Gastric microcirculatory derangement following intermittent electrical stimuli on the main gastric artery produced many gastric ulcers in rat stomach. In this experimental model, the oxyradical generation from two different sources, xanthine-xanthine oxidase system and local neutrophils, were evaluated. Laser doppler flowmetry revealed that sharp fluctuation in mucosal blood flow took place during the repeated stimulation, while systemic blood pressure was unchanged. Thirty minutes after the irritation, the decrease in mucosal ATP contents and the increase of hypoxanthine contents were found. Xanthine oxidase activity in the gastric mucosa also elevated simultaneously. Moreover, both ChL activity of an individual neutrophil and mucosal myeloperoxidase activity were dramatically increased. Superoxide dismutase, chlorpromazine and allopurinol significantly inhibited the formation of gastric ulcers, and attenuated both oxyradical generation of neutrophils and xanthine-xanthine oxidase system. The present result suggests that microvascular derangement can cause the increase in oxidative stress mediated by neutrophils and xanthine oxidase system, which may participate in the formation of acute gastric lesions.
Clinimeal (Eisai, Tokyo) is a semi-digested, low residue diet for tube feeding. We studied the responses of blood sugar, insulin, pancreatic polypeptide (PP), cholecystokinin (CCK) and gastrin using specific RIA methods and exocrine pancreatic secretion and gall bladder contraction using triple lumen tube to intraduodenal ingestion of Clinimeal 100Kcal/hr in eight healthy male adults. After intraduodenal infusion of Clinimeal resulted in a significant rise of blood sugar, serum insulin and plasma CCK from the basal values and returned to the basal level after cessation the infusion. Plasma PP also rose significantly after Clinimeal infusion and sustained the elevation after cessation the infusion. Plasma gastrin, however, did not change. Pancreatic secretion of flow, amylase and bicarbonate output, gall bladder contraction paralleled that of the circulating CCK level. Thus, Clinimeal can significantly stimulate the release of CCK from the upper small intestine and PP from the pancreas, raising the physiological concentration of plasma CCK and PP. This level of CCK can evoke a significant increase in exocrine pancreatic secretion and gall bladder contraction. The sustained elevation of PP may work on an anti-CCK factor.
Activity of ribonuclease (poly Case, poly Uase) in tissues of normal colonorectal mucosa, colonorectal cancer and ulcerative colitis (UC) was measured using synthetic substrates (poly C, poly U). In normal mucosa, poly Case tended to show a higher value in the rectum compared to the colon. In cancer tissue and inflammatory region of UC, poly Case activity decreased, and this activity in the inflammatory region of UC further decreased as the inflammation was intensified. This was considered due to a decrease of non-specific synthesis in cancer tissue and to a decrease of synthesis, acceleration of RNA metabolism and decrease of cells containing poly Case in inflammatory tissue. Poly Uase activity was not significantly different among the three tissues. The above-described changes of poly Case activity strongly suggested that abnormal metabolism of nucleic acids was present in cancer or inflammatory tissue. It was also considered clinicopathologically that poly Case activity may be used as a histochemical indicator of inflammation.
The extractability of neurotensin (NT) from porcine ileal mucosa was evaluated by comparison of eight extraction procedures. Concentrations of NT like immunoreactivities in the mucosa were quantitated and characterized by three sequence-specific radioimmunoassays and gel-chromatography. Tissue levels of intact NT were highest after extraction with boil following homogenization in 2.0M acetic acid (79.1±16.4pmol/g) and lowest after extraction with that in distilled water (6.5±1.4pmol/g). The opposite was the case with levels of N-terminal immunoreactivities (55.2±8.8pmol/g and 105.7±23.8pmol/g, respectively). Recovery studies with addition of synthetic NT1-13 and NT1-8 indicated that these differences could be explained by differences in recovery of intact NT and N-terminal immunoreactive fragments in tissue. In conclusion, this study showed that extraction procedures influence quantitative measurements of NT in ileal mucosa, and that 2.0M acetic acid is preferable when studying intact NT, distilled water is preferable when studying N-terminal fragments, respectively. Therefore, different extraction procedures may be necessary to extract NT like immunoreactive components from tissues.
