Leukocyte Plasma Research Articles

Dietary ellagic acid blocks inflammation-associated atherosclerotic plaque formation in cholesterol-fed apoE-deficient mice.

Atherosclerosis particularly due to high circulating level of low-density lipoprotein is a major cause of cardiovascular diseases. Ellagic acid is a natural polyphenolic compound rich in pomegranates and berries. Our previous study showed that ellagic acid improved functionality of reverse cholesterol transport in murine model of atherosclerosis. The aim of this study is to investigate whether ellagic acid inhibited inflammation-associated atherosclerotic plaque formation in cholesterol-fed apolipoprotein E (apoE)-knockout (KO) mice. Wild type mice and apoE-KO mice were fed a cholesterol-rich Paigen diet for 10 weeks to induce severe atherosclerosis. Concurrently, 10 mg/kg ellagic acid was orally administered to the apoE-KO mice. Plaque lesion formation and lipid deposition were examined by staining with hematoxylin and eosin, Sudan IV and oil red O. The plasma leukocyte profile of cholesterol-fed mice was not altered by apoE deficiency. Oral administration of ellagic acid attenuated plaque lesion formation and lipid deposition in the aorta tree of apoE-KO mice. Ellagic acid substantially reduced plasma levels of soluble vascular cell adhesion molecule and interferon-γ in Paigen diet-fed apoE-KO mice. When 10 mg/kg ellagic acid was administered to cholesterol-fed apoE-KO mice, the levels of CD68 and MCP-1 were strongly reduced in aorta vessels. The protein expression level of nitric oxide synthase-2 (NOS2) in the aorta was highly enhanced by supplementation of ellagic acid to apoE-KO mice, but the expression level of heme oxygenase-1 (HO-1) in the aorta was reduced. Furthermore, ellagic acid diminished the increased aorta expression of the inflammatory adhesion molecules in cholesterol-fed apoE-KO mice. The treatment of ellagic acid inhibited the scavenger receptor-B1 expression in the aorta of apoE-KO mice, while the cholesterol efflux-related transporters were not significantly changed. These results suggest that ellagic acid may be an atheroprotective compound by attenuating apoE deficiency-induced vascular inflammation and reducing atherosclerotic plaque lesion formation.

The effect of two platelet-rich plasma aspiration techniques on plasma cellular concentrations using a double syringe gravitational centrifugation system.

To identify which aspiration technique increased plasma platelet concentration and which technique minimized plasma leukocyte and erythrocyte concentrations using a gravitational double-syringe platelet rich plasma (PRP) system. Controlled laboratory study. Thirty adult dogs. Whole blood was collected into two autologous conditioned plasma (ACP) syringes and an ethylenediaminetetraacetic acid (EDTA tube) (control samples). The ACP syringes were centrifuged for 5 min at 1500 rpm. The proximal 2 mL of plasma from one ACP syringe was deposited in an EDTA tube (preflash samples). Plasma from the second ACP syringe was withdrawn until the buffy coat was pierced, producing a "flash" of red blood cells, agitated and deposited into an EDTA tube (flash samples). Complete blood counts were performed. Mean plasma platelet concentrations of the control, preflash, and flash samples were 2.4 × 105/dL, 3.3 × 105/dL and 4.1 × 105/dL, respectively. The mean platelet concentration of the flash samples was 7.9 × 104/dL higher than the preflash samples (p = .005). The mean platelet concentration was lower in the control samples than the preflash (p = .002) and flash (p < .0001) samples. The median plasma leukocyte concentration of the preflash samples (0/dL) was lower than in the flash samples (2.4 × 103/dL) (p = .001). The median plasma hematocrit value of the preflash samples (0%) was lower than in the flash samples (1.0%) (p = .002). The flash method is not necessary to produce a PRP sample. Both methods produced PRP. However, clinicians should avoid aspirating the buffy coat when processing PRP for therapies where leukocytes and erythrocytes are contraindicated.

