[Transitional cell carcinoma of the bladder. Evaluation of plasma levels of cellular fibronectin as a stage-dependent marker]. / Das Transitionalzellkarzinom der Harnblase. Evaluation von zellulärem Fibronektin im Plasma als stadienabhängiger Marker.

Up to now markers for transitional cell carcinoma of the bladder (TCC) are missing. Fibronectin (FN) seems to play a key role in progression and invasion of malignant tumors. The aim of this study was to assess the value of cellular FN (cFN), a more specific subform of produced FN, in different stages of TCC.cFN was determined using a highly sensitive immunoassay which we developed. Blood samples were taken of 45 patients with the first diagnosis of TCC before undergoing TUR-B and 6 patients with metastatic TCC before chemotherapy; 70 patients with nonmalignant urological disorders served as a control group.Patients with TCC showed significantly elevated cFN plasma levels compared to controls (p<0.05). Patients with muscle-invasive disease (n=15) showed significantly higher cFN plasma levels compared to the group with superficial TCC. Patients with metastatic TCC showed the highest, but not significantly elevated cFN plasma levels compared to patients with muscle-invasive TCC.The elevated cFN plasma levels in TCC underline the important role of cFN for tumor progression and its potential role as a marker for TCC. Upcoming investigations are necessary to prove the value of the potential marker cFN during follow-up and its impact as a prognostic factor for recurrence and progression of TCC.

Isolated, well-defined organovanadium(iii) on silica: single-site catalyst for hydrogenation of alkenes and alkynes.

A well-defined, isolated, single-site organovanadium(iii) catalyst on SiO2 [(SiO2)V(Mes)(THF)] was synthesized via surface organometallic chemistry, and fully characterized using a combination of analytical and spectroscopic techniques (EA, ICP, 1H NMR, TGA-MS, EPR, XPS, DR-UV/Vis, UV-Raman, DRIFTS, XAS). The catalyst exhibits unprecedented reactivity in liquid- and gas-phase alkene/alkyne hydrogenation. Kinetic poisoning experiments revealed that 100% of the V sites are active for hydrogenation.

Singlet oxygen ((1)O2) inactivates plasmatic free and complexed alpha2-macroglobulin.

alpha2-macroglobulin (alpha2M) is a broad-spectrum proteinase inhibitor and one of the major plasma proteins in humans. Activated phagocytes (especially granulocytes) generate large amounts of oxidants of the HOCI- and chloramine-type that release the mild nonradical, excited (light-emitting) oxidant singlet oxygen ((1)O2). These oxidants have been shown to inactivate several specific serine protease inhibitors in human blood [e.g., alpha1-antitrypsin or alpha2-antiplasmin (plasmin inhibitor)]. The studies reported here demonstrate that nonradical oxidants also inactivate plasmatic alpha2M. The effective dose for 50% inactivation (ED50) of plasmatic alpha2M is similar to that for plasmatic alpha2-antiplasmin. Chloramines are about 1,000-fold more effective than hydrogen peroxide (ED50)=0.75 micromol chloramine T/50 microl plasma). Serine protease-serine protease inhibitor complexes are resistant to oxidants. In contrast, here it is shown that alpha2-macroglobulin, even after binding to serine proteases is sensitive to oxidation, the captured protease is released from the protease/alpha2M complex. This is the first time that oxidative inactivation of a complexed (i.e., bound to a target protease) human protease inhibitor has be shown. The (1)O2 inhibitors methionine, cysteine, cystine, or ascorbate-in contrast to the oxy-radical scavengers mannitol, superoxide dismutase, or catalase-antagonize the chloramine/NaOCl-mediated inactivation of both uncomplexed and complexed alpha2M. Thus, the oxidant involved here is of nonradicalic nature and has reaction characteristics of (1)O2. For the inhibitory function, critical oxidizable methionines or the internal thiol-ester might be targets for (1)O2. Consequently, alpha2M can also be considered a carrier for proteases, since the alpha2M-complexed proteases regain full activity in an oxidative environment. In local areas of inflammation or thrombolysis, activated phagocytes could create microenvironments of uncontrolled protease activity by generation of (1)O2.

