DİCLE ÜNİVERSİTESİ TIP FAKÜLTESİ ARAŞTIRMA HASTANESİ ACİL SERVİSİNDE YAPILAN KAN VE KAN ÜRÜNLERİ TRANSFÜZYONLARININ DEĞERLENDİRİLMESİ

This study was designed prospectively to investigate the demographic data of the patients who had blood and blood products in Dicle University Medical Faculty Research Hospital Emergency Service between 01 November 2014- 01 November 2015, also to investigate the indications for blood products used, the amount and the type of the products used. When we look at the diagnosis of patients who underwent transfusion in the emergency department; It was determined that 26.2% were transfused for malignancy, 32.6% for anemia, 18.3% for trauma, 12.5% for GIS bleeding, and 10.4% for other reasons. The patients who had blood product transfusion were 623; 53.8% were male and 46.2% were female. The mean age of the patients was determined 47.87±23.66. The percentage of transfused blood products were as following: 83.6% erythrocyte suspension (ES), 17.3% fresh frozen plasma (FFP), 17.3 % platelet suspension(PS), 0.6% whole blood(WB), %4.3 other blood components (OBC). As a result, we can say that in our study, a large number of transfusions were performed in the emergency department, a specific protocol was not adhered to in determining the indication, and transfusion was performed in support of patients who were mostly under outpatient follow-up. Emergency transfusion decisions should be performed with true emergency indication, follow-up with clinical response rather than laboratory response, and consciousness about the aim of transfusion.

Serotonin and cyclic sleep organization in healthy full-term newborns

Relevance: The growth of neuropsychiatric diseases caused by perinatal pathology indicates the need to study the biochemical markers of brain damage in the newborn for the timely prevention of adverse consequences. Serotonin in early ontogenesis provides intensive development of neuronal structures and cortical networks involved in the mechanisms of formation of cyclic sleep organization a fine criterion of morphofunctional development of the brain. aim: The aim of the work is to study the content of serotonin in healthy full-term newborns in comparison with the quantitative and qualitative characteristics of the electropoligraphic sleep pattern. Material and methods: 84 healthy newborns were examined, which, depending on the gestational age, were divided into 3 groups: I 37 weeks (20 people), II 38 weeks (24 people), III 39-40 weeks (40 people). The content of serotonin in platelet-rich plasma of blood from the umbilical cord vein and in platelet suspension prepared from venous blood taken from mothers and children on the first day of life and again on day 5 was determined by high-performance liquid chromatography with electrochemical detection. A quantitative and qualitative analysis of the sleep electropoligram was performed 7-12 hours after birth. Results: The content of serotonin in platelet-rich plasma in umbilical cord blood in children does not depend on the method of birth, is 2 times lower than in the venous blood of mothers (0.379 0.116 microns/l, versus 0.756 0.200 microns/l, but there is a high correlation between the indicators (r = 0.8, p 0.05). At the gestational age of 39-40 weeks, the level of serotonin in platelet-rich plasma and in venous blood platelets is significantly higher than in those born at 37 weeks. In the latter, the increase in the content of serotonin in platelets continues after birth (at day 1, 0.539 0.149 nM/109 Tr, and on day 5 0.846 0.094 nM/109 Tr; p 0.05), whereas the indicators for those born at 39-40 weeks of pregnancy. They do not change (0.797 0.190 nM/109 Tr and 0.749 0.142 nM/109 Tr, respectively). A significant increase in the content of serotonin in the platelet-rich plasma and in the platelets of the child in the period from 37 to 39 weeks, both during intrauterine development and in the first days of life, correlates with an increase in the representation of the orthodox phase in the sleep cycle. Conclusion: The general pattern of changes in serotonin content and cyclic sleep organization in the early neonatal period in healthy newborns, depending on gestational age, indicates the possibility of using the obtained standard values of serotonin as a biochemical marker of functional brain development.

