Local Blood Bank Research Articles

Development and Implementation of a Standard Operating Procedure for Military Working Dog Blood Collection, Storage, and Transport.

Military working canines are critical assets and force multipliers for the Joint Force. Most often deployed forward of Role 2 assets, they are reliant on non-veterinary resources when wounded, ill, or injured in an operational environment. Hemorrhagic shock is the most prevalent form of shock seen in battlefield injuries and is most effectively treated with whole blood transfusion. Dogs cannot be transfused with human blood and there is no formal Department of Defense (DoD) canine blood product distribution system to operational settings. A walking blood bank is helpful when multiple dogs are geographically co-located and the resource can be provided to an injured patient quickly. In areas as widely dispersed as the Horn of Africa, the likelihood of co-location is slim and delaying this vital resource can mean the difference between life and death. Therefore, personnel at the Role 2 facility in Camp Lemonnier, Djibouti, filled a critical capability gap for the operational area by producing a local canine whole blood bank with distribution to multiple countries. This protocol can be replicated by other locations to improve medical readiness for the working canines who serve to maintain DoD Force Protection.

Evaluation of Two Osmosis-Based Methods for the Preparation of Drug Delivery Systems Based on Red Blood Cells.

Erythrocytes have been thoroughly investigated as drug delivery systems for a wide range of therapeutic molecules and using different kinds of loading methods, outstanding the osmosis-based methods as the most used ones. Most of them involve too much handling of blood components and the immediate obtention of fresh blood. Based on our group's considerable experience in dialysis-based carrier erythrocyte preparation, this study details a simple method based on hypotonic dilution and subsequent resealing that has been developed for stavudine using packed erythrocytes from a local blood bank. Properties of the obtained carrier erythrocytes were studied in comparison to those prepared by dialysis. Erythrocytes' morphology, osmotic fragility, hematological parameters, and in vitro release profiles were evaluated. Loaded erythrocytes obtained with the proposed method did not show impaired properties in comparison with those obtained with our reference method, provided that the buffer composition remained the same. In the present work, we have optimized a simplified method for erythrocytes' drug loading, which can use blood transfusion products and could be easily automatized and scalable.

Importance of selected ABCB1 SNPs for the level of severity of depressive symptoms and effectiveness of recurrent depressive disorder therapy

The expression level of mRNA and also the function of P-gp are strictly connected with the polymorphic nature of the ABCB1 gene. In this study, we evaluated the association between promoter SNP, i.e. T-129C, three other SNPs investigated earlier and ABCB1 expression in the depression group. To assess the additive significance of these SNPs on clinicopathological features a mathematical model was also built.102 patients suffering from recurrent depressive disorder (rDD) and 94 healthy individuals from a local blood bank were enrolled in this study. ABCB1 gene polymorphism was identified by the RFLP method. The relative level of ABCB1 expression was measured by real-time PCR.For SNP T-129C no statistically significant differences in allele and genotype frequencies between depression and control groups were found (p = 0.3176). There was no statistically significant association between the expression value and 4 studied SNPs in ABCB1 (T-129C, C1236T, G2677T/A and C3435T) or the investigated clinicopathological features. Furthermore, a correlation between the initial HDRS score (lower than 23) and presence of at least 1236 T allele was observed, in particular in combination with 3435 T or 2677 T/A. Mutated allele of each SNP was also significantly associated with declined response to antidepressant therapy, both individually and in combination with others.Results of this study suggest that T-129C does not play an important role in the rDD development. The influence of the studied SNPs on ABCB1 gene expression is still unknown. However, the additive impact of 3 most frequently studied SNPs of ABCB1 on the course of depression and effectiveness of its treatment was confirmed.

