Rapid exclusion of the diagnosis of immune HIT by AcuStar HIT and heparin-induced multiple electrode aggregometry.

BACKGROUND: Accurate diagnosis of heparin-induced thrombocytopenia (HIT) is essential but remains challenging. We have previously demonstrated, in a retrospective study, the usefulness of the combination of the 4Ts score, AcuStar HIT and heparin-induced multiple electrode aggregometry (HIMEA) with optimized thresholds. OBJECTIVES: We aimed at exploring prospectively the performances of our optimized diagnostic algorithm on suspected HIT patients. The secondary objective is to evaluate performances of AcuStar HIT-Ab (PF4-H) in comparison with the clinical outcome. METHODS: 116 inpatients with clinically suspected immune HIT were included. Our optimized diagnostic algorithm was applied to each patient. Sensitivity, specificity, negative predictive value (NPV), positive predictive value (PPV) of the overall diagnostic strategy as well as AcuStar HIT-Ab (at manufacturer's thresholds and at our thresholds) were calculated using clinical diagnosis as the reference. RESULTS: Among 116 patients, 2 patients had clinically-diagnosed HIT. These 2 patients were positive on AcuStar HIT-Ab, AcuStar HIT-IgG and HIMEA. Using our optimized algorithm, all patients were correctly diagnosed. AcuStar HIT-Ab at our cut-off (>9.41 U/mL) and at manufacturer's cut-off (>1.00 U/mL) showed both a sensitivity of 100.0% and a specificity of 99.1% and 90.4%, respectively. CONCLUSION: The combination of the 4Ts score, the HemosIL® AcuStar HIT and HIMEA with optimized thresholds may be useful for the rapid and accurate exclusion of the diagnosis of immune HIT.

Platelet microparticle generation assay: a valuable test for immune heparin-induced thrombocytopenia diagnosis.

BACKGROUND: Early diagnosis of immune heparin-induced thrombocytopenia (HIT) is essential to improve clinical outcome but remains challenging. The release of platelet microparticles (PMPs) is considered of major pathophysiological significance. OBJECTIVES: The aim of this study was to evaluate performances of PMP generation assay (PMPGA) compared to clinical outcome to diagnose HIT. The second objective was to compare PMPGA with performances of (14)C-serotonin release assay (SRA) on the same series of patients. METHODS: Sera of 53 HIT-suspected patients were retrospectively incubated with citrated-whole blood from healthy donors with 1IU and 500IU/ml of unfractionated heparin (UH). PMPGA was performed using FACSAria® flow cytometer. The clinical diagnosis was established by two blinded independent investigators analysing in a standardized manner the patient's medical records. Performances of PMPGA and SRA (n=53) were evaluated using ROC curve analysis with clinical outcome as reference. RESULTS: In positive HIT patients, PMPs expressing phosphatidylserine are generated with low UH concentration whereas PMP rate decreases significantly in presence of high UH concentration. Using clinical outcome as reference, sensitivity and specificity of PMPGA reached 88.9% (95% CI: 50.7-99.4) and 100.0% (95% CI: 90.0-100.0). Sensitivity and specificity of (14)C-SRA were 88.9% (95% CI: 50.7-99.4) and 95.5% (95% CI: 83.3-99.2). CONCLUSIONS: PMPGA is a rapid and reliable assay for HIT diagnosis. PMPGA showed good correlation with (14)C-SRA performances and predominately with clinical outcome.

Assessment of the performances of AcuStar HIT and the combination with heparin-induced multiple electrode aggregometry: a retrospective study.

