Blood Thrombocytes Research Articles

Association of Biological Age with Tumor Microenvironment in Patients with Esophageal Adenocarcinoma

Introduction: Esophageal cancer is the seventh most common cancer worldwide and typically tends to manifest at an older age. Marked heterogeneity in time-dependent functional decline in older adults results in varying grades of clinically manifest patient fitness or frailty. The biological age-related adaptations that accompany functional decline have been shown to modulate the non-malignant cells comprising the tumor microenvironment (TME). In the current work, we studied the association between biological age and TME characteristics in patients with esophageal adenocarcinoma. Methods: We comparatively assessed intratumoral histologic stroma quantity, tumor immune cell infiltrate, and blood leukocyte and thrombocyte count in 72 patients stratified over 3 strata of biological age (younger <70 years, fit older ≥70 years, and frail older adults ≥70 years), as defined by a geriatric assessment. Results: Frailty in older adults was predictive of decreased intratumoral stroma quantity (B = −14.66% stroma, p = 0.022) relative to tumors in chronological-age-matched fit older adults. Moreover, in comparison to younger adults, frail older adults (p = 0.032), but not fit older adults (p = 0.302), demonstrated a lower blood thrombocyte count at the time of diagnosis. Lastly, we found an increased proportion of tumors with a histologic desert TME histotype, comprising low stroma quantity and low immune cell infiltration, in frail older adults. Conclusion: Our results illustrate the stromal-reprogramming effects of biological age and provide a biological underpinning for the clinical relevance of assessing frailty in patients with esophageal adenocarcinoma, further justifying the need for standardized geriatric assessment in geriatric cancer patients.

IFN1 Enhances Thrombocyte Phagocytosis through IFN Receptor Complex-JAK/STAT-Complement C3.3-CR1 Pathway and Facilitates Antibacterial Immune Regulation in Teleost.

Type I IFNs with strong positive charges exhibit robust bactericidal activity and a protective effect against bacterial infections. However, the antibacterial mechanism invivo remains unknown. In this study, Ab blockade of IFN1, a member of type I IFNs in grass carp (Ctenopharyngodon idella), resulted in high mortality, tissue bacterial loads, and low expression of immune factors after bacterial challenge, which indicates that the antibacterial activity of IFN1 has physiological significance. Meanwhile, we injected grass carp with the recombinant and purified intact IFN1 protein after bacterial injection, and the result demonstrated a remarkable therapeutic effect. Furthermore, we found that IFN1 expression was remarkably induced in blood cells after bacterial challenge, and prophagocytosis via IFN1 mostly increased in thrombocytes. Then, we isolated peripheral blood thrombocytes by polyclonal Ab of CD41 and stimulated thrombocytes with recombinant IFN1, and the results indicated that immune factors and complement components (especially C3.3) were induced. Unexpectedly, complements demonstrated not only bacteriolysis but also bacterial aggregation. Furthermore, Ab blockades of the three subunits (CRFB1/CRFB2/CRFB5) of the IFN1 receptor or inhibition of STAT1 almost abolished the prophagocytosis via IFN1 and reduced C3.3 and immune factor expression in thrombocytes. Meanwhile, Ab blockade of the complement receptor CR1 greatly attenuated the prophagocytosis of IFN1. In contrast, mouse IFN-β did not show the promotion of antibacterial activity. These results clarify the prophagocytosis and immune regulation pathways of IFN1 in antibacterial immunity in teleosts. This study reveals the antibacterial mechanisms of type I IFNs invivo and inspires functional studies of IFN in bacterial infections.

Hematogenesis Adaptation to Long-Term Hypoxia Acclimation in Zebrafish (Danio rerio)

When fish live in the wild or are cultured artificially, they will inevitably suffer from hypoxia. At the same time, blood physiological indexes represent the physiological state of fish. In order to study the effect of long-term hypoxia acclimation on fish hematogenesis, we cultured zebrafish embryos into adulthood in a hypoxia incubator (1.5 ± 0.2 mg/L). Then we compared the hematological parameters of zebrafish cultured in normoxia and hypoxia conditions. Transcriptome sequencing analysis of the main hematopoietic tissue, the head kidney, was also compared between the two groups. Results showed that the number of erythrocytes increased significantly in the long-term hypoxia acclimated group, while the size of several cell types, such as red blood cells, eosinophils, basophils, small lymphocytes and thrombocytes, decreased significantly. The transcriptomic comparisons revealed that there were 6475 differentially expressed genes (DEGs) between the two groups. A Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that hematopoiesis and cell proliferation signaling were the most significantly enriched pathways in the head kidney of hypoxia acclimated zebrafish. In addition, many genes involved in the hematopoietic process showed significantly higher levels of expression in the hypoxia acclimated zebrafish, when compared to the normoxia zebrafish. When considered together, these data allowed us to conclude that long-term hypoxia can promote the hematopoiesis process and cell proliferation signaling in the zebrafish head kidney, which resulted in higher red blood cell production. Higher numbers of red blood cells allow for better adaptation to the hypoxic environment. In conclusion, this study provides a basis for the in-depth understanding of the effects of hypoxia on hematogenesis in fish species.

