Evaluation of platelet concentrates prepared using different methods after overnight holding (18-24 h) of whole blood at room temperature.

BACKGROUND AND OBJECTIVES: Regulatory requirement of fixed holding time (6 h) of whole blood (WB) at room temperature, that is, 22-24°C (RT) results in sub-optimal component separation. The aim was to evaluate the platelet concentrates (PC) prepared by both platelet rich plasma (PRP) and buffy coat (BC) methods after overnight hold (18-24 h) at RT. MATERIALS AND METHODS: A prospective experimental study was performed. A total of 48 WB units collected were divided into four groups (12 each) control-1 (C1) and test-1 (T1) for PRP and control-2 (C2) and test-2 (T2) for the BC method. Control groups were processed within 6 h, and in test groups, components were prepared after overnight hold, followed by evaluation of quality parameters. RESULTS: Irrespective of the method used, all PCs had similar volume, platelet yield, swirling, no bacterial contamination, RBC contamination, PaO2 and PaCO2 levels. PCs in the T1 group had significant differences in glucose and MPV values on d1, which were resolved by d5 of storage. PCs in T2 has significant differences in pH, glucose, and MPV levels throughout storage. PRBC in test and control groups had similar quality parameters till d42 of storage. FFPs in all tests were noninferior to the concurrent control groups till 3 months of storage. CONCLUSION: Overnight holding of WB had no lasting deleterious changes. Though a few biochemical parameters in the test groups were significantly different, they can be accepted to improve the logistics of component separation. Overall PRP method seemed to have a better result than the BC method after an overnight hold.

Real-world evidence of heparin and citrate use in extracorporeal photopheresis: A hypothesis-generating data review of device settings and performance.

Extracorporeal photopheresis (ECP) is widely used for the treatment of cutaneous T-cell lymphoma, graft-vs-host disease, and other immune-related conditions. To avoid clotting during treatment, the ECP system used must be effectively primed with an anticoagulant. Heparin is the recommended anticoagulant for the THERAKOS CELLEX System, but acid citrate dextrose-A (ACDA) is often used. We compared system performance between these two anticoagulants for this ECP system. Deidentified data for ECP device performance were obtained at each treatment session, from automatically logged Smart Cards or labels completed by device operators. We compared the effects of ACDA or heparin on overall treatment duration, buffy coat (leukocyte) collection time, photoactivation time and the number of alarms and warnings. The variability in these parameters was also assessed. Data from 23 334 treat sessions were analyzed; ACDA was used in 34.4% and heparin in 65.6%. Overall, the ECP procedure duration, buffy coat collection time and photoactivation time were numerically similar regardless of whether ACDA or heparin was used, and regardless of needle mode. Photoactivation time variability was lower with ACDA compared with heparin in all needle modes. Among treatments that were completed automatically without any operator intervention, total treatment duration and photoactivation time were significantly reduced with ACDA use in both the double- and single-needle modes. The data presented indicate that, in both double- and single-needle modes, the THERAKOS® CELLEX® integrated ECP system performed similarly with ACDA compared to heparin, although ACDA demonstrated potential benefits in reducing variability in photoactivation time.

Prevalence of bovine trypanosomosis and tsetse fly density in Loka Abaya and Derara districts in Sidama Regional State, Ethiopia.

