Nobuo Yamada

Activities of free oxygen radical scavenger enzymes in human liver

Activities of superoxide dismutase, catalase and glutathione peroxidase were measured in liver biopsy specimens from patients with various liver diseases, including six with chronic persistent hepatitis, nine with chronic active hepatitis, nine with non-alcoholic cirrhosis, eight with alcoholic cirrhosis and eight with acute hepatitis. Measurements from ten patients without liver disease were used as controls. Levels of total superoxide dismutase activity in the chronic active hepatitis and non-alcoholic cirrhosis groups were significantly lower than those in the controls (p less than 0.01 and p less than 0.01, respectively). The level of total superoxide dismutase activity in the acute hepatitis group was significantly higher than that in the control group (p less than 0.01). The levels of Cu,Zn-superoxide dismutase activity in all the experimental groups, except for the chronic persistent hepatitis group, were significantly lower than those in the controls (p less than 0.01 in all groups). The levels of Mn-superoxide dismutase activity in the alcoholic cirrhosis and acute hepatitis groups were significantly higher than those in the controls (p less than 0.01 and p less than 0.01, respectively), although no difference in the level of this enzyme was seen among the controls, chronic persistent hepatitis, chronic active hepatitis and non-alcoholic cirrhosis groups. The levels of catalase activity in the groups with chronic active hepatitis, non-alcoholic and alcoholic cirrhosis and acute hepatitis were significantly lower than those in the controls (p less than 0.01 in all groups). Glutathione peroxidase activity showed no difference among the groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Treatment of symptomatic hepatic cysts by percutaneous instillation of minocycline hydrochloride

Nine patients with 16 symptomatic nonneoplastic congenital hepatic cysts were treated prospectively by ultrasonically guided percutaneous minocycline hydrochloride injection, and the usefulness of this treatment was evaluated. Seven of the patients had multiple hepatic cysts, and two solitary cysts. All the patients were women, ranging in age from 36 to 81 years. After cystic fluid had been aspirated with a 21-gauge PTC needle, minocycline hydrochloride was injected into all the cysts. The minocycline hydrochloride was dissolved in saline at a concentration of 200 mg in 9 ml, and mixed with 1 ml of 2% mepivacaine hydrochloride. The total quantity of minocycline hydrochloride injected varied from 100 mg to 1200 mg per hepatic cyst, depending on its size. Total or subtotal regression of the cysts was observed in all patients during follow-up periods ranging from 15 to 35 months. Seven patients became symptom-free, one showed symptom reduction, and one showed no change in symptoms. Minor side effects, eg, transient abdominal pain, slight right shoulder pain, and temperature elevation, were noted in three patients respectively. On the basis of these results, we conclude that ultrasonically guided percutaneous minocycline hydrochloride injection is useful for the treatment of symptomatic hepatic cysts.

Detection of Hepatitis G virus RNA in patients with Hepatitis B, Hepatitis C, and non-A-E Hepatitis by RT-PCR using multiple primer sets

Hepatitis G virus(HGV)/GB virus C(GBV‐C) is a newly identified virus associated with human hepatitis. The preliminary prevalence studies of HGV infection in Japan were entirely based on the detection of HGV RNA by RT‐PCR. However, the selection of the different primer sets in such assay may influence sensitivity of the test because of the extensive genetic heterogeneity of HGV, and influence the estimation of the prevalence of HGV. To address this potential problem, we designed two primer sets from well conserved domains in the 5′NC and NS5 regions of HGV genome, and tested them together with the NS3‐derived primer set in RT‐PCR for their ability to detect HGV RNA in serial dilution of synthetic viral RNA templates. Subsequently, we used these three primer sets to detect HGV RNA in the sera of 371 Japanese patients with hepatitis B, hepatitis C, and non‐A‐E hepatitis. The results indicated that the primer set derived from the 5′NC region appeared to be most effective in detecting HGV RNA. The results also showed that only two out of the 126 patients (1.6%) with non‐A‐E hepatitis were positive for HGV RNA although the RNA were detected more frequently in patients with hepatitis B (2/38; 5.3%) and hepatitis C (17/207; 8.2%), suggesting that HGV is not a common causative agent for non‐A‐E hepatitis in Japan. J. Med. Virol. 52:385–390, 1997.

