Increase In Number Of Platelets Research Articles

Testosterone replacement has a beneficial effect on the hemostatic system by altered gene expression of coagulation factors

This study aimed to investigate the effects of testosterone replacement therapy on hemostasis and some procoagulant gene expression in mice. 42 mice were randomly divided into two groups of non-orchiectomized (non-ORX) and orchiectomized (ORX) with three subgroups (n = 7) each, were subcutaneously administered with sesame oil (control), 2 and 20 mg/kg/week testosterone enanthate. Orchiectomized mice were allowed to recover for one week before treatment. On the 7th week of treatment, blood samples were collected for coagulation parameters analysis and measurement of plasma testosterone levels. Moreover, quantitative real-time PCR analysis was performed on liver samples to assess the expression of factor IX, factor X, and prothrombin genes. The results showed that supraphysiological doses (20 mg/kg) of testosterone significantly increased plasma testosterone levels in all groups, while physiological doses (2 mg/kg) only increased testosterone levels in non-ORX animals. Although testosterone administration had no effect on prothrombin time (PT) and activated partial thromboplastin time (aPTT), supraphysiological doses reduced bleeding time and clotting time. Furthermore, platelet count increased in a dose-dependent manner with testosterone enanthate treatment. The expression of coagulation factors was also decreased with supraphysiological doses of testosterone. In conclusion, testosterone had significant effects on primary hemostasis and common coagulation pathway, including increased platelet number and aggregation, decreased clotting time, and altered gene expression of coagulation factors.

Elevated Platelet Aggregation in Patients with Ovarian Cancer: More than Just Increased Platelet Count.

Background: Patients with ovarian cancer have high platelet counts, which correlate with disease burden, incidence, and lethality of blood clots (thrombosis). We hypothesized that elevated aggregation is associated with both increased platelet number and altered behavior of platelets in patients with ovarian cancer. Methods: Healthy controls and patients with suspected or diagnosed ovarian cancer were evaluated for complete blood counts. To evaluate the effects of platelet count versus platelet behavior, equal platelet-rich plasma (PRP) volumes versus equal platelet numbers were used in platelet aggregation assays. Arachidonic acid, adenosine diphosphate, and collagen platelet agonists were used to induce aggregation. Volunteers were grouped into healthy controls (23), benign/borderline cases (7), and cancer cases (25 ovarian, 1 colorectal, and 2 endometrial). Results: The rate and amount of platelet aggregation were higher in patients compared to healthy controls regardless of whether the same platelet number or PRP volume was used. Compared to healthy controls, patients with untreated ovarian cancer exhibited high levels of platelet activation markers, P-selectin (27.06 vs. 31.06 ng/mL, p = 0.03), and beta-thromboglobulin (3.073 vs. 4.091 µg/mL, p = 0.02) in their plasma. The significance of the elevation and its correlations with platelet number or PRP volume varied depending on the agonist. Platelet (305.88 vs. 134.12, p < 0.0001) and white blood cell (8.459 vs. 5.395, p < 0.01) counts (×109/L) were elevated pre-chemotherapy and decreased post-chemotherapy, respectively. Conclusions: Elevated platelet aggregation is caused by both altered platelet number and behavior in patients with ovarian cancer. These results support the study of antiplatelet agents for thrombosis prevention in these patients.

From in silico to in Vitro to In Vivo: Increasing Megakaryocytes to Treat Thrombocytopenia

Hematopoietic stem cells undergo a multi-step differentiation program through several precursor stages to give rise to megakaryocytes (Mk), which ultimately release platelets. Thrombocytopenia can manifest in various clinical settings and can adversely affect a patient's quality of life through easy bruising and risk of bleeding. An example is chemotherapy-induced thrombocytopenia (CIT) which occurs when myelosuppressive chemotherapy is given, especially with certain agents such as platinum analogs and cyclophosphamide. We used a single-cell RNA-sequencing approach to study the differentiation of hematopoietic stem cells to Mk progenitors (MkP) and delineated the trajectory that defines the transition of cells to the megakaryocytic lineage. Using our proprietary platform which pairs new approaches in computation and machine learning in drug discovery, we predicted small molecule interventions that could modulate the hematopoietic lineage direction to increase the differentiation towards MkP and subsequent increases in platelet counts. Using an in vitro 5-day CD34-based Mk differentiation assay we demonstrated that some of our predicted interventions increased generation of MkP from stem cells. Analysis of early MkP (CD34-CD235a-CD71+CD41b+CD42b-) and late MkP (CD34-CD235a-CD71-CD41b+CD42b+) by flow cytometry elucidated differential mechanisms of action such as induction of early or late MKP or both. We performed an 8-day study in healthy mice, which revealed that a selected set of compounds increased MkPs in vivo, consistent with our predicted cell behavior and in vitro findings. To evaluate a possible application for this MkP and platelet increases we tested the efficacy of one of our hit compounds CP3 in a CIT mouse model using Carboplatin. In this model, we recorded a &amp;gt;70 % reduction in platelet counts on day 9 post carboplatin administration, with platelet numbers recovering on day 13 and return to basal levels by day 21. To assess the efficacy of CP3, mice dosed first with this compound were challenged with carboplatin. The primary endpoint of this study was the ability of the compound to rescue mice from nadir of thrombocytopenia and accelerate recovery to baseline. Prior to carboplatin dosing, 7 days of CP3 dosing showed a dose dependent increase in platelet counts confirming our initial studies. However, CP3 dosing did not rescue platelet nadir or accelerate recovery to baseline in the CIT model. This study demonstrated the capacity of CP3 to increase platelets in a healthy BM environment but not under conditions of myelosuppression as seen in the CIT mouse model. Taken together, these findings highlight the benefits of our platform to predict compounds that modulate lineage differentiation towards MkP, and a resultant increase in platelet numbers in an animal model.