Primary care coverage and individual health: evidence from a likelihood\xa0model using biomarkers in Brazil

BackgroundAlthough the use of biomarkers to assess health outcomes has recently gained momentum, literature is still scarce for low- to middle-income countries. This paper explores the relationship between primary care coverage and individual health in Brazil using a dataset of blood-based biomarkers collected by the Brazilian National Health Survey. Both survey data and laboratory results were crossed with coverage data from the Family Health Strategy (ESF) program, the most important primary care program in Brazil; the coverage measures aim to capture both direct (household) and indirect (spill-over) effects.MethodsThe empirical strategy used a probit model to estimate the relationship between ESF program coverage and the likelihood of abnormal biomarker levels while controlling for a rich set of individual and household characteristics based on data from the national survey.ResultsHousehold ESF coverage was associated with a lower likelihood of abnormal results for biomarkers related to anemia (marginal effect between − 2.16 and − 2.18 percentage points), kidney failure (between − 1.01 and − 1.19 p.p.), and arterial hypertension (between − 1.48 and − 1.64 p.p). The likelihood of abnormal levels of white blood cells and thrombocytes was negatively related to primary care coverage (marginal effect between − 1.8 and − 2 p.p.). The spillover effects were relevant for kidney failure and arterial hypertension, depending on the regional level. Although not sensitive to household coverage, diabetes mellitus was negatively associated with the state supply of primary care, and abnormal cholesterol levels did not present any relationship with ESF program coverage.ConclusionsThe presence of spillover effects of ESF program coverage regarding these conditions reveals that the strengthening of primary care by increasing the household registration and the regional density of ESF teams is an efficient strategy to address important comorbidities.

Influence of the Herbal Preparation "Licorice Oil" on the State of Hematopoiesis in Rats Under Ionizing Irradiation

The radioprotective effect of the herbal preparation "Licorice oil" on the hematopoietic system and oxidative stress was studied. The experiment was carried out on 30 male Wistar rats, divided into 3 groups. The first group is control group, the second is irradiation with 6Gy, third group - a week before irradiation and two weeks after, received "Licorice oil" intragastrically at a dose of 2.5 ml/kg of body weight. Gamma irradiation significantly reduced the number of erythrocytes, hemoglobin, hematocrit, leukocytes, thrombocytes in peripheral blood and bone marrow cellularity. The frequency of micronuclei in polychromatophilic erythrocytes of the bone marrow has significantly increased. The level of lipid peroxidation in the blood increased against the background of a significant decrease in the activity of antioxidant enzymes. The introduction of "Licorice oil" for 21 days provided a protective effect. In application of "Licorice oil", there was an increase in the number of cellular elements in the peripheral blood and against the background of a decrease in the frequency of micronuclei in the bone marrow. The activity of antioxidant enzymes in blood plasma increases against the background of a decrease in the amount of peroxidation products. The herbal preparation "Licorice oil" exhibits antioxidant activity, reduces genotoxicity and cytotoxicity under gamma irradiation. "Licorice oil" can be used to prevent radiation damage.

Application of a Four-Tier Diagnosis Test for Evaluating Student's Misconception about Blood Classification

Background: A misconception is a form of error in scientific knowledge that is believed to be accurate. As a result, someone who has a misconception of something will reject scientific truth and will hinder the assimilation of new knowledge. The purpose of this study was to identify misconceptions that occur in students regarding the topic of blood classification. Methods: The survey method research was conducted on 97 students scattered in Malang and Tulungagung, Indonesia. The data analysis technique uses coding for categorization into four categories: conceptual understanding, partial understanding, misconception, and not understanding the concept. Results: The results showed that misconceptions occurred in all sub-topics of blood classification. Misconceptions were found in the concept of blood plasma (2.1%), white blood cells (6.2%), thrombocytes (17.5%), and blood classification system (8.2%). Misconceptions occur due to several misunderstandings of students such as red blood cells that play a role in transporting nutrients, haemoglobin only binds oxygen, and difficulty distinguishing between antigens and antibodies in determining blood groups, blood donation, and the impact arising from donations. Conclusion: The existence of misconceptions causes students' mastery of blood classification material to be hampered.&#x0D; &#x0D; &#x0D; &#x0D;