Animal trypanosomosis is a significant livestock disease with economic and social repercussions, reducing the supply of animal products and restricting the utilization of animals for traction and transportation. In Ethiopia, it is prevalent and poses a major hindrance to the advancement of animal production. This repeated cross-sectional study was aimed at assessing seasonal variation in bovine trypanosomosis prevalence and tsetse fly density and identifying the potential risk factors in the Loka Abaya and Derara districts of the Sidama National Regional State. Blood samples were collected from 964 cattle, 484 samples during the dry season, and 480 during the wet season. The buffy coat method was employed to analyze these samples. Furthermore, 78 standard NGU traps were set up at various locations in the two districts during both seasons for entomological investigation. The overall apparent prevalence of trypanosomosis was 9% (95% CI 7.3-11.0), without a significant difference (p > 0.05) between the dry season (7.4%) and wet season (10.6%). The apparent prevalence was significantly higher in Loka Abaya (11.8%) than in Derara (6.3%) district (OR = 2.04; p = 0.003) and in cattle with black coat color (29%) than in mixed color (6.8%) (OR = 5.3; p < 0.001). The majority of infections were caused by Trypanosoma congolense (70%), followed by T. vivax (29%), and mixed infections (1%) with the two species. The average packed cell volume (PCV) was significantly (p < 0.0001) lower in infected animals (20.7 ± 4%) compared to uninfected ones (25.5 ± 5.4%), in cattle examined during the dry season (24.1 ± 6%) versus the wet season (26.1 ± 4.7%), in cattle sampled from the Loka Abaya district (24.2 ± 5.5%) versus Derara district (26 ± 5.3%), and in cattle with poor body condition (23.6 ± 5.7%) compared to those with good body condition (26.5 ± 5.3%). A total of 5282 flies were captured during the study, with 4437 (84%) being tsetse flies (Glossina pallidipes), 439 (8.3%) Tabanids, 190 (3.6%) Stomoxys spp., and 216 (4.1%) Musca spp. The apparent density (AD) of G. pallidipes was 28.4 flies/trap/day, showing no statistically significant difference between wet (32.1) and dry (24.6) seasons (p > 0.05). The AD of G. pallidipes was significantly (p < 0.001) higher in the Loka Abaya district (57.3) than in the Derara district (0.9). The study highlights a moderate trypanosomosis apparent prevalence and high AD of G. pallidipes, showing significant variation between the study districts but no seasonal difference. The observed apparent prevalence of trypanosomosis and tsetse fly density notably affects animal health and productivity. As a result, strategies for vector control like insecticide-treated targets, trypanocidal medications for infected animals, and community-based initiatives such as education and participation in control programs are recommended.

Growth and Distribution of Bacteria in Contaminated Whole Blood and Derived Blood Components.

Introduction: Bacterial contamination of blood products presumably occurs mainly during blood collection, starting from low initial concentrations of 10-100 colony-forming units (CFUs) per bag. As little is known about bacterial growth behavior and distribution in stored whole blood (WB) and WB-derived blood products, this study aims to provide data on this subject. Methods: WB units were inoculated with transfusion-relevant bacterial species (Acinetobacter baumannii, Bacillus cereus, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Pseudomonas fluorescens, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus dysgalactiae, Streptococcus pyogenes, Yersinia enterocolitica; n = 12 for each species), stored for 22-24 h at room temperature, and then centrifuged for separation into plasma, red blood cells (RBCs), and buffy coats (BCs). The latter were pooled with 3 random donor BCs and one unit of PAS-E each to yield plasma-reduced platelet concentrates (PCs). Samples for bacterial colony counting were collected after WB storage and immediately after blood component production. Sterility testing in PCs (n = 12 for each species) was performed by bacterial culture after 7 days of storage. Results: Bacterial growth in WB varied remarkably between donations and species. Streptococcus species produced the highest titers in WB, whereas Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas fluorescens did not multiply. Centrifugation resulted in preferential accumulation of bacteria in BCs, with titers of up to 3.5 × 103 CFU/mL in BCs and up to ≤0.9 × 103 CFU/mL in BC-derived PCs. Overall, 72/144 PCs (50%) tested positive for bacteria after storage. Sterility test results were species-dependent, ranging from 12 of 12 PCs tested positive for Streptococcus pyogenes to 1 of 12 PCs positive for Escherichia coli. Bacterial contamination of RBC and plasma units was much less common and was associated with higher initial bacterial counts in the parent WB units. Conclusions: Bacterial growth in WB is species-dependent and varies greatly between donations. Preferential accumulation of bacteria in BCs during manufacturing is a critical determinant of the contamination risk of BC-derived pooled PCs.

Increasing the collection flow rate to 2 mL/min is effective and reduces the procedure time in off-line photopheresis.

