- Email: [email protected]
Automated histological evaluation of gastric biopsy specimens using digital slide
described the are't, cell density', cellular characteristics of the basic tissue components: the surface epithelium, the glands, the muscle and connective tissue and the inflammatory cell compartment in the biopsy. In each compartment the cell morphometric features were calculated, as well. Area ratios of the dillS:rent tissue compartments (biopsy area to epithelia, gland, connective tissue area, epithelium to connective tissue, epithelium to muscle tissue, epithelium to glands, epithelium-and glands to non epithelial tissue) were also calculated. Linear discriminant analysis was used for classification of the samples (Statistics, V.6.06, Sta~oft, USA) into the diagnostic criteria ( normal, gastritis, adenocarcinoma). Results: Siginlicatu differences were found between the ratios of the biopsy/gLand area ( 4,68 + 1.65 in normal, 4.04+ 1,05 in gastritis, and 67.5 +48.5 in adenocarcinotna (p <0.05)), and in the biopsy/connective tissue area (4.4 + 3.9, 1.73 + 0.37, 2.75 + 1.32, p 0.05). Significant difference were also tound in the ratio of cell numbers in the different cell compartments, too( 5.65+2.65 in heahhy,16,7+7.4 gastritis, 180.1,+114.2 adenocarcinoma p
of power Doppler US examination by comparison with endoscopic disease activity in patients with UC. (Methods) A total ot 129 patients (mean age 38.4 (range 17-72), 51 male, 78 female) with UC were studied The diagnosis of UC was established with standard clinical, radiologic, histologic, and endoscopic criteria (Matts grade). Power Doppler US was perforned by using 60-11.0 MHz transducer, the lowest possible pulse repetition frequency without aliasing, a low wall filter, and high Doppler gain settings. Power Doppler pixels were counted on the view of the most thickened wall. Pulsed Doppler US was used to &stingnish the blood flow siguals from wall motion artiLacts. Colonoscopy was also done within one week alter US stud?/"to evaluate bowel acti~4ty The relationship between power Doppler signals and endoscopic criteria of most thickened wall were assessed. (Result) Average count of power Doppler pixels in Marts grade 1 (53 patients) was 40.45, Marts grade 2 (36 patients) was 66.69, Marts grade 3 (37 patient) was 1186, Marts grade 4 (3 pauems) was 1365. As shown in Table, patients with higher endoscopic activity (Matts grade 3 and 4) had significantly more pixel counts than those of patients with lower activity (Marts grade 1 and 2). (p<0.0001) (Conclnsion) Power Doppkr pixels of most thickened wall were closely related to endoscopic disease acti~Aty in patients with UC.
P
I Pixeleotmt
]
P=O.0(O6
I
M2164 Clinical Significance of Porta Hepatis Lymphadenopathydetected by Endoscopic Ultrasound (EUS)
P=0.4942
I
[--1
Alexis Ayonote, Rosario Ligresti
4000
BACKGROUND AND OBJECTIVE: Porta hepatis lymphadenopathy is frequendy seen on EU& We undertook this analysis to evaluate the clinical significance of porta hepatis lymphadenopathy visualized at the time of EUS for other indications, MATERIALS, PATIENTS AND METHODS: The study period was from April 1999 to February 2002. A retrospectwe analy'sis was done of 530 patients who had EUS pertbrmed for other indications at the West&ester Medical Center and Jacobi Medical Center by a single experienced endnscopist/ endosonograpber. The data was analyzed by Student t-test, chi-square, and multiple linear regression. After informed consent, EUS was pertbrraed with a radial scanning echoendoscope (GFUM 20, Olympus America Inc., Melville NY) at 7 5 MHz. Porta hepatis lymphadenopathy was assessed for by scanning through the duodenal bulb, RESULTS: EUS was performed on 530 patients, 64 patients (12%) had porta hepatis lyraphadenopathy. 38 patients (59.4%) were female, 26 patients (40.6%) were male The mean age of patients was 55.Syears (2885years).The mean size of porta hepatis lymph nodes(LN)16.6mm(6-59mm). 