CHARACTERISTICS OF CD68+ AND CD163+ EXPRESSION IN PLACENTA OF WOMEN WITH PREECLAMPSIA AND OBESITY.

OBJECTIVE: The aim: To study the peculiarities of CD68+ and CD163+ macrophage expression in the placentas of women with obesity who developed preeclampsia by applying immunohistochemical method. PATIENTS AND METHODS: Materials and methods: The study included 20 placentas taken from women who delivered full-term live-birth babies. The women were divided into 4 groups of 5 individuals each: women with physiological body weight (1st group); women with class II obesity (2nd group); women with physiological body weight and preeclampsia (3rd group); women with class II obesity, who developed preeclampsia (4th group). RESULTS: Results: The analysis of the expression level of CD68+ and CD163+ decidual macrophages shows the predominance of CD68+ pro-inflammatory profile over CD163+ anti-inflammatory profile in women of all groups. Evaluation of CD68+ and CD163+ expression levels of Kashchenko-Hofbauer cells in the stroma of the terminal villi of the placentashows that the expression level of CD68+ macrophages is significantly higher in women with obesity and preeclampsia than in the control, or in women with obesity or preeclampsia. There was a reverse tendency to the polarization shift in Kashchenko-Hoffbauer cells in the stroma of the terminal villi towards the predominance of CD163+ macrophages over CD68+ macrophages in all groups of women. CONCLUSION: Conclusions: The imbalance in anti-inflammatory and pro-inflammatory profile of placental macrophages with a predominance of the latter can lead to the development of preeclampsia.

TREATMENT OF ODONTOGENIC PHLEGMONS IN PATIENTS TAKING INTO ACCOUNT THE BIORITHM OF LIFE.

OBJECTIVE: The aim: Of our study was to establish how the biological rhythm of human affects the reparative functions of the body in terms of odontogenic purulent-inflammatory diseases of the maxillofacial localization. PATIENTS AND METHODS: Materials and methods: The research was conducted on the basis of the Department of Maxillofacial Surgery on the basis of «Poltava Regional Clinical Hospital. M.V. Sklifosovsky¼. A total of 40 patients with odontogenic phlegmons of maxillofacial localization. RESULTS: Results: On the first day of the study, the indicators of the clinical condition of patients did not have a significant difference in all study groups. On the 3rd day of all studied groups, the number of points probably decreased compared to the first day of the study by 22.5%, 23.1%, 23.7%, 22.7%, respectively. On day 5, we have observed a significant difference between the previous results in all groups: 1a - 26.6%, 1b - 23.8%, 2a - 23.9%, 2b - 24.0%. CONCLUSION: Conclusions: The most effective treatment results were observed in patients of the morning chronotype who underwent surgery in the morning. Thus, the influence of the morning chronotype of the circadian rhythm on the course of reparative processes is manifested in the later stages of reparative regeneration.

PATHOGENETIC ROLE OF MACROPHAGE COLONY-STIMULATING FACTOR (CSF-1) IN PREDICTING ENDOMETRIOID DISEASE.

OBJECTIVE: The aim: To assess the CSF - 1 level in peritoneal fluid and menstrual blood of women with endometrioid disease and to investigate its diagnostic and prognostic specificity. PATIENTS AND METHODS: Materials and methods: The study included 80 women of child-bearing age (mean age 30.95 ± 6.49 years) with benign gynaecological pathology of the ovaries and / or fallopian tubes. The women included in the study were divided into two groups: study group (n = 50, mean age 31.04 ± 6.3 years), consisting of patients with confirmed endometrioid disease, and control group (n = 30, mean age 30.8 ± 6.8 years), involving individuals without signs of endometriosis (p> 0.05). RESULTS: Results: We have found significantly higher level of CSF-1 content in the peritoneal fluid in the subjects of the study group (2027.05 ± 732.64 pg / ml) compared with those in the control group (1725.62 ± 466.06 pg / ml) (p = 0.029). There is a tendency towards an increase in CSF-1 level in women with endometriosis in its more severe stages and more severe and extended adhesions. The investigation of CSF-1 content in menstrual blood has demonstrated significant increase in its values in the women of the study group (9431.6 ± 2866.22 pg / ml) compared with the values in the control group (6637.12 ± 954.05 pg / ml), (p = 0.00004). Thus, there is a tendency towards the growth in CSF-1 level in peritoneal fluid and menstrual blood in women with endometriosis and concurrent increase in severity of the disease. CONCLUSION: Conclusions: There has been found significant increase in CSF-1 content in women with endometrioid disease in both peritoneal fluid and menstrual blood (1.2 and 1.4 times, respectively). Thus, macrophage growth factor (CSF-1) can be used as a diagnostic and prognostic criterion in evaluating the progression of endomertioid disease.

