Total Number Of Leukocytes Research Articles

Features of changes in nonspecific factors of immunological reactivity in obesity

Introduction. The study of pathogenetic factors of obesity is an urgent task of modern medicine. The formation of obesity is characterized by changes in the activity of individual mechanisms of innate immunity. At the same time, the values of laboratory indicators that characterize them are often within the current boundaries of the reference values of laboratory indicators of the immunity of a healthy person. This complicates the pathogenetic assessment of the mechanisms of nonspecific immunological reactivity in obesity and determines the need for further study of the characteristics of nonspecific immune defense factors in this pathology.Aim. To identify the features of changes in cellular and humoral factors of nonspecific immunological reactivity in obesity.Materials and methods. A single-center cross-sectional, one-time controlled study was conducted with the participation of 118 people, of which 87 people were obese patients (BMI 37.2 [34.1; 42.05] kg/m2), 31 people had normal body weight (BMI 21.9 [ 20.2; 23.5] kg/m2) and were included in the control group. All patients underwent a study of lipid profile (total cholesterol, high-density lipoproteins, low-density lipoproteins, very low-density lipoproteins, triglycerides), carbohydrate metabolism (glucose, insulin, glycated hemoglobin), C-reactive protein, indicators of cellular and humoral factors of nonspecific immunity (leukogram, cytokine profile, C3-C4 complement components).Results. An increase in the total number of leukocytes was revealed, due to neutrophil granulocytes against the background of the development of a disproportion between the percentage and absolute value of the number of lymphocytes and monocytes, the concentration of C3 and C4 complement components, C-reactive protein, as well as an increase in the level of IL-6, which confirms the presence of low-grade chronic inflammation in obese patients. Statistically significant correlations of immunological parameters with anthropometric data, indicators of carbohydrate and lipid metabolism were revealed.Conclusion. The results of the study indicate that obesity causes activation of certain cellular and humoral mechanisms of nonspecific immune defense involved in the formation of the inflammatory process. Confirmation of the presence of a latent inflammatory process in obesity is an increase in the level of leukocytes and their individual cellular forms, C-reactive protein, C3 and C4 complement components, IL-6. A feature of the changes is the presence of fluctuations in the values of the studied indicators within the current boundaries of the reference values of laboratory indicators, which makes it difficult to timely diagnose chronic inflammation in obesity

Wire injury induced moderate aortic valve stenosis in mice is accompanied by a chronic systemic inflammatory reaction

Abstract Background Patients with aortic valve stenosis (AS) have elevated serum levels of inflammation markers, and blood cytokine levels correlate with ventricular recovery after transcatheter aortic valve replacement. While presence of inflammatory processes in stenotic aortic valves is acknowledged, no systematic characterization of the systemic immune reaction upon AS has been performed, yet. Purpose Hypothesis of this study was that AS induces a systemic inflammatory reaction linked with local processes in the heart. Methods AS was induced by wire injury (WI) or sham surgery was performed in 3-4 months old male C57Bl/6J mice. AS severity, left ventricular (LV) function and hypertrophy indices were assessed with echocardiography (echo). After four weeks, explanted organs were weighted, and flow cytometry identified total leukocyte numbers in blood, heart, and peripheral immune organs (spleen, liver, lymph nodes) and uptake of Cy5-labelled perfluorocarbon nanoemulsions by whole blood leukocytes. Costimulatory molecules (flow cytometry, RT-qPCR) and cytokines (RT-qPCR, Multiplex Immunoassay) were analysed in heart, peripheral immune organs and blood. Results R-values were positive for spleen weight correlation with AS severity, cardiac function and mass suggesting cardiac hypertrophy to preserve heart function. Myocardial myeloid and T cell r-values were positive or negative, respectively, for correlation with spleen and liver weight, hinting at T cell recruitment from peripheral stores. Immune cell numbers in peripheral lymph nodes and spleen showed negative r-values for correlation with heart hypertrophy indices; number of liver myeloid cells correlated negatively with LV wall thickness (monocytes=-0.8571, p=0.0238) suggesting immune cell recruitment to hypertrophic hearts. Cytokine mRNA levels trended mainly towards increase in heart and regional lymph nodes and reduction in spleen and liver after WI. Correlation with echo was more homogeneous after WI, with r-values mainly positive, except of T cell activation markers, for aortic valves, but negative for myocardium, hinting at mechanisms preserving heart function. Correlating unchanged cytokine protein levels in myocardium and plasma with echo, r-values were mostly positive for myocardium, and were more positive than in sham for plasma. Echo and costimulatory molecule correlation showed a more homogeneous pattern in WI, with mostly positive r-values in myocardium and periphery, while most r-values were negative in sham. Together with reduced PFC-uptake by lymphatic cells, this hints at systemic immune cell activation in AS. Conclusion The results suggest that number and activation state of leukocytes in peripheral organs are directly linked to cardiac processes in AS. Considering the pathological value of inflammation, it is crucial to in future studies find out if modulation of the systemic inflammatory reaction relieves severity of AS and prevents development of heart failure.

