Sodium Citrate Anticoagulant Research Articles

Intervention measures to improve the filter life of continuous renal replacement therapy in critically ill patients-A systematic review.

Continuous renal replacement therapy (CRRT) is a method of blood purification, which is widely used in the treatment of critical diseases as a means of multiple organ function protection and life support therapy. However, because of the serious condition of ICU (intensive care unit) patients, CRRT needs to be carried out continuously, but the treatment is interrupted ahead of time as a result of various conditions, which not only affects the treatment effect but also increases the patient cost. To evaluate intervention measures to improve the filter life of CRRT in critically ill patients, and also to identify which interventions are considered 'promising interventions'. This is a systematic review. Seven databases were searched using terms related to the concepts of 'continuous renal replacement therapy' and 'filter life', from the establishment of the database to 31 December 2023. The quality of the methodology included in the study was assessed using standard evaluation tools developed by the Effective Public Health Practice Project (EPHPP), and pre-established criteria were used to identify 'promising interventions'. A total of 28 studies were included, of which 7 were rated 'strong' in terms of design and methodological quality, and the others were 'medium'. The most commonly identified interventions to extend the life of CRRT filters include the use of sodium citrate anticoagulation, the choice of CVVHD or CVVHDF or pre-diluted CVVH for CRRT and the use of personalized sodium citrate anticoagulant regimens to reduce the incidence of filter clotting. The intervention measures of 14 studies were statistically significant, while the other 14 studies were not statistically significant. Interventions in nine studies were identified as 'promising interventions' because they were published within 10 years, with a medium or strong methodological quality rating, significant positive results and a strong evidence base. In the promising interventions study, citrate anticoagulation and CVVHD or CVVHDF models were recommended to significantly prolong filter life. However, more high-quality studies are needed to identify interventions that can prolong the life of CRRT filters in critically ill patients, thereby supplementing the literature in this field. The existing studies lack blinding and have limited quality. Future studies should be carried out with the goal of 'best evidence', and the interventions should be more universal and clinically practical. This study uses the method of systematic review to scientifically and rigorously provide some suggestions for extending the life of CRRT filters in critically ill patients, such as 'By implementing personalized citrate anticoagulation protocols, the incidence of CRRT filter life shortening due to coagulation can be reduced', which can reduce the cost of patients and improve the quality of treatment to a certain extent in clinical practice.

Inadequate anticoagulation and hyperuricemia cause knee pain after platelet-rich plasma injection: A retrospective study.

Platelet-rich plasma treatment delays the need for total knee replacement in patients with knee osteoarthritis. However, its use and preparation remain controversial. The aim of this study was to investigate the relationship between anticoagulant use in the preparation of platelet-rich plasma and post-treatment pain in patients with knee osteoarthritis. Additionally, we explored the efficacy of platelet-rich plasma over medium- and long-term follow-up periods and identified other factors that may affect treatment outcomes. In this retrospective study, 225 patients with knee osteoarthritis, who underwent knee platelet-rich plasma treatment from June 2021 to January 2022, were examined at three study centres. Patients were categorised, based on the type and amount of anticoagulant used during platelet-rich plasma preparation, into 4% sodium citrate (SC) 0.6mL, 4% SC 1mL, 4% SC 2mL, heparin 0.1mL, and heparin 0.2mL groups. We analysed the patients' basic information, pain after treatment, and inflammatory markers (i.e., interleukin 6, tumour necrosis factor-α, and hypersensitive C-reactive protein) in the joint fluid via enzyme-linked immunosorbent assay and joint fluid crystallisation. Additionally, we assessed the patients' Western Ontario and McMaster University scores and minimal clinically significant differences after treatment. Patients in the 4% SC 0.6mL and heparin 0.1mL groups experienced less pain after platelet-rich plasma treatment than did patients in the high-dose anticoagulant group. The joint fluid of patients with pain in these groups had lower levels of inflammatory markers. Patients treated with SC had slightly better medium- and long-term therapeutic outcomes than did patients treated with heparin. Patients with poorly controlled hyperuricemia also experienced pain after platelet-rich plasma treatment. The results suggest that platelet-rich plasma prepared using high-dose anticoagulants or administered to patients with poorly controlled hyperuricaemia may lead to moderate-to-severe knee pain and joint effusion after joint puncture therapy. Platelet-rich plasma had a therapeutic effect on knee osteoarthritis; however, its efficacy gradually decreased over time. SC anticoagulant is more suitable for platelet-rich plasma preparation than is heparin. Further studies are needed to understand the safety and the various factors influencing platelet-rich plasma therapy.

Filter Lifespan, Treatment Effect, and Influencing Factors of Continuous Renal Replacement Therapy for Severe Burn Patients.