Preventive and Therapeutic Effects of Total Flavonoids of Willow Buds on Acute Pancreatitis in Mice

To explore the preventive and therapeutic effects of total flavonoids of willow buds on acute pancreatitis is the objective of the study. The willow buds of Salix babylonica L. (~1 cm) were harvested from Shenyang Institute of Technology between March 2020 to April 2020. Ultrasound-assisted extraction of total flavonoids of willow buds was conducted and an L-arginine-induced acute pancreatitis mouse model was established. The content of naringenin was determined by Agilent 1100 high-performance liquid chromatography. Serum biochemical test, serum oxidation test and plasma leukocyte content was determined and HE staining was performed. The protective and therapeutic effects of total flavonoids of willow buds on acute pancreatitis in mice were investigated. Total flavonoids of willow buds effectively decreased the activity of serum amylase, lipase and malondialdehyde, reduced plasma white blood cells and increased serum calcium content and superoxide dismutase activity in acute pancreatitis mice; total flavonoids of willow buds alleviated the pathological injury of the pancreas in acute pancreatitis mice. In conclusion, total flavonoids of willow buds may reduce pancreatic injury via reducing the inflammatory response and increasing the activity of antioxidant enzymes, which is a promising natural anti-acute pancreatitis drug.

Abstract 532: Circulating tumor DNA and outcomes with lutetium-PSMA in advanced prostate cancer: Preliminary results from an Australian study

Abstract Recent clinical trials have revealed that Lutetium PSMA-617 (Lu-PSMA, a small molecular inhibitor of prostate-specific membrane antigen radiolabelled with the beta emitter 177-Lutetium) significantly improves clinical outcomes including overall survival for patients with metastatic castration resistant prostate cancer (mCRPC). Nevertheless, owing to the highly heterogeneous nature of mCRPC, responses to Lu-PSMA therapy can be variable, with resistance inevitable. Currently, there are no existing data on the genomic landscape of mCRPC in the context of this therapy, with the identification of biomarkers linked to outcomes with Lu-PSMA being a critical unmet need in order to select patients most likely to benefit from treatment and to dissect mechanisms of resistance. mCRPC patients with high PSMA expression on positron emission tomography scans and prior treatment with docetaxel chemotherapy and an androgen receptor pathway inhibitor were enrolled on a compassionate access prospective registry at Peter MacCallum Cancer Centre. Patients received Lu-PSMA every 6 weeks for 4-6 cycles and entered into follow-up. Peripheral blood samples were collected for plasma circulating tumour DNA (ctDNA) analysis at baseline and prior to cycle 2 of therapy. A highly-sensitive targeted next-generation sequencing assay was applied to patient plasma and matched leukocytes. Somatic mutations (single nucleotide variants/indels) and copy number alterations from 43 genes were reported, along with the estimated ctDNA fraction. Current patient numbers preclude formal statistical analysis of genomic aberrations, however dynamic changes in ctDNA were assessed in relation to prostate-specific antigen (PSA) response and clinical and/or radiographic progression. In total, 19 plasma samples were analysed from 13 mCRPC patients with a median age of 71. Patients had between one and four prior lines of therapy for mCRPC and had a median time on Lu-PSMA therapy of 3.7 months. ctDNA was detected in 17/19 plasma samples with a median fraction of 26% (range 0-89%). Overall, a PSA response rate of 54% was observed. Interestingly, patients with higher baseline ctDNA fractions were more likely to experience a PSA response to Lu-PSMA therapy (median ctDNA fraction of responders 49% vs 10% in non-responders, p=0.1). Additionally, no PSA responses were seen in patients who did not attain a reduction in plasma ctDNA from baseline to cycle 2. This is the first targeted assessment of serial plasma samples from patients receiving Lu-PSMA therapy. Using a highly sensitive liquid biopsy assay, ctDNA was detectable in almost all samples. In this preliminary analysis, both baseline ctDNA fraction and early dynamic changes in ctDNA fraction appear to have clinical importance in Lu-PSMA therapy. Recruitment to this registry is proceeding rapidly, with further data available by April 2022. Citation Format: Heidi Fettke, Nicole Ng, Christine Hauser, Patricia Bukczynska, Elizabeth Medhurst, Louise Kostos, James Buteau, Jason Steen, Tu Nguyen-Dumont, Michael Hofman, Arun A. Azad. Circulating tumor DNA and outcomes with lutetium-PSMA in advanced prostate cancer: Preliminary results from an Australian study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 532.

