- Email: [email protected]
Localized tissue destruction by hepus in pigs
A536
AGA ABSTRACTS
GASTROENTEROLOGY, Vol. 108, No. 4
• EFFECTS OF HIGH ENERGY PULSED ULTRASOUND ON NORMAL AND MALIGNANT LIVER CELLS. H.T. Schneider, T. Feigl, B.A. Vrlklein, R. Riedlinger*, E.G. Hahn, C. Ell. Depts. of Medicine I, University of Erlangen-Nuremberg and *Acoustics IHE, University of Karlsruhe, Germany
• LOCALIZED TISSUE DESTRUCTION BY HEPUS IN PIGS. H.T. Schneider, T. Feigl, R. Riedlinger'*, H. Schubert*'*, J. Hornung', Ch. Wittekind °, E.G. Hahn, C. Ell. Depts. of Medicine I and "Pathology, University of Erlangen-Nuremberg, and "'Acoustics IHE, University of Karlsruhe, and "'IVTK, University of Jena, Germany
Recently, we have shown that pancreatic tumor cells can be successfully destroyed by high energy pulsed ultrasound (HEPUS)(Eur J Gastroenterol & Hepatol 1994). In order to evaluate the sensitivity to HEPUS depending on cell differentiation, we now compared the effects on human benigne and malignant cells. Methods: Normal liver cells (Chang Liver) and liver carcinoma cells (SKHEP-1) were trypsinized, concentrated (0.5-2 x 106 cells/ml) and aliquots (2 ml) transferred to polyethylene tubes. They were exposed to HEPUS using an experimental piezoelectric transducer characterized by a high negative peak pressure amplitude. Cell survival rates were determined depending on the number of applied pulses, the voltage of the transducer and the pulse repetition frequency (Hz) by means of trypan blue dye exclusion :test and detection of the lactate dehydrogenase CLDH) in the supematent. '.Additionally, ultrastructural changes of the exposed cells were investigated :by means of electron microscopy. Results: Both, tumor cells and non-malignant cells showed an exponentially decreasing number of surviving cells by increasing the number of pulses (0-800/2000). Increasing the applied voltage (40-100% Umax) resulted in a linear decrease of surviving cells. Within the range of 0.6-8 ~Hz maximum destruction rate was observed at 1 Hz pulse repetition frequency. To reach a level of 99% cell destruction of the tumor and the normal cells 400 pulses and 2000 pulses had to be administred respectively. Applying 400 pulses at 90% of the maximum voltage resulted in a survival rate of less than 5% of the tumor cells compared to 30% of the non-malignant ceils. Measurements of the LDH confirmed the data obtained by the cell counts. Electron microscopy revealed changes in the cellular structure such as vacuoles, localized ruptures and cell fragmentation, quantitatively and qualitatively depending on the applied number of pulses. Conclusions: Malignant and non-malignant ceils of the liver showed significant dependencies on the applied number of pulses, puls repetition rate and generator voltage. As opposed to the normal ceils, the malignant liver cells proved to be more sensitive to HEPUS exposure.
Experiments in vitro (tumor ceils) and in vivo (tumor bearing nude mice) showed the capability of high energy pulsed ultrasound (I-IEPUS) to destroy suspended tumor ceils as well as tumor tissue. Prior to a possible application of HEPUS in tumor patients, the potential of selectively destroying deeper tissue layers without affecting surrounding tissue needed to be verified. Methods: Eight domestic pigs (29-2z7kg) were anesthetized and the liver exposed to HEPUS, using an experimental piezoelectric transducer characterized by a high negative peak pressure amplitude in the burst signal to generate microcavitation (focal area at 50% isobar: x,y,z: 5x5x25mm). Localizing of the tissue was achieved by an integrated ultrasound scanner (3.5/5MHz). In all animals a predetermined cubic volume of 12x12x12mm was sonicated at 4x4x2 points administering 200 pulses per point. The focal depth in the bodie s of.the different pigs varied from 54mm to 97mm from skin. Two animals were sacrificed within eight hours, six animals eight days after the treatment. The liver was resected, macroscopically and histologically (H&E staining) examined, Results: HEPUS application was well tolerated by all animals. During sonication cavitation could be sonographically observed as hyperechogenic areas around the focus. In all pigs the HEPUS-treated tissue areas of the liver were visible at resection. Eight hours after treatment the lesions appeared macmscopically as hemorrhagic areas; microscopy showed tissue disruption with hemorrhages and necrosis of the liver cells and a sharp boundary to the non-treated tissue. Eight days after treatment scared and shrinked tissue was seen macroscopically at the expected location. Histologic evaluation demonstrated fibrotic walls around necrotic cavities with infiltrates of lymphocytes and plasma cells. The lesions were sharply demarcated to the surrounding liver. Conclusions: HEPUS induces targeted tissue destruction under sonographic monitoring even in deeper tissue layers of the body by generating microcavitation. The lesions revealed sharp boundaries without microscopic visible alterations of the surrounding tissue. The present equipment seems to be applicable in patients with the aim to destroy tumors palliatively.
