Total Number Of Leukocytes Research Articles

Overexpression of Homeobox A1 Relieves Ovalbumin-Induced Asthma in Mice and Is Associated with Blocking of the NF-κB Signaling Pathway.

Homeobox A1 (HOXA1) is a protein coding gene involved in regulating immunity signaling. This study aims to explore the function and mechanism of HOXA1 in asthma. An asthma mouse model was established via ovalbumin (OVA) induction. Airway hyperresponsiveness was evaluated by the value of pause enhancement (Penh). Inflammatory cells in bronchoalveolar lavage fluid (BALF) were detected by Trypan blue and Wright staining. The pathological morphology of lung tissues was assessed by H&E staining. The IgE and inflammatory biomarkers (IL-1β, IL-6, IL-17, and TNF-α) in BALF and lung tissues were measured by ELISA. Western blot was performed to detect the expression of NF-κB pathway-related proteins. HOXA1 was down-regulated in OVA-induced asthmatic mice. Overexpression of HOXA1 decreased Penh and relieved pathological injury of lung tissues in OVA-induced mice. Overexpression of HOXA1 also reduced the numbers of total cells, leukocytes, eosinophils, neutrophils, macrophages, and lymphocytes, as well as the levels of IgE, IL-1β, IL-6, IL-17, and TNF-α in BALF of OVA-induced mice. The inflammatory biomarkers were also decreased in lung tissues by HOXA1 overexpression. In addition, HOXA1 overexpression blocked the NF-κB signaling pathway in OVA-induced mice. Overexpression of HOXA1 relieved OVA-induced asthma in female mice, which is associated with the blocking of the NF-κB signaling pathway.

CLINICAL AND IMMUNOLOGICAL CHARACTERISTICS OF PATIENTS WITH ACUTE CORONARY SYNDROME WHO UNDERWENT COVID-19

This study examined the difference between neutrophil phagocytosis and humoral immunity in patients with acute coronary syndrome (ACS) who had Covid-19 compared with those without a new coronavirus infection. We examined 65 men with ACS aged 40 to 65 who had Covid-19 within 6 months to one year prior to the study, and 20 patients with ACS who did not have a new coronavirus infection and were vaccinated against coronavirus. All patients had critical stenoses of the coronary arteries and underwent stenting of these vessels in the first 3 days from the moment of admission to the hospital. In the group of patients with ACS who did not have covid (vaccinated), persons with unstable angina prevailed, while in the group with Covid-19, patients with acute myocardial infarction (AMI) prevailed, and to a greater extent with ST elevation (nST), which is a more severe form of ACS. All deceased patients with ACS had Covid-19. It was found that in patients who underwent Covid-19, Jg M (p <0.0001) and Jg G (p <0.01) to the virus significantly predominated in comparison with those who did not have a new coronavirus infection (vaccinated). Patients who underwent Covid-19 and ACS (p<0.05) showed a higher duration of hospitalization, blood troponin level, almost 3 times, and a Grace risk, an increased number of stents, and complications — stent thrombosis. They also showed a tendency to increase the total number of leukocytes, neutrophils, neutrophil-lymphocyte ratio in comparison with patients who did not have a new coronavirus infection. Higher levels of monocytes were observed both in absolute and relative values, at the same time lower values of activity and intensity of phagocytosis and phagocytic number of neutrophils were determined in people with ACS and Covid-19 compared to those without a new coronavirus infection. In the group of patients with ACS who underwent Covid-19, higher levels of general and specific Ig M, G to Covid-19 were noted.

Цитологические особенности мононуклеарных лейкоцитов, культивированных в присутствии полиаминов

Quantitative and qualitative changes in the cellular composition after incubation with putrescine and cadaverine are considered. The aim of the study was to study changes in the cytological characteristics of leukocytes after their incubation with cadaverine and putrescine. The number, cell types, sizes, presence of vacuoles and inclusions in peripheral blood leukocytes (n=15) were evaluated after their preincubation with cadaverine (5 and 25 mmol/L) and putrescine (5 and 25 mmol/L) for 72 hours at 37oC. It has been shown that as the concentration of putrescine increases, the total number of leukocytes decreases, while cadaverine dose-dependently increased this indicator. At the same time, after exposure to putrescine, the proportion of large lymphocytes and the number of vacuoles in the cytoplasm increased. Cadaverine at a concentration of 25 mmol/L significantly reduced the number of cells with signs of cytolysis. Thus, in the course of the conducted studies, it was found that cadaverine and putrescine, produced mainly by prokaryotic cells, affect both the number and cytological characteristics of immunocompetent cells.

