Total Number Of Leukocytes Research Articles

Anti-inflammatory, antinociceptive effects and involvement of opioid receptors in the antinociceptive activity of Eugenia uniflora leaves obtained with water, ethanol, and propylene glycol mixture

Ethnopharmacological relevanceEugenia uniflora (Myrtaceae) is a species native to Brazil and has a traditional use in the treatment of inflammation. Aim of the studyTo evaluate the anti-inflammatory and antinociceptive effects, and the involvement of opioid receptors in the antinociceptive activity of extract and fractions from Eugenia uniflora leaves. Materials and methodsTLC and HPLC were used to characterize the spray-dried extract (SDE) and fractions. In the in vivo assays, Swiss (Mus musculus) mice were used. Carrageenan-induced hind-paw edema and carrageenan-induced peritonitis models were used to determine the anti-inflammatory effect of the extract (50, 100, or 200 mg/kg). Acetic acid-induced writhing, tail-flick, and formalin tests were used to determine the antinociceptive effect of the extract (50, 100, or 200 mg/kg). The aqueous (AqF) and ethyl acetate (EAF) fractions (6.25, 12.5, and 25 mg/kg) were then combined with naloxone to evaluate the involvement of opioid receptors in the antinociceptive activity. ResultsIn this work, the TLC and HPLC analysis evidenced the enrichment of EAF, which higher concentration of gallic acid (5.29 ± 0.0004 %w/w), and ellagic acid (1.28 ± 0.0002 %w/w) and mainly myricitrin (8.64 ± 0.0002 %w/w). The extract decreased the number of total leukocytes and neutrophils in the peritoneal cavity (p < 0.05), at doses of 100 and 200 mg/kg and showed significant inhibition in the increase of paw edema volume (p < 0.05). The treatment per oral route (doses of 50, 100, and 200 mg/kg) significantly reduced the nociceptive response in acetic acid-induced abdominal writhing (p < 0.05). The effect of the extract on the tail-flick test showed a significant increase in latency time of animals treated at doses of 200 and 100 mg/kg (p < 0.05). The extract and ethyl acetate fraction reduced the nociceptive effect in both phases of formalin at all tested doses. The naloxone reversed the antinociceptive effect of EAF, suggesting that opioid receptors are involved in mediating the antinociceptive activity of EAF of E. uniflora in the formalin test. ConclusionThe current study demonstrates the anti-inflammatory and analgesic activities of water: ethanol: propylene glycol spray-dried extract from E. uniflora leaves using in vivo pharmacological models in mice. Our findings suggest that spray-dried extract and ethyl acetate fraction exhibit peripheral and central antinociceptive activity with the involvement of opioid receptors that may be related to the presence of flavonoids, mainly myricitrin.

Psidium guajava leaves extract decreased leukocytes and lymphocytes count in Complete Freund's Adjuvant-induced arthritis rats

Background: Psidium guajava is one of the herbal plants that has demonstrated antioxidant and anti-inflammatory properties.&#x0D; Objective: This study aimed to determine the effect of P. guajava leaves extract on leukocytes and lymphocytes in rats.&#x0D; Methods: Fifty rats were divided into five groups. Complete Freund’s Adjuvant (CFA) was employed to generate an animal model of arthritis. Group I (arthritis control) received CMC Na, group II (positive control) received dexamethasone 3 mg/kg BW, and groups III-V received 250, 500, and 750 mg/kg BW of the ethanol extract of P. guajava leaves orally. Total leukocyte counts were calculated on days 13 and 29, and on day 29, the histological joints were inspected.&#x0D; Results: The P. guajava leaves extract decreased the number of lymphocytes and total leukocytes. Before and after treatment, the group receiving doses of 250 mg/kg BW and 750 mg/kg BW of the extract showed a significant decrease in leukocytes. With treatment of 250, 500, and 750 mg/kg BW of P. guajava leaves ethanol extract, the lymphocyte count was also reduced. The P. guajava leaves extract at 750 mg/kg body weight showed the greatest effectiveness in suppressing leukocytes and lymphocytes.&#x0D; Conclusion: The extract of P. guajava leaves has the potential to be developed as an anti-inflammatory for arthritis.