Continuous renal replacement therapy (CRRT) is often disrupted due to various factors, such as patient-related issues, vascular access complications, treatment plans, and medical staff factors. This unexpected interruption is referred to as non-selective filter stoppage and can result in additional treatment expenses. This study conducted a retrospectively analyzed 501 CRRT filters used in 62 patients with severe burns, lifespan and therapeutic effect of all filters were mainly analyzed, used logistic regression analysis was performed to identify risk factors associated with non-selective cessation filters. Out of 493 filters, 279 cases received heparin (56.60%), the median lifespan of the filter was 14.08 h (25th, 75th quantile: 7.30, 21.50); 128 cases were treated with nafamostat mesylate (26.00%), and the median lifespan of the filter was 16.42 h (10.49, 22.76); 86 cases were treated with sodium citrate (17.40%), and the median lifespan of the filter was 31.06 h (19.25, 48.75). In addition, significant differences were observed in the electrolyte index, renal function index, and procalcitonin levels before and after treatment with a single filter (P < .001). Multivariate logistic regression showed that the risk of non-selective cessation of sodium citrate anticoagulants was lower than that of heparin anticoagulation. Overall, CRRT is progressively becoming more prevalent in the treatment of patients with severe burns. The lifespan of individual filters and total patient treatment duration showed a consistent upward trend. The filter's lifespan was notably greater during sodium citrate anticoagulation when compared to nafamostat mesylate and heparin, meanwhile notably reducing the risk of non-selective cessation. Therefore, we recommend sodium citrate for anticoagulation in patients without any contraindications.

Variations in the full blood count parameters among apparently healthy humans in the Ho municipality using ethylenediamine tetraacetic acid (EDTA), sodium citrate and lithium heparin anticoagulants: A laboratory-based cross-sectional analytical study

BackgroundSeveral studies have shown that various anticoagulants used for collection of blood samples produce varying effects on haematological analyses. Tripotassium ethylenediamine tetra-acetic acid (K3EDTA), sodium citrate and lithium heparin remain the most used anticoagulants employed in hematological analysis. There is paucity of data on the effect of these anticoagulants on haematological parameters in humans in Ghana. We assessed the suitability of K3EDTA, sodium citrate and lithium heparin for routine Full Blood Count (FBC) investigation. MethodA laboratory-based analytical cross-sectional study was conducted using blood samples from 55 conveniently sampled apparently healthy tertiary students from January 2021 to October 2021. Blood samples were taken from each participant into 3 anticoagulant tubes: K3EDTA, sodium citrate and lithium heparin and FBC parameters estimated using the Mindray automated haematology analyzer. One-way ANOVA Kruskal-Wallis test, Mann-Whitney U, Intra-class correlation coefficient (ICC) analysis, Bland-Altman's plot and Lin's concordance correlation coefficient were used where appropriate to ascertain the level of variation, consistency, and agreements among and between results. Normality testing using Shapiro-Wilk test statistic revealed non-Gaussian distribution of data, hence, were presented as median, minimum, and maximum. Data generated were analyzed using STATA v15 and MedCalc v20 where appropriate for statistical analysis. P-values <0.05 were considered statistically significant. ResultsThe study comprised 34 males and 21 females. The median age for males (23 years: min = 20, max = 34) was statistically comparable (p = 0.2652) to that of females (22 years: min = 18, max = 34). We observed excellent consistency in the estimation of MCV (ICC = 0.94), MCH (ICC = 0.98), MCHC (ICC = 0.91), GRAN# (ICC = 0.92) and LYMPH% (ICC = 0.91) across the three anticoagulants. Heparin and K3EDTA largely agreed on most of the FBC parameters, 50.0% (7/14) including HGB, MCV, MCH, PLT, LYMPH#, GRAN# and GRAN%. Meanwhile using K3EDTA as a standard, heparin produced almost perfect agreement only in the assessment of RBC (CCC = 0.992) while a substantial agreement was observed in the assessment of HGB (0.971), HCT (0.958) and MCH (0.987). Citrate agreed substantially with K3EDTA in the assessment of LYMPH% (CCC = 0.964) and moderately in the assessment of MCV (CCC = 0.948) and MCH (0.913). Overall, compared to K3EDTA, heparin was highly precise and accurate in the estimation of HGB, RBC, HCT and MCH while citrate determined MCV and MCH more accurately and precisely. ConclusionCitrated blood consistently produced lower FBC values compared to heparin and K3EDTA and hence suggests not reliable in the assessment of FBC among humans. Heparin agreed largely with K3EDTA in the estimation of FBC parameters and may be used as a better alternative anticoagulant in the absence of K3EDTA however with great caution.