Spatiotemporal Measurement of Free Radical Elimination in the Abdomen Using an In Vivo ESR-CT Imaging System

Electron spin resonance (ESR) imaging can visualize the distribution of free radicals in living systems according to their concentrations. However, the application of ESR imaging to living animals has not been well established. Using a rapid field scan L-band ESR imaging system, we have successfully obtained two-dimensional ESR projection (xz-plane projection) and three-dimensional ESR-CT (trans-axial section along the y-axis) images of the abdomen of living mice after an injection of 3-carbamoyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (carbamoyl-PROXYL) into the tail vein. The in vivo two-dimensional ESR projection imaging clearly visualized the carbamoyl-PROXYL distribution and the rapid decay process in the abdomen. Because among the viscera, the liver is most abundantly associated with a blood volume, the outline of the image can be composed mainly of this organ. We therefore attempted to find whether there will be a difference in spatiotemporal dynamics of carbamoyl-PROXYL in the abdomens between the control and the mice with liver damage by two-dimensional ESR projection. In the control mice, carbamoyl-PROXYL was almost completely eliminated from the abdomen within 5 minutes after administration. On the other hand, in mice with carbon tetrachloride-damaged livers, the decay of carbamoyl-PROXYL was markedly prolonged. Even at 5 min after administration, carbamoyl-PROXYL remained clearly visible in the abdomen. In vivo three-dimensional ESR-CT imaging showed an even distribution of carbamoyl-PROXYL throughout the whole liver, which corresponded well with the images of trans-axial sections of the murine abdomen. We have succeeded in displaying two-dimensional ESR projection and three-dimensional ESR-CT images of carbamoyl-PROXYL distribution and clearance in the abdomen of a living animal. The ESR-CT imaging technique is considered to be a powerful new tool for noninvasive investigations of the in vivo spatiotemporal dynamics of free radical distribution and elimination in the organs.

Case Report: Interferon induced coma in Sheehan's syndrome

A 54‐year‐old woman who was being treated with 10 million units (mu) of natural interferon (IFN)‐α per day for chronic active hepatitis C at a local clinic, developed coma on the fourth day of treatment. On admission to Yamagata University Hospital, she was still in a state of semicoma with severe hyponatraemia (122 mEq/L) and hypochloraemia (89 mEq/L). After the administration of electrolytes, her condition improved remarkably. Endocrinological loading tests showed a hypofunction of the anterior pituitary gland. In consideration of these results, and her past experiences of haemorrhage during childbirth and subsequent amenorrhoea, we diagnosed her illness as a coma as a result of Sheehans syndrome which had become overt during IFN therapy. She recovered completely after treatment with hydrocortisone and 1‐thyroxine.

Computerized morphometry of liver cells in diffuse hepatic diseases

SummaryComputerized morphometrical measurements were made of liver cells and their nuclei taken from livers of healthy persons and from patients with acute hepatitis (AH), chronic hepatitis (CH), and liver cirrhosis (LC). Liver biopsies were performed on 127 people (24 convalescing from acute hepatitis, 20 with chronic inactive hepatitis (CIH), 24 with chronic active hepatitis (CAH), 38 with liver cirrhosis (LC) and 21 who were healthy controls (HC)). The specimens were stained with hematoxylin-eosin, then light microscopic pictures were obtained.The original cells were magnified 1000 times. The sizes of the cells and of their nuclei were calculated using a computer. Correlative variation was noted in cell and nuclear sizes resulting in an consistently constant nucleus-cytoplasm ratio. Average sizes of both cells and nuclei were maximal in livers with LC, followed in decreasing order by those in livers with CAH, CIH, HC and convalescent AH. The smallest cells and nuclei were found in convalescent AH cases. It might be possible to some degree to obtain gross impressions of the histopathological type of the liver lesion by measuring the sizes of cells and nuclei of the liver.

Quantitative Measurement of Fluorescent Intensity of the Rat Liver Surface Using Video Fluorescence Laparoscopy Following Intravenous Fluorescein Injection

Abstract: A new method for the quantitative measurement of fluorescent intensity of the rats liver surface after fluorescein injection is described. The measuring system of fluorescent intensity on the rat liver surface consisted of a video laparoscopy with a video densitometer. An excitation filter was placed in the light source and an absorbing filter was attached to the eyepiece of the magnifying scope in this fluorescence video laparoscopy system. Fluorescein was injected intravenously into the inferior vena cava. Video fluorescence images of the liver surface were recorded with a CCD TV camera which was connected to the scope. In measuring the mean intensity of video images, a region of interest was designated on the TV monitor. The fluorescent intensity in this region was measured by a video densitometer and expressed in volts. The time‐intensity curve was graphically presented using a pen recorder. In this way, the time course of the liver surface fluorescent intensity could be analyzed with data expressed in volts. In normal livers, the fluorescent intensity began to rise 4.0 ± 0.5 seconds after fluorescein injection and reached a peak at 14.4 ± 1.5 seconds. Thereafter, the intensity fell slightly but then rose again to a second peak 170±50 seconds after injection.