PROGNOSTIC VALUE OF THE EXPRESSION OF DI,6 BRANCHED OLIGOSACCHARIDES IN HUMAN COLON CARCINOMAS W.K.F. Seelentag*, W.-P. Li*, Ch. Zuber*, B. Stature#, Ph.U. Heitz*, J. Roth*.* Div. of Cell and Molecular Pathology, Dept. of Pathology. University of Ztirich and #Institute for Pathology, Cantonal Hospital Aarau; Switzerland A positive correlation between the metastatic potential of tumor cells and increased 131,6 branching in oligosaccharides has been recentl,~ demonstrated. Previously we have shown that such a correlation can be extended to variants of metastasizing human colon carcinoma HCT116 cells. In this study we examined if the expression of lectin-histochemicall~ detected 131,6 branched oligosaccharides is correlated (i) to the Dukes stage as modified by Astler an Coller and (ii) to the clinical course of colon carcinoma. In colon carcinoma, the intensity and pattern of staining tbr 131,6 branches did not show a correlation (by analysis of variance) with the Dukes stage. In the total patient collective, there was a slight, however statisticalb not significant, decrease of the staining intensity with increasing survival time (follow up until 11 years) and increasing disease free interval. Also within different sub-groups according to the Dukes stage or to the degree of differentiation of the tumor, there was no statistically significant correlation between the staining intensity and the disease free interval. The histochemically demonstrated expression of 131,6 branches in surgical specimens does not significantly correlate with the clinical outcome, in contrast to previously examined experimental systems Where aggressiveness of human colon carcinoma cell lines was correlated with qualitative and quantitative changes of 131,6 branched oligosaccharides.
• FOODS AS RISK FACTORS FOR COLORECTAL CANCER : A CASECONTROL STUDY IN A FRENCH AREAl P. Senesse, M.C. Boutron, I. Faivre. Registre Bourguignon des Cancers Digestifs, Facult6 de Mrdecine, 21000 Dijon, and INSERM U290, 75010 Paris, France. Epid~aiologlcal study have strongly suggestedthat a high saturated fatlow fibre, low carbohydrate diet was associated with an increased risk of colorectal cancer. However, this had been mostly demonstrated in North American or anglo-saxoncountries where diet is very different from the diet of Latin European countries. We compared diet in colorectal cancer cases (n= 171) with diet of general population controls (n=309) in a well defined French area. Consumptions were classified into quintiles, or quartiles if the food was less frequently consumed, according to the distribution in the control group, and separately by sex. Relative risks were computed with a logistic regression controlling for age and caloric intake. Odds ratios (OR) are given for the highest versus lowest category of intake ; p values (p) correspond to the test for linear trend. Considering widely consumed foods, risk of cancer was significantly associated with high intakes of reiined cereal (OR=2.2; p=0.002), or office and pasta (OR=2:1; p=0.04). It was also significantly associated with processed meat and delicatessen (OR=3.0; p=0.001) and particularly patrs and spread (OR=2.4; p=0.1) ; no significant association was observed with fresh meat (OR=I. 1; p=0.9). Regarding fat intake (fats added to foods), it was significantly associated with risk of cancer (OR=2.0; p=0.007), and, when considering type of fat, the strongest association was found for animal fat (lard, butter and cream) with an OR for the fourth vs first quartile of intake of 2.1; p=0.06. No significantly protective factor could be determined, although there was a trend towards a protective effect ofvegctahles (OR=0.7; p=0.2). There was no protective effect of dairy products (OR=0.9; p=0.4). These data suggest that, in a french population, a diet rich in animal fat, in processed meat and parrs, and in refined cereal, and low in vegetables could be associated with an increased risk ofcolorectal cancer. They do not support the hypothesis of a protective effect of milk and dairy products nor of an increased risk with high consumptions of fresh meat. Such data demonstrate that the relative importance of dietary factors associated with risk of colorectal cancer may differ from one country to another ; this should be taken into account when giving dietary advice to the population.
