Activity Of Blood Neutrophils Research Articles

Release of metal ions from dental alloys and their effect on the metabolic and killing activity of leukocytes

Aim of study. We investigated the effect of metal cations solutions from dental alloys and solutions of metal salts on the functional and metabolic activity of blood leukocytes. Materials and methods. 22 patients with complaints of intolerance to dental materials (IDM) were examined. The control group consisted of 19 patients without IDM complaints. Results. After the incubation of fragments of dental alloys in water, metal cations are identified, depending on the time of incubation. This metal cations solutions (MCS) changed the metabolic and killing activity of blood neutrophils in patients with IDM, which may be a marker of intolerance to prosthetic materials. MCS and metal salts solutions (MSS) increased the secretion of IL-2, IL-4, TNF-α, INF-γ by whole blood leucocytes in patients with IDM, which indicates that hypersensitivity to the components of dental alloys was induced in the experimental group, and only IL-2, IL-10 were upregulated in the control group, which in turn indicates an immunomodulatory effect. Secretion of cytokines by blood leukocytes of patients influenced by components of dental alloys can serve as a marker of their biocompatibility and a criterion for hypersensitivity reactions. The serum level of HSP-70 in patients with IDM was 2.41 [0.79; 2.9] ng / ml, and it was lower in comparison with the control group– 4.1 [2.23; 5.11] (p <0.05). The PHA stimulated the release of HSP-70 in both groups. In patients of the experimental group MCS inhibited the secretion of HSP-70 (0.8 [0.3, 1.1] ng / ml), while in the control group it stimulated its secretion (5.7 [4.1, 8.5] ng / ml). The baseline NO-producing activity of leukocytes in patients of both groups during incubation with PHA in vitro did not differ. However, when they were cultivated with MCS, the release of NO by blood leucocytes was suppressed.

EFFECT OF LIPOPOLYSACCHARIDE OF ESCHERICHIA COLI ON PHAGOCYTE AND METABOLIC ACTIVITY OF MICE BLOOD NEUTROPHILS WITH INDUCED IMMUNE DEFICIENCY

Aim. Experimental evaluation of effect of E. coli LPS fractions on phagocyte and metabolic activity of blood neutrophils of laboratory mice with induced immune deficiency. Materials and methods. Phagocyte activity was evaluated by phagocyte number (PN), phagocyte index (PI) and integral phagocyte index (IPI), intensity of metabolism and energetic processes of enzyme systems - by test of tetrazolium nitro blue (TNB), mean cytochemical coefficient (MCC) in spontaneous and induced tests and stimulation index (SI). Results. LPS-3 substance administration into mice with secondary immune deficiency has resulted in a significantly highest increase of PN (15.8%), IPI (17.7%), TNB-IN (10.3%), cytochemical coefficients MCC-IN (u.) and IS (u.) - 14.8 and 10.9%, respectively, compared with the parameters of immune deficient mice that had received licopid. Conclusion. Some fractions of E. coli M17 polysaccharide have immune stimulating activity.

Relationship between immune and oxidant disorders
 associated with acute non-obstructive pyelonephritis

Acute pyelonephritis is the most common disease of urinary system in all age groups resulting from an infectious-inflammatory process leading to mobilization of the immune system. A key event is the contact of the pathogen with the cells of the immune system that form the first line of defense and are capable of phagocytosis and destruction of microbial agents that cause oxidative disorders. The aim of the study was to establish the relationship between the immune and oxidant disorders at the systemic and local level in acute obstructive and non-obstructive pyelonephritis. 56 patients with acute serous and serous non-obstructive acute obstructive pyelonephritis confirmed by instrumental methods were under constant observation. Plasma and urine concentrations of cytokines (TNFα, IL-1β, IL-6, IL-8, IFNα, IFNγ, IL-4, IL-10, IL-1RA), components of the complement system (C3, C3a, C4, C5 and C5a, Factor H, C1-inhibitor) were determined along with activity of neutrophils in peripheral blood, concentration of peroxidation products (malonic dialdehyde, acylhydroperoxide), catalase and superoxide dismutase activities. The level of immune inflammation and oxidative stress were higher in acute obstructive pyelonephritis than in acute non-obstructive pyelonephritis. The close correlation between the studied oxidant and immune parameters at the systemic and local levels can be used to assess the severity of the disease, its prognosis, treatment and preventive measures.

