Yoshio Hatta

Gas chromatography of dimethylalkylsilyl ether derivatives of bile acid methyl esters

The gas chromatographic separation of the DMAS ether derivatives of ten bile acid methyl esters, namely DMES, DMnPS and DMiPS ethers, have been studied by use of an open tubular glass capillary column, coated with SE-30. The DMAS ether derivatives were eluted in sequence according to the number of hydroxyl groups except of 12-KCDCA, and separated. This contrasts with the poorly resolved gas chromatographic peaks produced by the TMS ether derivatives. The DMES ether derivatives were resolved and baseline separation in 30 min, but those of HCA and 12-KCDCA had similar methylene unit values.

Inhibitory effects of the cholecystokinin antagonist loxiglumide on pancreatic exocrine secretion and pancreatic growth in conscious rats

SummaryThe effects of cholecystokinin (CCK) receptor antagonist Loxiglumide (CR 1505) on pancreatic exocrine secretion and growth stimulated by chronic bile-pancreatic juice diversion to the ileum were studied in conscious rats. Pancreatic secretion was measured each day at 0900 h for 7 d. Pancreatic flow and protein output were significantly increased 24 h after bile-pancreatic juice diversion. Protein output increased each successive day, reaching maximal values of 3.6-fold above basal by the 6th and 7th d of chronic bilepancreatic juice diversion. Fluid output reached maximal values of approx 3.5-fold above basal by the 3rd d of chronic bile-pancreatic juice diversion. Plasma CCK increased threefold above basal levels after 24 h of bile-pancreatic juice diversion and remained three- to fourfold above basal. Intragastric bolus infusion of CR 1505 (50 mg/kg) on the 7th d of chronic bile-pancreatic juice diversion inhibited pancreatic protein and fluid secretion by 80 and 75%, respectively, 60 min after administration and by 52 and 71%, respectively, 5 h later. Pancreatic wet wt after 7 d of chronic bile-pancreatic juice diversion was significantly increased by 56%, and this was completely suppressed by 50 mg/kg of CR 1505 given intragastrically every 12 h. These rests indicate that the rat with chronic bile-pancreatic juice diversion is a useful model to examine both potency and duration of the action of CCK receptor antagonists and show that CR 1505 inhibits pancreatic exocrine secretion and growth induced by endogenous CCK.

Hyperamylasemia and S-type isozyme dominance in liver cirrhosis

SummaryTo investigate the mechanisms of serum amylase abnormalities in liver disease, we determined serum amylase levels, S-type isozyme proportion, clinical symptoms, and laboratory data in 38 cases of histologically confirmed liver cirrhosis and 19 controls. Of the 12 patients who were hyperamylasemic (12/38, 32%), 5 showed S-type isozyme dominance (5/12, 42%), whereas in the 26 normoamylasemic cirrhosis patients, only 5 were S-type isozyme dominant (5/26, 19%). Isozyme percentages were significantly higher (P<0.01) in the dominant-S-type cases than in the controls, and S-type dominance was found more frequently in the hyperamylasemic than in the normoamylasemic cirrhosis cases. Only ascites and esophageal varices were observed more frequently as clinical symptoms in the dominant-S-type cases. Our results suggest that amylase is not produced in the human liver, but that the decreased clearance rate of amylase, especially the S-type isozyme, may be a cause of hyperamylasemia and S-type isozyme dominance in cirrhosis.