Assessment of Appropriateness of Cardiovascular Preventive Medication in Adults ≥ 75 years

Physicians are confronted with dilemma’s on cardiovascular preventive medication for older people with complex health problems. With accumulation of diseases, limitations and shortening life expectancy, questions arise for whom cardiovascular preventive treatment is still appropriate (the expected benefits of treatment exceeds the negative consequences by a sufficiently wide margin resulting that treatment is worth doing). There is a need for more guidance in clinical situations. With the RAND/UCLA appropriateness method (RUAM), we investigated the appropriateness of cardiovascular preventive medication in adults ≥ 75 years. The RUAM is a systematic, formalized method, to combine available scientific evidence with the collective judgment of experts. Fourteen interdisciplinary panelists (9 physicians representing 6 medical disciplines, 1 medical ethics expert, 1 pharmacist and 3 older adults [lay man]), discussed and rated the appropriateness of starting and stopping of three medication groups (cholesterol lowering, blood pressure lowering and thrombocyte aggregation inhibitors) for different clinical scenarios (combinations of cardiovascular history, systolic blood pressure, complexity of health problems, age, life-expectancy, side-effects). Depending on the medication group, different patterns of appropriateness judgments across the clinical scenarios were found. In general, absence of cardiovascular disease, presence of complex health problems, a short life-expectancy or hindering side-effects were important factors to judge cardiovascular preventive medication as inappropriate. Results were summarized into colored reading maps. These findings can offer more guidance in clinical decision making about cardiovascular preventive treatment for adults ≥ 75 years.

ПОКАЗАТЕЛИ ПЕРИФЕРИЧЕСКОЙ КРОВИ КАК ПРЕДИКТОРЫ ЭФФЕКТИВНОСТИ ВЛИЯНИЯ ГИДРОФИЛЬНОЙ ФРАКЦИИ ПЧЕЛИНОЙ ОБНОЖКИ НА ОРГАНИЗМ КРЫС ПРИ ИММОБИЛИЗАЦИОННОМ СТРЕССЕ

The novelty of the research involves studying the influence of the innovative product such as hydrophilic factions of bee pollen on an organism in a view of a classical predictor that is an indica-tor of peripheral blood. It is proved that multi-component structure helps the product optimize blood cell structure of the rats from the test group under stress. It equalizes it to the level of the intact an-imals. With increasing stress caused by immobilization the changes of hematopoiesis of the rats were noticed such as erythrocyte, leukocytosis with simultaneous replication, increasing number of thrombocytes in blood. There were no such changes after using hydrophilic factions of bee pollen in the test group, like it was observed in the test group after prolonged usage of the products. It is pos-sible that they were so discreet and didn't influence the indicators under the research. Mainly it is caused by highly-expressed antioxidant quality, prevention of making active form of oxygen, ap-pearing with increasing stress and formulation of basis for restoring homeostatic mechanisms in an organism. The practical interest is connected with famous general biological mechanism of stress. So there are some opportunities that the results of the research can be projected on both humans and animals.

A Randomized Controlled Trial of Long-Term (R)-α-Lipoic Acid Supplementation Promotes Weight Loss in Overweight or Obese Adults without Altering Baseline Elevated Plasma Triglyceride Concentrations