Thrombotic thrombocytopenic purpura in a patient on long-term alpha-interferon therapy for essential thrombocythemia: a case report

BackgroundThrombotic thrombocytopenic purpura (TTP) is rare and severe thrombotic microangiopathy characterized by thrombocytopenia, hemolytic anemia, and renal dysfunction. In contrast, essential thrombocythemia (ET) is a myeloproliferative disease associated with an abnormal increase in platelet numbers. Previous studies reported several cases of the development of ET in patients with TTP. However, the case of an ET patient complicated with TTP has not been previously reported. In this case study, we present a patient with TTP who was previously diagnosed with ET. Therefore, to the best of our knowledge, this is the first report of TTP in ET.Case presentationA 31-year-old Chinese female who was previously diagnosed with ET presented with anemia and renal dysfunction. The patient had been on long-term treatment with hydroxyurea, aspirin, and alpha interferon (INF-α) for ten years. The diagnosis of TTP was confirmed by clinical features, schistocytes noted on the peripheral blood smear, and lower ADAMTS13 activity (8.5%), together with the renal biopsy results. INF-α was discontinued, and the patient was then treated with plasma exchange and corticosteroids. After one year of follow-up, the patient had a normal hemoglobin level and platelet numbers, and her ADAMTS13 activity had improved. However, the patient’s renal function remains impaired.ConclusionsWe report a case of an ET patient complicated with TTP that was possibly due to INF-α, highlighting the potential complications associated with long-term ET therapy. The case also highlights the importance of considering TTP in patients with pre-existing ET who present with anemia and renal dysfunction, extending the spectrum of known studies.

Platelet Indices: Is it a Reliable Biomarker in Viral Infections?

Platelets are non-nucleated blood cells that are essential for maintaining hemostasis. While platelet activation and increased platelet numbers (thrombocytosis) are associated with a higher risk of thrombotic events, low platelet counts (thrombocytopenia) and a variety of platelet function abnormalities are associated with a higher risk of bleeding. Platelets and their activation state have been proven in recent years to influence innate and adaptive immune responses, and low platelet counts have been identified as a surrogate indicator for poor prognosis in septic patients. Platelet activation frequently occurs in conjunction with viral infections. Viruses, on the other hand, can interact directly with platelets and megakaryocytes, modulating their function. All these factors contribute to platelet activation, which leads to increased platelet consumption and elimination, as well as thrombocytopenia, which is common during viral infection Platelets and their secreted products have been demonstrated to suppress infection and aid virus persistence in the presence of certain viruses, making platelets a double-edged sword during viral infections. In this review we have highlighted about what is currently known about platelet interactions with various types of viruses, virus effects on platelet activation, as well as platelet-mediated regulation of innate and adaptive immune responses.

The role of tumor-educated platelets in ovarian cancer: A comprehensive review and update.

Platelets have recently surfaced as critical players in cancer metastasis and the local and systemic responses to tumor growth. The emerging concept of "Tumor-educated platelets (TEPs)" comprises the exchange of biomolecules between tumor cells and platelets, thereby leading to the "education" of platelets. Increased platelet numbers have long been associated with cancer patients' tumor metastasis and poor clinical prognosis. However, it is very recently that researchers have delved deeper into the tumor-microenvironment and probed the mechanism of interactions between tumor cells and platelets. Designing strategies to target the TEPs and the communications between platelets and tumor cells can prove to be a promising breakthrough in cancer therapy. Through this review, we aim to analyze the recent developments in this field and discuss the characteristics of TEPs, focusing on ovarian cancer-associated TEPs and their characteristics, the interplay between ovarian cancer-associated TEPs and cancer cells, and the purview of TEP-targeted cancer diagnosis and therapy, including platelet biomarkers and inhibitors.

