W. Prellwitz

Biliary adenocarcinoma: Characterisation of three new human tumor cell lines

Three human cell lines from adenocarcinomas of the extrahepatic biliary tract were established in permanent tissue culture. Mz-ChA-1 and Mz-ChA-2 were cultured from mechanically dissociated gallbladder adenocarcinoma metastases and SK-ChA-1 was grown from malignant ascites of a patient with primary adenocarcinoma of the extrahepatic biliary tree. Cell doubling times in tissue culture are 3-4 days for Mz-ChA-1 and approximately 2 days for Mz-ChA-2 and SK-ChA-1. All three tumour cell lines were successfully transplanted to nude mice, inducing progressive tumour growth. Histologically, nude mouse tumours resembled the original adenocarcinomas. In vitro formation of gland-like structures were regularly seen in Mz-ChA-1 and Mz-ChA-2 but only occasionally in SK-ChA-1. All three cell lines formed contacts through interdigitating processes with desmosomes and junctional complexes. On scanning electron microscopy, an abundance of microvilli was seen at the cell surfaces. Chromosome analyses of all three tumour cell lines showed a wide range of numerical abnormalities and presence of marker chromosomes. Mz-ChA-1 appears to be highly differentiated with cells producing mucus. Mz-ChA-2 synthesizes components of complement C2, C3 and C5, while Mz-ChA-1 and SK-ChA-1 produce only C3 in detectable quantities. In addition, Mz-ChA-2 supernatants are positive for ferritin and alpha 1-fetoprotein, but not CEA; while Mz-ChA-1 and SK-ChA-1 produce only CEA. Supernatants of all three cell lines are positive for N-acetyl neuraminic acid (NANA), phosphohexoisomerase (PHI) and LDH, and negative for alpha 2-macroglobulin, alpha 1-anti-trypsin, gamma-GT, AP, coeruloplasmin, haptoglobin and albumin. A high cloning efficiency renders these new tumour cell lines suitable for continued studies on clonal heterogeneity in malignant tumours. The establishment of these cell lines in tissue culture facilitates further studies on the biology of upper gastrointestinal tract cancer in man.

Bestimmung der Aktivität von Creatinkinase MB im Serum unter Verwendung inhibierender Antikörper

SummaryA new method for the determination of creatine kinase-MB activity in the serum is presented. The principle of this method is the direct measurement of the activity of creatine kinase M subunits by inhibiting antibodies. The total test procedure takes 15 min. In the sera of all the 83 patients tested, who have clinically proven myocard infarction, creatine kinase-MB activity can be measured between the 6th and 28th hour after infarction. At the time of maximum total creatine kinase activity the percentage of creatine kinase-MB activity is between 6 and 17%, the mean value being 8%. In cases of emergency this method can be used for the differential diagnosis of elevated total creatine kinase activities of unknown origin.ZusammenfassungEs wird über eine neue Methode zur quantitativen Bestimmung der Creatinkinase MB-Aktivität im Serum berichtet. Die Methode beruht auf einer direkten Messung der Aktivität der Creatin-kinase-Untereinheit B nach Hemmung der Aktivität der Creatinkinase-Untereinheit M durch inhibierende Antikörper und benötigt zur Durchführung 15 min. Bei allen 83 untersuchten Patienten mit klinisch gesichertem Myokardinfarkt konnten zwischen der 6. und 28. Stunde nach Infarkteintritt Creatinkinase MB-Aktivität gemessen werden. Der Creatinkinase MB-Anteil zum Zeitpunkt der höchsten Creatinkinase-Gesamtaktivität betrug 6–17%, im Mittel 8%. Diese Methode ermöglicht daher in der Notfalldiagnostik eine Differentialdiagnose unklarer Creatinkinase-Gesamtaktivitäts-Erhöhungen.A new method for the determination of creatine kinase-MB activity in the serum is presented. The principle of this method is the direct measurement of the activity of creatine kinase M subunits by inhibiting antibodies. The total test procedure takes 15 min. In the sera of all the 83 patients tested, who have clinically proven myocard infarction, creatine kinase-MB activity can be measured between the 6th and 28th hour after infarction. At the time of maximum total creatine kinase activity the percentage of creatine kinase-MB activity is between 6 and 17%, the mean value being 8%. In cases of emergency this method can be used for the differential diagnosis of elevated total creatine kinase activities of unknown origin.