BACKGROUND: Extracorporeal photopheresis (ECP) treatment, mostly based on apheresis technology, is used for immunomodulation in various diseases such as cutaneous T-cell lymphoma, graft versus host disease and other (auto)immune disorders. The aim of this study was to collect high cell counts and purity in shorter procedure times using an ECP off-line system with an increased collection flow rate of 2 mL/min to a target volume of 200 mL buffy coat. STUDY DESIGN AND METHODS: In this prospective study, data of routinely performed off-line photopheresis treatments were collected and analyzed at the Central Institute for Blood Transfusion & Department of Immunology (ZIB) of the Tirol Kliniken, to assess absolute cell counts and procedure times and to calculate collection efficiencies (CE2). RESULTS: A total of 22 patients participated in this study. The processed blood volume was 4312 mL, the collection time 120 min, overall procedure time 157 min and the absolute cell counts of treated white blood cells (WBC) and mononuclear cells (MNC) were 5.0 and 4.3 × 109 respectively (median values). The calculated CE2 for WBC and MNC was 21.1% and 58.5%, the proportion of treated MNCs of the total number of MNCs present was 55.0%. CONCLUSION: The data presented in this study show high therapeutically effective cell counts collected with a high MNC purity within a shorter overall collection/procedure time due to an increased collection flow rate.

Dose-dependent effects of acetaminophen and ibuprofen on mitochondrial respiration of human platelets.

Acetaminophen and ibuprofen are widely used over-the-counter medications to reduce fever, pain, and inflammation. Although both drugs are safe in therapeutic concentrations, self-medication is practiced by millions of aged patients with comorbidities that decrease drug metabolism and/or excretion, thus raising the risk of overdosage. Mitochondrial dysfunction has emerged as an important pathomechanism underlying the organ toxicity of both drugs. Assessment of mitochondrial oxygen consumption in peripheral blood cells is a novel research field Cu several applications, including characterization of drug toxicity. The present study, conducted in human platelets isolated from blood donor-derived buffy coat, was aimed at assessing the acute, concentration-dependent effects of each drug on mitochondrial respiration. Using the high-resolution respirometry technique, a concentration-dependent decrease of oxygen consumption in both intact and permeabilized platelets was found for either drug, mainly by inhibiting complex I-supported active respiration. Moreover, ibuprofen significantly decreased the maximal capacity of the electron transport system already from the lowest concentration. In conclusion, platelets from healthy donors represents a population of cells easily available, which can be routinely used in studies assessing mitochondrial drug toxicity. Whether these results can be recapitulated in patients treated with these medications is worth further investigation as potential peripheral biomarker of drug overdose.

Quantitative increases of extracellular vesicles in prolonged cold storage of platelets increases the potential to enhance fibrin clot formation.

BACKGROUND: Platelet derived extracellular vesicles (EVs) display a pro-coagulant phenotype and are generated throughout platelet concentrate (PC) storage. Cold storage (CS) of PCs is thought to provide a superior haemostatic advantage over room temperature (RT) storage and could prolong the storage time. However, the effect of storage conditions on EV generation and PC function is unknown. We investigated EV production under CS and RT conditions and assessed whether these EVs exhibited a more pro-coagulant phenotype in model experiments. MATERIALS AND METHODS: Buffy-coat-derived PCs in a platelet additive solution (PAS) to plasma ratio of approximately 65:35 were stored at RT (22 ± 2°C) or CS (4 ± 2°C) for a prolonged storage duration of 20 days. Impedance aggregometry assessed platelet function. EVs were isolated throughout storage and quantified using nanoparticle tracking analysis. EVs were applied to a coagulation assay to assess the impact on fibrin clot formation and lysis. RESULTS: CS produced significantly larger EVs from day 4 onwards. EV concentration was significantly increased in CS compared to RT from day 15. EVs, regardless of storage, significantly reduced time to clot formation and maximum optical density measured compared to the no EV control. Clot formation was proportionate to the number of EV applied but was not statistically different across storage conditions when corrected for EV number. CONCLUSION: EVs in CS and RT units showed similar clot formation capacity. However, the higher number of larger EVs generated in CS compared to RT suggests PC units derived from CS conditions may overall exhibit a haemostatically superior capacity compared to RT storage.

Measurement of yield and quality of DNA in human buffy coat is extraction method dependent.

During the last few years, an important element in the improvement of the molecular biology techniques has been the necessity for availability of high quality and functionality DNA. Several DNA extraction procedures with different results in both performance and quality, have been proposed. In this study our objective was to determine the most reliable extraction method that balances DNA quantity, and to assess the sample quantification of the fluorometric DNA quantification methods. For this, blood extracted by venopunction from 20 healthy volunteers was used to obtain DNA from buffy coat, and 4 commercial DNA extraction kits were assessed as well as two fluorometric DNA quantification methods with protocols of different complexity. Results suggest that manual methods achieve higher quality and larger yields of DNA. DNA purity obtained with the 4 extraction kits evaluated through the 260/280 and 260/230 ratio showed that the Qiacube kit fulfilled the criteria established in this work, followed very close by the Flexigene kit. On the other hand, the fluorometric DNA methods used in the samples quantification showed a higher variability when using QuantiFlour method, obtaining better results probably due to the simplicity of this protocol.

