЧЕСКОЙ БОЛЕЗНЬЮ СЕРДЦА

Aim. To assess the contribution of TLR2 gene polymorphism to echocardiography and spirometry parameters in patients with chronic rheumatic heart disease (CRHD). Materials and Methods. A total of 128 patients with CRHD were examined – 15.6% of men and 84.4% of women. Genotyping by polymorphic markers Arg753Arg, Arg753Gln, Gln753Gln was performed by PCR with SNP-EXPRESS electrophoretic scheme (NPF Litekh, Russia) for detection of the result after extraction of DNA from leukocytes of venous blood. Echocardiography with evaluation of linear dimensions, left ventricular hypertrophy and mitral orifice area was performed on Philips Affinity 50 apparatus; respiration was evaluated using Spirolab II spirometer. Results. The distance of the 6-minute walk test in the groups did not differ significantly: (р=0.168): Arg753Arg–314.75±6.88 m, Arg753Gln–389.69±19.92 m, as well as mitral orifice area (p=0.182): Arg753Arg–1.73(1.66;1.80) cm2 and Arg753Gln–1/70(1.61;1.79) cm2. Echocardiography showed no differences in the size of the left ventricle, but in the group of Arg753Gln heterozygotes, dilatation of the left atrium was significantly higher 5.20(5.08;5.32) cm (Arg753Arg-4.98(4.83;5.13) cm) and of the right ventricle 3.10(2.90;3.30) cm (Arg753Arg-2.66 (2.59;2.72) cm) and less pronounced values of the left ventricular hypertrophy: interventricular septum thickness 1.018(0.92;1.12) cm (Arg753Arg-1.02(0.98;1.05) cm) and of the right atrium 4.40(4.10;4.70) cm (Arg753Arg-4.54(4.33;4.74) cm). In spirometry, values of both obstructive and restrictive parameters were significantly lower in the Arg753Arg homozygous group: forced lung capacity 71.04(66.15;72.94)% (Arg753Gln-84.16(79.68;88,65)%); forced expiratory volume in 1 sec 79,05(76,87;81,23)% (Arg753Gln-88,18(84,40;91.96)%); reserve inspiratory volume 84.88(81.60;88.17)% (Arg753Gln-96.45(86.73;106.18)%); reserve expiratory volume 21.29 (18.08;24.51)% (Arg753Gln-25.93(13.93;37.93)%). Conclusion. In patients with RHD, the contribution of single TLR2 nucleotide replacements to the parameters of echocardiography and spirometry is possible, which is manifested in a decrease in external respiration function in Arg753Arg homozygotes and dilatation of the left atrium and right ventricle in Arg753Gln heterozygotes.

STRESS-LIMITING ANESTHESIA IN OPHTHALMOSURGERY

Despite the significant advances made by modern anesthesiology in the perioperative management of patients, surgical interventions are still accompanied by a high proportion of complications and even deaths. At the same time, it is known that in 50% of cases mortality and serious postoperative complications could be prevented. After performing a thorough study of the factors affecting the increase in hospitalization and postoperative recovery (Kehlet H., 1997; Kehlet H., Wilmore D., 2002), it was determined that the surgical stress response is the most significant inducer of dysfunction of various organs. and systems. To date, it has been established that general anesthesia in its classical sense does not allow achieving complete protection of the patient from surgical trauma. More complete protection can be achieved by combining general anesthesia with regional blockages and adjuvant drugs with stress-protective properties. In the present study, we compared the severity of the surgical stress response and pain syndrome in patients operated on under conditions of multicomponent balanced general anesthesia (group K) with patients who received a stress-limiting anesthesia regimen (group DB). In both groups, multicomponent anesthesia was performed, in group K, sibazone was used for sedation, in group DB, dexmedetomidine was used. Also, patients of the DB group underwent regional blockade (blockade of the pterygo-palatine fossa) before surgery. The main criteria for evaluating the results were: hemodynamic stability, blood glucose, the level of venous blood leukocytes and the severity of pain according to the VAS in the postoperative period. Both schemes made it possible to avoid pronounced fluctuations in hemodynamic and gas exchange parameters at all stages of the study. When analyzing blood glucose and leukocyte counts, it was found that patients in the control group had a more significant deviation of both indicators from the preoperative level than in the stress-limiting anesthesia group. In the study of pain syndrome, it was determined that in the control group the level of pain according to the VAS was higher at all stages of the study compared to patients in the group of stress-limiting anesthesia.

