Clinical effectiveness of platelets in additive solution treated with two commercial pathogen-reduction technologies.

BACKGROUND: Two noninferiority, randomized, controlled trials were conducted in parallel comparing the safety and efficacy of platelets treated with Intercept or Mirasol pathogen-reduction technologies versus standard platelets. STUDY DESIGN AND METHODS: The primary endpoint was the percentage of hematology patients who developed World Health Organization Grade 2 or greater bleeding. A noninferiority margin of 11% was chosen based on expected Grade 2 or greater bleeding in 20% of controls. The study was closed for financial restrictions before reaching the planned sample size of 828 patients, and an intention-to-treat analysis was conducted on 424 evaluable patients. RESULTS: In the Intercept trial (113 treated vs. 115 control patients), the absolute risk difference in Grade 2 or greater bleeding was 6.1%, with an upper one-sided 97.5% confidence limit of 19.2%. The absolute risk difference in the Mirasol trial (99 treated vs. 97 control patients) was 4.1%, and the upper one-sided 97.5% confidence limit was 18.4%. Neither absolute risk difference was statistically significant. In both trials, posttransfusion platelet count increments were significantly lower in treated versus control patients. Mean blood component use in treated patients versus controls was 54% higher (95% confidence interval, 36%-74%; Intercept) and 34% higher (95% confidence interval, 16%-54%; Mirasol) for platelets and 23% higher (95% confidence interval, 8%-39%; Intercept) and 32% higher (95% confidence interval, 10%-57%; Mirasol) for red blood cells. Unexpected reactions and adverse events were not reported. Mortality did not differ significantly between treated and control patients. CONCLUSION: Although conclusions on noninferiority could not be drawn due to low statistical power, the study provides additional information on the safety and efficacy of pathogen-reduced platelets treated with two commercial pathogen-reduction technologies.

Development and Validation of an Enzymatic Method for Total Cholesterol Analysis Using Whole Blood Spot.

BACKGROUND: High serum cholesterol represents a risk factor for cardiovascular disease. This study aims to quantify total cholesterol in dried blood spot (DBS) by direct enzymatic method. METHODS: Three hundred seventeen blood samples with serum cholesterol level ranging from 81 to 337 mg/dl were collected. DBS were manually prepared, cholesterol was extracted using methanol and analyzed by a manual enzymatic method. DBS cholesterol method was validated for imprecision and extraction efficacy. DBS cholesterol values were correlated (training test) with serum values measured by automated enzymatic method (reference method). The obtained correlation was used for predicting serum cholesterol from DBS analysis of a new sample group (validation test, n = 58). RESULTS: Within-day and between-day coefficient of variation (CV%) were lower than 7.69 and 6.32, respectively. Residual cholesterol in DBS after extraction was 16%. DBS cholesterol and serum cholesterol showed a linear correlation (slope = 0.5217; r = 0.9139) and a bias of -28%. Furthermore, DBS cholesterol values of validation test (n = 58), converted using the training test correlation, were not statistically different compared to the corresponding plasma values (P = 0.9487), and the comparison by Passing and Bablok showed a linear regression with a slope of 1.068 (r = 0.611) and a bias of -0.22%. CONCLUSIONS: The results show that this enzymatic method is suitable to analyze cholesterol in DBS and it could be automated and used for population screening of total blood cholesterol.

Low vitamin D levels are associated with the presence of serum cryoglobulins in patients with chronic HCV infection.

BACKGROUND/AIM: Mixed Cryoglobulinemia (MC) represents the most frequent extrahepatic manifestation of chronic Hepatitis C Virus (HCV) infection. Its pathogenic mechanisms involve HCV-induced chronic stimulation of B-lymphocytes. We aimed to investigate the relationship between serum levels of vitamin D (a regulator of immune response) and the presence of serum cryoglobulins in the setting of HCV infection. PATIENTS AND METHODS: We evaluated the serum concentration of 25(OH)vitamin D and cryoglobulins in 106 patients with chronic HCV infection. RESULTS: Thirty patients (28.3%) showed the presence of serum cryoglobulins. For the cohort overall, the median serum 25(OH)vitamin D level was 10.95 ng/ml. Patients with serum cryoglobulins had significantly lower levels of 25(OH)vitamin D (5.61 ng/ml) than those without (13.65 ng/ml, p=0.029). At multivariate analysis, severe hypovitaminosis [i.e. 25(OH)vitamin D <13 ng/ml] was the only independent predictor of cryoglobulinemia (odds ratio=3.108). CONCLUSION: Severe deficiency of vitamin D was independently associated with mixed cryoglobulinemia in patients with HCV infection.

Prolonged Activated Partial Thromboplastin Time: Difficulties in Discriminating Coexistent Factor VIII Inhibitor and Lupus Anticoagulant.

To review the diagnostic difficulties of a prolonged activated partial thromboplastin time (aPTT) when 2 inhibitors with opposite clinical presentations coexist, we searched MEDLINE from January 1970 to November 2013 using acquired, factor VIII (FVIII), factor IX, hemophilia A and B, inhibitor, lupus anticoagulant (LA), antiphospholipid, anticardiolipin, anti-ß2-glycoprotein I, antibodies, syndrome, bleeding, and thrombosis. We identified 13 articles for a total of 15 cases of possible coexistence of FVIII inhibitor and LA. The presenting clinical manifestation was thrombosis in 6 cases and bleeding in 9 cases. Activated partial thromboplastin time was the presenting laboratory abnormality in all cases, and first-line investigations suggested the coexistence of LA and acquired FVIII inhibitor. None of the articles addressed the diagnostic accuracy of the screening tests by performing "second line" assays. We reviewed the diagnostic pitfalls of the cases under study and provide some guidance for alternative tests when facing a prolonged aPTT that may have a double meaning.

