Ferenc Jakab

Comparison of Human Hepatoma HepaRG Cells with Human and Rat Hepatocytes in Uptake Transport Assays in Order to Predict a Risk of Drug Induced Hepatotoxicity

Human hepatocytes are the gold standard for toxicological studies but they have several drawbacks, like scarce availability, high inter-individual variability, a short lifetime, which limits their applicability. The aim of our investigations was to determine, whether HepaRG cells could replace human hepatocytes in uptake experiments for toxicity studies. HepaRG is a hepatoma cell line with most hepatic functions, including a considerable expression of uptake transporters in contrast to other hepatic immortalized cell lines. We compared the effect of cholestatic drugs (bosentan, cyclosporinA, troglitazone,) and bromosulfophthalein on the uptake of taurocholate and estrone-3-sulfate in human and rat hepatocytes and HepaRG cells. The substrate uptake was significantly slower in HepaRG cells than in human hepatocytes, still, in the presence of drugs we observed a concentration dependent decrease in uptake. In all cell types, the culture time had a significant impact not only on the uptake process but on the inhibitory effect of drugs too. The most significant drug effect was measured at 4 h after seeding. Our report is among the first concerning interactions of the uptake transporters in the HepaRG, at the functional level. Results of the present study clearly show that concerning the inhibition of taurocholate uptake by cholestatic drugs, HepaRG cells are closer to human hepatocytes than rat hepatocytes. In conclusion, we demonstrated that HepaRG cells may provide a suitable tool for hepatic uptake studies.

Inflammation of ectopic pancreatic tissue in a Meckel's diverticulum causing acute abdominal symptoms: a case report and review of the literature.

An unusual case of acute abdomen was caused by the inflammation of ectopic pancreatic tissue in a Meckel’s diverticulum. A 49-year-old man presented with acute abdominal pain, and the clinical diagnosis of acute appendicitis was established. During laparotomy, a normal appendix of unusual localization near the gallbladder and a Meckel’s diverticulum with an inflamed tip were found. Histological examination showed acute inflammation of heterotopic pancreatic tissue along with normal ectopic gastric and duodenal mucosa within the wall of the diverticulum. Fat necrosis was also ascertained. The authors believe that this is the first report of acute inflammation of ectopic pancreatic tissue and the presence of normal ectopic gastric and duodenal tissue in the same Meckel’s diverticulum.

Differential inhibitory effect of cyclosporin A and bosentan on taurocholate uptake in human and rat hepatocytes as a function of culturing time

Bile salt transport across hepatocytes requires a coordinate action of transporters, which is thought to be a target for drug-induced cholestasis. Hepatocytes provide the most competent in vitro model to predict transporter-related toxic drug effects. The aim of this study was to show a correlation between inhibitory potential of drugs and the change of rate, as well as of the active to passive ratio of taurocholate uptake in these cells. In rat hepatocytes, along with a significant decrease of uptake (86.4% by 72h), and the shift of saturable/unsaturable transport (from 92/8 to 55/45 in a 24-72h time interval), the efficacy of taurocholate uptake inhibition was highly reduced (IC(50) cyclosporin A 3.9 to >100μM, and bosentan 9.1-49.8μM at 1 and 72h, respectively). In contrast, 5-day-old human hepatocytes preserved 70% of their taurocholate uptake capacity with a 2-fold higher active than passive transport, which resulted in a more efficient inhibition by drugs (IC(50) cyclosporin A, 2.4 to ∼10μM and bosentan 28.9-45.5μM at 1h and 5days, respectively). Our results support that reliable drug interaction studies might be performed in 5-day-old human hepatocyte cultures, while experiments using rat hepatocytes at more than 24h after seeding will highly underestimate the probability of drug interaction.