Impact of hypovitaminosis D on body weight, food intake parameters and hematological alterations in albino mice Mus musculus

Hypovitaminosis D is a severe global public health issue. Vitamin D is a fat-soluble secosteroid hormone that may be obtained by diet as well as sun exposure. The efficacy of the vitamin D receptor and vitamin D activating enzyme present in tissues and cells, along with the influence of vitamin D on the musculoskeletal system, support vitamin D's extensive functions. Insufficient vitamin D intake, results in hepatic, renal, skin, and hematological problems which signify vitamin D deficiency. A heat-dependent phase in the synthesis of vitamin D transforms 7-dehydrocholesterol into previtamin D 3 , which is subsequently isomerized into cholecalciferol. The present investigation demonstrates the effect of a long-term inadequate diet on body weight, food intake, and hematological parameters. In this study, female Swiss albino mice were divided into three groups: the control group received standard feed, the vitamin-D-supplemented group received cholecalciferol (1000IU/kg), and the vitamin-D deficient group was fed with a diet lacking in vitamin D for 90 days. Vitamin-D deficient animal group had higher levels of parathyroid hormone and lower levels of 25 hydroxy vitamin D [25(OH)D] in their serum as compared to control animals (p < 0.0001). A substantial decrease in hemoglobin, red blood cells, white blood cells and hematocrit values was observed in the vitamin-D deficient group, along with a significant increase in platelet number. Our findings conclude that consuming a long-term vitamin D-deficient diet changes hematological, dietary, and body weight factors and may result in severe in long-term health problems. • Hypovitaminosis D is inversely proportional with PTH serum level in long term consumption of vitamin D deficient diet. • Daily food consumption of a vitamin D deficient diet resulted in noticeably changed hematological indicators and minor alterations in weight factors. • Vitamin D supplementation in regular basis improve in 25(OH)D serum level and moderately balance PTH. • Higher PTH level and hypocalcemia is directly associated with renal hydroxylation in vitamin D metabolism.

Toxicological Studies of Hydroethanolic Leaf Extract of Xylopia aethiopica (Dunal) A. Rich. (Annonaceae) on Wistar Rats

Introduction: The impact of aromatic plants used in cooking on health is indisputable.&#x0D; This study aims to evaluate the in vitro cytotoxicity of Artemia salina, acute and the 28 days subchronic toxicity of X. aethiopica dried fruit used like spice in Togo by oral administration of the extract to female Wistar rats.&#x0D; Results: The A. salina assay showed that the hydroalcoholic extract of X. aethiopica presented any toxicity. The LC50 values of X. aethiopica on brine shrimp were 0.64 ± 0.13 mg/mL. The acute toxicity study revealed neither behavioral disturbances nor death in rats. The lethal dose (DL50) of this extract is greater than 5000 mg/kg body weight. The results of subchronic toxicity showed no significant change in body and organs weight gain in rats at test doses throughout the treatment period. No changes in haematological parameters were observed apart from a significant increase in platelet number at doses of 500 and 1000 mg/kg. Biochemical parameters such as Urea, Creatinine, Triglycerides, Total cholesterol, Serum glutamic pyruvic transaminase (SGPT), C-reactive protein (CRP) and glycemia were not significantly changed. Serum glutamic oxaloacetic transaminase (SGOT) and Alkaline phosphatase (ALP) activities increased at doses of 500/1000 mg/kg and 1000 mg/kg respectively. Creatine Phosphokinase (CPK) activity decrease at the dose of 500 and 1000 mg/kg. Blood electrolytes showed no significant change. The histological sections showed no organ damage.&#x0D; Conclusions: Aqueous extract of X. aethiopica did not lead to any adverse effects in rats after acute and subchronic treatment at 500 and 1000 mg/kg doses. &#x0D; Keywords: Xylopia aethiopica, Cytotoxicity, Toxicity, Wistar rats.

Effect of vitamin D on myocardial remodeling and inflammatory status in children with congenital heart disease.