40.6% were isoecboic and 59,4% were reactive appearing lymph nodes. The mean common bile duct (CBD) size 62mm(2.8-22mm). Of the 64 total patients-7 had abdominal pain(11%,mean LN size 18.8mm),12 had panereatitis(19%,mean LN size 16,4mm), 3 had hepatitis C(HCV) (5%, mean LN size 26.7mm), 1 had hepatitis B(HBV)(2%, mean LN size 12ram), 9 had either bfliary or pancreatic cancers,(14%, mean LN size 11.8mm), 3 had HlV(5%,mean LN size 13.7mm), 30 had CBD and/or gall bladder stone disease(47%,mean LN size 17ram). Mean LN size was higher in patients with HCV(26,7mm)(p=O,06).Indicatinn for the test did not correlate with LN size(p = 0.52), CBD size(p = 0.06) or LN description(p = 0.79). LN size did not correlate with LN description(p = 0.29). CBD size did not correlate with LN description(p = 0.80), but did correlate with LN size(p<0.O01). Mean size of LNs was higher in the isoeehoic group vs reactive group(18.4mm vs 15.3mm, p<0.O01). CONCLUSIONS: In summary, porta hepatis lymphadenopatby is found more often in patients with gallstone biliary disease, pancreatitis,and biliary or pancreatic cancers. Larger porta hepatis lympadenopathy is more common in patients with dilated common bile duct, gaUstone bfliary disease, or hepatitis C These lymph nodes tended to be more isoeehoic, but overall, EUS lymph node description could not be used as an independent predictor of associated pathology'.
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3500 3000
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1000 500
0
1
2
3
4
Mat~ grade
M2162
New, Rapid Fluorescence Stain Method for Histologic Sections Using Lanthanide Complexes J.-G. Wu, Y.-L Su, Roger D. Soloway, J. Yang, Y. Ran, Y:Z. Xu, J. Wang, J.-S. Wang, L. Zhang, Z, Liu, L.-M, Yang, F, Wang, W,-H. Li, D,-F. Xu, Q.-G. Wu We have used Europium (Eu, MW 152, element #63) and Terbium (Th, MW 156, element #65), from the lanthanide series, to form complexes with excellent fluorescence, One group of complexes was tormed with the antibiotic ofloxacin (O). These complexes are watersoluble and emit a bright green or a blue iluorescence under UV (1>320 nm) light, Traditional staining with Hematoxyhn and Eosin (H and E) may require as many as ten steps, This method speeds up the process and adds new spectral information. The aim of this study was to examine the use of newly synthesized lanthanide complexes, EuO and TbO for tissue staining and spectroscopy. Fixed or frozen specimens were sectioned and stanied with water or chloroform solubfliaed complex for 1-2 minutes followed by washing in either water or chloroform to remove excess dye and then air-dried A second dide was processed by staining with H and E. Fourier translorm infrared (FT-IR) spectra were obtained on a mi& IR spectrometer at 360-x magnification, Results showed a dense collection of lyanphocytes within the liver was stained with ThO; the nuclei were bright green. A liver lobule was stained with ToO; nuclei were a light green, cytoplasm a dark green and sinusoids unstained. The same fiver stained with EuO demonstrated dark blue nuclei, light blue cytoplasm, and no staining in sinnsoids. Fluorescence spectra demonstrated sharp peaks for the complex at 490 and 545 cm ~ and smaller peaks at 583 and 620 cm Neither the peak position nor bandwidth at peak halgheight was significantly changed by binding to tissue, However, the reLative peak height at 550 was decreased by binding to fiver tissue and this demonstrated different ratios of relative intensities (I,va~,Jl~5,~) after the complex bound with tissues: TbO alone 0517, with gallbladder 0.673, with lymphocytes 0.688, with hepatoc)r 0.732 and with splenic tissue 0,766. We conclude that EuO and TbO complexes form a variety of fluorescent binding with tissues and that FT-IR spectroscopy of this tissue produces a variety of changes in relatwe intensity after binding that tray provide a new basis for histologic examination of tissue pathology. "Supported by National Key Project China No. 2002CCA01900"