Evaluation of serum aminoterminal procollagen type III propeptide as an index of portal hypertension and esophageal varices in chronic liver diseases.

The concentration of the aminoterminal propeptide of type III procollagen (P-III-P) was determined in serum of cubital vein and hepatic vein of patients with various types of chronic liver diseases (n = 111) and correlated with the portal venous pressure and with the degree of esophageal varices. The P-III-P level in all chronic liver diseases was correlated (rS 0.542, p less than 0.001) with the portal venous pressure, but in liver fibrotic subjects (n = 29) this correlation (rS 0.310) was not significant, in liver cirrhosis (n = 30) the respective correlation was found to be weak (rS 0.333, p less than 0.05) and similar to that in patients with unspecified chronic liver diseases (n = 52) (rS 0.425, p less than 0.01). Sensitivity and specificity of P-III-P at a cut-off concentration of 12 ng/ml for portal hypertension (portal vein pressure 5 mm Hg) are 0.93 and 0.42, respectively, the diagnostic efficiency is 0.67. Predictive values at the same cut-off level of P-III-P and an assumed prevalence of portal hypertension of 50% are 0.62 and 0.85 for the positive and negative test result, respectively. The level of P-III-P is not related to the degree of esophageal varices. The mean P-III-P concentration in the hepatic vein was found to be significantly (p less than 0.001) higher (about 35%) than that in the cubital vein. It is concluded that P-III-P is not an useful parameter for diagnosis of portal hypertension and monitoring of portal vein pressure and of the degree of esophageal varices.

The predictive value of serum laminin for portal hypertension in chronic liver diseases.

The concentration of laminin, a high molecular weight basement membrane glycoprotein, was determined with a competitive radioimmunoassay in serum from the hepatic and cubital veins of patients with chronic liver diseases (n = 175), and correlated with portal venous pressure calculated from the hepatic vein pressure gradient. The level of laminin in the hepatic vein (mean value: 1.83 U/ml) was significantly (p less than 0.05) higher than that in the cubital vein (mean value 1.68 U/ml). In both vascular regions the glycoprotein levels increased with the degree of fibrosis, reaching their highest concentrations in cirrhosis (2.16 +/- 0.84 U/ml, p less than 0.001) (normal range: 0.81-1.43 U/ml). In all chronic liver diseases there was a significant positive correlation between the level of serum laminin and portal vein pressure (rs 0.70, p less than 0.001), which prompted us to calculate some criteria of the diagnostic validity of raised laminin for portal hypertension (portal venous pressure greater than or equal to 5 mm Hg). At a cut-off concentration of laminin of 1.45 U/ml, sensitivity is 0.87, specificity 0.74, diagnostic efficiency 0.81, and the likelihood ratio 3.4. Positive and negative predictive values at the same cut-off and at a prevalence of portal hypertension in this study of 50% are 0.77 and 0.85, respectively. Serum laminin may prove to be a potentially useful biochemical marker of portal hypertension.

Alternative medicine. Achieving balance between herbal remedies and medical therapy.

The case patient was taking multiple herbal preparations as well as the prescription hypnotic zolpidem. The combination was probably increasing the patient's confusion, agitation, and aggression. The treatment team reached a compromise with the daughter after providing her with education and support. They continued the wheat germ oil and a multivitamin supplement, which appeared safe, even if of limited value. The patient continued taking valproate, 125 mg bid, which reduced her physical aggression and improved resistance to care. All other herbal remedies and zolpidem were discontinued. Balancing traditional therapies with requests for herbal remedies is a common challenge for physicians. The most successful intervention occurs when doctors familiarize themselves with herbal preparations and educate patients and families about the treatments.

A new oscillometric method for pulse wave analysis: comparison with a common tonometric method.