Vitamin D insufficiency is a common public health problem that is often unrecognized in children with congenital heart disease, and is not generally evaluated at congenital heart disease (CHD) follow-up. Recent studies have suggested that inadequate vitamin D status may have an adverse effect on cardiovascular health. This study investigates the relationship between vitamin D levels and hemodynamic parameters in children with CHD. Included in the study 40 patients (25 females, 15 males) with CHD, who were evaluated for Ross heart failure score, vitamin D, parathyroid hormone (PTH), calcium, phosphorus, alkaline phosphatase (ALP), whole blood count (WBC) and echocardiographic measurements, and all measurements were repeated in the third month of the therapy. The mean vitamin D level was 16.4 ± 6.6 ug/L before and 27.5 ± 9.9 μg/L in the third month of therapy, while the mean PTH level was 53.3 ± 34.9 pg/mL before and 43.8 ± 21.4 pg/mL in the third month of therapy. The mean WBC was 8084 ± 2324/µL before and 7378±1893/µL in the third month of the therapy, and the mean platelet (PLT) count was 280,897 ± 80,119/µL before and 307,179 ± 60,202/µL in the third month of the therapy. The mean ejection fraction (EF) was 64% ± 7.2% before and 66.7% ± 6.2% in the third month of therapy, while the right ventricle (RV) myocardial performance index (MPI) was 32.1% ± 6.7% before and 28.9% ± 6.5% in the third month of the therapy. IL10 level was increased in four patients in the third month of therapy. A statistically significant decrease in PTH level and WBC, and an increase in PLT number and IL-10 level were detected by the therapy. Furthermore, echocardiographic findings revealed a statistically significant increase in EF and a decrease in RVMPI attributable to the therapy. The decreased levels of PTH, which is a proinflammatory marker, the increases in IL-10, which is an antiinflammatory cytokine, and the decreases in the number of WBC resulting from vitamin D treatment demonstrate the antiinflammatory effects of vitamin D. An improvement in EF means improvement in left ventricular contractility, while a decrease in RV MPI has been shown to improve the systolic and diastolic function of the right ventricle. These results suggest that vitamin D therapy has a positive effect on the heart, and so vitamin D levels should be evaluated during the routine follow-up of congenital heart disease.

Platelet CLEC2-Podoplanin Axis as a Promising Target for Oral Cancer Treatment.

Cancer tissues are not just simple masses of malignant cells, but rather complex and heterogeneous collections of cellular and even non-cellular components, such as endothelial cells, stromal cells, immune cells, and collagens, referred to as tumor microenvironment (TME). These multiple players in the TME develop dynamic interactions with each other, which determines the characteristics of the tumor. Platelets are the smallest cells in the bloodstream and primarily regulate blood coagulation and hemostasis. Notably, cancer patients often show thrombocytosis, a status of an increased platelet number in the bloodstream, as well as the platelet infiltration into the tumor stroma, which contributes to cancer promotion and progression. Thus, platelets function as one of the important stromal components in the TME, emerging as a promising chemotherapeutic target. However, the use of traditional antiplatelet agents, such as aspirin, has limitations mainly due to increased bleeding complications. This requires to implement new strategies to target platelets for anti-cancer effects. In oral squamous cell carcinoma (OSCC) patients, both high platelet counts and low tumor-stromal ratio (high stroma) are strongly correlated with increased metastasis and poor prognosis. OSCC tends to invade adjacent tissues and bones and spread to the lymph nodes for distant metastasis, which is a huge hurdle for OSCC treatment in spite of relatively easy access for visual examination of precancerous lesions in the oral cavity. Therefore, locoregional control of the primary tumor is crucial for OSCC treatment. Similar to thrombocytosis, higher expression of podoplanin (PDPN) has been suggested as a predictive marker for higher frequency of lymph node metastasis of OSCC. Cumulative evidence supports that platelets can directly interact with PDPN-expressing cancer cells via C-type lectin-like receptor 2 (CLEC2), contributing to cancer cell invasion and metastasis. Thus, the platelet CLEC2-PDPN axis could be a pinpoint target to inhibit interaction between platelets and OSCC, avoiding undesirable side effects. Here, we will review the role of platelets in cancer, particularly focusing on CLEC2-PDPN interaction, and will assess their potentials as therapeutic targets for OSCC treatment.

Pre-Clinical Modeling of Concomitant Therapy with Azacytidine and Eltrombopag in MDS/AML