AGA ABSTRACTS
GASTROENTEROLOGY, Vol. 108, No. 4
• EFFECTS OF HIGH ENERGY PULSED ULTRASOUND ON NORMAL AND MALIGNANT LIVER CELLS. H.T. Schneider, T. Feigl, B.A. Vrlklein, R. Riedlinger*, E.G. Hahn, C. Ell. Depts. of Medicine I, University of Erlangen-Nuremberg and *Acoustics IHE, University of Karlsruhe, Germany
• LOCALIZED TISSUE DESTRUCTION BY HEPUS IN PIGS. H.T. Schneider, T. Feigl, R. Riedlinger'*, H. Schubert*'*, J. Hornung', Ch. Wittekind °, E.G. Hahn, C. Ell. Depts. of Medicine I and "Pathology, University of Erlangen-Nuremberg, and "'Acoustics IHE, University of Karlsruhe, and "'IVTK, University of Jena, Germany
Recently, we have shown that pancreatic tumor cells can be successfully destroyed by high energy pulsed ultrasound (HEPUS)(Eur J Gastroenterol & Hepatol 1994). In order to evaluate the sensitivity to HEPUS depending on cell differentiation, we now compared the effects on human benigne and malignant cells. Methods: Normal liver cells (Chang Liver) and liver carcinoma cells (SKHEP-1) were trypsinized, concentrated (0.5-2 x 106 cells/ml) and aliquots (2 ml) transferred to polyethylene tubes. They were exposed to HEPUS using an experimental piezoelectric transducer characterized by a high negative peak pressure amplitude. Cell survival rates were determined depending on the number of applied pulses, the voltage of the transducer and the pulse repetition frequency (Hz) by means of trypan blue dye exclusion :test and detection of the lactate dehydrogenase CLDH) in the supematent. '.Additionally, ultrastructural changes of the exposed cells were investigated :by means of electron microscopy. Results: Both, tumor cells and non-malignant cells showed an exponentially decreasing number of surviving cells by increasing the number of pulses (0-800/2000). Increasing the applied voltage (40-100% Umax) resulted in a linear decrease of surviving cells. Within the range of 0.6-8 ~Hz maximum destruction rate was observed at 1 Hz pulse repetition frequency. To reach a level of 99% cell destruction of the tumor and the normal cells 400 pulses and 2000 pulses had to be administred respectively. Applying 400 pulses at 90% of the maximum voltage resulted in a survival rate of less than 5% of the tumor cells compared to 30% of the non-malignant ceils. Measurements of the LDH confirmed the data obtained by the cell counts. Electron microscopy revealed changes in the cellular structure such as vacuoles, localized ruptures and cell fragmentation, quantitatively and qualitatively depending on the applied number of pulses. Conclusions: Malignant and non-malignant ceils of the liver showed significant dependencies on the applied number of pulses, puls repetition rate and generator voltage. As opposed to the normal ceils, the malignant liver cells proved to be more sensitive to HEPUS exposure.
Experiments in vitro (tumor ceils) and in vivo (tumor bearing nude mice) showed the capability of high energy pulsed ultrasound (I-IEPUS) to destroy suspended tumor ceils as well as tumor tissue. Prior to a possible application of HEPUS in tumor patients, the potential of selectively destroying deeper tissue layers without affecting surrounding tissue needed to be verified. Methods: Eight domestic pigs (29-2z7kg) were anesthetized and the liver exposed to HEPUS, using an experimental piezoelectric transducer characterized by a high negative peak pressure amplitude in the burst signal to generate microcavitation (focal area at 50% isobar: x,y,z: 5x5x25mm). Localizing of the tissue was achieved by an integrated ultrasound scanner (3.5/5MHz). In all animals a predetermined cubic volume of 12x12x12mm was sonicated at 4x4x2 points administering 200 pulses per point. The focal depth in the bodie s of.the different pigs varied from 54mm to 97mm from skin. Two animals were sacrificed within eight hours, six animals eight days after the treatment. The liver was resected, macroscopically and histologically (H&E staining) examined, Results: HEPUS application was well tolerated by all animals. During sonication cavitation could be sonographically observed as hyperechogenic areas around the focus. In all pigs the HEPUS-treated tissue areas of the liver were visible at resection. Eight hours after treatment the lesions appeared macmscopically as hemorrhagic areas; microscopy showed tissue disruption with hemorrhages and necrosis of the liver cells and a sharp boundary to the non-treated tissue. Eight days after treatment scared and shrinked tissue was seen macroscopically at the expected location. Histologic evaluation demonstrated fibrotic walls around necrotic cavities with infiltrates of lymphocytes and plasma cells. The lesions were sharply demarcated to the surrounding liver. Conclusions: HEPUS induces targeted tissue destruction under sonographic monitoring even in deeper tissue layers of the body by generating microcavitation. The lesions revealed sharp boundaries without microscopic visible alterations of the surrounding tissue. The present equipment seems to be applicable in patients with the aim to destroy tumors palliatively.