APPLICATION OF VERMIGHUMUS IN FEEDING PIGS

The article analyzes a new method of fattening young pigs by introducing a biologically active additive into the diet - vermighumus, which is a product of pig manure processing by California worms. In addition to the standard diet, the youngsters of the research groups consumed vermighumus in a dose of 130, 180, 230 g per head per day. The young of the control group consumed standard compound feed. It was established that the application of vermighumus into the diet of young pigs improved their fattening indicators. In particular, the youngsters of the second and third experimental groups exceeded the control analogues in terms of live weight at the end of fattening by 3.73 and 5.53%, respectively. Youngsters of the second and third experimental groups also had higher growth energy by 4.80 and 7.40%, respectively, and better feed payment (by 0.43 and 071 kg). In terms of half-carcass length, young animals of the experimental group 3 did not significantly exceed the counterparts of the control group. In the 2nd and 3rd experimental groups, the fat thickness at the level of the 6th-7th thoracic vertebra was smaller (by 5.85-7.50%). They prevailed over the control analogues in the area of the "muscle eye" (by 2.83-5.45%). The weight of the rear third half of the carcass and the content of meat in the carcass was the highest in young animals of the 3 experimental group (by 7.85% and 4.91%, respectively), and the fat content in it was lower than in the control group (by 14.52%). The smallest fat thickness at the level of the 6-7 thoracic vertebra, as well as the content of fat in the carcass, was in young animals of the third experimental group. Among the indicators of the physical and chemical properties of the muscle tissue of young pigs of the experimental groups, no statistically significant differences were found. However, there is a certain trend towards a decrease in such indicators as the level of acidity, tenderness and losses during cooking, and an increase in the moisture-holding capacity and intensity of coloring in the young of the control group. The tasting evaluation revealed minor differences among the experimental animals. The relative number of lymphocytes (% of the total number of leukocytes) in pigs treated with vermighumus was higher (by 4.44-6.66% Р<0.05) than in counterparts fed with conventional compound feed, which indicates an improvement in resistance animalsEnrichment of the ration of young pigs with vermihumus as a biologically active additive contributed to the improvement of fattening and meat qualities and ensures obtaining additional basic products in the amount of UAH 320.38-354.34 per head.

PS-BPB04-8: ANGIOTENSIN II FORMULATIONS AND THEIR IMPACT ON CARDAIC, VASCULAR AND INFLAMMATORY MECHANISMS OF HYPERTENSION

Angiotensin II (AngII) is a major effector of the renin-angiotensin system, with a critical role in BP regulation of vascular function, and end-organ damage. AngII infusion, using osmotic minipumps, is the most common animal model of hypertension, recapitulating key vascular and cardiac features of human hypertension. While different formulations of AngII are available, their impact on the key outcomes of the model remains unknown. Accordingly, we compared two most commonly used AngII formulations, and their impact on the key cardiovascular endpoints in a mouse model of hypertension. Male 12-week old C57BL/6J mice (n = 11–14) underwent Sham, AngII[Val5] or AngII[Ile5] (490 ng/min/kg) treatment for two weeks, using osmotic minipumps. Blood pressure was measured by tail-cuff, vascular function by wire myography, remodelling by picrosirius red staining, mRNA expression using real-time PCR and perivascular (PVAT) inflammation by flow cytometry. Data were analyzed using one-way or RM-two-way ANOVA. Both AngII formulations caused a significant BP elevation (p < 0.001) in comparison to sham animals, although AngII[Val5] had a more pronounced pressure effect than AngII[Ile5] (171 ± 4 vs 156 ± 6 mmHg, p = 0.03). Similarly, endothelium-dependent relaxation of resistance arteries and aorta was significantly impaired by AngII infusion (p < 0.001), however, a maximum Ach-induced relaxation was significantly lower in AngII[Val5] than in AngII[Ile5]. Both, AngII formulations increased intima-media thickness of the aorta (p < 0.0001), although the effect of AngII[Val5] was ∼2-fold higher than AngII[Ile5] (p = 0.001). While animals infused with AngII[Val5] had enhanced vascular collagen deposition (p < 0.001), this effect was lesser in the AngII[Ile5] group (p = 0.02). The total number of leukocytes infiltrating PVAT was induced by both AngII formulations, however, this effect was less pronounced in AngII[Ile5] (1002 ± 376 vs. 753 ± 242 cell/mg, p = 0.03). The same effect was found among CD11b+ and CD11c+ cells. While a significant increase of CD3+ cells was observed in both AngII formulations. While cardiac hypertrophy was evident in both AngII formulations this effect was significantly lower in AngII[Ile5] (p = 0.03). This was confirmed by cross-sectional area of cardiomyocytes. Histological analysis of heart sections showed a significant effect of both AngII formulation on cardiac fibrosis (p < 0.05). This effect was more pronounced upon AngII[Val5] infusion (p < 0.001) and was associated with higher expression of pro-fibrotic markers. AngII[Val5] formulation is a more efficient and consistent way to study key cardiovascular endpoints such as BP, inflammation, vascular and cardiac damage compared with AngII[Ile5]. Furthermore, the use of AngII[Val5] leads to a more pronounced phenotype and may be associated with reducing the number of animals needed in experiments.