Cytoskeletal changes in Erythrocytes during storage in banked blood

Introduction: Erythrocytes have flexible, non-nucleated bi-concave shape with lipid bilayer cytoskeleton. Any alterations of erythrocyte shape make it susceptible to hemolysis. Blood for transfusion purpose is routinely stored in Citrate Phosphate Dextrose Adenine (CPDA-1) containing blood bags. During storage, blood undergoes an array of different morphological changes termed as “storage lesions” which makes it more fragile. This study was aimed to determine the structural and functional modifications in erythrocytes in CPDA-1 blood stored in local blood bank of KPK. Material & Methods: Blood from twenty healthy volunteer donors was taken and kept in CPDA-1 containing blood bags at Institute of Basic Medical Sciences (IBMS), Khyber Medical University (KMU). Hb-levels and Erythrocyte, Reticulocyte counts, Mechanical Fragility Index (MFI) and immunofluorescence staining for ankiyrin1 protein were performed on fresh blood samples. Samples for reticulocyte count was taken for 5 consecutive days while for the remaining parameters, blood was taken at 5 days interval till day 20th. The light and fluorescence micrographs were obtained accordingly and osmotic fragility tests were performed. Results: A significant mean reduction in erythrocyte counts and Hb-level was observed from day 0 to 20 (p=0.001), while MFI increased from day 0 to day 20 (12.88%±7.58 p=0.001). Reticulocyte count also decreased from day 0 up to day 5 (p=0.001). A weak association was observed between changes in MFI and erythrocyte morphology from day 0 to 20 (r=0.349), while the intensity and pattern of ankyrin1 protein expression appeared to change from day 10. Conclusion: Blood stored for a week has same properties as fresh blood, however, important structural alterations start to appear after the first week of storage and worsen with time. Therefore, to gain better transfusion results, blood stored for up to one week can safely be transfused.

Perfusion indices can predict early volume depletion in a blood donor model.

Blood donation from healthy donors is used experimental model that surrogates for class 1 hemorrhage in humans. We examined changes in the perfusion index (PI) and plethysmographic variability index (PVI) in healthy blood donors after donating a unit of blood, and we evaluated the usability of these indices in detecting blood loss volumes of less than 750mL (class 1 hemorrhagic shock trauma patients). This study is a prospective, cross-sectional study. 180 healthy volunteers aged 18 and over, who donated blood at the local blood bank, were included in the study consecutively. The age, gender, and body mass index of the volunteers were recorded and, before and after the blood donation, the vital signs and perfusion indices were measured. Of the donors, 61.7% were men (n = 111), and the median age of all donors was 32 (IQR: 21-39). A statistically significant difference was found between the hemodynamic parameters and PIs before and after the blood donation (p < 0.01 for all parameters; median difference of PI [-1.45, 95% CI: (-0.9)-( -2)], median difference of PVI [6, 95% CI: 7.77-4.23]. We evaluated the perfusion indices in the early diagnosis of blood volume loss in patients admitted to the emergency department due to trauma. After the participants donated one unit of blood, we found that their PI decreased and PVI increased compared to the measurements before the blood donation. Considering that major bleeding starts in the very early stage as minor bleeding, it is essential for emergency physicians to recognize class 1 hemorrhagic shock patients. Further, non-invasive and straightforward procedures, such as measuring PI and PVI, can be particularly useful in identifying blood loss volumes of less than 750mL.