BACKGROUND: Early diagnosis of immune heparin-induced thrombocytopenia (HIT) is challenging. HemosIL® AcuStar HIT and heparin-induced multiple electrode aggregometry (HIMEA) were recently proposed as rapid diagnostic methods. OBJECTIVES: We conducted a study to assess performances of AcuStar HIT-IgG (PF4-H) and AcuStar HIT-Ab (PF4-H). The secondary objective was to compare the performances of the combination of Acustar HIT and HIMEA with standardised clinical diagnosis. METHODS: Sera of 104 suspected HIT patients were retrospectively tested with AcuStar HIT. HIMEA was performed on available sera (n=81). The clinical diagnosis was established by analysing in a standardized manner the patient's medical records. These tests were also compared with PF4-Enhanced®, LTA, and SRA in subsets of patients. Thresholds were determined using ROC curve analysis with clinical outcome as reference. RESULTS: Using the recommended thresholds (1.00AU), the negative predictive value (NPV) of HIT-IgG and HIT-Ab were 100.0% (95% CI: 95.9%-100.0% and 95.7%-100.0%). The positive predictive value (PPV) were 64.3% (95% CI: 35.1%-87.2.2%) and 45.0% (95% CI: 23.2%-68.6%), respectively. Using our thresholds (HIT-IgG: 2.89AU, HIT-Ab: 9.41AU), NPV of HIT-IgG and HIT-Ab were 100.0% (95% CI: 96.0%-100.0% and 96.1%-100.0%). PPV were 75.0% (95% CI: 42.7%-94.5%) and 81.8% (95% CI: 48.3%-97.7%), respectively. Of the 79 patients with a medium-high pretest probability score, 67 were negative using HIT-IgG (PF4-H) test at our thresholds. HIMEA was performed on HIT-IgG positive patients. Using this combination, only one patient on 79 was incorrectly diagnosed. CONCLUSION: Acustar HIT showed good performances to exclude the diagnosis of HIT. Combination with HIMEA improves PPV.

Different periods of feed restriction before compensatory growth in Belgian Blue bulls: II. Plasma metabolites and hormones.

Plasma metabolites and hormones were studied in 16 double-muscled Belgian Blue bulls maintained at low growth (.5 kg/d) for 114 (G2), 243 (G3), or 419 (G4) d (low growth period, LGP) before fattening (rapid growth period, RGP). Animals from the control group (CG) were fed a diet high in energy and protein. The animals from G2, G3, and G4 were fed a restricted amount of a diet low in energy and protein during LGP and the same diet as CG during RGP. Plasma glucose, alpha-amino nitrogen (AAN), NEFA, urea, creatinine, thyroxine (T4), 3,3',5'-triiodothyronine (T3), and IGF-I were measured in blood samples taken fortnightly. Plasma GH and insulin (I) profiles were measured in serial blood samples obtained at three times during growth. The RGP was characterized by an initial compensatory growth, by higher plasma glucose, AAN, and urea levels, and by lower plasma NEFA and creatinine levels. Plasma GH concentration decreased after refeeding. Plasma T4 increased linearly during refeeding, as opposed to T3, which showed a different profile in each group. Plasma IGF-I showed a curvilinear increase during RGP and reached a plateau after 3 mo in each compensating group. In G4, changes of plasma metabolites and hormones differed often distinctly from G2 or G3. During refeeding, higher nutrient supply improved the functionality of the somatotropic axis and increased the concentration of anabolic hormones, allowing rapid muscle deposition. However, animals underfed the longest period behaved differently from the other groups, possibly because they reached a more complete sexual maturity.

Comparison of hindquarter metabolite uptakes in Belgian Blue double-muscled bulls at maintenance or during fattening.

Metabolism of muscle growth in the hindquarter was investigated by the arterio-venous difference (AVD) technique in Belgian Blue double-muscled type bulls at maintenance or at fattening. The bulls were fitted with an aortic ultrasonic blood flow probe and with catheters in the aorta and vena cava. They were offered a diet allowing for maintenance (MP) during a period of 15 d, at the end of which measurements were made over 3 d. Bulls were then given a fattening diet (FP) and the measurements were repeated. Arterial blood flow was approximately 1 L/min greater when the bulls were standing than when lying. Blood flow was 2 L/min higher during FP than during MP. The AVD and uptake of glucose were maximal at 1400 and 1600. Uptake of alpha-amino nitrogen decreased immediately after a meal. The increase in glucose from MP to FP fitted very well with the calculated energy needs for muscle growth. The AVD and uptake of alpha-amino nitrogen, total amino acids, and total nonessential amino acids were negative during MP and positive and significantly higher during FP. There was also a significant increase in AVD and uptake of essential and branched-chain amino acids when the bulls were changed from MP to FP. When changing from maintenance to fattening, the incremental glucose and amino acid hindquarter uptake provided energy and supply for muscle protein accretion, respectively. The level of alanine transamination was also sharply reduced.