In the European Society of Cardiology-European Society of Hypertension guidelines of the year 2007, the consequences of arterial stiffness and wave reflection on cardiovascular mortality have a major role. But the investigators claimed the poor availability of devices/methods providing easy and widely suitable measuring of arterial wall stiffness or their surrogates like augmentation index (AIx) or aortic systolic blood pressure (aSBP). The aim of this study was the validation of a novel method determining AIx and aSBP based on an oscillometric method using a common cuff (ARCSolver) against a validated tonometric system (SphygmoCor). aSBP and AIx measured with the SphygmoCor and ARCSolver method were compared for 302 subjects. The mean age was 56 years with an s.d. of 20 years. At least two iterations were performed in each session. This resulted in 749 measurements. For aSBP the mean difference was -0.1 mm Hg with an s.d. of 3.1 mm Hg. The mean difference for AIx was 1.2% with an s.d. of 7.9%. There was no significant difference in reproducibility of AIx for both methods. The variation estimate of inter- and intraobserver measurements was 6.3% for ARCSolver and 7.5% for SphygmoCor. The ARCSolver method is a novel method determining AIx and aSBP based on an oscillometric system with a cuff. The results agree with common accepted tonometric measurements. Its application is easy and for widespread use.

Evaluation of cellular fibronectin plasma levels as a useful staging tool in different stages of transitional cell carcinoma of the bladder and renal cell carcinoma.

Reliable markers for both renal cell carcinoma (RCC) and transitional cell carcinoma of the bladder (TCC) are lacking.During tumor progression and invasion components of extracellular matrix (ECM) are degraded and parts of these different components are detectable in plasma. Cellular fibronectin (cFN) represents a well characterized ECM protein. In contrast to fibronectin in plasma produced by hepatocytes (FN) cFN has a total extra domain sequence and occurs in much smaller amounts in the circulation. The aim of our study was to evaluate cFN as a marker and to determine its possible role in clinical staging of TCC and RCC.Blood samples were collected from 30 patients before they underwent transurethral resection of the bladder because of newly diagnosed TCC. Additionally samples were collected from 69 patients with RCC before therapy. Sixty patients with non-malignant urological disorders were recruited as control group. Determination of cFN in plasma was performed by using a highly sensitive time-resolved fluorescence immunoassay (TRFIA).The control group had median cFN plasma levels of 437 ng/ml. Patients suffering from TCC or RCC showed significantly higher cFN levels. In patients with muscle invasive TCC significant higher cFN levels (p < 0.05) could be demonstrated compared to non-muscle invasive TCC. Similar results were found in RCC with significant elevated cFN levels in metastatic RCC (p < 0.005) compared to localized stage of disease. No differences were found concerning tumor grading in both malignancies.In the face of significant elevated cFN levels in TCC and RCC our data underline the important role of cFN. For future investigations the elevated cFN levels in locally progressed and metastastic disease, indicating a clinically useful tool for preoperative staging and postoperative monitoring, are of high interest.

Two sensitive time-resolved fluoroimmunoassays for cellular fibronectin.

Two sensitive sandwich-type immunoassays for determination of cellular fibronectin (cFN) in cell culture supernatants and in human plasma were developed. Both assays used a monoclonal antibody with specificity against the EDA sequence, which is characteristic for the cellular form of human and rat FN. Assay 1 involves binding of FN on gelatin-coated microwells followed by reaction with the anti-cFN antibody, whereas in assay 2 the anti-cFN antibody was immobilized first and detection was with an anti-FN antiserum. Time-resolved fluorescence spectrophotometry with measurement of an Eu3+ chelator after dissociation of the solid-phase complexes with urea/sodium dodecyl sulfate in the presence of excess Eu3+ was the detection method. The detection limit of the new assays was between 2.6 and 4.0 micrograms/L cFN. In serial dilution of human plasma samples, parallelism with the calibration curve was obtained over the whole measuring range (12-1000 micrograms/L) with assay 2, whereas assay 1 had deviations of the dilution curves at concentrations > or = 100 micrograms/L. The between-run CVs for assays 1 and 2 were 11.4% and 7.2%, respectively, at a concentration of 200 micrograms/L (median value of 18 experiments). Respective within-series CVs of 4.3% and 4.7% were obtained at the same concentration. The recovery of added cFN from human plasma was between 90% and 96%.