Cytokine autoantibodies are stable throughout the haematopoietic stem cell transplantation course and are associated with distinct biomarker and blood cell profiles

Cytokine-specific autoantibodies (c-aAbs) represent an emerging field in endogenous immunodeficiencies, and the immunomodulatory potential of c-aAbs is now well documented. Here, we investigated the hypothesis that c-aAbs affects inflammatory, immunoregulatory and injury-related processes and hence the clinical outcome of haematopoietic stem cell transplantation (HSCT). C-aAbs against IL-1α, IL-6, IL-10, IFNα, IFNγ and GM-CSF were measured in 131 HSCT recipients before and after (days + 7, + 14, + 28) HSCT and tested for associations with 33 different plasma biomarkers, leukocyte subsets, platelets and clinical outcomes, including engraftment, GvHD and infections. We found that c-aAb levels were stable over the course of HSCT, including at high titres, with few individuals seeming to acquire high-titre levels of c-aAbs. Both patients with stable and those with acquired high-titre c-aAb levels displayed significant differences in biomarker concentrations and blood cell counts pre-HSCT and at day 28, and the trajectories of these variables varied over the course of HSCT. No clinical outcomes were associated with high-titre c-aAbs. In this first study of c-aAbs in HSCT patients, we demonstrated that high-titre levels of c-aAb may both persist and emerge in patients over the course of HSCT and may be associated with altered immune biomarkers and cell profiles.

Aldosterone suppresses cardiac mitochondria

Elevated serum aldosterone promotes arterial hypertension, cardiac hypertrophy, and diastolic dysfunction. However, the effect of elevated aldosterone levels on cardiac mitochondria remains unclear. We used primary cultures of mouse cardiomyocytes to determine whether aldosterone has direct effects on cardiomyocyte mitochondria, and aldosterone-infused mice as a preclinical model to evaluate the impact of aldosterone in vivo. We show that aldosterone suppressed mtDNA copy number and SOD2 expression via the mineralocorticoid receptor (MR)-dependent regulation of NADPH oxidase 2 (NOX2) and generation of reactive oxygen species (ROS) in primary mouse cardiomyocytes. Aldosterone suppressed cardiac mitochondria adenosine triphosphate production, which was rescued by N-acetylcysteine. Aldosterone infusion for 4 weeks in mice suppressed the number of cardiac mitochondria, mtDNA copy number, and SOD2 protein expression. MR blockade by eplerenone or the administration of N-acetylcysteine prevented aldosterone-induced cardiac mitochondrial damage in vivo. Similarly, patients with primary aldosteronism had a lower plasma leukocyte mtDNA copy number. Plasma leukocyte mtDNA copy number was positively correlated with 24-hour urinary aldosterone level and left ventricular mass index. In conclusion, aldosterone suppresses cardiac mitochondria in vivo and directly via MR activation of ROS pathways.

Risk Factors for Hospital Mortality in Acute ST-Segment Elevation Coronary Syndrome Complicated by Cardiogenic Shock

The aim. To study the risk factors for hospital mortality in patients with acute coronary syndrome with ST-segment elevation (STEACS) complicated by cardiogenic shock (CS).Materials and methods. A total of 104 patients with STEACS complicated by CS were studied. The follow-up group (group I) included 58 (55,8%) patients who died in hospital (mean age 71,8±7,31 years), the comparison group (group II) – 46 patients, who have been treated and discharged (mean age 59,5±6,18 years). All patients underwent general clinical studies, the level of troponins, lipids, glucose, creatinine in plasma was determined, electrocardiography and echocardiography were performed. Coronary angiography and percutaneous coronary intervention (PCI) were urgently performed. The method of binary logistic regression with the determination of the odds ratio and its 95% confidence interval for each reliable variable was used to identify risk factors for hospital mortality.Results. In group I patients with CS, compared with group II, patients over the age of 70 (32 (55,2%) vs 10 (22,7%), р=0,0004), with concomitant chronic kidney disease (32 (55,2%) vs 9 (19,6%), p=0,0002), postinfarction cardiosclerosis (30 (51,7%) vs 9 (19,6%), р=0,001) and chronic heart failure of III-IV functional class (32 (55,1%) vs 11 (23,9%), p=0,001) were significantly more often observed. Baseline levels of plasma leukocytes, troponin and creatinine were significantly higher in deceased patients with CS. Left ventricular ejection fraction below 40% was observed more often in the follow-up group than in the comparison group (46 (79,3%) vs 27 (58,7%), p=0,022). In group I, compared with group II, there was a higher incidence of three-vessel coronary lesions (36 (75%) vs 12 (26,1%), p=0,0001) and chronic coronary artery occlusion unrelated to STEACS (25 (52,1%) vs 12 (26,1%), р=0,009). The same trend was observed when assessing the average number of stenoses and occlusions of the coronary arteries. PCI was performed in 43 (74,1%) of the deceased and 43 (93,5%) of the surviving STEACS patients with CS (p=0,009). The follow-up group had a higher rate of unsuccessful PCI (30,2%) vs 3 (7%), р=0,001) and performed later than 6 hours after the onset of an angina attack (28 (65,1%) vs 6 (14%), р=0,0001).Summary. Hospital mortality in patients with STEMI complicated by CS was associated with the presence left ventricular ejection fraction less than 40%, three-vessel coronary lesion and performing PCI later than 6 hours from the beginning of the pain attack.