Use of increased concentrations of adenosinediphosphate for high residual platelet reactivity in patients with coronary heart disease

Detection of a tendency to increased thrombosis in patients with coronary heart disease (CHD) is of important prognostic value in the selection of drugs aimed at achieving a persistent antithrombotic effect. The aim of the study was to evaluate the use of elevated ADP inducer concentrations to improve the accuracy of ADP-induced platelet aggregation in patients with coronary heart disease. Material and method. Material and method. We studied 48 patients with CHD who were on continuous double antiplatelet therapy for 6 months (aspirin 75mg and clopidogrel 75mg per day). The aggregation activity of the platelet suspension was studied using the Born method G. in the modification of Gabbasov Z. A. Platelet activity was evaluated by the degree of aggregation of platelet-rich plasma along the light transmission curve under the influence of the inducer adenosine diphosphate (ADP) at a concentration of 2 mmol/l and by its own patented method against the background of additional ADP application. Results. In patients, platelet aggregation decreased to 5-35% (p<0.005) compared to the standard values, which are 50-60%. The values of platelet aggregation with the additional introduction of the inducer of aggregation ADP in a ratio of 2:1 to 2 µmol/l for 1, 2, 3, and 4-minute registration of platelet aggregation, resulted in increased aggregation from 55% to 75% (p<0.001), indicating high residual platelet reactivity on the background of double antiplatelet therapy. Correlations of the degree of aggregation for elevated ADP concentrations with multivessel arterial lesion and dyslipidemia were also found, r=0.86 and r=0.92, respectively. Conclusion. The use of elevated concentrations of adenosine diphosphate in platelet aggregation in patients with ischemic heart disease increases the accuracy of assessing ADP-induced platelet aggregation against the background of dual antiplatelet therapy and contributes to the detection of high residual platelet reactivity.

Anaesthesia for Caesarean Section of Primigravida with Idiopathic Thrombocytopenic Purpura

Idiopathic Thrombocytopenic Purpura (ITP) is a rare auto-immune disorder that causes low platelet levels. In this condition, platelets are prematurely destroyed by reticuloendothelial system. Here, authors report a case of 27-year-old primigravida of 38 weeks pregnancy with history of ITP was posted for elective caesarean section. Her platelet count was 25000/mm3 which increased to 78000/mm3, after treatment with 30 gm Intravenous (IV). immunoglobulin for three days and three units of pooled platelet suspension. Thereafter, caesarean section was conducted under general anaesthesia without any complication. Regional anaesthesia is a relative contraindication in obstetric patients with thrombocytopenia. To avoid the complications of neuraxial blockade, general anaesthesia was preferred and is safer in a patient with ITP during pregnancy with low platelet count.

Neonatal Alloimmune Thrombocytopenia Detected After Traumatic Delivery: Case Report

Neonatal Alloimmune Thrombocytopenia (NAIT), a complication of maternal alloimmunization against fetal platelets, is the result of immune destruction of platelets due to maternal antibodies in the early period. The cause of thrombocytopenia here is by antibodies developed against human platelet antigens frequently inherited from the mother. The clinical manifestations of NAIT extend from asymptomatic to severe bleeding. Platelet suspension can be used in the setting of severe thrombocytopenia or life-threatening bleeding. High-dose intravenous immunoglobulin (IVIG) therapy is another treatment option. A case who had no clinical finding except diffuse ecchymoses after traumatic delivery, but was considered to have NAIT due to severe thrombocytopenia and was successfully treated with platelet suspension and IVIG is presented.

Nonredundant Roles of Platelet Glycoprotein VI and Integrin αIIbβ3 in Fibrin-Mediated Microthrombus Formation

Objective: Fibrin is considered to strengthen thrombus formation via integrin αIIbβ3, but recent findings indicate that fibrin can also act as ligand for platelet glycoprotein VI. Approach and Results: To investigate the thrombus-forming potential of fibrin and the roles of platelet receptors herein, we generated a range of immobilized fibrin surfaces, some of which were cross-linked with factor XIIIa and contained VWF-BP (von Willebrand factor-binding peptide). Multicolor microfluidics assays with whole-blood flowed at high shear rate (1000 s −1 ) indicated that the fibrin surfaces, regardless of the presence of factor XIIIa or VWF-BP, supported platelet adhesion and activation (P-selectin expression), but only microthrombi were formed consisting of bilayers of platelets. Fibrinogen surfaces produced similar microthrombi. Markedly, tiggering of coagulation with tissue factor or blocking of thrombin no more than moderately affected the fibrin-induced microthrombus formation. Absence of αIIbβ3 in Glanzmann thrombasthenia annulled platelet adhesion. Blocking of glycoprotein VI with Fab 9O12 substantially, but incompletely reduced platelet secretion, Ca 2+ signaling and aggregation, while inhibition of Syk further reduced these responses. In platelet suspension, glycoprotein VI blockage or Syk inhibition prevented fibrin-induced platelet aggregation. Microthrombi on fibrin surfaces triggered only minimal thrombin generation, in spite of thrombin binding to the fibrin fibers. Conclusions: Together, these results indicate that fibrin fibers, regardless of their way of formation, act as a consolidating surface in microthrombus formation via nonredundant roles of platelet glycoprotein VI and integrin αIIbβ3 through signaling via Syk and low-level Ca 2+ rises.