Influence of Heparin on Fibrinogen Assay by Clauss Method

Objective: To investigate the effect of heparin on fibrinogen detection by Clauss method. Methods: A normal plasma pool (NPP) of 20 healthy people was prepared with 3.2% sodium citrate anticoagulant. For experimental group 1, samples containing different concentrations of heparin were prepared. For experimental group 2, samples of experimental group 1 were diluted twice. For experimental group 3, samples of experimental group 1 were diluted 4 times. For the control group, samples of normal saline with the same volume as heparin in experimental group 1 were prepared. The fibrinogen contents of experimental group 1, experimental group 2, experimental group 3, and control group were detected by Clauss method and prothrombin time (PT)-derived method, and the fibrinogen detection results of different groups were analyzed. Results: The trend of fibrinogen detected by Clauss method and PT-derived method in experimental group 1 was different; there was significant difference between the results of experimental group 1 and the control group (P &lt; 0.05); there was no significant difference between the results of experimental group 2 and the control group (P &gt; 0.05); there was no significant difference between the results of experimental group 3 and the control group (P &gt; 0.05); there was no significant difference between the results of experimental group 2 and experimental group 3 (P &gt; 0.05); the relative deviation between experimental group 1 and the control group was higher in high-concentration heparin sample. Conclusion: Heparin affects fibrinogen detection by Clauss method, and the effect can be reduced by sample dilution.

Evaluation of sodium citrate anticoagulant for the Resolution of edta-dependent pseudo Thrombocytopenia.

EDTA-dependent pseudo thrombocytopenia (EDTA-PTCP) refers to a falsely low platelet count occurring in the presence of ethylene diamine tetra-acetic acid (EDTA) anticoagulant during blood sample collection, which results in the formation of platelet clumps in vitro. This phenomenon has significant clinical implications, including unnecessary administration of platelets. Our study aims to evaluate the efficacy of sodium citrate anticoagulant for the resolution of EDTAPTCP. This retrospective study was conducted in the haematology laboratory of Shifa International Hospital (SIH), Pakistan. Patients with pseudo thrombocytopenia (i.e. platelet count less than 150,000/ul with platelet clumps seen on peripheral smear) were included in this study if they had blood samples drawn in both EDTA and sodium citrate tubes less than 48 hours apart. Data was analyzed using IBM® SPSS Software Version 22. A total of 151 study participants were included in this study. The mean age was 48.95±20.69 years and the majority were female (52.3%). Wilcoxon signed-rank test showed that there was a statistically significant difference in platelet count measured in both tubes (Z = -3.223, p=0.001). Overall, blood samples processed in sodium citrate tubes showed lower platelet count than EDTA samples. Sodium citrate anticoagulant was able to correct EDTA-PTCP in 47 (31.1%) of the cases. Sodium citrate anticoagulant was only able to resolve one-third of our EDTA-PTCP cases. Our findings do not support the use of sodium citrate as a suitable alternative for correction of EDTA-PTCP.

Platelet Aggregation in Apparently Healthy Elderly Populations: Reference Ranges and Influence of Hematology Parameters.

Reference intervals based on younger populations may not apply to elderly populations. The aim of the current study was to calculate reference ranges for platelet aggregation in apparently healthy Chinese elderly populations and analyze the impact of gender, age, and hematological parameters on platelet aggregation in vitro. A total of 138 males and 161 females were enrolled based on stringent inclusion and exclusion criteria. Platelet aggregation was measured with sodium citrate anticoagulation whole blood samples using a PL12 analyzer, triggered by adenosine-5'-diphosphate (ADP) and arachidonic acid (AA) agonists. Hematological parameters were measured using a Sysmex XE2100 instrument. In the total sample tested in this study, the platelet aggregation induced by AA and ADP was not dependent on age (p > 0.05) but gender. Platelet aggregation triggered by ADP and AA was more enhanced in females than males (p = 0.024 for AA, p = 0.036 for ADP). The recommended reference values of AA-induced platelet aggregation were 48.42% - 85.93% for males and 43.62% - 87.80% for females. The recommended reference values of ADP-induced platelet aggregation were 38.12% - 80.21% for males and 40.12% - 83.05% for females. Furthermore, for all agonists, a positive correlation was found between platelet aggregation and platelet count. The ADP-triggered aggregation showed a significant positive correlation with white blood cell (WBC) count and a negative correlation with red blood cell (RBC) count. Moreover, platelet aggregation showed no significant correlation with mean platelet volume or hemoglobin concentration. Combined and gender-specific reference ranges for platelet aggregation in healthy elderly populations were established in whole blood measured by a sequential platelet counting method.