Changes in immunoreactive insulin, C-peptide immunoreactivity, and immunoreactive glucagon in acute viral hepatitis

Insulin and glucagon are thought to play important roles as hepatotrophic factors in acute viral hepatitis (AVH); however, few reports have investigated the responses and relationships of each of these hormones to liver damage in detail. We studied insulin and glucagon responses during the acute and recovery phases of AVH. We performed a glucose tolerance test (GTT) and an insulin sensitivity test (IST) in each phase in 11 patients with AVH. In 8 additional patients in the acute phase (totaln=19), were compared immunoreactive insulin (IRI) and C-peptide immunoreactivity (CPR) levels with transaminase levels. In the acute phase, IRI concentrations were normal from fasting to 60 min, despite an increased CPR level. In the recovery phase, IRI and CPR levels increased significantly. Immunoreactive glucagon levels in both phases did not differ significantly from those in controls. During the IST, the insulin sensitivity index in both phases was significantly lower than that in the controls. Fasting IRI and ΣIRI showed significant negative correlations with transaminase levels. We found enhanced insulin secretion and a decrease in plasma insulin in the acute phase of AVH. The discrepancy between IRI and CPR responses in the acute phase suggests an increase in the degradation or consumption of insulin in the liver.

Symposium (III): Clinical aspects of the duodenal diverticula

bu t not remaks, 12 months after biopsy done, adenocarc inoma tubu ia re was indicated, it is I I a type early cancer. Next we showed a few cases of confusable benign pro t ruding type lesions in the prepyloric portion. Case 3 is a gastritis verccosa, case 4 was a benign ulcer of near the rings, bu t looks like Bor rman I I type cancer, as the h igh land walls, case 5 was abe r r an t pancreas, case 6 was benign mul t ip le cysts in the submucosa and last case was adul t pylorusring muscle hyper t rophy. W e are conclusive in this studies, it is more i m p o r t a n t biopsy study to diagnosis or different ia t ion I I a and inf lammatory changes of prepyloric portion, and so necesary to examine by duodenoscope to observe in details,

Clinical evaluation of per-rectal portal scintigraphy in patients with liver cirrhosis.

99mTcを用いた経直腸門脈scintigraphyを肝硬変51例に施行し,肝血流と門脈shunt形成につき臨床的に検討した. RI activityの1分値と15分値のカーブの解析より,肝先行型(I型)8%,肝心同時出現型(II型)43%,心先行型(III型)49%に分類された.臨床所見との比較では,食道静脈瘤と脾腫はIII型に著明であった(p<0.01).肝細胞障害を示す諸検査とでは,各型に有意差を認めなかつた.また, ICG(R15)検査では各型間に有意差を認めた(p<0.05).肝機能に異常を認めない特発性門脈圧亢進症ではIII型を示した.以上,非侵襲的検査としての経直腸門脈scintigraphyはshunt形成のみならず,肝血流状態を反映し,かつ肝硬変の進行度をある程度予測するのに役立つと思われた.

Proceedings of the 19th autumn meeting from October 18 to 20, 1977-Nara, Japan

The first step in the formation of bile acids is 7uhydroxylation of cholesterol. Subsequently 7ahydroxycholest-4-en-3-one is formed, which is a common precursor of the two primary bile acids, cholic acid and chenodeoxycholic acid. 5,BCholestane-3~t , 7a -d io l , a p recursor of chenodeoxycholic acid is formed from the A4-3-keto intermediate by a pathway which involves the reduction of the keto group and the saturation of the double bond but not a hydroxylation at C-12, while a pathway involving the 12a-hydroxylation of the A4-3-keto intermediate leads to the formation of 5flcholestane-3~, 7a, 12a-triol, which is a precursor of cholic acid. The C2~-bile alcohol intermediates , 5flcholestane-3a, 7t~-diol and 5,g-cholestane-3tz, 7t~, 1 2 ~ t r i o l , a r e t h e n t r a n s f o r m e d i n t o chenodeoxycholic acid and cholic acid by the degradation of the side chain which entails an moxidation followed by fl-oxidation. Chenodeoxycholic acid is also formed by a number of different pathways differing only with respect to the stage of nuclear transformation at which oxidation of the side chain is initiated. A number of cholestanepolyols such as 5flcholestane-3ct, 70t, 12ct, 25-tetrol are accumulated in the bile and feces of patients with cerebrotendinous xanthomatosis. No 26-hydroxylated bile alcohols are accumulated. The results suggest that there exists an alternative pathway of cholic acid biosynthesis involving the 25-hydroxylated bile alcohol as an intermediate. (2) Analytical methods of bile acids in biological materials

