Hereditary thrombocytopenia with platelet sialic acid deficiency and mutations in the GNE genes.

BACKGROUND: The inherited macrothrombocytopenias are rare disorders and the underlying cause can be identified in many cases but in some, this can remain enigmatic. Platelet transfusions are often administered during hemorrhagic events. METHODS: A patient with previously unexplained inherited macrothrombocytopenia with a platelet count between 3-20 × 109 /L is described in which studies were performed using exome sequencing (ES) and platelet flow cytometry. RESULTS: Both the hemoglobin and white cell counts were normal. ES revealed two suspicious variants, one likely pathogenic and one a variant of uncertain significance, in the UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) gene, and flow cytometry showed diminished expression of surface platelet sialic acid (about 5%) but normal red cell sialic acid. The Thrombopoietin (TPO) level was low, and the patient responded to TPO-mimetic treatment with an increase in the platelet count. CONCLUSION: Two variants in the GNE gene were able to be upgraded to pathogenic with apparently restricted expression to the megakaryocyte lineage. Platelet transfusion may be avoided in these patients with TPO-mimetic treatment.

Discordance in tests used to detect inhibition of the P2Y12 receptor in patients undergoing interventional neuroradiology procedures.

BACKGROUND: Clopidogrel is an inhibitor of the P2Y12 platelet receptor but testing to demonstrate a drug effect is controversial since there are often discordant results between different tests methods. METHODS: Samples from patients taking clopidogrel prior to intracranial flow-diversion procedures were tested using light transmission aggregometry (LTA), whole blood impedance aggregometry (WBIA) and the VerifyNow device (VND). Samples were classified as concordant if all test results were either responsive (inhibition) or resistant. Discordant results were separated using the VND into those with a responsive versus a resistant test result. RESULTS: Samples from 96 patients were studied. Concordance for all three tests was seen in 53/96 (55%) of samples, of which 41 (43%) were responsive and 12 (12%) were resistant. Discordance was observed in 43 samples (45%), 37 (28%) of which were caused by responsive VND and either a resistant WBIA or LTA and 6 (7%) of which were caused by a resistant VND but a responsive test result using either WBIA or LTA. These two discordant groups differed in both platelet count and hematocrit, but no such difference was present between the two concordant groups. CONCLUSION: Discordance in P2Y12 inhibition testing may be partly explained by sample platelet count and hematocrit.

In vitro comparison of CPD whole blood with conventional blood components.

INTRODUCTION: Resuscitation of severely injured trauma patients is commonly performed using red blood cells in additive solution supplemented with plasma and platelet concentrates. There is an increasing interest in the use of low anti-A titer Group O whole blood (LTOWB) in the early management of the resuscitation. It is unclear whether clinical outcome is improved using this approach. METHODS: Expired units of CPD-LTOWB were studied on Day 22 and expired units of thawed plasma on Day 6 and Day 7. LTOWB was assessed for hemoglobin content, clotting factor levels and platelet numbers and function using thromboelastography (TEG) and impedance aggregation. Assays of fibrinogen and FV, FVIII, FVII and FX were performed on the expired plasma. The LTOWB hemoglobin was compared to red cells in additive solution (AS-RBCs) and the clotting factor levels to those of expired thawed plasma. Platelet function was compared to fresh whole blood samples from healthy subjects. RESULTS: LTOWB contained slightly more hemoglobin than the AS-RBCs (Medians, 66 v 59 G), and the plasma content of fibrinogen was similar. Other clotting factors were reduced by approximately 15% except for FVIII which was 30% less. Both TEG and impedance aggregometry showed evidence of residual platelet function despite the prolonged period of refrigerator storage. CONCLUSION: LTOWB contains higher hemoglobin and adequate clotting factors, and residual platelet function is demonstrated indicating that this product would be expected to be at least equivalent to a single unit of each of the conventional components commonly used in trauma resuscitation.

Laboratory Methods in the Assessment of Hereditary Hemostatic Disorders.