Decreased functional activity of multidrug resistance protein in primary colorectal cancer

BackgroundThe ATP-Binding Cassette (ABC)-transporter MultiDrug Resistance Protein 1 (MDR1) and Multidrug Resistance Related Protein 1 (MRP1) are expressed on the surface of enterocytes, which has led to the belief that these high capacity transporters are responsible for modulating chemosensitvity of colorectal cancer. Several immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) studies have provided controversial results in regards to the expression levels of these two ABC-transporters in colorectal cancer. Our study was designed to determine the yet uninvestigated functional activity of MDR1 and MRP1 transporters in normal human enterocytes compared to colorectal cancer cells from surgical biopsies.Methods100 colorectal cancer and 28 adjacent healthy mucosa samples were obtained by intraoperative surgical sampling. Activity of MDR1 and MRP1 of viable epithelial and cancer cells were determined separately with the modified calcein-assay for multidrug resistance activity and sufficient data of 73 cancer and 11 healthy mucosa was analyzed statistically.ResultsSignificantly decreased mean MDR1 activity was found in primary colorectal cancer samples compared to normal mucosa, while mean MRP1 activity showed no significant change. Functional activity was not affected by gender, age, stage or grade and localization of the tumor.ConclusionWe found lower MDR activity in cancer cells versus adjacent, apparently, healthy control tissue, thus, contrary to general belief, MDR activity seems not to play a major role in primary drug resistance, but might rather explain preferential/selective activity of Irinotecan and/or Oxaliplatin. Still, this picture might be more complex since chemotherapy by itself might alter MDR activity, and furthermore, today limited data is available about MDR activity of cancer stem cells in colorectal cancers.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1675739129145824

Laparoscopic or open appendectomy

UNLABELLED Despite the fact that laparoscopic appendectomy was one of the first performed minimally invasive surgical procedures, the benefits and indications of its use are still controversial. METHODS Data of patients with the diagnosis of appendicitis operated either with laparoscopic (LA) or open (OA) appendectomy were collected and analyzed. PATIENTS This study involved 273 consecutive patients who had undergone appendectomy with the diagnosis of acute appendicitis at the Uzsoki Teaching Hospital, Department of Surgery between January 2005 and December 2007. 185 patients (68%) operated with the laparoscopic, 88 (32%) with the open technique, in the third year 89% of the cases underwent laparoscopic appendectomy. RESULTS The conversion rate was 27%, the reason of the conversion was the progression of the disease in 35 patients (70%) and technical in 15 cases (30%). The wound infection rate was 8% in the LA and 18% in the OA group ( p = 0.022). No significant difference was found in the reoperation rate and in the hospital readmission rate between the two groups. There was one insufficiency which was treated with conservative therapy. The hospital stay was significantly lower in the laparoscopic group ( p = 0.031). CONCLUSIONS This study demonstrated that laparoscopic appendectomy has significant advantages over open appendectomy. In our practice, laparoscopic appendectomy is the first choice of procedure in acute appendicitis.

Ismételt májresectio, a modern daganatsebészet egyik kihívásar

Surgical and oncological treatment of liver metastases of CRC has made a significant progress in the last twenty years. Although chemotherapy has developed enormously, only successfully resected (R0) patients may hope a long-term survival. The almost 60% recurrence rate after resection is a significant challenge. Authors review patients data operated at the Surgical Department of Uzsoki Teaching Hospital between 1995 and 2008 with hepatic metastases of colorectal origin. Oncological and surgical principles are summarized in the sight of the recent literature review and authors own experience with repeat hepatic resection. Recent literature data demonstrate--which is confirmed by our experience--that R0 resection of recurrent hepatic metastases provides similar overall 5 year survival rate than that of patients who underwent a single resection only. In the case of recurrent CLM, the oncoteam should prepare a surgically resectable situation.

From project based sample collection to a biobank

The research group takes samples for molecular genetical examinations from tumors removed during operations within ischemic time interval. Samples are stored in liquid nitrogen. Clinical data of these patients are recorded in an informatics system developed by the group. Patients are followed in an out-patient clinic set up for this purpose not financed by the National Health Insurance Fund. Tissue samples and follow up data are used to cooperate with molecular genetical laboratories.

