Carvedilol Inhibits Matrix Metalloproteinase-2 Activation in Experimental Autoimmune Myocarditis: Possibilities of Cardioprotective Application.

AIMS: Acute myocarditis is a potentially lethal inflammatory heart disease that frequently precedes the development of dilated cardiomyopathy and subsequent heart failure. At present, there is no effective standardized therapy for acute myocarditis, besides the optimal care of heart failure and arrhythmias in accordance with evidence-based guidelines and specific etiology-driven therapy for infectious myocarditis. Carvedilol has been shown to be cardioprotective by reducing cardiac pro-inflammatory cytokines present in oxidative stress in certain heart diseases. However, effects of carvedilol administration in acute myocarditis with its impact on matrix metalloproteinases' (MMPs) activation have not been elucidated. METHODS AND RESULTS: Carvedilol in 3 doses (2, 10, and 30 mg/kg) was given daily to 3 study groups of rats (n = 8) with experimental autoimmune myocarditis by gastric gavage for 3 weeks. In comparison to untreated rats (n = 8) with induced myocarditis, carvedilol significantly prevented the left ventricle enlargement and/or systolic dysfunction depending on the dose in study groups. Performed zymography showed enhanced MMP-2 activity in untreated rats, while carvedilol administration reduced alterations. This was accompanied by prevention of troponin I release and myofilaments degradation in cardiac muscle tissue. Additionally, severe inflammatory cell infiltration was detected in the nontreated group. Carvedilol in all doses tested, had no impact on severity of inflammation. The severity of inflammation did not differ between study groups and in relation to the untreated group. CONCLUSIONS: The protective effects of carvedilol on heart function observed in the acute phase of experimental autoimmune myocarditis seem to be associated with its ability to decrease MMP-2 activity and subsequently prevent degradation of myofilaments and release of troponin I while not related to suppression of inflammation.

Diagnostic usefulness of sCD163, procalcitonin and neopterin for sepsis risk assessment in critically ill patients.

BACKGROUND: Sepsis is one of the most common causes of hospitalization and it is characterized by a high mortality rate in spite of the great progress in diagnosis and treatment achieved in recent years. Early diagnosis of sepsis is one of the most important elements of effective treatment. The clinical symptoms are not specific and biomarkers are considered to be useful tools in sepsis diagnostics. OBJECTIVES: The aim of our study was to evaluate the diagnostic value of sCD163 as a marker of sepsis and a comparison of it with procalcitonin and neopterin in ICU patients. MATERIAL AND METHODS: Concentrations of PCT, sCD163 and NPT were measured in 52 serum samples collected from 30 patients of the Department of Anesthesiology and Intensive Therapy of the University Hospital in Wroclaw. Venous blood was collected on the 1st and 3rd day of hospitalization. The Human CD163 Quantikine ELISA Kit was used to determine the concentrations of sCD163. Neopterin concentrations were measured by a Neopterin ELISA kit. PCT was measured at the University Center of Laboratory Diagnostics in Wroclaw using an automatic VIDAS® B.R.A.H.M.S. PCT assay. RESULTS: Our study showed that there was a significant difference between the values obtained in the study and the reference group for PCT (p < 0.0001), sCD163 (p = 0.0001) and NPT (p = 0.0001), whereas there was no difference observed between the samples obtained on the 1st and 3rd day (p = 0.5129). The area under the ROC curve was 0.847, and was comparable to the AUC of procalcitonin (0.840), and slightly higher than the AUC of neopterin (0.763), although these differences were not significant (p = 0.2990 and p = 0.9329, respectively). CONCLUSIONS: sCD163 and neopterin are promising parameters in the diagnosis of sepsis, and their value in the diagnosis of sepsis in critically ill patients may be comparable to procalcitonin.

An efficient method for isolation of representative and contamination-free population of blood platelets for proteomic studies.

To date, there has been no ideal method for blood platelet isolation which allows one to obtain a preparation devoid of contaminations, reflecting the activation status and morphological features of circulating platelets. To address these requirements, we have developed a method which combines the continuous density gradient centrifugation with washing from PGI2-supplemented platelet-rich plasma (PRP). We have assessed the degree of erythrocyte and leukocyte contamination, recovery of platelets, morphological features, activation status, and reactivity of isolated platelets. Using our protocol, we were able to get a preparation free from contaminations, representing well the platelet population prior to the isolation in terms of size and activity. Besides this, we have obtained approximately 2 times more platelets from the same volume of blood compared to the most widely used method. From 10 ml of whole citrated blood we were able to get on average 2.7 mg of platelet-derived protein. The method of platelet isolation presented in this paper can be successfully applied to tests requiring very pure platelets, reflecting the circulating platelet state, from a small volume of blood.

[Diagnostic utility of RHD-gene detection in maternal plasma in the prophylaxis of feto-maternal Rh-incompatibility]. / Diagnostyczna przydatnosc detekcji genu RHD w osoczu ciezarnych w profilaktyce konfliktu matczyno-plodowego na tle antygenu D z ukladu Rh.

OBJECTIVES: The aim of the study was analytical validation of prenatal noninvasive fetal RHD detection in maternal plasma and the preliminary assessment of its clinical utility Introduction of this noninvasive test into routine diagnostic use is important for more rational and safe immunoprophylaxis. MATERIAL AND METHODS: RHD gene was detected using real-time PCR. Primers and probes complementary to sequence on exons 7 and 10 were chosen. Samples with RHD-negative results were examined with additional tests to confirm the proper isolation of cell-free fetal DNA (cffDNA). For male fetuses, the presence of fetal DNA was confirmed by detection of the male genetic marker (SRY gene) using Quantifiler Duo kit. In the case of SRY-negative result we used mini SGM test, which is based on the detection of short-tandem repeat polymorphism differences between maternal and fetal DNA. RESULTS: Diagnostic accuracy of RHD test was 96.82%, while diagnostic value of SRY determination was lower (87.50%). Mini SGM test was able to confirm the presence of fetal DNA in 77% of the cases. CONCLUSIONS: Effectiveness of the proposed procedure of prenatal noninvasive RHD determination and cffDNA confirmation is high, on condition proper control samples and suitable verifying tests are used.