INFLUENCE OF CYTRIMIN ON LEUKOCYTE FORMULA STRUCTURE OF FISH

An obvious trend in domestic and foreign aquaculture in recent years is usage of biologically active substances of various mechanisms of action. It was clearly demonstrated that such biologically active substances as adaptogens, probiotics, vitamins, amino acids have a positive effect on the fish organism, increasing economic parameters of the industry. In particular, usage of "Irkutin" adaptogen in early ontogenesis of fish increased survivability of larvae during transition to exogenous nutrition, survivability of the fry, growth rate, endurance and it also reduced the level of cannibalism and stress. Therefore, the search for new substances with the properties of adaptogens for fish farming is of great importance. There is reason to believe that "cytrimin" can become such new substance. "Cytrimin" is a complex of tris-(2-hydroxyethyl) amine with zinc bis-(2-methylphenoxyacetate) and has the formula: (HOCH2CH2)3N Zn(OOCCH2OC6H4CH3-2)2. A study of the spectrum of physiological activity of "cytrimin" (one of the representatives of protatrans) showed that it protects blood vessels from formation of cholesterol plaques, it is also an antidote to alcohol and can increase performance, claiming to be an adaptogen. So far, "cytrimin" has not been used in fish farming. The aim of the work was to study the effect of "cytrimin" on parameters of blood system, in particular, on the structure of leukocyte formula of the African sharptooth catfish. It was found that there were no significant changes in the content of the total number of leukocytes in the blood of African catfish in case of "cytrimin" application, however, a redistribution of functionally different groups of cells in the leukogram of the African catfish was revealed. This redistribution was expressed in a slight decrease in the proportion of lymphocytes with a significant increase in the proportion of segmented neutrophils and monocytes within the physiological norm.

Hematological and immune disorders in colorectal cancer patients with liver metastases after radiofrequency ablation

Background. Colorectal cancer (CRC) is one of the most common malignant neoplasms in the world. It ranks third in the structure of oncological morbidity and second in the structure of mortality. The liver is the most common site of metastatic spread of CR and 14 to 18% of patients have liver metastases at diagnosis. Radiofrequency ablation is currently the most promising method of treating metastatic lesions.
 Рurpose – to study quantitative changes in hematoimmunological indicators and their influence on antitumor reactivity in patients with colorectal cancer after RFA of liver metastases.
 Materials and Methods. Clinical and laboratory examination was carried out in 12 patients with colorectal cancer with metastases in the liver, the majority of patients were over 60 years old. Adenocarcinoma was histologically determined in all patients, and most of them had a moderate degree of malignancy (G2). The study was carried out in three stages: I – one day before radiofrequency ablation (RFA), II – 3 days after RFA, III – 14 days after RFA of liver metastases.
 Results. Immune and hematological indicators of the development of the inflammatory response after RFA in patients with colorectal cancer with liver metastases were established. On the 3rd day (II stage) after RFA, a significant increase in the total number of leukocytes, the ratio of neutrophils to lymphocytes, the number of eosinophils, a violation of the balance of CD4+ and CD8+ lymphocytes, a decrease in the number of NK- and NKT-cells, an increase in the percentage of CD3+ HLA-Dr -, CD4 +PD1+, CD8+PD1+ lymphocytes. On the 14th day (stage III), most of the parameters were close to those determined before the treatment, except for the reduced number of NK and NKT cells and the increase in the level of platelets.
 Conclusions. A number of hematological changes on the 3rd day after RFA related to the inflammatory reaction were identified: an increase in the total number of leukocytes, an increase in the ratio of neutrophils/lymphocytes from 1.72 to 4.12, a significant decrease in the relative and absolute number of eosinophils. But on the 14th day, after the inflammatory reaction subsided, the normalization of most of the studied indicators was observed. A violation of the subpopulation composition of lymphocytes was established in patients with CR metastases in the liver, on the 3rd day after RFA. Those that had taken place even before the intervention (low number of CD8+ lymphocytes and NK cells) and additional ones appeared (increased CD4+/CD8+ ratio, increased percentage of CD3+ cells HLA-Dr+, CD4+ and CD8+ lymphocytes PD1+ (CD279+), decrease in the number of NKT cells). On the 14th day, the picture approached the initial one, with the exception of the number of NK and NKT cells. We believe that timely correction of inflammatory immunosuppression in the early days after RFA can shorten the period of vulnerability to recurrence of CR, and in the long term potentiate the positive effect of RFA on antitumor reactivity.

Electrocardiographic Findings and Cardiac Troponin I Assay in Dogs with SIRS Diagnosis.

