[Three-dimensional reconstruction of solid tumors : Morphological evidence for tumor heterogeneity]. / Dreidimensionale Rekonstruktion solider Tumoren : Morphologische Evidenz für Tumorheterogenität.

BACKGROUND: In histopathological routine diagnostics, three-dimensional tissue samples are analyzed histologically and/or immunohistochemically in two-dimensional sectional planes due to the high expenditure of time and the lack of digitization possibilities. AIM: Here, we demonstrate the application of three-dimensional reconstruction to solid tumors and analyze inter-/intratumoral heterogeneity with respect to epithelial-mesenchymal transition (EMT). METHODS: Tissue samples from pancreatic, lung, colorectal, and breast cancers as well as colorectal liver metastases were serially processed in 4µm sections. For individual analyses, alternating stains (cytokeratin AE1/3, zinc finger E­box-binding homeobox 1 (ZEB1), eCadherin) were performed. Subsequently, the tumor cells were analyzed for their morphology (epitheloid amoeboid, mesenchymal) and the expression of ZEB1 and eCadherin. For statistical analysis, all tumor cell aggregates were hierarchically annotated and analyzed. RESULTS: Tumor buds are predominantly associated with the main tumor mass. Furthermore, a shutteling of eCadherin could be observed within tumor cell aggregates smaller than nine cells. ZEB1 is only increasingly expressed in tumor cell groups smaller than five cells. CONCLUSIONS: The initial tumor budding and the subsequent decoupling of the tumor bud from the main tumor mass is most likely a two-part process. However, the EMT is not statistically significantly increased within the tumor bud detached from the main tumor mass. It could be shown that the currently valid and known definition of a tumor bud as a cell cluster of less than or equal to five cells cannot be completely classified in the concept of EMT represented by eCadherin and ZEB1.

Comparison of immunophenotypes of primary breast carcinomas and multiple corresponding distant metastases: an autopsy study of 25 patients.

Phenotypical change in metastatic breast carcinoma has widely been accepted as an inherent biological feature rather than technical fault. We analyzed the immunohistochemical phenotype and histopathological features of 25 primary breast carcinomas and 90 corresponding distant metastases in 23 organs retrospectively. Histological slides were reviewed for prognostic and predictive factors. Overall, metastases were more similar to each other and often differed from the primary tumor. We created a 3-step grouping system based on the localization of metastases. Regions: tumors metastasizing to the abdominal region were likely to lose ER (p = 0.002); we detected loss of PR in metastases to the thorax (p = 0.039) and abdomen (p < 0.001). Organ systems: loss of ER and PR was observed in metastases to the gastrointestinal system (p = 0.026 and p = 0.001, respectively), in the respiratory system only the loss of PR was significant (p = 0.05). Individual organs: the primaries were likely to lose the hormone receptors in liver metastases (ER p = 0.026; PR p = 0.004). In lung metastases only loss of PR was apparent (p = 0.049). We did not observe significant change in HER2 status, regarding Ki67 change occurred only in bone metastases compared to the primary (p = 0.048). 7/25 patients' distant metastases had heterogeneous immunoprofiles. The later the metastasis was discovered the more likely it had a differing IHC profile compared to the primary tumor, patients who had longer OS had a higher chance to develop a discordant metastasis. Immunoprofile of metastases may differ from primary breast cancer and from each other, probably resulting in different response to therapy.

Chemosaturation with percutaneous hepatic perfusions of melphalan for hepatic metastases: experience from two European centers.

PURPOSE: Chemosaturation with percutaneous hepatic perfusion (PHP; Hepatic CHEMOSAT(®) Delivery System; Delcath Systems Inc, USA) is a minimally invasive, repeatable regional therapy for unresectable hepatic metastases. It uses a system of catheters and filters to isolate hepatic venous blood from the systemic circulation, allowing delivery of high-dose chemotherapy to the hepatic artery. Effluent hepatic venous blood is filtered before being returned to the systemic circulation, thereby reducing exposure to chemotherapy. We describe our experiences with chemosaturation-PHP at 2 European centers. MATERIALS AND METHODS: 14 patients presented unresectable hepatic metastases from solid tumors; 13 received 1 - 3 sessions of chemosaturation-PHP. Melphalan 2.0 (n = 1) or 3.0 (n = 12) mg/kg was given as a 30-minute infusion into the hepatic artery. 12 patients were evaluable for tumor response. RESULTS: One complete (cholangiocarcinoma, n = 1) and 6 partial responses (ocular, n = 3 or cutaneous melanoma, n = 3) were observed, 5 patients had stable disease (ocular melanoma, n = 3; breast cancer, n = 1; gastric cancer, n = 1). Mild to moderate filter-related toxicity (i. e. thrombocytopenia, anemia) was observed immediately post-procedure. Grade 3/4 melphalan-related pancytopenia developed after 1 - 2 weeks. All hematological events were managed effectively with transfusions and/or other supportive measures. The new high-efficiency filter showed milder toxicity and faster recovery. In one case, chemosaturation-PHP was abandoned prematurely due to heparin-induced vaginal bleeding, and one patient died due to retroperitoneal hemorrhage from heparin anti-coagulation. CONCLUSION: Chemosaturation-PHP for non-resectable liver metastases is a feasible treatment option when performed by an experienced multi-disciplinary team. It may be a promising regional therapy for patients with no effective treatment options.