Morphometric investigations of ballooning hepatocyte and perivenular fibrosis were performed to clarify the pathogenesis of portal hypertension in alcoholic liver disease (ALD). Liver biopsies and measurements of intrahepatic portal vein pressure were performed simultaneously in 8 patient with ALD immediately after abstinence and after liver function test improved. The surface area of hepatocytes and the thickness of fibrous tissue around terminal hepatic vein were analysed morphometrically. The swelling of hepatocytes were more prominent in pericentral zone compared with that in periportal zone. In the patients with ALD with mild fibrosis, the decrease of intrahepatic portal vein pressure were observed in proportion to the improvement of the swelling of hepatocytes after long term abstinence. On the other hand, in the patients with ALD of prominent fibrosis, there was no remarkable change in intrahepatic portal vein pressure in spite of the improvement of swelling of hepatocytes after long term abstinence. The present results suggest that in the patients with ALD in early stage including fatty liver the swelling of hepatocytes is the main factor in the pathogenesis of portal hypertension.
This study is the retrospective analysis of the histological course in 15 cases of HBsAg with anti-HBe. These cases had fluctuating or sustaingly abnormal s-GPT level and these histological prognosis were studied with repeated liver biopsies. In 15 cases, Z value were calculated as follows. Z value=Integral value of GPT more than 50K.U./Integral value of standard GPT (=50K.U.)×Interval time between biopsy (months) Histologically, 6 cases were improved, 4 cases were unchanged, and 5 cases were aggravated. Z value (mean±SD) of improved cases was -54.3±50.8, it of unchanged cases was -15.6±27.6, and it of aggravated cases was 92.8±58.1. Z value of aggravated cases was higher than that of improved and unchanged cases statistically. Therefore, it is concluded that cases with abnormal GPT in anti-HBe positive chronic type B hepatitis should be followed carefully in order to prevent histological aggravation.
Immunoglobulin M antibody to hepatitis B core antigen (IgM anti-HBc) was assayed by radioimmunoassay in serial sera of chronic HBsAg carriers with and without liver diseases. IgM anti-HBc was positive in 525 of 789 sera (66.5%) in cases with histologically proven chronic type B hepatitis, whereas it was positive in only 3 of 98 sera (3.1%) in asymptomatic HBsAg carriers. Cut-off index of IgM anti-HBc was much higher in cases with active liver diseases, and the intrahepatic expression of HBcAg in these cases tended to be located in cytoplasm and membrane of the hepatocyte. During acute exacerbations, the peaks of IgM anti-HBc followed the peaks of sGPT in 40% of the episodes, in 12% the peaks of IgM anti-HBc preceeded those of sGPT, and in 13% both peaks were coincident. Cases with chronic hepatitis persistently positive for IgM anti-HBc were found to develop liver cirrhosis more frequently than those in whom IgM anti-HBc disappeared, and the prognosis of the former were poor as compared with the latter. These results indicate that IgM anti-HBc is closely related to the inflammatory activity and the distribution of intrahepatic HBcAg in chronic type B hepatitis.
The necrotizing effect of the direct injection of anhydrous ethanol on the liver and the subsequent hepatic regeneration were examined in the rabbit. The rather small area (1cm in diameter) of the liver became whitish in color immediately after the ethanol injection. The dilated sinusoids and the slender hepatic cell cords were observed by the histological examination. Then, the white necrotic area enlarged to 4cm in diameter on the fifth day. Massive necrosis of hepatic tissue and formation of thrombi with focal organization of venules were observed. Fibrous tissue appeared around the necrotic lesion 28 days after the injection. The necrotic lesion of the liver induced by the direct ethanol injection healed and disappeared with a small scar in three months. It was concluded that the necrotizing effect of the direct injection of anhydrous ethanol could be attributable to both the direct penetration to the tissue and the disturbance of local circulation due to thrombus formation. These experiments were conducted as a preliminary study for the treatment of hepatic cell carcinoma by the direct injection of ethanol.
The effect of 17α-ethynil estradiol (EE) on biliary cholesterol and bile acids excretion was investigated in the biliary diverted rats. The effect of EE on conversion from free cholesterol of LDL into biliary cholesterol and bile acids was also investigated in the isolated perfused rat liver (IPRL). Wistar male rats were treated with EE for 5 days at 5mg/kg/day. The cholesterol and bile acids of diverted bile for 24 hours were measured with gas-liquid chromatography. [3H] free cholesterol-labeled LDL was infused in to perfusate in IPRL and the radioactivity of biliary cholesterol and bile acids were counted with liquid scintillation counter. In the rats treated with EE, the output and pool size of bile acids, especially of cholic acid, were reduced inspite of no change in the output of cholesterol. And the incorporation of radioactivity of [3H] free cholesterol-labeled LDL into biliary bile acids was reduced. The results sugested that EE reduces newly synthesized cholesterol pool in liver and output of bile acids into bile, and reduced the excretion of bile acids metabolized from free cholesterol of LDL. The lithogenic action of EE may depend on these effects.