Procalcitonin in early prediction of acute severe pancreatitis.

OBJECTIVE: The aim of the study was to analyze the prognostic potential of procalcitonin in acute pancreatitis complicated by ascites-peritonitis. PATIENTS AND METHODS: Materials and methods: The study analyzed the results of a comprehensive examination and treatment of 18 patients with acute pancreatitis complicated by enzymatic ascites-peritonitis, including 13 patients who were treated in the surgical department of KP "Poltava Regional Clinical Hospital. MV Sklifosovsky POR ", and 5 patients of other emergency hospitals in Poltava, in the period from 2017 to 2019. In addition to standard screening methods, these patients were additionally tested for procalcitonin to predict an adverse course in the early period. RESULTS: Results: To assess the relationship between the presence of elevated procalcitonin levels at the time of hospitalization of 0.5 ng / ml and above and unsatisfactory treatment results, differences were assessed using an accurate Fisher test. When comparing differences in the development of infectious complications in the dynamics of the disease in patients of the study group depending on the presence of elevated concentrations of procalcitonin or its absence at the time of hospitalization, a significant difference was found (p <0.05). CONCLUSION: Conclusions: In our opinion, the use of procalcitonin as a predictor of infectious complications in the dynamics of the disease will determine the category of patients in whom reducing the risk of flora translocation through the use of early oral antibiotic prophylaxis and parenteral drugs tropic to pancreatic tissue may reduce the incidence of purulent complications. In another category of patients, antibacterial therapy is not advisable due to the low risk of purulent-septic complications.

TLR4 and TLR7 genetic polymorphism in patients with HIV/HCV-coinfection: prevalence and gender-related features.

OBJECTIVE: Introduction: HIV-infection and chronic hepatitis C is of great concern of current infectology. The paper is aimed at the study of the prevalence of the of TLR4 gene Gly and TLR7 gene Leu polymorphic alleles among patients with HIV/HCV-coinfection in general and with regard to gender, as well as to determine their role in the development of the infection. PATIENTS AND METHODS: Materials and methods: To achieve the objective of the research a cohort and case-control study has been carried out. The total of 535 people has been examined, including: HIV/HCV-coinfected - 104, HIV-monoinfected - 90, patients with chronic hepatitis C - 166 and almost healthy people (population control group) - 175 subjects. RESULTS: Results and conclusion:The study found thatthe prevalence of the TLR4 gene Gly polymorphic allele among patients with HIV/HCV-coinfection, HIV-monoinfection and chronic hepatitis C accounted for 23.1 %, 14.4 % and 14.5 %, respectively, which is significantly higher than the similar index in controls - 3.3 %. The presence of the TLR4 gene Gly polymorphic allele in the genome increases the risk of HIV/HCV-coinfection development, in case of infection, by 9 (OR=8.70, р=0.000), HIV-monoinfection and chronic hepatitis C by 5 times (OR=4.89, р=0,016 and OR=4.9, р=0.011, respectively). TLR7 gene Leu polymorphic allele is recorded with the frequency of 19.9-26.0 % in patients with HIV/HCV-coinfection, HIV-monoinfection and chronic hepatitis C as compared to controls (25.9 %). In female patients with HIV/HCV-coinfection, HIV-monoinfection, chronic hepatitis C and healthy individuals the TLR7 gene Leu polymorphic allele is recorded by 2.1-3.6 times more frequently than in male patients.

Long-Term Administration of Omeprazole-Induced Hypergastrinemia and Changed Glucose Homeostasis and Expression of Metabolism-Related Genes.

