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June 1, 2005
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June 1, 2005
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Herein we review all of the data from linkage by genome scanning and from association studies in essential hypertension. Genome scans have yielded loci linked to hypertension on almost every chromosome. We tabulate all of these loci to highlight the striking inconsistency. Similarly, association studies have implicated > 66 genes to date, which we also list, but virtually all have failed to show consistent replication in other settings. Nevertheless, we believe that molecular genetics should eventually find all of the major gene variants for essential hypertension. This will be a great scientific achievement and lead to new treatments. The dream, however, of using this information in clinical genetic testing could turn out to be a nightmare. Thus at present the hype surrounding genes for complex polygenic diseases like hypertension far exceeds the reality.
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June 1, 2005
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Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disease caused by loss-of-function mutations in the gene encoding thymidine phosphorylase (TP). TP deficiency alters the metabolism of the nucleosides thymidine and deoxyuridine, which, in turn, produces abnormalities of mitochondrial DNA (mtDNA) including depletion, deletions, and point mutations. MNGIE is the best characterized of the expanding number of mitochondrial disorders caused by alterations in the metabolism of nucleosides/nucleotides. Because mitochondria contain their own machinery for nucleoside and nucleotide metabolism and have physically separate nucleotide pools, it is not surprising that disorders of these pathways cause human diseases. Other diseases in this group include mtDNA depletion syndromes caused by mutations on the nuclear genes encoding the mitochondrial thymidine kinase and deoxyguanosine kinase; autosomal dominant progressive external ophthalmoplegia with multiple deletions of mtDNA due to mutations in the genes encoding the muscle-isoform of mitochondrial ADP/ATP translocator; and mitochondrial DNA depletion due to toxicities of nucleoside analogues. Mutations in the deoxynucleotide carrier, a transporter of deoxynucleoside diphosphates, have been identified as a cause of congenital microcephaly. However, alterations of mtDNA have not yet been established in this disorder. Future studies are likely to reveal additional diseases and provide further insight into this new subject.
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June 1, 2005
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Tryptophan is metabolised primarily along the kynurenine pathway, of which two components are now known to have marked effects on neurons in the central nervous system. Quinolinic acid is an agonist at the population of glutamate receptors which are sensitive to N-methyl-D-aspartate (NMDA), and kynurenic acid is an antagonist at several glutamate receptors. Consequently quinolinic acid can act as a neurotoxin while kynurenic acid is neuroprotectant. A third kynurenine, 3-hydroxykynurenine, can generate free radicals and contribute to, or exacerbate, neuronal damage. Changes in the absolute or relative concentrations of these kynurenines have been implicated in a variety of central nervous system disorders such as the AIDS-dementia complex and Huntington's disease, raising the possibility that interference with their actions or synthesis could lead to new forms of pharmacotherapy for these conditions.
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June 1, 2005
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Bronchial asthma is a complex disease characterized by airway inflammation involving a Th2-cytokine, interleukin (IL)-13. A substantial body of evidence has accumulated pointing to the pivotal role of IL-13 in the pathogenesis of bronchial asthma. The evidence is categorized as (i) analyses of mouse models, (ii) expression of these cytokines in the bronchial lesions, and (iii) genetic association of the signaling molecules of these cytokines. In addition, the molecular mechanism of the signal transduction of IL-13 has also been well characterized. We have applied microarray analyses to human bronchial epithelial cultures to search for genes regulated by IL-13 and have identified a subset of disease-relevant genes by comparison with cDNA libraries derived from normal and asthmatic bronchial biopsies. Expression of squamous cell carcinoma antigen-1 (SCCA1) and SCCA2, the cysteine and serine protease inhibitors, respectively, was the highest in the bronchial epithelial cells stimulated by IL-4 and IL-13 and was augmented in the asthmatic cDNA library. Furthermore, serum levels of SCCA were also elevated in asthmatic patients. Taken together, it was supposed that SCCA may play some role in the pathogenesis of bronchial asthma, and measuring its serum level may be relevant for diagnosing or monitoring the status of bronchial asthma.