Plasma Leukocyte Cell-Derived Chemotaxin 2 (LECT2) as a Risk Factor of Coronary Artery Disease: A Cross-Sectional Study.

Many studies have shown that leukocyte cell-derived chemotaxin 2 (LECT2) is associated with metabolic disorders, which is a risk factor of arteriosclerosis. We assessed the level of LECT-2 in patients with coronary artery disease (CAD) and its severity and prognosis. We selected 666 participants who underwent coronary angiography in our hospital and included patients with non-CAD, patients with stable angina pectoris (SAP), patients with unstable angina (UA), patients with non-ST-segment elevation myocardial infarction (NSTEMI) and patients with ST-segment elevation myocardial infarction (STEMI). The serum level of LECT-2 was higher in patients with CAD than in patients with non-CAD and was an independent predictor for CAD. Subgroup analysis showed that compared with the SAP group, the UA, NSTEMI, and STEMI groups had higher serum levels of LECT-2. In addition, the level of LECT-2 was related to the SYNTAX score and SYNTAX II score. Finally, patients with acute myocardial infarction (AMI) with elevated levels of LECT-2 had a higher risk of major adverse cardiovascular events (MACEs) within 12 months than those with lower levels of LECT-2. Plasma LECT-2 levels may be useful for the diagnosis of CAD and as predictors of MACE in patients with AMI.

Novel leukocyte-depleted platelet-rich plasma-based skin equivalent as an in vitro model of chronic wounds: a preliminary study

BackgroundChronic leg ulcerations are associated with Haemoglobin disorders, Type2 Diabetes Mellitus, and long-term venous insufficiency, where poor perfusion and altered metabolism develop into a chronic inflammation that impairs wound closure. Skin equivalent organotypic cultures can be engineered in vitro to study skin biology and wound closure by modelling the specific cellular components of the skin. This study aimed to develop a novel bioactive platelet-rich plasma (PRP) leukocyte depleted scaffold to facilitate the study of common clinical skin wounds in patients with poor chronic skin perfusion and low leukocyte infiltration. A scratch assay was performed on the skin model to mimic two skin wound conditions, an untreated condition and a condition treated with recombinant tumour necrotic factor (rTNF) to imitate the stimulation of an inflammatory state. Gene expression of IL8 and TGFA was analysed in both conditions. Statistical analysis was done through ANOVA and paired student t-test. P < 0.05 was considered significant.ResultsA skin model that consisted of a leukocyte-depleted, platelet-rich plasma scaffold was setup with embedded fibroblasts as dermal equivalents and seeded keratinocytes as multi-layered epidermis. Gene expression levels of IL8 and TGFA were significantly different between the control and scratched conditions (p < 0.001 and p < 0.01 respectively), as well as between the control and treated conditions (p < 0.01 and p < 0.001 respectively). The scratch assay induced IL8 upregulation after 3 h (p < 0.05) which continued to increase up to day 1 (p < 0.05). On the other hand, the administration of TNF led to the downregulation of IL8 (p < 0.01), followed by an upregulation on day 2. IL8 gene expression decreased in the scratched condition after day 1 as the natural healing process took place and was lower than in the treated condition on day 8 (p < 0.05).Both untreated and treated conditions showed a downregulation of TGFA 3 h after scratch when compared with the control condition (p < 0.01). Administration of rTNF showed significant downregulation of TGFA after 24 h when compared with the control (p < 0.01) and treated conditions (p < 0.05).ConclusionThis study suggests that a leukocyte-depleted PRP-based skin equivalent can be a useful model for the in vitro study of chronic skin wounds related to poor skin perfusion.

