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A longitudinal study of HCV RNA detected by in situ-RT-PCR in liver biopsy specimens during progression from chronic HCV liver disease to hepatocellular cancer, HCC
A494
AGA ABSTRACTS
GASTROENTEROLOGY, VOl. 108, NO. 4
ENDOSCOPIC, L A P A R O S C O P I C A N D OPEN S U R G E R Y FOR
IN P U R S U I T O F
EARLY GASTRIC CANCER. K. Kumai, Y. Otani, M. Ogami, T, Kubota, M. Kitajima. Depts. of Surgery, School of Medicine, Keio University, Tokyo, Japan
A Kumar, Division of Gastroenterology, Univ of Tennessee at Memphis. Completeness of retrieval of studies is imperative for good oncological epidemiology. MEDLINE search does not retrieve a significant number of articles.Beginners do not have access to not easily available [Elusive] literature.We investigated these sources of information. M~ETHOD:Review of available literature SOURCES OF ELUSIVE L I T E R A T U R E - Published works: subject catalogues, abstracts, indexes,reviews, databases [EXCERPTA, EMBASE]. Retrieval continued till authors names appear again [&again].Completed works:Papers/technical reports-National Technical Information Center [NTIS] database, 26 areas, 88% completion, abstract newsletter weekly: Educational Resources Information Center[ERIC] database :Federal Database Finder [Information,USA, 1986]: Monthly catalog of US Government Publications[Office Superintendent Documents] Dissertations/theses-DATRIX II&CDI [Univ Microfilm International, AnnArbor,MI]: Bibliography Index[Oryx Press 1985]MeetingsBibliographic Guide [G K Hall,Boston], Conference Papers Index[Cambridge Scientific Abstracts,Bethesda]Census/economic dataCensus Information Census:Economic Bulletin Board:Population Bibliography:Statistical Analyses [Elveiser1991]Studies in progress: Federal Reseach In Progress[NTIS]:UNIVRES [Canada],CRIB[Britain] and STRC[Australia] {Elveiser1991}:EUDISED[Europe]:FORIS[Germany] Informal means-meetings,electronic networks, public policy organisations,researchers in same field.
The majority of mucosal gastric cancers could be curatively treated by local resection of the stomach without lymph node dissection, because lymph node metastases were observed in only 10/475 patients(2.0%). As there was no lymph node metastasis in the cases who had lesions less than 2.5 cm in diameter in our department, Endoscopic Mucosal Resection (EMR) has been applied for 70 patients with an elevated mucosal lesion less than 2 cm in diameter or with a depressed lesion less than 1 cm, but 5 cases were converted to open surgery technically. Therefore we limited the indication of EMR for a lesion less than 1 cm in diameter. And as a new method, we t r e a t e d l 8 patients by 2 laparoscopic procedures. Laparoscopic wedge resection of the stomach was performed for 9 patients by Lesion Lifting Method. The gastric lesion was lifted laparoscopically using the string which was pierced the gastric wall just near the lesion. Wedge resection w a s c a r r i e d out with a sufficient surgical margin. Laparoscopic Intragastric Mucosal Resection was also performed for 9 cases who had lesions at the posterior wall or near the cardia or the pyloric ring. Open surgery using modified lymph node dissection could be applied for other mucosal cancers. A standard radical lymph node dissection should be need for submucosally invaded cancers, because 13.5% of lymph node metastasis was observed.