Regulator of G-Protein Signaling 2 Knockout in CD4+ T Cells Promotes Anti-Inflammatory T Cells, Enhancing Ovulation, and Oocyte Yield.

To determine the downstream effects on ovarian function and immune cell differentiation in the ovary and uterus using a model in which RGS2 was knocked out specifically in CD4+ T cells. Laboratory based experiments with female mice. Female congenic (fully backcrossed) and non-congenic (mixed strain) mice with CD4 T cell-specific RGS2 knockout. Four-week-old female CD4 RGS2 knockout (CD4 RGS2 KO ) mice and their littermate controls (CD4 RGS2 CTL ) were subjected to superovulation using pregnant mare serum gonadotropins. Oocyte numbers, lymphocyte populations in the ovary and uterus, and serum estradiol and progesterone concentrations. In non-congenic (mixed strain) mice, CD4 RGS2 knockout (KO) promoted higher oocyte ovulation and increased uterine total leukocyte numbers. Similarly, congenic (fully backcrossed strain) mice showed higher oocyte numbers and increased uterine total leukocytes in the CD4 RGS2 KO mice compared to CD4 RGS2 CTL mice. Pro-inflammatory CD4+ T helper (T H ) 1 and T H 17 cell frequencies in the ovary and uterus were unchanged, while Treg and T H 2 cell frequencies were elevated, along with increased concentrations of estradiol and progesterone in the serum of CD4 RGS2 KO mice. Our study highlights the important role of RGS2 in CD4+ T cells within the context of reproduction. The dysregulation of immune responses due to RGS2 knockout in CD4+ T cells appears to enhance oocyte production. Further research is warranted to elucidate the precise mechanisms by which RGS2 influences reproductive outcomes, including its impact on fecundability, endometrial receptivity, and successful implantation.

Platelet Indices in Patients With Gram-Negative and Gram-Positive Sepsis: A Retrospective Cross-Sectional Study.

Different inflammatory processes and sepsis can significantly affect the number of platelets and platelet indices. Therefore, in this study, total platelet count (PLT), thrombocrit (Pct), platelet distribution width (PDW), mean platelet volume (MPV), and platelet-large cell ratio (P-LCR) were analyzed in patients with Gram-negative and Gram-positive bacterial sepsis and in sterile blood cultures. Inclusion criteria were an increased number of inflammatory parameters (elevated values ​​of leukocytes,C-reactive protein (CRP), procalcitonin (PCT), andpositive blood culture. Exclusion criteria were patients who did not have elevated values ​​of inflammatory parameters and did not have a positive blood culture.Samples were collected from patients who had sepsis confirmed by blood cultures at the Department of Microbiology and Molecular Diagnostics at University Clinical Hospital Mostar in the period from 2019 to 2022. Three groups were analyzed, patients who had sterile blood cultures, patients with blood cultures with isolated Gram-positive bacteria, and patients with blood cultures with isolated Gram-negative bacteria. Specific infectious agents were identified for each group of patients. In addition to the above, PLT, Pct, MPV, PDW, P-LCR, PCT, CRP, the total number of leukocytes, and the number of neutrophil leukocytes were analyzed in each group. The values of PCT, CRP, and the number of neutrophile leukocytes were significantly higher in patients with Gram-negative sepsis as compared to Gram-positive sepsis and to control group. Patients with sepsis have decreased PLT and Ptc and increased values of MPV, PDW, and P-LCR. In sepsis caused by the Gram-negative bacteria, i.e., Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii, the values of the same parameters were more changed compared to sepsis caused by Gram-positive bacteria, i.e., Streptococcus pneumoniae, Enterococcus spp., and methicillin-resistant Staphylococcus aureus (MRSA). When comparing Gram-negative negative bacteria, PLT was lowest in sepsis caused by Escherichia coli, the PDW value was highest in sepsis caused by Acinetobacter baumannii, and MPV and P-LCR were the highest in sepsis caused by Klebsiella pneumoniae. Our study showed that platelet indices are significantly changed in patients with sepsis. Patients with sepsis have decreased values of PLT and Pct and increased values of MPV, PDW, and P-LCR, indicating an increase in thrombocyte production. Moreover, the results were more prominent in sepsis caused by Gram-negative bacteria compared to sepsis caused by Gram-positive bacteria.