Hemorrhage-related morbidity and mortality frequently arises in patients with myelodysplastic syndromes (MDS). A major therapeutic goal in patients with MDS is the alleviation commonly occurring thrombocytopenia. Low platelet counts increase bleeding risk, limit tolerated doses and efficacy of standard therapies, and are associated with an increased risk of leukemic transformation. Preclinical and phase I/II studies on eltrombopag (EPAG), a megakaryopoiesis-stimulating thrombopoietin receptor (TPO-R) agonist, demonstrated safety and efficacy of the small molecule as monotherapy in MDS. However, a phase III study testing the ability of EPAG to increase platelet numbers in patients with intermediate or higher-risk MDS under azacytidine (AZA) therapy (NCT02158936) was prematurely terminated due to inferior performance of EPAG-treated vs. placebo-receiving patients. While the study did not find a difference in overall survival rates between the two groups, concerns pertaining to disease progression under concomitant treatment with EPAG and azacytidine have been raised.We assessed whether EPAG exerts antagonistic physiological activities in the context of anti-leukemia treatment with AZA exploiting a set of complementary pre-clinical ex vivo assays measuring aberrant cell proliferation, differentiation and self-renewal. MDS and AML patient-derived cell lines (n=4) and primary cell specimen (n=6) were treated with non-cytotoxic, clinically-relevant doses of EPAG and AZA in short-term liquid cultures, myeloid colony-forming assays, and long-term serial-replating assays. To elucidate functional effects of the combination treatment, we determined cell growth, apoptosis, differentiation and long-term colony initiation as a surrogate for self-renewal upon treatment. We also quantified cell surface presence of TPO-R, and assessed pathway activation downstream of the receptor using phosphoflow.Consistent with previous observations, continuous single agent treatment with either >0.3μM AZA, or 3-20mg/ml EPAG were found to be cytotoxic for leukemic cells. Simultaneous treatment with a non-cytotoxic functionally-relevant dosage of AZA (0.3μM) and EPAG at concentrations above 3mg/ml resulted in a striking decrease of cell viability compared to AZA single agent treatment in short-term cell line cultures. Lower-dose EPAG treatment (0.3 and 0.5mg/ml) along with AZA (0.3μM) did not lead to any significant changes in cell viability and growth in short term cultures (vs. single agent treated controls) of cell lines or primary cells. Apoptosis, cell surface differentiation marker analysis, and morphological assessment revealed no discernable differences upon combination treatment compared to AZA single agent treatment in cell lines. Characterization of aberrant self-renewal using serial replating colony assays revealed a significant decrease in the number of colony-initiating cells upon continuous single agent treatment with AZA, which was not diminished by combination treatment with EPAG. An AZA-resistant cell line also did not show altered long-term colony-forming cell numbers in the presence of both drugs. We found no evidence of increased leukemic self-renewal upon simultaneous treatment using primary cells in 5/6 specimen tested. One sample showed a transient increase in colony formation and average cell numbers per colony in the presence of both drugs, compared to AZA-treated controls, albeit the number of serial replatings was unchanged compared to AZA controls. We tested whether TPO-R/CD110-expressing aberrant cells would expand under EPAG single agent, or combination treatment with AZA, and found no significant changes in the frequency of CD110-positive cells compared to AZA single agent controls. TPO-R downstream signaling in primary mononuclear cells comprised a marked, yet transient impairment in STAT5 and AKT phosphorylation upon combination treatment, compared to single EPAG treatment.Togehter, our data show that simultaneous treatment of leukemic cells with non-cytotoxic, but clinically-relevant dosages of EPAG and AZA does not lead to a stimulation of leukemic cell growth, even in the presence of TPO-R, in several pre-clinical models. These results do not support a generalized mechanistic antagonism between the two drugs in leukemic cells. Further investigations are underway to understand the impact on non-leukemic hematopoiesis. DisclosuresSteidl:Bayer Healthcare: Consultancy; Novartis: Research Funding; Celgene: Consultancy; GlaxoSmithKline: Research Funding; Aileron Therapeutics: Consultancy, Research Funding. Marques Ramos:Novartis Pharmaceuticals: Employment. Pallaud:Novartis Pharmaceuticals: Employment. Will:Novartis Pharmaceuticals: Consultancy, Research Funding.

Platelets inhibit erythrocyte invasion by Plasmodium falciparum at physiological platelet:erythrocyte ratios.

To evaluate the effect of platelet:erythrocyte (P:E) ratios on Plasmodium falciparum erythrocyte invasion. Recent reports have shown that platelets are directly involved in the immune response towards P. falciparum during erythrocyte invasion. However, the literature both supports and conflicts with a role for platelets in limiting invasion. Also, the effect of platelet numbers on invasion (parasitemia) has not been thoroughly investigated. The P. falciparum strains FCR3S1.2 and W2mef were cultured with group O erythrocytes. The cultures were synchronised and supplemented with pooled platelets at P:E ratios ranging from 1:100 to 1:2. Parasitemia was measured at 40 h by flow cytometry and by microscopy of blood smears. A linear relationship was observed between reduced invasion and increased platelet numbers at P:E ratios ranging from 1:100 to 1:20. However, this effect was reversed at lower ratios (1:10-1:2). Microscopic evaluation revealed aggregation and attachment of platelets to erythrocytes, but not specifically to parasitised erythrocytes. We have shown that under physiological P:E ratios (approx. 1:10-1:40), platelets inhibited P. falciparum invasion in a dose-dependent manner. At ratios of 1:10 and below, platelets did not further increase the inhibitory effect and, although the trend was reversed, inhibition was still maintained.

