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Summary A genetic control of alkaline phosphatase (AP) in chicken duodenum was studied in a White Plymouth Rock strain. Unpurified chicken duodenum AP heated in an extraction procedure comprised either F or S band by isozyme types. On the other hand, chromatographically purified intestinal AP (NBCo, USA) had three bands, i.e., F', S' and B' bands. Characterization by urea, heat and neuraminidase treatments suggested that the genetic control of plasma AP isozymes may be applicable to the duodenum AP isozymes.

Summary The contribution of different isozymes t o plasma alkaline phosphatase (AP) activity was investigated in a White PIymouth Rock strain. No significant difference in AP activity between FF and FS genotypes was observed in both young chick and laying hen. As previously reported in young chickens, a significant difference in AP activity between F and S types was observed in laying hens. Of the total variance of AP activity 53 %, 9 % and 5 7; were explained by isozyme type, family and sex, respectively. The higher activity of the F band was responsible for the higher activity of the F type in the young chicken, while the activity of the B band of either type did not contribute t o activity difference. The hypotheses are proposed so as to the activity difference.

A variant of the DR2 antigen, DR2 short (DR2S), was investigated in Japanese subjects by using a 9th International Histocompatibility Workshop reagent and locally-obtained alloantisera. A significant positive association between DR2S, Bw67 and TB23 was demonstrated. Haplotype analysis on four Japanese families showed that DR2S was segregated with the A11-Cw7-Bw67-DR2S-DQw1-TB23 haplotype.

Although the oxidative stress frequently occurs in patients with chronic hepatitis C, its role in future hepatocellular carcinoma (HCC) development is unknown. Hepatic 8-hydroxydeoxyguanosine (8-OHdG) was quantified using liver biopsy samples from 118 naïve patients who underwent liver biopsy from 1995 to 2001. The predictability of 8-OHdG for future HCC development and its relations to epidemiologic, biochemical and histological baseline characteristics were evaluated. During the follow-up period (mean was 6.7+/-3.3 years), HCC was identified in 36 patients (30.5%). Univariate analysis revealed that 16 variables, including 8-OHdG counts (65.2+/-20.2 vs 40.0+/-23.5 cells per 10(5) microm2, P0.0001), were significantly different between patients with and without HCC. Cox proportional hazard analysis showed that the hepatic 8-OHdG (P=0.0058) and fibrosis (P=0.0181) were independent predicting factors of HCC. Remarkably, 8-OHdG levels were positively correlated with body and hepatic iron storage markers (vs ferritin, P0.0001 vs hepatic iron score, P0.0001). This study showed that oxidative DNA damage is associated with increased risk for HCC and hepatic 8-OHdG levels are useful as markers to identify the extreme high-risk subgroup. The strong correlation between hepatic DNA damage and iron overload suggests that the iron content may be a strong mediator of oxidative stress and iron reduction may reduce HCC incidence in patients with chronic hepatitis C.

GB virus C (GBV-C) and hepatitis G virus (HGV) have been proposed as new viruses etiologically implicated in non-B, non-C hepatitis, but the morphology of these particular virus particles is still unknown, and most cases of non-A to E hepatitis do not relate to their infections. We tried to visualize virus-like particles (VLPs) in plasma samples from hepatitis B surface antigen- and antibody to hepatitis C virus (HCV)-negative blood donors with elevated alanine aminotransferase (ALT), and examined the association of the virus-like particles and the genomes of parenterally transmissible GBV-C/HGV. Twenty-three plasma samples, 13 with elevated ALT levels and 10 with normal ALT values, from blood donors without infections of hepatitis B virus (HBV) and HCV, were subjected to a 20%-60% sucrose density gradient centrifugation, and virus-like particles were observed by electron microscopy. GBV-C/HGV RNAs in the plasmas were tested. Virus-like particles were found in the fractions with densities of 1.15-1.16 g/ml from 12 of 13 (92.3%) plasmas with elevated ALT levels and 1 of 10 (10%) normal controls. The ultrastructural morphology of visualized VLPs was pleomorphic in size and appearance; the majority of the VLPs were 50- to 80-nm spherical particles with a 35- to 45-nm inner core and 9- to 12-nm-long surface spike-like projections. Rodlike VLPs 50-70 nm in diameter with a length of 110-160 nm were also observed in the same samples. The incidence of detection of the circulating VLPs was significantly (P < 0.001) related to elevated ALT levels, but GBV-C/HGV RNAs were detected in none of the plasmas containing the virus-like particles. Spherical VLPs are detected in HBV- and HCV-negative plasmas significantly correlated with the elevation of ALT, suggesting that they are implicated in non-B, non-C hepatitis.