Analysis of Pediatric Intensive Care Unit Admissions for Crimean–Congo Hemorrhagic Fever in Turkey

Objective The aim of this study was to determine the clinical characteristics, course of disease, and outcomes of patients admitted to our pediatric intensive care unit (PICU) for Crimean–Congo hemorrhagic fever (CCHF). Methods Data of all patients with CCHF who were admitted to PICU during a 3-year period and whose diagnosis was confirmed by serological methods or polymerase chain reaction were retrospectively reviewed. Their presenting characteristics, treatments, and outcomes were extracted from the hospital's electronic database. Disease severity scoring was performed using the severity scoring index and severity grading score system. Results The data of 22 of 123 patients with suspected CCHF who were admitted to PICU were evaluated. Among them, 68.2% were males and mean age of all patients was 13.4 ± 2.2 years. The average length of stay of 10 patients was >10 days. On average, 3.72 units of platelet suspension per patient was transfused. In the course of the disease, the highest laboratory values were reached on day 3 of admission. Conclusion Poor CCHF outcome depends on thrombocytopenia severity, prolonged coagulation tests, high-liver enzyme levels, and disseminated intravascular coagulation. Poor outcomes can be avoided by CCHF awareness and preparedness, early diagnosis of the disease, and supportive treatment with appropriate fluid and blood product transfusion. Randomized controlled trials on prophylactic transfusion and ribavirin use are needed.

Blood Product Requirement in Childhood Acute Lymphoblastic Leukemia in Relation to Chemotherapy Regimens

Background: Along with lack of studies that systematically address the transfusion requirements and triggers in acute leukemia, no study to date has addressed the blood product use with respect to ongoing chemotherapy (CT) in pediatric acute lymphoblastic leukemia (ALL) patients. Objectives: This study was therefore designed to evaluate total erythrocyte, random platelet and apheresis platelet suspension requirement in pediatric ALL patients in relation to ongoing CT protocols. Methods: A total of 146 pediatric patients with ALL were included in this retrospective study. Data on patient demographics, CT protocol, amount and efficacy of blood product use (erythrocytes, apheresis platelet and random platelet), and survival during CT were retrieved from hospital records. Results: The average amount of erythrocytes, apheresis platelets and random platelets received by 146 ALL patients from the date of diagnosis were 14 (3 - 78) bags, 9 (1 - 97 bags and 11 (1 - 83) bags, respectively. Protocol 1b augmented was associated with the highest amount of erythrocyte use (P < 0.001), while no significant difference was noted in apheresis platelet and random platelet use with respect to CT regimens. Erythrocyte transfusion was associated with a more marked increase in hemoglobin (Hb), erythrocyte (RBC), leukocyte (WBC), lymphocyte and neutrophil counts as compared with apheresis platelet and random platelet infusions, while protocol 2 was associated with higher Hb (P = 0.017) levels after erythrocyte transfusion. Conclusions: Our findings indicate a great amount of blood product transfusion to be required in children with ALL under CT and emphasize the likelihood of transfusion need and efficacy of transfusion to alter with respect to ongoing CT regimen. The need in patients using augmented BFM protocol 1 b was highest, and albeit the need of blood and blood product transfusions vary within patients, the anticipated median need for blood products at diagnosis and at various blocks of treatment may be helpful for the blood banks, doctors of the respective pediatric hematology-oncology centers to plan as patients are treated.

Novel Methods to Quantitate and Identity Dividing Platelets in Culture Conditions