Elaboration and pilot study of 3D vaccines for oncotherapy

The purpose of the study was to study the possibility of using intracorporeal constructs in vivo with healthy or autologous dead tumor cells on porous extracellular matrix of titanium nickelide (PEMTN).We aimed to determine the advantage of the effect of this method on the change in the immunological status of cancer patients in comparison with patients receiving traditional treatment.The study used CBA and AKR JY mice and studied 10 patients with solid stage IV tumors of various localization (pancreas, intestines, lungs, ovary, head and neck) and 3 oncohematological patients (lymphoblastic lymphosarcoma, chronic myeloid leukemia (CML)).PEMTN was seeded with a suspension with bone marrow cells from CBA mice and cultured for 8 weeks in a long-term Dexter culture. It was shown that after 6 weeks of incubation of bone marrow cells on PEMNT in vitro, the proportion of bone marrow cells produced from the pores of the matrix into the liquid medium varied within 58–82%, which corresponded to the figures of the 1st week of incubation.After implantation of PEMTN with fetal liver cells in AKR/JY mice with spontaneous lympholeukemia, the implant successfully engrafted and after 30 days, no noticeable fluctuations in the leukocyte count were observed. After 3–4 weeks implantation, a 3-fold increase in the level of erythrocytes with fetal hemoglobin, which is not normally found in adult AKR/JY mice, was observed. There was also an increase (by 274%, p &lt; 0.05) in the number of blood reticulocytes and a 26% decrease in the activity of the tumor process.In cancer patients with solid tumors, the level of SH-groups erythrocytes of blood increased 1.5–2.3 times 2 months after the implantation of hybrid oncotherapy implants carrying autologous apoptized tumor cells, treated in vitro. This implants, already 3–4 weeks after implantation in oncopatients, restored the blood system parameters.The most active (2–3 times) increased individual indicators of cellular immunity, in particular the number of NK cells, active (CD25+) and apoptotic lymphocytes (CD95+ marker). From the 3rd month of the study, the level of humoral immunity increased (immunoglobulins, circulating immune complexes). Clinically, the activation of immunity was accompanied by the stabilization of the process or a decrease in the volume and fragmentation of the primary nodes of solid tumors.In oncohematological patients, an increase in the CD95 marker in the peripheral cells of blood was observed.Therefore, in a patient with CML, the implantation, which followed therapy with low doses of cytostatics (myelosan, hydroxyurea) and the immune effect of the implantation, led to a 6-month hematological remission with normal values of the total number of leukocytes and leukocyte blood count. It has been shown that healthy cells in this matrix retain their viability and it allows them to multiply both in the in vitro system and in the in vivo system. The possibility of clinical use of PEMNT carrying autologous dead tumor cells treated in vitro with the aim of improving antitumor immunity, accelerating the recovery of blood counts in cancer patients and achieving partial remission has been shown.

ХАРАКТЕРИСТИКА КЛИНИКО-ГЕМАТОЛОГИЧЕСКИХ ПОКАЗАТЕЛЕЙ У ПАЦИЕНТОВ С COVID-19 АССОЦИИРОВАННОЙ ПНЕВМОНИЕЙ

Purpose of research. Assessment of the features of the clinical course of pneumonia associated with COVID-19, as well as hemogram and lymphocytic index parameters in patients with the first wave of SARS-CoV-2 coronavirus spread. Material and methods. The study included 60 patients with COVID-19 associated pneumonia (Group 1), 30 patients with bilateral community-acquired pneumonia not associated with COVID-19 (Group 2). The clinical manifestations of pneumonia and the parameters of the complete blood count were evaluated. Results. At the onset of the disease the patients of Group 1 were found to have fever (p=0.006) and dry cough (p=0.028) more often, the severity of fever being higher (p=0.019), crepitus being less frequently detected during auscultation of the lungs (p=0.03), slower regression of symptoms of pneumonia being characteristic (p=0.02) as well. Complete blood count more often determined a dysregenerative shift of the leukocyte formula to the left, manifested by an increase in the relative number of stab forms of neutrophils with a normal or reduced total number of leukocytes. As many as 45.0% patients of Group 1 showed elevated lymphocytic index (so-called excessive activation), 48.3% patients had reduced lymphocytic index, which indicates a tendency to the incompleteness of the immune responses. The inverse relationship between the lymphocytic index and the severity of pneumonia according to computed tomography (r=-0.34, p=0.0018) was established. Conclusion. Patients with COVID-19 associated pneumonia have specific clinical symptoms and hematological blood parameters as compared to those with community-acquired nonspecific pneumonia.

A Single Bout of Fatiguing Aerobic Exercise Induces Similar Pronounced Immunological Responses in Both Sexes.

