Mutations in the basic core promoter region of hepatitis B virus. Relationship with precore variants and HBV genotypes in a Spanish population of HBV carriers.

Background/aims: To determine the prevalence and significance of hepatitis B virus (HBV) basic core promoter (BCP) mutations and to establish their relationship with precore (preC) mutations, HBV genotypes and HBV-DNA levels.

Methods: BCP and preC mutations and genotypes were determined by sequencing.

Results: Genomic analysis was performed in 129 (71%) of 182 patients.

Application of receiver operating characteristics curve (ROC) analysis when definitive and suspect morphologic flags appear in the new Coulter LH 750 analyzer.

Workflow efficiency is important in every laboratory. Manual assessment of white blood cell counts and differentials that have been rejected by an automated analyzer is one of the most time-consuming tasks in the routine hematology laboratory. In this study, receiver operating characteristics (ROC) curve analysis was used for the first time when anomalous distribution and suspect flag alarms appeared in hemograms carried out with the new Beckman Coulter LH 750 analyzer.

Use of the Advia 120 hematology analyzer in the differential cytologic analysis of biological fluids (cerebrospinal, peritoneal, pleural, pericardial, synovial, and others).

The centralization of our laboratories and the demand for new parameters to measure have led to an increase in the number of biological fluid samples, which are generally sent for urgent analysis. Due to this they cannot be processed by manual methods. Meeting this increased demand for assistance is a challenge for the laboratory, and the challenge has been met by the automated hematology area.

Validation of the Coulter LH 750 in a hospital reference laboratory.

Validation of the Coulter LH 750 was carried out in our central hospital laboratory, which processes 1500 hemograms per day for patients admitted into the 4 hospitals of our hospital complex and their corresponding outpatient departments. It is the reference laboratory for the provincial health care area. The analytical quality and the practical use of the instrument were studied, and we obtained within- and between-run imprecision estimates with reference controls of between 0.

Detection of polymorphisms at exons 3 (Tyr113-->His) and 4 (His139-->Arg) of the microsomal epoxide hydrolase gene using fluorescence PCR method combined with melting curves analysis.

An association between exon 3 polymorphisms of the gene encoding microsomal epoxide hydrolase (mEH) and susceptibility to the development of chronic obstructive pulmonary disease (COPD) has been described. We have developed two methods for detecting polymorphisms at exons 3 (Tyr113-->His) and 4 (His139-->Arg) of the mEH gene based on different melting temperatures (T(m)) of fluorescent-labeled oligonucleotide hybridization probes using single-step assays that combine fluorescence PCR and melting curve analysis (LightCycler methodology). DNA was extracted from blood in 79 COPD patients and 146 healthy controls.

Rapid screening for alpha1-antitrypsin deficiency in patients with chronic obstructive pulmonary disease using dried blood specimens.

We describe two reliable methods for high-throughput screening of proteinase inhibitor (PI) S and PI Z alpha(1)-antitrypsin (alpha(1)-AT) deficiency alleles from dried blood spot (DBS) specimens using the LightCycler fluorimetric analyzer. The method was used to study 72 patients with chronic obstructive pulmonary disease. Results were confirmed with DNA sequencing.

Quantitative detection of hepatitis B virus DNA in serum by a new rapid real-time fluorescence PCR assay.

A sensitive and accurate HBV DNA quantification assay is essential for monitoring hepatitis B virus (HBV) replication. This study evaluated a real-time PCR method performed in the LightCycler analyser for quantitative HBV DNA assay. HBV DNA results with this method were compared with those obtained using a branched-chain DNA (bDNA) solution hybridization assay.

Role of hepatitis B, C, and D viruses in dual and triple infection: influence of viral genotypes and hepatitis B precore and basal core promoter mutations on viral replicative interference.

The interactions among hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatitis delta virus (HDV) were studied by measuring HBV-DNA and HCV-RNA levels and by determining the influence of viral genotypes and mutations in HBV basal core promoter (BCP) and precore regions. We included 65 consecutive patients, 25 HBV/HCV, 18 HBV/HDV, and 22 HBV/HCV/HDV. Controls consisted of 55 patients with chronic HBV and 55 with chronic HCV infection.

Are equally spaced specimen collections necessary to assess biological variation? Evidence from renal transplant recipients.

The established method for determining the components of biological variation (BV) requires equispaced time intervals between samplings. In a previous study, we determined BV in renal post transplantation patients, taking advantage of the samples obtained within their clinical treatment protocol (not necessarily equispaced). To confirm the validity of this practice, we sought to determine if the use of varying sampling intervals has an effect on the results obtained in such biological variation studies.

External quality assessment of urine analysis in Europe. Results of a round table discussion during the symposium 'From uroscopy to molecular analysis', Seeon, Germany, September 18-20, 1999.

Interlaboratory surveys on urine quantities have only recently been introduced in several European countries. Representatives of 10 European countries exchanged their experiences during an international urinalysis meeting held in September 1999. Although still not mandatory in most areas, more than 5000 laboratories participated in external quality assessment programs in the countries represented.

European multicentre evaluation of the Super Aution SA-4220 urinalysis analyser.

A multicentre evaluation of the urine test strip analyser Super Aution-4220 was carried out in six laboratories. The analytical performance of the instrument with regard to imprecision, linearity, detection limit, drift, carry-over and method comparison was studied. Using the Aution stick 8 test strip the pH, glucose, protein, ketones, bilirubin, blood, urobilinogen and leukocyte esterase were analysed.

Fosinopril ameliorates exogenous cholesterol-induced incipient glomerular lesions in obese Zucker rats. Effects on eicosanoid secretion.