An enzymatic method for the quantitative determination of 3-keto bile acid by using of 3-keto-5β-Δ4-dehydrogenase was developed. The data on specificity, reproducibility, sensitivity and accuracy of this assay were satisfactory, and close agreement between the values by this enzymatic method and those determined by gas-liquid chromatography was observed. Consequently this method allowed quantitation of 3-keto bile acids in serum. Then, dehydrocholic acid (DHCA) disappearance curve in serum of healthy subjects and patients with liver cirrhosis after intravenous administration of 1g DHCA was analyzed by using this enzymatic method. In healthy subjects, serum DHCA concentration at 1 minute after loading reached to 364.6μM (in average), and then rapidly reduced to 37.1μM at 20 minutes and 14.6μM at 30 minutes. On the other hand, in patients with liver cirrhosis the clearance of DHCA in serum was significantly delayed, compared with that of healthy subjects.
To determine concentrations of immunoglobulins (Ig) in pancreatic juice, pure pancreatic juice was obtained from patients with acute and chronic pancreatits who underwent endoscopic retrograde intrapancreatic duct canulation. IgG, IgA and IgM were determined by conventional radioimmunoassay, or enzyme immunoassay which was recently developed by us using PINS plates. The values determined by both methods were significantly correlated each other. A main Ig in pancreatic juice from patients with chronic pnacreatitis was demonstrated to be of IgA approximately half of the IgA showed a polymeric form, suggesting strongly the local production. On the other hand, IgG was a predominant Ig in pancreatic juice from patients with acute pancreatitis. As to the IgA molecule composition, the monomeric form was more than the polymeric form. These results suggest that a considerable amount of Ig in the pancreatic juice may be derived from sera of acute pancreatitis patients.
The secretory dynamics of pancreatic juice and protein output in response to a single subcutaneous injection of caerulein (0.1-100μg/kg body weight) were studied in the anesthetized rat. The subcutaneous injection of caerulein at a dose of 0.1μg/kg body weight caused only a slight increase of pancreatic exocrine secretions. Caerulein with doses ranging from 1.25 to 10μg/kg body weight caused a single peaked response of pancreatic juice flow and protein output being maximum during 0-20min after caerulein injection. In contrast, higher doses of caerulein caused two peaks of exocrine pancreatic response, which were seen during 0-20min (first peak) and again during 60-80min (second peak) after the subcutaneous injection of caerulein. Both peaks of exocrine responses showed a biphasic dose-response curve. The first peak of pancreatic exocrine secretions became maximum with 1.25μg/kg body weight of caerulein and then decreased with supramaximal concentrations. On the other hand, second peak of exocrine pancreatic responses increased as the caerulein concentration increased, became maximum with 20μg/kg body weight and then decreased with higher concentrations. When 20μg/kg body weight of caerulein was injected repeatedly 60min after the first caerulein (20μg/kg body weight) injection, the second increase of pancreatic exocrine seretions induced by the first injection was significantly inhibited during 60-100min. The present study revealed that the secretory pattern of exocrine pancreas in response to a single subcutaneous injection of caerulein was quite different (single- or two-peaked response) depending on the injected doses.
Eleven cases of mucinous cystic tumors and eighteen cases of mucous-producing tumors of the pancreas were studied. They were classified into three types: main duct type, branch duct type, and peripheral type. They were analyzed with a special reference to findings of macroscopic and microscopic view, papilla of Vater, pancreatogram and clinical features. The result was that these three types were thought to be the lesions belonging to the same entity. We proposed that these tumors should be named mucin-producing cystic tumor (MCT) of the pancreas.
Span-1 antibody is a murine monoclonal antibody which was produced against a human pancreatic cancer cell line SW 1990. A sensitive sandwich radioimmunoassay was developed using this antibody to evaluate its applicability to the diagnosis of various gastrointestinal disorders including pancreatic cancer. Above 400 units/ml of Span-1 antigen was considered to be positive. Serum Span-1 antigen levels in healthy subjects (n=82) were within 400 units/ml. Positive rating in diseases were pancreatic cancer: 90% (n=79), hepatobiliary cancer: 59% (n=44), gastric cancer: 14% (n=182), colon cancer: 13% (n=50), benign G.I.: 5% (n=86), chronic pancreatitis: 8% (n=25), and acute pancreatitis: 10 (n=10). Levels of serum Span-1 antigen correlated well with treatment and recurrence of disease in a patient with pancreatic cancer. Based on these results Span-1 antigen apparently has good predictive value for pancreatic cancer compared with other gastrointestinal cancers and pancreatitis.