Introduction: Physical exercise can acutely and chronically modulate immunological responses. Women and men have different innate and adaptive immune responses, and in this sense, these two groups may also have different acute immunological responses induced by exercise. In addition, it is essential to understand further whether the effects of physical exercise on the immune system responses depend on sex because limited scientific evidence on this topic is available. This information may allow athletes and coaches to improve the training process, mainly to understand if the physiological impact of given training stimuli in women is similar to that in men. Objective: The present study aimed to investigate the acute effects of continuous submaximal exercise until fatigue on physiological and immunological parameters in amateur female and male runners. Methods: This study included 18 female and 15 male volunteers. Each participant visited the laboratory on four consecutive days. The first visit consisted of medical history taking and explaining the study design. On the second visit, the participants were subjected to an incremental test to determine their maximal rate of oxygen consumption (VO2max) that was required to prescribe the intensity of the submaximal exercise protocol. On the third visit, the fatiguing exercise protocol was performed at 77%–80% of the VO2max. During this submaximal exercise, the heart rate, rating of perceived exertion (RPE), and blood lactate were recorded. Blood samples were collected before, immediately after, and 1 h after the fatiguing protocol to analyze the plasma levels of cytokines and creatine kinase (CK) and to count leukocytes. Finally, on the fourth visit, the participants underwent physical evaluations to measure their body composition using dual-energy X-ray absorptiometry (DXA) imaging. Results: The average ages of the female and male groups were 34.2 ± 3.7 and 30.5 ± 4.3 years old, respectively. The female group ran 57 ± 27 min, while the male group ran 52 ± 15 min before fatiguing. In the female group, when comparing before and after the submaximal exercise, marked increases were observed in the following variables: heart rate (from 68.5 to 180.4 bpm), RPE (from 3.6 to 8.2), lactate (from 2.1 to 4.49 mmol/L), and CK (from 89.5 to 126.3 U/L). In addition, the female group showed an increased number of total leukocytes (from 7222.3 to 11162.9 × 106/μl), neutrophils (from 4,403 to 6,480 × 106/μl), and lymphocytes (from 2,342 ± to 3,562 × 106/μl) from pre- to post-submaximal exercise. In the male group, similar elevations in psychophysiological variables were observed, as evidenced by comparing the heart rate (from 52.8 to 184.1 bpm), RPE (from 0.0 to 8.9), lactate (from 2.7 to 7.2 mmol/L), and CK (from 106.2 to 165 U/L) before and after the submaximal exercise. The male group also showed an augmented number of total leukocytes (from 6,245 to 8,050 × 106/μl), neutrophils (from 3,335 to 4,128 × 106/), and lymphocytes (from 2,191 to 3,212 × 106/μl) when comparing pre- and post-submaximal exercise. There were no differences in the changes between women and men for these parameters. Conclusion: The aerobically fatiguing exercise protocol induced pronounced changes in the heart rate, plasma levels of lactate and CK, total leukocyte count, especially the number of neutrophils and lymphocytes, in both sexes. The fatiguing exercise protocol also changed the plasma levels of IL-6 and IL-10 in the female and male groups. Under the present conditions, the physiological changes induced by fatiguing submaximal exercise, including the immunological changes, were not influenced by sex. This study shows that the same aerobic physical exercise can alter immunological parameters in women and men, and this response is similar between sexes.

Immunomodulatory Activity of Allium sativum, Curcuma mangga, and Acorus calamus Combination Nanoparticle on Mice Leukocytes Profile

Immunomodulators are substances acting as stimulants or suppressants against the immune system, either to specific or non-specific activity. The non-specific through the production of leukocytes as the first response in fighting against an attacking antigen. Meanwhile, the specific, hrough recognition of antigens by lymphocytes during reattack. This research aimed to determine the immunomodulatory activity of the extracts combination nanoparticles of Allium sativum, Curcuma mangga, and Acorus calamus on the leukocyte profile of mice. This study used 5 treatments and 6 repetitions. The treatment groups were K- (untreated group), P1 (extracts combination nanoparticle dose of 25 mg/kg), P2 (extracts combination nanoparticle dose of 50 mg / kg), P3 (subur kandungan herbal medicine dose of 75 mg / kg), and P4 (Clomiphene citrate dose of 0.9 mg / kgBW). The parameters used included the total number of leukocytes and their differential value. The data that met the parametric assumptions, such as normally distributed and homogeneous were examined using the One Way ANOVA test, and when there was a significant difference, it was processed with the Duncan assessment. While those that did not meet the assumptions were evaluated using a non-parametric analysis. The statistical results showed that administration of extracts combination nanoparticles of A. sativum, C. mangga, and A. calamus at doses of 25 mg/kg and 50 mg/kg were able to suppress the inflammatory reaction by decreasing the total number of leukocytes. However, the differential leukocyte count was able to maintained or modulate immune system, indicating by the percentage of neutrophil, basophil, and eosinophil in the normal range. At a dose of 50 mg/kg, decreased the percentage of lymphocytes, while for monocytes, all dosage ranges were able to increase their number.