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June 1, 2005
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Nitric oxide (NO) activates soluble guanylyl cyclase, which results in an increased synthesis of cyclic guanosine 3′,5′-cyclic monophosphate (cGMP), smooth muscle relaxation and vasodilation. The heme group in soluble guanylyl cyclase binds NO and allosterically activates the catalytic site. In addition, a second allosteric site that synergistically activates the enzyme has been reported. BAY 41-2272 was reported as an NO-independent activator of soluble guanylyl cyclase. Treatment with this compound results in anti-platelet activity, a decrease in blood pressure and an increase in survival, indicating a potential for treating cardiovascular diseases. YC-1, another NO-independent activator, activates soluble guanylyl cyclase and the activity is enhanced in the presence of NO. YC-1 relaxed tissue strips in organ bath. Consistent with its biochemical activity, YC-1 induced penile erection in a conscious rat model. Recently, we found a novel series of soluble guanylyl cyclase activators that also NO-independently activate soluble guanylyl cyclase and cause penile erection, suggesting a synergy with the endogenous NO production in vivo . Here I review the NO/cGMP signal transduction pathway and define soluble guanylyl cyclase modulators as a novel approach for the treatment of cardiovascular diseases and erectile dysfunction.
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June 1, 2005
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The practice of cardiology continues to evolve along with a better understanding of the pathophysiology of cardiovascular disease and the development of new therapeutic procedures. Consequently, new demands are being made on the in vitro diagnostics industry to improve the performance of existing cardiac markers and to develop novel markers for new cardiac disease indications. Indeed, in the last 20 years there has been a progressive increase in new laboratory tests for markers of cardiac diseases. Several highly sensitive and/or specific assays for the detection of myocardial ischemic damage as well as some immunoassays for cardiac natriuretic hormones, now considered a reliable marker of myocardial function, have become commercially available. In parallel, a growing number of some novel risk factors, which can be assessed and monitored by laboratory methods, have been added to the classical risk factors for cardiovascular disease. Finally, the recent explosion of genetic analysis may soon place at the clinical cardiologist's disposal many laboratory tests for defining the diagnosis at the molecular level, assessing new risk factors, and better targeting the pharmaceutical approaches in patients with cardiovascular disease. In the present article, after a brief description of the analytical tests included in these four groups, each group's impact on clinical cardiology is discussed in detail.
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June 1, 2005
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Preanalytical issues do not readily lend themselves to systematic teaching and the approach to teaching must be modified for each of the populations who require such education. The populations to be educated include patients, nurses, medical students and physicians. The different approaches that we have tried, and continue to use are described.
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June 1, 2005
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Allergic asthma is caused by genetic and environmental factors that interact to determine disease susceptibility and severity. Several lines of evidence suggest that the interleukin (IL)-4 gene and the IL-4-receptor α ( IL-4Rα ) gene are involved in the development of atopic diseases. We sought to determine whether two polymorphism sites in IL-4 and IL-4R α chain were associated with allergic asthma in a Chinese population. We obtained DNA and clinical data from allergic asthma patients, which were compared with those of a group of healthy control subjects. The subjects were genotyped for the IL-4 C-590T promoter polymorphism and the IL-4R α chain Q576R by polymerase chain reaction-restriction fragment length polymorphism. The results showed that the IL-4 C-590T was not associated with allergic asthma in a Chinese population. However, the IL-4R α chain 576R/R was significantly increased in allergic asthma patients compared with control subjects (χ 2 =21.16; p < 0.01), and also total plasma immunoglobulin E (IgE) level was increased in allergic asthma patients. These data suggest that the IL-4R α chain 576R/R genotypes confer genetic susceptibility to allergic asthma in Chinese.