Fluctuation in the number, type and activity of blood neutrophils in cows exhibiting successful and unsuccessful completion of gestation cycle

Neutrophils (first line of cellular defense) are capable of detecting presence of foreign genome in the mother’s womb. Role of neutrophils during full gestation cycle of ruminants and the difference in their number, type, and activity in successful and unsuccessful pregnancies is not known. To evaluate this, blood samples were collected at artificial insemination (0 day) and on days 10, 14, 16, 18, and 21 in non-pregnant (NP) cows. However in pregnant (P) cows, samples were collected as indicated above and every 30 days for the complete gestation. In aborted cows, samples were collected as above till abortion. Higher total leukocyte counts were observed in NP and aborted cows at abortion. Neutrophil: lymphocyte ratio increased significantly (p < 0.05) in NP and aborted cows. Phagocytic activity (PA) and myeloperoxidase concentrations were significantly higher (p < 0.05) on day 18 post insemination in NP cows. PA and myeloperoxidase also increased significantly (p < 0.05) at abortion in aborted cows. Neutrophils exhibited limited decrease in their number and activity in successful pregnancies during implantation. After that their number and activity were constantly maintained throughout the gestation cycle. Any increase in the number and inflammatory activity of neutrophils may lead to non-pregnancy or loss in pregnancy.

The phagocytic activity of blood neutrophils in patients with chronic generalized periodontitis of I-II severity level on the background of parasitosis

The article presents the results of a study of phagocytic immunity chain in patients with CGP of I-II severity level on the background of enterobiasis, toxocariasis, giardiasis. The aim of this work was to study the nature of changes in the phagocytic immunity chain in patients with CGP of I-II severity level with parasitoses and patients with CGP without parasitoses. Studies were conducted on 540 patients with CGP, I-II severity level with parasitosis - (principle groups). The comparison group consisted of 90 CGP patients, I-II severity level without parasitosis, the control group – of 30 people without periodontal diseases and chronic diseases of other organs and systems. Phagocytic activity of neutrophils was assessed by their ability to absorb the S.aureus bacteria (strain 209), bactericidal capacity (BC) of phagocytes by the S. Nielsen method. Oxygen-dependent activity of phagocytes was studied in the spontaneous and induced zymosan HCT-test. Statistical processing of materials was made using mathematical statistical methods for the data analysis. There was established a reduction in patients with CGP of I-II severity level with parasitosis and in patients with CGP without parasitosis of the absorption and biocidal ability of peripheral blood neutrophils compared with healthy individuals. At I degree of CGP severity in patients without parasitosis, the number of cells, involved in phagocytosis (FI) was decreased on 13.7%, the number of microbes, absorbed by one cell (FN) on 17.3%, biocide – on 76%. As follows from the obtained data FI and FN in this group of patients did not differ from normal values significantly. In patients with CGP of I-II severity level without parasitosis all three indicators studied had a statistically significant difference. FI was reduced in comparison with the norm on 20.5%, FN – on 24.2%, BC – on 152%. In patients with CGP of I-II severity level with parasitosis the significant differences from the control group were observed both at the I degree of the disease, and at II severity level of the disease. At I degree of severity of the disease, reduction of FI in patients with parasitosis was 22.2 28.0%, FN – 32.8 – 39.7%, BC 158 – 170%, at II degree FI – 29.3 – 34.4 %, FF – 41.4 – 53.5%, BC – 238 – 264%. In patients with CGP of I-II severity level with parasitosis the significant differences compared with patients with CGP of I-II severity level without parasitosis were respectively related to the number of absorbed microbes by a single cell (FN) and biocidal capability of neutrophils (BC). Phagocytic index (PI) of patients with CGP of I-II severity level with parasitosis did not differ significantly from those in patients with CGP of I-II severity level respectively without parasitosis. In patients with CGP of I-II severity level with various forms of parasitosis is reduced as the induced red HCT test, as well as spontaneous HCT test. In patients with CGP of I-II severity level without parasitic, the spontaneous HCT test is increased in comparison with both infested patients, and the control group of individuals.