PROGNOSTIC VALUE OF THE EXPRESSION OF DI,6 BRANCHED OLIGOSACCHARIDES IN HUMAN COLON CARCINOMAS W.K.F. Seelentag*, W.-P. Li*, Ch. Zuber*, B. Stature#, Ph.U. Heitz*, J. Roth*.* Div. of Cell and Molecular Pathology, Dept. of Pathology. University of Ztirich and #Institute for Pathology, Cantonal Hospital Aarau; Switzerland A positive correlation between the metastatic potential of tumor cells and increased 131,6 branching in oligosaccharides has been recentl,~ demonstrated. Previously we have shown that such a correlation can be extended to variants of metastasizing human colon carcinoma HCT116 cells. In this study we examined if the expression of lectin-histochemicall~ detected 131,6 branched oligosaccharides is correlated (i) to the Dukes stage as modified by Astler an Coller and (ii) to the clinical course of colon carcinoma. In colon carcinoma, the intensity and pattern of staining tbr 131,6 branches did not show a correlation (by analysis of variance) with the Dukes stage. In the total patient collective, there was a slight, however statisticalb not significant, decrease of the staining intensity with increasing survival time (follow up until 11 years) and increasing disease free interval. Also within different sub-groups according to the Dukes stage or to the degree of differentiation of the tumor, there was no statistically significant correlation between the staining intensity and the disease free interval. The histochemically demonstrated expression of 131,6 branches in surgical specimens does not significantly correlate with the clinical outcome, in contrast to previously examined experimental systems Where aggressiveness of human colon carcinoma cell lines was correlated with qualitative and quantitative changes of 131,6 branched oligosaccharides.
• FOODS AS RISK FACTORS FOR COLORECTAL CANCER : A CASECONTROL STUDY IN A FRENCH AREAl P. Senesse, M.C. Boutron, I. Faivre. Registre Bourguignon des Cancers Digestifs, Facult6 de Mrdecine, 21000 Dijon, and INSERM U290, 75010 Paris, France. Epid~aiologlcal study have strongly suggestedthat a high saturated fatlow fibre, low carbohydrate diet was associated with an increased risk of colorectal cancer. However, this had been mostly demonstrated in North American or anglo-saxoncountries where diet is very different from the diet of Latin European countries. We compared diet in colorectal cancer cases (n= 171) with diet of general population controls (n=309) in a well defined French area. Consumptions were classified into quintiles, or quartiles if the food was less frequently consumed, according to the distribution in the control group, and separately by sex. Relative risks were computed with a logistic regression controlling for age and caloric intake. Odds ratios (OR) are given for the highest versus lowest category of intake ; p values (p) correspond to the test for linear trend. Considering widely consumed foods, risk of cancer was significantly associated with high intakes of reiined cereal (OR=2.2; p=0.002), or office and pasta (OR=2:1; p=0.04). It was also significantly associated with processed meat and delicatessen (OR=3.0; p=0.001) and particularly patrs and spread (OR=2.4; p=0.1) ; no significant association was observed with fresh meat (OR=I. 1; p=0.9). Regarding fat intake (fats added to foods), it was significantly associated with risk of cancer (OR=2.0; p=0.007), and, when considering type of fat, the strongest association was found for animal fat (lard, butter and cream) with an OR for the fourth vs first quartile of intake of 2.1; p=0.06. No significantly protective factor could be determined, although there was a trend towards a protective effect ofvegctahles (OR=0.7; p=0.2). There was no protective effect of dairy products (OR=0.9; p=0.4). These data suggest that, in a french population, a diet rich in animal fat, in processed meat and parrs, and in refined cereal, and low in vegetables could be associated with an increased risk ofcolorectal cancer. They do not support the hypothesis of a protective effect of milk and dairy products nor of an increased risk with high consumptions of fresh meat. Such data demonstrate that the relative importance of dietary factors associated with risk of colorectal cancer may differ from one country to another ; this should be taken into account when giving dietary advice to the population.