Abstract 211: Modulating Inflammation In Atherosclerosis Through Engineered Anti-Inflammatory Cytokine Delivery

Introduction: Atherosclerosis is a chronic inflammatory disease resulting from the accumulation of low-density lipoprotein (LDL) in arterial walls. The anti-inflammatory cytokine IL-10 has been implicated in atherosclerosis with higher levels correlating with reduced lesion sizes. However, cytokine therapies have not translated well to the clinic partially due their rapid clearance and pleiotropic nature. Methods: Here, we use protein engineering approaches to target IL-10 to LDL and subsequently to atherosclerotic plaques. We fuse IL-10 to an antibody fragment against oxidized LDL and show by surface plasmon resonance that it binds to both native LDL and oxidized LDL while retaining its ability to phosphorylate STAT3 in RAW 264.7 macrophage/monocyte-like cells. We then use flow cytometry to evaluate immune cell populations in the aortas of apolipoprotein E -/- mice on a high fat diet after 4 weekly injections. Results: Mice treated with our targeted IL-10 fusion protein had significantly reduced numbers of total leukocytes in the aorta compared to nontreated mice after 12 weeks on a high fat diet (n=6, Figure 1a). The number of macrophage/monocyte cells which may contribute to foam cell populations in the plaque was similarly reduced (Figure 1b). No differences in immune cell populations were observed in the spleen, indicative of the localized treatment effect. Furthermore, neither native IL-10 nor IL-10 fused to an antibody fragment of irrelevant specificity (non-targeted IL-10) showed any reduction of immune cell infiltration in the aorta. These findings were reproduced in apolipoprotein E -/- mice fed a high fat diet for 9 weeks (n=8) to show that aortic immune cell infiltration is also reduced at earlier stages in atherosclerosis progression. Conclusion: Engineered cytokines that specifically target LDL effectively provide local immunosuppression and show promise in the prevention and treatment of atherosclerosis.$

Targeted IL-10 locally suppresses inflammation in atherosclerotic sites

Abstract Atherosclerosis is a main contributor to cardiovascular disease, the leading cause of death in the U.S. Preventative measures include statins or dietary changes to reduce “bad cholesterol” or low-density lipoprotein (LDL) that accumulates on arterial walls to form plaques. However, these treatment options often lead to surgical interventions associated with high cost and morbidity. Although atherosclerosis is now recognized as a chronic inflammatory disease, no therapies targeting the underlying immunology are currently approved. The objective of this project is to locally suppress inflammation at atherosclerotic sites. IL-10 has been shown to suppress vascular inflammation, but its poor pharmacokinetic profile and possible side effects limit its therapeutic potential. To overcome these challenges, we engineered fusion proteins in which one side is IL-10 and the other side is an antibody fragment (Fab) that binds to protein epitopes of LDL. Thus, we specifically target IL-10 to atherosclerotic plaques. We validated the fusion proteins in vitro, confirming Fab binding affinity with surface plasmon resonance and IL-10 bioactivity in a macrophage-like cell line. We subsequently evaluated their immunosuppressive effect in the apolipoprotein E−/− mouse model of atherosclerosis. LDL-binding Fab-IL-10 constructs significantly reduced the total number of leukocytes, macrophages, T cells, and B cells in the aorta to levels similar to wild type mice. Conversely, neither IL-10 fused to a Fab of irrelevant specificity nor native IL-10 showed any differences to untreated mice. Therefore, plaque-targeted Fab-IL-10 constructs effectively suppress local inflammation and show promise as novel atherosclerosis therapies. Supported by NIH NHLBI T32, Chicago Immunoengineering Innovation Center

Depletion of γδ T Cells Leads to Reduced Angiogenesis and Increased Infiltration of Inflammatory M1-like Macrophages in Ischemic Muscle Tissue

γδ T cells, a small subset of T cells in blood, play a substantial role in influencing immunoregulatory and inflammatory processes. The functional impact of γδ T cells on angiogenesis in ischemic muscle tissue has never been reported and is the topic of the present work. Femoral artery ligation (FAL) was used to induce angiogenesis in the lower leg of γδ T cell depleted mice and wildtype and isotype antibody-treated control groups. Gastrocnemius muscle tissue was harvested 3 and 7 days after FAL and assessed using (immuno-)histological analyses. Hematoxylin and Eosin staining showed an increased area of tissue damage in γδ T cell depleted mice 7 days after FAL. Impaired angiogenesis was demonstrated by lower capillary to muscle fiber ratio and decreased number of proliferating endothelial cells (CD31+/BrdU+). γδ T cell depleted mice showed an increased number of total leukocytes (CD45+), neutrophils (MPO+) and neutrophil extracellular traps (NETs) (MPO+/CitH3+), without changes in the neutrophils to NETs ratio. Moreover, the depletion resulted in a higher macrophage count (DAPI/CD68+) caused by an increase in inflammatory M1-like macrophages (CD68+/MRC1−). Altogether, we show that depletion of γδ T cells leads to increased accumulation of leukocytes and M1-like macrophages, along with impaired angiogenesis.