AB0077 CONTRIBUTION OF NOTUM TO THE DEVELOPMENT OF OSTEOARTHRITIS

Background:Osteoarthritis (OA) is a degenerative disease characterized by altered homeostasis of joint cartilage and bone, the functionality of which relies on chondrocytes and osteoblasts, that leads to the formation of a defective extracellular matrix (ECM). The ECM plays an essential role in bone biology as it provides the structure of cartilage which serves as a template for bone formation. Collagen X, main component of the ECM, has been described by our group as down-regulated in OA [1]. Our data also points to an important role of the Wnt pathway in OA [1,2]. Furthermore, Wnt proteins have been reported to inhibit chondrogenesis [3], and the Wnt pathway and its modulators have gained attention [4]. Glypicans (GPC1 to GPC6) and NOTUM, among others, have been identified as modulators of this pathway [5,6]. Notably, due to its highly specific inhibition of the Wnt pathway, NOTUM has been proposed as a therapeutic target in conditions with a high activity of the Wnt pathway is involved, such as OA [7].Objectives:We hypothesize that modulators of the Wnt pathway are involved in the development of OA. The aim of this study is to evaluate the presence of Glypicans and NOTUM in the serum of OA patients and healthy individuals in order to determine whether significant differences exist and could clarify their likely involvement in OA.Methods:Peripheral blood samples were obtained from OA patients during routine rheumatologist hospital visits. OA diagnosis was established according to the ACR criteria. Samples from healthy individuals were obtained from the local Blood Bank. In both cases, blood samples were centrifugated (2000g, 15 minutes, 10°C) and serum was obtained.Quantitative ELISA assays for GPC1-6 and NOTUM were carried out using commercial kits (Human GPC1 ELISA Kit, #E-EL-H1710, Elabscience; Human GPC2 ELISA Kit, #E-EL-H1711, Elabscience; Human GPC3 ELISA Kit, #E-EL-H1712, Elabscience; Human GPC4 ELISA Kit, #E-EL-H1713, Elabscience; Human GPC5 ELISA Kit, #ELH-GPC5, RayBiotech; Human GPC6 ELISA Kit, #CSB-EL009708HU, Cusabio; Human Protein NOTUM homolog ELISA Kit, #EK3787, Sab Biotech) and measured in a plate reader (Heales MB-580,Shenzhen Heales Technology Development Co. Ltd.). Protein concentration in serum was calculated using GraphPad Prism 7 software. Differences between samples were analysed with Mann-Withney U test. Significance level set wasp&lt;0.05.Results:Serum from 40 OA patients and 40 healthy donors were included in the study. There were no differences between groups (Table 1).Table 1.Cohort descriptionControl group (n=40)OA group (n=40)Age66,82±5,7569,59±11,24Women (%)32 (80%)30 (75%)Out of 7 proteins analyzed, only NOTUM showed a significant difference between healthy and OA groups (MedianOA=0.4451ng/mL, MedianCONTROL=0.8263ng/mL,p=0.0013). Besides, GPC4 showed an approaching formal significance (MedianOA=0.1254ng/mL, MedianCONTROL=0.1596ng/mL,p=0.0767). The rest of Glypicans analyzed showed no significance differences between groups (GPC1, MedianOA=0.1346ng/mL, MedianCONTROL=0.1190ng/mL,p=0.2379; GPC2, MedianOA=2.593ng/mL, MedianCONTROL=2.955ng/mL,p=0.7489; GPC3, MedianOA=2.024ng/mL, MedianCONTROL=1.422ng/mL,p=0.3574; GPC5, MedianOA=3.663ng/mL, MedianCONTROL=5.529ng/mL,p=0.8829; GPC6, MedianOA=0.3922ng/mL, MedianCONTROL=0.3558ng/mL,p=0.3212).Conclusion:Our results suggest that low levels of NOTUM may contribute to the development of OA. The lack of this inhibitor promotes the activation of the Wnt pathway, high activity of which has been related with OA.

In-vitro performance of a low flow extracorporeal carbon dioxide removal circuit.

Extracorporeal gas exchange requires the passage of oxygen and carbon dioxide (CO2) across an artificial membrane. Current European Union regulations do not require the transfer to be assessed in models using clinically relevant haemoglobin, making it difficult for clinicians to understand the CO2 clearance of a membrane, and how it changes in relation to sweep gas flow through the membrane. The characteristics of membrane CO2 clearance are described using a single membrane at different sweep gas flows in an in vitro model with clinically relevant haemoglobin concentrations using three separate methods of calculating CO2 clearance. To define the CO2 removal characteristics of the extra-corporeal CO2 removal (ECCO2R) device, we devised an in-vitro gas exchange circuit formed by a dedicated ECCO2R circuit (ALung, Pittsburgh, USA) in series with two membrane oxygenators. The system was primed with donated expired human red cells provided by the local blood bank. The experimental set-up allowed constant CO2 input (via one membrane oxygenator) with variable removal from a portion of the blood in a manner which was analogous to that seen in vivo. Blood gases were measured from different ports in the circuit in order to measure the experimental membrane CO2 clearance (VCO2). Results demonstrate that the relationship between VCO2 and gas flow at a constant blood flow of 0.4 L/minute with a haemoglobin of 7 g/dL increases sharply from a gas flow of 0 to 2 L/min but plateaus at gas flows >4 L/minute. VCO2, calculated using three different methods, showed a strong linear correlation with minimal bias. The CO2 clearance of the membrane used in this bench test is non-linear. This has implications for clinical practice, especially during the weaning phase of the device.