K~RAS2 C O D O N 12 G A T AND G T T C O E X I S T I N G SSCPs OCCUR FREQUENTLY IN PERIAMPULLARY N E O P L A S M S OF P A N C R E A T I C AS W E L L AS O F B I L I A R Y ORIGIN. L.Laghi, L. Arborati, O. Orbetegli, M. Sampiotro, A. Zerbi, D. Faravelli, V.Di Carlo, A. Malesci, G. Fiorelli. Istituto di Medicina Intema e Fisiopatologia Medica, Ospedale Policlinico; Clinica Chirurgica e Anatomia Patologica, Istituto S. Raffaele, Milano, Italy. Pancreatic cancer has the highest prevalence of Kras2 codon 12 mutations of any human nooplasm.Transve:'sion to G A T accounts for 50% of mutations, and transversion to G T T for 30%, while other substituitions are reportod with variable frequency. Coexisfiug mutations are rarely (<5%) described. More heterogeneous data are available for bile duct neoplasms, where coexisting mutations (mainly G A T and GTT) occur to varying degrees (30-100%). AIM: We assessed whether periampullary pancreatic and biliary tumors would produce different Single Straod Conformation Polymorphism (SSCP) patterns of Kras2 exonl, and pm'dcularly patterns homologous to G A T aud G ' I T codonl2 mutations. M E T H O D S : 56 specimens of periampullary neoplasms have been processed. Using a heminestod PCR technique, SSCPs of 107bpamplic0ns (spanniug the nt -13 +93 in the first exon of Kras2) were obteined from paraffin-embedded neoplastic tissues and compared with D N A s control with known G A T and G T T mutations. SSCPs were detected by silver staining of 15% PAGE-minigeI employing conditions standardized to detect both mutations simultaneously. RESULTS: SSCP patterns different from normal were detected in 34141(83%) pancreatic ductal neoplasms,0/2 ampollary neoplasms, 4/4 acinar neoplasms and i0/10 distal bile duct. O b s e r v e d SSCP p a t t e r n s T u m o r s type GAT GTI" IGAT+GTT Other Bilim7 0 0 7(70%) " 3(30%) DuctalPancreatic 9(26%) 9 (26%) 12 (35%) 4 (13%) AmpullatT 0 0 0 0 Acinar 1(25%) 0 3 (75%) 0 Coexistence of different SSCP patterns (mainly GAT and GTT-like) in pancreatic ductal-originating tumors was associated with the presence of mucinous areas (7/9 vs 7125, X2=0.025). C O N C L U S I O N S : Our data suggest dmt 1) multiple coexisting Kras cod 12 mutations occur in bile duct neoplasms ; 2) double mutations in ductal-ofiginating tumors of the pancreas are more frequently seen in mucinous areas; 3) also acinar lesions can progress through these genetic changes. Microdissection studies can clarify if it is possible recognize diffelrent areas in neoplastic samples which bear different mutated alleles.
'ELUSIVE' LITERATURE.
•
A LONGITUDINAL STUDY OF HCV RNA DETECTED BY IN SITU RT-PCR IN LIVER BIOPSY SPECIMENS DURING PROGRESSION FROM CHRONIC HCV LIVER DISEASE TO HEPATOCELLULAR CANCER, HCC. G. Lake-Bakaar, TaiPing Lee, P.Ells and G. Nnovo. Depts of Medicine and Pathology, SUNY HSC Stony Brook and VAMC Northport,/flY.
Integrated HBV ganomes are found in most HCCs developing in patients with chronic HBV infection. By contrast, the nssneimion between HCV and related HCC is unclear. In sire rt-PCR can detect low copy numbers of HCV RNA nacleetide sequences in cells, and should therefore identify clanal expansion of HCV bearing cells in developing HCC. We have evaluated sequential liver biopsies from a patient JM, for the presence and distribution ofHCV RNA using in sire rt-PCR. HISTORY: Patient JM firstprosnated in 1984, aged 60 years with abnormal LFTs detected during a routine medical cheek-up. A past histuty of janndice in 1944 whilst a soldier in Italy was volunteered. HBV serology and other markers of chronic hepatitis were negative. A percumneons liver biopsy was performed and a presumptive diagnosis of non-A, non-B hepatitis made. JM remained asymptomatic for the next several years. In 1990, anti-HCV was detected with first generation antibody tests. In 1992, a CT scan of the abdomen revealed a normal liver and spleen. Physical examination in February 1994 revealed a subcutaneous nodule cephalad m the umbilicus, in the midiine. Serum alpha feto-protein was 195,000 IU/ml. An abdominal ultrasound showed a 6 cm hypnechoic mass in the dome of the right lobe of the liver with portal vein thrombosis as well as cavernous transformation nt the veins at the porta hepatis. Abdominal CT confirmed portal vein thrombosis with collateral filing and revealed a peripancreatic and a celiac lymph node. MRI was consistent with a mniticentric HCC. In March 1994, a percutancous liver biopsy was performed which showed cirrhosis without minor. Subsequently, a CT guided biopsy of the mass was performed which revealed HCC. METHODS: In situ rt-PCR was performed on liver biopsy tissue mounted on glass slides ares digestion with trypsin, followed by overnight treatment with RNAse-free DNAse. The tissues were incubated at 42°C for 30 mins with the downstream primer and reverse transcriptnse. Insim PCR was then performed with a solution containing 4.5 MgCl2, 200uM dNTPs, 10uM digoxiganin dUTP and luM of the primers. RESULTS: The biopsy obtained in 1984 showed moderate hepatocellular necrosis, mild focal large droplet fatty change and portal tract expansion with an increased number of lymphocytes and a striking eosinophilic component. HCV cDNA was present in hepatnsytes within several lobules in a panlobular distribution. The percotaneous liver biopsy from March 1994 showed micronedular cirrhosis without tumor or alphafetoprntein positive cells. HCV eDNA was also present in scattered hepatocytes. By contrast in the CT guided biopsy of the HCC mass which revealed HCC, no HCV cDNA was detected in the turnout tissue. CONCLUSION: These data do not suggest that the transformation from chronic HCV liver disease to HCC is associated with persistence of HCV nucleotide sequences in HCC cells.