Introduction: PPIs, or proton pump inhibitors, are the most widely prescribed drugs. There is a debate regarding the relationship between long-term PPI use and the risk of type 2 diabetes mellitus (T2DM). A potential connection between T2DM and PPIs could be an elevated gastrin concentration. This study is aimed at investigating the long-term effects of PPI omeprazole (OZ) on glucose homeostasis and pancreatic gene expression profile in mice. Methods: Healthy adult male BALB/c mice were randomly divided into three equal groups (n = 10 in each one): (1) experimental mice that received OZ 20 mg/kg; (2) control mice that received 30 µl saline per os; (3) intact mice without any interventions. Mice were treated for 30 weeks. Glucose homeostasis was investigated by fasting blood glucose level, oral glucose tolerance test (GTT), insulin tolerance test (ITT), and basal insulin resistance (HOMA-IR). Serum gastrin and insulin concentration were determined by ELISA. Expressions of Sirt1, Pparg, Nfκb1 (p105), Nfe2l2, Cxcl5, Smad3, H2a.z, and H3f3b were measured by RT-PCR. Result: The ROC analysis revealed an increase in fasting blood glucose levels in OZ-treated mice in comparison with control and intact groups during the 30-week experiment. A slight but statistically significant increase in glucose tolerance and insulin sensitivity was observed in OZ-treated mice within 30 weeks of the experiment. The mice treated with OZ exhibited significant increases in serum insulin and gastrin levels, accompanied by a rise in the HOMA-IR level. These animals had a statistically significant increase in Sirt1, Pparg, and Cxcl5 mRNA expression. There were no differences in ß-cell numbers between groups. Conclusion: Long-term OZ treatment induced hypergastrin- and hyperinsulinemia and increased expression of Sirt1, Pparg, and Cxcl5 in mouse pancreatic tissues accompanied by specific changes in glucose metabolism. The mechanism of omeprazole-induced Cxcl5 mRNA expression and its association with pancreatic cancer risk should be investigated.

PPARG stimulation restored lung mRNA expression of core clock, inflammation- and metabolism-related genes disrupted by reversed feeding in male mice.

The circadian rhythm system regulates lung function as well as local and systemic inflammations. The alteration of this rhythm might be induced by a change in the eating rhythm. Peroxisome proliferator-activated receptor gamma (PPARG) is a key molecule involved in circadian rhythm regulation, lung functions, and metabolic processes. We described the effect of the PPARG agonist pioglitazone (PZ) on the diurnal mRNA expression profile of core circadian clock genes (Arntl, Clock, Nr1d1, Cry1, Cry2, Per1, and Per2) and metabolism- and inflammation-related genes (Nfe2l2, Pparg, Rela, and Cxcl5) in the male murine lung disrupted by reversed feeding (RF). In mice, RF disrupted the diurnal expression pattern of core clock genes. It decreased Nfe2l2 and Pparg and increased Rela and Cxcl5 expression in lung tissue. There were elevated levels of IL-6, TNF-alpha, total cells, macrophages, and lymphocyte counts in bronchoalveolar lavage (BAL) with a significant increase in vascular congestion and cellular infiltrates in male mouse lung tissue. Administration of PZ regained the diurnal clock gene expression, increased Nfe2l2 and Pparg expression, and reduced Rela, Cxcl5 expression and IL-6, TNF-alpha, and cellularity in BAL. PZ administration at 7 p.m. was more efficient than at 7 a.m.

Expression of periferal core molecular clock genes in oral mucosa depends on the chronotype in patients with maxillofacial cellulitis.

Introduction: Accurate determination of the patient's chronotype is one of the problems of personalized medicine. Recent studies have shown that determining of the expression of timing genes is a valuable method that can help gain molecular insight into a patient's intrinsic circadian timing. Odontogenic cellulitis is very common pathology. Since acute inflammatory diseases are an urgent pathology, the time of surgical intervention can correspond depend on the time of the patient's hospitalization. Materials and methods: The level of mRNA expression of peripheral circadian clock genes clock and bmal1, per1, cry1 in buccal epithelial cells in patients with odontogenic purulent inflammatory diseases of maxillofacial area in the morning and evening was investigated. Results: During analyzing the results of the mRNA expression study of the genes of the negative regulatory link of the peripheral molecular clock, per1 and cry1, in patients with Cellulitis of the maxillofacial area, a significant decrease (P = 0.0003) in the mRNA expression level of the cry1 gene by 2.61 times in the evening compared to its morning mRNA expression values. Conclusion: The obtained data indicate that in patients with odontogenic purulent inflammatory diseases of the maxillofacial area with an evening chronotype, a violation of the expression profile of the per1 gene in the cells of the buccal epithelium is noted, which is manifested by an increase in its evening expression in comparison with patients with a morning chronotype.