Cyperus articulatus: Anti-inflammatory and antinociceptive activity of a medicinal plant from the Amazon

Ethnopharmacological relevanceCyperus articulatus L., popularly known as priprioca, is a plant used in the Amazon for perfumed baths and homemade perfumes. In traditional medicine, its rhizomes are used to treat diseases related to inflammatory processes. Aim of the studyDue to its promising bioactive properties, this study sought to investigate its phytochemistry and the anti-inflammatory and antinociceptive activity of the essential oil obtained from C. articulatus (CAEO) in in vitro and in vivo tests. Material and methodsThe essential oil was obtained from the rhizomes of C. articulatus and extraction was carried out via hydrodistillation. Then, the oil was analyzed by GC-MS analyses. Initially, culture of RAW 264.7 macrophages stimulated by lipopolysaccharide (LPS) was used to evaluate cytotoxicity and interference in the production of mediators of the inflammatory process (nitrite, IL-1β, TNF-α and PGE2) after exposure to CAEO. The acute toxicity of CAEO was evaluated and the results were used to define doses of 10, 100 and 400 mg kg−1 for evaluation of CAEO in in vivo tests using mice. The carrageenan-induced air pouch models and the Evans test were used to evaluate the anti-inflammatory activity by measuring the number of total leukocytes and vascular permeability. Antinociceptive activity was evaluated via tests of contortions induced by acetic acid, hot plate, and formalin. ResultsTreatment with CAEO reduced the levels of nitrite IL-1β, TNF-α and PGE2 in the macrophage culture, revealing its anti-inflammatory potential. CAEO decreased carrageenan-induced leukocyte migration and vascular permeability, which are important events related to the acute inflammatory response. Nociceptive activity was significantly inhibited by CAEO in the acetic acid-induced contortions model, hot plate, and in both phases of the formalin test. The treatment with naloxane reversed the antinociceptive effect observed in the formalin test, suggesting the participation of opioid receptors in the mechanism of action of CAEO. ConclusionThe observed results reveal the anti-inflammatory and antinocipeptive activity of C. articulatus essential oil in vivo and support the traditional use of this plant in the treatment of different diseases involving inflammation and pain.

Tenacibaculum maritimum can boost inflammation in Dicentrarchus labrax upon peritoneal injection but cannot trigger tenacibaculosis disease.

Despite being a bacterial pathogen with devastating consequences, Tenacibaculum maritimum's pathogenesis is not fully understood. The aim of the present study was to elucidate if different inoculation routes (intraperitoneal - i.p - injection and bath challenge - known to induce mortality) can induce tenacibaculosis (i.e., using the same T. maritimum inoculum), as well as evaluate the short-term immune response of European sea bass (D. labrax). Additionally, the host response against i.p. injection of extracellular products (ECPs) was also studied. Fish were i.p. challenged with 5.5 × 105 CFU mL-1 of T. maritimum cells with or without ECPs (BECPs and BWO, respectively), ECPs alone or marine broth (mock). Another group of fish was bath-challenged with 5.5 × 105 CFU mL-1 to confirm the virulence of the bacterial inoculum. Undisturbed specimens were used as controls. The severity of both challenges was determined by following percentage survival. Blood, liver and head-kidney samples were collected at 0, 3, 6, 24 and 48h post-challenge for assessing immune parameters, oxidative stress and gene expression. Total and differential peritoneal cell counts were performed. The presence of viable bacteria in the blood and peritoneal cavity was studied. Symptoms of tenacibaculosis, such as skin/fin abrasions, were only observed in the bath-challenged fish, where 0% survival was recorded, whereas 100% survival was observed after i.p. injection of the same bacterial inoculum. An increase in total leukocyte numbers in the peritoneal cavity was observed 3h post-injection of BECPs when compared to the other treatments. Blood total leukocytes, lymphocytes, and thrombocyte numbers dropped after the challenge, mainly in fish challenged with BECPs. At 48h post-challenge, bactericidal activity in the plasma increased in fish injected with bacteria (with and without ECPs). The same tendency was seen for some of the oxidative stress parameters. The increased expression of il1β, il6, il8, and hamp1 in fish challenged with ECPs and BECPs suggests a more exacerbated pro-inflammatory response in the head-kidney against these inocula. The infection trial and the observed immune responses showed that the infection route is a determinant factor regarding T. maritimum-induced pathogenesis in European sea bass.

UHPLC-DAD/ESI-TOF-MS Phytochemical Characterization and Evaluation of the Impact of Eleutherococcus senticosus Fruit Intractum on Biochemical, Hepatological, and Blood Parameters in Balb/c Mice.