In patients presenting with a suspect hereditary bleeding disorder a detailed bleeding history is first obtained. Testing proceeds in a tiered manner with platelet count, platelet morphology, platelet histogram, PFA-100, fibrinogen, prothrombin time, and activated partial thromboplastin time. More detailed testing includes von Willebrand factor, individual clotting factor assays, and platelet function testing. Next, testing for a dysfibrinogenemia, FXIII, or a fibrinolytic defect is considered. Hemostatic abnormality is not demonstrated in a fraction of patients. An approach to management in these patients, such as desmopressin or antifibrinolytic therapy, may be required and empiric use of blood component therapy is discouraged.

Red Cell Exchange as Adjunctive Therapy for Babesiosis: Is it Really Effective?

Human babesiosis is a parasitic disease prevalent in the Northeastern and Midwestern United States (US). Treatment with antibiotics is the standard of care but red cell exchange (RCE) has been used as an adjunctive treatment in more severe disease. Data for the efficacy of RCE in the treatment of babesiosis has been based on case reports and case series. An English language literature search was conducted for cases of babesiosis treated with RCE since 1980 and relevant laboratory and clinical outcome data were extracted. Similar data were obtained on severe cases of babesiosis referred for RCE in our hospitals in the time period 2000 to 2020. Fifty reports including forty-one individual case reports and nine case series were retrieved. There were 108 patients that underwent RCE with an overall mortality rate of 20%. Some patients had more than one RCE. The patients varied in the level of anemia and evidence of compromise of renal or pulmonary function. The pre-RCE level of parasitemia varied between 1.7% to 85% with the vast majority >10%. The post-RCE level of parasitemia varied between 1% to 10%. Since 2000, 32 patients were referred for RCE in our hospitals and RCE was performed on 23 of 32. There were more patients treated with RCE in the second decade as compared to the first decade, 19 versus 4 respectively. The overall mortality was 22% similar to the national data. Comparing the cohort treated with RCE to the 9 patients who were treated only with antibiotics, there were similar levels of parasitemia and laboratory parameters. The overall number of days needed to achieve a parasite count <1% was similar between the two cohorts and mortality for the antibiotics only cohort was 0%. More than 40 years after the first reported case of RCE in severe babesiosis it cannot be concluded that this adjunctive therapy favorably influences the clinical outcome. Since there is largely equipoise, a registry of severe patients treated with or without RCE could identify a benefit or otherwise.

Sulfamethoxazole-induced thrombocytopenia masquerading as posttransfusion purpura: a case report.

BACKGROUND: Drug-induced immune thrombocytopenia (DITP) is a rare clinical disorder characterized by accelerated platelet (PLT) clearance in the presence of drug-dependent antibodies. Distinguishing DITP from other immune-mediated disorders such as posttransfusion purpura (PTP) and autoimmune thrombocytopenia can represent a clinical challenge. CASE REPORT: A 68-year-old male with no prior transfusion history presented to the emergency department (ED) with dyspnea, epistaxis, and severe thrombocytopenia (<10 × 10(9)/L) 12 days after discharge from a hospital admission for a coronary artery bypass graft. Evaluation of the degree of thrombocytopenia and the temporal association between the peri- and postoperative receipt of multiple transfusions and the acute decrease in PLT count indicated PTP as a possible cause of the severe thrombocytopenia. Treatment with 1 g/kg intravenous immunoglobulin (IVIG) was initiated and followed by a rapid 48-hour increase in the PLT count. PLT antibodies lacking serologic specificity were subsequently identified in a sample collected upon presentation. Two weeks later he again presented to the ED with epistaxis and severe thrombocytopenia (<10 × 10(9)/L). Clinical history now revealed that the patient had been treated with trimethoprim-sulfamethoxazole by his primary care physician after his first hospitalization for a "cellulitic-appearing" leg and again before his final presentation for surgical site erythema and edema. IVIG was administered again with a rapid return of PLT count to baseline. Sulfamethoxazole-dependent PLT antibodies were subsequently identified in the original patient sample. CONCLUSION: This case report documents a case of IVIG-responsive DITP initially misdiagnosed as PTP, highlighting the clinical overlap of these immunologic-mediated phenomena.