Background: Unlike dividing cells, platelets are usually regarded as unproductive in that they are anuclear fragments of megakaryocyte and incapable of the canonical cell cycle. However, recent evidence showed that platelets could form progeny or at least undergo fission. The observation indicated the existence of a non-hematopoietic mechanism to determine platelet count in peripheral blood. The assessing the kinetic aspects of platelet division is not easy because automated cell counters often give equivocal results after culture not supporting those previous findings. Manual chamber counting is impractical because of the difficulty in differentiating singlets from any small sized clumps. Objective: We examined platelet division under in-vitro culture conditions to assess how many platelets and how fast they divide. Methods: We devised a method to track platelet division based on flow cytometry and fluorescent microscopic image. Briefly, buffered human platelet suspension was prepared and stained with cell tracking dyes followed by sorting and resuspension in cell culture medium and incubation. Platelet division was assessed quantitatively by observing a decrease in fluorescent intensity by flow cytometry after culture in a liquid medium or counting the number of single colored platelet doublets after culture in hydrogel medium. Results: After 6-hour culture of CFSE labelled platelet in M199 medium, we observed the appearance of a separate cluster of platelets with reduced CFSE intensity adjacent to the original platelet cluster, as is the typical finding of CFSE dilution assay for lymphocyte proliferation. We continued the culture for 20 hours. The division fraction of strongly labeled platelets was 48.7 % (range: 27.5 - 70.4) after 6 hours and 85.6 % (79.1 - 92.0) after 20 hours. For weakly labelled platelets, the division fraction after 6 hours was 35.0 % (24.6 - 43.4) and 75.5 % (34.5 - 97.0) after 20 hours. The division fractions at the two time points were much higher than expected and the effect of external stimuli such as shearing force exposure during the sorting procedure was to be considered. In another assay, we divided platelet suspension into two and stained with labelling dyes of different colors (CFSE and FarRed). The two fractions were mixed and cultured in hydrogel medium. At the beginning of the culture and after 6 hours, we counted platelet singlets and doublets in ten randomly selected fields of con focal microscopy. Single colored doublets can form both by division and contact. Those with two colors can form only by contact. The count of single- and two-colored doublets can be estimated by binary probability prediction if the platelets don't divide. Initially, the doublet counts follow the binary probability prediction (P: 0.65 - 0.88), but after 6 hours, single colored doublets were observed more frequently than two colored doublets (P<0.01, in the order of 10-6). The division was suppressed by treatment with taxol (P =0.26), nocodazole (P=0.38), and cytochalasin D (P=0.61). From the counted numbers of doublets and singlets of the two colors, we could derive a formula for the platelet division fraction. The division fraction of FarRed stained platelets were 8.3 % (95% CI: 2.2 - 14.4), and that of the CFSE stained platelets were 11.9 % (5.4 - 18.4). Conclusion: We introduce novel methods to assess platelet division credibly in culture conditions which can be easily combined with functional studies. Figure Figure. Disclosures No relevant conflicts of interest to declare.

Platelet-Rich Plasma in Tissue Engineering: Hype and Hope

Background: Platelet-rich plasma (PRP) refers to an enriched platelet suspension in plasma. In addition to the clinical application of PRP in the context of various orthopedic diseases and beyond, PRP and platelet lysate (PL) have been in focus in the field of tissue engineering. In this review, we discuss the application of PRP as a cell culture supplement and as part of tissue engineering strategies, particularly emphasizing current hurdles and ambiguities regarding the efficacy of PRP in these approaches. Summary: As a putative autologous replacement for animal-derived supplements such as fetal calf serum (FCS), PRP has been applied as cell culture supplement for the expansion of stem and progenitor cells for tissue engineering applications and cell therapies. Attributed to the high content of growth factors in platelets, PRP has been shown to promote cell growth, which was mostly superior to standard cultures supplemented with FCS, while the differentiation capacity of progenitor cells seems not to be affected. However, it was also suggested that cultivation of cells with PRP significantly alters the protein expression profile in cells in comparison to FCS, indicating that the influence of PRP on cell behavior should be thoroughly investigated. Moreover, different PRP preparation methods and donor variations have to be considered for the use of PRP under good manufacturing practice conditions. PRP has been used for various tissue engineering applications in the context of bone, cartilage, skin, and soft tissue repair, where most studies were conducted in the field of bone tissue engineering. These approaches take either advantage of the release of chemoattractive, angiogenic, proliferative, and putatively pro-regenerative growth factors from PRP, and/or the hydrogel properties of activated PRP, making it suitable as a cell delivery vehicle. In many of these studies, PRP is combined with biomaterials, cells, and in some cases recombinant growth factors. Although the experimental design often does not allow conclusions on the pro-regenerative effect of PRP itself, most publications report beneficial effects if PRP is added to the tissue-engineered construct. Furthermore, it was demonstrated that the release of growth factors from PRP may be tailored and controlled when PRP is combined with materials able to capture growth factors. Key Messages: Platelet-derived preparations such as PRP and PL represent a promising source of autologous growth factors, which may be applied as cell culture supplement or to promote regeneration in tissue-engineered constructs. Furthermore, activated PRP is a promising candidate as an autologous cell carrier. However, the studies investigating PRP in these contexts often show conflicting results, which most likely can be attributed to the lack of standardized preparation methods, particularly with regard to the platelet content and donor variation of PRP. Ultimately, the use of PRP has to be tailored for the individual application.