Xenon anesthesia for liver transplant surgery: a report of four cases.

It is well established that patients presenting for orthotopic liver transplantation pose challenging surgical and anesthesiological problems. Intraoperatively, severe hemodynamic instability due to profuse bleeding and acute cardiomyopathy during reperfusion are major concerns. In addition, ischemia-reperfusion injury can compromise postoperative graft function. Xenon, with its potential to maintain hemodynamic stability, preserve cardiac function, and protect the liver graft of the recipient, seems to be a promising anesthetic agent for liver transplant surgery. To date, xenon has not been used as an anesthetic in liver transplantations. We therefore have reported our initial experience with four patients who underwent orthotopic deceased donor liver transplantation under xenon anesthesia. Although all patients had advanced liver disease and experienced significant intraoperative bleeding, their intraoperative courses, including reperfusion, under xenon anesthesia were remarkably stable. The patients required only moderate, temporary catecholamine support, which was withdrawn at the end of the surgery. Xenon anesthesia for liver transplant procedures proved to be feasible. Immediate postoperative organ function was satisfactory in all patients.

[Abnormal x-ray finding after central venous catheterization]. / Radiologisch auffälliger Befund nach Anlage eines zentralen Venenkatheters.

Central venous catheter placement can cause a variety of complications, such as catheter fracture, loss of the guide wire and embolization. In the case reported a large bore central venous catheter was used in a 32-year-old patient undergoing surgery for vertebral body fracture of the thoracic spine. After a complication-free surgical procedure the post-operative x-ray showed an abnormal finding. A piece of the guide wire was suspected to have been left in the patient. However, this possibility could be ruled out by the anesthesiologist who inserted the catheter. With an additional x-ray and CT scan of an identical catheter it could then be demonstrated that the abnormal finding was caused by polyurethane pins which are integrated in the catheter.

Localization of hepatitis C virus RNA on human erythrocytes by RT in situ PCR technique.

BACKGROUND: Our previous results showed that hepatitis C virus (HCV) is detectable on erythrocytes by RT in situ PCR. The aims of the present study were to compare the sensitivity of this erythrocyte in situ PCR to routine serum solution phase HCV PCR as well as to obtain more data about the binding and cellular localization of HCV in the erythrocyte. METHODS: 105 previously HCV-infected patients and 20 control individuals were studied using RT in situ PCR on erythrocytes and solution phase RT-PCR from serum samples. Binding of HCV to erythrocytes was studied by in vitro inoculation. RT in situ PCR results were evaluated by fluorescence and confocal laser scanning microscopy. RESULTS: From 105 HCV cases, 78 gave positive, while 5-and all control cases-gave negative results by both PCR techniques. In 21 cases, only the in situ technique provided positive results, while in only I case did the solution phase method provide positive results. During in vitro inoculation, an early HCV-erythrocyte binding was detected followed by virus internalization. CONCLUSIONS: Erythrocyte-in situ PCR was found to show higher sensitivity for the detection of HCV compared to the generally applied serum PCR method. In vitro studies suggested a specific binding of HCV to erythrocyte and showed the virus to be capable of internalization.

Hepatitis viruses and hepatocarcinogenesis.

Hepatocellular carcinoma (HCC) is among the most frequent malignancies worldwide. Hepatitis viruses, such as the hepatitis B virus (HBV) and hepatitis C virus (HCV) are undoubtedly listed in the etiology of HCC. Studies show that, in the near future, viral hepatitis will carry increasing weight in the etiology of HCC. This review briefly discusses the known carcinogenic effects of HBV and HCV in the light of experimental and human studies. The data show that viral proteins may directly interfere with gene products responsible for cell proliferation and cell growth. Many other signal transduction cascades may be affected as well. Direct integration of HBV viral sequences into the host genome increases the genomic instability. The genomic imbalance allows the development and survival of malignant clones bearing defected genomic information. HBV and HCV infection induces indirect and direct mechanisms through cellular damage, increased regeneration and cell proliferation, therefore enhancing the development of HCC.

Expression of transforming growth factor-alpha in hepatoblastoma.