α-Lipoic acid (LA) is a dietary supplement for maintaining energy balance, but well-controlled clinical trials in otherwise healthy, overweight adults using LA supplementation are lacking. The primary objective was to evaluate whether LA supplementation decreases elevated plasma triglycerides in overweight or obese adults. Secondary aims examined if LA promotes weight loss and improves oxidative stress and inflammation. Overweight adults [n=81; 57% women; 21-60 y old; BMI (in kg/m2)≥25] with elevated plasma triglycerides ≥100mg/dL were enrolled in a 24-wk, randomized, double-blind, controlled trial, assigned to either (R)-α-lipoic acid (R-LA; 600mg/d) or matching placebo, and advised not to change their diet or physical activity. Linear models were used to evaluate treatment effects from baseline for primary and secondary endpoints. R-LA did not decrease triglyceride concentrations, but individuals on R-LA had a greater reduction in BMI at 24 wk than the placebo group (-0.8; P=0.04). The effect of R-LA on BMI was correlated to changes in plasma triglycerides (r = +0.50, P=0.004). Improvement in body weight was greater at 24 wk in R-LA subgroups than in placebo subgroups. Women and obese participants (BMI≥35) showed greater weight loss (-5.0% and -4.8%, respectively; both P<0.001) and loss of body fat (-9.4% and -8.6%, respectively; both P<0.005). Antioxidant gene expression in mononuclear cells at 24 wk was greater in the R-LA group (Heme oxygenase 1 [HMOX1] : +22%; P=0.02) than in placebo. Less urinary F2-isoprostanes (-25%; P=0.005), blood leukocytes (-10.1%; P=0.01), blood thrombocytes (-5.1%; P=0.03), and ICAM-1 (-7.4%; P=0.04) at 24 wk were also observed in the R-LA group than in placebo. Long-term LA supplementation results in BMI loss, greater antioxidant enzyme synthesis, and less potential for inflammation in overweight adults. Improved cellular bioenergetics is also evident in some individuals given R-LA.This trial was registered at clinicaltrials.gov as NCT00765310.

Effects of Artemisia annua alcohol extract on physiological and innate immunity of Nile tilapia (Oreochromis niloticus) to improve health status

Plants are a potential source of active molecules and are environmentally safer and cheaper than synthetic antibiotics. Bioactive compounds of Artemisia annua have shown pharmacological activities and are used globally as a supplement. The present study tested whether dietary supplementation with alcohol extract of the plant A. annua (ae-Aa; patent BR10201902707) improves the health status of juvenile Nile tilapia and increases resistance to diseases when fish are challenged with the bacteria Aeromonas hydrophila. The experimental design was completely randomized with four experimental groups (0.0, 0.1, 0.25, and 0.5% ae-Aa in the diets) with five repetitions (12 fish per repetition/experimental unit). We assessed serum glucose and cortisol levels in plasma, leukocyte respiratory activity, total plasma protein, serum lysozyme levels, as well as the number of circulating red blood cells and fish leukocytes at the end of the 30 days of feeding (phase I) and 24h after exposure to bacteria (phase II). The levels of lipid peroxidation, catalase activity and glutathione S-transferase of fish were also analyzed. The supplementation of 0.5% of ae-Aa was sufficient to increase the respiratory burst of leukocyte and lysozyme activity, total plasma protein, blood thrombocytes, neutrophils and monocytes after bacterial challenge (P < 0.05), and minimized stress response with decreases in plasmatic glucose and cortisol, and reduction in lipid peroxidation levels (P < 0.05). Results of the present study suggest that ae-Aa as a dietary supplement has benefits, including supplementation with 0.5% A. annua extract for 30 days to minimize the stress response and modulate innate immunity in Nile tilapia, providing fish with greater resistance and disease protection.

P1669PROGNOSTIC IMPORTANCE OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IN RENAL TRANSPLANTATION

Abstract Background and Aims The prevalence of end-stage renal disease (ESRD) has increased ten times higher in the past twenty years, where renal replacement therapy (dialysis or kidney transplantation (KTx)) is the sole life-saving treatment. KTx is the preferred option as it is associated with improved survival and quality of life as well. Delayed graft function (DGF) is one of the main problems affecting long-term kidney survival. Brain-derived neurotrophic factor (BDNF) signalling pathways play pivotal role in mitigating cerebral ischemia/reperfusion injury (IRI), however the relation of BDNF and IRI in KTx is unknown. The aim of our human clinical study was to explore the relationship between serum BDNF concentration, BDNF gene polymorphism and renal graft function after KTx. Method Study characteristics: We enrolled 59 ESRD patients with average age of 54.8±12 years who received KTx. Proportion of male patients was 57%. Average cold ischemic time was 927±310 min, warm ischemic time was 54.5±39 min. DGF occurred in 5 cases. Baseline triple immunosuppression therapy consisted of tacrolimus, mycophenolate or everolimus, and prednisolone. Until now, 44 patients completed the 2 years follow-up. For a comparable control group, we collected blood samples from 79 healthy volunteers with average age of 53.9±16 years and with male gender proportion of 52%. Serum BDNF, creatinine, blood urea nitrogen, haemoglobin, blood glucose level and thrombocyte numbers were measured before KTx and 1 week, 1-, 3-, 6 months, and 1-, 2 years after transplantation, as well as in controls. GFR was estimated based on the CKD-EPI formula. BDNF Val66Met polymorphism was determined by PCR-RFLP. Results There was no difference in genotype or allele distribution between any of the groups, and no correlation could be observed between serum BDNF and different genotypes either. Serum BDNF level was lower in ESRD patients than healthy controls (p=0.03). There was a weak correlation and marginal significance (p=0.056) between eGFR and serum BDNF level in controls, while in KTx recipients this correlation reached higher significance (p=0.01). Above median BDNF values at 1 month after KTx were predictive for better graft function during the 2 observed years. Conclusion Our preliminary human study proposes that BDNF could be a novel biomarker of posttransplant graft function, however further clinical studies with significantly larger population are definitely needed to confirm these results.

