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The characteristics of two monoclonal antibodies to poly(ADP-Rib) and the various structures of poly(ADP-Rib) recognized by these monoclonal antibodies have been examined. One antibody, IgG monoclonal antibody 10H, reacted with most parts of poly (ADP-Rib) molecules, and its binding was only slightly inhibited by Ado(P)-Rib-P, a monomer unit of the polymer. On hydrolysis of poly(ADP-Rib) protected by the antibody hydrolytic products, such as Ado(P)-Rib-P and Ado(P)-Rib(P)-Rib-P, were almost the same as those from control poly(ADP-Rib). The other antibody, IgM monoclonal antibody 16B, recognized only some parts of poly(ADP-Rib) molecules of larger size, and its binding was not inhibited by Ado(P)-Rib-P. Branched portions were considerably concentrated in parts protected by this antibody. These two monoclonal antibodies are suggested to recognize different structures of poly(ADP-Rib): possibly IgG antibody 10H recognizes linear portions whereas IgM antibody 16B recognizes branched portions of the polymer. These monoclonal antibodies should be useful in studies on the structures, including unknown structures and the locations of these structures in poly(ADP-Rib).

Phosphatidylcholine (PC), especially dilinoleoyl-PC, has been reported to be effective in preventing hepatic fibrosis in chronically alcohol-fed baboons. Continuous hepatic inflammation predisposes the structure of the liver to fibrosis. Since n-3 polyunsaturated fatty acids (PUFA) have been shown to exhibit an anti-inflammatory effect, we tested the hypothesis that n-3 PUFA PC as a dietary supplement has a beneficial effect on chronic liver disease susceptible to fibrosis. Salmon roe phospholipids, 90% of which are PC, were extracted and encapsulated. Almost a third of the PC fatty acids were docosahexaenoic acid (22:6 n3) and 10% were eicosapentanoic acid (20:5 n3). About 1600 mg/day of the phospholipids was administered for six months to six chronic liver disease patients, four with hepatitis B infection (three with cirrhosis, one with chronic hepatitis), one with hepatitis C virus cirrhosis and one with alcoholic cirrhosis. There was no change in the results of blood chemistry studies related to liver function, except in globulin, which decreased from 3.80 g/dl to 3.67 g/dl (p < 0.05). Among the lipid parameters, HDL-cholesterol, apolipoprotein A-I and apolipoprotein E increased significantly. Although this was a small trial, n-3 PUFA PC may be beneficial in the treatment of chronic liver diseases.

症例は58歳,男性.咽頭炎発症後,左肩痛が出現し,全身倦怠感のため入院となった.左鎖骨周囲にガス壊疽を来し, CT検査で骨髄炎,肝膿瘍,敗血症性肺塞栓を認めた.血液培養よりFusobacterium necrophorumが検出され,抗生剤投与,皮膚切開を行い改善した.咽頭炎から内頸静脈の敗血症性血栓性静脈炎を来し,全身に転移性感染を呈する病態はLemierre症候群と呼ばれる.本症例はその特徴的な経過より同症候群と考えられた.

症例は65歳,女性.発熱,呼吸困難,右下肢痛にて入院となった.入院後,全身状態は急速に悪化し来院6時間後に死亡した.血液培養よりStreptococcus pyogenesが検出され劇症型A群連鎖球菌感染症(toxic shock-like syndrome: TSLS)と診断した.剖検にて,播種性血管内凝固症候群(disseminated intravascular coagulation: DIC)の所見を認め,溶連菌感染がDICの原因と推測された.本例の様なTSLSと考えられる症例では早期診断と治療が救命のためには重要と考えられた.

A novel method for the purification of poly(ADP-ribose) was established. Calf thymus nuclei were incubated with NAD. After pronase digestion, DNA was isolated as a flocculent form by adding 1 M NaCl and 2 volumes of ethanol. Acid-insoluble poly(ADP-ribose), which was synthesized during incubation, was quantitatively recovered in this DNA fraction. Poly(ADP-ribose) was purified as a single peak from a hydroxylapatite column after the nuclease treatment. Preparation of poly(ADP-ribose) thus obtained revealed the characteristics of pure poly(ADP-ribose), including a ratio of A280 to A260 of 0.24.