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June 1, 2005
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Transforming growth factor β 1 (TGF-β 1 ) is involved in different physiological and pathological processes, including atherogenesis. High plasma lipoprotein(a) (Lp(a)) concentration is an established independent risk factor that may interfere with the plasmin-mediated TGF-β 1 activation. Both Lp(a) and TGF-β 1 are thought to influence the expression of cellular adhesion molecules (CAMs), also involved in the process of atherogenesis. Whereas many studies confirmed the association between high plasma Lp(a) levels and coronary artery disease (CAD), conflicting results were obtained in different studies in which the changes of TGF-β 1 and CAM concentrations in CAD patients were investigated. The aim of this case-control study was to explore the association of circulating TGF-β 1 , Lp(a) and CAMs (intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin) levels with the occurrence and severity of angiographically assessed coronary artery disease. Plasma TGF-β 1 , Lp(a), ICAM-1, VCAM-1 and E-selectin concentrations were measured in 100 patients with angiographically assessed CAD and 100 healthy blood donors matched according to age and gender. Lp(a) and TGF-β 1 were significantly higher in patients than in healthy controls (p < 0.001 and p < 0.01, respectively), but no significant correlation between the TGF-β 1 and Lp(a) values was found. The CAM concentrations obtained in CAD patients did not differ significantly as compared with the corresponding values in the controls. None of the measured parameters were influenced by the severity of CAD.
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June 1, 2005
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Inflammatory response in surgery is associated with the release of cytokines. Many cytokines are produced by macrophages; therefore surgical injuries to the liver may have great influence on the release of cytokines. Ischemia creates tissue injury and may contribute to the cytokine release. A balanced ratio of pro- and anti-inflammatory cytokines is important for appropriate immune response; excessive inflammation or hypo-responsiveness can lead to post-operative complications. To determine the magnitude of the cytokine response caused by liver surgery and to evaluate the balance of pro- and anti-inflammatory cytokines released during the operation, we measured levels of tumor necrosis factor-α (TNFα), interleukin (IL)-1β, IL-6 and IL-10 in 19 patients undergoing liver resection. The results showed a continuous rise of IL-6 and a transient elevation of IL-10. Levels of TNFα remained low; IL-1β was not detected at any sampling time. We conclude that liver surgery induces cytokine response characterized predominantly by an early appearance of IL-6 and IL-10, the elevation of IL-6 may be mainly caused by splanchnic ischemia. The IL-6/IL-10 ratio could possibly reflect the balance of pro- and anti-inflammatory cytokines in liver surgery better than the TNFα/IL-10 ratio, which can well represent inflammatory status in sepsis.
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June 1, 2005
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The main objective of the present study was to analyze some aspects of the function and structure of erythrocytes with respect to hemodialysis membrane biocompatibility throughout the erythrocyte and serum antioxidant levels. The study included 36 hemodialysis patients (14 female and 22 male, age 22–79 years, median 55) treated at the Department of Nephrology and Dialysis, Split Clinical Hospital in Split, and 30 control subjects matched for age and sex. Hemodialysis was performed three times a week for 4 hours with cellulose diacetate (n = 17; 6 females and 11 males) or polysulfone (n = 19; 8 females and 11 males) membranes. The aim of the study was to assess the level of oxidative stress in these patients by measuring catalytic concentrations of superoxide dismutase and glutathione peroxidase in erythrocyte lysate and scavenger systems related to free hemoglobin in serum (haptoglobin, hemopexin and bilirubin). In comparison with control values, the mean catalytic concentrations of superoxide dismutase were increased and catalytic concentrations of glutathione peroxidase decreased in patients before hemodialysis irrespective of the membrane used. Immediately after hemodialysis with either membrane, the mean catalytic concentrations of superoxide dismutase returned to the control range, while those of glutathione peroxidase were still decreased compared to control values, without any significant difference between the cellulose diacetate and polysulfone membranes. The predialysis and postdialysis values of haptoglobin, hemopexin and bilirubin in patient sera were within the range of control values. Comparison of the cellulose diacetate and polysulfone membranes showed no significant differences in the erythrocyte content of antioxidant enzymes and the scavenger system related to free hemoglobin in serum before and after hemodialysis.