Comparison of Complete Blood Count Parameters using EDTA, Sodium Citrate, and Heparin Anticoagulants

A complete blood count is a routine blood examination performed in laboratories. EDTA is the standard anticoagulant used in complete blood count examination. The use of ethylenamineintetraacetic acid (EDTA) may cause pseudo thrombocytopenia. The alternative to EDTA is sodium citrate and heparin anticoagulants. This study aimed to compare the results of complete blood count parameters using EDTA, sodium citrate, and heparin anticoagulants for the necessity of health service in the laboratory. The design of this study was a cross-sectional observational analytic done from October 2020 until November 2020. The participants of this study came from 62 adults who look healthy recruited in Medical Check-up Department. The sample was collected using K2- EDTA, 3.2% sodium citrate, and heparin tubes with volumes of 3ml, 3ml, and 4ml respectively. The sample was then examined using Sysmex XN 1000 hematology analyzer. There were no significant differences between the result of hemoglobin (HBG), red blood cells (RBC), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), red cells distribution width (RDW), and white blood cells (WBC) parameters. The result of platetelet (PLT) was significantly different between the three tubes. The results of platelet examination with sodium citrate and heparin tubes were lower than those obtained with the EDTA tube. EDTA is still the best choice for a complete blood count, but sodium citrate and heparin anticoagulants can be used as alternatives for testing.

Measurement of Protein C Levels among Sudanese Children and Infants with Sepsis at Khartoum state, 2022

Background: Sepsis is a life-threatening condition that arises when the body's response to infection causes injury to its own tissues and organs.&#x0D; Materials and methods: This was cross sectional study conducted at Baraah pediatric center, Khartoum, Sudan during the period from May 2022 to August 2022, to measure protein C level in Sudanese children and infants with Sepsis. 50 patients were selected as a case group (46% were female and 54% were male.), and apparently healthy donors were selected as control group. 2.8 ml of venous blood samples were collected in Tri Sodium Citrate Anticoagulant. CRP and the blood culture were done for all the patients. The protein C level was performed using Ichroma device.&#x0D; Results: Protein C results revealed the mean of protein C in case was (63.7±8.9 ), and in the control group was(79.8±11.1 ), when compared protein C mean between case and control groups there was a highly significant decreased with ( p value 0.00) Also in the case group there was an insignificant differences between protein C, age, gender, and culture, p value &gt;0.05), . In addition the result showed insignificant correlation between protein C and CRP(p value ≥0.05). The most common types of isolated bacteria in sepsis patients was Klebsiella&#x0D; Conclusion: The study concludes that there is a significant decrease in protein C level in patients with sepsis.&#x0D; Keywords: protein C , CRP sepsis , bacteria , Klebsiella

Evaluation of Complete Blood Count and D. Dimer in Patients with COVID-19 Infection in Shendi Town

Background: Covid-19 can Cause Various Conditions Including respiratory, enteric, and neurological diseases, and led to a pandemic that has affected millions worldwide. Methods: This is a cross-sectional descriptive study conducted at Shendi teaching hospital which is located in Shendi town in Sudan to evaluate Haematological parameters and D. Dimer in patients with Covid-19 Infection in the period between May to September 2021. The study included (50) patients who were diagnosed with Covid-19 Infection and the study groups were compared with (50) healthy volunteers as a control group. 50 venous blood samples as case and 50 as control transferred into Tri sodium citrate and EDTA anticoagulant. Data was collected using a questionnaire and the (SPSS) version (22) program was used for data analysis. Results: The study revealed that the Covid-19 Infection patients were; (60%) males and (40) females. Complete blood count (CBC) indicated the mean values of Hb, PCV, RBCs, MCV, MCH, MCHC, in case group were (12.5 g/dl), (38.9%), (4.5x1012/l), (86.7 fl), (27.7 pg) and (32.0 g/dl) respectively. Also prevailed the mean of TWBCs, Neutrophil, lymphocyte, MID, Absolute Neutrophil, Absolute lymphocyte, Absolute MID, were (13.09x 109 /l), (79.27%), (11.79%), (8.77%), (10.38), (1.54), and (1.16) respectively. The study revealed the mean of platelet, D.dimer (296.63x 109 /l), (6.07ug/ml), respectively. Conclusions: Covid-19 Infection is responsible for significant changes in hemoglobin, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration, total white blood cells count differential and absolute neutrophils count and differential and absolute lymphocytes count and D. dimer.

Measurement of Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT), Fibrinogen Level, D-Dimer in Sudanese Infants and Children with Sepsis Khartoum State, 2022

Background: Sepsis is the most common worldwide cause of death in infants and children. It was Known that coagulative and inflammatory parameters were involved in the host’s defense against sepsis resulted from severe infection and inflammation. Materials and Methods: This was cross sectional study conducted at Albraa pediatric center, Khartoum, Sudan during the period from May 2022 to August 2022, to measure PT, APTT, fibrinogen level, D- dimer in Sudanese infants and children with sepsis. 50 patients were selected as a case group, and apparently healthy donors with no history of any coagulation problems or any chronic disease were selected as control group. 2.8 ml of venous blood samples were collected in Tri Sodium Citrate anticoagulant. The coagulation tests (PT, APTT, fibrinogen) were performed using semiautomatic device (coagulometer). I chroma was used for the D- dimer. Results When compared between case and control there was highly significant difference (p.value 0.000), in PT, APTT, fibrinogen and D- dimer. In addition, there was insignificant difference when the PT, APTT fibrinogen and D- dimer in cases compared with the gender, CRP and bacterial culture, and there was negative correlation between the parameters and the age except the D-dimer had appositive correlation. The most common types of isolated bacteria in sepsis patients is staphylococcus aureus. Conclusion: This study concluded that, PT, APTT, fibrinogen and D- dimer was significantly increased in patients with sepsis.