Role of leucocytes in liver damage in experimental models of type 1 and 2 diabetes mellitus

Diabetes mellitus is a metabolic disorder, which results from insufficient secretion of insulin and/or its action, thus leading to hyperglycemia. Liver damage is known to be among the most common complications of type 2 diabetes mellitus (T2D) and is common in T1D. Comparison of the leukocyte phenotypes in liver tissue with appropriate blood parameters may assess degree of liver damage and search for approaches to correction of liver destruction in diabetes mellitus. Therefore, we aimed for assessment of changes in liver injury markers in blood and the numbers of leucocytes (CD45+cells), T lymphocytes (CD3+cells) and macrophages in the liver in experimental models of types 1 and 2diabetes. The experiment was conducted on 30 male Wistar rats. Alloxan at the dose of 170 mg/kg of body weight was used for T1D modeling. To provide a model of T2D, streptozotocin and nicotinamide were injected at the doses of 65 mg/kg, and 110 mg/kg respectively. Intact animals were used as a comparison control. Biochemical, hematological, immunohistochemical and morphometrical methods were used in the study. In T1D and T2D groups, levels of glucose (10.88±0.47 mmol/l and 10.78±0.42 mmol/l) and glycosylated hemoglobin (6.73±0.78% and 6.60±0.20% correspondingly) were rather close to each other and exceeded the values of intact rats (5.20±0.40 mmol/l and 4.07±0.30%). At the same time, the increase in total leucocyte number and fraction of peripheral blood leucocytes against normal levels were more pronounced in the T2D group than in T2D group. In liver of rats from the both diabetic groups, increased numbers of sinusoidal cells, macrophages, CD45+cells and CD3+ cells relative to intact rats were detected. However, in rats from T1D group, CD45+cells were distributed, mainly, in the liver parenchyma, whereas in rats in T2D group they showed sinusoidal location. At a similar degree of increasing macrophage numbers, and total CD45+cells number, higher counts of sinusoidal cells and CD3+cells, located both in the parenchyma and perivascular area, were found in rats of T2DM group compared with this parameter in T1DM group. An increase in ALT activity confirms a more significant damage to liver cells in animals of the T2DM group, whereas, in T1DM group, an increased AST activity and a less pronounced increase in ALT activity indicate uniformly distributed cytolysis. The results of our study showed, that, despite similar hyperglycemia level, the inflammatory process at the level of the whole organism and local inflammatory process in the liver are more pronounced in the T2DM group. A more significant severity of inflammatory process and liver damage corresponds to increase in sinusoidal cells and CD3+cell infiltration of liver tissue.

The effect of lipopolysaccharide extracted from Escherichia coli on total WBCs, granulocytes and on phagocytic activity in female rats

The present study was conducted to compare the total white blood cells count, granulocytes and phagocytic activity in female rats immunized with lipopolysaccharide (LPS) extracted from two different isolates of Escherichia coli, one normal isolate from the gut of healthy adult and the other was pathogenic strain isolated from patients suffering from urinary tract infection. The study was included 35 female albino rats divided randomly into seven groups, three groups injected with different concentrations of LPS of normal flora of E.coli 90, 120 and 150 microgram/kilogram body weight respectively and the other three groups were injected with the same concentrations of LPS extracted from pathogenic strain of E.coli, the last group (7th) represented the control group which given normal saline only. The results showed that the LPS from normal isolate causes a significant increase in the total number of leukocytes, granulocytes and phagocytic activity in animals immunized with the concentration of 120 µg/kg compared with the control group, and the concentration of 150 µg/kg of the same extract resulted in a high significant increase in phagocytic activity and the concentration of 90 µg/kg did not stimulate any significant differences in all the variables, while immunization with the extract of pathogenic strain increases the total leukocytes at a concentration of 90 µg/kg only with a significant increases in phagocytic activity at all concentration, and it also did not cause any significant differences in the granulocytes count.

The role of growth hormone and IFR-1 in the formation of leukocytic blood composition in the body of calves during cultivation