PS1594 EXPERIENCE WITH THE NEUTRALIZING AGENT DARAEX® IN BLOOD COMPATIBILITY TESTS IN PATIENTS TREATED WITH DARATUMUMAB

Background:Daratumumab is a human monoclonal antibody used for the treatment of multiple myeloma, which binds to CD38, a protein that is expressed on red blood cells (RBCs). Therefore, the plasma of these patients reacts with the RBCs producing a panreactivity and interfering in the transfusion compatibility testing. Panagglutination may persist for up to 6 months after the last infusion of daratumumab. An anti‐CD38 antibody for treatment of multiple myeloma shows a strong interference in the indirect antiglobulin test (IAT) where most of the reactions turn to unspecifically (wrong) positive. The time consuming standard technique using Dithiothreitol (DTT) counteracts the interference but has major drawbacks like destruction of Kel antigens or hemolysis. The DaraEx® compound inhibits the agglutination effect of anti‐CD38 in IAT without side effects. DaraEx® is a neutralizing agent for the inhibition of the agglutination effect of the anti‐CD38 antibody daratumumab in IAT.Aims:To validate the procedure to resolve the interference of daratumumab in transfusion compatibility testing using red cells treated with DaraEx® for the inhibition of the agglutination effect of the anti‐CD38 antibody daratumumab in IAT. Daratumumab can interfere with crossmatching and antibody screening in the IAT, which will allow us to identify a clinically significant antibody that has been initially masked by the presence of daratumumab.Methods:The study has been conducted in 25 plasma samples of patients diagnosed with multiple myeloma who have been treated with daratumumab. Irregular antibody screening tests and CrossMatching were performed on all of them, being positive in all cases. We performed the technique to eliminate reactivity by treating the red cells used in compatibility tests with DaraEx®, which neutralizes CD38 on the surface of the erythrocyte, thereby preventing Daratumumab from binding and inducing agglutination. The material used was DaraEx®, 0.9% NaCl LISS/Coombs ID‐Cards and Test cell reagents for the ID‐System; We performed the method in Biovue Column Agglutination Technology and gel card systems.Results:After performing the technique, we were able to eliminate panreactivity in both Irregular antibody screening tests and CrossMatching.Summary/Conclusion:Daratumumab causes panreactivity in vitro by binding to CD38 on reagent RBCs. It is necessary to do a baseline antibody screen (type and screen) before starting the treatment with daratumumab. Treating reagent RBCs with DaraEx® is a useful easy and quick method to mitigate the interference created by antiCD38 mAbs in pretransfusion testing. DaraEx is a quick and simple procedure; it doesn’t affect other antigens or alloantibody reactions. It doesn’t have side effects like destruction of blood group antigens or hemolysis as described for the standard DTT treatment. Therefore, we consider it a very useful technique to resolve interferences produced by daratumumb with blood compatibility testing. We believe that it is essential to have a validated technique to resolve these discrepancies. If an emergency transfusion is required, noncrossmatched, ABO/RhD‐compatible RBCs can be given, per local blood bank practices.

Platelet PGD: Effect on Platelet Waste and Financial Efficiency in a Local Blood Bank

<h3>ABSTRACT</h3> Maintaining a blood product supply is essential to optimal patient care; however, daily use is difficult to anticipate. Platelet (PLT) products are the most variable in daily use, have short shelf lives, and are expensive to produce, test, and store. As a result of need, unpredictable demand, and short half-life, PLTs are frequently wasted because of expiration. The purpose of this study is to monitor the use of the Verax Biomedical Platelet Preimplantation Genetic Diagnosis (PGD) Test (PPT) on managing PLT waste and cost. Minimal research has been conducted on PPT’s financial impact. PPT is used for bacterial monitoring and allows for the increased storage of PLTs by up to 7 days. Its implementation should reduce waste and contribute to efficient cost management of on-hand supplies. A secondary analysis was performed to compare data between a 6-month period in 2016 and the same 6-month period in 2017. The audit reviewed the use and waste of PLT apheresis products (aPPT) prior to 2016 and after the implementation of PPT in 2017 at a local level 1 trauma center blood bank. A financial analysis was performed to determine the percent of cost attributed to both use and waste for each timeframe, with the addition of included PPT implementation cost for 2017. With the implementation of PPT, there was an average waste decrease of 43.48% from the 2016 to the 2017 6-month time periods and an estimated $5,300 in savings over the 6 months. With PPT implementation, the blood bank was able to use 93.22% of their cost from the total cost of aPLTs ordered, including the cost of testing in the 2017 time period in comparison to 89.23% of cost use in the 2016 time period. The implementation of PPT was useful in monitoring inventory and extending the on-hand supply of PLTs.