Peripheral blood neutrophil activity profiles in dogs with spinal cord injuries: a longitudinal analysis

A challenge in treating secondary spinal cord injury (SCI) has been developing agents that can penetrate the central nervous system. Thus, there is an emphasis on targeting peripheral immune responses post‐injury, as it is well recognized that a majority of inflammatory cells within the injured cord originate from the periphery. Limited data in rodents and human SCI shows an increase in circulating neutrophils, enhanced neutrophil oxidative burst activity, and an increase in circulating monocytes. This study was carried out in dogs with naturally‐occurring SCI resulting from intervertebral disk herniation (IVDH) to longitudinally characterize the peripheral post‐injury immune cell populations. These studies are feasible since we can obtain serial blood draws, and can measure clinical outcomes. Texas A&amp;M see approximately 120 cases of canine IVDH associated SCI each year, which are primarily dachshunds breed. This data from dogs can be extremely useful since canine SCI parallels human SCI with respect to identifiable contusion with sustained compression; treatment modalities including decompression and physical rehabilitation; and histopathic, molecular and magnetic resonance lesion phenotype. The objective of our laboratory is to provide a large animal model of naturally‐occurring SCI to recapitulate translational rodent research, where blood and cerebrospinal fluid volumes are too low to analyze longitudinally. This project focuses on identifying and enumerating resting and activated neutrophils in dogs with SCI, peripheral injury (long bone fracture), and in healthy controls recruited from Texas A&amp;M University. Blood draws of 10 mL were collected on the day of admission and 3, 7, 30 and 90 days post‐SCI, and on single day from dogs with long bone fractures and healthy controls. The modified Frankel score (MFS) was used as a coarse measure of injury severity to stratify patients into severe vs. mild‐moderate injury. Isolated neutrophils from peripheral blood were evaluated using commercially available canine and mouse antibodies: CADO48A (canine specific neutrophil marker), CD11b, CD45, and CD62L (L‐ selectin). Neutrophil oxidative burst activity was determined by observing the conversion of non‐fluorescent dihydrorhodamine123 (DHR) to fluorescent rhodamine123 by cellular oxidation. To our knowledge, this is the first time longitudinal analysis of peripheral neutrophil activation has been performed in dogs. Preliminary results from a small number of animals show that there are differences in L‐selectin expression and mean fluorescent intensities of DHR conversion detected after SCI or peripheral injury compared to healthy controls. The increased numbers of CADO48Ahigh/CD62Lhigh cells in SCI dogs compared to peripheral injured dogs could demonstrate these cells as targets for immune therapy, and provide a window for therapeutic intervention. This study demonstrates changes in the peripheral immune cells in an animal model that closely resembles human pathogenesis of neuroinflammation after spinal cord injury.Support or Funding InformationSupported by the Helen McWhorter Chair at Texas A&amp;M University

The respiratory explosion activity of neutrophils in blood from patients with Graves' disease manifestation

&#x0D; Objective. To study respiratory burst activity in neutrophilic granulocytes in the onset of Graves’ disease (GD).&#x0D; Material and methods. Twenty-six females aged 18—55 years (mean age, 40.7±13.2 years), with the diagnosis of Graves’ disease verified for the first time, were enrolled in this study. Spontaneous and zymosan-induced chemiluminescence (CL) was assessed using a CL3606 36-channel analyzer for 90 min. The following parameters of the chemiluminescence curve were determined: time to attain the maximum (Tmax), the maximum intensity (Imax), and the area under the curve (S).&#x0D; Results. Patients with GD and lucigenin-dependent CL of neutrophils exhibited decreased zymosan-induced CL and the increased Imax of luminol-dependent spontaneous and zymosan-induced CL of neutrophils. Association between the anti-TPO level in blood serum with Tmax (r=–0,70; р=0.036) and Imax of luminol-dependent induced CL (r=–0.72; р=0.030) was found. The maximum level of synthesis of secondary reactive oxygen species (ROS) was found to be elevated in patients with GP both at relative rest and upon antigen-induced respiratory burst.&#x0D; Conclusion. In patients with GD, respiratory burst activity in neutrophils increases due to synthesis of secondary reactive oxygen species. Upon the onset of GD, already at the stage of subclinical thyrotoxicosis, the association between the indices of respiratory burst in neutrophils and thyroid hormones is lost but there emerges a relationship between the CL response and anti-TPO concentration.