Broad-Spectrum Antibiotics Deplete Bone Marrow Regulatory T Cells.

Bone marrow suppression, including neutropenia, is a major adverse effect of prolonged antibiotic use that impairs the clinical care and outcomes of patients with serious infections. The mechanisms underlying antibiotic-mediated bone marrow suppression remain poorly understood, with initial evidence indicating that depletion of the intestinal microbiota is an important factor. Based on our earlier studies of blood and bone marrow changes in a mouse model of prolonged antibiotic administration, we studied whether changes in megakaryocytes or regulatory T cells (Tregs), two cell types that are critical in the maintenance of hematopoietic stem cells, contribute to antibiotic-mediated bone marrow suppression. Despite increased platelet numbers, megakaryocytes were unchanged in the bone marrow of antibiotic-treated mice; however, Tregs were found to be significantly depleted. Exogenous addition of Tregs was insufficient to rescue the function of bone marrow from antibiotic-treated mice in both colony formation and transplantation assays. These findings indicate that the intestinal microbiota support normal Treg development to protect healthy hematopoiesis, but that the restoration of Tregs alone is insufficient to restore normal bone marrow function.

Sirolimus versus cyclosporine for the treatment of pediatric chronic immune thrombocytopenia: A randomized blinded trial

IntroductionChronic immune thrombocytopenia (ITP) of childhood is still a problem. For treating ITP, several immunosuppressive medications can be considered with various response rates. Our goal was to compare effects of sirolimus and cyclosporine on children with chronic ITP. MethodsThis randomized and blinded trial was carried out on 67 children over 5 years old with chronic ITP. Patients were assigned 1:1 to cyclosporine and sirolimus for 6 months. Platelet count was assessed and compared between 2 study groups at different intervals. The clinical trial registry number was IRCT20180501039499N1. ResultsSixty-one children completed the 6-month treatment. Mean age was 9.3 years with an excess of females. Compared to baseline values, both drugs caused a significant increase in number of platelets over the course of treatment; sirolimus group: 15,800/mcL vs 96,566/mcL, (P < 0.001), cyclosporine group: 14,400/mcL vs 111,266/mcL, P < 0.001,). In addition, differences of platelet number were statistically significant at some treatment intervals (3rd and 6th month, P < 0.05). A quicker response was observed in children receiving cyclosporine. Both drugs had similar rate of response which occurred in 50% of included patients. Finally, sirolimus had a better safety profile. ConclusionsOur study showed that cyclosporine and sirolimus had an equal rate of response in treating chronic ITP of children. At the same time, the two medications showed significant differences in their side effects.

Reactive thrombocytosis in malignancy- Can it be a strong predictor for IT

An increased platelet number may be secondary to many conditions. Malignancies are known to induce thrombocytosis in some cases. In patients with malignancies, thrombocytosis has previously been related to disease stage, histological type, and survival. Studies have shown that thrombocytosis is associated with a poor prognosis in various malignancies such as carcinoma ovary, cervical cancer, endometrial cancer, breast cancer and lung cancer. The aim of this study was to analyze the etiology and prevalence of thrombocytosis in malignancy and to assess whether platelet count can be used as a predictor of malignancy in the cases diagnosed as cancer at the time of its first diagnosis. This descriptive study was done on 500 patients with platelet count &amp;#62; 450,000/ μl with the cause being termed reactive. The most common cause of reactive thrombocytosis was Infections (28.8%), Tissue damage (16.4%), Iron deficiency anemia (16.2%), Malignancy (9.6%) and Inflammation (9.4%). Among malignancies, carcinoma oral cavity (20.8%) was found to be more commonly associated with thrombocytosis; with only (4.2%) cases being less than 18 years, rest (95.8%) cases were above 18 years. Thrombocytosis is associated with various neoplasms, therefore it can be used as a diagnostic clue for malignancy in an undiagnosed patient presenting with reactive thrombocytosis and associated symptoms of the disease thus indicating poor outcomes and mortality.

Effect of yeast culture (Saccharomyces cerevisiae) supplementation on growth performance, blood metabolites, carcass traits, quality, and sensorial traits of meat from pigs under heat stress

