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Biomedical subjects

S Javorschi

Publications and source records attributed to S Javorschi.

11 recordsLinked to original sources
PubMed2005

Two-color multiplex assay for the identification of orthopox viruses with real-time LUX- PCR.

The LUX [Light Upon eXtension] is a real-time detection system that can be used for the detection and quantification of pathogens nucleic acids. In this study we used a universal LUX approach, a variation of the LUX detection system, for identifying Orthopoxvirus nucleic acids in real time. This approach enables the design of sequence-specific primer sets in high identity genome sequences. The assay described here is designed to allow simultaneous detection of Variola and other orthopox viruses in a multiplex format, with a limit of detection in the range of 50--100 copies of the Orthopoxvirus genome. Regression analysis showed that the assay was linear over seven orders of magnitude, with 0.97 correlation coefficient. The sensitivity and specificity of the assay, as determined from a panel of 100 samples that contained nucleic acids from a variety of bacteria and viral species, were rated at 98%. Thus, the assay offers a sensitive and specific tool for simultaneous identification and quantification of Variola and other orthopox viruses, and the approach allows more flexible sequence-specific primers design for pox viruses as well as other microbial pathogens.

Animals↗
PubMed2001

Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice.

Adipocyte complement-related protein (30 kDa) (Acrp30), a secreted protein of unknown function, is exclusively expressed in differentiated adipocytes; its mRNA is decreased in obese humans and mice. Here we describe novel pharmacological properties of the protease-generated globular head domain of Acrp30 (gAcrp30). Acute treatment of mice with gAcrp30 significantly decreased the elevated levels of plasma free fatty acids caused either by administration of a high fat test meal or by i.v. injection of Intralipid. This effect of gAcrp30 was caused, at least in part, by an acute increase in fatty acid oxidation by muscle. As a result, daily administration of a very low dose of gAcrp30 to mice consuming a high-fat/sucrose diet caused profound and sustainable weight reduction without affecting food intake. Thus, gAcrp30 is a novel pharmacological compound that controls energy homeostasis and exerts its effect primarily at the peripheral level.

Adipocytes↗
PubMed2000

Rapid ELISA D-dimer testing in the exclusion of venous thromboembolism in hospitalized patients.

The clinical diagnosis of deep-vein thrombosis (DVT) and pulmonary embolism (PE) is known to be unreliable. Until now, no biological marker has been found to confirm thrombosis, but help can be gained from a biological marker ruling out the diagnosis of DVT or PE, i.e., the sensitive measurement of D-dimer (DD) species. This article summarizes our experience in introducing a rapid D-dimer test (ELISA VIDAS D-dimères test, bioMérieux) in a collaborative strategy for thrombosis diagnosis during 9 consecutive months involving 1,131 measurements. The efficacy of the DD test was very different according the type of patient, and departments where the DD test provides a real diagnostic benefit were identified. High clinical probability for thrombosis was encountered in 32 patients and radiology was carried out, although D-dimer was negative: none of these patients was found to have a thrombosis after radiologic examination. However, extensive progress must be made in test prescription to reduce the excessive rate of positive D-dimer tests (78%) and positive measurements that are not followed up by radiology (42%).

Biomarkers↗
PubMed2000

Tissue factor mRNA quantitation without prior monocyte isolation.

Monocyte tissue factor (TF) quantitation evaluates the involvement of coagulation processes in many diseases. However, technical difficulties, such as blood sampling of cells representative of the whole intravascular pool, cell isolation, protein quantitation or activity assessment, hinder reliable evaluation of TF expression by activated monocytes. Early determination of such activation can be achieved through TF mRNA quantitation by RT-PCR and sensitive product detection, such as automated electrophoresis of fluorescently labeled products. Although it is very sensitive, this method has its limitations. It needs to be standardized using other mRNA that display two main characteristics: the absence of upregulation during inflammation and similar levels of expression when compared with the target mRNA. Widely used standardization housekeeping genes such as HLA or GAPDH genes only meet the former requirement. We demonstrate here that CD11b gene expression meets both conditions. Moreover, because of its specific expression in myelomonocytic cells, it is possible to avoid further monocyte purification from a regular mononuclear cell preparation. A rapid, sensitive, specific and accurate way to evaluate monocyte TF expression is described in this paper.

Cell Separation↗
PubMed1998

D-dimer strategy in thrombosis exclusion--a gold standard study in 100 patients suspected of deep venous thrombosis or pulmonary embolism: 8 DD methods compared.