Patients with chronic hepatitis C frequently have serum and hepatic iron overload, but the mechanism is unknown. Recently identified hepcidin, exclusively synthesized in the liver, is thought to be a key regulator for iron homeostasis and is induced by infection and inflammation. This study was conducted to determine the hepatic hepcidin expression levels in patients with various liver diseases. We investigated hepcidin mRNA levels of liver samples by real-time detection-polymerase chain reaction; 56 were hepatitis C virus (HCV) positive, 34 were hepatitis B virus (HBV) positive, and 42 were negative for HCV and HBV (3 cases of auto-immune hepatitis, 7 alcoholic liver disease, 13 primary biliary cirrhosis, 9 nonalcoholic fatty liver disease, and 10 normal liver). We analyzed the relation of hepcidin to clinical, hematological, histological, and etiological findings. Hepcidin expression levels were strongly correlated with serum ferritin (P < 0.0001) and the degree of iron deposit in liver tissues (P < 0.0001). Hepcidin was also correlated with hematological parameters (vs. hemoglobin, P = 0.0073; vs. serum iron, P = 0.0012; vs. transferrin saturation, P < 0.0001) and transaminase levels (P = 0.0013). The hepcidin-to-ferritin ratio was significantly lower in HCV(+) patients than in HBV(+) patients (P = 0.0129) or control subjects (P = 0.0080). In conclusion, hepcidin expression levels in chronic liver diseases were strongly correlated with either the serum ferritin concentration or degree of iron deposits in the liver. When adjusted by either serum ferritin values or hepatic iron scores, hepcidin indices were significantly lower in HCV(+) patients than in HBV(+) patients, suggesting that hepcidin may play a pivotal role in the pathogenesis of iron overload in patients with chronic hepatitis C.

Cell lines (2.2.15 cells) capable of supporting the replication of hepatitis B virus (HBV) DNA and intact viral particles have been established by HBV DNA transfection into HepG2 cells. The purpose of this study was to determine the ultrastructural morphology of native HBV particles without purification in the culture supernatants and in sera from patients. Electron microscopy (EM) and immunogold EM of the samples were carried out using polyclonal and monoclonal anti-hepatitis B surface antigen antibodies. HBV particles in the purified samples from the culture supernatants by density-gradient centrifugation were examined to compare the morphology with that of unpurified samples. EM and immunogold EM studies demonstrated the presence of Dane particles (41.8 nm in diameter), cobra-shaped (head diameter, 42.4 nm), and horn-shaped (head diameter, 43.5 nm) particles in the culture supernatants and in the sera from two patients. The tail of the cobra-like particles had a diameter of 21.0 nm and a length of 214 nm. The hornlike particles had a long branch 20.1 nm in diameter with a length of 189 nm, and a short branch 21.4 nm in diameter with a length of 112 nm. The ratio of Dane particles and cobra- and horn-shaped particles in the supernatants was 5 : 4 : 1. After ultracentrifugation, the cobra- and horn-shaped particles completely disappeared; there were only Dane particles together with spheres of 22 nm and filaments. In conclusion, this study showed for the first time that the native replicative form of HBV is cobra- and horn-shaped.

Lactoferrin is a milk protein with inhibitory effect on lipid peroxidation induced by oxidative stress. Oxidative stress plays an important role in the pathogenesis of chronic hepatitis C (CHC). The aim of this study was to evaluate the effect of bovine lactoferrin (bLF) monotherapy on lipid peroxidation, hepatic inflammation and iron metabolism in patients with CHC. Ninety Japanese patients with CHC were randomly assigned to two groups: bLF group (n=47) treated with bLF at a dose of 3.6g/day and a control group (n=43) that remained untreated. Plasma 8-isoprostane levels and clinical laboratory data including iron metabolism parameters were measured. Plasma 8-isoprostatne level was significantly decreased from 8.6+/-3.7 to 6.9+/-2.1pg/ml in the bLF group (P

We performed indirect immunogold electron microscopy (EM) for immunological identification and characterization of hepatitis C virus (HCV). To clarify the morphology of HCV, an indirect immunogold EM of two plasma samples from patients with high HCV RNA titers was carried out using antibodies specific for the putative HCV envelope protein (E) 1. Spherical virus particles 55-65 nm in diameter with delicate spike projections were detected in the 1.14-1.16 g/ml fractions after sucrose density gradient centrifugation. Polyclonal and monoclonal antibodies to the putative HCV E1 specifically recognized these particles. In addition, immunogold EM of the samples was also performed to uncover the morphology of HCV core particles. Spherical particles 33-40 nm in diameter (average, 37 nm) were detected in the 1.22- to 1.25-g/ml fractions by conventional EM after sucrose density gradient centrifugation. Immunogold EM using rabbit polyclonal antibody (RR8) specific for the putative HCV core protein and colloidal gold-labeled goat antirabbit IgG showed binding of the gold particles with RR8. Some of the HCV core particles showed icosahedric morphology. Optical rotation technique showed that the HCV core particles exhibit sixfold symmetry and that the length of the regular hexagon side is approximately 20 nm, suggesting that they have an icosahedric structure. Further, the detection limit of the indirect immunogold EM was evaluated in 11 plasma samples from chronic hepatitis B patients with different degrees of hepatitis B virus (HBV) DNA titers using antihepatitis B surface antigen antibody. The study showed that the detection limit of virus using this method is 10(7) virions/ml.