The objective of this study was to evaluate the effects of dietary supplementation of yeast culture (YC) in pigs under heat stress conditions on growth performance, blood metabolites, carcass traits, quality, and sensorial values of the meat. Thirty pigs (29.487 ± 1.46 kg) were stratified by body weight (BW) and randomly assigned to the following treatments: 1) Control group animals received only the basal diet; 2) 0.2 % YC group animals received the basal diet with the addition of 0.2 % YC; 3) 0.3 % YC group animals received the basic diet supplemented with 0.3 % YC. The study was conducted in three phases based on BW: phase I (30–65 kg), phase II (65–95 kg), and phase III (95–128 kg). In phase I, the supplementation with YC at 0.2 % and 0.3 % produced average daily gain (ADG) values that were higher by 25.52 % and 23.701 %, respectively (P < 0.05), average daily feed intake (ADFI) values that were higher by 13.42 % and 11.85 %, respectively (P < 0.05), and a higher final weight (by 8.09 % and 7.26 %; P < 0.05), compared with the control diet. In phase II, the final weight was 7.35 % higher with the addition of 0.3 % YC (P < 0.05) compared with the control. In phase III, the final weight was higher by 4.16 % and 4.67 % with the addition of YC at 0.2 % and 0.3 % (P < 0.05), respectively, compared with the control. During the whole period (29.487–128.701 kg), improvements in ADG were observed with both levels of YC supplementation (0.2 % and 0.3 % YC) by 5.38 % and 6.09 % (P < 0.05) compared with the control. In terms of blood metabolites, supplementation with 0.3 % YC increased platelet numbers compared with the addition of 0.2 % YC (P < 0.05). For carcass traits, a higher carcass weight was recorded with the YC treatments at 0.3 % and 0.2 % (by 4.63 % and 5.27 %, respectively: P < 0.05) compared with the control. In terms of meat quality, the use of 0.2 % YC caused a decrease in redness (a*) by 10.03 %, (P < 0.05) and in yellowness (b*) by 15.52 % in contrast with the control (P < 0.05). A trained panel detected minimal changes in the sensorial values of the meat with YC supplementation (P> 0.05). In conclusion, YC improved both growth and carcass weight without markedly changing blood metabolites, carcass traits, quality, and sensorial values of the meat from pigs under heat stress conditions.

Hydroxyvitamin D Serum Levels are Negatively Associated with Platelet Number in a Cohort of Subjects Affected by Overweight and Obesity.

Background: Hypovitaminosis D and higher platelet numbers are emerging as cardiovascular risk factors, in particular in obese subjects. Methods: This observational study was aimed at investigating the relationship between platelet number and serum 25-hydroxyvitamin D (25(OH)D) levels in a cohort of individuals affected by overweight and obesity (body mass index (BMI) ≥ 25 Kg/m2). A sample of 341 subjects (248 women, 93 men), aged 18–71 years, taking no medication, was examined. Anthropometric, hormone, metabolic and common routine hematochemical parameters were examined and evaluated in association with platelet count and serum 25(OH)D levels. Results: Platelet numbers were inversely related to age (p < 0.04), 25(OH)D (p < 0.05) and uric acid (p < 0.04) levels, and directly associated with white blood cells (p < 0.01), Thyroid Stimulating Hormone (TSH) (p < 0.04), insulin levels (p < 0.002) and Homeostasis Model Assessment – Insulin Resistance (HOMA-IR) (p < 0.002). We applied statistical regression models to examine the relationship between platelet count (dependent variable) and parameters that had univariate associations with platelet numbers, showing that the association between platelet count and 25(OH)D was not confirmed. Moreover, vitamin D showed a negative independent association with BMI, diastolic blood pressure and serum insulin levels. Conclusions: This study indicates, for the first time, that vitamin D deficiency is associated with a parallel increase in platelet number, suggesting that higher platelet numbers may be one of the possible mechanisms leading to a greater cardiovascular risk in obese subjects. It also shows that vitamin D deficiency, a common condition in obesity, has independent associations with higher BMI, diastolic blood pressure and serum insulin levels.

Phagocytosis by endothelial cells inhibits procoagulant activity of platelets of essential thrombocythemia in vitro

BackgroundEssential thrombocythemia (ET) is characterized by thrombocytosis with increased platelet number and persistent activation. The mechanisms of thrombosis and the fate of these platelets are not clear. The aim of the present study is to explore the phagocytosis of platelets of ET patients by endothelial cells (ECs) in vitro and its relevance to the procoagulant activity (PCA). MethodsPhosphatidylserine (PS) exposure on platelets was detected by flow cytometry. Phagocytosis of the platelets by ECs was performed using flow cytometry, confocal microscopy, and electron microscopy. The PCA of platelets was evaluated by coagulation time and purified coagulation complex assays. ResultsThe PS exposure on platelets in ET patients is higher than that in healthy controls. The PS‐exposed platelets are highly procoagulant and lactadherin reduced 80% of the PCA by blockade of PS. When cocultured, the platelets of ET patients were sequestered by ECs in a time‐dependent fashion. Lactadherin enhanced phagocytosis by bridging the PS on activated platelets and the integrin αvβ3 on ECs, and P‐selectin played at least a partial role in this process. Furthermore, factor Xa and prothrombinase activity of PS‐exposed platelets were decreased after incubation with ECs. ConclusionOur results suggest that phagocytic clearance of platelets by ECs occurs in ET patients, thus representing a novel mechanism to remove activated platelets from the circulation; lactadherin and phagocytosis could cooperatively limit the thrombophilia in ET patients.