Several studies performed in humans have demonstrated that the onset of systemic inflammatory response syndrome (SIRS) represents a high risk condition to develop myocardial damage and arrhythmias. Therefore, we also hypothesized cardiac involment for dogs affected by SIRS. To assess this hypothesis, 24 dogs with a diagnosis of SIRS (13 entire males, 7 entire females, and 4 spayed females) with an age ranging from 4 to 11 years (mean 5.6 years) and an average weight of 24 kg (range from 5 to 47 kg) were enrolled. The dogs were divided into two groups according to their prognosis: Survivors (G1) and not survivors (G2), composed by 13 and 11 dogs, respectively. Moreover, healthy dogs were included as the control group (CTR). All the dogs with a history of cardiac or renal disease were excluded. At the inclusion, each patient underwent a physical examination and a complete cell count, and a biochemistry panel (including electrolyte profile) was performed; moreover, the blood cardiac Troponin I (cTnI) was measured. For each clinical variable indicative of SIRS, a score between 0 (absence) and 1 (presence) was applied. Furthermore, an electrocardiographic examination was recorded. Seventeen out of 24 (70.8%) dogs with SIRS showed arrhythmias, of which n. 6 belonged to the G1, while n. 11 belonged to the G2. Most represented findings were sinus tachycardia (7/17; 41.1%), followed by monomorphic premature ventricular beats (6/17; 35.3%), less common were first-degree atrioventricular block (2/17; 11.7%) and sinus bradycardia 1/17; 5.8%). Notably, in G1 dogs, only sinus tachycardia and premature ventricular beats were observed. G2 dogs presented a number of total and banded leukocytes significantly higher than those of G1 (p = 0.002 and 0.049), in the same manner, the clinical score suggestive of SIRS (3 vs. 2.1) was significantly higher in G2 than in G1 dogs (p = 0.01). Moreover, a significantly higher value of cTnI was observed in the G2 group compared to the G1 group (p = 0.006). Data presented here suggested a cardiac involvement in dogs with SIRS, analogously to humans, that may significantly influence the patient's prognosis.

1,2,3,4,6-O-Pentagalloylglucose Protects against Acute Lung Injury by Activating the AMPK/PI3K/Akt/Nrf2 Pathway.

An acute lung injury (ALI) is a serious lung disease with a high mortality rate, warranting the development of novel therapies. Previously, we reported that 1,2,3,4,6-O-pentagalloylglucose (PGG) could afford protection against ALI, however, the PGG-mediated protective effects remain elusive. Herein, PGG (60 and 30 mg/kg) markedly inhibited the lung wet/drug weight ratio and attenuated histological changes in the lungs (p < 0.05). A pretreatment with PGG (60 and 30 mg/kg) reduced the number of total leukocytes and the production of pro-inflammatory cytokines IL-6 and IL-1β in bronchoalveolar lavage fluid (p < 0.05). In addition, PGG (60 and 30 mg/kg) also attenuated oxidative stress by reducing the formation of formation and the depletion of superoxide dismutase to treat an ALI (p < 0.05). To further explore the PGG-induced mechanism against an ALI, we screened the PGG pathway using immunohistochemical analysis, immunofluorescence assays, and Western blotting (WB). WB revealed that the expression levels of adenosine monophosphate-activated protein kinase phosphorylation (p-AMPK), phosphoinositide 3-kinase (PI3K), protein kinase B phosphorylation (P-Akt), and nuclear factor erythroid 2-related factor (Nrf2) were significantly higher in the PGG group (60 and 30 mg/kg) than in the lipopolysaccharide group (p < 0.05); these findings were confirmed by the immunohistochemical and immunofluorescence results. Accordingly, PGG could be effective against an ALI by inhibiting inflammation and oxidative stress via AMPK/PI3K/Akt/Nrf2 signaling, allowing for the potential development of this as a natural drug against an ALI.

Absence of CCR2 Promotes Proliferation of Alveolar Macrophages That Control Lung Inflammation in Acute Respiratory Distress Syndrome in Mice.

Acute respiratory distress syndrome (ARDS) consists of uncontrolled inflammation that causes hypoxemia and reduced lung compliance. Since it is a complex process, not all details have been elucidated yet. In a well-controlled experimental murine model of lipopolysaccharide (LPS)-induced ARDS, the activity and viability of macrophages and neutrophils dictate the beginning and end phases of lung inflammation. C-C chemokine receptor type 2 (CCR2) is a critical chemokine receptor that mediates monocyte/macrophage activation and recruitment to the tissues. Here, we used CCR2-deficient mice to explore mechanisms that control lung inflammation in LPS-induced ARDS. CCR2-/- mice presented higher total numbers of pulmonary leukocytes at the peak of inflammation as compared to CCR2+/+ mice, mainly by enhanced influx of neutrophils, whereas we observed two to six-fold lower monocyte or interstitial macrophage numbers in the CCR2-/-. Nevertheless, the time needed to control the inflammation was comparable between CCR2+/+ and CCR2-/-. Interestingly, CCR2-/- mice presented higher numbers and increased proliferative rates of alveolar macrophages from day 3, with a more pronounced M2 profile, associated with transforming growth factor (TGF)-β and C-C chemokine ligand (CCL)22 production, decreased inducible nitric oxide synthase (Nos2), interleukin (IL)-1β and IL-12b mRNA expression and increased mannose receptor type 1 (Mrc1) mRNA and CD206 protein expression. Depletion of alveolar macrophages significantly delayed recovery from the inflammatory insult. Thus, our work shows that the lower number of infiltrating monocytes in CCR2-/- is partially compensated by increased proliferation of resident alveolar macrophages during the inflammation control of experimental ARDS.