Understanding Solid-Based Platelet-Rich Fibrin Matrices in Oral and Maxillofacial Surgery: An Integrative Review of the Critical Protocol Factors and Their Influence on the Final Product.

Platelet-rich fibrin (PRF) is a second-generation platelet concentrate whose use in clinical practice has been widely disseminated. This has led to the development of several commercial protocols, creating great confusion as to the terminology and implications of each of them. This integrative review aims to identify the critical factors of each of the phases of the solid-based PRF matrix protocol and their possible influence on their macro- and microscopic characteristics. An electronic search of the MEDLINE database (via PubMed), Web of Science, Scopus, LILACS, and OpenGrey was carried out. The search was temporarily restricted from 2001 to 2022. After searching, 43 studies were included that met the established criteria. There were numerous factors to consider in the PRF protocol, such as the material of the blood collection tubes, the duration of phlebotomy, the parameters related to blood centrifugation, the time from centrifugation to dehydration of the fibrin clots and their dehydration into membranes, as well as the time to clinical use. These factors influenced the macro- and microscopic characteristics of the PRF and its physical properties, so knowledge of these factors allows for the production of optimised PRF by combining the protocols and materials.

The history of buffy coat platelet concentrates: The Dutch story.

The buffy coat method as a source for platelet concentrates was developed in the 1970s and is still used in many blood centres around the world. Development of the method sparked various technological advances in blood collection, processing and storage. At the time, the need for platelet concentrates sharply increased because of better treatment regimens for (onco)haematological diseases, which forced blood centres to standardize and automate their production processes as much as the technology would allow. In this review, a historical overview of the Dutch experiences is provided in the context of the international developments.

Evaluation of platelet concentrates prepared from whole blood donations with collection times between 12 and 15 min: The BEST Collaborative study.

BACKGROUND AND OBJECTIVES: In many countries, whole blood (WB) donations with collection times between 12 and 15 min are not allowed to be used for platelet concentrates (PC). Since the development of guidelines, many process-related changes have been introduced. We aimed to determine the effect of WB with long collection times on PC quality. MATERIALS AND METHODS: Five participating centres tested buffy coat (BC)-derived PC in platelet additive solution type E prepared from only WB collections lasting <12 min (control) versus similar PC including one BC from a collection lasting >12 min (study group, n = 8). One centre produced platelet-rich plasma (PRP)-derived PC from single donations (<10 or >12 min). All PC were stored at 22 ± 2°C and sampled on Days 1, 6 and 8 post-collection for in vitro quality determination. RESULTS: Average collection time was significantly longer in the study group compared to controls (8.9 ± 2.6 vs. 7.3 ± 1.3 min, p < 0.001). There were no differences in volume, platelet concentration, basal CD62P expression, soluble-CD62P and CCL5 levels, or nucleotide content between the groups. Stimulation with TRAP-6 resulted in comparable levels of cell surface CD62P. On Day 8, all PC fulfilled requirements for pH. The findings from single PRP-derived PC centre were similar. CONCLUSION: PC with one BC and single PRP derived from collections lasting >12 min had equivalent in vitro quality to controls during storage. This study provides evidence that 12-15 min donations should not be excluded for PC preparation and justifies to readdress the guidelines to <15 min instead of <12 min of collection in line with current practice in some countries.

Optimized simple and affordable procedure for differentiation of monocyte-derived dendritic cells from LRF: An accessible and valid alternative biological source.