Purpose: to evaluate the biological properties of growth hormones (GH, IGF-1) in the dairy and vegetable periods of growing Holsteinized Black-and-White heifers, determined by their ability to regulate the age-related variability of the leukocyte composition of the blood.Materials and methods. The work was carried out in the conditions of the SPK «Koelginskoe» named after V. I. Shundeeva I. N. (Chelyabinsk region) in 2019–2020 on heifers of Holsteinized Black-and-White breed during the dairy and vegetable growing periods. Animals up to 3 months of age were kept individually, and then in groups of 10 animals. Milk and milk replacer were used in the diet of animals only up to 3 months, and then they were transferred to a plant-based diet. When compiling the feeding ration on the farm, they were guided by the norms of the VIZh. The experimental group (n=10) was formed according to the principle of approximate analogues, heifers were included in its composition after a clinical examination, which was carried out later with each blood sampling. For laboratory studies, whole blood and blood serum were used. Blood sampling was performed at 1, 3, 6, 9, 12 and 13 months of age in the morning before feeding. The leukocyte composition of the blood, the concentration of growth hormone and IGF-1 were determined.Results. It was found that the total number of leukocytes in the bloodstream of heifers increases with age by 28.06% (p&lt;0.05), which is accompanied by an increase in the percentage of eosinophils, segmented neutrophils and monocytes in the leukogram by 45.31; 5.70 and 50.00% (p&lt; 0.05), a decrease in lymphocytes, rodshaped neutrophils by 5.69 and 43.39% (p&lt;0.05). The level of STH by the end of the heifer rearing period decreases by 54.32% (p &lt;0.05), and the insulin-like growth factor type 1, on the contrary, increases by 32 times (p&lt;0.05). The values of the correlation coefficients in the statistical matrix of STG and IGF.

Higher Muscle Damage Triggered by Shorter Inter-Set Rest Periods in Volume-Equated Resistance Exercise.

ObjectivesThe aim of the manuscript was to analyze the effects of two rest periods between volume-equated resistance exercise (RE) on inflammatory responses (cytokines and leukocyte) and muscle damage.MethodsTen trained men (26.40 ± 4.73 years, 80.71 ± 8.95 kg, and 176.03 ± 6.11 cm) voluntarily participated in training sessions consisting of five sets of 10 reps performed at 10-RM on (1) the barbell bench press followed by (2) leg press, with either 1- or 3-min rest between sets and exercises. Circulating concentrations of different biomarkers was measured before (Pre), and after 3 h (excepted for cytokines), 6, 12, and 24 h from exercise. The rate of perceived exertion (RPE) was recorded after each set on both planned visits.ResultsWe found greater increases triggered by the 1-min rest period in Creatine Kinase (CK), occurring from 12 to 24 h post-exercise compared to the 3-min rest condition. A significant increase in the 1-min rest condition was also observed in the total number of leukocytes, neutrophils, and monocytes. The 1-min rest period also triggered increases compared to baseline in pro-inflammatory cytokines [Interleukin 1 beta (IL-1β), p = 0.004; tumor necrosis factor α (TNF-α), p = 0.01; and granulocyte-macrophage colony-stimulating factor (GM-CSF), p = 0.01], which were more evident after 6 and 12 h post-exercise. Similarly, increases in anti-inflammatory cytokines [Interleukin 5 (IL-5), p = 0.01; Interleukin 6 (IL-6), p = 0.01; and Interleukin 10 (IL-10), p = 0.01] at all time-points were observed.ConclusionOur results indicate that a 1-min rest condition in volume-equated RE promoted greater overall muscle tissue damage with a longer duration of the inflammatory processes compared to a 3-min rest.

Predictive Value of Immune Cells in the Risk of Gestational Diabetes Mellitus: A Pilot Study.

Immunological and biochemical parameters are gaining more and more importance in the prognosis of diabetes and its complications. Here, we assessed the predictive power of immune cells correlated with biochemical parameters in gestational diabetes mellitus (GDM). Immune cells and serum biochemical parameters were determined in women with GDM and pregnant controls. Receiver operating characteristics (ROC) curve analyses were conducted to assess the optimal cutoff and value of ratios of immune cells to biochemical parameters for predicting GDM. Blood glucose, total cholesterol, LDL-cholesterol and triglycerides were significantly increased whereas HDL-cholesterol decreased in women with GDM compared to pregnant controls. Glycated hemoglobin, creatinine, transaminase activities did not significantly differ between both groups. Total leukocyte, lymphocyte and platelet numbers were significantly high in women with GDM. Correlation tests showed that ratios of lymphocyte/HDL-C, monocyte/HDL-C and granulocyte/HDL-C were significantly higher in women with GDM than in pregnant controls (p = 0.001; p = 0.009 and p = 0.004 respectively). Women with a lymphocyte/HDL-C ratio greater than 3.66 had a 4-fold increased risk of developing GDM than those with lower ratios (odds ratio 4.00; 95% CI: 1.094 - 14.630; p=0.041). Our study showed that ratios of lymphocyte, monocyte and granulocyte to HDL-C might represent valuable biomarkers for GDM and in particular, lymphocyte/HDL-C ratio exhibited a strong predictive power for GDM risk.

The Impact of Anticoagulation Agent on the Composition and Phenotype of Blood Leukocytes in Dromedary Camels.