Comparison of different modalities of continuous renal replacement therapy with regional sodium citrate anticoagulation in paediatric patients.

To evaluate the efficacy and safety of continuous renal replacement therapy (CRRT) modalities with regional sodium citrate anticoagulation (RCA) in children. This retrospective study was conducted at the paediatric intensive care unit of Hunan Children's Hospital in China. Medical records of paediatric patients hospitalised for RCA-CRRT between April 2017 and March 2021 were reviewed. Patients received continuous venovenous haemodialysis, continuous venovenous haemofiltration (CVVH), or continuous venovenous haemodiafiltration (CVVHDF). Patients on continuous venovenous haemodialysis (n = 2) were excluded because of their small sample size. The remaining participants were divided into CVVH and CVVHDF groups; 80 patients received CRRT, with 40 and 62 sessions in the CVVH and CVVHDF groups, respectively. The filtre lifespan was longer in the CVVHDF group than in the CVVH group (median value [interquartile range]; 47 [15] hours vs. 35 [17.5] hours; p = 0.029). Compared with the CVVHDF group, the hazard ratio for filtre lifespan in the CVVH group was 3.023 (95% confidence interval 1.820-5.023, p < 0.001). There were no significant differences in ionised calcium levels of the circuits between the two groups at different time points (p < 0.05). Metabolic alkalosis, hyperlactataemia, hypocalcaemia, and hypercalcaemia occurred in both groups, with metabolic alkalosis being the most common complication. No patients in either group experienced sodium citrate accumulation or hypernatraemia. Inter-group differences in the incidence of these complications were not statistically significant (p > 0.05). Our results suggest that CVVHDF is a better option for RCA-CRRT than CVVH.

Diagnostic Value of D-Dimer and INR in Patients Suspected to Have Prosthetic Valve Dysfunction.

Prosthetic valve dysfunction is a potentially critical complication of heart valve replacement. An easy and quickly applicable diagnostic procedure is required for recognizing the prosthetic valve dysfunction. The purpose of this study was to prospectively define the diagnostic value of D-dimer and INR level in predicting prosthetic valve dysfunction. This cross-sectional study was performed in 70 patients suspected to have prosthetic valve dysfunction admitted to Imam Ali Hospital, affiliated with Kermanshah University of Medical Sciences (KUMS), Kermanshah Province, Iran. Cinefluoroscopy, as the gold standard diagnostic test, was used for the diagnosis of prosthetic valve dysfunction in enrolled patients. Two milliliters of blood from each patient were taken into a tube containing sodium citrate anticoagulant. To evaluate D-dimer, the cutoff value was set at 500 ng/ml. Also, to evaluate international normalized ratio (INR), the cutoff value was set at 2. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR), and negative likelihood ratio (NLR) of the serum markers were used to describe predictive properties. Of 70 patients, 27 (38.6%) were male and 43 (61.4%) were female, and the mean age was 54.67±15.11 years (range, 18 to 80 years). Of 70 patients, 27 (38.6%) had prosthetic heart valve malfunction demonstrable by fluoroscopy, and 19 patients (27.1%) had D-dimer levels &gt;500 ng/ml. Elevated D-dimer levels (&gt;500 ng/ml) have been indicated to have sensitivity of 70.4%, and hence an NPV of 84.3%, specificity of 100%, PPV of 100%, NLR of 0.3, and the infinity value of PLR for predicting prosthetic valve dysfunction. There was a significant relationship between fluoroscopy and D-dimer test (P=0.001). A kappa coefficient value of 0.745 indicated a substantial agreement between D-dimer and fluoroscopy testing. Mixing test (combination of D-dimer and INR) showed to have 100% sensitivity, and hence a NPV of 69.8%, specificity of 69.8%, PPV of 51.8%, NLR of 1.41, and PLR of 1.44 for predicting prosthetic valve dysfunction. D-dimer with moderate sensitivity and high specificity is an ideal marker for the diagnosis of prosthetic valve dysfunction in suspected patients. Enhanced plasma D-dimer level is not by itself diagnostic of a prosthetic valve dysfunction but may alert physicians to refer the patient for more detailed examination, preferably by fluoroscopy. Mixing test with 100% sensitivity can apply as a rule-out test.