Diurnal variation of leukocyte counts affects the indirect estimation of reference intervals

Abstract It has been observed that the estimation of reference intervals of leukocytes in whole venous blood leads to higher upper reference limits (uRLs) with indirect methods than has been reported in the literature determined by direct approaches. This phenomenon was reinvestigated with a newer, more advanced indirect method, and could be confirmed. Furthermore, a diurnal variation was observed with lower values during the morning and higher values in the late afternoon and at night. This observation can explain why indirect approaches using samples collected during 24 h lead to higher uRLs than direct methods applied on samples collected presumably in the morning.

Interrelation of the content of transferrin, erythrocytes with the functional activity of leukocytes of peripheral venous blood in residents of the European North of the Russian Federation

Interrelation of the content of transferrin, erythrocytes with the functional activity of leukocytes of peripheral venous blood in residents of the European North of the Russian Federation

Дифференцировка моноцитов крови и особенности цитокинового статуса у больных туберкулезом легких

In clinical manifestation of pulmonary tuberculosis, alveolar macrophages perform as a reservoir where they accumulate mycobacteria and lose their effector functions due to the pathological conversion of macrophage pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype, which provides transition to chronic infection. The study hypothesis suggested that the cytokine status, as evaluated by leukocyte secretion of cytokines in vitro, influences the monocyte polarization in the blood during their migration to the inflammatory focus, thereby determining differentiation and pathways of macrophage activation in tissues.The aim of this work was to assess the immunophenotype of blood monocytes and the in vitro secretion of immunoregulatory cytokines by mononuclear peripheral blood leukocytes from patients with different clinical forms of pulmonary tuberculosis taking into account the pathogen sensitivity to major anti-tuberculosis drugs.Methods. 65 patients with newly diagnosed pulmonary tuberculosis were evaluated. The study material was venous blood and peripheral blood mononuclear leukocytes. Monocyte immunophenotype was determined in whole blood by flow cytometry on a Cytoflex flow cytometer (Becman Coulter, USA) with monoclonal antibodies (eBioscience, USA). Results were processed with a CytExpert 2.0 software. The number of cells expressing surface markers, CD14, CD163, CD204 and HLA-DR, was determined. Content of cytokines (IL-2, IL-10, TGFβ) in supernatants of cell cultures was measured by enzyme-linked immunosorbent assay (ELISA). Results of the study were processed with a SPSS v.11.0 standard software package.Results. The study results suggested that with an overall decrease in the number of circulating CD14-positive blood monocytes in patients with pulmonary tuberculosis regardless of its clinical form, high expression of cell activation markers remained both for the pro-inflammatory M1 phenotype (HLA-DR-positive monocytes) and the anti-inflammatory M2 phenotype (CD163-positive monocytes). In disseminated tuberculosis, the number of anti-inflammatory CD204-positive monocytes, M2 macrophage precursors, increases indicating predomination of the immunosuppressive response. In vitro analysis of the cytokine status showed that tuberculosis progression is accompanied by inhibition of effector immune responses and increases in anti-inflammatory cytokine concentrations in vitro. These changes may be equally either a cause or a consequence of deficient IL-2 secretion. We also found that the secretion of mediators with suppressor effects (IL-10, TGFβ) varied depending on both the clinical form of tuberculosis and the pathogen sensitivity to anti-TB drugs; IL-10 hypersecretion was observed in patients with drug-sensitive, infiltrative tuberculosis whereas TGFβ hypersecretion was observed in disseminated, drug-resistant tuberculosis.Conclusion. Features of blood monocyte differentiation in patients with pulmonary tuberculosis allowed us to conclude that monocytes, the macrophage precursors, start expressing markers for different functions of M1 and M2 macrophages with polarization toward the M2 immunophenotype already in the bloodstream. Therefore, in the development of pulmonary tuberculosis, cytokine regulation mechanisms become involved in suppressing the activation of innate immunity, which possibly causes chronic inflammation in the lungs and formation of Mtb-induced immunodeficiency.