Addressing the Challenges and Issues of Blood Donation via Ubiquitous Computing

Blood is undeniably essential to save lives. The high demand for blood cannot compete with the amount collected through blood donation. In order to cater to the issue, researchers tend to focus more on the donors’ side. Meanwhile, the other party which is the blood bank continues to play its role to increase the blood supply. The blood bank processes blood in four stages: donation, screening, inventory and hospital pickup. This paper aims to address the challenges and issues in the local blood bank through ubiquitous computing, specifically at the blood donation stage. Blood donation is the most critical stage where the blood bank engages directly with the donors. The issues and challenges faced by the blood bank are uncovered by, interviews, field study and literature reviews. The proposed solution takes advantage of the ubiquitous computing concept that enables devices to communicate with each other seamlessly. With the advancement of mobile technology, smartphones become the closest of any devices to achieve ubiquitous computing. Communication technologies like Near Field Communication and Wi-Fi aid the interaction between the user and the system which are expected to solve the challenges and issues of blood donation.

Seroprevalence of IgA anti Epstein-Barr virus is high among family members of nasopharyngeal cancer patients and individuals presenting with chronic complaints in head and neck area.

Epstein-Barr (EBV) infection and presence of a nasopharyngeal cancer (NPC) case in the family increases the risk of developing NPC. Aberrant anti-EBV immunoglobulin A (IgA) antibodies (EBV-IgA) may be present in the sera of non-cancer individuals and predict NPC. Limited studies report the presence of EBV-IgA antibodies within non-cancer individuals in Indonesia where the disease is prevalent. This study aimed at exploring whether EBV-IgA was found more frequently among first degree relatives of NPC patients and individuals presenting with chronic symptoms in the head and neck area compared to healthy controls. A total of 967 non-cancer subjects were recruited, including 509 family members of NPC cases, 196 individuals having chronic complaints in the head and neck region, and 262 healthy donors of the local blood bank. Sera were analyzed using a standardized peptide-based EBV-IgA ELISA. Overall, 61.6% of all individuals had anti-EBV IgA reactivity equal to or below cut off value (CoV). Seroreactivity above CoV was significantly higher in females (38.7%) compared to males (28.7%) (p = 0.001). Older individuals had more seroreactivity above CoV (42.5%) than the younger ones (26.4%) (p< 0.001). Seroprevalence was significantly higher in family members of NPC patients (41.7%), compared to 32.7% of individuals with chronic head and neck problems (p = 0.028) and 16.4% healthy blood donors (p< 0.001). As conclusion, this study showed a significant higher seroprevalence in healthy family members of NPC cases and subjects presenting with chronic symptoms in the head and neck area compared to healthy individuals from the general community. This finding indicates that both groups have elevated risk of developing NPC and may serve as targets for a regional NPC screening program.

Previous Cryopreservation Alters the Natural History of the Red Blood Cell Storage Lesion.

During storage, packed red blood cells (pRBCs) undergo a number of biochemical, metabolic, and morphologic changes, collectively known as the "storage lesion." We aimed to determine the effect of cryopreservation on the red blood cell storage lesion compared with traditional 4°C storage. Previously cryopreserved human pRBCs were compared with age-matched never-frozen pRBCs obtained from the local blood bank. The development of the red cell storage lesion was evaluated after 7, 14, 21, 28, and 42 days of storage at 4°C in AS-3 storage medium. We measured physiological parameters including cell counts, lactic acid, and potassium concentrations as well as signs of eryptosis including loss of phosphatidylserine (PS) asymmetry, microparticle production, and osmotic fragility in hypotonic saline. Compared with controls, previously cryopreserved pRBC at 7 days of storage in AS-3 showed lower red cell counts (3.7 vs. 5.3 × 10 cells/μL, P < 0.01), hemoglobin (Hgb) (12.0 vs. 16.5 g/dL, P < 0.01), hematocrit (33.0% vs. 46.5%, P < 0.01), and pH (6.27 vs. 6.72, P < 0.01). Over 28 days of storage, storage cryopreserved pRBC developed increased cell-free Hgb (0.7 vs. 0.3 g/dL, P < 0.01), greater PS exposure (10.1% vs. 3.3%, P < 0.01), and microparticle production (30,836 vs. 1,802 MP/μL, P < 0.01). Previously cryopreserved cells were also less resistant to osmotic stress. The red blood cell storage lesion is accelerated in previously cryopreserved pRBC after thawing. Biochemical deterioration of thawed and deglycerolized red cells suggests that storage time before transfusion should be limited to achieve similar risk profiles as never-frozen standard liquid storage pRBC units.