Background/Aims Nucleic acid damage by reactive nitrogen and oxygen species may contribute to inflammation-related carcinogenesis. To investigate the extent of nucleic acid damage in hepatitis C virus infection and its change after interferon treatment, we measured 8-nitroguanine and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in the liver of patients with chronic hepatitis C (CHC) before and after interferon therapy. Methods Hepatic accumulation of 8-nitroguanine and 8-OHdG was immunohistochemically evaluated in 20 CHC patients and 7 control patients with non-alcoholic fatty liver. Results Immunoreactivities of 8-nitroguanine and 8-OHdG were strongly detected in the liver from patients with CHC, but not in control livers. 8-Nitroguanine accumulation was found not only in infiltrating inflammatory cells, but also hepatocytes particularly in the periportal area. The accumulation of 8-nitroguanine and 8-OHdG increased with inflammatory grade (8-nitroguanine; P =0.0019, 8-OHdG; P =0.0009). In the sustained virological responder group after interferon therapy, 8-nitroguanine and 8-OHdG accumulation were markedly decreased in the liver (8-nitroguanine; P =0.018, 8-OHdG; P =0.018). Conclusions In this study, we demonstrated for the first time that 8-nitroguanine accumulated in the liver of patients with CHC. 8-Nitroguanine is a useful biomarker to evaluate the severity of HCV-induced chronic inflammation in relation to hepatocellular carcinoma.

Hepatitis C Virus (HCV) are 55-65 nm spherical particles, but the internal structure of the virion remains to be clarified. To clarify the morphology of HCV core particles, we performed an immune electron microscopy (IEM) using plasma samples from two blood donors with high HCV RNA titers and a detergent-treated anti-HCV core antibody-free plasma sample with high HCV RNA titer (1.5x10(8) copies/ml). Spherical particles, with 33-40 nm in diameter (an average diameter of 37 nm) were found in 1.22-1.25 g/ml fractions after sucrose density gradient centrifugation by conventional electron microscopy (EM). IEM using rabbit polyclonal antibody (RR8) specific to the putative HCV core protein and goat anti-rabbit IgG colloidal gold particles revealed that these spherical particles specifically reacted with RR8. This finding indicates that the spherical particles are naked HCV core particles. Some of the HCV core particles had an icosahedron-like structure. Optical rotation technique showed that the HCV core particle exhibits six-fold symmetry and that the length of regular hexagon side is approximately 20 nm. These findings showed that HCV core particles are spherical particles of 33-40 nm in diameter and that HCV core particles may possess an icosahedron-like structure and a buoyant density of 1.22-1.25 g/ml.

Background and aim Low-grade fasting hyperbilirubinemia is a common observation in healthy subjects (HS), whereas high-grade fasting hyperbilirubinemia is believed to be a characteristic finding of Gilbert's syndrome. This study was undertaken to assess the role of mutation in bilirubin UDP- glycosyltransferase gene (UGT1A1) on fasting hyperbilirubinemia. Methods Analysis of UGT1A1 and a caloric restriction test (400 kcal for 24 h) were performed in 56 healthy subjects (25 males, 31 females), and 28 patients with Gilbert's syndrome (18 males, 10 females). There were 29 healthy subjects with no mutation in UGT1A1, and 27 healthy subjects and 26 Gilbert's syndrome patients with mutations in the coding and/or promoter (TATA box) regions of UGT1A1. Results The mean increment of serum bilirubin (DeltaSB) was 7.6 micromol/L [corrected] (males) and 4.1 micromol/L (females) in subjects with no UGT1A1 mutation. Subjects with mutation in UGT1A1 showed higher levels of DeltaSB than individuals without mutation. Among healthy subjects, gender difference in DeltaSB values was observed only in individuals with the wild type of UGT1A1, but not in those with mutations in this gene. Conclusion The results of the present study suggest that UGT1A1 mutation has a role in the development of high-grade fasting hyperbilirubinemia after caloric restriction.

Divergent buoyant densities of hepatitis C virus (HCV) have been reported. If the destruction of HCV particles occurs during the ultracentrifugation process to separate fractions with different densities, an accurate evaluation of the HCV buoyant density is difficult. To examine this concern, changes were examined in HCV RNA titer of each density fraction after paraformaldehyde fixation of virus particles in the sera of 9 patients with chronic HCV infection. Serum was treated with 4% paraformaldehyde, and the density fractions were then separated by ultracentrifugation. The HCV RNA titer of each fraction was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay, and the results were compared with those obtained from the serum without paraformaldehyde fixation. After fixation, the HCV RNA titer was significantly increased in the 1.11-1.14 g/mL fraction (P=0.0018), and decreased in the 1.14-1.17 and 1.17-1.20 g/mL fractions (P=0.0457 and 0.0003, respectively). Using immunogold electron microscopy, it was found that morphologically destroyed HCV particles are present mainly in the 1.17 g/mL fraction of paraformaldehyde-untreated samples, whereas the intact HCV virion particles are present in the 1.12 and 1.14 g/mL fractions of the paraformaldehyde-treated samples. These results suggest that the destruction of HCV virions occurs during the ultracentrifugation process and that paraformaldehyde treatment protects from destruction. It was also considered that the accurate buoyant density of the HCV virion is 1.11-1.14 g/mL. This study describes a useful method for the purification of HCV virions, and provides new insights for elucidating the physicochemical properties of HCV particles.