Eleutherococcus senticosus (Rupr. et Maxim.) Maxim. (ES) has gained popularity for its adaptogenic, immunostimulant, and anti-inflammatory properties. Because of overexploitation of the roots, the species is considered to be endangered and has been put on the Red List in some countries (e.g., the Republic of Korea). Therefore, the fruits of E. senticosus might be explored as a new sustainable source of compounds with adaptogenic activity. This study aimed to assess the chemical composition and the safety profile (hepatotoxicity, blood morphology, biochemical parameters of blood plasma) of E. senticosus fruit intractum in Balb/c mice after oral administration of 750 and 1500 mg/kg b.w. UHPLC analysis coupled with DAD and MS detectors was used to quantify the metabolites. For the first time, oleanolic and ursolic acids were quantified in the intractum (16.01 ± 1.3 and 2.21 ± 0.17 µg/g of oleanolic and ursolic acids, respectively). Regarding polyphenols, chlorogenic acid (0.92 mg/g of dried extract), caffeic acid (0.43 mg/g), dicaffeoylquinic acids (in total: 1.27 mg/g), and an unidentified caffeic acid ester (0.81 mg/g) were identified. The results in Balb/c mice revealed that the intractum does not cause significant variations in red blood cells parameters. In turn, a significant decrease in the total number of leukocytes was observed (5.8 × 103 µL), with a percentage increase in lymphocytes among the groups (80.2, 81.8, and 82.6). The ability of the intractum to decrease alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels may indicate its anti-inflammatory activity. Our observations justify that the fruits of E. senticosus are safe in the doses used and do not cause significant changes in the activity of the liver enzymes or in blood parameters.

Phytochemical Profile, toxicity assessment, and biological activities of hydroalcoholic extract of Calycophyllum spruceanum leaves

Pharmacotherapy for the treatment of inflammation based on non-steroidal anti-inflammatory drugs (NSAIDs) and opioids can cause gastrointestinal discomfort, ulcers, renal, cardiac and metabolic dysfunctions. Studies on natural products containing constituents with anti-inflammatory and analgesic activity remain important and quite justified. Plants belonging to the Rubiaceae family are examples of source of bioactives. The hydroalcoholic extract of Calycophyllum spruceanum (Benth.) Hook. F. ex K. Schum (Mulateiro) leaves (HECsL), native to the Amazon biomes, when subjective to liquid chromatography in UHPLC-MS, revealed the presence of four phenolic constituents: caffeoylquinic derivatives, small organic acids and flavonoids, constituents that have antioxidant and anti-inflammatory activity. HECsL did not show acute toxicity, nor did it change the motor coordination or body mass of the experimental animals. HECsL at a dose of 60 mg/kg reduced the number of writhings induced by 0.8% acetic acid (63%), increased the time spent on the hot plate (30%), and increased the tail withdrawal latency at a dose of 120 mg/kg (49%). The stimulation of hypernociception by carrageenan in the animals abdomen was reversed by HECsL (59%) by decreasing the number of total leukocytes (67%). HECsL has anti-inflammatory and antinociceptive prophylactic effects on the peripheral and central nervous system of Swiss mice, being devoid of acute toxicity.

Features of reactions to stress in rats with passive-defense behavior after light desynchronosis and physical activity

An assessment was made of hematological parameters of peripheral blood and structural and functional changes in the adrenal glands in stress-unresistant rats (passive-defensive type of behavior in the open field test) after exposure to light deprivation and physical activity during the spring equinox. The experiment showed that light deprivation for 10 days reduces the total number of leukocytes, the absolute content of monocytes, granulocytes, lymphocytes and the level of corticosterone in the peripheral blood compared to the intact group, that was on a natural lighting regime. Histological analysis of the adrenal glands of this group showed, that in the zona fasciculata under such conditions there was nuclear hypertrophy, an increase in the nuclear-cytoplasmic ratio of adrenocorticocytes and a decrease in the size of the zona glomerulosa of the adrenal cortex of rats in relation to the intact group. Physical activity in the form of forced swimming until complete fatigue for 5 days in a row in natural light in stress-unresistant rats did not change the parameters of the peripheral blood of animals, however it led to an increase in the area of the cytoplasm, the nuclear-cytoplasmic ratio, as well as the formation of hypertrophy of the nuclei of adrenocorticocytes in the fascicle adrenal zones, which indicated the preparation of cells for increased synthetic activity. Keeping rats for 10 days in complete darkness before forced swimming every day for 5 days, on the one hand, formed a hypoxic state and exhaustion of the adrenal glands, on the other hand, stabilized the leukocyte pool of peripheral blood compared to similar indicators in intact rats.

Differential composition and yield of leukocytes isolated from various blood component leukoreduction filters

In Flanders, an estimated 300,000 leukoreduction filters are discarded as biological waste in the blood establishment each year. These filters are a possible source of fresh donor leukocytes for downstream purposes including research. We investigated leukocyte isolation from two types of filters either used for the preparation of platelet concentrates (PC-LRF) or erythrocyte concentrates (EC-LRF). Outcome parameters were leukocyte yield, differential count, turnaround time and effect of storage conditions. Leukocytes were harvested by reverse flow of a buffer solution. Control was the gold standard density gradient centrifugation of buffy coats. Total leukocyte number isolated from PC-LRF (1049 (± 40) x 106) was almost double that of control (632 (± 66) x 106) but the differential count was comparable. Total leukocyte number isolated from EC-LRF (78 (± 9) x 106) was significantly lower than control, but the sample was specifically enriched in granulocytes (81 ± 4%) compared to control (30 ± 1%). Isolation of leukocytes from either PC- or EC-LRF takes 20 min compared to 240 min for control density gradient centrifugation. Leukocyte viability is optimal when harvested on day 1 post donation (95 ± 0.9%) compared to day 3 (76.4 ± 2.4%). In conclusion, our study demonstrates that leukoreduction filters from specific blood component processing are easy to use and present a valuable source for viable leukocytes of all types.