Effect of Time and Haemolysis on Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT) Measurement on Blood Samples

Background: Coagulation, also known as blood clotting, is the process by which blood convert from a liquid to a gel, forming a blood clot. It referred to haemostasis, the stopping of blood loss from a damaged vessel, followed by repair.&#x0D; Material and methods: This was cross sectional study conducted at the albawasla medical laboratory, Khartoum, Sudan during the period August to November, 2021 and to evaluate the effect of time and hemolysis on prothrombin time and activated partial thromboplastin time tests. 50 samples (case group) were collected from the patients attending police teaching hospital and requested to the PT and APTT test in addition to that,50 apparently healthy donors with no history of any coagulation problems or any chronic disease were selected as control group. Three ml of venous blood samples were collected in container with Tri Sodium Citrate anticoagulant. The coagulation tests (PT and APTT) were performed using semiautomatic device (coagulometer machine MI).&#x0D; Results: The result of this study revealed that; when compared the measurement of PT and APTT immediately and after one hour there was insignificant differences (p. v.&gt;0.05). also when compared the measurement of PT and APTT between hemolyzed and non-hemolyzed samples there was significant differences ( p. v.&lt;0.05) in addition when compared case and control for the PT and APTT immediately, after one hour, hemolyzed and non-hemolyzed sample there was significant differences ( p. v.&lt;0.05) except the APTT hemolyzed samples and insignificant differences with age and gender ( p. v.&gt;0.05). For the correlation there was significant correlation in the case group for the PT and APTT immediately, after one hour, and hemolyzed samples.&#x0D; Conclusion: In the cases group results showed insignificant differences in the results of PT and APTT between immediate sample and after 1 hour in and significant differences in the results of PT and APTT between hemolyzed and non-hemolyzed samples, also there was insignificant differences between age and gender, immediately, after one hr. and hemolyzed sample in PT and APTT.&#x0D; Keywords: Homeostasis, hemolyzed sample, PT and APTT

OPTIMIZING A PLATELET-RICH PLASMA CONCENTRATION PROTOCOL FOR SOUTH AMERICAN SEA LIONS (OTARIA FLAVESCENS).

Accelerated healing in wild or captive South American sea lions (Otaria flavescens) is a key tool to help minimize infection and complications associated with open wounds, dental disease, and ocular pathology. Platelet-rich plasma (PRP) is an autogenous source for growth factors based on platelet concentration, which can be obtained by centrifuging whole blood collected in sodium citrate anticoagulant. Currently, there are well-defined PRP concentration protocols for humans and most domestic companion animal species. However, there is no clear centrifugation protocol for obtaining PRP in most marine mammal species. This study aimed to optimize the platelet concentration protocol based on whole blood centrifugation using speeds ranging from 500 to 5,000 rpm and times ranging from 3 to 6 min. Blood was drawn from seven adult South American sea lions, placed into 1-ml sodium citrate tubes, and centrifuged following 12 different centrifugation protocols. PRP was designated as the lower third fraction of the centrifuged plasma. Platelet counts were performed using flow cytometry and statistical analysis was carried out to establish a well-defined protocol for efficient PRP production. Transmission electron microscopy (TEM) analysis was performed to evaluate possible platelet degranulation during the different centrifugation protocols and measure platelet areas. Maximum concentration of platelets in PRP was 4.73-fold higher than the number of platelets in equal volume of whole blood, and significant differences in the concentrations obtained were found between the 12 centrifugation protocols evaluated using different speed and time combinations. The best one-step centrifugation protocol resulted from using 900-rpm speed for 3 min. The highest-fold increase was achieved using a two-step centrifugation protocol, which combined the most efficient one-step centrifugation protocol (900 rpm, 3 min) with a second centrifugation using 2,000-rpm speed for 6 min. TEM analysis confirmed that platelets were complete and maintained integrity after the proposed protocol.

Assessment of a Novel Dielectric Microsensor for Monitoring Coagulation Factor Therapy in Children with Hemophilia with and without Inhibitors