The antitumor effects of a polyamine biosynthetic pathway inhibitor methylglyoxal bis(cyclopentylamidinohydrazone) (MGBCP) on the human hepatocellular carcinoma SK-HEP-1 cell line have been investigated. The growth of these cultured hepatocellular carcinoma cells was inhibited by MGBCP in a dose-dependent manner. Spermidine and spermine levels were dose-dependently depressed, and morphological changes due to programmed cell death (apoptosis) were observed in these MGBCP-treated hepatocellular carcinoma cells. These results suggest that in addition to reducing the growth rates, MGBCP can induce apoptotic cell death in this human hepatocellular carcinoma cell line.

In continuous billet casting, keeping the mold level steady is one of the most important technologies for maintaining steel quality. Using conventional methods, it is difficult to attain precise control of the mold level because of the nonlinear characteristics of the process. We have developed a control system using a neural network model to overcome this problem. In this paper, control problems of a continuous caster are introduced first. Next, the structure of the control system is proposed. In our proposed system, the neural network model recognizes the temporal patterns of inlet flow and controls the stopper stroke for a main control loop with a PI controller. The problems involved in construction of a valid neural network model that has good generalization and robust properties, are discussed from the viewpoint of optimizing the number of hidden layer units by the information criterion. Finally some results of its application are described.

The branched‐chain amino acid (BCAA)/tyrosine (Tyr) ratio (BTR) recently has been reported to be a good indicator of the severity of hepatic parenchymal injury in patients with chronic liver disease. In the present study, sequential changes of BTR after BCAA administration were determined in patients with chronic liver disease to evaluate the value of BTR as a marker of the clinical response to nutritional therapy in these patients. This study comprised 75 patients with chronic hepatitis and 96 with liver cirrhosis. BTR was significantly decreased in patients with cirrhosis and hepatitis compared with healthy subjects. BTR was significantly correlated with the Child‐Pugh score and with other liver function tests. BCAA increased significantly 2 hr after BCAA administration and decreased gradually thereafter. Tyr significantly decreased 4 hr after BCAA administration. BTR significantly increased 2 and 4 hr after BCAA therapy. The increase in BTR 3 hr after BCAA administration was low in patients with decreased basal BTR. The results of this study showed that BTR is a good index of the hepatic parenchymal damage and that it may be a useful marker for monitoring response to nutritional therapy in patients with chronic liver disease. J. Clin. Lab. Anal. 13:31–34, 1999. © 1999 Wiley‐Liss, Inc.

The magnetisation transfer ratio (MTR), a quantitative tissue characteristic, and the pallidal index were measured in the globus pallidus of 37 patients with hepatic cirrhosis and 37 control subjects. The MTR decreased with severity of the disease, as indicated by the serum total bililubin, indocyanine green 15-min retention rate, and plasma ammonia. The pallidal index correlated significantly with haematological parameters, although the correlation coefficients tended to be lower than those between MTR and haematological parameters. This change in MTR corresponded to the results of the manganese chloride experiments. T1 shortening in the pallidum of patients with cirrhosis is presumably caused by paramagnetic effects, and possibly attributed to manganese deposition.

Interleukin‐6 receptors (IL‐6R) and interleukin‐1 receptors (IL‐1R) on lymphocyte surfaces were analyzed, using flow cytometry and dye‐labeled IL‐6 and IL‐1β, to examine the clinical and immunological significance of these receptors. Incubation of peripheral blood mononuclear cells in the presence of mitogen resulted in a remarkable increase of lymphocytes expressing the IL‐6 and IL‐1β receptors on the cell surface. The increase in lymphocytes bearing these cytokine receptors may reflect an increase in stimulated lymphocytes. When peripheral blood from patients with primary biliary cirrhosis (PBC) was examined for these receptors, the percentage of IL‐6R positive cells was significantly higher in the patients than in healthy controls (P

The effect of 6 h transportation on plasma levels of catecholamines, cortisol, glucose and the number of eosinophils before, during and after the transportation was studied in adult male Japanese native Tokara goats. Blood was obtained via indwelling catheters in the jugular and femoral veins. During transportation, various behaviors apparently indicated nervousness or excitement. Plasma epinephrine levels increased immediately after the start, from a base value of 7.5 pmol ml −1 to 25.9 pmol ml −1 ( P −1 before transport and rose immediately after the start to a peak value of 166 ng ml −1 ( P −1 before the start to 165 mg dl −1 ( P −1 ( P