Age related changes in inflammatory and regulatory subsets within a healthy population are biased by CMV infection and oldest age.

Abstract Age-related inflammation is detrimental to the host and plays a key initiator of disease. However, it is unclear how patterns of immune cells and particularly inflammatory and regulatory subsets occur within a healthy population. We investigated peripheral blood markers for immunologic subsets and inflammatory markers using samples collected from a local Blood Bank. 319 subjects between 25–81 years were included that had passed standard collection and infection status criteria. Batches of freshly collected whole blood samples were lysed and immediately processed for flow cytometry using 9-color panels of lymphocyte, myeloid, and platelet markers. Data was normalized by fold change to the young (25–35 yr) group included within batches (7 total) compared to older (36–45, 46–55, 56–64, or 65+ year) age groups. Overall donor age was significantly related to weight, blood pressure, and CMV seropositivity by Spearman’s correlation analyses. We observed few immunologic changes between age groups, except significant differences within the 56–64yr age group. These individuals expressed significantly higher levels of antigen-experienced CD45RO+ CD8 T-cells, regulatory TGF-beta+ CD4 T-cells, and plasma B-cells that correlated with CMV seropositivity. The 56–64 age group also expressed higher levels of non-classical monocytes and platelet monocyte aggregates, both related to inflammation. A lack of changes observed in the highest age group (65+) may be due to selection biases from earlier death and disease. Our data supports that responses to chronic viral infections like CMV may be a major regulator of healthy aging, and also reveals potential biases in recruited populations of older adults.

Surviving a Double Whammy.

Surviving a Double Whammy.

Physicochemical Characteristics of Transferon™ Batches.

Transferon, a biotherapeutic agent that has been used for the past 2 decades for diseases with an inflammatory component, has been approved by regulatory authorities in Mexico (COFEPRIS) for the treatment of patients with herpes infection. The active pharmaceutical ingredient (API) of Transferon is based on polydispersion of peptides that have been extracted from lysed human leukocytes by a dialysis process and a subsequent ultrafiltration step to select molecules below 10 kDa. To physicochemically characterize the drug product, we developed chromatographic methods and an SDS-PAGE approach to analyze the composition and the overall variability of Transferon. Reversed-phase chromatographic profiles of peptide populations demonstrated batch-to-batch consistency from 10 representative batches that harbored 4 primary peaks with a relative standard deviation (RSD) of less than 7%. Aminogram profiles exhibited 17 proteinogenic amino acids and showed that glycine was the most abundant amino acid, with a relative content of approximately 18%. Further, based on their electrophoretic migration, the peptide populations exhibited a molecular mass of about 10 kDa. Finally, we determined the Transferon fingerprint using a mass spectrometry tool. Because each batch was produced from independent pooled buffy coat samples from healthy donors, supplied by a local blood bank, our results support the consistency of the production of Transferon and reveal its peptide identity with regard to its physicochemical attributes.

'Red blood transfusion in patients undergoing cardiac surgery'.