Anti-tumoral Immunity and Chemo-preventive Effectiveness of Herbal Extracts of Curcumin, Ginger, Clove and Amygdaline in Ehrlich Ascites Carcinoma-Challenging Mice.

Due to its systemic toxicity, traditional chemotherapy of tumors is being taken into consideration. Herbal therapy, containing phytochemical polyphenol derivatives such as Curcumin (Cur), Ginger (Gin), Cloves (Clov) and Amygdaline (Amyg), is one of the numerous complementary and alternative approaches as an anti-cancer therapy and holds great promise for cancer chemo-prevention with fewer side effects. The current study was designated to assess anti-tumoral immunity and anti-cancer and chemo-preventive effectiveness of herbal extracts of Cur, Ginger, Clov and Amyg in Ehrlich Ascites Carcinoma (EAC)-challenging mice. Chemo-preventive efficacy of herbal extracts of Cur, Gin, Clov and Amyg were analyzed in vivo by examination of the apoptosis rate of EAC tumor cells by flow cytometry. The total numbers of EAC cells, splenocytes counts and leucocytes count with their differentials relative % in peripheral blood (PB) of EACchallenging mice were investigated. EAC-challenging mice treated with herbal extracts of Cur, Gin, Clov and Amyg showed a marked decline in EAC tumor cell count and a noticeable increase in apoptosis rate of EAC tumor cells, a remarkable decrease in serum level of cancer antigen 125 (CA-125) with an obvious increase in the number of splenocytes comparing to that in EAC-challenging mice treated with PBS alone. Moreover, the data indicated an insignificant change in the total leucocytes count and their differentials relative % of eosinophil, neutrophils, monocytes and lymphocytes in EAC-challenging mice treated with Cur and Amyg, but these parameters were markedly increased in EAC-challenging mice injected with Gin and Clov compared to that in EAC-challenging mice treated with PBS alone. To conclude, the herbal extracts of Cur, Gin, Clov and Amyg may have anti-tumoral immunity and anti-cancer potency and potential to reduce the resistance to cancer conventional chemotherapy and exert cancer chemo-protective approaches with low adverse effects. Further research is necessary to determine the regimen's toxicity on various tissues and organs and to connect the diagnostic and therapeutic approaches used in the regimen's biomedical use.

Immunological disorders in patients with acute pleural empyema

In case of acute pleural empyema pronounced violations of cellular and humoral factors of immunity as well as non-specific resistance of the body are revealed. Dynamics of laboratory and immunological changes of 64 patients with acute pleural empyema who were treated with classical and minimally invasive surgical methods has been studied. Patients were classified into 5 groups in accordance with severity of the disease, which was determined according to the criteria which comprise such clinical and laboratory parameters as: respiratory rate, heart rate, blood pressure, temperature reaction, the number of affected particles, leukocytosis, SpO2. All the patients underwent a standard examination as well as determination of serum concentration InterLeykin 6 (IL-6), InterLeykin 8 (IL-8), Tumor Necrose Factor α (TNF-α), C-Reactive Protein (CRP). Assessment of cytokine spectrum of blood serum of patients with acute pleural empyema allows stating the condition of hypercytokinemia with an increase in pro-inflammatory cytokines. Meanwhile the degree of their increase differed depending on the severity of course of acute pleural empyema. Study of structure of cytokine status allowed identifying those patients with severe course of pleural empyema had a significant increase in IL-6 and CRP concentrations. Nevertheless, the severe course of the disease is associated with insufficient increase of IL-8 and TNF-α. Deficiency of IL-8 and to a less extent IL-6 is expressed in patients of the older age group. In the general blood test, there have been identified higher indicators of total number of leukocytes, neutrophils, pronounced shift of the leukocyte formula to the left, increases of erythrocyte sedimentation rate in patients with a severe course of the disease. There have been identified factors which affect severe course of acute pleural empyema: decrease in oxygen saturation to less than 94%, severity of shortness of breath, which exceeds 2 points on the Borg scale, damage to 3 or more segments of lung tissue, and also numerous clinical signs of violation of anti-infective protection, pronounced decrease in neutrophils, an increase in the concentration of C-reactive protein and insufficient increase of IL-8 and TNF-α in blood serum. Keywords: acute purulent diseases of the pleura, respiratory failure, interleukin 6, interleukin 8, tumor necrosis factor α, C-reactive protein.