Maternal Breastfeeding Methods, Breastmilk-Derived Antibodies and Cells Concentrations and Their Impact on Infant Morbidity: Results from a Prospective Cohort in Southern Benin

Breastfeeding is associated with long-term well-being including low risks for infectious diseases and non-infectious diseases such as asthma, cancer, autoimmune diseases and obesity during childhood. However, very few studies assessed the real-impact of different methods of breastfeeding on infant morbidity in low-income countries such as Benin. Our study aimed to evaluate the dynamic change of immune components and the impact on infant morbidity among Beninese children population. Seventy-six children and their mothers were enrolled in colostrum, transitional and mature milk groups at the District Hospital of Comè in Bénin. Breast milk samples were collected one-time from mothers to assess total IgA, IgG and IgM and leukocytes by using spectrophotometry and level of microscopy, respectively. Mean or proportion comparisons were appreciated using a Mann-Whitney or Fisher’s exact tests, when appropriate. Forty (54.63%), six (7.89%) and thirty (39.47%) mother-infant pairs were enrolled in colostrum, transitional and mature milk groups respectively. The total number of leucocytes and antibodies were different in colostrum, transitional and mature milk. The prevalence of breastfeeding was 90.79% (n=69) in the population with 69.74% (n=53) of exclusive breastfed. Forty-five infants (84.91%) among exclusively breastfed infants were healthy whereas 4 (25%) among mixed breastfed infants and 4 (57.14%) among formula group were healthy (p=0.001). The total leucocytes count and IgA, IgG and IgM concentration decreases from colostrum through transition milk to mature milk. Our data showed a prevalence of exclusive breastfeeding which is associated to a positive clinical outcome on infant’s health. We have also confirmed decrease of antibodies and leucocytes during the maturation of breast milk.

Local immune state in the women with miscarriage

Functional state of immunity provides maintenance of immunological tolerance for allogeneic fetus, and high level of local protection against antigenic stimulation. Increased functional activity of phagocytic cells at systemic and local levels may cause impairment of complete fetoplacental complex, thus leading to abortion. The purpose of our study was to assess the state of local immunity in women with pregnancy loss.
 The study involved 174 women in their reproductive age. The first group consisted of 65 patients (37.4%) with a diagnosis of miscarriage, the history of 2 abortions over the period from conception to the 20th week of gestation. The second group included 37 patients (21.2%) with documented recurrent pregnancy loss, with a history of 3 miscarriages. The control group consisted of 72 conditionally healthy women (41.4%) who had 2 or more term pregnancies with the same partner, without a history of obstetric and gynecological complications. Their mean age was 366 years old. Total number and viability of leukocytes, the indices of functional neutrophil activity, their functional reserve and neutrophil stimulation index were determined in cervical mucosal samples over the first phase of the menstrual cycle.
 In the first phase of menstrual cycle, the women with miscarriage and recurrent pregnancy loss exhibited a statistically significant increase of phagocytosis by the neutrophils from cervical mucus as compared to the group of conditionally healthy women. Intensity of neutrophil phagocytosis in cervical mucus reached higher values in the women with recurrent pregnancy loss more often, compared to the control group. Evaluation of the functional reserve of cervical mucus neutrophils in the subjects with recurrent pregnancy loss and miscarriage showed a trend towards statistically significant differences: this parameter was higher in the patients than in control group. The ability of cervical mucosal neutrophils to produce reactive oxygen species (both spontaneous and induced) did not show statistically significant differences between the patients and controls.
 Hence, we have observed aberrant functional activity of neutrophilic granulocytes from cervical mucosa in the groups of women with 2 or more abortions, without changing ability of the cells to produce reactive oxygen species. This finding may be explained by prevalence of oxygen-independent mechanisms of intracellular killing, thus suggesting a role of neutrophils for impaired balance of immunological tolerance in pregnant women at the local level.

Relationships between the indices of innate immunity in sepsis depend on clinical outcomes

At the present time, sepsis is primarily considered a disorder of the immune system caused by dominant infection, manifesting with individual response reaction of the host organism. Hence, sepsis should not be classified as a distinct pro- or anti-inflammatory syndrome, but rather as a variable continuum of overlapping immune mechanisms. The role of innate immunity in sepsis is probably the leading one, since it exerts an early and nonspecific response to any foreign agent. The nature of appropriate relationships may determine current state of the immune system. To perform a deeper study of the anti-infectious protective mechanisms, we have evaluated the relationships between the immunological parameters of peripheral blood in patients with sepsis, depending on the outcome of the disease. Total number of leukocytes, and neutrophils was determined in whole peripheral blood of the patients with sepsis, along with calculation of leukocyte counts and numbers of neutrophil extracellular traps. Serum levels of procalcitonin, peptidyl-arginine deiminase 4, and cytokines were studied by ELISA assay. To study the relationship between the indices of innate immunity, we used correlation analysis, which was carried out by the Spearman rank criterion. When conducting the statistical analysis, strong correlations between the immunological parameters of peripheral blood in patients with sepsis were not revealed, regardless of the outcome of the disease. In cases of favorable outcome, 10 relationships were identified, with an unfavorable outcome, 7 relationships were registered. All the relationships were of medium strength. Regardless of clinical outcome of sepsis, the significant relationships were established between the number of leukocytes and the level of neutrophil extracellular traps; positive relationships were also found between pro-inflammatory cytokines IL-1 IL-6, TNF IL-18. However, these relationships were significant in cases with favorable outcome, while becoming weaker and losing their significance for the group with lethal outcomes.
 Among the revealed relationships, the most interesting finding was an association between favorable outcome of sepsis with IL-10 contents, which is an anti-inflammatory cytokine coordinating the innate immune response. In lethal outcomes, such relationship with IL-10 was not revealed. IL-10, a cytokine with anti-inflammatory properties may limit the immune response to pathogens and, thus, potentially prevent damage to the host. Therefore, the relationship between IL-10 and other immunological parameters of peripheral blood in sepsis may affect the outcome of this condition.

