Combination of high on-treatment platelet aggregation and low deaggregation better predicts long-term cardiovascular events in PCI patients under dual antiplatelet therapy.

High on-treatment platelet reactivity is associated with short-term major cardiovascular (CV) events in patients undergoing percutaneous coronary intervention (PCI). Maximum and final aggregation assessed by light transmission aggregometry (LTA) have both been used to predict short-term outcome after PCI, however their long-term prognostic impact remains controversial. There is currently no information regarding the prognostic role of deaggregation and its added value in combination with established aggregation parameters. About 1279 patients with symptomatic coronary artery disease (CAD) undergoing PCI were enrolled in this monocentric study. On-treatment platelet aggregation under clopidogrel maintenance therapy, as well as deaggregation was determined by maximum and final aggregation (5 min after adding of the agonist). Deaggregation was defined as slope of the tangent between Aggmax +0.5 min. Primary endpoints were the composite of myocardial infarction, stroke, and CV death or stent thrombosis according to the ARC criteria. Low deaggregation, defined as values in the lowest tertile (<1.5), was more frequent in patients with acute coronary syndromes (ACS) compared to patients with stable angina pectoris (SAP), ACS: 29.6% vs. SAP: 22.0%, p = 0.001. The combination of high on-treatment platelet reactivity, defined by the upper tertile of Aggmax and low deaggregation, was associated with significantly increased risk for combined long-term CV events. The combination of low deaggregation and high on-treatment platelet reactivity is associated with higher risk for recurrent events in patients with CAD undergoing PCI. Thus, deaggregation might be a more sensitive parameter providing added value in terms of risk prediction for long-term recurrent CV events in relation with established aggregation parameters.

Hemostasis in Danio rerio: is the zebrafish a useful model for platelet research?

New scientific models have been established in the past few years to identify novel factors of hemostasis and thrombosis and to analyze their function in greater detail. One fairly new animal model is the zebrafish, Danio rerio, which shares most of the central factors of platelet adhesion, activation, aggregation and release reaction with humans. Examples include GPIIb-IIIa, many other integrins, coagulation factors, inflammatory and cytokine-like proteins as well as arachidonic acid metabolism enzymes. Yet the zebrafish genome has undergone a teleost-specific genome duplication, causing the existence of duplicated paralogues in some instances, and a few genes have not been identified in the zebrafish genome. Taken together the high fecundity of the zebrafish, the possibility to observe transparent developing embryos in real time, the availability of a large number of mutants and transgenics as well as the possibility to knock down gene function by microinjection of morpholino antisense oligonucleotides and the similarity of the hemostatic system are important assets of the zebrafish, promising that it will be an attractive model to study thrombocyte function, thrombosis and hemostasis. This review provides an overview of the central factors of thrombocyte function identified so far in the zebrafish genome and a compilation of methods and tools available for the study of thrombocyte development and function in zebrafish.

Defective sperm decondensation: a cause for fertilization failure.

The study aimed to evaluate the role of chromatin packaging (CMA3 staining), sperm morphology during sperm-zona binding, sperm decondensation and the presence of polar bodies in oocytes that failed in vitro fertilization (IVF). The percentage CMA3 staining categorized the data into three groups, < 44%, n = 10; > or = 44-59%, n = 10; and > or = 60%, n = 29. Morphology groups were < or = 4% (n = 11); > 4-14% (n = 19); and > 4% (n = 19). One hundred and seventy-two oocytes that failed IVF were evaluated for sperm-zona binding, ooplasma penetration and sperm decondensation. Odds ratio analyses indicated that being in the > or = 60% CMA3 staining group resulted in a 15.6 fold increase in the risk of decondensation failure, relative to CMA3, staining of < 44%. For morphology, there was a 2.17 fold decrease in the risk of fertilization failure in the morphology group with > 4-14% normal cells, while it increased 2.45 fold for the morphology group with < or = 4% normal cells. Using CMA3 fluorescence to discriminate, 51% of the oocytes in the group with elevated CMA3 fluorescence had no sperm in the ooplasma compared to 32% and 16% penetration failure in the CMA3 staining groups > or = 44-59% and < 44%, respectively. Sperm chromatin packaging quality and sperm morphology assessments are useful clinical indicators of human fertilization failure. Immunofluorescence techniques could be used to provide a clear diagnosis of failed fertilization.

Chromatin packaging as an indicator of human sperm dysfunction.

PURPOSE: Understanding the causes of fertilization failure is an important research field in assisted reproductive programs. The present study aimed to evaluated the possible relationship between chromatin packaging quality (CMA3 staining) and (i) normal morphology and (ii) its ability to predict the functional integrity of spermatozoa in both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment programs. METHODS: Semen of 140 men from IVF and ICSI couples were analyzed for sperm concentration, motility, morphology, and chromatin packaging (CMA3). For CMA3 classification, two cutoff values were used, namely, 44.5% +/- 13 and 1 SD above the mean, i.e., 57.5% (rounded off to 60%). IVF and ICSI data were stratified using three basic cutoff values for CMA3 staining, namely, < 44%, > 44-60%, and > 60%. RESULTS: Based on CMA3 results patients were divided into four groups, namely, group A, < 44% CMA3 (n = 15, IVF); group B, > or = 44% and < 60% CMA3 (n = 39, IVF); group C, > or = 60% CMA3 (n = 45 IVF); and group D, > or = 60% CMA3 (n = 41 ICSI). During receiver operator characteristic analyses the estimated cutoff value for CMA3 staining, to distinguish between < 4% and > or = 4% morphology groups, was 60%. The area under the curve was 0.89, sensitivity of 75%, and specificity of 100%. When IVF rates of > 60% and < 60% were used, the optimal CMA3 value for prediction of fertilization success again was recorded at 60%. The area under the curve was 0.76, sensitivity of 81.5%, and specificity of 63.6%. CONCLUSIONS: Chromatin packaging assessments should be included as a complementary assay to the sequential diagnostic approach of the male-factor patients.

Seminal carnitine, epididymal function and spermatozoal motility.

The carnitine content of semen from azoospermic subjects and subjects with either impaired motility or forward progression is reported. Within the azoospermic group, the values were not significantly different between those with testicular failure or obstruction. The patients with impaired motility (less than 40%) or impaired forward progression (less than 3) had a lower mean seminal carnitine content than those with good motility and forward progression (P less than 0.05). Seminal carnitine concentration was not an index of epididymal patency and could not be regarded as useful for locating the level of obstruction in the epididymis. Since carnitine plays an important role in the maturation process when sperm acquire motility, it would seem that seminal carnitine might be an indicator of epididymal dysfunction.