Kiyowo Kosaka

Lectin-Reactive Profiles of Alpha-Fetoprotein Characterizing Hepatocellular Carcinoma and Related Conditions

Serum alpha-fetoprotein from 146 patients with hepatocellular carcinoma, other malignancies, and benign liver diseases, was fractionated by lectin-affinity electrophoresis coupled with our sensitive detection method of antibody-affinity blotting. Compared with chronic hepatitis and liver cirrhosis, hepatocellular carcinoma was characterized by the increase in proportions of lentil lectin A-reactive alpha-fetoprotein-L3 and erythroagglutinating phytohemagglutinin-reactive alpha-fetoprotein-P4; the yolk sac tumor was characterized by the increase of concanavalin A-nonreactive alpha-fetoprotein-C1, lentil lectin-A-weakly reactive alpha-fetoprotein-L2, erythroagglutinating phytohemagglutinin-strongly reactive alpha-fetoprotein-P5, and Allomyrina dichotoma lectin-nonreactive, slow-migrating alpha-fetoprotein-Als; and gastrointestinal tumors were characterized by alpha-fetoprotein-C1, alpha-fetoprotein-L2, alpha-fetoprotein-L3, alpha-fetoprotein-P5 and Allomyrina dichotoma-nonreactive alpha-fetoprotein-A1. By combined evaluation of alpha-fetoprotein-L3 and alpha-fetoprotein-P4, hepatocellular carcinoma was discriminated from chronic hepatitis and liver cirrhosis with a sensitivity of 97% at a specificity of 99.7%. Because the alpha-fetoprotein level of the studied cases ranged from 60-1,500,000 ng/mL (60-1,500,000 micrograms/L), mostly greater than 200 ng/mL (200 micrograms/L), additional patients with lower levels of alpha-fetoprotein [16-177 ng/mL (16-177 micrograms/L) for 16 cases of hepatocellular carcinoma with liver cirrhosis and 28-185 ng/mL (28-185 micrograms/L) for 17 cases of liver cirrhosis alone] were analyzed for alpha-fetoprotein-L3 and alpha-fetoprotein-P4. The resulting sensitivity for combined evaluation was still as high as 88% at the same high specificity of 99.7%, indicating that the simultaneous analysis of alpha-fetoprotein-L3 and alpha-fetoprotein-P4 is effective in monitoring the evolution of hepatocellular carcinoma in cirrhotic patients.

MICROHETEROGENEITY OF RAT α-FETOPROTEIN*

A purified and homogeneous preparation of rat AFP, as judged by both electrophoresis on Cellogel and immunoelectrophoresis, was separated into two components, AFPa and AFPb, by disc electrophoresis on 7% polyacrylamide gel. These two components had a definite difference in electrostatic net charge and gave only a single band on SDS-electrophoresis. Immunological reactivity or electrophoretic separation or mobility of the two components could be altered by treatment with either sulfhydryl inhibitors or reducing agents but not by treatment with protein denaturants. Electrophoresis of neuraminidase-treated AFP on 5% polyacrylamide gel yielded clearly separable, slower moving four to six and finally two components depending on the time of incubation with neuraminidase. The time-dependent conversion of faster into slower migrating components of both AFPa and AFPb upon neuraminidase treatment was confirmed by reelectrophoresis of separated and similarly treated AFPa and AFPb. Two bands of sialized or desialized AFP were also observed on isoelectric focusing. Both AFPa and AFPb treated with and without neuraminidase gave single fused precipitin lines against the antiserum in Ouchterlony double-diffusion analysis. On the basis of the changes in electrophoretic mobilities of the intermediates following neuraminidase treatment, AFPa and AFPb were estimated to have at least 2.5 and 4.5 molecules of sialic acid per molecule, respectively.

DIFFERENT MECHANISMS OF INCREASED α‐FETOPROTEIN PRODUCTION IN RATS FOLLOWING CCl4 INTOXICATION AND PARTIAL HEPATECTOMY

The temporary increase in serum a-fetoprotein ( AFP) concentration during the course of hepatitis and cirrhosis of the liver in man has been suggested to be caused by liver regeneration. This suggestion is based on a similar kinetic alteration of AFP level observed in mice and rats with regenerating liver.*v2 In these experimental animals, regenerating livers have been produced by either carbon tetrachloride (CCI,) intoxication or partial hepatectomy. This complicates the interpretation of the experimental results, because CCI, poisoning causes not only regeneration but also degeneration and necrosis of hepatic cells. Our studies of key carbohydrate-metabolizing enzymes in rat livers after CCI, injury or partial hepatectomy have revealed that the extent of enzyme deviation in the injured liver is much greater than that in the liver after partial hepatectomy and that the marked enzyme alteration in the injured liver is caused by hepatic cell injury per se and is not the result of liver regeneration.3 With this view in mind, we undertook the present study to make a direct comparison of serum AFP levels following CCI, intoxication and partial hepatectomy of rats. The effects of mitomycin C and 8-azaguanine on the AFP level after these treatments were also studied to correlate the AFP production with the syntheses of DNA and RNA.

Differential responses of glucose 6-phosphate dehydrogenase and lipogenesis to fat-free diets

Abstract Fasting and refeeding rats on a glucose-casein diet increased both the activity of glucose 6-phosphate dehydrogenase and the conversion of glucose into fatty acids in liver, whereas refeeding on glucose alone caused a comparable rise in the lipogenic rate without altering the dehydrogenase activity, indicating that the induced levels of the dehydrogenase are not necessarily required for the increased generation of NADPH under the adaptive hyperlipogenesis. This was partially proved by the finding that the oxidation of carbon 1 of glucose was preferentially increased over carbon 6 in the liver of glucose-refed rats. The activities of malic enzyme, citrate cleavage enzyme and glucokinase were elevated substantially by refeeding glucose with or without added protein. These results were interpreted as indicating that the response of enzymes catalyzing the carbon chain-supplying reactions in fatty acid synthesis from glucose is of a primary importance for the adaptive hyperlipogenesis and that the induction of glucose 6-phosphate dehydrogenase by the glucose-casein diet is rather representing a metabolic consequence secondary to the enhanced fatty acid synthesis.