AGA ABSTRACTS
GASTROENTEROLOGY, VOl. 108, NO. 4
ENDOSCOPIC, L A P A R O S C O P I C A N D OPEN S U R G E R Y FOR
IN P U R S U I T O F
EARLY GASTRIC CANCER. K. Kumai, Y. Otani, M. Ogami, T, Kubota, M. Kitajima. Depts. of Surgery, School of Medicine, Keio University, Tokyo, Japan
A Kumar, Division of Gastroenterology, Univ of Tennessee at Memphis. Completeness of retrieval of studies is imperative for good oncological epidemiology. MEDLINE search does not retrieve a significant number of articles.Beginners do not have access to not easily available [Elusive] literature.We investigated these sources of information. M~ETHOD:Review of available literature SOURCES OF ELUSIVE L I T E R A T U R E - Published works: subject catalogues, abstracts, indexes,reviews, databases [EXCERPTA, EMBASE]. Retrieval continued till authors names appear again [&again].Completed works:Papers/technical reports-National Technical Information Center [NTIS] database, 26 areas, 88% completion, abstract newsletter weekly: Educational Resources Information Center[ERIC] database :Federal Database Finder [Information,USA, 1986]: Monthly catalog of US Government Publications[Office Superintendent Documents] Dissertations/theses-DATRIX II&CDI [Univ Microfilm International, AnnArbor,MI]: Bibliography Index[Oryx Press 1985]MeetingsBibliographic Guide [G K Hall,Boston], Conference Papers Index[Cambridge Scientific Abstracts,Bethesda]Census/economic dataCensus Information Census:Economic Bulletin Board:Population Bibliography:Statistical Analyses [Elveiser1991]Studies in progress: Federal Reseach In Progress[NTIS]:UNIVRES [Canada],CRIB[Britain] and STRC[Australia] {Elveiser1991}:EUDISED[Europe]:FORIS[Germany] Informal means-meetings,electronic networks, public policy organisations,researchers in same field.
The majority of mucosal gastric cancers could be curatively treated by local resection of the stomach without lymph node dissection, because lymph node metastases were observed in only 10/475 patients(2.0%). As there was no lymph node metastasis in the cases who had lesions less than 2.5 cm in diameter in our department, Endoscopic Mucosal Resection (EMR) has been applied for 70 patients with an elevated mucosal lesion less than 2 cm in diameter or with a depressed lesion less than 1 cm, but 5 cases were converted to open surgery technically. Therefore we limited the indication of EMR for a lesion less than 1 cm in diameter. And as a new method, we t r e a t e d l 8 patients by 2 laparoscopic procedures. Laparoscopic wedge resection of the stomach was performed for 9 patients by Lesion Lifting Method. The gastric lesion was lifted laparoscopically using the string which was pierced the gastric wall just near the lesion. Wedge resection w a s c a r r i e d out with a sufficient surgical margin. Laparoscopic Intragastric Mucosal Resection was also performed for 9 cases who had lesions at the posterior wall or near the cardia or the pyloric ring. Open surgery using modified lymph node dissection could be applied for other mucosal cancers. A standard radical lymph node dissection should be need for submucosally invaded cancers, because 13.5% of lymph node metastasis was observed.
K~RAS2 C O D O N 12 G A T AND G T T C O E X I S T I N G SSCPs OCCUR FREQUENTLY IN PERIAMPULLARY N E O P L A S M S OF P A N C R E A T I C AS W E L L AS O F B I L I A R Y ORIGIN. L.Laghi, L. Arborati, O. Orbetegli, M. Sampiotro, A. Zerbi, D. Faravelli, V.Di Carlo, A. Malesci, G. Fiorelli. Istituto di Medicina Intema e Fisiopatologia Medica, Ospedale Policlinico; Clinica Chirurgica e Anatomia Patologica, Istituto S. Raffaele, Milano, Italy. Pancreatic cancer has the highest prevalence of Kras2 codon 12 mutations of any human nooplasm.Transve:'sion to G A T accounts for 50% of mutations, and transversion to G T T for 30%, while other substituitions are reportod with variable frequency. Coexisfiug mutations are rarely (<5%) described. More heterogeneous data are available for bile duct neoplasms, where coexisting mutations (mainly G A T and GTT) occur to varying degrees (30-100%). AIM: We assessed whether periampullary pancreatic and biliary tumors would produce different Single Straod Conformation Polymorphism (SSCP) patterns of Kras2 exonl, and pm'dcularly patterns homologous to G A T aud G ' I T codonl2 mutations. M E T H O D S : 56 specimens of periampullary neoplasms have been processed. Using a heminestod PCR technique, SSCPs of 107bpamplic0ns (spanniug the nt -13 +93 in