The use of allogeneic red blood cells (RBC) is commonplace in cardiac surgery, with reported transfusion rates ranging from 5 to 90 %. [1–4] Despite advantages, RBC transfusion is associated with well-described adverse outcomes. [1–4] Transfusion of RBCs is not only associated with an increased perioperative mortality and morbidity, however; it also results in a longer ICU stay, total hospital stay and increased costs. [2] Even the long-term results are influenced by perioperative RBC transfusions. Koch et al. found a significantly reduced 6-month and late survival in patients undergoing an isolated CABG with transfusion of RBCs. [1] Not only the question of RBC transfusion, but also the number of units of transfused RBCs and the method and duration of storage of RBCs is important. [3, 4] It is no surprise that several studies focus on blood conservation methods and a more judicious use of RBCs, with as prerequisite the identification of preoperative variables associated with an increased need for transfusion. [3, 4] Female gender, impaired left ventricular function, low preoperative haemoglobin, risk and complexity of the operation are the most identified variables with an increased risk for RBC transfusion [3, 4] However, even with the routine use of current blood conservation methods, a proportion of patients undergoing cardiac surgery continue to receive RBC transfusion and are hence exposed to its associated risk [1–4]. There is, therefore, an ongoing need to study further blood conservation methods. In their recent study, [5] Haanschoten et al. focus on reducing the immediate availability of RBCs in cardiac surgery. Since March 2010, the No Elective Red Cells (NERC) program is being used. The only change for their current study is that RBCs are now not directly available in the operating theatre and, when needed, the local blood bank can deliver the cross-matched RBCs within 20 min. So, the aim of this new strategy is to reduce the number of units of RBCs returned to the laboratory and certainly the numbers of those units that can no longer be used because of a reduced quality of the RBCs. This is of course an interesting point in the whole discussion of blood conservation, mostly evaluated only on the percentage of patients without RBC transfusion and /or the number of units transfused. The authors show that in a selected group of patients it is safe to perform cardiac surgery with the immediate availability of RBCs in the operating theatre. [5] Eighty-one percent of the patients did not receive any RBCs and the predictors for the need of blood transfusion are confirming the results of previous studies. [3, 4] However, some interesting points are not discussed in this study. Since the start of the NERC program, what was the percentage of RBCs returned to the blood bank and what was the percentage of units that could no longer be used because of reduced quality? It is emphasised that in all patients who need a blood transfusion, blood was available within 20 min after ordering it. However, was this really so? What was the time between ordering and delivering the blood in the operating theatre? What happens in cases of catastrophic bleeding problems, always possible even in the most simple cardiac procedure? These questions are essential before general conclusions can be made about safety and the effectiveness of this new strategy, no immediate availability of red blood cells, in the NERC program.

Gallbladder cancer incidence in Gwalior district of India: Five-year trend based on the registry of a regional cancer center.

We have reported here the 5-year incidence (2004-2008) of gallbladder cancer (GBC) in North Central India along with its descriptive epidemiology. This provides potential clues for better prevention. The present study has also evaluated the association of ABO blood groups with GBC. The study comprised 742 GBC cases referred to the regional cancer hospital, Gwalior, during 2004-2008. The demographic statistics of Gwalior district was considered to calculate the relative risk and incidence rates. ABO blood group distribution amongst 90,000 healthy subjects registered in the local blood bank during 2002-2007 was taken as controls to study the association of blood groups with GBC. The age-standardized total incidence rate of GBC was calculated to be 7.16/1,00,000. The relative risk of females getting GBC was 2.693 at 95% confidence interval of 2.304-3.151 (P < 0.0001). The females formed 69.5% of total cancer cases, with age-standardized incidence rate of 10/1,00,000. The mean age of male and female GBC cases was found to be 55.4 years (SD = 13, SE = 0.77) and 51.5 years (SD = 12.3, SE = 0.50), respectively. The blood groups A (P = 0.0022) and AB (P < 0.0001) had a positive association with GBC with significant level of differences in comparison to controls. Our study provided an estimate of a 5-year incidence of GBC in North Central India for the first time. With regard to the association of risk factors like obesity, age, and urban living with GBC, the findings of the present study are contradictory to the general opinion. Blood groups A and AB were found to be associated with GBC, which would be provisional for further investigations.

Red blood transfusion in patients undergoing cardiac surgery.

