Subcellular localization of hepatitis B viral core- and X-protein during the cell cycle in human...
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492A AASLD ABSTRACTS HEPATOLOGY October 1995
1541 DAILY MODIFICATIONS OF SPLANCHNIC HEMODYNAMICS (SH) IN CIRRHOTICS WITH DIFFERENT DEGREE OF VARICEAL BLEEDING (VB) RISK. S.Sirineo. F.Piseaglia. S.Zironi. S.Gaiani. L.Gramantieri. S.Sofia. N,Venl~rolL A.Casal~. R.Cerinaldcsi. L.Bolundi. Isfituto di Clinics Medics e Gastroeatteroiogia, Univ. Bologna, Italy.
We evaluated the pattern of VB in 181 consecutive admissions: VB occurred with a biphasie diurnal pattern, with two peaks at 9:45 &m. and 9:45 p.m. (Cosiocr test p=0.02). To assess possible correlations between changes in SH and VB, we assessed the 24 hrs modifications of portal blood flow velocity (PV) and superior mesenteric artery (SMA RD, by duplex-Doppler ultrasound, performed every 4 hrs, in 16 patients with cirrhosis. The pts were grouped according to the endoscopic VII risk (7 with F0-FI entices=low risk, 5 with F2- F3 varic¢~-.-kigh risk) and to the presence of significant porto-systemic .shunting (SS= 4 pts), mainly the paranmbilical vein, RESULTS: F0-F1 pts showed significant diurnal changes (table 1, p<0.001) of PV and SMA RI (in opposite direction), both above and below baseline OD= 8:00 hr), showing a still partially compliant portal systera. Also F2-F3 pts had significant changes (table 2, p<0.001) of PV and SMA RL However PV only decreased with respect to baseline (lack of portal system compliance), being highest at 8:00 a.n~ and g:o0 p.m., the times that precede the peaks of VB incidence. Moreovex in these latter pts the SMA RI reductions don't cun'espond to PV increases, as observed in the other two groups. In SS pts only PV significunfly changed (p<0.001) above and below baseline.
g ,,r I
12 16 20 24 4 $ 12 16 20 24 4 8
CONCLUSIONS. Pts with F2-F3 vmicos, who have a high VB risk, have different daily modifications of SH respect to F0-F1 and SS pts. In F2-F3 pts the lack of a PV increase in relation to the SMA RI decreases could imply a diversion of blood flow into gnstroesophageal collaterals. Morevoer the F'2-F3 pts diurnal Immndyanmic pattern may partially explain the biphasic occurca~ of VB.
1542 SUBCELLULAR LOCALIZATION OF HEPATITIS B VIRAL CORE- AND X -PROTEIN DURING THE CELL CYLE IN HUMAN HEPATOMA Huh7 CELLS. H Sirma, O Rosmorduc, D Kremsdorf and C Br6chot. INSERM U370, CHU Necker, Paris.
The subcellular distribution of C- and X-proteins has led to conflicting data: exclusively nuclear and cytoplasmic or bilocal. A recent study showed a cell cycle regulated nuclear translocation of C-protein. Major problems in the interpretation of these studies were I) use of non- hepatocytic cell lines; 2) low expression of C and X. We have recently shown accumulation of C and X in stably transfected Huh7 cells with a defective HBV (dHBV) genome, generated from singly spliced HEV RNA (Hepatology, in press). In view of functional implications of C- and X-proteins sublocalization, we restudied their cellular distribution during the cell cycle in these cells. Methods: 1) Huh7 cells were stably transfected with HBV- and/or dHBV-DNAs; 2) Expression of C- and X-Protein was studied by immunoprecipitation and western blot; 3) Cell synchronization was achieved by TGFSl (G1), double Thymidine block [dT] (S) and Nocodazol [N] (M). S-Phase was additionally labelled by BrdU pulse. In situ subcellular localization of C- and X-Proteins was studied by indirect immunofluorescence and konfocal analysis. Results: The clones could be synchronized to higli degree in GI-, S- and M-Phase of the cell Cycle. X-Protein was distributed in asynchronous cells with a perinuclear dominated corona and sometimes as granules embedded in diffuse cytoplasmic staining. TGFB1 and N treatment resulted in a reduced and diffuse cytoplasmic staining pattern. C-Protein was localized perinuclear as a semiclosed ring of homogeneous mass. dT block did not change the characteristic perinuclear sublocalization. Subcellular localization of C- and X- proteins was further confirmed by confocal analysis. Condusions: HBV C- and X-Proteins are sublocated exclusively in the cytoplasm of human hepatoma Huh7 ceils. There is no cell cycle dependent translocation into the nucleus. The mechanism of viral particles assembly and transactivation of cellular genes should be massessed with this information.