Human platelet antigen-3 genotype predicts platelet count in patients with HCV infection.

BACKGROUND/AIM: A low platelet count is one of the most sensitive tests for cirrhosis detection in patients with hepatitis C virus (HCV) infection. We evaluated whether the human platelet antigen (HPA) genotype could predict platelet count in HCV-positive patients. MATERIALS AND METHODS: We genotyped the HPA 1, 2, 3, 5 and 15 polymorphisms in consecutive patients with HCV infection. RESULTS: Out of the 56 patients enrolled, 56.1% had liver cirrhosis. The mean platelet count was significantly lower in those with HPA1aa genotype than in those with HPA1ab/bb genotype. Platelet count did not differ among the other HPA polymorphisms. However, at logistic regression analysis, only the HPA3aa genotype and liver cirrhosis were independent predictors of a low platelet count. CONCLUSION: HPA3aa is an independent factor for a low platelet count in this cohort of patients with HCV chronic infection regardless of disease stage.

Fetoneonatal alloimmune thrombocytopenia (FNAIT): our experience.

OBJECTIVE: Fetoneonatal alloimmune thrombocytopenia (FNAIT) is a relatively rare clinical syndrome characterized by marked thrombocytopenia shortly after birth. It occurs when fetal platelets are destroyed, after sensitization, by a transplacental passage of maternal antibodies directed against a fetal platelet alloantigen inherited from the father. This article reviews some pathophysiologic and clinical aspects of FNAIT. METHODS: We also present our experience with the management of 12 newborns affected with a symptomatic form of this disorder in order to verify what would be the best diagnostic and therapeutic protocols. RESULTS: Antibody identification in maternal serum showed 9 anti-HPA-1a (75% of cases), 2 anti-HPA-1b (17%) and 1 anti-HPA-1a+anti-Gp IV+anti-HLA class I (8%). CONCLUSION: Sixteen human platelet alloantigen (HPA) systems have been identified, six major (from HPA 1 to 5 and HPA 15) and ten rare or private, each composed of two allelic antigens (named "a" or "b", according to major or minor frequency in the population). All HPA systems, including private or low frequency, may play a role in determining FNAIT. Unfortunately FNAIT cannot be prevented, in fact no one of maternal parameters is predictive of thrombocytopenia or its magnitude.

The metastasis-associated 67-kDa laminin receptor is involved in G-CSF-induced hematopoietic stem cell mobilization.

The 67-kDa laminin receptor (67LR) is a nonintegrin cell-surface receptor with high affinity for laminin, which plays a key role in tumor invasion and metastasis. We investigated the role of 67LR in granulocyte colony-stimulating factor (G-CSF)-induced mobilization of CD34+ hematopoietic stem cells (HSCs) from 35 healthy donors. G-CSF-mobilized HSCs, including CD34+/CD38- cells, showed increased 67LR expression as compared with unstimulated marrow HSCs; noteworthy, also, is the fact that the level of 67LR expression in G-CSF-mobilized HSCs correlated significantly with mobilization efficiency. During G-CSF-induced HSC mobilization, the expression of laminin receptors switched from alpha6 integrins, which mediated laminin-dependent adhesion of steady-state human marrow HSCs, to 67LR, responsible for G-CSF-mobilized HSC adhesion and migration toward laminin. In vitro G-CSF treatment, alone or combined with exposure to marrow-derived endothelial cells, induced 67LR up-regulation in marrow HSCs; moreover, anti-67LR antibodies significantly inhibited transendothelial migration of G-CSF-stimulated marrow HSCs. Finally, G-CSF-induced mobilization in mice was associated with 67LR up-regulation both in circulating and marrow CD34+ cells, and anti-67LR antibodies significantly reduced HSC mobilization, providing the first in vivo evidence for 67LR involvement in stem-cell egress from bone marrow after G-CSF administration. In conclusion, 67LR up-regulation in G-CSF-mobilized HSCs correlates with their successful mobilization and reflects its increase in marrow HSCs, which contributes to the egress from bone marrow by mediating laminin-dependent cell adhesion and transendothelial migration.

Spleen enlargement following recombinant human granulocyte colony-stimulating factor administration for peripheral blood stem cell mobilization.

BACKGROUND AND OBJECTIVES: Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is widely used to mobilize peripheral blood stem cells (PBSC) for autologous or allogeneic transplants. Such treatment may cause spleen enlargement; exceptionally, spontaneous spleen rupture has been reported. We investigated changes in spleen size during stem cell mobilization. DESIGN AND METHODS: We evaluated spleen size, comparing palpation with ultrasound (US)-evaluated longitudinal diameter and volume, in 13 healthy donors and 22 patients with a hematological malignancy who were undergoing PBSC mobilization with rhG-CSF-including regimens. RESULTS: Intraobserver and interobserver variability of US-calculated spleen volume was very low; the correlation between the volume calculated by US and that measured by 3-dimensional computed tomography was excellent. During mobilization, spleen enlargement was detected by palpation in 17% of subjects, by US-measured longitudinal diameter in 60%, and by US-calculated volume in 91%. The median increase in spleen volume was 300 mL (range, 54-820; p<0.001) in healthy donors and 135 mL (range, 0-413; p=0.004) in the group of patients; the enlargement correlated with white blood cell count elevation (p=0.016) but not with circulating CD34+ cells. One month after the last administration of rhG-CSF, the median decrease was 160 mL (range, 35-800) in healthy donors and 58 mL (range, 0-310) in patients. INTERPRETATION AND CONCLUSIONS: When evaluated by sensitive methods, rhG-CSF caused spleen enlargement in almost all individuals treated. US-calculated volume proved to be an excellent method, much better than longitudinal diameter, for detecting non-palpable splenomegaly induced by rhG-CSF.