Titanium dioxide stimulates phagocytic activity of blood neutrophils

The aim. Evaluation of the TiO2 effect on the phagocytic activity of peripheral blood neutrophils. Materials and methods. 17 people were examined - 10 people with atopic bronchial asthma (m/f - 5/5, mean age - 38 [28, 48]), verified according to the international criteria, the second group (control) - 7 people - healthy controls (m/f - 3/4, mean age - 23 [21; 26] years). The phagocytic index and neutrophil activity in the NBT-test were calculated for all the examined patients. 0.008% solution of TiO2 (based on 1 mg / kg of body weight) was added to the test-samples, control samples were added with the physiological solution, standard samples - with a suspension of killed Staphylococcus aureus. All the samples were incubated for 30 minutes at 37°C, the ready smears were microscopied with counting the amount of formazan-positive granulocytes in the NST-test and cells that absorbed 1 or more Stf particles per 100 granulocytes. Results. TiO2 stimulated phagocytosis of staphylococcus with blood neutrophils in 53% of the examined patients by 8-30% (p = 0.02). Under the influence of TiO2 activity of phagocytes significantly increased, in comparison with that in spontaneous NST-test (p = 0.0001), but was lower than induction by staphylococcus (p = 0.049). In patients with allergopathology, TiO2 stimulated phagocytosis of staphylococcus and neutrophil activity in the NBT-test was also effective, as was the culture of staphylococcus aureus (31.2% compared to 34.3%). In the healthy group, the TiO2 solution increased the activity of neutrophils in the NST-test, compared with the physiological solution (on average and in 71% of cases by 10-18%), but not so than the culture of staphylococcus (24%, compared to 39, 7%). Among healthy people, there was a correlation between the hereditary of allergy and the neutrophil activity in the NBT-test with TiO2 (0.78). Conclusions. It was found that 0.008% solution of TiO2 stimulated the activity of neutrophils in the NBT-test, both in patients with atopic bronchial asthma and healthy people after 30 minutes of incubation in a thermostat. In patients with allergies, TiO2 increased phagocytosis of staphylococcus, as well as metabolic activity of neutrophils so much as staphylococcus. A strong correlation was found between the hereditary allergies and the increased metabolic activity of blood phagocytes in the NBT-test with TiO2 in healthy individuals.

Influence of the Probiotic Prolam TM at the Oxidase Activity of Blood Neutrophils of Newborn Calves

The aim of our research was to study the influence of different doses of the probiotic Prolam TM о n the germicidal activity of neutrophils of calves’ blood in a newborn period. Our studies have shown that the daily watering of probiotic Prolam TM to the calves within 10 days after birth in a dose of 5 ml did not have a material impact on the adaptive reserve of neutrophils having oxidase activity . The use of this probiotic in a dose of 20 ml optimized homeostasis that was evidenced by the reduction of the relative content of neutrophils in the blood of calves to the normative values and caused the rising of the oxygen - dependent germicidal activi ty of neutrophils of calves blood. The selection of effective doses of probiotics can be justified indicators oxidase activity of blood neutrophils of newborn calves.

Absorption Activity of Blood Neutrophils of Calves in Different using Schemes of Protamine

Absorption Activity of Blood Neutrophils of Calves in Different using Schemes of Protamine

INFLAMMATORY BOWEL DISEASE: CYTOCHEMICAL ACTIVITY OF INTRACELLULAR ENZYMES OF PERIPHERAL BLOOD NEUTROPHILS AND MONOCYTES IN DIAGNOSIS

Aim. To carry out literature analysis devoted to the studies of the importance of peripheral blood neutrophils and monocytes in the pathogenesis and diagnosis of inflammatory bowel diseases, namely Crohn's disease and ulcerative colitis. Blood neutrophils and monocytes, as the main components of the inflammatory infiltration of the intestinal wall, are the sources of inflammatory mediators, pro-inflammatory cytokines, involved in the mechanisms of the mucous membrane destruction. Change of these blood cells intracellular enzymes activity using cytochemical methods can indicate the activity of the systemic inflammatory process, which reflects the activity of this process in the intestinal wall, and make possible prognosis of the disease course. Conclusion. Further study of the dynamics of the functional activity of blood neutrophils and monocytes will make possible design of diagnostic algorithms for inflammatory bowel diseases and assess the effectiveness of the therapy.