Diagnostic studies for enterotoxaemia in rabbits

The relevance of this study is highlighted by the occurrence of enterotoxaemia in rabbits, particularly caused by Clostridium perfringens, in the context of industrial rabbit farming. The research aimed to determine the causes of morbidity and increased mortality in young rabbits during the growing period. The study involved a stepwise analysis of feed samples, clinical examination of rabbits reared in an industrial rabbit farm, and laboratory examination of the obtained biological material. Additionally, to establish a diagnosis and provide the farm with appropriate recommendations regarding the maintenance and prevention of rabbit diseases, the health status of the rabbits, the causes of digestive disorders with symptoms of diarrhoea and increased mortality under production conditions were analysed. Clinical, haematological, pathological, microbiological, and statistical research methods were used. In diseased rabbits, an increase in rectal body temperature, symptoms of diarrhoea, and in some animals, seizures were established. Several animals succumbed to the disease. During laboratory blood tests of the rabbits, moderate anaemia was detected, with a decrease in the number of erythrocytes and a reduction in haemoglobin content. There was also a disturbance in the qualitative composition of erythrocytes, including the presence of poikilocytosis and altered erythrocyte forms: acanthocytes, echinocytes, schistocytes, dacryocytes, keratocytes, and drepanocytes. A decrease in the immune status of the rabbits was characterised by a reduction in the total number of leukocytes and lymphocytes in the blood and a low neutrophil-to-lymphocyte ratio. Biochemical analysis of rabbit serum revealed a decrease in glucose levels below the physiological range and an increase in the activity of the enzymes alanine aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase. Results of post-mortem examinations of the deceased rabbits indicated the presence of gas-filled small intestines, with a fluid content and no macroscopic signs of inflammation. The feed provided to the rabbits on the farm was found to contain sulphite-reducing clostridia, specifically Clostridium perfringens. Identifying the causes of morbidity and mortality in rabbits will enable the selection of appropriate methods for preventing deaths within specific farming conditions

The effect of exercise and disease status on mobilization of anti-tumorigenic and pro-tumorigenic immune cells in women with breast cancer.

Mobilization of certain immune cells may improve the ability of the immune system to combat tumor cells, but the effect of acute exercise on mobilizing immune cells has been sparsely investigated in cancer patients. Therefore, we examined how acute exercise influences circulating immune cells in breast cancer patients. Nineteen newly diagnosed breast cancer patients aged 36-68 performed 30 minutes of moderate-intensity exercise with a cycle ergometer. Blood samples were collected at various time points: at rest, at 15 (E15) and 30 minutes (E30) after onset of the exercise, and at 30 and 60 minutes post-exercise. We analyzed several immune cell subsets using flow cytometry. Acute exercise increased the number of total leukocytes, neutrophils, lymphocytes, monocytes, basophils, total T-cells, CD4+ T-cells, T helper (Th) 2-cells, Th 17-cells, CD8+ T-cells, CD4-CD8- T-cells, CD56+ natural killer (NK) cells, and CD14-CD16+ monocytes. Many of the changes were transient. Proportions of NK-cells and CD8+ T-cells increased, while the proportion of myeloid derived suppressor cells (MDSCs) reduced, and proportion of regulatory T-cells remained unchanged by exercise. Several associations were detected between cell mobilizations and disease state. For instance, tumor size correlated negatively with NK cell mobilization at E15, and progesterone receptor positivity correlated negatively with CD8+ T-cell mobilization. The findings show that the proportions of CD8+ T-cells and NK cells increased and the proportion of MDSCs proportion decreased in breast cancer patients after 30-minute exercise, suggesting a change in the profile of circulating immune cells towards more cytotoxic/anti-tumorigenic. The mobilization of some immune cells also appears to be related to the disease state.

A mixture of free and microencapsulated essential oils combined with turmeric and tannin in the diet of cattle in the growing and finishing phase: A new tool to enhance productivity