Sulfatase 2 Is Associated with Steroid Resistance in Childhood Nephrotic Syndrome.

Glucocorticoid (GC) resistance complicates the treatment of ~10–20% of children with nephrotic syndrome (NS), yet the molecular basis for resistance remains unclear. We used RNAseq analysis and in silico algorithm-based approaches on peripheral blood leukocytes from 12 children both at initial NS presentation and after ~7 weeks of GC therapy to identify a 12-gene panel able to differentiate steroid resistant NS (SRNS) from steroid-sensitive NS (SSNS). Among this panel, subsequent validation and analyses of one biologically relevant candidate, sulfatase 2 (SULF2), in up to a total of 66 children, revealed that both SULF2 leukocyte expression and plasma arylsulfatase activity Post/Pre therapy ratios were greater in SSNS vs. SRNS. However, neither plasma SULF2 endosulfatase activity (measured by VEGF binding activity) nor plasma VEGF levels, distinguished SSNS from SRNS, despite VEGF’s reported role as a downstream mediator of SULF2’s effects in glomeruli. Experimental studies of NS-related injury in both rat glomeruli and cultured podocytes also revealed decreased SULF2 expression, which were partially reversible by GC treatment of podocytes. These findings together suggest that SULF2 levels and activity are associated with GC resistance in NS, and that SULF2 may play a protective role in NS via the modulation of downstream mediators distinct from VEGF.

The Effect of Platelet-Rich Plasma Leukocyte Concentration on Arthroscopic Rotator Cuff Repair: A Network Meta-analysis of Randomized Controlled Trials

Background: It is unclear whether leukocyte-poor (LP) or leukocyte-rich (LR) varieties of platelet-rich plasma (PRP) as an adjuvant to arthroscopic rotator cuff repair (ARCR) result in improved tendon healing rates. Purpose: To perform a network meta-analysis of the randomized controlled trials in the literature to ascertain whether there is evidence to support the use of LP- or LR-PRP as an adjunct to ARCR. Methods: The literature search was based on the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. Randomized controlled trials comparing LP- or LR-PRP with a control alongside ARCR were included. Clinical outcomes, including retears and functional outcomes, were compared using a frequentist approach to network meta-analysis, with statistical analysis performed using R. The treatment options were ranked using the P-score. Results: There were 13 studies (868 patients) included, with 9 studies comparing LP-PRP with a control and 4 studies comparing LR-PRP with a control. LP-PRP was found to significantly reduce the rate of retear and/or incomplete tendon healing after fixation, even among medium-large tears; it also improved outcomes on the visual analog scale for pain, Constant score, and University of California Los Angeles score. LP-PRP had the highest P-score for all treatment groups. LR-PRP did not result in any significant improvements over the control group, except for visual analog scale score for pain. However, post hoc analysis revealed that LP-PRP did not lead to significant improvements over LR-PRP in any category. Conclusion: The current study demonstrates that LP-PRP reduces the rate of retear and/or incomplete tendon healing after ARCR and improves patient-reported outcomes as compared with a control. However, it is still unclear whether LP-PRP improves the tendon healing rate when compared with LR-PRP.