M2165
Twenty-four-hourEnergy Metabolism in Cirrhosis Kong Chen, Sari Acra, Joseph Awad, Raymond Burk Purpose; Components of 24-hr energy expenditure in cirrhotic patients have not previously been assessed using a metabolic chamber. The purpose of this pilot study was to evaluate whether energy metabolism was altered m adult patients with cirrhosis compared to healthy, controls. Methods: Subjects: 7 adult patients, ages 42-57, with established, compensated histologieally-proven cirrhosis (CIR) from difterent causes (hepatitis C, alcohol, and autolmmune), but free of ascites, and encephalopathy; 7 age, gender, weight and body compositionmatched healthy controls (H). Protocol: Components of energy expenditure (basal, physical actwity, post-physical actwity and diet-induced energy expenditure) were measured during a 24-hr stay" in a state-of-the-art metabolic chamber fitted with a force platform. Body composition (hi mass, fat-free mass, [FFMI) was assessed using air-displacement plethysmograpby (godPod). Resuhs: Total energy expenditure was significantly lower in ClR vs. H (43.5-+ 5.2 vs. 51,0_+ 3.1 kcal/day/kg FFM, or 2851 _+459 and 3174_+ 560 kcal/day, respectively', [P = 0.021]). Basal (sleeping) energy expenditure was elevated in 5/7 CIR (CIR = 33.3 +- 1.3 vs. H = 29.4_+2.0 kcal/day/kg FFM, [P=0.0351). Thermal effect of food was comparable (CIR = 3,2 +- 1,9 vs. H = 3.3 _+2.8 kcal/day/kg FFM, [P = 0,81). Energy' expenditure by physical activity/24 hrs was significantly lower (P
M2163
Automated Histological Evaluation of Gastric Biopsy Specimens Using Digital Slide kevente Ficsor, Bela Molnar, Peter Gvmlbas, Zsoh Tulassay Background: Digital slides and virtual microscopy ,developed in our laboratory, was shown to be ahematwe techniques for routine gastric biopsy' specimen analysis. Image analysis techniques should support the automated, quantitanve, reproducible classification of digital slides into major histological diagnostic groups. Materials and methods: From routine gastric biopsy', histological material 3 heahhy, 17 gastritis (mild, moderata, severe) and 7 adenoearcinoma , H/E stained rminne sections were selected. Using the Hi4cope (3DHistech Ltd, Budapest, HungaI3') slide digLtiser system, digitalization was peffbrme& In C + + (BorLand, USA) automatic histological evaluation modules were developed, Altogether 36 parameters
AGA
Abstracts
A-428
of power Doppler US examination by comparison with endoscopic disease activity in patients with UC. (Methods) A total ot 129 patients (mean age 38.4 (range 17-72), 51 male, 78 female) with UC were studied The diagnosis of UC was established with standard clinical, radiologic, histologic, and endoscopic criteria (Matts grade). Power Doppler US was perforned by using 60-11.0 MHz transducer, the lowest possible pulse repetition frequency without aliasing, a low wall filter, and high Doppler gain settings. Power Doppler pixels were counted on the view of the most thickened wall. Pulsed Doppler US was used to &stingnish the blood flow siguals from wall motion artiLacts. Colonoscopy was also done within one week alter US stud?/"to evaluate bowel acti~4ty The relationship between power Doppler signals and endoscopic criteria of most thickened wall were assessed. (Result) Average count of power Doppler pixels in Marts grade 1 (53 patients) was 40.45, Marts grade 2 (36 patients) was 66.69, Marts grade 3 (37 patient) was 1186, Marts grade 4 (3 pauems) was 1365. As shown in Table, patients with higher endoscopic activity (Matts grade 3 and 4) had significantly more pixel counts than those of patients with lower activity (Marts grade 1 and 2). (p<0.0001) (Conclnsion) Power Doppkr pixels of most thickened wall were closely related to endoscopic disease acti~Aty in patients with UC.