Characterization of the immune system of Ellegaard Göttingen Minipigs - An important large animal model in experimental medicine.

Interest in Ellegaard Göttingen Minipigs (EGMs) as a model in experimental medicine is continuously growing. The aim of this project is to increase the knowledge of the immune system of EGMs as information is still scarce. Therefore, we studied the postnatal maturation of their immune system from birth until 126 weeks of age. For the first 26 weeks of the study, animals were kept under pathogen-reduced conditions (SPF) and afterwards under conventional housing conditions. The development of the immune system was analyzed by monitoring changes in total numbers of leukocytes and lymphocytes of ten individuals and the composition of leukocyte populations by multi-color flow cytometry (FCM). We followed the presence of monocytes using monoclonal antibodies (mAbs) against CD172a+ and CD163+ and B cells based on the expression of CD79a. NK cells were distinguished as CD3-CD16+CD8α+/dim cells and further subdivided using NKp46 (CD335) expression into NKp46-, NKp46+, and NKp46high NK cells. T-cell receptor (TCR) γδ T cells were defined by the expression of TCR-γδ and different subsets were determined by their CD2 and perforin expression. TCR-αβ T cells were classified by their CD8β+ or CD4 expression. For monitoring their differentiation, expression of CD27 and perforin was investigated for CD8β++ T cells and CD8α together with CD27 for CD4+ T cells. We clearly detected a postnatal development of immune cell composition and identified phenotypes indicative of differentiation within the respective leukocyte subsets. Examination of the development of the antigen-specific immune system after transfer to different distinct housing conditions and after vaccination against common porcine pathogens such as porcine circovirus 2 (PCV2) revealed a markedly increased presence of more differentiated CD8+ and CD4+ T cells with central and effector memory T-cell phenotypes. To complement the findings, a PCV2 vaccine-specific antigen was used for in vitro restimulation experiments. We demonstrated antigen-specific proliferation of CD4+CD8α+CD27+ central and CD4+CD8α+CD27- effector memory T cells as well as antigen-specific production of TNF-α and IFN-γ. This study of postnatal immune development defines basic cellular immune parameters of EGMs and represents an important milestone for the use of EGMs for immunological questions in experimental medicine.

Monocytosis in the acute phase of SARS-CoV-2 infection predicts the presence of anosognosia for cognitive deficits in the chronic phase

Reduced awareness of neuropsychological disorders (i.e., anosognosia) is a striking symptom of post-COVID-19 condition. Some leukocyte markers in the acute phase may predict the presence of anosognosia in the chronic phase, but they have not yet been identified. This study aimed to determine whether patients with anosognosia for their memory deficits in the chronic phase presented specific leukocyte distribution in the acute phase, and if so, whether these leukocyte levels might be predictive of anosognosia.First, we compared the acute immunological data (i.e., white blood cell differentiation count) of 20 patients who displayed anosognosia 6–9 months after being infected with SARS-CoV-2 (230.25 ± 46.65 days) versus 41 patients infected with SARS-Cov-2 who did not develop anosognosia. Second, we performed an ROC analysis to evaluate the predictive value of the leukocyte markers that emerged from this comparison.Blood circulating monocytes (%) in the acute phase of SARS-CoV-2 infection were associated with long-term post-COVID-19 anosognosia. A monocyte percentage of 7.35% of the total number of leukocytes at admission seemed to predict the presence of chronic anosognosia 6–9 months after infection.

Leukocyte and IgM Responses to Immunization with the CIDR1α-PfEMP1 Recombinant Protein in the Wistar Rat.

The malaria vaccine is an important strategy for the global malaria elimination program, but the complexity of the Plasmodium antigen is a major hurdle in malaria vaccine development. The cysteine-rich interdomain region 1α (CIDR1α) of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is crucial in malaria pathogenesis, making it a vaccine candidate. This study investigated the leukocyte and IgM response generated after administering a CIDR1α-PfEMP1 recombinant protein injection in Wistar rats. The rats were divided into a control group, who received a physiological saline solution (PSS), and a treatment group, who were subcutaneously injected with 150 µg of purified CIDR1α-PfEMP1 protein three times at the 3-week interval. Blood samples were collected every week after each injection. The number of leukocytes were counted using a Neubauer chamber, and the IgM concentration was determined using an enzyme-linked immunosorbent assay (ELISA). Data were analyzed using an independent, paired-T test, a Mann–Whitney test, and a Wilcoxon test, based on the distribution of the data. The total number of leukocytes notably increased on day 29 (p < 0.05). The percentage of neutrophils decreased, especially on day 8 (p < 0.05), whereas the percentages of monocytes and lymphocytes increased, primarily on day 14 (p < 0.05). The IgM concentration increased on day 14 (p < 0.05). In conclusion, the CIDR1α-PfEMP1 recombinant protein may induce leukocyte and IgM responses, making it a potential malaria vaccine candidate.