Combined treatment with MSC transplantation and neutrophil depletion ameliorates D-GalN/LPS-induced acute liver failure in rats

The imbalance of immunity is an important pathogenesis of acute liver failure (ALF). Neutrophils are the hallmark of acute inflammation, which have an essential role in immune regulation. Mesenchymal stem cell (MSC) transplantation is a promising therapy in ALF treatment. Recent studies indicated a considerable connection between MSCs and neutrophils in immune regulation. To investigate changes in neutrophils in ALF rats after MSC transplantation, and to explore the therapeutic effect and mechanism of the combined treatment with MSC transplantation and neutrophil depletion in ALF. We employed monotherapy and the combination therapy with MSCs and anti-PMN serum in D-galactosamine (D-GalN)/lipopolysaccharides (LPS)-induced ALF rats. Rats were sacrificed at 6, 12 and 24h, respectively. Blood samples and liver tissues were collected. Hepatic injury, inflammatory cytokines (TNF-α, IL-1β and IL-10), chemokines (CXCL1 and CXCL2), the number and activity of neutrophils and animal survival were assessed at fixed times. MSC transplantation can effectively improve the liver function of ALF rats and reduce the number and activity of neutrophils in both peripheral blood and liver. Compared with MSC transplantation alone, anti-PMN treatment and co-treatment had a better result in diminishing neutrophils. The co-treatment also exhibited a better therapeutical effect in ALF rats compared with monotherapy. In this process, the expressions of inflammatory cytokines in the liver were consistent with liver function. The regulation of the neutrophil-related microenvironment is affected in D-GalN/LPS-induced ALF rats after MSC transplantation. The combined treatment with MSC transplantation and neutrophil depletion may have a better therapeutic effect in ALF rats.

Role of neutrophils in treatment of rats with D-galactosamine-induced acute liver failure with bone marrow mesenchymal stem cells

Objective: To investigate the therapeutic effect of bone marrow mesenchymal stromal cell (BMSC) transplantation on D-galactosamine-induced acute liver failure in Sprague-Dawley (SD) rats, as well as the mechanism of neutrophils in this process. Methods: A total of 39 male SD rats were divided into control group (8 rats, intraperitoneal injection of isotonic saline), model group (10 rats, intraperitoneal injection of D-galactosamine), solvent group (9 rats, tail vein injection of isotonic saline at 2 hours after intraperitoneal injection of D-galactosamine), and treatment group (12 rats, tail vein injection of MSCs at 2 hours after intraperitoneal injection of D-galactosamine). The rats were sacrificed at 24 hours after the model of D-galactosamine-induced acute liver failure was established, and the blood and liver tissue were harvested. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and total bilirubin (TBil) were measured, and blood analysis was performed to measure the number and percentage of neutrophils in peripheral blood. Immunofluorescence assay was used to measure the expression of the neutrophil marker Ly6g in the liver, the myeloperoxidase (MPO) kit was used to measure the activity of MPO in liver, and RT-PCR was used to measure the mRNA expression of inflammatory cytokines and chemokines in the liver, i.e., tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interferon-γ(IFN-γ), interleukin-10 (IL-10), CXC chemokine ligand 1 (CXCL1), and CXC chemokine ligand 2 (CXCL2). Another 64 male SD rats were randomly divided into groups, and the survival rates of rats in each group were observed for 7 days. The independent samples t-test was used for comparison between any two groups (Levene homogeneity test of variance, and the corrected t-test was used for a P value of < 0.05), and the log-rank test was used for comparison of survival rates between any two groups. Results: At 24 hours after acute liver failure was induced by D-galactosamine in the SD rats, there were significant increases in the liver function parameters (ALT: 2884.1±541.0 U/L vs 45.4±11.0 U/L,P< 0.001; AST: 3634.9±755.9 U/L vs 143.9±23.7 U/L,P< 0.001; TBil: 44.4±8.4μmmol/L vs 0.9±0.2μmmol/L,P< 0.001) and the number and percentage of peripheral blood neutrophils [number: (4.7±1.1)×109 vs (1.4±0.4)×109,P< 0.001; percentage: 44.9%±8.0% vs 18.3%±4.4%,P< 0.001]. A large number of neutrophils aggregated in the liver tissue, and there were significant increases in the MPO activity (4.72±1.09 U/g vs 1.13±0.24 U/g,P< 0.001), inflammatory cytokines, and chemokines. Compared with the model group, the treatment group showed significant improvements in liver function (ALT: 1 823.9±389.2 U/L vs 2 884.1±541.0 U/L,P< 0.001; AST: 2173.0±567.3 U/L vs 3634.9±755.9 U/L,P< 0.001; TBil: 30.9±6.5μmmol/L vs 44.4±8.4μmmol/L,P< 0.001) and survival rate (50% vs 12.5%,P= 0.023). Meanwhile, the treatment group also showed significant reductions in the number and percentage of peripheral blood neutrophils [number: (3.5±1.0)×109 vs (4.7±1.1)×109,P= 0.012; percentage: 35.9%±8.9% vs 44.9%±8.0%,P= 0.021], number of neutrophils in the liver, and MPO activity (3.52±1.03 U/g vs 4.72±1.09 U/g,P= 0.040), as well as significantly inhibited expression of inflammatory cytokines and chemokines (TNF-α: 2.458±0.762 vs 3.778±1.046, P = 0.005; IL-1β: 2.498±0.547 vs 4.065 ± 0.953,P= 0.002; IFN-γ: 3.977±1.039 vs 5.418±1.255, P = 0.025; IL-10: 6.056±1.542 vs 3.368±0.952,P= 0.001; CXCL1: 7.988±1.911 vs 10.366±1.239,P= 0.010; CXCL2: 3.441±1.005 vs 4.847±1.113,P= 0.019). Conclusion: BMSC transplantation has a therapeutic effect on D-galactosamine-induced acute liver failure in rats, and this process is accompanied by reduced aggregation and activity of neutrophils in peripheral blood and liver. Inflammatory cytokines and chemokines may be involved in the mechanism of regulation of these two aspects.

