Cornelia Spamer

Survival benefit of patients with inoperable hepatocellular carcinoma treated by a combination of transarterial chemoembolization and percutaneous ethanol injection: A single-center analysis including 132 patients

Hepatocellular carcinoma (HCC) is one of the most severe sequelae of chronic liver disease. The only potentially curative therapeutic options are surgical resection and orthotopic liver transplantation. In most HCC patients, however, at clinical presentation the tumors are unresectable because of multicentricity or poor hepatic functional reserve due to pre‐existing cirrhosis or not transplantable because of too advanced tumor stage or severe co‐morbidity. In clinical practice, therefore, percutaneous ethanol injection (PEI) and transarterial chemoembolization (TACE) are widely used non‐surgical therapeutic strategies. We prospectively analyzed the clinical factors determining the prognosis of 132 inoperable HCC patients and assessed the feasibility, therapeutic efficacy and safety of PEI, TACE and a combination thereof. Mean age of patients was 64 years; 95% of patients had liver cirrhosis and 39% were Okuda stage I, 48% stage II and 13% stage III. Fifteen patients were treated by PEI (group 1), 33 by TACE (group 2), 39 by TACE and PEI (group 3) and 45 received best supportive care (group 4). Survival correlated with the Child‐Pugh class of liver cirrhosis and the Okuda stage of HCC. Favorable prognostic parameters were alpha‐fetoprotein (AFP) levels <100 ng/ml and absence of portal vein thrombosis. Median survival time was 18 months in group 1 [interquartile range (IQR) 10–19], 8 months in group 2 (IQR 5–15), 25 months in group 3 (IQR 13–36) and 2 months in group 4 (IQR 1–9). Multivariate analysis revealed that patients treated with a combination of TACE and PEI have a significantly better survival than patients receiving either PEI or TACE only (p = 0.001). Patients with inoperable HCCs treated by the combination of TACE and PEI have a clear survival benefit. A favorable outcome can be expected in patients with compensated cirrhosis, a low Okuda stage, a baseline AFP level <100 ng/ml and absence of portal vein thrombosis. Int. J. Cancer (Pred. Oncol.) 79:601–605, 1998.

Identification of the Zn2+ binding region in calreticulin.

Calreticulin binds Zn2+ with the relatively high affinity/low capacity. To determine the location of the Zn2+ binding site in calreticulin different domains of the protein were expressed in E. coli, using the glutathione S‐transferase fusion protein system, and their Zn2+‐dependent interaction with Zn2+‐IDA‐agarose were determined. Three distinct domains were used in this study: the N + P‐domain (the first 290 residues); the N‐domain (residues 1–182) and the proline‐rich P‐domain (residues 180–273). The N + P‐domain bound to the Zn2+‐IDA‐agarose and were eluted with an increasing concentration of imidazole. The N‐domain also bound 65Zn2+ as measured by the overlay method. The P‐domain did not interact with the Zn2+‐IDA‐agarose and it did not bind any detectable amount of Zn2+. Chemical modification of calreticulin with diethyl pyrocarbonate indicated that five out of seven histidines were protected in the presence of Zn2+ but they were modified by diethyl pyrocarbonate in the absence of Zn2+ suggesting that these residues may be involved in Zn2+ binding to calreticulin. We conclude that Zn2+ binding sites in calreticulin are localized to the N‐domain of the protein, region that is not involved in Ca2+ binding to calreticulin.

Immuno-gold electron microscopical detection of heat shock protein 60 (hsp60) in mitochondria of rat hepatocytes and myocardiocytes

We characterize the specificity of a polyclonal antibody against heat shock protein 60 (hsp60) and present an application for ultrastructural localization studies of this protein. The antibody was obtained from an IgG fraction (AB 121) originally raised against the calcium binding protein calsequestrin by immunoabsorption on isolated rat liver hsp60. As shown by partial N-terminal amino acid sequence analysis of immunoprecipitated proteins AB 121 contained reactivities against hsp60, calsequestrin and the glycoprotein fetuin. In rat heart AB 121 recognized calsequestrin and hsp60. In human and rat liver the only reacting protein was hsp60. In rat erythrocytes the antibody bound to 61 kDa and 58 kDa isoforms of fetuin. According to published data no amino acid sequence homologies nor common motifs are found between calsequestrin, hsp60 and fetuin. As the first application the anti-hsp60 antibody was used for immuno-gold electron microscopical localization of hsp60: in myocardiocytes and hepatocytes of the rat strong labelling was obtained exclusively in mitochondria. No extramitochondrial structures were labelled. The specificity of the antibody and its ability to be visualized by immuno-gold electron microscopy offers the possibility to study the expression of this protein in the liver and in other organs. Possible clinical applications of these studies are discussed, since hsp60 could be a target antigen of autoantibodies in diseases such as autoimmune hepatitis, primary sclerosing cholangitis or primary biliary cirrhosis.