Characteristics of Hematopoiesis in Follicular Lymphoma Patients

Aim. To assess hematopoiesis in follicular lymphoma (FL) patients at different disease stages with different morphologic structures of tumor and bone marrow microenvironment. Materials &amp; Methods. The trial included 152 FL patients treated from 2006 to 2016. In all of them the diagnosis was based on immunohistochemical analysis of extramedullar tumor as well as the analysis of bone marrow aspirates and core biopsy samples. In cases of bone marrow lesions (n = 33) a detailed morphoimmunophenotypic evaluation of tumor cells was carried out by means of flow cytometry, and lymphocyte subset panel evaluation was performed. Results. Anemia, thrombocytopenia, and monocytosis in blood of FL patients are not associated with bone marrow lesions. In the absence of signs of these lesions anemia was detected in 23 (19 %) patients, thrombocytopenia was identified in 8 (7 %) patients, and 11 (9.1 %) patients showed monocytosis. Among patients with bone marrow lesions 9 (27.2 %) anemia, 11 (33.8 %) thrombocytopenia, and 7 (21 %) monocytosis cases were reported. Depth of cytopenia was determined by the degree of bone marrow tumor infiltration. Based on lymphocyte subset panel evaluation the following types of tumor cells in bone marrow aspirates were characterized: elements with blastic structure of nuclear chromatin, atypical lymphoid cells, and those similar to normal lymphocytes. Immunophenotypic heterogeneity of tumor cells in bone marrow was demonstrated. The trial showed that hemoglobin level, the count of blood thrombocytes and monocytes as well as the count of bone marrow T-cells are not associated with types of tumor cells. Conclusion. Arrest of hematopoiesis and increasing number of monocytes in blood correlate with the degree of bone marrow tumor infiltration and are not affected by morphoimmunological characteristics of FL tumor cells.

P6141Associations between atmospheric parameters and haemostatic factors: a case control study

Abstract Background One of the biggest health challenges in the 21st century is global climate change. The health effect of climate change is partly mediated through atmospheric parameters. There is a growing concern that atmospheric parameters might increase cardiovascular (CV) morbidity. Increased levels of haemostatic factors are predictors of CV events. The associations between CV diseases and atmospheric parameters have been widely reported, however there are few studies of atmospheric parameters' effects on markers of haemostasis. Purpose We examined the possible association between atmospheric parameters and several haemostatic markers. Methods The study consisted of 3800 hospitalized patients with acute CV diseases (ACVDs) and 260 healthy blood donors. We examined the relationship of haemoglobin (Hgb), white blood cells (WBC), thrombocytes (THR) and local atmospheric parameter conditions (temperature, atmospheric pressure, humidity, wind speed, atmospheric front) on a day-to-day basis in a 5-year period (2009–2013) using a General Additive Model with cubic splines of covariates, regularized by a ridge penalty, and employing generalized cross validation. Atmospheric parameters were allowed to have a lagged effect by up to 21 days. Results Among blood donors, the average of daily temperature lagged by 8–14 days had a significant effect on Hgb, exhibiting a U-shaped relationship, where higher Hgb values were associated with extremities of the observed temperature interval. For ACVD patients, all examined blood test variables has a significant association with at least some of the atmospheric parameters. Hgb was shown to have a negative linear relationship with mean daily humidity, and the average of daily temperature lagged by 15–21 days, while the average of daily temperature variation lagged by 2–7 days had highly non-linear effect. The relative strength of the association with Hgb was largest for daily temperature variation. WBC values had a slightly non-linear positive relationship with atmospheric pressure lagged by 1 day, with WBC being significantly increased above 1030 hPa. THR values decreased linearly with an increase in mean daily temperature averaged for days lagged 15–21. Atmospheric pressure lagged by 1 day also had a significant effect on THR, with a positive linear effect under 1010 and over 1020 hPa but no effect between. The relative effect of atmospheric pressure on THR was twice as large compared to temperature. Conclusions Our study showed that exposure to certain atmospheric parameters is associated with significant changes in haemostatic marker levels. In the context of global climate change, the importance of focusing on atmospheric parameters as minor CV risk factor is substantially growing. A better understanding of the fluctuation of the examined markers, in light of atmospheric parameters, appears to be of particular importance for future studies and could help establish new CV prevention strategies.