BACKGROUND: Transforming growth factor-alpha (TGF-alpha) is a potent stimulator of cell proliferation in the liver and in liver tumors; however, its significance and association with hepatocyte proliferation remains unclear. METHODS: Expression of TGF-alpha and proliferation markers, such as proliferating cell nuclear antigen (PCNA) and cyclin A, were studied and correlated with each other in samples of tumor and surrounding liver tissue taken from nine patients with hepatoblastoma. An avidin-biotin-peroxidase immunohistochemical method was used for detection of TGF-alpha, PCNA, and cyclin A, and in situ hybridization was used to detect TGF-alpha mRNA. RESULTS: Two types of tumor cells of epithelial origin were distinguished based on the expression of TGF-alpha protein and RNA. The more differentiated "fetal" phenotype had a high expression of TGF-alpha and correlated with a low expression of proliferation markers. The less differentiated "embryonal" phenotype had low TGF-alpha expression and high proliferation activity. CONCLUSIONS: The expression of TGF-alpha is associated with a certain morphologic phenotype of tumor cells in hepatoblastoma; higher expression can be detected in more differentiated tumor cells. The negative correlation between the expression of TGF-alpha and proliferation markers suggests that the less differentiated embryonal cells do not depend on growth stimulation provided by TGF-alpha.

[Pathology and pathogenesis of viral hepatitis]. / A vírushepatitisek patológiája és patogenezise.

Several factors viral genotype, mutations, vírus-host interaction, expression of viral proteins and host immune-reaction are very important in the pathogenesis of hepatotrop viral infections. Activation of Fas/Fas ligand system occurs during hepatitis which is responsible for the apoptosis of vírus infected hepatocytes. The histological features in chronic hepatitis can be classified based on the activity of near inflammatory alterations ("grade"). Stage of chronic hepatitis describes the degree of fibrosis and architectural alterations.

[Pilot project for evaluation of lung function reference values]. / Pilotprojekt zur Uberprüfung von Lungenfunktionssollwerten.

Lung function tests were performed in this study on 139 adults (mean age 36 years), as well as on 91 female persons between 16 and 18 years of age (the latter just starting on their professional careers), the lungs being healthy in each case in both groups. The findings were compared with the currently accepted reference value formulae after Quanjer et al., and Zapletal as well as further developed reference value formulae after Brändli et al., Most of the measurement data obtained by spirometry for the BGFA group of probands are between the two recommended reference values for adults with better coincidence with Brändli's predictions (exceptions being MMEF25-75 MEF 50 and MEF 25). In our studies we obtained higher values than the reference median values after Quanjer and Zapletal for IVC, FVC, FEV1 and PEF by 6-8% and 5-15%, respectively, whereas the values for adults differ from the predictions made by Brändli et al. by -4% to +5%. The flow data MMEF 75-25, MEF 50 and especially MEF 25 are set at too low levels (by 5-23%) by Zapletal's and Brändli's values. Comparatively, the values predicted by Quanjer et al. for the above mentioned flow-volume parameters (with the exception of MEF 75) are too high by 4% to 12%. There are also considerable differences in respect of the reference values for IGV to the tune of +15% in the BGFA group compared to Quanjer et al.; in the BAFAM group the values differ from those of Zapletal et al. by +17%. RV yields results in the BGFA group which are higher by 11% than according to Quanjer's formulae, whereas in the BAFAM group they are higher by 15% compared to Zapletal's predictions. In respect of Rt there are differences to the predictions by Rühle and Matthys by +16% (BGFA group) and +13% (BAFAM group), respectively. The BAFAM group differs from Zapletal's predictions by +11%. Looking at the reference limit values the overall impression is confirmed that the predictions after Quanjer et al. and mostly also those by Zapletal are too low in respect of the abovementioned lung function parameters (in the majority of cases not 5% of the examined probands, as expected, are below those levels, but only about 2%). Females, who had been underrepresented in the previous healthy proband groups, show larger deviations than males in respect of most of the parameters.(ABSTRACT TRUNCATED AT 400 WORDS)

[Obligatory intraoperative cholangiography in laparoscopic cholecystectomy]. / Die obligatorische intraoperative Cholangiographie bei der laparoskopischen Cholecystektomie.

Despite growing experience with laparoscopic cholecystectomy in up to now 1100 operations lesions of the bile ducts sometimes occur. We therefore decided to perform intraoperative cholangiography obligatorily and increased our rate of intraoperative X-ray control from 30% in the first 500 operations to 98.2% in the last 500 operations. The mean operation time in the radiography group was 44.8 min. After introduction of intraoperative cholangiography no bile duct lesions were encountered, but in 4.6% of all patients with this examination previously unknown choledocholithiasis was diagnosed.

[Results of conventional and laparoscopic cholecystectomy]. / Ergebnisse der konventionellen und laparoskopischen Cholezystektomie.