合鴨農法における家鴨類の適性品種を選定するため,マガモ,チェリバレーおよびF1(マガモ×チェリバレー)の3品種の行動および除草•防虫能力について比較検討した。試験田を3つの放飼区(マガモ,チェリバレーおよびF1)に分け,田植え1週間後にそれぞれ3品種の0日齢の雛を放飼し,放飼期間は稲の出穂期までの11週齢とした。結果を要約すると以下のとおりである。1. 水田内における労働時間(摂食行動と移動時間の合計)は,マガモが最も長く,次いでチェリバレー,F1の順であった。2. 水田内における休息時間は,F1,チェリバレーおよびマガモの順に長い傾向を示した。3. 1時間当たりの摂食行動回数は,マガモがF1およびチェリバレーに比較して多い傾向を示した。4. 水田内の発生状況調査から見た家鴨類の除草および防虫能力には,3品種間に有意な差は認められなかった。

合鴨水稲作における家鴨類の適性品種を選定するため,マガモおよびチェリバレーの2品種とそれらの交雑種(F1)の繁殖能力について検討した。試験区を3つの区(マガモ,チェリバレーおよびF1)に分け,マガモ16羽(_??_13:_??_3),チェリバレー15羽(_??_12:_??_3)およびF120羽(_??_16:_??_4)を供試して,1区当たり約30m2の運動場に放し飼いした。試験期間は1994年6月16日(0日齢)から1995年8月1日(411日齢)までの約14ヵ月であった。試験開始時から終了時まで2品種および交雑種とも同様に,成鶏用配合飼料を不断給与し,産卵開始時から試験終了時まで週1回飼料摂食量を計った。結果を要約すると以下のとおりである。1. 171～410日齢までの平均産卵率については,チェリバレーとF1に比べてマガモの産卵率が明らかに低いことが認あられ,卵重はチェリバレー,F1,マガモの順に大きいことが認められた。2. 171～410日齢までの1日1羽当たり飼料消費量は,チェリバレー,F1,マガモの順に多いことが認められた。飼料要求率については,チェリバレーとマガモおよびF1とマガモの間に有意差はなかったが,チェリバレーとF1の間に有意差が認められた。3. ハウユニットについては,2品種および交雑種の間には有意差が認められなかった。卵形係数卵黄係数および卵殻の厚さでは,チェリバレーはF1とマガモに比べて優れており,卵白pHについては,チェリバレーとF1は同程度であったが,マガモは有意に高い値を示した。4. チェリバレーとF1に比べてマガモの受精率はかなり低いことが認められた。孵化率については,チェリバレーとF1との間に有意差が認あられなかったが,これらに比べてマガモがかなり低いことが認められた。5. 以上の結果から,2品種間および交雑種ではチェリバレーが家鴨類の繁殖能力から見た適性品種であることが示唆された。

合鴨農法における家鴨類の適性品種を選定するため,マガモ,チェリバレーおよびF1(マガモ×チェリバレー)の3品種の成長および産肉能力について比較検討した。試験田を3つの放飼区(マガモ,チェリバレーおよびF1)に分け,田植え1週間後にそれぞれ3品種の0週齢の雛を放飼した。放飼期間は稲の出穂期までの11週齢とし,その後20週齢まで舎飼肥育して屠殺解体し,産肉諸形質について検討した。なお,水田放飼区の対照区として,それぞれの3品種につき配合飼料単一給与の舎飼区を設けた。1. 20週齢終了時における3品種の家鴨類の体重は,水田放飼区,舎飼区ともにチェリバレー,F1およびマガモの順に大きい傾向を示した。2.0～20週齢の水田放飼区における家鴨類の成長は,舎飼区に比較してチェリバレーが低い傾向を示したが,F1とマガモは舎飼区とほぼ同様の傾向を示した。3. 枝肉歩留りは,3品種間ならびに水田放飼区と舎飼区の間に有意な差は認められなかった。腹腔内脂肪はチェリバレーが最も多く,次いでF1,マガモの順に多い傾向を示した。4. 3品種の家鴨類の食味テストの結果,F1,マガモおよびチェリバレーの順に嗜好性が高い傾向を示した。5. 以上の結果から,3品種のうちではF1が水田放飼に最も適した品種であることが示唆された。

The noninvasive determination of effective hepatic blood flow, intrahepatic shunted blood flow, intrahepatic shunt index, and total hepatic blood flow was investigated by using the sequential single photon emission computed tomography. This method was performed for a period of 10 minutes following an intravenous injection of 99mTc-(Sn)-N-pyridoxyl-5-methyltryptophan and a venous blood sampling. This study comprised 8 healthy volunteers, 16 patients with chronic hepatitis, and 33 patients with liver cirrhosis. The intrahepatic shunt index measured with this method coincided with the intrahepatic shunt index determined by catheterization, indicating the high reliability of this procedure. The effective hepatic blood flow in patients with liver cirrhosis was significantly lower than that in the healthy controls and the chronic hepatitis group. The intrahepatic shunted blood flow was significantly higher in patients with liver cirrhosis compared with the flow in healthy controls. The intrahepatic shunt index was also significantly higher in patients with liver cirrhosis compared with the index of healthy controls and those with chronic hepatitis. No substantial differences were noted in the total hepatic blood flow among the three groups. The effective hepatic blood flow, the intrahepatic shunted blood flow, and the intrahepatic shunt index, correlated with the serum albumin concentration, the serum cholinesterase level, and the plasma indocyanine green attenuation rate. From these results, it was concluded that the present procedure constitutes a reliable and effective method for the noninvasive determination of hepatic blood flows. Consequently, it will be of high clinical value for assessing the functional and the pathological alterations of the liver.