The use of allogeneic red blood cells (RBC) is commonplace in cardiac surgery, with reported transfusion rates ranging from 5 to 90 %. [1–4] Despite advantages, RBC transfusion is associated with well-described adverse outcomes. [1–4] Transfusion of RBCs is not only associated with an increased perioperative mortality and morbidity, however; it also results in a longer ICU stay, total hospital stay and increased costs. [2] Even the long-term results are influenced by perioperative RBC transfusions. Koch et al. found a significantly reduced 6-month and late survival in patients undergoing an isolated CABG with transfusion of RBCs. [1] Not only the question of RBC transfusion, but also the number of units of transfused RBCs and the method and duration of storage of RBCs is important. [3, 4] It is no surprise that several studies focus on blood conservation methods and a more judicious use of RBCs, with as prerequisite the identification of preoperative variables associated with an increased need for transfusion. [3, 4] Female gender, impaired left ventricular function, low preoperative haemoglobin, risk and complexity of the operation are the most identified variables with an increased risk for RBC transfusion [3, 4] However, even with the routine use of current blood conservation methods, a proportion of patients undergoing cardiac surgery continue to receive RBC transfusion and are hence exposed to its associated risk [1–4]. There is, therefore, an ongoing need to study further blood conservation methods. In their recent study, [5] Haanschoten et al. focus on reducing the immediate availability of RBCs in cardiac surgery. Since March 2010, the No Elective Red Cells (NERC) program is being used. The only change for their current study is that RBCs are now not directly available in the operating theatre and, when needed, the local blood bank can deliver the cross-matched RBCs within 20 min. So, the aim of this new strategy is to reduce the number of units of RBCs returned to the laboratory and certainly the numbers of those units that can no longer be used because of a reduced quality of the RBCs. This is of course an interesting point in the whole discussion of blood conservation, mostly evaluated only on the percentage of patients without RBC transfusion and /or the number of units transfused. The authors show that in a selected group of patients it is safe to perform cardiac surgery with the immediate availability of RBCs in the operating theatre. [5] Eighty-one percent of the patients did not receive any RBCs and the predictors for the need of blood transfusion are confirming the results of previous studies. [3, 4] However, some interesting points are not discussed in this study. Since the start of the NERC program, what was the percentage of RBCs returned to the blood bank and what was the percentage of units that could no longer be used because of reduced quality? It is emphasised that in all patients who need a blood transfusion, blood was available within 20 min after ordering it. However, was this really so? What was the time between ordering and delivering the blood in the operating theatre? What happens in cases of catastrophic bleeding problems, always possible even in the most simple cardiac procedure? These questions are essential before general conclusions can be made about safety and the effectiveness of this new strategy, no immediate availability of red blood cells, in the NERC program.

Substitutes of fetal bovine sera for the culture of human corneal limbal stem cells

AbstractPurpose We analyze the potential use of PRGF‐Endoret® and human sera (HS) as a substitute of xenogeneic sera in corneal cell cultures.Methods Plasma rich in growth factors (PRGF) eye drop and HS was obtained at our local blood bank from human blood of healthy donors after informed consent. Briefly PRGF was purified using the Endoret kit in Ophthalmology. HS was obtained according to standardized protocols. Limbal stem cells were obtained from explants of 2‐3mm in diameter of the limbal region of corneoscleral tissue and cultured in culture medium containing 10% PRGF‐Endoret®, HS or fetal bovine serum (FBS) for 7 days. Afterwards, cells were fixed and growth area was quantified. Immunocytochemistry for p.63, ABCG‐2 and cytokeratin was also performed in order to check their immunological markers.Results PRGF‐Endoret®, HS and FBS, cell cultures showed a positive expression in p.63, ABCG‐2 and cytokeratin. However some differences were found between cultures, limbal stem cell cultures supplemented with PRGF‐Endoret® and HS showed a better growth area in contrast with the ones supplemented with FBS. Moreover, there was cellular growth in more of the explants cultured in PRGF‐Endoret® and HS compared to FBS supplemented cultures.Conclusion PRGF‐Endoret® and HS seems to improve the cellular growth and, while maintaining the p63 and ABCG‐2 expression compared to FBS supplemented cultures. Therefore, PRGF‐Endoret® or HS could be used as a substitute of xenogeneic sera, not only as an allogeneic product but also as an autologous product for the culture and expansion of corneal limbal stem cells.

Blood transfusion products contain mitochondrial DNA damage-associated molecular patterns: a potential effector of transfusion-related acute lung injury

BackgroundTransfusion-related acute lung injury (TRALI) is the most frequent and severe complication in patients receiving multiple blood transfusions. Current pathogenic concepts hold that proinflammatory mediators present in transfused blood products are responsible for the initiation of TRALI, but the identity of the critical effector molecules is yet to be determined. We hypothesize that mtDNA damage-associated molecular patterns (DAMPs) are present in blood transfusion products, which may be important in the initiation of TRALI. MethodsDNA was extracted from consecutive samples of packed red blood cells, fresh frozen plasma (FFP), and platelets procured from the local blood bank. Quantitative real-time polymerase chain reaction was used to quantify ≈200 bp sequences from the COX1, ND1, ND6, and D-loop regions of the mitochondrial genome. ResultsA range of mtDNA DAMPs were detected in all blood components measured, with FFP displaying the largest variation. ConclusionsWe conclude that mtDNA DAMPs are present in packed red blood cells, FFP, and platelets. These observations provide proof of the concept that mtDNA DAMPs may be mediators of TRALI. Further studies are needed to test this hypothesis and to determine the origin of mtDNA DAMPs in transfused blood.