the first exon of Kras2) were obteined from paraffin-embedded neoplastic tissues and compared with D N A s control with known G A T and G T T mutations. SSCPs were detected by silver staining of 15% PAGE-minigeI employing conditions standardized to detect both mutations simultaneously. RESULTS: SSCP patterns different from normal were detected in 34141(83%) pancreatic ductal neoplasms,0/2 ampollary neoplasms, 4/4 acinar neoplasms and i0/10 distal bile duct. O b s e r v e d SSCP p a t t e r n s T u m o r s type GAT GTI" IGAT+GTT Other Bilim7 0 0 7(70%) " 3(30%) DuctalPancreatic 9(26%) 9 (26%) 12 (35%) 4 (13%) AmpullatT 0 0 0 0 Acinar 1(25%) 0 3 (75%) 0 Coexistence of different SSCP patterns (mainly GAT and GTT-like) in pancreatic ductal-originating tumors was associated with the presence of mucinous areas (7/9 vs 7125, X2=0.025). C O N C L U S I O N S : Our data suggest dmt 1) multiple coexisting Kras cod 12 mutations occur in bile duct neoplasms ; 2) double mutations in ductal-ofiginating tumors of the pancreas are more frequently seen in mucinous areas; 3) also acinar lesions can progress through these genetic changes. Microdissection studies can clarify if it is possible recognize diffelrent areas in neoplastic samples which bear different mutated alleles.
'ELUSIVE' LITERATURE.
•
A LONGITUDINAL STUDY OF HCV RNA DETECTED BY IN SITU RT-PCR IN LIVER BIOPSY SPECIMENS DURING PROGRESSION FROM CHRONIC HCV LIVER DISEASE TO HEPATOCELLULAR CANCER, HCC. G. Lake-Bakaar, TaiPing Lee, P.Ells and G. Nnovo. Depts of Medicine and Pathology, SUNY HSC Stony Brook and VAMC Northport,/flY.
Integrated HBV ganomes are found in most HCCs developing in patients with chronic HBV infection. By contrast, the nssneimion between HCV and related HCC is unclear. In sire rt-PCR can detect low copy numbers of HCV RNA nacleetide sequences in cells, and should therefore identify clanal expansion of HCV bearing cells in developing HCC. We have evaluated sequential liver biopsies from a patient JM, for the presence and distribution ofHCV RNA using in sire rt-PCR. HISTORY: Patient JM firstprosnated in 1984, aged 60 years with abnormal LFTs detected during a routine medical cheek-up. A past histuty of janndice in 1944 whilst a soldier in Italy was volunteered. HBV serology and other markers of chronic hepatitis were negative. A percumneons liver biopsy was performed and a presumptive diagnosis of non-A, non-B hepatitis made. JM remained asymptomatic for the next several years. In 1990, anti-HCV was detected with first generation antibody tests. In 1992, a CT scan of the abdomen revealed a normal liver and spleen. Physical examination in February 1994 revealed a subcutaneous nodule cephalad m the umbilicus, in the midiine. Serum alpha feto-protein was 195,000 IU/ml. An abdominal ultrasound showed a 6 cm hypnechoic mass in the dome of the right lobe of the liver with portal vein thrombosis as well as cavernous transformation nt the veins at the porta hepatis. Abdominal CT confirmed portal vein thrombosis with collateral filing and revealed a peripancreatic and a celiac lymph node. MRI was consistent with a mniticentric HCC. In March 1994, a percutancous liver biopsy was performed which showed cirrhosis without minor. Subsequently, a CT guided biopsy of the mass was performed which revealed HCC. METHODS: In situ rt-PCR was performed on liver biopsy tissue mounted on glass slides ares digestion with trypsin, followed by overnight treatment with RNAse-free DNAse. The tissues were incubated at 42°C for 30 mins with the downstream primer and reverse transcriptnse. Insim PCR was then performed with a solution containing 4.5 MgCl2, 200uM dNTPs, 10uM digoxiganin dUTP and luM of the primers. RESULTS: The biopsy obtained in 1984 showed moderate hepatocellular necrosis, mild focal large droplet fatty change and portal tract expansion with an increased number of lymphocytes and a striking eosinophilic component. HCV cDNA was present in hepatnsytes within several lobules in a panlobular distribution. The percotaneous liver biopsy from March 1994 showed micronedular cirrhosis without tumor or alphafetoprntein positive cells. HCV eDNA was also present in scattered hepatocytes. By contrast in the CT guided biopsy of the HCC mass which revealed HCC, no HCV cDNA was detected in the turnout tissue. CONCLUSION: These data do not suggest that the transformation from chronic HCV liver disease to HCC is associated with persistence of HCV nucleotide sequences in HCC cells.