Dendritic cells are one of the most popular immune cells, which plays a remarkable role in both immunotherapy and tolerance induction. Due to unwanted side effects of leukocyte presence in donated blood, the policy of blood service is the pre-storage reduction of leukocytes, which today, filtration is the most common method for this purpose. The filtration method has led to diminished access to Buffy coat as a generally used conventional source of biological cells. We developed a simple, affordable, and reproducible method for dendritic cell differentiation from filter-derived monocytes and, the results of the filter study were compared with differentiated DCs from the conventional buffy coat-derived monocytes. The Monocytes were recovered from leukoreduction filter using an optimized protocol with supplemented PBS buffer. Following the adhesion method, CD14+ Monocyte-enriched population with the purity of 94 % was obtained. After cytokine stimulation over a 6-day period and maturation induction by LPS, differentiated DCs were evaluated for morphology, surface markers (CD86, CD40, CD83 and, HLA-DR), antigen uptake potency and IL-12 secretion. Analysis and comparison of the results represented no significant difference between the two groups. Accordingly, we conclude that leukoreduction filters could be introduced as a reliable and research-grade source of monocyte for DC generation in biological research.

Buffy Coat Transcriptomic Analysis Reveals Alterations in Host Cell Protein Synthesis and Cell Cycle in Severe COVID-19 Patients.

Transcriptome studies have reported the dysregulation of cell cycle-related genes and the global inhibition of host mRNA translation in COVID-19 cases. However, the key genes and cellular mechanisms that are most affected by the severe outcome of this disease remain unclear. For this work, the RNA-seq approach was used to study the differential expression in buffy coat cells of two groups of people infected with SARS-CoV-2: (a) Mild, with mild symptoms; and (b) SARS (Severe Acute Respiratory Syndrome), who were admitted to the intensive care unit with the severe COVID-19 outcome. Transcriptomic analysis revealed 1009 up-regulated and 501 down-regulated genes in the SARS group, with 10% of both being composed of long non-coding RNA. Ribosome and cell cycle pathways were enriched among down-regulated genes. The most connected proteins among the differentially expressed genes involved transport dysregulation, proteasome degradation, interferon response, cytokinesis failure, and host translation inhibition. Furthermore, interactome analysis showed Fibrillarin to be one of the key genes affected by SARS-CoV-2. This protein interacts directly with the N protein and long non-coding RNAs affecting transcription, translation, and ribosomal processes. This work reveals a group of dysregulated processes, including translation and cell cycle, as key pathways altered in severe COVID-19 outcomes.

Fibrinogen Concentrations in Liquid PRF Using Various Centrifugation Protocols.

Liquid platelet-rich fibrin (PRF) is produced by fractionation of blood without additives that initiate coagulation. Even though liquid PRF is frequently utilized as a natural source of fibrinogen to prepare sticky bone, the concentration of fibrinogen and the overall amount of "clottable PRF" components have not been evaluated. To this aim, we prepared liquid PRF at 300, 700, and 2000 relative centrifugal force (RCF), for 8 min and quantified the fibrinogen levels by immunoassay. We report here that, independent of the RCF, the fibrinogen concentration is higher in the platelet-poor plasma (PPP) compared to the buffy coat (BC) fraction of liquid PRF and further decreases in the remaining red fraction. We then determined the weight of the clotted PRF fractions before and after removing the serum. The PPP and BC fractions consist of 10.2% and 25.3% clottable matrix suggesting that more than half of the weight of clottable BC is caused by cellular components. Our data provide insights into the distribution of fibrinogen in the different fractions of liquid PRF. These findings suggest that PPP is the main source of clottable fibrinogen, while the BC is more a cell source when it comes to the preparation of sticky bone.

Laboratory diagnosis of community-acquired bloodstream infection in therapeutic pathology.