P
I Pixeleotmt
]
P=O.0(O6
I
M2164 Clinical Significance of Porta Hepatis Lymphadenopathydetected by Endoscopic Ultrasound (EUS)
P=0.4942
I
[--1
Alexis Ayonote, Rosario Ligresti
4000
BACKGROUND AND OBJECTIVE: Porta hepatis lymphadenopathy is frequendy seen on EU& We undertook this analysis to evaluate the clinical significance of porta hepatis lymphadenopathy visualized at the time of EUS for other indications, MATERIALS, PATIENTS AND METHODS: The study period was from April 1999 to February 2002. A retrospectwe analy'sis was done of 530 patients who had EUS pertbrmed for other indications at the West&ester Medical Center and Jacobi Medical Center by a single experienced endnscopist/ endosonograpber. The data was analyzed by Student t-test, chi-square, and multiple linear regression. After informed consent, EUS was pertbrraed with a radial scanning echoendoscope (GFUM 20, Olympus America Inc., Melville NY) at 7 5 MHz. Porta hepatis lymphadenopathy was assessed for by scanning through the duodenal bulb, RESULTS: EUS was performed on 530 patients, 64 patients (12%) had porta hepatis lyraphadenopathy. 38 patients (59.4%) were female, 26 patients (40.6%) were male The mean age of patients was 55.Syears (2885years).The mean size of porta hepatis lymph nodes(LN)16.6mm(6-59mm). 40.6% were isoecboic and 59,4% were reactive appearing lymph nodes. The mean common bile duct (CBD) size 62mm(2.8-22mm). Of the 64 total patients-7 had abdominal pain(11%,mean LN size 18.8mm),12 had panereatitis(19%,mean LN size 16,4mm), 3 had hepatitis C(HCV) (5%, mean LN size 26.7mm), 1 had hepatitis B(HBV)(2%, mean LN size 12ram), 9 had either bfliary or pancreatic cancers,(14%, mean LN size 11.8mm), 3 had HlV(5%,mean LN size 13.7mm), 30 had CBD and/or gall bladder stone disease(47%,mean LN size 17ram). Mean LN size was higher in patients with HCV(26,7mm)(p=O,06).Indicatinn for the test did not correlate with LN size(p = 0.52), CBD size(p = 0.06) or LN description(p = 0.79). LN size did not correlate with LN description(p = 0.29). CBD size did not correlate with LN description(p = 0.80), but did correlate with LN size(p<0.O01). Mean size of LNs was higher in the isoeehoic group vs reactive group(18.4mm vs 15.3mm, p<0.O01). CONCLUSIONS: In summary, porta hepatis lymphadenopatby is found more often in patients with gallstone biliary disease, pancreatitis,and biliary or pancreatic cancers. Larger porta hepatis lympadenopathy is more common in patients with dilated common bile duct, gaUstone bfliary disease, or hepatitis C These lymph nodes tended to be more isoeehoic, but overall, EUS lymph node description could not be used as an independent predictor of associated pathology'.