The study was conducted to evaluate whether adding a blend containing free and microencapsulated essential oils, combined with turmeric and tannin, can replace monensin as a performance improver, positively influencing animal health and meat quality. Fourteen male cattle with an average initial body weight of 253 kg and ten females with an average initial body weight of 239 kg were included in the experiment in a confinement system. The animals were divided into four independent groups: Males (M-PHYTO, inclusion of the phytogenic mixture; M-MONEN, inclusion of monensin); Females (F-PHYTO, inclusion of phytogenic; F-MONEN, inclusion of monensin). The diets were formulated for an average daily gain of 1.5 kg, with food provided twice daily. Data and biological samples were collected on days 33, 100, and 153. M-PHYTO and F-PHYTO showed greater average daily gain and body weight, with a tendency to consume more dry matter and feed efficiency. However, the apparent digestibility of nutrients was higher in animals in the MONEN group than those in the PHYTO group, regardless of sex. The total number of leukocytes in cattle in the M-PHYTO and F-PHYTO groups was higher due to the higher lymphocyte count in the blood of these animals. The activity of gamma-glutamyltransferase and aspartate aminotransferase in animals that consumed the phytogenic mixture was lower compared to animals that consumed monensin. TBARS levels were lower in the serum and meat of cattle in the M-PHYTO and F-PHYTO groups, as were the total thiol concentration and glutathione S-transferase activity in the blood and meat of these animals. Bacterial activity was greater in the rumen fluid of cattle that consumed the phytogenic, as demonstrated by the methylene blue reduction test. The total production of short-chain fatty acids and acetic acid showed an interaction between treatment and days for both sex classes and an effect of treatment for females, with a higher concentration in the rumen in the M-PHYTO and F-PHYTO groups. Propionic acid affected treatment and the treatment x day interaction for males and females, with the M-PHYTO and F-PHYTO groups showing higher concentrations. There was an interaction between treatment and day in the acetate/propionate ratio, being lower in M-PHYTO and F-PHYTO cattle. Treatment between ruminal microbiota groups had no effect. The rib eye area had more significant gains when consumed phytogenic by males and females. The subcutaneous and sirloin fat thickness was greater in F-PHYTO animals than in the control group; however, in males, the result was the opposite. The sum of polyunsaturated fatty acids in meat was higher in M-PHYTO compared to M-MONEN and lower in F-PHYTO meat compared to F-MONEN. These results indicate that the phytogenic mixture is a potential performance improver in the diet of growing and finishing cattle, regardless of sex, but meat characteristics differ between males and females.

The impact of grape pomace extract rich in natural complex of polyphenols on morpho-functional state of leukocytes under experimental diabetes mellitus

Background. Diabetes mellitus is one of the most common diseases in the world. Under this pathology all organs and systems of an organism are damaged, inclu­ding the immune system. Peripheral blood leukocytes are an important element of this system that suffer damage under diabetes mellitus due to the influence of reactive oxygen species and reactive nitrogen species, the number of which increases fast and leads to the development of oxidative-nitrative stress. Thus, the discovery of new diabetes-correcting drugs that possess hypoglycemic, antioxidant and immunomodulatory proper­ties is one of the principal tasks. Such properties are inherent in polyphenolic compounds, a large amount of which is contained in the grape pomace. That is why the study of grape pomace extract, rich in a natural complex of polyphenols, is important to evaluate the possibility of further use of these substances as a basis for drugs that can be used in the complex therapy of diabetes mellitus. Materials and Methods. The research used peripheral blood leukocytes of the control rats, the control animals that were treated with grape pomace extract for 14 days, animals with streptozotocin-induced diabetes mellitus and rats with experimental diabetes mellitus that were treated with grape pomace extract for 14 days. To evaluate the corrective effect of the grape pomace extract rich in a natural complex of polyphenols on the state of the antioxidant defense and the L-arginine/NO systems, the activity of antioxidant enzymes, the level of oxidative modification products of proteins and lipids, the activity of NO-synthase, the content of nitrites and nitrates, 3ʹ-nitrotyrosine-modified proteins were studied. The total number of leukocytes, the white blood cell differential and the phagocytic activity, and the average cytochemical coefficients of cationic proteins and NADPH-oxidase activity were indicated to study the effect of the grape pomace extract on the functional state of leukocytes. Results and Discussion. The study revealed normalization of the total number of leukocytes and the white blood cell differential, the activity of NADPH-oxidase, superoxide dismutase, catalase and glutathione peroxidase, and the levels of the oxidative modification products of proteins and lipids, nitrites, nitrates and 3ʹ-nitrotyrosine-modified proteins, as well as an increase in the average cytochemical coefficient of cationic proteins, and a decrease of the inducible NO-synthase activity after grape pomace extract administration for 14 days to animals with streptozotocin-induced diabetes mellitus. Conclusion. The obtained results confirm the antioxidant and immunomodulatory effects of the studied extract and justify the feasibility of using grape pomace complex of polyphenolic compounds as a basis for new drugs that will be used in the complex therapy of diabetes mellitus.

Analysis of Immune Cell Subsets in Peripheral Blood from Patients with Engineered Stone Silica-Induced Lung Inflammation.

Silicosis caused by engineered stone (ES-silicosis) is an emerging worldwide issue characterized by inflammation and fibrosis in the lungs. To our knowledge, only a few reports have investigated leukocyte/lymphocyte subsets in ES-silicosis patients. The present study was designed to explore the proportions of the main lymphocyte subsets in ES-silicosis patients stratified into two groups, one with simple silicosis (SS) and the other with a more advanced state of the disease, defined as progressive massive fibrosis (PMF). The proportions of B (memory and plasmablasts) cells, T (helper, cytotoxic, regulatory) cells, and natural killer (NK) (regulatory and cytotoxic) cells were investigated by multiparameter flow cytometry in 91 ES-silicosis patients (53 SS patients and 38 PMF patients) and 22 healthy controls (HC). Although the total number of leukocytes did not differ between the groups studied, lymphopenia was observed in patients compared to healthy controls. Compared with those in healthy controls, the proportions of memory B cells, naïve helper T cells, and the CD4+/CD8+ T cells' ratio in the peripheral blood of patients with silicosis were significantly decreased, while the percentages of plasma cells, memory helper T cells, and regulatory T cells were significantly increased. For the NK cell subsets, no significant differences were found between the groups studied. These results revealed altered cellular immune processes in the peripheral blood of patients with ES-silicosis and provided further insight into silicosis pathogenesis.