To clarify the morphology of hepatitis C virus (HCV), an indirect immunogold electron microscopic study was carried out on two plasma samples with high HCV RNA titres using polyclonal and monoclonal antibodies specific to the putative HCV envelope protein. Spherical virus-like particles, 55 to 65 nm in diameter with spike-like projections, were found in 1.14 to 1.16 g/ml fractions after sucrose density gradient centrifugation. These particles were found only in HCV-infected blood donors and had morphological features similar to those of flaviviruses. Moreover, these particles specifically reacted with the polyclonal and monoclonal antibodies to the putative HCV envelope protein. This is the first known report in which the morphology of the HCV particle is clearly shown.

輸送開始時のストレッサーに対する反応に知覚-心理的経路が関与する可能性を明らかにするため,頸静脈と股静脈に残置カテーテルを装着した雄トカラヤギ3頭を用いて,ブランコ実験と実際の路上輸送試験を行なった.ブランコ実験ではケージにいれたままの自然の振動を組み合せた1セット15分間の振動を6回繰り返した.実際の輸送試験ではケージに入れたヤギをトラックに乗せて公道を3時間走った.ブランコ振動の間ヤギはあわててくずれた姿勢を直そうとし,また落ち着きのない行動を示した.ブランコ振動が始まるとすぐに,不規則で激しい振動のときヤギは急性で一過性の心拍数増加を示した.これと対照的に呼吸数はブランコ振動と関係がみられず,実際の経過時間と関係していた.ブランコ実験開始後30分には,血漿中のコルチゾールとグルコースがそれぞれ基礎値の4倍と3倍に上がっていた.プランコ運動終了時にはコルチゾールは同じく4倍に上昇した.そして,これらはプランコ終了後3時間で基礎値にもどった.ブランコによって好酸球は明らかに減少し開始後4.5時間で最小値を示した.白血球数は振動開始後漸増し,振動終了後3時間と12時間にピークを示した.トラック輸送では反応の大きさと持続時間がより明らかであったが,コルチゾールとグルコースは本質的にブランコ実験と同じ反応を示した.好酸球の減少は2頭にみられた.白血球数は輸送開始時から増加し,2時間後にピークを示した.コルチゾール,グルコースおよび白血球数の増加と好酸球数の減少のタイミングは,緊密に同期して実験開始直後から起っており,また,これらの反応はプランコ実験と実際の輸送試験の両方に認められた.今回の試験開始時における,実験動物の行動,コルチゾール,血液学的反応は,知覚-心理的経路の関与によって,動物が未知の処置と環境に対する恐怖と不安のために,驚愕反応を起しているにとを示唆している.

塩味(塩化ナトリウム),酸味(酢酸),甘味(ショ糖),苦味(カフェイン),旨味(グルタミン酸ナトリウム)およびアルコール(エタノール)に対する牛の味覚反応を2瓶選択法で調べた.これらの味物質を水道水に溶かしてそれぞれ7段階の濃度を設定し,低濃度から高濃度へ向けて実験を行なった.各濃度における総飲水量に対する試験液の摂取割合を選好指数とし,濃度上昇に伴う選好指数の推移を味覚反応曲線として表し,味覚反応を推察した.χ2-検定により,選好指数39.7%以上から60.3%未満は不弁別範囲,60.3%以上は選好範囲,39.7%未満は拒絶範囲と定義した.塩味物質は濃度1.25%で弱い拒絶反応を示した以外はほとんど水と弁別されなかった.牛は酸味物質に対して全体的に拒絶反応を示し,特に濃度0.08%以上では強い拒絶反応を示した.甘味物質は濃度0.16%から1.25%まではほとんど水と弁別されなかったが,2.5%以上では嗜好を示し,5.0%で最も強い嗜好を示した.苦味物質は濃度0.01%で弱い嗜好を示した以外はほとんど水と弁別されなかった.旨味物質は濃度0.08%以上では嗜好を示し,濃度0.63%で最も強い嗜好を示した.牛はアルコール物質に対して濃度1.6%で弱い嗜好を示した以外はほとんど水と弁別せず,濃度の上昇ににつれて強い拒絶反応を示した.