Фагоцитарная активность нейтрофилов крови у пациентов с хроническим генерализованным пародонтитом I-II степени тяжести на фоне паразитозов

Фагоцитарная активность нейтрофилов крови у пациентов с хроническим генерализованным пародонтитом I-II степени тяжести на фоне паразитозов

Effects of supplementation of polyherbal-potash alum mixture on immune status of crossbred cows during transition period

The present study was conducted to investigate efficacy of supplementation of polyherbal-potash alum mixture on immune status of cows during transition period. Fourteen crossbred cows in their late gestation were selected and randomly divided into 2 groups. Cows in one group were supplemented with mixture containing herbs and potash alum from 30 days ante-partum to 30 days postpartum. Trachyspermum ammi Asparagus racemosus, Ocimum sanctum, Emblica officinalis, Cinnamomum zeylancium and potash alum were used for preparation of mixture. Immune status viz, total leucocyte counts (TLC), differential leucocyte counts (DLC), in-vitro phagocytic activity (PA) of blood neutrophils and plasma cortisol level were estimated as well as incidence of diseases were studied. Results showed that supplementation mixture significantly (P&lt;0.05) reduced cortisol level and TLC, whereas phagocytic activity was increased on the day of parturition. However, no significant (P&lt;0.05) changes were observed in DLC. Incidence of mastitis, metritis and retention of foetal membrane (RFM) in control group was 28.57%, 28.57% and 14.28%, respectively whereas no such cases were found in treatment group. The study clearly indicated that supplementation of polyherbal-potash alum mixture enhance immunity during transition period.

Effects of Different Dietary Selenium Sources on Antioxidant Status and Blood Phagocytic Activity in Sheep.