Effect of long‐term treatment with GH on bone metabolism, bone mineral density and bone elasticity in GH‐deficient adults

Adults with GH deficiency (GHD) commonly have subnormal bone mineral density (BMD), and have been reported to have an increased risk of fractures. It has been suggested that GH replacement therapy may have beneficial effects on bone in such patients. The aim of this study was to investigate the effects of long‐term GH replacement therapy on bone metabolism, BMD and bone elasticity in adults with GHD.

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.

The Organ Distribution of Selenium in German Adults

The selenium concentrations were determined in liver, kidney, skeletal muscle, heart, brain, prostate, testis, bile, lung, and spleen of German traffic accident victims. In addition the nitrogen and phosphorus contents were determined in the same organs and tissues. On a per-weight unit basis, the highest selenium concentration was found in kidney. However, this corresponds to only 4% of the total body selenium. Most of the whole body selenium (50%) is present in skeletal muscle, which thus appears to act as a selenium storage organ. However, there is also evidence that selenium is required for muscle function. In plasma and interstitial fluid, .450 mg of Se, or 7.5% of the total body selenium is present. A comparison of the organ Se concentrations of the German traffic accident victims with the selenium concentrations of the same human organs as reported in different countries indicates that the organ concentrations of West Germans are comparable to that of the population of New Zealand, a low-Se country, and significantly lower than that observed in the organs of American, Canadian, and especially Japanese subjects. The international comparison of the organ selenium concentrations also revealed that the selenium uptake of kidney is higher at low- and adequate dietary Se intakes and lower if the dietary Se supply is high, as is the case for Japanese subjects. Estimates of the daily excretion of selenium with the bile indicate that the amounts are three times higher than the daily urinary losses and in the same order of magnitude as the daily dietary selenium intakes. Enterohepatic reabsorption of selenium from the bile appears to be a significant mechanism of conserving dietary selenium and to maintain Se balance at comparatively low dietary Se intakes.

Retinal vascular occlusion and deficiencies in the protein C pathway

PURPOSE To report abnormalities in the protein C pathway and other vascular occlusion risk factors in patients with retinal vascular occlusion. METHODS In a study, we investigated 76 consecutive patients who had in-patient evaluation of venous or arterial retinal vascular occlusion. All patients underwent comprehensive tests for coagulation disorders including determinations of protein C, protein S, lupus anticoagulants, and resistance to activated protein C and were screened for vascular disease risk factors. Resistance to activated protein C was confirmed by a polymerase chain reaction method to detect the specific factor V R506Q mutation. For comparative purposes, we also screened 209 consecutive inpatients with deep vein thrombosis from the same geographic region for resistance to activated protein C as well as protein C and protein S deficiencies. RESULTS Ten (29%) of 35 patients with central retinal vein occlusion (CRVO) had factor V R506Q mutation. The factor V R506Q mutation was detected in four (19%) of 21 patients with branch retinal vein occlusion. The higher frequency in factor V R506Q mutation compared with the expected 9% mutation prevalence in a white population was highly significant for the central retinal vein occlusion group but not for the branch retinal vein occlusion group. In all patients with resistance to activated protein C, the factor V R506Q mutation was detected; 16 were heterozygous, one homozygous. No cases of lupus anticoagulants, protein C, or protein S deficiencies were detected. Forty (19%) of 209 patients with deep vein thrombosis were carriers of the factor V R506Q mutation. CONCLUSIONS The prevalence of the factor V R506Q mutation is similar in patients with central retinal vein occlusion and patients with deep vein thrombosis and represents a relevant risk factor. Screening for this mutation is therefore recommended in all patients with central retinal vein occlusion.

Influence of HMG–CoA reductase inhibitors on markers of coagulation, systemic inflammation and soluble cell adhesion

BACKGROUND Beneath its lipid-lowering properties additional non-lipid effects of statin therapy are discussed. We therefore examined the impact of statins on laboratory markers of coagulation, inflammation and soluble cell adhesion to further explore these effects in 950 hospitalised patients with angiographically proven CAD. METHODS AND RESULTS Although no significant differences were found in total cholesterol, LDL and HDL and triglyceride levels a statistically lower value in 277 statin-treated patients was found for von Willebrand factor [162(130/224) vs. 208(154/283)%, P=0.0001], leukocyte count [6.9(5.8/8.4) vs. 7.3(6.1/9.4)/nl, P=0.0005], high sensitive CRP [4.3(1.8/10.8) vs. 7.6(2.8/20.0) mg/dl, P=0.0001], interleukin-6 [9.5(5.1/18.7) vs. 14.4(7.2/28.1) mg/dl, P=0.0001] and soluble p-selectin [112.6(82.0/146.0) vs. 127.8(93.8/162.4) mg/dl, P=0.001] compared to 673 patients without statin therapy. This result was confirmed in a subgroup of 510 patients matched for age, gender and percentage of acute coronary syndromes. CONCLUSIONS In statin treated patients significantly lower levels of coagulation, systemic inflammation and soluble cell adhesion markers were found. Therefore the effect of statin therapy may also be mediated by additional non-lipid-lowering effects.