B cell-specific GPR55 deficiency promotes atherosclerosis

Abstract Background Atherosclerosis is accompanied by an imbalance between resolving and pro-inflammatory lipid mediators. Targeting lipid signaling pathways might offer a new anti-inflammatory therapy for improving the clinical outcome in cardiovascular disease patients. We considered lysophosphatidylinositol (LPI) and its receptor G protein-coupled receptor (GPR)55 as a potential modulator of atherosclerosis. Its role in regulating atherosclerosis and B cell function is unknown. Hypothesis We assessed the hypothesis that GPR55 signaling causally affects atherosclerosis and whether it has a specific role in regulating B cell function in this disease. Methods Atherosclerotic plaques were compared between apolipoprotein E deficient (ApoE−/−) and ApoE−/−Gpr55−/− mice after 4 to 16 weeks Western Diet (WD; 0.15% cholesterol; n=12–15 per group). To specifically test the role of B cell GPR55 in atherosclerosis, we generated mixed chimeras by lethally irradiating low density lipoprotein receptor deficient (Ldlr−/−) mice and reconstituting with a mixture of μMT and wildtype (control) or μMT and Gpr55−/− bone marrow cells. Circulating B cells were sorted and bulk RNA sequencing analysis was performed. We performed lipid and immunostainings of murine aortic root plaques, qPCR and ELISA of tissue lysates, as well as multiplex analysis of plasma immunoglobulins. Leukocyte plasma and tissue counts were determined by flow cytometry. Results GPR55 expression in mouse and human atherosclerotic plaques was detected by immunostaining. Furthermore, we confirmed murine Gpr55 mRNA expression on sorted circulating B220+B cells via qPCR, which was higher compared to CD3+ T cells, while CD11+ myeloid cells as well as NK cells had only low Gpr55 mRNA levels. ApoE−/−Gpr55−/− mice had significantly larger plaques after 4&amp;16 weeks WD compared to ApoE−/− controls, with more pronounced body weight increases and higher cholesterol levels at the 16 weeks WD time point. In addition, global Gpr55 deficiency resulted in enhanced aortic pro-inflammatory cytokine mRNA expression (IL-1β, IL-6, TNFα) and a massively upregulated IgG1 plasma levels and increased percentages of splenic germinal center and plasma cells. B-cell RNA-seq analysis showed 460 differential expressed regulated genes in the ApoE−/−Gpr55−/− compared to ApoE−/−. The main pathways affected were calcium ion transport, immunoglobulin production, negative regulation of phosphorylation, and cellular component morphogenesis, suggesting a dsysregulation of B cell function. B cell specific Gpr55 deficiency blunted the metabolic effects on body weight and cholesterol, but still translated in larger atherosclerotic plaques and elevated plasma IgG levels compared to the respective controls. Conclusion Both global and B cell-restricted Gpr55 deficiency promotes atherosclerosis and is associated with a more pro-inflammatory phenotype. Our findings suggest a novel role for GPR55 in regulating B cell development and function. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Deutsche Forschungsgemeinschaft (DFG)

Inter-individual differences in the response to an exercise training intervention in patients with metabolic syndrome

Abstract Background Regular physical exercise and standardized exercise training (ET) programmes positively affect a number of metabolic and cardiovascular risk factors. Yet, response to ET varies between patients and between individual risk parameters. We investigated inter-individual responses regarding blood pressure, heart rate, lipids and glucose metabolism, as well as physical fitness to a 16-weeks supervised exercise training protocol in patients with metabolic syndrome (MetS). Methods 29 MetS patients (20M/9F, age 52–70 y, BMI 24.9–38.2 kg/m2) exercised for 16 weeks in a supervised setting with varying volume and intensity. At baseline and at completion of the programme, blood pressure, heart rate, plasma lipids, glucose and insulin levels, as well as physical fitness were assessed, as were quantitative and morphological leukocyte parameters, cytokines and 86 metabolically relevant plasma proteins. Associations between individual risk parameters, leukocyte profile and plasma proteins were explored by recursive partitioning and network analysis. T-distributed stochastic neighbor embedding and hierarchical clustering were used for clustering based on cardiometabolic target parameters at 16 weeks. Results In the whole cohort, VO2peak increased and plasma counts of total leukocytes, neutrophils, monocytes (classical and intermediate), NK cells and effector and regulatory CD4+ lymphocytes, as well as circulating endothelial microvesicles were reduced at the end of the ET programme versus baseline. Plasma levels of carboxylesterase 1 decreased and soluble interleukin-7 receptor increased. Based on patient's cardio-metabolic parameters, three clusters were identified: cluster 1 had lower values of triglycerides, total and LDL cholesterol, cluster 2 had higher levels of triglycerides and cluster 3 had a greater heart rate recovery and lower insulin levels, but higher total and LDL cholesterol levels at follow-up. In cluster 1, absolute and relative NK cell counts were decreased and no morphological activation was observed. In cluster 2, reduced relative counts of NK-T cells were decreased, but their size was increased. In cluster 3, size of most effector leukocyte subtypes (CD8+, CD4+ Teff, NK-T, neutrophils) was increased at follow-up versus baseline. Network analysis suggested four distinct networks of (1) triglycerides, (2) VO2peak, (3) diastolic BP and (4) a cluster containing total and LDL cholesterol, insulin and HR recovery, each associated with partly distinct sets of plasma proteins and leukocyte parameters. Conclusion Despite improved physical fitness in all patients participating in the 16 weeks ET programme, inter-individual differences regarding cardio-metabolic target parameters and leukocyte activation was evident. Identification of specific leukocyte responses and of plasma protein “fingerprints” might help to optimize patient-specific prevention programmes, integrating medication and lifestyle optimization. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): DZHK, DSHF

Investigation of seminal plasma chitotriosidase-1 and leukocyte elastase as potential markers for 'silent' inflammation of the reproductive tract of the infertile male - a pilot study.