Clinical and pathological features of eight fatal cases with Sjogren's syndrome (all female, aged 43-61) were studied from the prognostic viewpoint. Chronic active hepatitis, primary biliary cirrhosis, diffuse interstitial pneumonitis and primary pulmonary hypertension were fatal illness in seven of them, and remaining one died of accidental infection. The clinical and pathological findings of these cases suggested that such hepatic or pulmonary diseases might be the manifestations of Sjogren's syndrome. Interstitial nephritis mayalso be a manifestation of the syndrome, possibly with a fatal outcome. Fromthese observations Sjogren's syndrome with liver, lung or kidney involvement may be a possible clinical type with poor prognosis.

Two hybridomas producing monoclonal antibodies to poly(adenosine diphosphate ribose) [poly(ADP-Rib)] were established. One antibody, 10H (IgG3, kappa), bound to most of the poly(ADP-Rib) preparation, which consisted of molecules of various sizes of more than 20 ADP-Rib residues. The binding of this antibody was inhibited by not only poly-(ADP-Rib) but also a monomer unit of poly(ADP-Rib), Ado(P)-Rib-P. The sites protected by antibody 10H were isolated and analyzed by hydrolysis with alkaline phosphomonoesterase and then snake venom phosphodiesterase. The sites contained the same amounts of monomer units and branched portions [Ado(P)-Rib(P)-Rib-P] as the original poly(ADP-Rib) molecules but a lower average number of branched portions per molecule than in the original molecules. The other antibody, 16B (IgM, lambda), reacted with only 50% of the radioactive poly(ADP-Rib), and its binding was not inhibited by a monomer unit. This antibody protected 25% of all the poly(ADP-Rib) molecules from hydrolysis by snake venom phosphodiesterase. The protected sites contained twice as many branched portions per molecule as the original poly(ADP-Rib) molecules. These results show that the two monoclonal antibodies recognize different structures of poly-(ADP-Rib); 10H antibody recognizes the linear structure with ribose-ribose linkages, and 16B antibody may recognize specific structures, including the branched portions of poly-(ADP-Rib).

Poly(ADP-ribose) has been expected to play some regulatory roles in nuclear functions. However, its amount and distribution in vivo have not yet been fully elucidated. An immunohistochemical method should be useful for the study of the natural occurrence of poly(ADP-ribose) at the cellular level. We have developed a highly sensitive immunohistochemical method for detecting poly(ADP-ribose) in conventional histological sections (1, 2).

A portion of the amino acid sequence of human c-Ha-ras protooncogene product p21 is highly homologous with the corresponding region of a family of guanine binding membrane proteins, G-proteins, that are involved in signal transduction (1). Therefore, an analogous function is suggested for ras proteins and G-proteins. Although G-proteins are ADP- ribosylated by cholera toxin (2) or pertussis toxin (3), there has been no data of ADP-ribosylation of c-Ha-ras product by bacterial toxins. On the other hand, both Tsai et al. (4) and our group (5) found ADP-ribosylation of Escherichia coli synthesized c-Ha-ras protein by eukaryotic ADP-ribosyl transferases. Here we identify the amino acid residue which is ADP- ribosylated by a purified hen liver enzyme.

The cytotoxic effects of bleomycin on HeLa cells in culture were enhanced by incubation of the cells with benzamide, a potent inhibitor of poly(ADP-ribose) polymerase, at concentrations at which benzamide alone did not show any cytotoxicity. Benzamide plus bleomycin display enhanced therapeutic effects against Ehrlich ascites tumor cells in vivo. On daily treatment with various doses of bleomycin plus benzamide for 10 days, mice with Ehrlich ascites tumors survived longer than mice on treatment with bleomycin alone.

m-Aminobenzamide and benzamide, which are potent inhibitors of poly (adenosine diphosphate ribose) polymerase and have no antitumor activity themselves, potentiated the antitumor activity of bleomycin against Ehrlich ascites carcinoma in vivo.