Polymorphism of tmprss2 (rs12329760) but not ace2 (rs4240157), tmprss11a (rs353163) and cd147 (rs8259) is associated with the severity of COVID-19 in the Ukrainian population.

BACKGROUND AND AIM: Angiotensin-converting enzyme 2 (ACE2), transmembrane serine 2 and serine 11A proteases (TMPRSS2, TMPRSS11A), and a cell surface cluster of differentiation 147 (CD147) might be a gene candidate that exerts the susceptibility to and mortality from coronavirus disease 19 (COVID-19). The aim of this study was to investigate the associations between ace2, tmprss2, tmprss11a, and cd147 polymorphic variants and the severity of COVID-19 in the Ukrainian population. METHODS: The study population consisted of the Ukrainian population with COVID-19: patients without oxygen therapy (n=62), with non-invasive (n=92) and invasive (n=35) oxygen therapy, as well as control subjects (n=92). Allelic polymorphisms of ace2 rs4240157, tmprss2 rs12329760, and tmprss11a rs353163 were determined by real-time PCR, and cd147 rs8259 polymorphism was detected by PCR with subsequent restrictase analysis. We compared investigated polymorphisms distribution with other populations by meta-analysis. RESULTS: Our study is the first to obtain data about the distribution of investigated gene polymorphisms in the Ukrainian population: tmprss2 rs12329760 - CC 60.9%, CT 35.9%, TT 3.2%; tmprss11a rs353163 - CC 46.7%, CT 40.2%, TT 13.1%; ace2 rs4240157 - CC 7.6%, C 18.5%, CT 22.8%, TT 19.6%, T 31.5%; cd147 rs8259 - TT 60.9%, AT 32.6%, AA 6.5%. This distribution was similar to the Northern, Western and Southern European populations. There was a statistically significant difference in the frequency of tmprss2 polymorphic genotypes CC 57.1%, CT 28.6%, and TT 14.3% (P<0.05) in COVID-19 patients with invasive oxygen therapy in comparison with non-invasive oxygen therapy. This tmprss2 mutation occurs in the scavenger receptor cysteine-rich (SRCR) domain and might be important for protein-protein interaction in a calcium-dependent manner. CONCLUSIONS: Our study indicated the presence of an association between the tmprss2 rs12329760 polymorphism and the severity of COVID-19 in the Ukrainian population.

Polymorphism of tmprss2 (rs12329760) and severity of COVID-19 in the Ukrainian population: Response.

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Interleukin-26 is associated with the level of systemic inflammation and lung functions in obese and non-obese moderate-to-severe asthmatic patients.