1543 EFFECT OF ABDOMINAL PARACENTESIS O N DIAPHRAGMATIC CONTRACTILITY. OJ Smith. SOuiason. G Roisz and W Clmkstun. Die. of Gastroenterology, University of Missouri-KC, Kansas City, MO
Background: Large volume paracentesis (LVP) has long been utilized to relieve dyspnea in patients with tense aseites. The benefit has been suggested to result fi~om improved diaphragnmtic excursion or compliance, but the effect of ascites on diaphragmatic function has undergone limited evaluation. Aim: To determine the effect of abdominal paraoentesis on diaphragmatic contractility. Methods: Ten patients (9 M, 1 F, mean age 43 yr) with alcoholic cirrhosis and tense ascites underwent LVP to relieve dyspnea. Before and after paracentesis the transdiaphragnmtic pressure (Pat), a measurement ofwurk performed by the diaphragm, was determined. Intra- abdominal pressure (P,0 was measured via a gastric balloon catheter attached to a pressure transducer, and intrapleural pressure ( Pp3 was reenrded via an esophageal balloon catheter. Transdiaphragmatic pressure was determined as follows: Pat = P,b - Ppt Results: Dyspnea was relieved in all ten patients, requiring the removal of a mean of 2.5 liters of ascitic fluid. (range 1-5.5 liters). Diaphragmatic contractility was unchanged in one patient, increased in two patients by 3 and 4 mm Hg respectively, and declined in seven patients. Overall transdiaphragmatic pressure (P,0 before paracentesis (mean 37.5 mm Hg) actually declined (mean 35.0 mmHg) as a result of removing ascitic fluid, but the difference was not statistically si~fifioant. (p=0.71).
No. Pts P~ Initial P~i Post A p~ Paracentesis
10 37.5 mmHg 35.0 mmHg -2.0 mmHg
Conclusions: Abdominal paraeentesis has no significant effect on diaphragmatic contractility, as assessed by transdiaphragmatic pressure. The amelioration of dyspnea in patients with tense ascites occurs via non- diaphragmatic mechnnlgm~.
1544 DOES EXCESSIVE ALCOHOL CONSUMPTION WORSEN HCV-LIVER DISEASE'? C Soldevila-Pico. S Haoue. F Rodriguez. R Chanarala. D Gallant. l Penn. ASF Lok. Tulane Unlv and VA Med Ctr, New Orleans, LA.
Retrospective studies suggest that HCV infection may accelerate alcohol Induced liver damage but the effect of excessive alcohol consumption on HCV-liver disease has not been examined. ~ : To determine if excessive alcohol consumption (>80g/d x >5yr) increases the activity/progression of HCV induced liver disease. Patients & Methods : In an ongoing study, liver biopsies were requested on all patients who were found to be anti-HCV+ during their admission for alcohol or drug detoxification. 27 biopsies were reviewed by 2 pathologists who had no knowledge of the clinical/biochemical data. Histological activity and fibrosis were scored according to Knodeli. Results : Of the biochemical and histological markers evaluated, only AST level was significantly different between the patients with alcohol consumption > or < 80g/d.
ETCH consumption < 80 g/d > 80 g/d
No. of patients 12 15 Age (yr) 41.2+1.6 41.9+1.4 ETOH intake (g/d) 39+1 312±77 AST (U/l) (15-37) 39±4 67±10 ALT (U/l) (30-65) 80±10 106±14 GGT (U/I) (5-85) 116±34 238+46 Histological activity score 3.5±0.5 3.1±0.4 Fibrosis score 1.5+0.3 1.5±0.4 Bile duct damage 6796 5396 Steatosis 3396 4796 Perivenular fibrosis 17% 53% Mallory's hyaline 896 2096
Conclusions : In this preliminary analysis, excessive alcohol consumption (>80g/d) did not seem to worsen HCV-liver disease in patients who were asymptomatic for liver disease. Further study is ongoIng to confirm these findings.