Introduction: Monitoring coagulation factor or bypassing agent therapy has important clinical implications for the management of children with hemophilia, especially those with inhibitors. For patients with hemophilia, conventional laboratory-based coagulation tests are time-consuming, labor-intensive, and costly. Moreover, no monitoring assay is currently available for use with bypassing agents in patients with hemophilia and inhibitors, except for thromboelastography or rotational thromboelastometry (ROTEM), both of which are not widely available clinically and are labor-intensive. There is a need to develop an easy-to-use, low-cost, portable global assay of coagulation for monitoring therapies in hemophilia. The aim of this study was to assess the clinical utility of monitoring replacement therapy using a new technique based on a novel dielectric microsensor (ClotChip) that can potentially enable global blood coagulation assessment in a miniaturized, portable measurement platform.Methods: Patients accrued for the study were children with severe hemophilia A & B (n = 6), children with hemophilia and inhibitors (n = 4), and normal, healthy children without coagulopathy (n = 22). Blood samples were obtained by venipuncture into collection tubes containing 3.2% sodium citrate anticoagulant. Samples from patients with hemophilia were collected both prior to the initiation of replacement therapy and 15 minutes after administration of an appropriate coagulation factor VIII or IX for severe hemophilia A or B, respectively, or a bypassing agent (FEIBA or recombinant factor VIIa) for hemophilia with inhibitors. Whole blood samples were then tested with the ClotChip within 2 hours of collection, and coagulation was induced with CaCl2 prior to the start of ClotChip measurements. The ClotChip is based on the fully electrical technique of dielectric spectroscopy (DS), and features a three-dimensional, parallel-plate, capacitive sensing structure integrated into a low-cost (material cost < $1), small- sized (26mm × 9mm × 3mm), and disposable microfluidic biochip with miniscule sample volume (< 10µL). The ClotChip curves were calculated as the real part of blood permittivity at 1MHz, and the time to reach a peak in permittivity (Tpeak, Fig 1A) was taken as an indication of coagulation time. Samples from 8 of the 10 hemophilia patients were also tested with ROTEM using the non-activated TEM (NATEM) test according to the manufacturer's direction. The ROTEM clotting time (CT) parameter (Fig 1B) was used in this study as a clinically important parameter to monitor patients with hemophilia.Results: We observed a significant decrease in the ClotChip Tpeak parameter for post-therapy samples compared to baseline (pre-therapy) for children with hemophilia without inhibitors (Fig 1C, n = 6, p = 0.0001) and with inhibitors (Fig 1D, n = 4, p = 0.0024). Interestingly, baseline ClotChip measurements for 2 tolerized patients with inhibitors exhibited Tpeak of 970s and 1,010s, which are within the range of Tpeak for patients without inhibitors (750s - 1,320s), while the baseline ClotChip measurements of 2 non-tolerized patients with inhibitors exhibited higher Tpeak of 1,810s and 2,100s (Fig 1E). Moreover, ClotChip measurements for all samples taken post-therapy exhibited Tpeak that fell within the reference range of Tpeak for normal, healthy children (180s - 720s, Fig 1E). Taken together, these data suggest that the ClotChip Tpeak parameter is sensitive to detection and correction of coagulopathy in children with hemophilia with and without inhibitors, as indicated by Tpeak reaching reference range levels after coagulation factor therapy. Finally, the ClotChip Tpeak exhibited a very strong correlation (Fig 1F, Pearson's r = 0.94) to the clinically important CT parameter of ROTEM.Conclusions: Our studies demonstrate the feasibility of monitoring coagulation factor therapy in children with hemophilia with and without inhibitors, using a novel, microfluidic, dielectric sensor (ClotChip), allowing whole blood assessment of hemostasis in a single disposable sensor. ClotChip has potential to fulfill an important unmet clinical need for monitoring therapy, especially for hemophilia with inhibitors. Further studies are underway to assess the ability of ClotChip to differentiate bleeding phenotypes and monitor outcomes for treatment of bleeds in children with hemophilia. [Display omitted] DisclosuresAhuja:CSL Behring: Honoraria; Bayer: Honoraria; Shire: Honoraria; Shire: Speakers Bureau. Suster:XaTek, Inc.: Patents & Royalties: Michael Suster is an inventor of intellectual property that has been licensed by Case Western Reserve University to XaTek, Inc., and as such, may receive future royalties. Maji:XaTek, Inc.: Patents & Royalties: Denath Maji is an inventor of intellectual property that has been licensed by Case Western Reserve University to XaTek, Inc., and as such, may receive future royalties. Mohseni:XaTek, Inc.: Patents & Royalties: Pedram Mohseni is an inventor of intellectual property that has been licensed by Case Western Reserve University to XaTek, Inc., and as such, may receive future royalties.

Platelet and leukocyte count assay for thrombogenicity screening of biomaterials and medical devices: Evaluation and improvement of ASTM F2888 test standard.

An appropriate preclinical thrombogenicity evaluation of a blood-contacting device is important to reduce thrombosis and thromboembolism risks to patients. The in vitro platelet and leukocyte count assay, as described in the ASTM F2888 test standard, aims to assess thrombogenic potentials of blood-contacting materials. The goals of this study were to evaluate whether this standardized test method can effectively differentiate materials with different thrombogenic potentials and to investigate the impact of anticoagulation conditions on test sensitivity. Using human blood with various anticoagulation conditions, we performed the platelet and leukocyte count assays on four biomaterials and three positive control materials. We found that the use of sodium citrate anticoagulation as stipulated in the 2013 version of the ASTM F2888 standard cannot differentiate materials with different thrombogenic potentials. The modification to use low-concentration heparin, either with recalcified citrated blood or with direct heparinization, substantially improved the test sensitivity and enabled the assay to distinguish platelet count reduction between the positive controls and commonly used biomaterials. Leukocyte count was shown to be a much less sensitive indicator than platelet count for thrombogenicity evaluations of biomaterials. The findings from this study have been incorporated in the recent 2019 version of the ASTM F2888 standard.

