Overview of pleiotropic effects of platelet P2Y12 receptor inhibitors.

Dual antiplatelet therapy consisting of one of the P2Y12 receptor inhibitors in conjunction with aspirin is the mainstay of treatment for patients with acute coronary syndromes (ACS) and those undergoing percutaneous coronary interventions (PCI). In recent years, multiple extra-platelet features of P2Y12 receptor antagonists have been reported in numerous clinical trials. The aim of this review is to summarise reported pleiotropic effects of clopidogrel, prasugrel, ticagrelor and other P2Y12 receptor blockers. We included observations made both in human and in animal models, together with proposed mechanisms of action for described features. If confirmed in randomised studies and properly applied to everyday practice, the observed extra-platelet actions could enable us to improve efficacy of ACS and post-PCI treatment, as well as to confine mortality and occurrence rate of cardiovascular events.

Technical note: Selecting the best references in gene expression experiments in liver of cows receiving glucogenic supplements during the transition period.

Measuring gene expression is a commonly used method to monitor the reaction of cells and tissues to changing nutritional or physiological conditions. Selection of appropriate reference genes is a crucial point in gene expression experiments using real-time PCR techniques. Expression of the "ideal" reference gene should not be affected by the experimental treatments or physiological state of the tissue, organ, or the whole organism. Many programs are available from which to choose the most stable reference gene. In this study, 4 algorithms--ΔCt, BestKeeper, NormFinder, and geNorm--were used to assess the expression stability of 5 candidate reference genes: ß-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein S9 (RPS9), ribosomal protein L32 (RPL32), and TATA-box-binding protein (TBP), for use in an experiment aimed at measuring gene expression in the liver of cows fed glucogenic supplements in the transition from pregnancy to lactation. The results demonstrated that RPS9 and RPL32 were the most stably expressed in the liver under the conditions of the present experiment; the least stably expressed was ACTB.

Long-term biochemical evaluation of the androgen receptor pathway in males with disorders of sex development.

PURPOSE: Disorders of sex (DSD) development represent a serious condition. Most of the underlying mechanisms remain unclear. Disturbances within the androgen receptor (AR) pathway frequently account for 46 XY-DSDs. The individual gender-related outcome often is unsatisfactory. We present a long-term AR gene-mutation-associated follow-up in a group of 46 XY-DSD patients. METHODS: Twenty patients (46 XY) who underwent genitoplasty in infancy or early childhood were retrospectively identified. Median follow-up after surgery was 16 years. All were undervirilized at initial presentation. Thirteen had female gender assignment, and 7 were raised as males. A genital skin biopsy and subsequent fibroblast cultures were done. The specific binding of dihydrotestosterone, the thermostability of the receptor hormone complex, and 5-α-reductase activity were measured. AR gene mutations were detected by direct sequencing. The individual outcome was correlated with specific AR mutations. RESULTS: AR point mutations were detected in 12, 7 were previously unknown. There was no specific androgen binding in 3, reduced affinity in 9, and normal binding in 8 patients. 5-α-Reductase activity was normal in 15, reduced in 4 and completely absent in 1 patient. CONCLUSIONS: Retrospective evaluation revealed previously unknown and established AR gene mutations being associated with a distinct long-term outcome. Identification of the molecular mechanisms causing DSD will likely improve timely diagnosis and therapy. Exact characterization of AR activation and function may offer a treatment modality in affected patients. These data may allow us to give prognostic estimations on the individual outcome adding objective criteria for gender assignment in 46 XY-DSD patients.

[CRT test used for the evaluation of enamel solubility in permanent teeth caries]. / Zastosowanie testu CRT w ocenie rozpuszczalnosci szkliwa w próchnicy zebów stalych.

The aim of the study was determination of the microsolubility of permanent teeth enamel by the CRT test depending on the degree of caries intensity. Indirectly, it was tried to obtain an answer to the question whether the test could be used for the assessment of caries susceptibility. Forty-six patients of either sex aged 18 to 30 years were studied. The state of caries was determined by the CDF index. Enamel solubility was determined by the CRT test on the surface of permanent teeth--incisors, canine teeth, second premolars and first molars. In all, 460 surface were examined. After the test 75% fluoride paste was rubbed into the surface. The obtained results of the CRT test were collected in two groups depending on the increasing value of the CDF index. The statistical analysis was done using the correlation coefficient and Student's t test. No statistically significant correlation was found between enamel microsolubility expressed by the CRT test and the degree of caries intensity determined by the CDF index. The results obtained by this method could not confirm the view that the CRT test is useful for determination of enamel susceptibility to caries development.

Synthesis of S-lactoyl-glutathione using glyoxalase I bound to sepharose 4B.

Glyoxalase I bound to Sepharose 4B was used for synthesis of S-lactoyl-glutathione. The bound enzyme does not lose its activity during several months storing and can be used many times for synthesis of S-lactoyl-glutathione. This reaction product can be used as a substrate for glyoxalase II without any further purification.