For the analysis of several cellular biomarkers, blood samples are anticoagulated using different agents with different modes of action. However, for the most commonly used anticoagulants, EDTA and heparin, varying effects on blood components have been reported in different species. As little is known about the impact of anticoagulants on the immunological evaluation of camel leukocytes, the present study analyzed the leukogram, the immunophenotype, and the cell vitality of camel leukocytes separated from blood samples anticoagulated with EDTA or lithium heparin. Using flow cytometry and staining with monoclonal antibodies to several cell surface markers, the composition and immunophenotype of camel leukocytes separated from blood anticoagulated with EDTA or heparin were analyzed. In comparison to EDTA-anticoagulated blood, using lithium heparin as an anticoagulant resulted in reduced numbers of total leukocytes and reduced numbers of neutrophils, which led to a reduced neutrophil to lymphocyte ratio. The analysis of cell necrosis and apoptosis after the staining of leukocytes with the DNA-sensitive dye propidium iodide and the mitochondrial membrane potential probe JC1 revealed a higher fraction of necrotic neutrophils and higher fractions of apoptotic neutrophils and monocytes in heparin blood than in EDTA blood. In addition, monocytes from heparin blood showed higher expression levels of the cell surface markers CD14, CD163, and MHCII when compared to cells from EDTA blood. Similarly, in heparin blood, CD44 and CD172a were expressed higher on neutrophils, while CD11a was expressed higher on lymphocytes in comparison to cells from EDTA blood. The results of the current study indicate the importance of considering the type of anticoagulant when investigating the composition, vitality, and immunophenotype of camel leukocytes.

Minthostachys verticillata Griseb (Epling.) (Lamiaceae) essential oil orally administered modulates gastrointestinal immunological and oxidative parameters in mice

Ethnopharmacological relevanceMinthostachys verticillata (Griseb.) Epling (Lamiaceae) is a plant used in folk medicine for digestive or respiratory disorders. In addition, it is incorporated as condiment, in foods, as beverage flavoring or mate. The ethnopharmacological interest of M. verticillata resides in its essential oil (EO). Part of group has demonstrated the immunomodulatory ability of EO giving this oil a biological potential not known until that moment and conducted studies to evaluate their possible application in diseases of veterinary interest. However, the immunomodulatory effects of EO administered orally have not been fully characterized. Aim of the studyThis study evaluated the impact of EO oral administration on gastrointestinal and immune health through measurement of immunological and oxidative parameters in mice. Material and methodsThe EO was extracted from the leaves, slender stems and flowers of M. verticillata by hydrodistillation and chemical analyzed by gas chromatography-mass spectrometry (GC-MS). Prior to in vivo study, the cytotoxic effect of EO was determined using the human colon carcinoma Caco-2 cell line. For in vivo study, three groups of male Balb/c mice (n = 3) were orally administered with saline solution (control group) and EO (5 or 10 mg/kg/day) during 10 consecutive days. Subsequently, histological and hematological parameters, cytokines production, oxidative markers and CD4+ and CD8+ T cells were evaluated. ResultsThe chemical analysis of EO revealed the presence of a high content of monoterpenes, being the main pulegone (76.12%) and menthone (14.28%). The EO oral administration improved mice growth performance and modulated systemic adaptive immune response by increasing in the total leukocyte number. A high percentage of CD4+ T cells were observed whereas the number of CD8+ T cells was not altered. EO did not alter the morpho-physiology of intestine and improved total antioxidant capacity by decreasing MDA concentrations. In addition, EO decreased the IL-6 levels and increased in the IL-4 and IL-10 concentrations. ConclusionResults indicate that M. verticillata EO modulate inflammatory and oxidative parameters constituting a natural alternative which could be applied to improve gastrointestinal and immune functionality in animals.

Серозные менингиты у детей: расширение диагностических возможностей

Обследованы 42 ребенка в возрасте 12 мес. — 10 лет, госпитализированных с предварительным диагнозом «менингит». Анализировали симптомы заболевания, результаты лабораторных исследований крови, ликвора, активность лактатдегидрогеназы (ЛДГ) ликвора. Тяжесть больных с серозными менингитами оценена по шкале тяжести AMSS. Была установлена статистически достоверно высшая активность лактатдегидрогеназы ликвора у пациентов с серозными менингитами. Активность ЛДГ зависела от уровня цитоза ликвора, концентрации в ликворе белка, общего количества лимфоцитов в периферической крови, что указывает на возможность использования исследования активности ЛДГ ликвора в качестве дополнительного показателя в диагностике серозных менингитов у детей.

Examination ofwhite blood cell indicators for three different ploidy level sturgeon species reared inanindoor recirculation aquaculture system for one year.