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June 1, 2005
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The rationale of this study was to understand the complexity of kinetics of fluorogenic phospholipid substratesas well as contradictory findings of clinical papers measuring phospholipase A 2 (PLA 2 ) activity using different methodologies. The aim was to recommend to clinicians and researchers what substrate in conjunction with what assay should be used. Two methods, (i) continuous fluorometric assay and (ii) high performance thin layer chromatography (HPTLC) on microplates combined with quantitative image scanning, were studied with three different substrates (bis-BODIPY ® FL C11-PC, NBDC 6 -HPC ® , PED6 ® ). The study demonstrates that NBD-PC is not a suitable substrate to measure PLA 2 activity using a spectrofluorometer. On the other hand, NBD-PC gives the highest and most reproducible integrated light intensities (ILIs) in HPTLC studies. Slow time-dependent increases in fluorescence intensities recorded with biological samples in fluorometers, but not caused by substrate splitting, had to be classified as “perturbation kinetics”. PLA 2 activities in blood samples of 26 unmedicated schizophrenia patients and 26 age-matched healthy controls were measured by the spectrofluorometric method and then compared with the activity data obtained with the HPTLC method. A significant group difference was found only with the HPTLC. In order to get more reliable results, we recommend that clinicians and researchers use NBD-phosphatidylcholines as PLA 2 substrates in biological samples and start with an analytical separation of reaction products followed by image analysis of the fluorescent spots.
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June 1, 2005
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In this paper a method for the simultaneous quantification of the anti-fungal drug itraconazole and its co-active metabolite hydroxyitraconazole in plasma employing liquid chromatography tandem-mass spectrometry and automated solid-phase extraction is described. The method proved rugged, enables short turn-around times and is highly specific. Since there is growing evidence for the importance of therapeutic drug monitoring of itraconazole in the prophylaxis and treatment of invasive fungal infections, the method described here is of interest for a large number of tertiary care hospital laboratories.
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June 1, 2005
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Sirolimus appears as a new potent immunosuppressive agent taking advantage of therapeutic drug monitoring to optimize its use in organ transplantation. In the absence of any available commercial immunoassay it was mandatory to develop chromatographic assays. Some methods have already been proposed to quantify sirolimus in whole blood, based either on HPLC-UV, liquid chromatography-mass spectrometry (LC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS). We have developed a new faster and simpler LC-MS/MS method to quantify sirolimus in blood using ascomycin as an internal standard and multiple reaction monitoring (MRM) acquisition mode. This method displays a limit of detection of 0.3 μg/l, and the intra-assay reproducibility ranges from 4.1–7.9%. The pre-analytical preparation steps are quite similar to those required for semi-automated immunoassays. Ascomycin and sirolimus present retention times of 0.89 and 0.93 min, respectively, and the turnaround time for a result (2.5 min) is also similar to that observed using a clinical analyzer. The comparison performed between HPLC-UV and LC-MS/MS displays good correlation (r = 0.949). The LC-MS/MS method described above has been used routinely for more than 2000 patient blood specimens and may present several advantages over existing methods, e.g. , specificity with sufficient sensitivity, rapidity, and small blood sampling (10 μl), making it particularly adapted for routine clinical use.