Hematological and genotoxic biomarkers in a natural population of freshwater turtles Trachemys dorbigni (Duméril & Bibron, 1835) (Testudines: Emydidae) living in an urban area in Southern Brazil.

The present study aimed to report the morphometric and hematological indices and genotoxicity of a free-life population of D'Orbigny's slider turtles (Trachemys dorbigni) living in an urban area in Southern Brazil. For that, 16 specimens were randomly captured in an urban canal that receives irregular releases of wastewater. Biometrics and external visual changes were analyzed, such as turtle shell deformities, and the presence of parasites. Blood samples were collected to evaluate the hematological profile and the presence of micronuclei and other erythrocyte nuclear abnormalities as potential mutagenic and genotoxic effects. Water physicochemical parameters were also measured. Organisms with ectoparasites (31.25%) and small carapace deformations (56.25%) were observed, but maximum carapace length and weight were considered normal for the species according to the literature. The blood profile indicated low hemoglobin and hematocrit and a high number of total leukocytes, particularly eosinophils which characterize parasitic infections. A frequency of 0.12% for the micronucleus was considered basal, but the frequency of other erythrocyte abnormalities was evident, mainly of blebbed nuclei (63.79%), indicating chromosomal damage in the early stage. The results of this study suggest that natural populations of chelonian inhabiting urbanized areas are impacted by anthropogenic activities in the surrounding environment. Furthermore, it provides comprehensive data which can serve as a comparative model for environmental monitoring studies involving turtles.

Prevalence of human herpesvirus in plasma and saliva of cirrhotic patients: A pilot study.

The objective of this study was to identify the presence of human herpesvirus (HHV) in the plasma and saliva of hepatic-cirrhosis patients and correlate it with clinical data and laboratory tests. This is a pilot, observational, and cross-sectional study. Specimens of plasma and saliva from 72 cirrhotic individuals were analyzed by means of polymerase chain reaction. The patient population had a mean age of 54.84 years old (SD±10) and was 70% males (51/72). Approximately 47% (n=34) of the patients had leukopenia and HHV was not identified in the plasma specimens. The main species of HHV identified in the saliva were HHV-7 (n=42, 62%) and Epstein-Barr virus (EBV) (n=30, 41%). Moreover, there was a significant decrease in the total number of leukocytes and lymphocytes in saliva containing EBV (P=.038 and P=.047, respectively). The results show that the presence of EBV in the saliva of cirrhotic patients was correlated with their circulating immune status. It may be possible that the immune dysfunction displayed by the cirrhotic patients plays a role in the shedding of EBV into saliva.

English

Excessive intake of a high-fat diet (HFD) is profoundly linked with various diseases, including dyslipidemia, inflammation, obesity, type 2 diabetes mellitus (T2DM), and inflammation. Meanwhile, Mentawai taro, a variety of taro in the Colocasia esculenta species, has traditionally served as a daily staple food for the local people in the Mentawai Islands (West Sumatra, Indonesia). However, it remains unclear whether Mentawai taro corm can effectively counteract the detrimental effects of an HFD. This study investigated whether Mentawai taro corm flour (MTF) can significantly prevent HFD-induced T2DM and leukocytosis. The experimental study involved adult male mice (n = 27) divided into three groups: normal diet, HFD, and HFD supplemented with 20% MTF. After a 12-week treatment, the levels of blood glucose, plasma insulin, glucose tolerance, insulin tolerance, homeostasis model assessment for insulin resistance (HOMA-IR), malondialdehyde (MDA) levels as well as leukocyte counts were determined. Results showed that supplementation of 20% MTF in the HFD substantially improved blood glucose profiles, reduced blood glucose levels (P < 0.01), enhanced glucose and insulin tolerance (P < 0.01) and prevented hyperinsulinemia and insulin resistance. Additionally, Mentawai taro flour effectively reduced pancreatic tissue MDA (P < 0.05) and regulated leukocyte profiles, including total leukocyte number (P < 0.01), and numbers of lymphocytes, monocytes, and granulocytes (P < 0.01). Hence, our study suggests that MTF supplementation in the context of an HFD exerts beneficial effects on glucose homeostasis, insulin sensitivity, oxidative stress, and inflammation. It implies potential antidiabetic and anti-inflammatory properties of MTF to manage diet-induced diseases