The aqueous extract of the root bark of &lt;i&gt;Psorospermum febrifugum&lt;/i&gt; Spach effectively corrects anaemia. Experimental study on &lt;i&gt;Wistar&lt;/i&gt; rats

Objectives: Psorospermum febrifugum Spach. (Clusiaceae) is a medicinal plant found in Africa. It root bark was used in Benin to treat anaemia. This work aimed to study the therapeutic efficacy of that root bark extract on anaemia treatment. Methodology and Results: Wistar Rats were anaemic by intraperitoneal injection of phenylhydrazine chloridrate. The rats were then fed either with Psorospermum febrifugum Spach root bark extract at 200 or 300 mg / kg weight / day, or with vitafer® as the reference drug. Non-anaemic rats served as controls. The blood count and osmotic resistance of red blood cells carried out on blood samples of rats at different days (D 0 , D 2 , D 7 , J 10 and D15) showed a correction of anaemia with the extract in two weeks. The extract screening revealed saponosides, reducing sugars, polyphenols, flavonoids, tannins, proteins, anthocyanins and alkaloids. Conclusion and application of results: The extract completely corrected anaemia in two weeks by increased stimulation of hemoglobin synthesis and the production of young red blood cells released into the blood. Its effect seems dose-dependent and at the 300 mg / Kg dose was even more effective than the vitafer. This remarkable efficiency would certainly be linked to its chemical composition, whose elements would act in an isolated or synergistic way to stimulate haematopoiesis.. In addition, the extract had no effect on blood thrombocytes suggesting a certain specificity of action on the erythrocyte line. Psorospermum febrifugum Spach root bark had an excellent therapeutic efficacy and could be a good candidate for transformation into improved traditional medicines (ITM) for anaemia treatment, after acute and chronic toxicity tests and appropriate clinical trials. Keywords: Psorospermum febrifugum Spach , anaemia, osmotic resistance

Diphenyl diselenide modulates splenic purinergic signaling in silver catfish fed diets contaminated with fumonisin B1: An attempt to improve immune and hemostatic responses

The aim of this study was to determine whether purinergic signaling is a pathway associated with fumonisin B1 (FB1)-induced impairment of immune and hemostatic responses. We also determined whether dietary supplementation with diphenyl diselenide (Ph2Se2) prevents or reduces these effects. Splenic nucleoside triphosphate diphosphohydrolase (NTPDase) activity for adenosine triphosphate (ATP) and adenosine diphosphate (ADP) as substrates and total blood thrombocytes counts were significant lower in silver catfish fed with FB1-contaminated diets than in fish fed with a basal diet, while splenic adenosine deaminase (ADA) activity and metabolites of nitric oxide (NOx) levels were significant higher. Also, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significant lower in silver catfish fed with FB1-contaminated diets than in fish fed with a basal diet. Dietary supplementation with 3 mg Ph2Se2/kg of feed effectively modulated splenic NTPDase (ATP as substrate), ADA, GPx and SOD activities, as well as NOx levels, and was partially effective in the modulation of spleen NTPDase activity (ADP as substrate) and total blood thrombocytes count. These data suggest that splenic purinergic signaling of silver catfish fed with FB1-contaminated diets generates a pro-inflammatory profile that contributes to impairment of immune and inflammatory responses, via reduction of splenic ATP hydrolysis followed possible ATP accumulation in the extracellular environment. Reduction of ADP hydrolysis associated with possible accumulation in the extracellular environment can be a pathophysiological response that restricts the hemorrhagic process elicited by FB1 intoxication. Supplementation with Ph2Se2 effectively modulated splenic enzymes associated with control of extracellular nucleotides (except ADP; that was partially modulated) and nucleosides, thereby limiting inflammatory and hemorrhagic processes.