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June 1, 2005
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The formation of superoxide partially accounts for the well-known oxygen enhancement of radiation-induced biochemical changes and cell damage. Radioprotective effects of copper (II), manganese (IV) or vanadium (IV) complexes, of superoxide dismutasemimetic activity, on body weight, survival rate and some biochemical parameters in pre-treated irradiated, untreated irradiated and treated non-irradiated female albino rats have been studied 24 h after whole body γ-irradiation at a dose level of 6 Gy. Survival time, body weight, red blood cell (RBC) and white blood cell (WBC) counts, hemoglobin (Hb) concentration, percentage of hematocrit (Hct%), reduced glutathione (GSH), serum total protein, albumin, globulin (G), blood urea, creatinine and cholesterol were estimated, as well as the activities of blood superoxide dismutase (SOD), glutamate-oxaloacetic (GOT) and glutamate-pyruvic (GPT) transaminases, and alkaline phosphatase were assessed. A significant decline was shown in body weight, survival rate, the mean values of RBC and WBC counts, Hb and Hct percentages, and GSH concentration, as well as blood SOD activity, in whole body γ-irradiated rats compared with the control non-irradiated rat group. The mean activity values of alkaline phosphatase, GOT and GPT, as well as the average values of blood urea, creatinine, total cholesterol, total protein and globulin were significantly elevated, while the average values of albumin and the albumin/globulin ratio were decreased in γ-irradiated rats compared with the corresponding values of the normal control rat group. Pre-treatment of rats with either manganese or vanadium complexes resulted in a significant increase in survival rate and body weight over that of the non-treated irradiated rat group. Pretreatment of rats with copper (II), manganese (IV) or vanadium (IV) complexes caused a significant increase in RBC and WBC counts, Hb concentration, HCt (%), GSH content and SOD activity in blood when compared to the irradiated rat group without treatment. The administration of copper (II), manganese (IV) or vanadium (IV) complexes prior to irradiation exposure resulted in a significant decrease in GOT and GPT activities in addition to blood urea, creatinine, cholesterol, globulin and total protein contents, while each complex exhibited a significant increase in plasma alkaline phosphatase, albumin, and the albumin/globulin ratio compared to the untreated irradiated rat group. Administration of vanadium (IV), manganese (IV) or copper (II) complexes in non-irradiated rats caused a significant increase in SOD activity without changing other biochemical parameters compared with the corresponding values of the normal control rat group. We conclude that these metallo-elements, particularly manganese (IV) and vanadium (IV) complexes of 2-methylaminopyridine, have radiation protection and radiation recovery. Furthermore, these metal complexes offer a new approach to overcome the pathological effects of ionizing radiation and suggest their use as a physiological approach to preventing or perhaps predominantly facilitating recovery from radiation injury.
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June 1, 2005
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The recent increase in the elderly population, current health trends and awareness of age-related changes in the male endocrine system, have led to discussions about the role of the hormonal changes in the aging process in males. Better prevention and treatment of suboptimal health status and age-related diseases in aging men are based on an improved understanding of aging, particularly of the significance of age-associated hormonal changes. The aims of this study were 1) to evaluate the age dependence of the serum concentrations of the following important hormonal parameters in adult males using the IMMULITE ® 1 automated assay system (DPC, Los Angeles): testosterone, dehydro-epiandrosterone sulfate (DHEAS), estradiol (E2), sex hormone binding globulin (SHBG), lutropin (LH), follitropin (FSH), cortisol, prolactin, thyroid stimulating hormone (TSH), free triiodothyronine (fT3), free thyroxine (fT4) and the growth hormone-dependent parameters insulin-like growth factor (IGF-I) and IGF-binding protein-3 (IGFBP-3) and 2) to derive the following parameters: calculated free testosterone (cFT), ratio of calculated free testosterone to total testosterone (% cFT) and free androgen index (FAI). We found a significant decrease between the 1–30-year age group and the >70-year agegroup for total testosterone (−42.4%), FAI (−65.5%), cFT (−60.0%), % cFT (−30.0%), DHEAS (−71.9%), E2 (−35.4%), TSH (−23.6%), IGF-I (−40.3%) and IGFBP-3 (−26.5%). Since the decreases in the FAI and cFT were greater than that of total testosterone and because these derived parameters reflect the biologically active fraction of testosterone, FAI and cFT are better markers for androgen deficiency in males. In contrast, a significant increase with age was observed for SHBG (+61.2%73), LH (+40.0%), FSH (+98.3%) and cortisol (+54.2%). No significant alterations with age were observed for prolactin, fT3 and fT4. The study demonstrates that determining complete profiles of the androgenic, gonadotropic, adrenocortical, thyroid, pituitary and growth hormone/IGF endocrine axes in middle-aged and elderly men may be helpful in obtaining a correct clinical diagnosis for various hormonal disorders.