Background: To date, the role of dietary cholesterol as a risk factor for some diabetic nephrophathy, such as mesangial expansion and glomerular lesions, is unknown. Controversy also exists regarding the effects of prostaglandin-induced changes in glomerular haemodynamics on the appearance of glomerulosclerosis.

Methods: We have used obese Zucker rats (OZRs) as a model of early nephrophathy to evaluate the effect of hypercholesterolaemic diet on glomerular prostaglandin secretion and on the development of glomerular lesions.

[Interference in blood coagulation tests on lipemic plasma. Correction using n-hexane clearing].

Purpose: To eliminate the bias in the plasmatic coagulation test caused by lipemic samples by means of with addition of n-hexane in the ACL-3000 analyzer (Instrumentation Laboratory).

Material And Methods: The turbidity caused by lipemia was simulated by addition of increasing quantities of Intralipid 20% (interferent) to samples from patients with normal pathological coagulation tests (with oral anticoagulant therapy). We compared the results of PT (prothrombin time), fibrinogen (derived) and APTT (activated partial thromboplastin time) before and after the addition of n-hexane.

Potential interfering substances on Falcor-600 and Dax-48 analytical systems.

The increasing availability and use of automatic analysers in clinical chemistry have revealed a number of endogenous interferences. We evaluated the effect of bilirubin, haemolysis and lipaemia on the Falcor-600 analytical system (Menarini Diagnostics) and the Dax-48 (Bayer Diagnostic). We studied the potential endogenous interferences in the measurement of serum glucose, urea, creatinine, cholesterol, triacylglycerols, total bilirubin, total protein, aspartate aminotransferase, alanine aminotransferase and gamma-glutamyltransferase on both analysers; and albumin, direct bilirubin, uric acid, inorganic phosphorus, iron, calcium, magnesium, chloride, sodium, potassium, alkaline phosphatase, amylase, lactate dehydrogenase and creatine kinase on the Dax-48.

Effects of long-term therapy with either heparin or low-molecular-weight heparin on serum lipid levels. A prospective study.

Beside the anticoagulant and antithrombotic activity, heparin also exerts a lipolytic activity. In a prospective study on patients with venous thromboembolism and some contraindications to coumarin therapy, a low-molecular-weight heparin (Fragmin) was compared to unfractioned (UF) heparin in terms of both efficacy and safety. A secondary aim was to study the influence of both types of heparin on serum lipid levels.

Peritoneal transport evaluation in peritonitis: comparison between methods.

Peritoneal dialysis patients may need solute permeability transport evaluation during acute peritonitis. The aim of this study was to assess if the simplified mass transfer coefficient (MTCS) or the peritoneal equilibration test (PET) was equivalent to the complex MTC (MTCX) in solute transport evaluation during acute peritonitis in continuous ambulatory peritoneal dialysis (CAPD) patients. We studied 15 episodes of peritonitis (PTIS).

Criteria for the assessment of analyser practicability.

This article lists the theoretical criteria that need to be considered to assess the practicability of an automatic analyser. Two essential sets of criteria should be taken into account when selecting an automatic analyser: 'reliability' and 'practicability'. Practibility covers the features that provide information about the suitability of an analyser for specific working conditions.

Evaluation of the IL-Phoenix chemistry electrolyte analyser.

This paper reports an evaluation of the IL-Phoenix Chemistry/Electrolyte Analyser; the evaluation was carried out in accordance with internationally recognized guidelines. The evaluation was performed in three steps: evaluation in routine conditions; assessment of interferences; and study of practicability. Seven constituents were studied under routine working conditions.

Mass transfer coefficient: comparison between methods.

The mass transfer area coefficient (MTC) is the best parameter for solute transport evaluation in continuous ambulatory peritoneal dialysis (CAPD) patients. We compared three simplified MTC methods (calculated according Garred, Krediet, or Lindholm) and the peritoneal equilibration test (PET) (Twardowski) to complex MTC (MTCX) (Randerson and Farrell) for urea and creatinine, by means of 29 tests performed in 24 stable CAPD patients. There were no significant differences (paired t-test) between MTCX and each of the simplified MTC, except for creatinine MTC calculated by Krediet's method, which was significantly different (MTCX: 9.

Peritoneal function tests: usefulness of simplified methods.

The Peritoneal Equilibration Test (PET) as standardized by Twardowski (T) and simplified Mass Transfer Coefficients (MTC) as per Garred (G) and Krediet (K) have been employed in our patients to assess: 1) the reproducibility of results, 2) correlation between methods, and 3) usefulness of patient categorization by simplified methods in contrast to PET patterns. We have performed 29 standardized PET (Dianeal 2.5%) in 24 stable CAPD patients.

Evaluation of the Chem 1 analyzer.

The selective multitest Technicon Chem 1 analyser was evaluated according to the guidelines of the 'Comisión de Instrumentación de la Sociedad Española de Química Clínica', and the protocols of the 'European Committee for Clinical Laboratory Standards' and 'Commission de validation de techniques' of the 'Société Française de Biologie Clinique'. The evaluation was performed in three steps: evaluation in routine conditions, assessment of the interferences and study of practicability. Under routine working conditions, eighteen constituents were studied.

Evaluation of the Dacos 3.0 analyser.

The selective multitest Coulter Dacos 3.0 analyser was evaluated according to the guidelines of the Comisión de Instrumentación de la Sociedad Española de Química Clínica and of the European Committee for Clinical Laboratory Standards.THE EVALUATION WAS PERFORMED IN FOUR STEPS: examination of the analytical units; evaluation of routine working; study of interferences; and assessment of practicability.