Disturbances of the ultrastructure of endotheliocytes of sinusoid hemocapillaries of the liver and the hemostasis system in the acute period of cranial injury

Purpose of the study : the role of endothelial cells of the sinusoidal capillaries of the liver in the pathogenesis of the vascular-platelet and coagulation units of the hemostasis system to assess the acute period of traumatic brain injury. Materials and methods . Work is a clinical experimental study. TBI was simulated in animals and the number of blood thrombocytes was determined at 1, 3, 7, 12 days after the alteration. Also on the 3 rd , 7 th and 12 th day after the application of TBI, structural changes in sinusoidal hemocapillaries (SH) were studied in ultrathin sections of the liver. The study of patients was conducted at the Semashko Hospital. Hemostasis systems were evaluated in patients by the indicators: activated partial thromboplastin time, prothrombin and thrombin time, prothrombin index, fibrinogen, antithrombin III, soluble fibrin-monomer complexes, plasma fibrinolytic activity, platelet count in 1-e (on admission) and 3, 7 and 10 days of treating patients. Results . The results of electron microscopic examination of the liver SH and intravascular changes revealed that pronounced damage to endotheliocytes and microcirculation disorders were determined in animals on the 3 rd day of the post-traumatic period. These changes were aggravated on the 7 th day after the injury. Partial restoration of blood flow and endothelium with the preservation of areas of its swelling and edema occurred 12 days after TBI. The number of platelets significantly decreased with a maximum change in the index by day 7 and recovered to the values of intact rats on the 12 th day after TBI. The number of platelets decreased and coagulation hemostasis increased in patients with severe concurrent TBI on the 3 rd and 7 th day after injury compared to the reference intervals of the studied hemostasis parameters. Conclusion . In this way, a combination of non-specific systemic disorders of the structural and functional integrity of the vascular endothelium and activation of the blood coagulation system is observed in the acute period of severe TBI, which dictates the need for targeted prevention and correction of the above disorders in the complex treatment of patients.

An overlooked fact: thrombocytopenia following bioprosthetic aortic valve replacement.

IntroductionThrombocytopenia observed after bioprosthetic aortic valve replacement has remained a puzzle to solve.AimTo analyze thrombocytopenia occurring after bioprosthetic aortic valve replacement by comparison to mechanical aortic valve replacement and coronary artery bypass grafting procedures.Material and methodsThe study was conducted retrospectively on a total of 297 patients who underwent bioprosthetic aortic valve replacement, mechanical prosthetic aortic valve replacement and coronary artery bypass grafting at the cardiovascular surgery department of our clinical center between January 2013 and September 2017. Preoperative and postoperative first 14-day thrombocyte levels of the patients were analyzed.ResultsThe postoperative blood thrombocyte level decrease was found to be more significant in patients who underwent bioprosthetic aortic valve replacement than in patients who underwent mechanical aortic valve replacement and coronary artery bypass grafting (p < 0.01). There was also a statistically significant difference in the time to reach the lowest postoperative platelet levels according to type of surgery (p = 0.001; p < 0.01).ConclusionsWhen compared to the patients who underwent coronary artery bypass grafting and mechanical prosthetic aortic valve replacement, postoperative thrombocytopenia was found to be more severe in patients who underwent bioprosthetic aortic valve replacement, with a dramatic decrease in thrombocyte count being observed on the postoperative second day. It was found that the thrombocytopenia recovers without causing any problem. We think that the shear forces may play a role in this recovery through washout of chemicals responsible for thrombocytopenia from the glutaraldehyde treated bioprostheses.

Both Granulocytic and Non-Granulocytic Blood Cells Are Affected in Patients with Severe Congenital Neutropenia and Their Non-Neutropenic Family Members: An Evaluation of Morphology, Function, and Cell Death