Impact of various reagents during blood collection from poultry to analyze morpho-biochemical indicators

The work provides experimental data on the impact of anticoagulants (K3 - EDTA, 3.8% sodium citrate and lithium heparin) and coagulation activator on morpho-biochemical indicators in the blood samples of broiler chickens of cross-selection of Agricultural Center “Smena” at the age of 35 days. The studies were carried out with the use of semi-automatic flow analyzer Sinnowa BS-3000P (SINNOWA Medical Science &amp; Technology Co., Ltd, China) and veterinary automatic blood analyzer DF-50 by Dymind Biotech (PRC) with the use of original reagents. The results showed that the most optimal reagent for determination of morphological indicators of the poultry blood was EDTA. The average white blood count (WBC) is lower in blood samples with citrate and heparin in contrast to EDTA by 30.5% and 24.1%, respectively. The average red blood count (RBC) is lower in tubes with sodium citrate (by 37.9 %) and lithium heparin (by 13.8 %) in contrast to K3-EDTA. When the sodium citrate anticoagulant is used, the blood biochemistry results for all positions of the experiment, excluding calcium, are lower than in heparin, and its closest values are lower than in coagulation activator, excluding trypsin activity. That’s why, the most optimal reagents for biochemistry, including for blood enzyme tests, should be considered the tubes with heparin or coagulation activator.

Treatment Effect of Regional Sodium Citrate Anticoagulation in Elderly Patients With High-Risk Bleeding Receiving Continuous Renal Replacement Therapy.

ObjectiveTo investigate the safety and efficacy of regional citrate anticoagulation (RCA) on elderly patients at high risk of bleeding after continuous renal replacement therapy (CRRT).MethodsA total of 31 patients at high risk of bleeding who received CRRT in the intensive care unit were collected. The patients were divided into RCA group (n = 17) and no anticoagulation group (NA, n = 14) according to whether RCA was used or not. The levels of creatinine (Cr), blood urea nitrogen (BUN), prothrombin time (PT), activated partial thromboplastin time (APTT), total calcium (tCa), ionized calcium ion (iCa2+), sodium ion (Na+), bicarbonate ion (HCO3−), tCa/iCa2+ ratio, and pH were observed after treatment. The filter use time, number of filters used, filter obstruction events, clinical outcomes, and safety evaluation indexes were compared post-treatment.ResultsAfter treatment, serum Cr and BUN levels, APTT and PT levels in the RCA group were significantly lower than the NA group. The tCa, iCa2+, HCO3−, tCa/iCa2+, and pH were within the normal range after RCA treatment while Na+ levels saw a significant increase. In the RCA group, the filter using time was significantly longer, with significantly reduced numbers of filter use within 72 h and filter disorder events. Additionally, patients in the RCA group showed significant recovery of renal function and a significant reduction in bleeding events and in-hospital mortality.ConclusionRCA treatment significantly improves clinical outcome of patients at high risk of bleeding after CRRT, safely and effectively prolongs the filter life and avoids coagulation incidences.

Perbedaan Jumlah Trombosit Platelet Rich Plasma Yang Menggunakan Tabung Natrium Sitrat Dan Tabung ACD-A

Background: PRP is a relatively new field and there is still minor researches that are usage. However, PRP can be applied to a lot of various fields. This study willmake a comparison between tubes that have sodium citrate anticoagulant and tubes that have ACD-A anticoagulants, to be able to find out which of the two are able to produce a higher number of PRPs. Objective: To obtain the anticoagulants that produce more platelet countsand to know the difference in the platelet counts in PRPs found in tubes that have sodium citrate anticoagulants and tubes that have ACD-A anticoagulants. Method: This study was an experimental laboratorium research with an equivalent control group design. Data is normally distributed, so that the parametic statistical analysis test uses Paired T-Test. Results: From 32 respondents, there was an increase in both anticoagulant tubes, where the average increase in platelet counts in PRP using sodium citrate tubes was 337593.75 / μl and the standard deviation was 158795.437 whereas, the average increase in platelet counts in PRP using ACD-A tubes was 909062.50 / μL and the standard deviation is. 284336.005. The difference in platelet counts in PRP using sodium citrate tubes and ACD-A tubes was stated to be significant because the result of the Paired T-Test analysis showed Sig (2-Tailed) = 0,000 (p &lt;0.05). Conclusion: There was a significant difference platelet counts in PRP using sodium citrate tubes and ACD-A tubes. Tubes that have ACD-A anticoagulants had higher results than the ones that have sodium citrate anticoagulant.