Inflammatory mediators - chitotriosidase-1 (CHIT1) and leukocyte elastase (LE) - were analyzed in human seminal plasma in relation to total antioxidative status (TAS) and pro-inflammatory markers IL-1β and IL-6. Samples collected from 34 males who were part of infertile couples were divided into normozoospermic (N; n = 12, without symptoms of inflammation), oligozoospermic (O; n = 11) and teratozoospermic (T; n = 11) groups. significant differences were observed only in CHIT1 concentration between N and O samples. However, a higher mean LE concentration was also observed in O and T patients (3.7-times and 900-times, respectively) compared with the N group. in IL-1β and IL-6 concentrations, an upward trend was observed from N, through O, up to the T group. The positive correlation between the concentration of IL-1β and the activity and specific activity of CHIT11 as well as the moderate negative correlation between concentrations of IL-1β and CHIT1 may suggest that elevated CHIT11 levels appeared in early stages of inflammation before the increase in IL-1β concentrations, or remained stable even after the levels of cytokine decreased. The above seem to confirm the role of CHIT1 in the manifestation of 'silent' inflammation at a very early stage. To conclude, CHIT1 concentration appears to be an interesting biomarker that signals the presence of possible 'silent' inflammation accompanying oligozoospermia. We cannot draw such conclusions regarding LE concentration, because, although we observed differences in the mean values and medians between analyzed groups, they were not significant. The utility of CHIT1 in the follow-up of oligozoospermia-associated 'silent' subclinical inflammation is promising, but further studies on a larger patient test set are required.

Skeletal Muscle Interleukin-6 Contributes to the Innate Immune Response in Septic Mice.

Interleukin-6 (IL-6) is a major cytokine released by skeletal muscle. Although IL-6 plays complex but well-known roles in host defense, the specific contribution of skeletal muscle IL-6 to innate immunity remains unknown. We tested its functional relevance by exposing inducible skeletal muscle IL-6 knockdown (skmIL-6KD) mice to a cecal slurry model of polymicrobial peritonitis and compared responses to strain-matched controls and skeletal muscle Cre-matched controls at 3, 6, and 12 h postinfection. In both sexes, skmIL-6KD mice at 6 h of infection exhibited marked changes to leukocyte trafficking in the peritoneum, characterized by ∼1.75-fold elevation in %neutrophils, a ∼3-fold reduction in %lymphocytes and a ∼2 to 3-fold reduction in %basophils. A similar pattern was seen at 12 h. No changes were observed in plasma leukocyte counts. Circulating cytokines in female skmIL-6KD mice at 6 h consistently showed modest reductions in IL-6, but marked reductions in a broad range of both pro- and anti-inflammatory cytokines, e.g., TNFα and IL-10. In both sexes at 12 h, a generalized suppression of plasma cytokines was also seen after the effects of Cre-induction with raloxifene were addressed. There were no significant effects of skmIL-6KD on mortality in either sex. Collectively, our results are consistent with skmIL-6 playing an important and previously unrecognized role in immune cell trafficking and cytokine regulation during septic shock.

Membrane Compartmentalization and Scaffold Proteins in Leukocyte Migration.

Leukocyte migration across vessels into and within peripheral and lymphoid tissues is essential for host defense against invading pathogens. Leukocytes are specialized in sensing a variety of guidance cues and to integrate environmental stimuli to navigate in a timely and spatially controlled manner. These extracellular signals must be transmitted across the leukocyte’s plasma membrane in a way that intracellular signaling cascades enable directional cell movement. Therefore, the composition of the membrane in concert with proteins that influence the compartmentalization of the plasma membrane or contribute to delineate intracellular signaling molecules are key in controlling leukocyte navigation. This becomes evident by the fact that mislocalization of membrane proteins is known to deleteriously affect cellular functions that may cause diseases. In this review we summarize recent advances made in the understanding of how membrane cholesterol levels modulate chemokine receptor signaling and hence leukocyte trafficking. Moreover, we provide an overview on the role of membrane scaffold proteins, particularly tetraspanins, flotillins/reggies, and caveolins in controlling leukocyte migration both in vitro and in vivo.