INTRODUCTION: Obese asthma is a complex syndrome, which includes different phenotypes of disease. At present, these phenotypes only have started to acquire a sufficient understanding. It was suggested that IL-26 is a potential biomarker of disease severity in asthma without signs of Th2-mediated inflammation. In this study, we investigated the serum and exhaled levels of IL-26 and its associations with the level of systemic inflammation, lung functions, and body weight in obese and non-obese moderate-to-severe asthmatic patients MATERIAL AND METHODS: The study included 10 healthy subjects, 10 obese subjects without lung pathologies, 10 non-obese asthmatics (NOA) (BMI 18.5-24.9 kg/m²), and 40 obese asthmatics (OA) (BMI 25.0-49.9 kg/m²). During the visit, patients' examination and spirometry with the bronchodilator reversibility test were conducted, the exhaled breath condensate (EBC) was obtained, and the blood samples were collected. The level of IL-26, interleukin-1ß (IL-1ß), interleukin-4 (IL-4), interleukin-6 (IL-6), TNF-α, interleukin-10 (IL-10), total and specific immunoglobulin E (IgE), and high sensitive C reactive protein (hs-CRP) were measured using the ELISA kits. Statistical comparison between 2 groups was analyzed using the Mann-Whitney rank-sum test. Chi-square with Yates' correction was used to compare frequencies. Spearman's rank test was used for correlating nonparametric variables. The Receiver Operating Characteristic (ROC) curve and the area under ROC curve (AUC) were used for evaluating the diagnostic power of IL-26 as a possible biomarker. RESULTS: NOA had a reversible airway obstruction with reduced FEV1, FEV1/FVC, FVC 25/75, and positive post-bronchodilator test (PBT), significantly increased serum levels of IL-10, IL-4, and slightly increased IL-26. NOA had significantly increased exhaled IL-26 in comparison with healthy subjects. The obese subjects had a normal ventilatory pattern without airway obstruction, and differences in serum IL-26, IL-10, and IL-4 concentrations in comparison with healthy subjects. Obese subjects had a significant escalation of hs-CRP and no differences in the levels of exhaled IL-26, IL-10, and hs-CRP as compared with healthy subjects. OA had reduced FEV1, FEV1/FVC, and FEV25-75 in comparison with non-obese asthmatics. OA had elevated IL-26, IL-10, IL-4, and hs-CRP concentrations as compared with healthy subjects. These patients had a partial similarity with both non-obese asthmatics (elevated IL-26, IL-10, and IL-4) and obese subjects (elevated, IL-1ß, IL-6, TNF-α, hs-CRP). OA had a reduced concentration of exhaled IL-26 in comparison with NOA and elevated exhaled IL-10 in comparison with obese subjects. Furthermore, OA had an increased concentration of IL-1ß and TNF-α in comparison with healthy individuals and NOA. Exhaled IL-26 concentration distinguished non-obese asthmatics from healthy subjects, asthmatic patients from non-asthmatics (healthy and obese subjects), all asthmatic patients from non-asthmatics (healthy and obese subjects). CONCLUSIONS: Exhaled IL-26 elevated in obese and non-obese moderate-to-severe asthmatic patients. Exhaled IL-26 might be a perspective biomarker in non-obese and obese asthmatics. The obese asthmatic phenotype comprised the combined systemic and local airway inflammation.

PPARG agonist pioglitazone influences diurnal kidney medulla mRNA expression of core clock, inflammation-, and metabolism-related genes disrupted by reverse feeding in mice.

This study examined the influence of PPARG activation by pioglitazone (PG) on the mRNA of core clock, inflammation- and metabolism-related genes in the mouse kidney medulla as well as urinary sodium/potassium excretion rhythms disrupted by reverse feeding. Mice were assigned to daytime feeding and nighttime feeding groups. PG 20 mg/kg was administered at 7 am or 7 pm. On day 8 of the feeding intervention, mice were killed at noon and midnight. Kidney medulla expression of Arntl, Clock, Nr1d1, Cry1, Cry2, Per1, Per2, Nfe2l2, Pparg, and Scnn1g was determined by qRT PCR. We measured urinary K+ , Na+ , urine volume, food, and H2 O intake. The reverse feeding uncoupled the peripheral clock gene rhythm in mouse kidney tissues. It was accompanied by a decreased expression of Nfe2l2 and Pparg as well as an increased expression of Rela and Scnn1g. These changes in gene expressions concurred with an increase in urinary Na+ , K+ , water excretion, microcirculation disorders, and cell loss, especially in distal tubules. PG induced the restoration of diurnal core clock gene expression as well as Nfe2l2, Pparg, Scnn1g mRNA, and decreased Rela expressions, stimulating Na+ reabsorption and inhibiting K+ excretion. PG intake at 7 pm was more effective than at 7 am.

Influence of age, gender characteristics, chronotype on the expression of core clock genes Per1, Clock, Bmal1 and Cry1 in buccal epithelium.

The purpose of the study is to determine the expression of the core clock genes in buccal epithelial cells of healthy people with different chronotypes. MATERIALS AND METHODS: Fourteen healthy volunteers with a healthy periodontium and oral mucosa (7 women and 7 men) were selected for participation in the trial. The buccal epithelium sampling was performed at 07:00 am and 07:00 pm in one day by cytological brush. The surveyed patients were examined chronotypically using the Horn-Ostberg test. The determination of the mRNA expression of the Per1, Clock, Bmal1, Cry1 genes was performed by quantitative real-time PCR. Statistical analysis was performed using two-way analysis of variance followed by Bonferroni post hoc tests. RESULTS: Per1 expression was higher in the morning, regardless of chronotype, age, and gender. The expression of the Clock demonstrated the prevalence of the evening in both chronotypes, in both men and women. Bmal1 was better expressed in the evening, regardless of age, gender, and chronotype. The expression of Cry1 did not show statistically significant differences between the indicators. CONCLUSIONS: The evening expression of Clock was higher in people with the evening chronotype than in people with the morning chronotype. The chronotype did not show any effect on the expression of Per1, Bmal1, and Cry1. Age and sex did not show any effect on the expression of the core clock genes.