Logistic-regression model for assessing portal hypertension by measuring hyaluronic acid (hyaluronan) and laminin in serum.

We earlier observed a positive correlation between portal venous pressure (PVP) and the concentration of laminin in serum. Here we investigated whether the diagnostic efficacy could be improved by considering additional analytes and application of multivariate statistical analysis. In 102 patients with fibrotic liver disease of various etiologies we measured PVP as the gradient of the wedged and free hepatic venous pressures and determined the concentrations of hyaluronic acid and laminin in serum. Regression coefficients established by logistic regression in this group were subsequently used to predict portal hypertension (PVP greater than mmHg) in an independent group of 45 patients. By comparison with the known PVP, we obtained a sensitivity of 0.83 (confidence interval: 0.63-0.93) and a specificity of 0.82 (0.61-0.93) for diagnosis of portal hypertension by means of the concentrations of hyaluronic acid and laminin in serum. Application of the model is suggested as a tool for pre-screening and monitoring patients to be subjected to assessment of portal hypertension.

Time-resolved immunofluorometric assays with measurement of a europium chelate in solution: application for sensitive determination of fibronectin.

A novel detection principle applicable for sensitive measurement of molecules of biological interest by time-resolved fluorescence spectrophotometry is described. Our method is based on the quantification of the Eu3+ chelator 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA) in solution in presence of an excess of Eu3+ ions. BCPDA-labeled solid phase complexes obtained by conventional immunoassay procedures are transferred into solution using urea/SDS/Eu3+ as dissociating and fluorescent lanthanide ion reagent. Two 'sandwich-type' assay variants based on the above methodology were realized for the determination of small amounts of fibronectin (FN) in biological fluids. FN is captured from solution by solid phase coated gelatin or a monoclonal antibody, respectively. Rabbit anti-FN antiserum used as second antibody is detected with a biotinylated anti-rabbit IgG antibody. Fluoresence is measured after incubation with streptavidin-BCPDA and dissociation of solid phase complexes as described. Both assays have a detection limit (blank + 3 x SD) of less than 0.5 ng/ml FN, a dynamic range of up to 300 ng/ml, and intraserial coefficients of variation of 4.4 and 6.3%, respectively. Median FN concentrations in saliva of healthy individuals were 104 (gelatin) and 36 ng/ml (double antibody), respectively.

Estimation of the production rates of serum aminoterminal propeptide of type III procollagen and laminin in human fibrotic liver.

The concentrations of laminin, a high molecular weight non-collagenous glycoprotein of basement membranes, and of the N-terminal propeptide of type III procollagen were determined in the serum of the liver outflow vascular region (hepatic vein) and of a peripheral vein (cubital vein) in patients with chronic liver diseases (fibrosis, cirrhosis, unspecified histology; n = 173), in order to determine their secretion rates from the injured livers. The mean levels of laminin (1.84 kU/l) and of procollagen peptide (28.0 micrograms/l) in hepatic vein were significantly higher (about 9.5% at p less than 0.02, and 37% at p less than 0.001, respectively) than those in the periphery (1.68 kU/l and 20.4 micrograms/l, respectively). In chronic liver diseases, however, laminin and procollagen peptide concentrations in the hepatic vein were lower than or equal to those in the cubital vein in 18% and 27% of patients, respectively. The highest regional differences of the concentrations were noted in cirrhotic subjects. The serum levels of laminin (rs 0.93) and of procollagen peptide (rs 0.73) in hepatic and in cubital vein are highly positively correlated (p less than 0.001), but the levels of procollagen peptide in hepatic vein are only weakly but still significantly statistically related with those of laminin (rs 0.446, p less than 0.001). Similarly, the hepatic-cubital venous concentration differences of both proteins are weakly (rs 0.312) but significantly (p less than 0.001) correlated. On the basis of several assumptions we estimated secretion rates from the livers of 120 U.min-1 for laminin, and 5.7 micrograms.min-1 for procollagen peptide.(ABSTRACT TRUNCATED AT 250 WORDS)

Laminin and aminoterminal propeptide of type III procollagen in seminal plasma from fertile and vasectomized men.