Metabolism of ceruloplasmin in liver disease

Iron binding tissue protein, ferritin is known to have diffeernt electro-phoretie mobiIities according to the tissues of which ferritin was isolated. In the present report, ferritin prepared by a mild and less cumbersome method of chromatographic isolation was studied by isoelectric fractionation and disc electrophoresis to assess the differences of ferritin molecules obtained from different species and from different organs. The experimental conditions for electrofocusing were the same as described by Vesterberg et al., except for the use of prefocused carrier ampholytes with much narrower pH ranges to facilitate more accurate determination ofpIs of the components under investigation. The electrophoretic mobilities of the f~rritin samples were the fastest in horse splenic ferritin and slightly delayed in order of human liver ferritin, rat liver ferritin, dog liver ferritin and dog splenic ferritin. Each preparation consisted of mainly monomeric form and contained lesser quantities of dimer and polymer types. The ranges of pls determined by electrofocusing were; 3 main fractions in the pH range of 4.2 to 4.6 for rat hepatic and splenic ferritin, in the pH range of 5.0 to 5.4 for dog hepatic ferritin, in the pH 5.0 to 5.7 for human hepatic and splenic ferritin, the latter was found to have more neutral pIs than the former. Changes of iron to protein ratios following iron loadig and artificial bleeding treatment were studied in rats and only bleeding caused a considerable lowering of the ratio in the ferritin fractions having higher pl values. Relationship between iron mobilization and ferritin microheterogeneity was discussed.

Clinical significance of quantitative study of carboxypeptidase in pancreatic juice

SummaryThe determination of the volume, amylase content and bicarbonate concentration in pancreatic juice after Pancreozymin-Secretin stimulation had been considered to be valuable for the diagnosis of pancreatic diseases. However, since these values were not always coincidental, the diagnosis of pancreatic diseases were difficult in many cases. To have a more difinite criteria of Pancreozymin-Secretin test, the content of carboxypeptidase A and B in the juice was compared with other results and the secretory mechanism of this enzyme were investigated by the method of radioactive zinc.As the results, it has been provided that the determination of carboxypeptidase A and B was also as useful as the measurement of the volume, amylase out put and maximum concentration of bicarbonate for more distinct diagnosis qualitatively and quantitatively of pancreatic disease on Pancreozymin-Secretin test.The maximal radioactivity was obtained in the pancreatic juice between 3 to 6 hours after intravenous injection of Zn65 in normal rats. The radioactivity and the carboxypeptidase activity in the juice was completely paralleled by sephadex and cellulose column Chromatographie analysis.Thus, it was assuemed that Zn65 was excreted as a component of that enzyme protein moiety and time needed for synthesis and excretion of that enzyme might be between 3 to 6 hours.

Study on gastrin (I)

Some exper iments were performed to invest igate the physiological and clinical significance of Gastrin. 1) Antibody production. The C-terminal tetrapeptide of gas t r in was repeatedly injected into the foot pad of rabbi t for 5 to 6 t imes. 2) The rabb i t serum incubated wi th 3H-labeled tetrapeptide was analysed by Sephadex G-25 column chromatography. The antigen-antibody bound protein peak was obtained and the immunoassay method was fu r the r studied. 3) The 3H-tetrapeptide or 12H-hexapeptide disappeared f rom the p lasma almost lineally for 30 rain. a f te r injection. From the resul t s of the radioact ivi ty of the organs, it is assumed that these peptides might be taken up by the gas t r in excretory parietal cells in relat ively early t ime af ter being injected. It was not assured whether the endogenous and exogenous gas t r in was metabolized in the same way as the resul ts in th is experiments , scince only the C-terminal peptide were used instead of the whole gas t r in molecule.