We quantified serum hepatitis C virus RNA titers and determined hepatitis C virus subtypes in chronic hepatitis C patients treated with interferon-β to investigate relationships among serum ALT response, serum hepatitis C virus titer and hepatitis C virus subtype. Of 146 chronic hepatitis C patients who received interferon-β therapy, 24 patients with sustained serum ALT normalization (complete responders) and 26 patients without serum ALT normalization (nonresponders) were randomly selected. Detection, typing and quantitation of hepatitis C virus were performed by means of the “single-tube” polymerase chain reaction method. Of the 24 complete responders, 21 (87.5%) became negative for hepatitis C virus RNA, whereas 21 (80.8%) of the 26 nonresponders remained positive. Hepatitis C virus infections with types I, II, III, IV, II + III and III + IV occurred in 0 (0%), 22 (51.2%), 10 (23.3%), 1 (2.3%), 7 (16.5%) and 3 (7.9%) patients, respectively. The mean pretreatment hepatitis C virus RNA titer of complete responders (0.4 ± 2.0 × 104 CID50/ml) was significantly lower than that of nonresponders (3.8 ± 4.5 × 104 CID50/ml) (p < 0.01). Regardless of HCV subtype, patients with more than 104 CID50/ml of HCV did not show serum ALT normalization, whereas complete serum ALT response was seen in most cases with less than 102 CID50/ml HCV. These results show that mixed infections with different hepatitis C virus subtypes appear to be more common than previously reported and that the pretreatment serum level of hepatitis C virus RNA is a more important predictor of outcome of interferon therapy than is virus genotype. (HEPATOLOGY 1993;18:1319–1325.)

We have developed a new technique to quantify hepatitis C virus (HCV)-RNA using polymerase chain reaction (PCR) and dot blot hybridization. PCR was performed on 10-fold dilutions of an HCV-cDNA fragment for various numbers of cycles and on various serum volumes for 30 cycles. The resulting PCR products were spotted onto nitrocellulose filters and hybridized with a 32 P-labelled internal probe. The intensity of the hybridization signal was proportional to the quantity of template cDNA and increased in proportion to the number of PCR cycles. Clear signals were detected after 30 cycles of PCR using an original serum volume of greater than 250 pl. Thus, this new technique allows the quantitation of HCV-RNA, and as it is simple and cost-effective, it promises to have many clinical applications.

The antiviral effect of natural interferon (IFN)-α on chronic hepatitis C virus (HCV) infection was estimated by determining quantitative changes in serum HCV-RNA compared with the serum alanine aminotransferase (sALT) improvement; the relationships of responses to IFN according to the dose and period of IFN therapy were defined to determine an appropriate IFN therapy protocol. Twenty-two patients with chronic hepatitis C were given natural IFN-α and in 16 (72.7%) patients the viraemia was suppressed during therapy. Five (27.7%) of them sustained the disappearance of HCV-RNA for more than 6 months after therapy accompanied with a prolonged sALT improve-ment. Pre-treatment viraemia levels in 5 complete responders with "complete suppression" of viraemia were significantly lower than in 11 patients with a transient loss or a decline of HCV-RNA. Afavorable antiviral response was closely associated with a high total dose of IFN-α and a long duration of IFN therapy.(Internal Medicine 32: 523-529, 1993)

合鴨の水田放飼が稲の生育および収量に及ぼす影響について検討した。試験田を無農薬•合鴨無放飼の対照区5a,農薬散布区(以下農薬区に略)15aおよび合鴨放飼区(以下合鴨区に略)10aの3区に分け,田植え2週間後に4週齢の合鴨の雛35羽を放飼した(最終的には17羽に調整)。放飼期間は稲の出穂期までの約2ヵ月間とした。1.稲の草丈は農薬区で前半期に高い傾向を示したが,後半期では3区間に差は認められなかった。2.稲の茎数は多い順に合鴨区,対照区,農薬区であった。3.稲の葉面積は農薬区が最も大きく,次いで合鴨区,対照区の順となった。また稲の高度別の相対照度は,草丈60～70cmでは3区間に差異は認められなかったが,草丈50cm以下では農薬区で低い傾向を示した。4.合鴨区の水田内の欠株数は446株で欠株率は4.5%を示した。5.1株穂数,1株登熟粒数および1株収量は,合鴨区,対照区,農薬区の順に多い傾向を示した。しかしながら10a当たり収量では,合鴨区と農薬区では有意な差は認あられなかった。

水田内に放飼した合鴨の成長と行動について検討した。試験田10aを用い,田植え2週間後に4週齢の合鴨30羽を放飼した。一方,22羽をブロイラー用後期配合飼料で舎飼し対照区とした。水田放飼区と舎飼区の合鴨の体重,脛長,嘴長および体長を毎週1回測定した。また,水田放飼区の合鴨群から5羽を任意に抽出し,行動型を24時間にわたって調査した。終了時の体重は水田放飼区976g,舎飼区970gを示し,1日当たり増体量はそれぞれ8.3gおよび8.2gとなり,両区間に有意な差は認められなかった。また,終了時の体長,脛長および嘴長等のいずれの部位も両区間で有意差は認められなかった。24時間の採食行動は14時～16時,9時～11時および2時～7時の3つのピークが見られたが,中でも2時から7時までの採食行動が最も大きなピークを示した。休息行動は8時～10時と17時～2時の2つのピークが見られたが,中でも17時から2時までの休息行動が最も大きなピークを示した。以上のことから,合鴨の一日の行動パターンは野鴨と同様に夜行性であることが明らかとなった。