The components of the extracellular matrix, laminin and aminoterminal propeptide of type III procollagen (PIIINP) were determined in seminal plasma of 50 patients with vasectomy and of 50 age-matched fertile patients. The concentration of laminin was highly significantly (P < 0.001) elevated in the fertile group as compared to the vasectomy group, whereas the concentrations of PIIINP were not significantly different between these two groups. Only weak correlations were observed between the concentrations of laminin and PIIINP. It is suggested that part of the laminin found in seminal plasma is derived from the ductus deferens, while the source of PIIINP is probably located at an upper part of the urogenital tract.

PIP: The aim was to examine the effect of the resection of the ductus deferens on the pattern of extracellular matrix components in seminal plasma. The components laminin and aminoterminal propeptide of type III procollagen (PIIINP) were determined in seminal plasma of 50 patients with vasectomy and of 50 age-matched fertile patients. Fifty ejaculates from normal fertile men and 50 ejaculates from patients with vasectomy were randomly selected from a pool of clinical specimens at the Kantonsspital Aarau, Switzerland. Age ranges were between 19 and 43 years. Laminin was measured with a competitive radio immunoassay using a rabbit antiserum against the pepsin-resistant fragment P1 of human laminin. PIIINP was measured with a new radioimmunoassay using monoclonal antibodies against the aminoterminal propeptide in a 2-stage sandwich assay. Laminin and PIIINP could be detected in all seminal plasma of the fertile group and the vasectomy group, respectively. The median concentration of laminin in seminal plasma was 1.98 kU per liter in the fertile group and 1.25 kU per liter in the vasectomy group. The median concentration of PIIINP in the seminal plasma of the fertile group was 0.59 kU per liter and 0/52 kU per liter in the vasectomy group. For laminin the concentration levels in the fertile and the vasectomy group were significantly different (P or= 10 -7). No significant differences were obtained for PIIINP (P 0.05). The concentration of laminin was very significantly (P 0.001) elevated in the fertile group as compared to the vasectomy group, whereas the concentration of PIIINP were not significantly different. In the combined groups, a weak correlation between laminin an PIIINP was observed (P 0.05). This correlation was even lower in the fertile group (P 0.05), whereas within the vasectomy group it was slightly higher. If results can be verified in larger groups of patients with aspermia caused by testicle disorders and by a block of the ductus deferens, the determination of laminin in seminal plasma might prove useful as an adjuvant diagnostic measure.

The concentration pattern of laminin, hyaluronan, and aminoterminal propeptide of type III procollagen in seminal fluid.

The extracellular matrix components laminin, N-terminal propeptide of type III procollagen (PIIINP) and hyaluronan (HA) were determined in seminal fluids of 119 patients submitted for diagnosis of infertility. The concentrations of laminin and HA, but not those of PIIINP, were elevated in seminal fluid in comparison to their ranges of concentration in normal sera. Only weak correlations were observed between the concentrations of the three matrix components. The concentration of HA was negatively correlated with sperm count and ejaculate volume. Laminin was positively correlated with sperm count, the age of patients, and highly significantly with the concentrations of acrosin. A highly significant positive correlation was also found between PIIINP and fructose. By analysis of variance it could be shown that patients with azoospermia and oligozoospermia have significantly higher levels of HA than those with normospermia. Patients with terato- and asthenozoospermia showed no characteristic pattern of the matrix components.

Influence of reagent composition on atypical pseudocholinesterase activity measurement: comparison of a manual and an automated method and implications for routine.

In determining activities of atypical pseudocholinesterase using a Hitachi 737 and a Cobas Bio analyser, respectively, we repeatedly found discrepancies between the enzyme activities measured by both instruments which were presumably due to differences in the reagent composition (Hitachi 737: Boehringer Mannheim Automated Analysis for BM/Hitachi Systems; Cobas Bio: Boehringer Mannheim Monotest). We consider the present results as a strong hint for manufacturers towards generally declaring all components of the chemical systems they produce and as a warning for users when interchanging apparently identically composed sets of reagents.