A novel method for harvesting concentrated platelet-rich fibrin (C-PRF) with a 10-fold increase in platelet and leukocyte yields.

Liquid platelet rich fibrin (PRF; often referred to as injectable PRF) has been utilized as an injectable formulation of PRF that is capable of stimulating tissue regeneration. Our research group recently found that following standard L-PRF protocols (2700 RPM for 12 min), a massive increase in platelets and leukocytes was observed directly within the buffy-coat layer directly above the red blood cell layer. The purpose of this study was to develop a novel harvesting technique to isolate liquid PRF directly from this buffy coat layer and to compare this technique to standard i-PRF. Standard high g-force L-PRF and low g-force i-PRF protocols were utilized to separate blood layers. Above the red blood corpuscle layer, sequential 100-μL layers of plasma were harvested (12 layers total; i.e., 1.2 mL, which represents the total i-PRF volume), and 3 layers (3 × 100 μL) were harvested from the red blood cell layer to quantify blood cells. Each layer was then sent for complete blood count (CBC) analysis, and the cell numbers were quantified including red blood cells, leukocytes, neutrophils, lymphocytes, monocytes, and platelets. The liquid PRF that was directly collected from the buffy-coat layer following L-PRF protocols was referred to as concentrated PRF (C-PRF). The i-PRF protocol typically yielded a 2- to 3-fold increase in platelets and a l.5-fold increase in leukocyte concentration from the 1- to 1.2-mL plasma layer compared to baseline concentrations in whole blood. While almost no cells were found in the first 4-mL layer of L-PRF, a massive accumulation of platelets and leukocytes was found directly within the buffy coat layer demonstrating extremely high concentrations of cells in this 0.3-0.5-mL layer (~ 20-fold increases). We therefore proposed harvesting this 0.3- to 0.5-mL layer directly above the red blood cell corpuscle layer as liquid C-PRF. In general, i-PRF was able to increase platelet numbers by ~ 250%, whereas a 1200-1700% increase in platelet numbers could easily be achieved by harvesting this 0.3-0.5 mL of C-PRF (total platelet concentrations of > 2000-3000 × 109 cells/L). While conventional i-PRF protocols increase platelet yield by 2-3-fold and leukocyte yield by 50%, we convincingly demonstrated the ability to concentrate platelets and leukocytes over 10-fold by harvesting the 0.3-0.5 mL of C-PRF within the buffy coat following L-PRF protocols. Previous studies have demonstrated only a slight increase in platelet and leukocyte concentrations in i-PRF. The present study described a novel harvesting technique with over a 10-fold increase in platelets and leukocytes that can be further utilized for tissue regeneration.

Dominant negative Gfi1b mutations cause moderate thrombocytopenia and an impaired stress thrombopoiesis associated with mild erythropoietic abnormalities in mice.

GFI1B-related thrombocytopenia (GFI1B-RT) is a rare bleeding disorder mainly caused by the presence of truncated GFI1B proteins with dominant-negative properties. The disease is characterized by low platelet counts, the presence of abnormal platelets, a megakaryocytic expansion, and mild erythroid defects. However, there are no animal models that faithfully reproduce the GFI1B-RT phenotype observed in patients. We had previously generated mice with floxed Gfi1b alleles that can be eliminated by Cre recombinase, but those animals developed a much more severe phenotype than GFI1B-RT patients and were of limited interest in assessing the disease. Using CRISPR/Cas9 technology, we have now established three independent mouse lines that carry mutated Gfi1b alleles producing proteins lacking DNA binding zinc fingers and thereby acting in a dominant negative (DN) manner. Mice heterozygous for these Gfi1b-DN alleles show reduced platelet counts and an expansion of megakaryocytes similar to features of human GFI1B-RT but lacking the distinctively large agranular platelets. In addition, Gfi1b-DN mice exhibit an expansion of erythroid precursors indicative of a mildly abnormal erythropoiesis but without noticeable red blood cell defects. When associated with megakaryocyte-specific ablation of the remaining allele, the Gfi1b-DN alleles triggered erythroid-specific deleterious defects. Gfi1b-DN mice also showed a delayed recovery from platelet depletion, indicating a defect in stress thrombopoiesis. However, injecting Gfi1b-DN mice with romiplostim, a thrombopoietin receptor super agonist, increased platelet numbers even beyond normal levels. Thus, our data support a causal link between DN mutations in GFI1B and thrombocytopenia, and suggest that patients with GFI1B-RT could be treated successfully with thrombopoietin agonists.