0
3500 3000
2500 2000 ] 500
m
1000 500
0
1
2
3
4
Mat~ grade
M2162
New, Rapid Fluorescence Stain Method for Histologic Sections Using Lanthanide Complexes J.-G. Wu, Y.-L Su, Roger D. Soloway, J. Yang, Y. Ran, Y:Z. Xu, J. Wang, J.-S. Wang, L. Zhang, Z, Liu, L.-M, Yang, F, Wang, W,-H. Li, D,-F. Xu, Q.-G. Wu We have used Europium (Eu, MW 152, element #63) and Terbium (Th, MW 156, element #65), from the lanthanide series, to form complexes with excellent fluorescence, One group of complexes was tormed with the antibiotic ofloxacin (O). These complexes are watersoluble and emit a bright green or a blue iluorescence under UV (1>320 nm) light, Traditional staining with Hematoxyhn and Eosin (H and E) may require as many as ten steps, This method speeds up the process and adds new spectral information. The aim of this study was to examine the use of newly synthesized lanthanide complexes, EuO and TbO for tissue staining and spectroscopy. Fixed or frozen specimens were sectioned and stanied with water or chloroform solubfliaed complex for 1-2 minutes followed by washing in either water or chloroform to remove excess dye and then air-dried A second dide was processed by staining with H and E. Fourier translorm infrared (FT-IR) spectra were obtained on a mi& IR spectrometer at 360-x magnification, Results showed a dense collection of lyanphocytes within the liver was stained with ThO; the nuclei were bright green. A liver lobule was stained with ToO; nuclei were a light green, cytoplasm a dark green and sinusoids unstained. The same fiver stained with EuO demonstrated dark blue nuclei, light blue cytoplasm, and no staining in sinnsoids. Fluorescence spectra demonstrated sharp peaks for the complex at 490 and 545 cm ~ and smaller peaks at 583 and 620 cm Neither the peak position nor bandwidth at peak halgheight was significantly changed by binding to tissue, However, the reLative peak height at 550 was decreased by binding to fiver tissue and this demonstrated different ratios of relative intensities (I,va~,Jl~5,~) after the complex bound with tissues: TbO alone 0517, with gallbladder 0.673, with lymphocytes 0.688, with hepatoc)r 0.732 and with splenic tissue 0,766. We conclude that EuO and TbO complexes form a variety of fluorescent binding with tissues and that FT-IR spectroscopy of this tissue produces a variety of changes in relatwe intensity after binding that tray provide a new basis for histologic examination of tissue pathology. "Supported by National Key Project China No. 2002CCA01900"
M2165
Twenty-four-hourEnergy Metabolism in Cirrhosis Kong Chen, Sari Acra, Joseph Awad, Raymond Burk Purpose; Components of 24-hr energy expenditure in cirrhotic patients have not previously been assessed using a metabolic chamber. The purpose of this pilot study was to evaluate whether energy metabolism was altered m adult patients with cirrhosis compared to healthy, controls. Methods: Subjects: 7 adult patients, ages 42-57, with established, compensated histologieally-proven cirrhosis (CIR) from difterent causes (hepatitis C, alcohol, and autolmmune), but free of ascites, and encephalopathy; 7 age, gender, weight and body compositionmatched healthy controls (H). Protocol: Components of energy expenditure (basal, physical actwity, post-physical actwity and diet-induced energy expenditure) were measured during a 24-hr stay" in a state-of-the-art metabolic chamber fitted with a force platform. Body composition (hi mass, fat-free mass, [FFMI) was assessed using air-displacement plethysmograpby (godPod). Resuhs: Total energy expenditure was significantly lower in ClR vs. H (43.5-+ 5.2 vs. 51,0_+ 3.1 kcal/day/kg FFM, or 2851 _+459 and 3174_+ 560 kcal/day, respectively', [P = 0.021]). Basal (sleeping) energy expenditure was elevated in 5/7 CIR (CIR = 33.3 +- 1.3 vs. H = 29.4_+2.0 kcal/day/kg FFM, [P=0.0351). Thermal effect of food was comparable (CIR = 3,2 +- 1,9 vs. H = 3.3 _+2.8 kcal/day/kg FFM, [P = 0,81). Energy' expenditure by physical activity/24 hrs was significantly lower (P
M2163
Automated Histological Evaluation of Gastric Biopsy Specimens Using Digital Slide kevente Ficsor, Bela Molnar, Peter Gvmlbas, Zsoh Tulassay Background: Digital slides and virtual microscopy ,developed in our laboratory, was shown to be ahematwe techniques for routine gastric biopsy' specimen analysis. Image analysis techniques should support the automated, quantitanve, reproducible classification of digital slides into major histological diagnostic groups. Materials and methods: From routine gastric biopsy', histological material 3 heahhy, 17 gastritis (mild, moderata, severe) and 7 adenoearcinoma , H/E stained rminne sections were selected. Using the Hi4cope (3DHistech Ltd, Budapest, HungaI3') slide digLtiser system, digitalization was peffbrme& In C + + (BorLand, USA) automatic histological evaluation modules were developed, Altogether 36 parameters
AGA
Abstracts
A-428