Efficacy of serum laminin measurement for diagnosis of fibrotic liver diseases.

We examined the efficacy of laminin assay in serum for diagnosis of fibrotic liver diseases. Values for subjects with liver disease significantly (P less than 0.05) exceeded those for healthy subjects and patients with nonhepatic diseases. At a cutoff value of 1.45 kilo-units(arb.)/L (approximately 330 micrograms/L) and an assumed prevalence of fibrotic liver diseases of 0.5, positive and negative predictive values of the test were 0.97 and 0.83, respectively, for the comparison with a healthy reference population and 0.81 and 0.80 for nonhepatic diseased patients. Increases in laminin concentration were positively correlated with the extent of fibrotic transition of the liver. Discrimination between fibrotic and cirrhotic stages of chronic liver diseases by means of laminin assay was better than with the amino-terminal propeptide of type III procollagen. According to the criteria of diagnostic efficacy, we conclude that determination of laminin in serum improves the possibilities of clinical-chemical diagnosis of liver fibrosis and cirrhosis. However, as commonly true for other biochemical tests, determination of laminin cannot replace conventional diagnostic methods.

[The clinical value of laminin determination in advanced liver cirrhosis]. / Klinischer Wert der Lamininbestimmung bei fortgeschrittener Leberzirrhose.

OBJECTIVE: To test prospectively whether serum laminin levels, which is taken to indicate portal hypertension, can predict the occurrence of severe complications in advanced cirrhosis of the liver. PATIENTS AND METHODS: In 38 patients (21 men, 17 women; mean age 55.6 +/- 13.4 years) with liver fibrosis (n = 4) or liver cirrhosis (n = 34) serum laminin was measured by a commercially available radioimmunoassay (Behring, Marburg). The severity of liver cirrhosis was graded according to the Child-Pugh-Christensen criteria. Portal hypertension was assessed by standard endoscopic methods and portal-vein duplex sonography. Within a mean observation period of 12.5 +/- 3.5 months, the following were used as signs of severe clinical complications of liver cirrhosis: stages III and IV of hepatic coma, treatment-refractory ascites, portal vein thrombosis and death due to multi-organ failure. Acute bleeding from oesophageal varices was confirmed by emergency endoscopy. RESULTS: At laminin concentrations of 3.25 +/- 0.20 U/ml there was a highly significant correlation (P < 0.001) with complications of liver cirrhosis. Using 2.6 U/ml as the critical level, the occurrence of severe complications had a positive predictive value of 0.80 with a sensitivity and specificity of 0.71 and 0.86 respectively. This means that a patient who, at the beginning of the study period, had a raised laminin concentration, had a relative risk of 2.65 (1.41-4.97) for later severe complications. CONCLUSION: Serum laminin concentration has a diagnostic efficiency of 0.79 as a prognostic indicator and can thus serve as a valuable addition to the Child-Pugh-Christensen classification of liver cirrhosis.

Serum laminin and hyaluronan in liver cirrhosis: markers of progression with high prognostic value.

BACKGROUND/AIMS: In a prospective study with a mean follow-up period of 12.5 +/- 3.5 months, we investigated the extracellular matrix components laminin and hyaluronan in serum for their diagnostic value in portal hypertension and in clinically severe complications of progressive liver cirrhosis. METHODS: In 38 patients with liver fibrosis (n = 4) and cirrhosis (Child A: n = 17, B: n = 7, C: n = 10), the serum concentrations of laminin and hyaluronan were determined. Portal hypertension was assessed by endoscopic control of the esophageal varices and by Doppler sonography of the portal blood flow. RESULTS: Neither laminin nor hyaluronan correlated with portal hypertension, but highly significantly increased (p < 0.001) concentrations of 3.25 +/- 0.2 U/ml (laminin) and 493 +/- 248 ng/ml (hyaluronan) were found in patients with complications of liver cirrhosis when compared to those without complications (Ln: 2.13 +/- 0.26 U/ml, HA: 206 +/- 184 ng/ml). At cut-off levels of 2.6 U/ml (laminin) and 200 ng/ ml (hyaluronan), the diagnostic sensitivity and specificity for severe complications of liver cirrhosis was 0.71 and 0.86 (Ln) and 0.90 and 0.67 (HA), respectively. The positive predictive values were of 0.8 (laminin) and 0.6 (hyaluronan). The relative risk of patients presenting elevated concentrations of laminin or hyaluronan at the start of the study for later development of severe complications was 2.7. CONCLUSIONS: Both parameters, especially serum laminin, can be used as prognostic markers in addition to the Child criteria in liver cirrhosis.