Community-acquired bloodstream infections (CBSIs) occur in the out-of-hospital setting (44%) and increase the overall mortality from bloodstream infections (BSIs) by 7.2% per year. The development of CBSIs depends on both comorbid and polymorbid diseases and the patients' age. The causes of CBSIs are: respiratory, hepatobiliary gastrointestinal and urogenital tracts and dental interventions. The etiology of CBSIs is characterized by the isolation of coagulase-negative staphylococci (CNS) (32%), E. coli (27%). To investigate community-acquired bloodstream infection in therapeutic patients. The study included out-of-hospital patients (n=382). 4.5 ml of blood were taken intravenously into a closed vacuum system in order to obtain a buffy coat of blood, which was put on glasses for microscopy and Petri dishes with blood agar for cultivating under aerobic and anaerobic conditions. Microorganisms were identified by mass spectrometry. Microscopy of blood smears was used for rapid diagnosis of infection in the bloodstream. BSI was diagnosed in 183 (48.0%) out of 382 out-of-hospital patients. The etiology of CBSIs was studied on 297 isolated strains of microorganisms. CBSIs rather often complicated the underlying disease in women and young people. The spectrum of CBSI pathogens included aerobic and anaerobic bacteria and fungi. Gram-positive cocci with the leadership of S.epidermidis (25.7%) were more often isolated among bacteria. 70% of all isolated pathogens grew under anaerobic conditions. CBSIs were characterized by polymicrobiality (33.5%) of two to four different microorganisms in one blood culture; the species of associates of polymicrobial blood cultures are shown. Microscopic examination of blood smears revealed microorganisms in 97.1% of cases, including associations of bacteria with fungi (66.9%). CBSIs occurred after contour plastic, in diseases of the respiratory system, genitourinary system, oral cavity, skin and subcutaneous tissue. Microbiological examination of the buffy coat is an alternative microbiological method of CBSIs diagnosis, which includes microscopy and blood cultivating and has a high diagnostic efficiency (97.1% and 48% respectively). It can become an option for replacing imported blood culture automated systems.

Comparison of capillary, venous and buffy coat blood samples in detecting Plasmodium species among malaria suspected patients attending at Hamusite health center. A cross-sectional study.

BACKGROUND: Both capillary and venous blood samples have been interchangeably used for the diagnosis of malaria in Ethiopia. However, Plasmodium parasites are thought to be more concentrated in capillary than in venous blood. Hence, selecting a sample source where parasites are more concentrated is indispensable approach in order to maximize the accuracy of blood film microscopy. Therefore, the present study aimed to compare the detection rate and the parasitemia level of Plasmodium species from conventional capillary and venous blood films, and buffy coat preparations. METHODS: A facility based cross-sectional study was conducted from Feburary to March 2020 among 210 febrile patients attending Hamusite health center, northwest Ethiopia. Capillary and venous blood samples were collected and buffy coat was prepared from each sample. Thin and thick blood films were prepared, stained, and examined microscopically following standard protocol. Data were analysed using Statistical Package for Social Sciences Software version 20 and Med-Calc software version 19.3. RESULTS: Capillary blood buffy coat (61/210, 29.0%) had significantly higher detection rate as compared to capillary (48/210, 22.9%) and venous (42/210, 20.0%) blood films (p < 0.001). However, no significant difference was observed between capillary and venous blood films (p = 0.070) in detecting Plasmodium species. The highest and the lowest mean asexual stage parasite counts were found in capillary blood buffy coat (4692.88) and venous blood (631.43) films, respectively showing significant variations (p < 0.001). Mean gametocyte count was also highest in capillary blood buffy coat (3958.44). As compared to capillary blood buffy coat, the sensitivity of venous blood buffy coat, capillary blood film and venous blood film were 73.8, 78.7, 68.9%, respectively. CONCLUSION: Capillary blood buffy coat samples showed the highest sensitivity in detecting and quantitating malaria parasites that its use should be promoted in clinical settings. However, conventional capillary and venous blood films could be used interchangeably.

Effect of two different parts of CGF on post-extractive alveolar ridge preservation: a preliminary histomorphometric analysis in a Split-Mouth design.

Tooth extraction produces alveolar bone resorption and soft tissue remodelling, so identification of adequate technique for alveolar ridge preservation after tooth extraction is fundamental for all specific cases. Among the several biomaterials, CGF can represent an ideal alternative considering its and its mechanical and biological properties. In this preliminary study we compared the effectiveness of the use of two different parts of CGF (WP-White Part and BC-Buffy Coat) versus natural healing (CTR) by a split-mouth randomized clinical design. Four healthy patients who needed extraction of three teeth were selected. Post-extractive alveolar sockets were filled randomly with CGF-WP, CGF-BC or nothing for CTR. After 60 days, before implant placement, a biopsy for each alveola was obtained for quantitative histomorphometric analysis. The data obtained showed that the use of CGF-WP could achieve good regenerative results, supporting the use of this part for the preservation of the post-extractive alveolar socket.

A comparative analysis of factors influencing haemoglobin content in RBC units.