The objective of this experiment was to investigate the effects of feed supplementation with equivalent doses of selenium from sodium selenite (SS) or selenized yeast (SY) on Se deposition, selenoenzyme activity and lipid peroxidation in tissues as well as in bacterial and protozoal fractions of rumen contents in sheep. The phagocytic activity of monocytes and neutrophils in whole blood was also assessed after 3months of dietary treatment. While animals in the control group were fed with unsupplemented basal diet (BD) containing only background Se (0.16mg/kg DM), the diet of the other two groups (n=6) consisted of identical BD enriched with 0.4mg Se/kg DM either from SS or SY. Concentrations of Se in blood and tissues were found to be significantly increased in both supplemented groups. No response in Se deposition was recorded in the musculus longissimus dorsi of sheep given dietary SS. The intake of SY resulted in a significantly higher Se level in the blood, kidney medulla, skeletal muscles, heart, intestinal and ruminal mucosa than in the case of SS supplementation. No differences appeared between tissue Se contents in the liver and kidney cortex due to the source of added Se. Regardless of source, Se supplementation to feeds significantly increased the glutathione peroxidase (GPx) activity in blood and tissues except the kidney medulla and jejunal mucosa. Supplementation with SY resulted in significantly higher activity of thioredoxin reductase in the liver and ileal mucosa, and also reduced malondialdehyde content in the liver and duodenal mucosa. Dietary Se intake increased Se concentrations in the total rumen contents and bacterial and protozoal fractions. The accumulation of Se in rumen microbiota was associated with increased GPx activity. Phagocytic cell activity was enhanced by Se supplementation. Our results indicate that Se from both sources has beneficial effects on antioxidant status in sheep and can be utilized by rumen microflora.

EFFECTIVENESS OF APPLICATION OF COMBINED THERAPY OF EARLY ERYTHEMA STAGE OF LYME DISEASE TAKING INTO CONSIDERATION IMMUNOGENESIS FEATURES

Study innate and adaptive immunity in patients with migrating erythema, clinical effectiveness ofcombined therapy using Immunovac vaccine and dynamics of immunologic parame- ters as a result of the therapy. 37 adult patients with migrating erythema were examined. The patients were divided into 2 groups: 1st gr. (14 individuals) - Immunovac by intranasal-subcutaneous method against the background of basic therapy; 2nd gr. (23 individuals) - 200 mg/day doxycycline therapy for 21 days. Phagocytic activity of blood neutrophils; TLRs expression on mononuclear leukocytes of peripheral blood (PBML) and skin cells in foci by flow cytometry with mAT against TLR2, 3, 4, 5, 6, 7, 8, 9 using flow cytometer FC-500; subpopulation composition of peripheral blood lymphocytes; levels of pro-, anti-inflammation and regulatory cytokines in blood sera by EIA method; IgG, 1gM, IgA in blood sera were studied in patients before treatment and 1 month after therapy. A high level of TLR2, 4, 7, 8 on skin cells in foci, TLR2, 4 - on blood cells; low content of CD95+ and CD25+, high-level of serum IL-lb, IL-2 and IL-4, an increase of general IgE level was detected in patients'with migrating erythema. Immunovac facilitated an increase of CD95+ and CD25+, IFN-y synthesis, reduced the level of general IgE in a more pronounced way than basic therapy. Inclusion of Immunovac into therapy of patients With migrating erythema facilitates increase of clinical effectiveness and correlates with correction of immunologic disorders.

Prevention of mixed respiratory infections in young cattle

The possibility of use of phytomedicine in conjunction with hyperimmune serum of animal donors to enhance the preventive effectiveness at mixed respiratory infections in calves was studied. In a farm in Nizhny Novgorod region with unfavorable in terms of mixed respiratory infections of viral and bacterial etiology a high level of morbidity (80%) and mortality (26%) was revealed. Before use of medicines in clinically healthy calves aged 20-30 days mild immunodeficiency was found characterizing by decrease in relative T-cell content by 20.0%, functional activity of blood neutrophils in spontaneous and induced tests by 19.0%, lysozyme activity by 29.0% when compared with minimum value of the physiological norm for this age period. On the background of the immunological imbalance drastically decreases the effectiveness of traditional therapies. Use of Maxophyt in conjunction with hyperimmune serum of donor animals promoted statistically significant rise in initially reduced immunological parameters and intense specific immunity in clinically healthy calves: absolute and relative T- and B-cell content increased by 36.0%, 38.0% and 50.0%, 67.0% respectively, functional activity of blood neutrophils in spontaneous and induced tests by 18.0% and 32.0%, IgG and IgM levels by 46.0% and 53.0% respectively, lysozyme activity of blood serum by 60.0%. Subcutenous injection of hyperimmune serum of donor animals in conjunction with aerosol treatment with phytomedicine Maxophyt enhanced the preventive effectiveness at mixed respiratory infections of viral and bacterial etiology to 96.2% which was 21.2% higher when compared with the control and average daily body weight gain to 890.0 ± 22.8 g which was 26.0% higher when compared with the control. Combination of medicines provided the 100% safety of calves in experimental groups which was 8.3% higher when compared with the control. The course of disease in experimental calves was mild.