We report about a retrospective study of 861 conventional and 812 laparoscopic cholecystectomies (including one coelioscopic choledocholithotomy). In the conventionally operated group reoperation was required in 2.7% (1.4% relaparotomy, 1.3% secondary suture), mortality was 0.5%. After laparoscopic treatment the reoperation rate was 2% (10 relaparoscopies, 5 laparotomies, 2 secondary sutures), no mortality. Growing experience and better definition of contraindications for endoscopic cholecystectomy might improve our results in the future.

[Determination of total lung capacity from the thoracic roentgen image]. / Zur Bestimmung der totalen Lungenkapazität aus Thoraxröntgenaufnahmen.

Formulas for calculation of total lung capacity from chest X-rays were compiled from literature. Using the total lung capacity measured by bodyplethysmography as reference, these formulas and several modifications were tested for usefulness. The product of lung area in the posterior-anterior X-rays and the largest diameter from sternum to the dorsal rib bow in the frontal X-rays was the most simple and exact roentgenological indicator of total lung capacity. 95%-confidence intervals for predictive values gained by this method revealed to be of similar range as for total lung capacity estimated by the gas dilution methods in comparison to bodyplethysmography.

Serological response of experimental ponies orally infected with Ehrlichia risticii.

Ten healthy, mature ponies were orally infected with Ehrlichia risticii using Ehrlichia-infected P388D1 mouse monocyte tissue culture cells. Seven developed signs of equine ehrlichial colitis including fever, depression, anorexia, reduced borborygmi, increased abdominal hyperresonance, and diarrhoea. Three remained clinically normal apart from early fever in one. Indirect fluorescent antibody titres were detected in the clinically affected ponies by Days 12 to 17 post infection and increased rapidly to high levels (1:640 to 1:5120) which were maintained until the end of the observation period (Day 28 post infection). Mean antibody titres significantly increased from Days 12 to 14 and from Days 14 to 17 post infection. The rapid increase in antibody titre which occurs close to first appearance of clinical signs may explain the insignificant rise in antibody titres in many naturally occurring cases.

Intra-abdominal bleeding after rupture of hepatic cyst.

We have described the successful surgical management of intra-abdominal bleeding after rupture of a hepatic cyst. In cases of acute arterial hemorrhage, operation is the treatment of choice, whereas unruptured hepatic cysts seldom cause symptoms and rarely need treatment.

[Incidence of complications in elective colon resections at various ages]. / Komplikationsdichte elektiver Kolonresektionen in verschiedenen Lebensaltern.

A retrospective analysis was made of 235 cases of colectomy, subdivided by age groups, with emphasis being laid on preoperative risk factors and postoperative complications. Lethality accounted for 2.5 per cent, with disorders of wound healing being the most common complications. Advanced age per se does not imply unfavourable prognosis.

Iron uptake by rat duodenal microvillous membrane vesicles: evidence for a carrier mediated transport system.

The mechanism of iron translocation from intestinal lumen to portal plasma is poorly understood. To examine these processes, uptake of Fe2+ and Fe3+ by rat duodenal microvillous membrane vesicles prepared by a Ca2+ precipitation procedure was studied. Membrane aliquots were incubated with increasing concentrations of 59FeCl3 in the presence of a one-thousand-fold molar excess of citrate or 59FeSO4 with a twenty-fold molar excess of L-ascorbic acid. After various time intervals the incubation reaction was stopped by addition of 0.1 mM FeCl3 (4 degrees C), and uptake of 59Fe was determined by a vacuum filtration assay. Initial uptake velocity of 59FeCl3 and 59FeSO4 was determined from the slope of the cumulative uptake curves, which was linear for the first 60 s. Initial uptake rates of both, 59Fe3+ and 59Fe2+ revealed an identical saturable uptake component with a Km of 19-22 nM and Vmax of 8 pmol min-1 mg protein-1. In addition, transport of Fe2+ revealed a linear unspecific uptake phase, which was predominant at high substrate concentrations. Saturable uptake of Fe2+ and Fe3+ was temperature dependent, and significantly reduced by trypsin pretreatment of the microvillous membrane vesicles, indicating the involvement of a protein in the uptake process. This suggestion was pursued by isolation of an iron binding protein from duodenal brushborder membranes. After solubilization of microvillous plasma membranes with 1% Triton X 100, affinity chromatography of the membrane protein mixture over an iron chelate gel derived from epoxy activated Sepharose and elution with 50 mM EDTA yielded a single 52,000 dalton protein. The protein co-chromatographed over an Ultro-Pac TSK G 3000SW HPLC column together with 59FeCl3 and 59FeSO4. It showed no immunologic activity to rabbit antibodies against whole rat serum or rat transferrin. Furthermore, by photoaffinity labelling technique a single iron binding protein with a molecular weight of about 52,000 dalton was identified in microvillous membranes of the rat duodenum. These data are compatible with the hypothesis that intestinal iron absorption is mediated by a specific carrier-dependent transport system.