Pancreatic-type hyperamylasemia in end-stage renal disease.

Pancreatic-type isoamylase (P-type) and salivary-type isoamylase (S-type) activities were determined by the wheat protein inhibitor method in 29 patients with end-stage renal disease and in 38 healthy volunteers. Serum levels of total amylase (322±43 units/liter) and P-type (212±39 units/liter) in ESRD were significantly higher than those of controls (total: 142±7 units/liter,P<0.01; P-type: 52±4 units/liter,P<0.01). There was no significant difference between S-type activities in ESRD (110±16 units/liter) and in controls (90±6 units/liter). The ratios of amylase clearance to creatinine clearance (Cam/Ccr) and S-type clearance to creatinine clearance (Cs-amy/Ccr) rose significantly in ESRD(Cam/Ccr: 5.7±0.6%; Cs-amy/Ccr: 4.3±0.55%) compared to controls (Cam/Ccr: 3.2±0.24%,P<0.01; Cs-amy/Ccr: 2.1±0.17%,P<0.01). The ratio of P-type clearance to creatinine clearance (Cp-amy/Ccr) revealed no significant difference between ESRD (5.5±0.54%) and controls (5.6±0.42%). The renal excretion of P-type appeared to be more impaired than that of S-type in ESRD.

Intranuclear Virus-Like Particles in Hepatocytes of an Asymptomatic Hepatitis B Antigen Carrier with Dane Particles in the Serum

An asymptomatic hepatitis B antigen carrier with Dane particles in the serum and intranuclear virus-like particles in hepatocytes is reported.

Prevention of Choline-Deficiency Fatty Liver of Rats with Xylitol

Fatty liver and cirrhosis produced in rats maintained on a diet low in protein and choline can be modified by the amount and kind of fat included in the diet [1–4]. Recent advances in clinical nutrition, notably carbohydrate [5, 6], prompted us to investigate the effect of changing the carbohydrate in the diet from sucrose to others. In the present communication, it will be reported that the production of fatty liver is modified by the substitution of sucrose for xylitol. In addition a certain unexpected effect of xylitol that might be related to the action of choline will also be described.

Parotid swelling. Does it affect serum isoamylase level

Serum levels of total amylase and P-type, and S-type isoamylase were measured in 57 patients with chronic parotid swelling (41 men and 16 women; mean age 60.4) and the same number of age- and sex-matched control individuals without parotid swelling (41 men and 16 women; mean age 60.3). None of the three enzyme measurements differed significantly between patients with parotid swelling and control subjects. It is concluded that chronic enlargement of parotid glands does not affect serum isoamylase level.

Two cases of secondary hyperparathyroidism that showed marked enlargement of jaw bones.

長期血液透析療法中に顎骨の膨隆により, 顔貌の高度な変化をきたした2症例を経験したので報告する. 症例1は透析歴10年の48歳男性, 透析7年目頃より踵部に始まる骨痛が出現, その後, 手指先端の腫大, 円背, 胸郭の狭小化とともに顎骨の膨隆が現われ, 以後, 増大し, 顔貌の高度な変化をきたした. 上顎骨生検は線維性骨炎の像を呈し, AI-P 84.0 KAU, C-PTH 76.8ng/ml以上と高値で, 画像上も副甲状腺腫大を認めた. 2°HPTと診断し, 副甲状腺摘出術を施行した結果, 顎骨膨隆は軽度改善傾向がみられている. 症例2は透析歴8年の31歳女性, 透析5年目頃より, 足底部に始まる骨痛が出現, 以後, 症例1と同様の症状を伴って顎骨の膨隆が現われ, 顔貌の変化をきたした. AI-P 80.3 KAU, C-PTH 76.8ng/ml以上と高値を示し, 画像上も副甲状腺腫大を認めた. 両例ともにCa, Pのコントロールが非常に悪く, 本症発症との関連が示唆された.

HCV infections in patients on hemodialysis. Mainly on the infection routes by retrospective study.

透析施設の今日的課題であるC型肝炎の実態とその感染経路について検討した. Ortho社製ELISAキットを用いてC型肝炎ウイルス (HCV) 抗体を測定した結果, 血液透析患者102名中24名 (23.5%) がHCV抗体陽性であった. 凍結保存血清を用いた検索にて, 陽性者24名中11名は当院透析開始 (以後, 開始) 時にはすでに陽性で, 13名は開始後陽性化していたことが判明した. 開始後陽性化群13名と持続陰性群77名との比較では透析期間は陽性化群が有意に長かったが, 輸血歴および輸血量に関しては両者に有意差はなかった. 開始時陽性者は陰性者に比して, 開始前輸血歴を有する者が有意に多かったが, 開始後陽性化者と持続陰性者との間には輸血歴に関して有意差はなかった. 開始後陽性化した13名中3名には関連輸血歴はなく, 開始後輸血歴のない全患者15名の20%にあたり, わが国の供血者の陽性率より高率であった. 以上, 透析患者はHCV感染のhigh risk groupである. その感染経路は輸血によるものが多かったものの, 開始後に輸血以外の感染経路も存在することが示唆された. 従って, 透析施設内の感染予防対策の確立が必要であると考えられた.