Fator de condição relativo e hematologia de reprodutores de pacu Piaractus mesopotamicus (Holmberg, 1887)

Objetivou-se avaliar o fator relativo de condição relativo e os parâmetros hematológicos de machos e fêmeas de pacu Piaractus mesopotamicus após o processo de indução hormonal e desova. Foi realizada pesquisa de campo, com medidas quantitativas e qualitativas em que foram avaliados 30 peixes saudáveis, coletados aleatoriamente, sendo 14 fêmeas e 16 machos. Procedeu-se então uma coleta de sangue para análise dos seguintes parâmetros: hematócrito, proteína plasmática total, número de eritrócitos, hemoglobina, índices hematimétricos, volume corpuscular médio (VCM) e concentração de hemoglobina corpuscular média (CHCM). Contagem total e diferencial de leucócitos e trombócitos e a glicose plasmática também foram determinadas. O fator de condição foi semelhante para ambos os sexos, com valores próximos a 1. A proteína plasmática total e o número de neutrófilos circulantes foram maiores nas fêmeas (P&lt;0,05). Os demais parâmetros permaneceram inalterados. Essas diferenças constituem uma resposta de adaptação ao manejo. Todos os valores foram próximos aos relatados anteriormente para o pacu, P. mesopotamicus, em condições de higidez no sistema de cultivo intensivo, indicando que processo de indução e desova não causa prejuízos ao bem-estar dos peixes a ponto de desencadear alterações no fator de condição e parâmetros hematológicos.

ROLE OF ENDOGENIC INTOXICATION IN MUSCLE INJURY IN EXPERIMENT

Background. Endogenous intoxication is a multicomponent complex process due to the endogenous biological products or dysfunction of systemic natural detoxification. &#x0D; Objective. The aim of the research was to study the dynamics of indices of endogenous intoxication in rats with traumatic muscle damage in the experiment.&#x0D; Methods. The experiment was performed on 45 non-linear white rats, which were modeled with traumatic muscle damage. The level of endogenous intoxication was assessed by the content of medium plasma molecules (MMM), leukocyte and erythrocytic index of intoxication (LII and EII). The research was conducted on the 1st, 3rd, 7th, 14th days after the injury.&#x0D; Results. It was found that traumatic muscle damage causes endotoxemia. Manifestations of endogenous intoxication are: the increase of MMM1 in 2.3 times, MMM2 in 2.8 times compare to the intact animals. The level of this indicator slightly decreased in 7 days. Simultaneously with an increase in the MCT level in the post-traumatic period, the total toxic effect on the erythrocyte membrane also increased, which was manifested by a significant increase in EII in all terms of observation.&#x0D; Conclusions. Traumatic damage of the muscles is accompanied by the growth of molecules of average mass in upto 7 days of observation, which significantly differ from the indicators of the intact group. The results of our research prove that traumatic muscle damage causes endotoxicosis development evidenced by accumulation of endotoxins in the animals’ body that is proved by significant changes in endogenous intoxication indices: i.e. erythrocytic and leukocytic indexes of intoxication and content of medium mass molecules.

Surgical technique for persistent macular holes with application of the leucocyte and platelet rich plasma obtained with the Ycellbio system

Surgical technique for persistent macular holes with application of the leucocyte and platelet rich plasma obtained with the Ycellbio system

Plasma proteins and leukocyte kinetics of turtles (Podocnemis unifilis (Troschel, 1848)) inoculated with inactivated Escherichia coli

The plasma protein fractions and leukocyte response induced by an experimental subcutaneous inoculation of Escherichia coli isolated and inactivated in turtlesin captivity (Podocnemis unifilis—“tracaja”), were evaluated. The animals were divided into two groups (test group and control) with 10 turtles each. Blood samples from all animals were collected the day before the inoculation and at days 15, 28, 42 and 125 days in both groups. The inoculation of E. coli did not significantly alter the red blood cell count, but the serial collection of blood led to a mild normochromic macrocytic anemia.The inoculation of E. coli was not responsible for the observed changes in erythrogram. The leukocyte count showed an inflammatory process indistinct for the presence of E. coli, except for the lymphocytosis observed in the test group. The determination of the A:G ratio proved useful in the detection of immunoglobulin production responsive to inoculation with E. coli.