Стан неспецифічної резистентності ротової порожнини у працівників промислових виробництв із шкідливими чинниками

The purpose of the study was to investigate local resistance of the oral cavity in industrial workers with harmful factors, patients with generalized periodontitis. Materials and methods. For this study, 2 examination groups were formed. The main group (61 people) includes employees of three industrial productions of the Ivano-Frankivsk region: employees of chemical production, glass production and agro-industrial complex. The control group consisted of 30 conditionally healthy individuals with intact periodontium. The age of the examinees was 25–55 years. Work experience in production is from 0.5 to 15 years. In order to study the state of local resistance in generalized periodontitis under the influence of harmful factors of the production environment, individual indicators of the immunity of the capillary blood of the gums and oral fluid were studied: the total number of leukocytes and lymphocytes; to evaluate the T-system of immunity, calculation of subpopulations of lymphocytes was performed using monoclonal antibodies CD-3, CD-4, CD-8; the level of immunoglobulins IgA, IgG and sIA; circulating immune complexes; phagocytic index, spontaneous nitroblue tetrazolium test. Results and discussion. A comparative evaluation of the results of immunological research in 61 patients with generalized chronic periodontitis of the main group and 30 healthy individuals of the control group indicated certain changes in the quantitative data of cellular and humoral factors and deviations in the functional activity of individual parameters of non-specific protection in the capillary blood of the gums of patients compared to the indicators of individuals with intact periodontium. The total number of leukocytes and lymphocytes in workers with generalized periodontitis remained within the generally accepted norm and differed insignificantly from the similar indicators of the healthy group. At the same time, a difference in the subpopulation composition of T-lymphocytes was revealed. According to absolute indicators, the number of CD4+ lymphocytes in patients with generalized periodontitis was 0.57 ± 0.04109/l, which is by 1.2 times lower compared to 0.68 ± 0.03109/l in healthy individuals. The obtained indicators of CD4+ and CD8+ lymphocytes made it possible to state a decrease in the value of the immunoregulatory index in workers with generalized periodontitis by 19.31% from the value of healthy people (1.63 ± 0.04 versus 2.02 ± 0.02), which is characteristic of immunodepressive conditions. In patients of the main group, an increase in IgA production was recorded on average to 2.05 ± 0.06 g/l, which is by 11.41% higher compared to the indicator (1.84 ± 0.02 g/l) of the control group. In workers of industries with harmful factors, with a dystrophic-inflammatory process in the periodontium, the level of average globular circulating immune complexes exceeded the value of healthy people (0.10 ± 0.03 mg/ml) by 2.5 times. This may indicate exhaustion of the clearance function of the phagocytic system and the development of hyperimmune complex syndrome. Conclusion. Having studied individual indicators of the capillary blood of the gums and oral fluid, we can conclude that in chronic exogenous intoxication with industrial xenobiotics, phenomena of acquired combined immune deficiency were observed, which were manifested by a decrease in the immuno-regulatory index CD4+/CD8+; a violation in the humoral link of immunity – activation of IgA, IgG production and a decrease in sIgA secretion, disorders of the phagocytic system, manifested by a decrease in the phagocytic activity of neutrophils and activation of oxygen-dependent metabolism (nitroblue tetrazolium test), which indicated, on the one hand, the strengthening of redox processes in these cells and a certain antibacterial potential of them, but on the other hand – the deepening of the inflammatory process of autogenous origin. Exhaustion of the body's compensatory capabilities confirmed the increase in the level of medium-globular circulating immune complexes. Therefore, significant changes in the non-specific protection of the oral cavity were observed in industrial workers suffering from generalized periodontitis

The role of cortisol and progesterone in the formation of the cellular com-position of blood in Holstein heifers