DD are now recognized as a valuable tool to screen patients suspected of deep venous thrombosis or pulmonary embolism before carrying out a gold standard radiologic examination. The newest methods available claim to be able to ascertain the absence of thrombosis, but they have yet to prove their efficiency. We compared the performances of 3 reference ELISA methods (D-DI Asserachrom Stago, D-dimer Enzygnost Behring and Dimertest GOLD EIA Agen), 5 recent rapid methods (VIDAS D-Dimer bioMérieux, Instant IA Stago, Simplired Agen, Nycocard D-dimer Nycomed and Accuclot D-Dimer Sigma Diagnostics) and two routine latex methods (Dimertest American Diagnostica and FDP-Slidex bioMérieux) in 100 patients. One of the rapid quantitative methods was demonstrated to have a level of efficiency comparable to that of ELISA methods. Finally, the cost and efficiency of different strategies were evaluated, the association of a routine latex method with the VIDAS D-Dimer bioMérieux being proven to be the most efficient.

Adult↗
PubMed1997

Mild hyperhomocysteinemia and hemostatic factors in patients with arterial vascular diseases.

Mild hyperhomocysteinemia, due to genetic or to environmental factors, is now recognized as a risk factor for premature arterial disease, including peripheral arterial occlusion, thrombotic stroke and myocardial infarction. It is defined by either an increased level of fasting homocysteine or by an increased level after loading with methionine, which is more frequently altered than the former. We studied the hemostatic parameters in 88 patients with premature arterial disease (mean age 43 +/- 11 years). We confirmed previously known hemostatic alterations described in vascular patients when compared to controls, but found that, among patients, some of these parameters were more altered in hyperhomocysteinemic patients. When fasting homocysteine was increased, higher alterations were found in factors VIIIc, von Willebrand and thombin-antithrombin complexes were more elevated. When post-methionine load homocysteine was increased, alterations in fibrinolytic parameters were more pronounced.

Adult↗
PubMed1997

Proposal for objective evaluation of the performance of various functional APC-resistance tests in genotyped patients.

The aim of the present study was to evaluate the relative performance of five screening methods for APC resistance caused by the factor V:Q506 mutation: the original method Coatest APC Resistance Chromogenix, a modified method using the same reagents but a predilution 1+4 of the plasma in a factor V deficient plasma from Stago (Stago deficient V) or from Chromogenix (V-DEF Plasma), the Coatest APC Resistance V (Chromogenix), and Accélérimat from bioMérieux. Normalization was done against a pool of normal plasmas for the methods from Chromogenix. The study included 350 subjects, 219 were genotyped (174 FV:R506R, 42 FV:Q506R, 3 FV:Q506Q) and most of them were assessed by more than one method. Uncertainty in predicting the FV genotype was evaluated by statistical analysis, which provided a way to quantitate the performance of the different diagnostic approaches. Performance of each test was evaluated by its sensitivity, specificity, R.O.C. curves, positive and negative likelihood ratios (LR), and the overall performance was determined by two parameters derived from the LR curves : the maximum LR value obtained at the crossover of the two curves, and the distance between the two curves for LR = 10. Coatest APC Resistance V and Accélérimat were proven to be the methods most able to discriminate for factor V:Q506, while normalization was not shown to improve the screening performance. The original method from Chromogenix was confirmed to undergo many influences (factor XII, PAI-1, thrombin-antithrombin complexes, antithrombin III, hematocrit). Although a very good improvement was provided by the newest methods, they were shown to be influenced by protein S and/or factor V levels in the sample plasma.

Blood Coagulation Tests↗
PubMed1996

Technical and biological conditions influencing the functional APC resistance test.

Poor anticoagulant response to APC is conveniently screened by a commercially available functional test (Coatest APC Resistance) allowing identification of APC-resistant patients. These patients may then be genotyped with respect to factor V, the Arg -> Gln mutation being the principle cause of APC resistance. However, determination of phenotype generally precedes that of genotype, and the need for an "abnormality threshold" prompted a study of inter-batch variations and the clinical conditions associated with an altered APC response. The response to APC was assessed twice in plasma from 111 patients using two of four successive kit batches. A modest but significant inter-batch variability was observed. At the same time, we also tested 130 patients with retinal venous occlusion (RVO), 28 patients with glaucoma and 24 normal volunteers. The APCaPTT/aPTT ratio was found to be lower in the presence of elevated thrombin-antithrombin complexes (r = 0.167, p < 0.02) and low blood viscosity (at high shear rate: r = 0.305, p < 0.0001) independently of any alteration in genotype.

Aged↗