Functional diploid Acipenser ruthenus, functional tetraploid Acipenser gueldenstaedtii and functional hexaploid Acipenser brevirostrum juveniles were sampled monthly for one year, and the white blood cell indicators were determined. The total number ofleukocytes (TL) was 40.93± 17.24× 109/l for the diploids, 20.63± 11.20×109/l for the tetraploids, 14.13± 7.72×109/l for the hexaploids. The TL decreased with anincreasing ploidy level. The highest number ofleukocytes was reached during September and October for A.ruthenus and A.brevirostrum, from October toJanuary for A.gueldenstaedtii (astatistically significant finding). The lymphocytes dominated (76.89-80.14%) inthe differential counts and were found tobe reduced inJune and July ineach group. Granulocytes were represented byneutrophils and eosinophils. Counting from all the leukocytes, the neutrophils represented 13.0-18.7% and eosinophils represented 5.7-6.1%. Increasing number ofnuclear segments inthe granulocytes was dependent onthe increasing ploidy level. Nuclear segmentation inthe lymphocytes was acommon finding inhigher ploidy level groups. The data suggest asignificant effect ofploidy level onthe total number ofleukocytes and morphological nuclear changes inthe granulocytes and lymphocytes. The seasonal variation inthe differential leukocyte counts depends onthe species and the influence ofvarious external conditions rather than the ploidy level.

High C-Reactive Protein Levels Are Relatedto Better Survival in Patients with Uveal Melanoma.

PurposeTo determine whether peripheral blood leukocyte numbers and serum markers of inflammation can be used to predict which patients with primary uveal melanoma will develop metastasis.DesignRetrospective study.ParticipantsMedical records of patients with uveal melanoma (UM) who received treatment for primary UM between February 1992 and December 2020 at the Erasmus University Medical Center (Rotterdam, The Netherlands) and the Rotterdam Eye Hospital (Rotterdam, The Netherlands) were reviewed.MethodsInclusion criteria were the presence of a melanoma of the choroid or ciliary body and the availability of data from peripheral blood samples taken before treatment of the melanoma. Data including patient demographics, C-reactive protein (CRP) levels; erythrocyte sedimentation rate (ESR); number of leukocytes, neutrophils, monocytes, and lymphocytes; and histopathologic findings were obtained from medical records. Neutrophil-to-lymphocyte ratio (NLR) and lymphocyte-to-monocyte ratio (LMR) were calculated.Main Outcome MeasuresMetastasis-free survival.ResultsOf the 807 patients with UM, serum and leukocyte data were available for 183 of them at the time of primary tumor treatment. In the total group, no correlation was found between ESR before treatment; the number of leukocytes; percentages of neutrophils, monocytes, and lymphocytes; or NLR or LMR values and any of the clinical characteristics or metastasis-free survival. Among patients who underwent enucleation, those with negative BAP1 findings showed significantly lower numbers of leukocytes (P < 0.05). In the entire cohort, a significant association was found between high CRP levels and longer metastasis-free survival (MFS; P = 0.049).ConclusionsThe total blood leukocyte number was related to loss of BAP1 staining in patients who underwent enucleation, with lower leukocyte counts correlating with absent BAP1 staining. Higher CRP levels were associated with a longer MFS in the entire cohort. Neither the NLR nor the LMR is a good predictor for metastasis developing in patients with UM.

The anti-inflammatory mechanism of SAHA in acute pancreatitis through HDAC5/SLIT2/Akt/β-catenin axis.

Acute pancreatitis (AP) is widely recognized to be an inflammation-related disease, in which HDAC was upregulated. The anti-inflammatory role of suberoylanilide hydroxamic acid (SAHA), a HDAC inhibitor, has been documented. In this context, this research was implemented to figure out whether SAHA manipulated inflammation in AP. Subsequent to induction of AP mouse model, HDAC5 expression was detected. The binding of HDAC5 and SLIT2 was detected by Co-Immunoprecipitation and Chromatin immunoprecipitation assays. SAHA treatment and gain- and loss-of-function approaches were used in AP mice and lipopolysaccharide (LPS)-induced pancreatic acinar cells. In mice, biochemical methods were implemented to measure activities of pancreatic lipase, trypsin, myeloperoxidase (MPO) and pancreatic edema, TUNEL staining to determine pancreatic cell apoptosis, and flow cytometry to assess the total number of leukocytes and neutrophils in pancreas. In pancreatic acinar cells, CCK-8 was performed to evaluate cell viability. HDAC5 exhibited overexpression in AP mice. Mechanical analysis showed that HDAC5 facilitated SLIT2 deacetylation to downregulate SLIT2, thus activating Akt/β-catenin pathway in pancreatic acinar cells. SAHA treatment, HDAC5 silencing or SLIT2 overexpression diminished inflammation in AP in vivo and in vitro. SAHA treatment, HDAC5 silencing or SLIT2 overexpression reduced activities of pancreatic lipase, trypsin, MPO, pancreatic edema and cell apoptosis in AP mice as well as elevated viability of LPS-induced pancreatic acinar cells. SAHA might exert anti-inflammatory effects in AP mice via HDAC5/SLIT2/Akt/β-catenin axis.