Congestive cardiomyopathy and the selenium content of serum

A deficiency of selenium is suspected to be involved in the pathogenesis of congestive cardiomyopathy. Therefore the serum selenium content of 20 patients with proven congestive cardiomyopathy was measured and compared to that of a healthy control group. The serum selenium content of the patients with cardiomyopathy was found to be different from that of the healthy control group. The mean value of selenium in serum for the control group was 80.1 micrograms Se/1 (SD +/- 13.2) within a range of 53 and 117 micrograms Se/1. From the 20 patients with congestive cardiomyopathy six patients showed selenium concentrations in the normal value range of the control group; in the serum of 14 patients a distinct lower selenium content was found (mean value 47.8 micrograms Se/1 (SD +/- 16.2)) within a range of 23 and 70 micrograms Se/1. A positive correlation was found between serum selenium content and the left ventricular ejection fraction. Our results suggest that a deficiency of selenium may be present in a number of patients with congestive cardiomyopathy.

Intracoronary thrombolysis with an acylated streptokinase-plasminogen activator (BRL 26921) in patients with acute myocardial infarction

The fibrinolytic efficacy and systemic effects on coagulation variables of intracoronary administration of an acylated streptokinase-plasminogen complex (BRL 26921) were assessed in 23 patients with an acute transmural myocardial infarction. The infarct vessel was totally occluded in 22 patients and subtotally stenosed in 1 patient. Reperfusion was achieved in a total of 17 patients (74%), in 2 patients with the use of a guide wire. Reperfusion time in those patients treated with BRL 26921 alone amounted to 42 +/- 37 minutes. Reocclusion occurred in two patients subsequently. Four patients died; in two of these, intracoronary thrombolysis was unsuccessful. Reptilase time increased from 13 +/- 3 to 49 +/- 31 seconds (p less than 0.001), fibrinogen levels decreased from 280 +/- 65 to 126 +/- 76 mg% (p less than 0.001). Factor V decreased from 96 +/- 11 to 53 +/- 26% (p less than 0.001), and factor VIII from 99 +/- 1 to 55 +/- 36% (p less than 0.001). Peripheral hyperplasminemia, defined as a reduction of fibrinogen (less than 100 mg%) with a reduction of factor V and VIII (less than 75%) simultaneously occurred in eight patients. Six (75%) of these 8 patients demonstrated reperfusion, whereas 9 (64%) of 14 patients without peripheral hyperplasminemia were also successfully reperfused. Bleeding complications occurred in two patients who demonstrated hyperplasminemia. Thus, effective intracoronary thrombolysis could be achieved with only minor effects on peripheral coagulation variables in the majority of patients.

The renal excretion of selenium.

The excretion of selenium in urine was determined in West German healthy volunteers. Women excrete 17.7±4.2 μg Se/d and men 19.0±9.0 μg Se/d. The daily selenium excretion per gram creatinine is 13.5±3.8 μg Se/g crea for women and 9.8±3.3 μg Se/g crea for men. The clearance of selenium from the plasma is calculated with 0.18 mL/min. The selenium excretion per day is positively correlated with the 24h excretion of urea and creatinine. The correlation of the selenium excretion with the urea excretion is most probably owing to the fact that the selenium intake of West Germans is linked primarily to foods with high protein contents. That the selenium excretion is directly correlated with the creatinine excretion is an indicator that the muscle, which accounts for nearly 50% of the whole body selenium in West German adults, influences the selenium excretion in urine. The positive correlation of the selenium excretion with the potassium excretion also indicates that the muscle mass contributes significantly to the selenium excretion in urine. Another indicator that the selenium excretion is influenced by the muscle is that after intensive muscular activity (running), selenium excretion is enhanced. The 24h selenium excretion is dependent on the glomerular filtration rate of the kidney characterized by the creatinine clearance. This result is important, because if the selenium excretion is used as parameter for the selenium status of humans, the kidney function should be known. This is a limitation for the use of the urinary selenium excretion as parameter for the selenium status. This is especially important for patients whose glomerular filtration rate is low. The 24h selenium excretion is further influenced by the 24h urine volume. Selenium losses via urine may be concomitant with protein losses in urine.