Objective:To examine granulocytic and non-granulocytic cells in children with severe congenital neutropenia (SCN) and their non-neutropenic parents.Materials and Methods:Fifteen patients with SCN and 21 non-neutropenic parents were evaluated for a) CD95, CD95 ligand, annexin V, propidium iodide, cell cycle, and lymphocyte subsets by flow cytometry; b) rapid cell senescence (of leukocytes) by senescence-associated β-galactosidase stain; c) aggregation tests by aggregometer; d) in vitro bleeding time by PFA-100 instrument; e) mepacrine-labeled dense granule number of thrombocytes by fluorescence microscope; and f) hematomorphology by light and electron microscope. HAX1, ELANE, G6PC3, CSF3R, and JAGN1 mutations associated with SCN were studied in patients and several parents.Results:Significant increase in apoptosis and secondary necrosis in monocytes, lymphocytes, and granulocytes of the patients and parents was detected, irrespective of the mutation type. CD95 and CD95 ligand results implied that apoptosis was non-CD95-mediated. Leukocytes of 25%, 12.5%, and 0% of patients, parents, and controls showed rapid cell senescence. The cell cycle analysis testable in four cases showed G1 arrest and apoptosis in lymphocytes of three. The patients had HAX1 (n=6), ELANE (n=2), G6PC3 (n=2), and unidentified (n=5) mutations. The CD3, CD4, and NK lymphocytes were below normal levels in 16.6%, 8.3%, and 36.4% of the patients and in 0%, 0%, and 15.4% of the parents (controls: 0%, 0%, 5.6%). The thrombocytes aggregated at low rates, dense granule number/thrombocyte ratio was low, and in vitro bleeding time was prolonged in 37.5%-66.6% of patients and 33.3%-63.2% of parents (vs. 0% in controls). Under electron and/or light microscope, the neutrophils, monocytes, lymphocytes, and thrombocytes in the peripheral blood of both patients and parents were dysplastic and the bone marrow of patients revealed increased phagocytic activity, dysmegakaryopoiesis, and necrotic and apoptotic cells. Ultrastructurally, thrombocyte adhesion, aggregation, and release were inadequate.Conclusion:In cases of SCN, patients’ pluripotent hematopoietic stem cells and their non-neutropenic parents are both affected irrespective of the genetic defect.

Brain Barrier Disruption and Region-Specific Neuronal Degeneration during Necrotizing Enterocolitis in Preterm Pigs

Necrotizing enterocolitis (NEC) increases the risk of brain injury and impaired neurodevelopment. Rapid brain maturation prior to birth may explain why preterm brains are particularly vulnerable to serious infections. Using pigs as models, we hypothesized that preterm birth was associated with altered blood-cerebrospinal fluid (CSF) barrier (BCSFB) function and cerebral structural deficits, and that NEC was associated with systemic inflammation, BCSFB disruption, and neuroinflammation. First, cesarean-delivered preterm and term pigs (n = 43–44) were euthanized at birth to investigate BCSFB function and markers of brain structural maturation, or on day 5 to measure markers of blood-brain barrier maturation in the hippocampus and striatum (experiment 1). Next, preterm pigs (n = 162) were fed increasing volumes of infant formula to assess NEC lesions, systemic inflammation, BCSFB permeability, cerebral histopathology, hippocampal micro­glial density, and cytokine levels on day 5 (experiments 2 and 3). In experiment 1, preterm newborns had increased CSF-plasma ratios of albumin and raffinose, reduced CSF glucose levels, as well as increased cerebral hydration and reduced white matter myelination compared with term animals. We observed lower hippocampal (but not striatal) perivascular astrocyte coverage for the first 5 days after preterm birth, accompanied by altered cell junction protein levels. In experiments 2 and– 3, piglets with severe NEC lesions showed reduced blood thrombocytes and increased plasma C-reactive protein and interleukin-6 levels. NEC was associated with increased CSF-plasma albumin and raffinose ratios, reduced CSF leukocyte numbers, and increased cerebral hydration. In the hippocampus, NEC was associated with pyramidal neuron loss and increased interleukin-6 levels. In the short term, NEC did not affect cerebral myelination or microglia density. In conclusion, altered BCSFB properties and brain structural deficits were observed in pigs after preterm birth. Acute gastrointestinal NEC lesions were associated with systemic inflammation, increased BCSFB permeability and region-specific neuronal damage. The results demonstrate the importance of early interventions against NEC to prevent brain injury in preterm infants.

The possibility of applying historical concentration of thrombocytes in examination of donors.

The historical concentration of thrombocytes is an average of three subsequent concentrations of thrombocytes before donation (during two years) in blood of donors. The evaluation was applied to characteristics of historical concentration of thrombocytes concerning concentration of thrombocytes in blood of 99 regular donors before every donation of apheresis thrombocytes. It is established that less than 20% dispersion between historical concentration of thrombocytes and value obtained in day of donation is observed in 96% of regular donors. The average concentration of thrombocytes before three subsequent donations of apparatus apheresis in males is statistically higher than in females: 235,2±6,7 and 222,1±11,0 (× 109 cells/l) correspondingly (p < 0,05). The concentration of thrombocytes has no relationship with intervals between donations. There is no statistically significant differences between historical concentration of thrombocytes and concentration of thrombocytes before fourth donation and in general and between separate donations both in males and females. It is appropriate to analyze quality of concentrates of thrombocytes received through program accounting historical concentration of thrombocytes.