BACKGROUND AND OBJECTIVES: The increment in a patient's haemoglobin level is based upon the haemoglobin content of the transfused RBC units. The total haemoglobin present in the blood bags can vary because of the blood donor, processing method, volume and type of bag used. The study is done to analyse the factors causing variation of haemoglobin content in RBC units. MATERIALS AND METHODS: A total of 260 RBC units were tested for the haemoglobin content and analysed with the donor variables (age, gender, weight & capillary haemoglobin). The blood bags were then separated into two groups based on the donor capillary haemoglobin (normal 12.5-15.0 g/dL vs high 15.1-18.0 g/dL), volume (350 vs 450 mL), processing method (Platelet rich plasma vs buffy coat) and further analysis was carried out. RESULTS: The mean haemoglobin content was 54.7 g ranging from 34.2-80 g per unit. The factors which significantly influenced (p < 0.0001) were capillary haemoglobin, gender and weight of donor, volume of blood collected and the processing method. There was a significant difference (p < 0.0001) in haemoglobin content between the two groups in all the three categories (capillary haemoglobin, volume and processing method). Regression analysis showed all three of them contributed to 80 % variability of haemoglobin content in the RBC unit. CONCLUSION: The marked variation of haemoglobin content in our study revealed that there is a need for standardizing RBC unit. Labelling of units with haemoglobin content and transfusion based on it will result in better patient care.

Donor platelet and leukocyte count as predictive factors of the quality of platelet concentrates obtained from whole blood by semiautomated fractionation.

Platelet concentrates (PCs) obtained from whole blood are produced by fractionation of the buffy coat (BC) or the platelet-rich plasma. Despite the improvements in the technologies used for the hemocomponent fractionation, the proportion of PCs that do not accomplish the quality requirements is high. This study aimed to determine whether the basal platelet and leukocyte counts are predictive factors of the quality of the PCs obtained from BC by semiautomated fractionation. Quality control registers of 196 PCs were analyzed. Gender- and age-dependence of the blood cell count and the characteristics of PCs were evaluated. Platelet yield and residual leukocytes in the PCs were correlated with the platelet and leukocyte counts and the age of the donors. Predictive efficacy was assessed, and an optimal cut-off was established. The proportions of PCs accepted and rejected by using or not the optimal cut-off were compared. 50.0% of the PCs accomplished all the quality control requirements. Female donors had a higher basal platelet count than males. A correlation was observed between basal platelets and platelet yield, but not between basal leukocytes and residual leukocytes. The basal platelet count predicted the quality of the PCs. A cut-off of 231,000 platelets/mm3 was established, but it did not improve the proportion of accepted PCs. In conclusion, we found that the basal platelet count is correlated with the platelet yield. The basal leukocyte count is not correlated with the residual leukocytes. The established cut-off for the basal platelet count did not improve the proportion of accepted PCs.

Comparison of in-vitro and in-vivo parameters of whole blood derived random donor platelets (RDP) after over-night hold with RDP prepared after 2-h hold: Single centre report from India.

BACKGROUND: Random Donor Platelet (RDP) derived from whole blood is the major source of platelets in India. At our centre, we prepare RDPs by buffy coat method after a holding period of 2-hours (THRDP) as per current regulatory guidelines. Overnight hold of buffy coats before RDP preparation (OHRDP) would logistically optimise the manpower usage at our centre. The aim of this study was to compare both in-vitro as well as in-vivo parameters of OHRDPs with THRDPs. METHODOLOGY: Hematological (Platelet, leucocyte counts), physical (pH and Swirling) and biochemical parameters (pO2, pCO2, lactate, bicarbonate and glucose) as well as platelet activation markers were tested in THRDPs and OHRDPs each at Day-1 and Day-5 as in-vitro studies. Separately, in-vivo study was done where Corrected count increment (CCI) and percentage platelet recovery (PPR) were considered. All parameters were expressed as Mean ± Standard deviation and were analysed using paired t-test with level of significance, p < 0.05. RESULTS: OHRDPs had higher platelet counts and lower leucocytes and CD62 P expression than THRDPs. All other markers were well within the quality control range in both groups. No significant differences were seen in the two groups when comparing CCI and PPR. CONCLUSION: OHRDPs were found to be as good or better as compared to the THRDPs in the in-vitro part of our study. Additionally, there were no significant differences between the two groups when they were compared in vivo. This makes us conclude that overnight hold of buffy coats may be implemented at our center.