On the performance and reliability of mechanized urine teststrip measurement in comparison with visual reading.

Prescreening of urine specimens by teststrips is a valuable procedure for reducing the work load of the urine analysis laboratory: positive results for leukocytes, erythrocytes (haemoglobin), protein, and/or nitrite are widely used to select pathological specimens for subsequent microscopic examination. By standardization of the measurement conditions, mechanized teststrip reading is claimed to give more reproducible results than conventional techniques. To assess their ability to improve urine prescreening, especially with regard to the comparability of the results, the practical and analytical performance of three commercially available analysers (Rapimat II/T from Behringwerke AG, Urotron RL9 from Boehringer Mannheim GmbH, and Clinitek 200 from Ames/Bayer Diagnostic) was compared with visual reading. Analytical criteria were assessed using routine urine samples, while reproducibility was tested by repeated analysis of three different commercial control urines (Kova Trol from Madaus). A mean imprecision between 3% and 11.9% was found for the mechanized dipstick reading which was comparable to that found with visual examination (4.5% with Combur9-teststrips, Boehringer Mannheim GmbH). Due to the crude classification of the results, the different analysers as well as the visual technique gave quite different distributions for each of the semiquantitative parameters in the same urine samples. Even if statistical analysis was restricted to the frequency of positive results only, significant differences (chi 2-test, p less than 0.001-0.05) between methods were obtained, but these differences could not be attributed to one method alone. Using microscopic sediment analysis as reference, pathological urines were detected with a comparable sensitivity/specificity: Urotron 0.85/0.84, Rapimat 0.91/0.67, Clinitek 0.82/0.81, and duplicate visual reading 0.88/0.67 and 0.91/0.93. Mechanized teststrip reading had no obvious advantage with respect to the time required. We conclude: (i) no improvement in analytical performance or in speed of analysis could be claimed for mechanized methods in comparison with visual reading; (ii) mechanized teststrip reading might decrease the work load of the urine laboratory if integrated into a computerized laboratory system; (iii) mechanized teststrip reading will become analytically advantageous over visual reading if a more refined classification of the results is achieved.

Follow-up of the serum levels of extracellular matrix components in acute and chronic pancreatitis.

Time-dependent serum concentrations of extracellular matrix proteins were studied in 32 patients with pancreatitis in order to find potential markers of the reparative response during the disease. Patients were subdivided by clinical and biochemical criteria: severe acute pancreatitis (n = 10), moderate acute pancreatitis (n = 17), and acute attack of chronic pancreatitis (n = 5). Serum and plasma samples were collected on days 1-7, 10, 14, and 21 for measurements of the aminoterminal propeptide of type III procollagen (PIIINP), hyaluronic acid, laminin, fibronectin, and routine clinical-chemical parameters. During an acute attack of chronic pancreatitis all parameters were within the reference range. In moderate acute pancreatitis concentrations of PIIINP, laminin, and hyaluronic acid fluctuated around the upper reference limit, but declined to mid-normal levels at day 21. In severe acute pancreatitis all three parameters increased. In patients who died as a consequence of sepsis and multi-organ failure the increase in PIIINP, laminin and hyaluronic acid was much more pronounced and paralleled by a decrease in plasma concentrations of fibronectin. In conclusion, this study revealed a relation between the severity of acute pancreatitis and the increase in serum concentrations of extracellular matrix components, especially PIIINP.