The effect of filtration on residual levels of coagulation factors in plasma.

Leukoreduced blood components are commonly manufactured via filtration. There are specifications for the residual leukocyte content of any final cellular blood component but not for residual clotting factors. Leukoreduced and nonleukoreduced platelet-poor plasma products were manufactured from filtered vs unfiltered platelet-rich plasma, respectively, using platelet leukoreduction filters. The leukoreduced plasma showed lower levels of factor VIII (75% ± 16% vs 88% ± 18%, P ≤ .05), factor XI (86% ± 9% vs 96% ± 10%, P ≤ .01) and factor VII (87% ± 14% vs 98% ± 11%, P ≤ .01). No difference was seen with factor X, factor V, or fibrinogen. Plasma filtered through a whole blood filter showed a reduction in factor V (105% ± 12% vs 124% ± 10%, P ≤ .01) but a minimal reduction in factor VIII (80% ± 5% vs 82% ± 6%, P = .04). Filtration can alter the residual levels of clotting factors to a variable extent in manufactured plasma, most noticeably factors V, VII, VIII, and XI.

The effect of specimen hemolysis on coagulation test results.

Hemolyzed specimens are rejected for coagulation testing based on concerns of artifactual interference. Prothrombin time (PT), activated partial thromboplastin time (aPTT), and selected factor assay test results for consecutive pairs of hemolyzed and subsequently recollected (mean, 72 minutes later) nonhemolyzed patient specimens were compared. Specimens from healthy human subjects were subjected to mechanically induced hemolysis, and PT and aPTT results compared with concurrently drawn nonhemolyzed control samples. In 50 paired patient specimens, there were statistically significant differences in PT (15.8 +/- 8.4 vs 16.3 +/- 8.7 seconds, P < .01) and aPTT (31.6 +/- 18 vs 32.5 +/- 19 seconds, P < .01) between hemolyzed and nonhemolyzed specimens, respectively. Specimens from healthy subjects showed no difference in PT and a minor difference in aPTT. A policy of rejecting hemolyzed specimens for coagulation tests should be revisited because the observed difference, when present, is unlikely to be considered clinically meaningful.

Prestorage pooled whole-blood-derived leukoreduced platelets stored for seven days, preserve acceptable quality and do not show evidence of a mixed lymphocyte reaction.

BACKGROUND: Prestorage pooling of whole-blood-derived PCs (WBD-PCs) would be advantageous to transfusion services in that it would make the product available in a more timely manner, reduce wastage of untransfused pools, and simplify bacterial screening by allowing testing of the pool rather than each single PLT concentrate (PC). STUDY DESIGN AND METHODS: Four to six individual leukoreduced PCs were pooled into a 1.5-L CLX-HP PLT storage bag using a sterile connecting device. Controls were individual prestorage leukoreduced PCs that were stored as single products. Products were sampled on Days 5 and 7 for measures of PLT quality; coagulation, fibrinolytic and complement activation; and for evidence of a mixed lymphocyte reaction. RESULTS: The pH level was well maintained to Day 7 with no prestorage pool having a pH below 6.7. Day 7 studies showed no evidence of coagulation or difference in complement activation. F1.2 levels did not differ between Days 5 and 7, but a 10- to 15-percent increase in C3a des-Arg was observed between these days in all product types. Day 7 activated lymphocyte surface markers (CD69, CD71, HLA-DR) were all at lower limits of detection in the prestorage pooled products, and levels of supernatant cytokines were either not different between product types on either study day or, if different, were lower in the prestorage pooled products. CONCLUSION: There is no evidence of a deterioration in quality, activation of coagulation or complement, or a mixed lymphocyte reaction attributable to the prestorage pooling process with up to 7 days of storage.