Purpose: quantitative assessment of the cell composition of the blood of the heifers of the Holstein breed and the study of the relationship between it and the level of steroid hormones - cortisol and progesterone.Materials and methods. The work was carried out in 2020-2021 on the basis of Belagash LLP (Republic of Kazakhstan), which specializes in the production of milk by using the productive potential of the Golstein breed. The diets of animal feeding were in accordance with the norms of VIZH. Heifers born in the spring of 2020 were included in the experimental group (n = 10). In its formation, the principle of close analogues was used. In the heifers of an experimental group of 3, 6, 9, 12, 15 months, blood samples were obtained by a vacuum method for conducting morphological and biochemical studies. The level of progesterone and cortisol was determined in the blood serum by the immunofortment method.Results. The total number of leukocytes in the blood of animals changed in the interval of 9.57-12.00 109/l, corresponding to the boundaries of the norm; The level of monocytes and lymphocytes fluctuated in the range of 3.33-7.80 and 58.93-75.01 %, amounting to 0.40-0.81 and 5.64-9.00 109/l. The number of red blood cells, hemoglobin and hematocrit by 15 months increased by 36.33; 12.93 and 40.18 %. At the same time, the volumetric characteristics of red blood cells (the average volume of red blood cells, the red blood cell distribution index almost did not depend on age, but their saturation with hemoglobin (the average hemoglobin content in red blood cells, the average concentration of hemoglobin in red blood cells) decreased. The level of steroid hormones – progesterone and cortisol in the blood of heifers increased with age in 46.06 and 8.24 times (p≤0.05). At the sametime, the concentration of cortisol statistically significantly correlated with the amount of red blood cells (r=0.71 - 0.95), the size of the red blood cell distribution index in volume (r = 0.74 - 0.98) and the absolute number of lymphocytes (Rr= 0.71 - 0.99). The level of progesterone, starting from 9 months of age, was interconnected with the total number of leukocytes (r = -0.63 --0.88), an absolute number of monocytes (r = 0.69 -0.89) and lymphocytes (r = -0.65 – -0.87), as well as red blood cells (r= 0.82 -0.93) and hemoglobin (r = 0.63 -0.73).

Non-invasive prognostic markers of the risk of preterm rupture of the membranes in premature pregnancy

The objective: to study the prognostic significance of non-invasive markers of endogenous intoxication for determining the risk of premature rupture of membranes (PROM) at the gestation period of 26–34 weeks.Materials and methods. During the study the patients were divided into two groups: the 1st (main) group – 80 women with PROM at 26–34 weeks of pregnancy, and the 2nd (control) group – 50 women with a physiological course of pregnancy and term delivery without complications. The mean age of the examined women was 29.60±6.30 years. To determine the diagnostic value of the level of hematological indicators (concentration of leukocytes (WBC), granulocytes (GRAN), lymphocytes (LYM) in peripheral blood) and indices of endogenous intoxication (EI) as predictors of PROM and the initiation of preterm labor at a gestation period of 26–34 weeks, ROC-analysis was performed and the average quality of these prognostic factors was established (p&lt;0.001).Results. The optimal cut-off point (cut-off value) for predicting PROM in preterm pregnancy according to the criterion of the maximum balance between sensitivity and specificity corresponded to WBC was more than 12.39/l, LYM – less than 19 %, GRAN – more than 76 %, NLR – more than 3.87, Kalf-Kalif’s leukocyte intoxication index (LII) is more than 3.42, Ostrovsky’s LII – more than 2.76. It was established that the area under the curve (AUROC) was 0.786 for WBC, 0.863 for LYM, 0.847 for GRAN, 0.866 for NLR, 0.753 for LII Ostrovsky, which testifies to the significant prognostic value of the studied predictors. Conclusions. The main non-invasive markers of the degree of activity of the inflammatory process and the severity of endogenous intoxication in premature birth with PRPO in pregnant women are: the total number of leukocytes, the level of granulocytes and lymphocytes, Kalf-Kalif LII and the neutrophil-lymphocyte ratio. In order to detect infectious markers of intra-amniotic infection and the further development of PROM and premature birth, it is recommended to calculate the level of endogenous intoxication in the second half of pregnancy using general blood analysis data on a hemolytic analyzer (WBC, GRAN, LYM, NLR).

Long-term Effects of Regular Whole Blood Donation on Peripheral Blood CD34+ Cells Population with using Leukoreduction Filters.

Background & Objective:Trapped cell population in leukoreduction filters (LRFs) contains such a significant number of CD34+ hematopoietic stem cells that can be recovered to be used in research studies. Methods:Samples (n=20) were obtained from 10 first-time donors and 10 regular blood donors with more than 30 times blood donation. After separating leukocytes from LRFs by backflushing, total leukocyte number and differential count were determined in both groups using an automated haemocytometer. Then cell viability and CD34+ cell quantification were assessed using 7- amino-actinomycin D and fluorescent-labeled monoclonal antibodies using flow cytometry, respectively.Results:Total leukocyte count was 665±164.92×106 in the first-time blood donors and 883±233.89×106 in the regular donors, which were not significantly different (P=0.08). While the number of CD34+ cells was significantly reduced in the regular donors compared to the first-time donors (0.58±0.20×106/µL vs. 0.36±0.22×106/µL; P=0.034). There was no significant difference in terms of absolute neutrophil count (10.58±3.66×06 vs. 13.17±6.45×106/µL; P=0.349), lymphocytes (7.75±3.11×106 vs. 10.38±3.77×106 /µL; P=0.917), and monocytes (2.31±0.88×106 vs. 2.59±1.09×106/µL; P=0.591) between the first-time and regular donor groups, respectively. Based on the correlation coefficients, the participants’ age had no significant effect on these variables. Conclusion:The results of this study depicted that regular blood donation reduces the number of CD34+ cells in the peripheral blood (PB) of regular donors while it has no significant effect on the ratio of myeloid to lymphoid cells of the two groups.