Infradian rhythm in hematology, cell mediated immunity and serum micronutrient concentration around peripartum periods in high yielding crossbred cows before and after micronutrient supplementation

Alteration in hematology and cell-mediated immunity around peripartum periods before and after micronutrient supplementation was studied in this investigation. Twelve (12) high-yielding cross-bred cows in advanced pregnancy were selected for the experiment and divided into two equal groups viz. supplemented (n = 6) and unsupplemented (n = 6). Supplemented animals were provided with a micronutrient mixture (@ 25 g per cow) as per the recommendation of NRC over the normal feeding. Unsupplemented group of cows were kept as control with standard farm feeding management. Blood samples (15 ml/cow) were collected from all the animals during days 30th, 15th, and 7th before calving, on the day of calving, and during days 7th, 15th, and 30th after calving and analyzed for hematology viz. hemoglobin (Hb), total erythrocyte counts (TEC), total leukocyte counts, and differential leukocyte counts. In vitro phagocytic index (PI) of neutrophils and lymphocyte proliferation response (LPR) were also evaluated. Copper (Cu), zinc (Zn), manganese, and selenium (Se) concentrations in serum were estimated by atomic absorption spectrophotometer. The Hb concentration was found to be significantly (p < 0.01) higher in supplemented group compared to the unsupplemented group of cows. There were no significant differences in Hb concentration during different days of peripartum in both the group of cows. There was a significantly (p < 0.001) higher TEC in supplemented group compared to unsupplemented group of cows, though it did not vary significantly between different days of peripartum in both the group of cows. There was a significantly (p < 0.001) higher PI in supplemented group compared to the unsupplemented group of cows. In both the groups, the PI of blood neutrophils was significantly (p < 0.001) lower during calving. The LPR did not differ significantly between the groups, but it differed significantly (p < 0.01) between different days of peripartum and was lowest on the day of calving in both the groups. The Cu, Zn, and SE concentration was found to be significantly (p < 0.01) higher in supplemented group compared to the unsupplemented group of cows, but remained unaltered throughout the peripartum period. The study indicated that there was a marked immune depression around peripartum (15 days precalving to 15 days postcalving) in terms of in vitro phagocytic activity of blood neutrophils and in vitro LPR. Supplementation of micronutrients improves the hematological status as well as immune status, so it was recommended to supplement these micronutrients for better productivity and disease resistance in high-yielding cross-bred cows around peripartum.

Comparative Efficiency and Impact on the Activity of Blood Neutrophils Isolated by Percoll, Ficoll and Spontaneous Sedimentation Methods

ABSTRACTStudies on the role of cells in physiological and pathological processes generally require isolation of some populations, such as neutrophils. In the literature, several methods used for isolating neutrophils are described; however, there is no consensus on the best technique to be used in cell functional studies. The present study compares the efficiency and impact on the chemotactic and phagocytic activity of neutrophils isolated from blood by three different methods: Percoll and Ficoll density centrifugation gradients and spontaneous sedimentation technique. The neutrophil chemotaxis, stimulated with lipopolysaccharide (LPS), autologous serum or homologous serum, was determined by using Boyden chambers. The phagocytic capacity was assessed by ingestion of zimosan particles, and digestion phase was analyzed by nitroblue tetrazolium test (NBT). The results obtained from neutrophil isolation by Percoll and Ficoll density gradients, as compared to spontaneous sedimentation technique, showed similar degrees of cell yields and higher purity; however, these methods affected neutrophil responsiveness, accompanied by elevated chemotaxis and reduced chemotactic capacity to respond to subsequent stimulation. Neutrophil isolation by spontaneous sedimentation, in contrast, did not affect cellular activity and resulted in cell preparation with high number of neutrophils. Although neutrophil phagocytosis results were similar between the different methods, digestion phase of phagocytosis was significantly enhanced after LPS-stimulation, only in the neutrophils isolated by spontaneous sedimentation technique. In conclusion, the present study shows that isolation of blood neutrophils by the spontaneous sedimentation technique is appropriate for the assessment of cellular activity, since it neither primes or activates the neutrophils nor does it affect their functional responsiveness.