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June 1, 2005
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Over the past decade, numerous papers have addressed the various methodological problems encountered with free thyroxine (FT 4 ) assays. We evaluated the clinical performance of nine FT 4 assays in five centres, using a panel of 310 sera: 156 from euthyroid controls; 27 from hyperthyroid patients; 34 from untreated hypothyroidism; 22 from patients with renal failure; 30 from women in the last trimester of pregnancy; 23 from patients on thyroid substitutive therapy; and 18 from patients treated with amiodarone. Only three methods showed a Gaussian distribution of FT 4 concentrations. Reference ranges were calculated using the 2.5th and 97.5th percentiles. A significant difference was observed between FT 4 values in men and women. The areas under the receiver operating characteristic (ROC) curves ranged from 0.996 to 1 for hyperthyroidism and from 0.973 to 1 for hypothyroidism. In sera from patients with renal failure and from pregnant women, method-dependent biases were observed and confirmed with dilution experiments. In conclusion, current FT 4 assays show good performance regarding the diagnosis of overt dysthyroidism. Nevertheless, FT 4 measurements are still vulnerable to method-dependent artefacts in particular populations such as patients with renal failure and pregnant women.
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June 1, 2005
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Determination of C-reactive protein (CRP) and high-sensitivity CRP (hs-CRP), markers of systemic inflammation and atherosclerosis risk, usually requires serum samples, while heparin plasma samples are routinely collected for routine chemistry. This study evaluates the use of heparin plasma samples for hs-CRP and CRP determinations using immunoturbidimetric methods on an Olympus system. As a result, analytical performance, including linearity, imprecision values, and determination from 185 clinical samples, are not affected by the type of sampling, validating the use of heparin plasma samples for CRP and hs-CRP determination with Olympus reagents.
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June 1, 2005
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The comparability between glucose concentrations measured in various sample systems is still a matter of debate. Decision limits are usually determined in venous plasma and then converted to either blood or to the aqueous compartment (activity). The conversion factors recommended have not yet been generally accepted. In the present study, glucose concentrations were determined in blood and plasma with an Ebio analyzer (molarity) and in the aqueous compartment with both an EML 105 and an Omni (molality). All analytical results were referred to the same aqueous standard solution. The Ebio results agreed with reference method values in control materials. Concentrations determined in the various sample systems from patients (molarity) correlated well with the molality values measured either with the EML or the Omni. However, the mean values of the EML were not consistent with those derived theoretically by considering the different water content. With the Omni, only molality values in whole blood appeared plausible, but not in plasma, although the two sample systems should provide identical molality values. The EML results were almost identical in whole blood and plasma. Theoretically, glucose molality would be the ideal diagnostic quantity. However, no diagnostic advantage of molality determined in whole blood with the Omni vs . molarity values could be detected in a group of 40 non-diabetic and 27 diabetic subjects.
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June 1, 2005
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The performance characteristics and diagnostic accuracy of a new rapid automated quantitative immunoturbidimetric D-dimer assay, Auto-Dimer (Biopool, Umeå, Sweden) were evaluated in a population of 135 outpatients with suspected deep-vein thrombosis of lower limb. Enzyme-linked immunosorbent assay was used as the reference method. The Auto-Dimer assay showed good reproducibility. The correlation between Auto-Dimer and enzyme-linked immunosorbent assay was high (r = 0.91, p < 0.05) and agreement in classifying patients above or below the cut-off was good (κ coefficient 0.74, 95 % CI 0.62–0.86). At a cut-off of 340 μg/l the sensitivity and specificity of Auto-Dimer were high (100% and 61%, respectively). These data show that Auto-Dimer is a reliable screening test for exclusion of deep-vein thrombosis. The assay could be included in prospective patient management studies in order to obtain further information on its clinical usefulness.
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June 1, 2005
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It is a common belief that laboratory investigation processes were developed after the 16th century and that before that time no attempts were made to attain a diagnosis by investigating material coming from the human body. In this paper we present data extracted from Byzantine codices that support the following thesis: The idea of examining human excrement for diagnostic purposes has its roots in the Roman and Byzantine eras. The lack of technological means was no obstacle for the doctor to create an “examinational” mind, i.e. , to try to correlate the macroscopic findings in the excrement with the pathophysiological mechanism that induced it, using only the human senses.
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June 1, 2005
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June 1, 2005
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July 27, 2005