Liraglutide exerts an anti-inflammatory action in obese patients with type 2 diabetes.

INTRODUCTION: Liraglutide (L) is the analogue of human glucagon-like peptide 1 which stimulates glucose-dependent insulin secretion and can modify the level of inflammatory biomarkers. L can influence NF-kB inflammatory cascade, but the mechanisms of anti-inflammatory activities of L remain to be determined. In animal models L influenced an activity of Sirtuin 1(SIRT1). Moreover, recent evidences strongly suggest that SIRT1 up-regulation may serve as a potent therapeutic approach against development and progression of diabetic complications. The aim of this study was to investigate L effects directed on the pro-inflammatory NF-kB pathway and expression of SIRT1 in obese patients with type 2 diabetes mellitus (DM). MATERIALS AND METHODS: 15 obese patients with type 2 diabetes were studied, all using metformin (1-2 g/day) and sulfonylurea (glimiperide). All patients received L 1.2 mg daily add-on to stable therapy for 6 weeks. Blood samples were collected before, 6 weeks after start of treatment and after an overnight fast 6 weeks after stopping L, mononuclear cells (MNC) were isolated. The mRNA expressions of TNF-α, TLR2, TLR4, NOD1, IL-2 and SIRT1 were measured in MNC by RT-PCR. Ceruloplasmin concentration was measured in plasma by photometric method. RESULTS: In this add-on pilot clinical investigation we received new data that L can inhibit proinflammatory NF-kB pathway by increased SIRT1 expression in obese patients with type 2 DM improving metabolic profile. The mRNA expression in MNC of TNF-α, IkB, TLR2, TLR4, and plasma ceruloplasmin fell after 6 weeks of L. Expressions of IL-2 and NOD-1 were stable. There was a significant increase of SIRT1 mRNA expression. The mRNA expression in MNC of TNF-α, IkB, TLR2, TLR4, NOD1, SIRT1 and ceruloplasmin concentrations did not reverse to baseline levels after 6 weeks stopping of L treatment. IL-2 expression decreased in comparison with basic level. CONCLUSIONS: L has a potent anti-inflammatory effect as do GLP-1 agonists due to inhibition of NF-kB pathways and up-regulate SIRT1 expression, down-regulating pro-inflammatory factors including cytokines (TNF-α), extra- and intracellular receptors (TLR2, TLR4), and inflammation markers such as ceruloplasmin. Long lasting effects of L can be mediated by epigenetic regulation of NF-kB pathway by SIRT-1.

Presence of Type 1 Collagen Alpha-2 (COL1A2) (rs42524) Gene Polymorphism and Scar Tissue Formation in Different Areas of Head and Neck

Abstract Objective: To determine the effect type I collagen gene polymorphism alpha-2 (COL1A2) (rs42524) on the formation of scar tissue that is localized in the head and neck areas. Material and Methods: Sixty patients with scars in different areas of the head and neck were examined. The patients were divided into four subgroups, according to the types of scarring: G I: 15 patients with normotrophic scars; G ІІ: 15 patients with atrophic scars; G ІІІ: 15 patients with hypertrophic scars; and G IV: 15 patients with keloid scars. The age of patients ranged from 17 to 54 years. The single-nucleotide polymorphic site of the COL1A2 (rs42524) gene was detected by a polymerase chain reaction and subsequent analysis of restriction fragment lengths. Pearson's chi-squared test with Yates's correction and Fischer's exact test were used. Results: There were no significant changes between the control and basic groups (p=0.83) at analyzing the frequencies of G and C alleles. For the G allele, the calculation of odds ratio between the basic and control groups was 0.93 at 95% confidence interval (CI) (0.50-1.75), for the C allele - OR was 1.07 at 95% CI (0.57-2.01). Conclusion: Our studies may indirectly indicate the activation of the skin's protective reaction to physiological scarring and dosed scar formation in different areas of the head and neck.