Multiple hepatic abscesses due to actinomycosis: difficulties of diagnosis

Authors present the case of a 65 year-old man without any remarkable previous illness, hospitalized because of having prolonged fever caused probably by an infection due to a tick bite. Tularaemia and bartonellosis based on serological results as well as neoplasm of the sigma with multiple metastases to the liver raised by ultrasonography and abdominal CT could be excluded with detailed investigations. Multiple abscesses of the liver were diagnosed, followed by a percutaneuos ultrasonography directed aspiration of one of the abscesses which resulted in 150 ml buffy pus. Microbiological examination evaluated the presence of Actinomyces meyeri. Instead of neoplasm, diverticulosis and diverticulitis of the sigma could be clarified with perforation of one of the diverticuli causing peridiverticulitis, retroperitoneal abscess and probably abscesses in the liver by haematogenous dissemination. Surgical intervention, resection of the liver and the sigma resulted in recovery of the patient. Difficulties of diagnosis are discussed in the article pointing out the importance of interdisciplinary collaboration, involving also infectologist. Diverticulosis of the sigma as presumably origin of hepatic actinomycosis is a literary curiosity. This was the cause of the case presentation.

[Choice of surgical strategy for colorectal liver metastases depending on clinical and pathological response to neoadjuvant chemo- and targeted therapy].

UNLABELLED Surgical strategy of colorectal liver metastases depends on clinical and pathological response to neoadjuvant chemo- and target therapy. Recently, surgical treatment of advanced colorectal cancer appears as an everyday challenge for surgeons and oncologists. The new oncologic procedures invented last years led to significant therapeutic improvement. Combination of neoadjuvant chemotherapy with biological answer modifiers increased greatly the clinical response rate given to cytoreductive therapy. Due to these facts the complete disappearance of liver metastases can be observed more and more frequently after 2000. Literature of metastasis surgery clarified the exact difference between clinical and pathological response, at the same time exact criteria of the connections between two responses have been set. In complex treatment of colorectal liver metastases complete pathological response became the end point of treatment. AIMS AND METHODS Between January, 2009 and August, 2010, 39 patients with originally non resectable liver metastases (colorectal cancer origin) have been studied. All patients were treated by neoadjuvant chemo- and targeted therapy before the resection of liver. RESULTS Complete pathological response has been observed in 3 patients, major pathological response (necrosis: >50%) in 11 patients, minor pathological answer (necrosis <50%) in 22 patients and finally no necrosis at all in 3 patients. CONCLUSIONS Complete pathological response can be regarded as the final goal of neoadjuvant targeted therapy. Pathological response seems to be the most important prognostic factor which reflects long-term survival after the R0 resection of liver metastases. Complete disappearance of liver metastases is an undesirable side effect of oncological therapy, which causes difficulties during surgical intervention. In this term the overtreatment of patients resulting in a disappearance of metastases should be avoided. Multidisciplinary team is responsible for the indication of resection of liver metastases in time before their disappearance.

Surgical strategy of colorectal liver metastases treatment depending on clinical and pathological response to neoadjuvant chemo- and target therapy

UNLABELLED Surgical strategy of colorectal liver metastases depends on clinical and pathological response to neoadjuvant chemo- and target therapy. Recently, surgical treatment of advanced colorectal cancer appears as an everyday challenge for surgeons and oncologists. The new oncologic procedures invented last years led to significant therapeutic improvement. Combination of neoadjuvant chemotherapy with biological answer modifiers increased greatly the clinical response rate given to cytoreductive therapy. Due to these facts the complete disappearance of liver metastases can be observed more and more frequently after 2000. Literature of metastasis surgery clarified the exact difference between clinical and pathological response, at the same time exact criteria of the connections between two responses have been set. In complex treatment of colorectal liver metastases complete pathological response became the end point of treatment. AIMS AND METHODS Between January, 2009 and August, 2010, 39 patients with originally non resectable liver metastases (colorectal cancer origin) have been studied. All patients were treated by neoadjuvant chemo- and targeted therapy before the resection of liver. RESULTS Complete pathological response has been observed in 3 patients, major pathological response (necrosis: >50%) in 11 patients, minor pathological answer (necrosis <50%) in 22 patients and finally no necrosis at all in 3 patients. CONCLUSIONS Complete pathological response can be regarded as the final goal of neoadjuvant targeted therapy. Pathological response seems to be the most important prognostic factor which reflects long-term survival after the R0 resection of liver metastases. Complete disappearance of liver metastases is an undesirable side effect of oncological therapy, which causes difficulties during surgical intervention. In this term the overtreatment of patients resulting in a disappearance of metastases should be avoided. Multidisciplinary team is responsible for the indication of resection of liver metastases in time before their disappearance.