The phosphoprotein intermediate of a Ca2+ transport ATPase in rat liver endoplasmic reticulum

Smooth endoplasmic reticulum vesicles from rat liver display an ATP-supported Ca2+ transport which is mediated by a (Ca2+ + Mg2+)-ATPase. During the catalytic cycle the terminal phosphate from ATP is incorporated to form an acid-precipitable reaction product(118 000-Mr in SDS-gel electrophoresis) with stability characteristics of an acylphosphate. Comparative studies with sarcoplasmic reticulum vesicles from fast-twitch skeletal muscle suggest that the 118 000-Mr phosphopeptide may be identified with the phosphorylated reaction intermediate of a Ca2+ transport ATPase in endoplasmic reticulum, similar to that in sarcoplasmic reticulum of muscle.

Clinical significance of abnormalities of the gastrointestinal tract detected by abdominal ultrasound.

In order to define the clinical significance and the need for further clinical work-up in patients where abnormalities of the stomach or bowel are found by ultrasound, we performed a prospective study on 100 patients with such findings. Of all patients, 35% were found to have a malignant tumor, 73% had a diagnosis as made by reference methods which was probably (18%) or definitively (55%) related to the US finding. Eighteen percent had definitive false positive findings, in 9% no final diagnosis was obtained. Thus, a positive predictive value of 80% was calculated for the US finding of a mass or a target sign related to bowel or stomach in US. The positive predictive value was lowest for target signs related to the bowel (75%) and highest for bowel conglomerates (100%). Patients with target signs of the stomach, bowel conglomerates, or masses were more likely to have malignant disorders (72%) than those with target signs related to the bowel (16%). We conclude from this study that the ultrasound finding of a target sign or a mass related to stomach or bowel has a high clinical relevance and should in any case worked up by appropriate investigations when clinical consequences are possible.

Mechanism of the calcium pump in the endoplasmic reticulum of liver: phosphoproteins as reaction intermediates.

Microsomal fractions, highly enriched with endoplasmic reticulum of rat and human liver exhibit Ca2+ uptake catalyzed by a Ca2+-pumping ATPase. The mechanism of Ca2+-translocation involves: (i) reversible Ca2+-dependent formation of an acyl-phosphoenzyme intermediate (Mr 116,000 to 118,000) with bound Ca2+, which in the reversed reaction can transphosphorylate its Pi to ADP to re-synthesize ATP; (ii) reversible transition of the ADP-reactive phosphoenzyme into an isomer without bound Ca2+, not further reactive to ADP; (iii) hydrolytic cleavage, stimulated by Mg2+, K+, and ATP of the ADP-unreactive phosphoenzyme with liberation of Pi. By analogy to a mechanism proposed for the Ca2+ pump of sarcoplasmic reticulum, the translocation of Ca2+ to and dissociation from the inner side of the membrane is suggested to occur by a conformational change, coupled with a decrease in Ca2+-affinity of the phosphoenzyme during its transition into the ADP-unreactive isomer. With CaATP as the effective substrate the reactions proceed normally but at a considerably slower rate.

Diagnostic value of ultrasound in duodenal stenosis.

Radiology and endoscopy are usually used to diagnose stenosis of the duodenum, but the cause of the stenosis may still escape detection. Three pathologic processes — multiple duodenal polyps, duodenal wall cysts, and annular pancreas — are presented to demonstrate how ultrasound, under specific conditions, can provide decisive diagnostic information. The cause of the stenosis may be made accessible to sonographic study through administration of oral fluids and intravenous hyoscin-n-butylbromid (Buscopan®).