塩味(塩化ナトリウム),酸味(酢酸),甘味(ブドウ糖),苦味(塩酸キニーネ),旨味(グルタミン酸ナトリウム)に対する鶏•鶉の味覚反応を2瓶選択法で調べた.これらの味物質を水道水に溶かしてそれぞれ11段階の濃度を設定し,低濃度から高濃度へ向けて実験を行なった.各濃度における総飲水量に対する試験液の摂取割合を選好指数とし,濃度上昇に伴う選好指数の推移を味覚反応曲線として表し,味覚反応を推察した.X2-検定により,選好指数39.7%から60.3%は不弁別範囲,60.3%以上は嗜好範囲,39.7%以下は拒絶範囲と定義した.塩味の濃度は鶏が0.64%,鶉が1.28%まで弁別せず,鶏が1.28%,鶉が2.5%以上で拒絶反応を示した.酸味の濃度は鶏が0.04%,鶉が0.16%まで弁別せず,鶏が0.08%,鶉が0.32%以上で拒絶反応を示した.甘味の濃度は鶏が20%まで弁別せず,鶉は1.28%と2.5%で弱い嗜好を示した.苦味の濃度は鶏が0.02%まで弁別せず0.04%以上で拒絶反応を示し,鶉は0.32%まで弁別しなかった.旨味の濃度は鶏が0.64%,鶉が5%まで弁別せず,鶏が1.28%以上,鶉が10%で拒絶反応を示した,以上の結果より,鶏と鶉は特に強い嗜好を示す味物質はないことと,鶏は鶉より味覚が敏感であることが明らかとなった.

特発性急性妊娠脂肪肝(Idiopathic acute fatty liver of pregnancy, IAFLP)の1救命例を経験し,その経時的な肝容積の変化と肝組織所見とを比較検討したので報告する.症例は36歳3回経妊3回経産婦.妊娠38週より黄疸,意識障害が出現.男児を分娩後,当科に緊急入院となった.CTスキャンによる肝容積は第2病日に約703cm3と減少を認めたが,肝機能の改善とともに第8病日には約1,390cm3と急速に正常化した.第12病日の肝組織所見では,肝細胞の壊死・脱落は軽微であり,肝容積の変化はおもに,個々の肝細胞の萎縮から腫大への変化に起因するものと考えられた.IAFLPの経過予後の判定に肝CTスキャンによる経時的な肝容積の測定は有用であった.

Many patients with porphyria cutanea tarda (PCT) have been reported to be hepatitis C virus (HCV) carriers, suggesting that HCV infection plays a role in the pathogenesis of this type of porphyria. In this study, we report a patient with chronic hepatitis C-associated PCT. Therapy with interferon (IFN) transiently decreased HCV RNA levels, but levels of urinary porphyrins and serum transaminases and ferritin remained unchanged. Serum ferritin and urinary porphyrin levels improved after phlebotomy, but this therapy was not effective in improving serum transaminase levels. Although a physiopathological association between HCV infection and PCT has been suggested previously, IFN was not effective in this patient. The transient decrease in HCV RNA levels was a factor independent of porphyrin metabolism.

Several lines of evidence have suggested that oxidative stress plays an important role for the pathogenesis of nonalcoholic steatohepatitis (NASH). Therefore, by using immunohistochemical staining of liver biopsy samples, we measured hepatic 7,8-dihydro-8-oxo-2′ deoxyguanosine (8-oxodG), a DNA base-modified product generated by hydroxyl radicals, of 38 NASH patients and compared with 24 simple steatosis and 10 healthy subjects. Relation of hepatic 8-oxodG with clinical, biochemical, and histologic variables and changes after iron reduction therapy (phlebotomy plus iron–restricted diet) were also examined. Hepatic 8-oxodG levels were significantly higher in NASH compared with simple steatosis (17.5 versus 2.0 8-oxodG–positive cells/105 μm2; P < 0.0001). 8-oxodG was significantly related to iron overload condition, glucose-insulin metabolic abnormality, and severities of hepatic steatosis in NASH patients. Logistic regression analysis also showed that hepatic iron deposit and insulin resistance were independent variables associated with elevated hepatic 8-oxodG. After the iron reduction therapy, hepatic 8-oxodG levels were significantly decreased (from 20.7 to 13.8 positive cells/105 μm2; P < 0.01) with concomitant reductions of serum transaminase levels in NASH patients. In conclusion, iron overload may play an important role in the pathogenesis of NASH by generating oxidative DNA damage and iron reduction therapy may reduce hepatocellular carcinoma incidence in patients with NASH. (Cancer Epidemiol Biomarkers Prev 2009;18(2):424–32)