In this study, I purified an insulin-like substance (ILS) in human pancreatic juice by the combined use of affinity chromatography and radioimmunoassay. The amino acid sequence of TLS in the N-terminal region was the same as that of human insulin.
The effect of the enzymes present in the pancreatic juice on the radioimmunoassay procedure was examined. Trypsin, chymotrypsin and amylase showed a marked influences of radioactivity. The addition of enzyme inhibitors did not reduce the pseudo-activity, but the elimination of enzymes in the pancreatic juice by ultrafiltration with the Molecut filter resulted in a lowering of the pseudo-insulin activity. Affinity chromatography on Sepharose 4B coupled with antiporcine insulin was used to capture ILS. Then ILS was eluted by 1M acetic acid from the affinity column which was monitored by the changes of pH and insulin activity in radioimmunoassay.
The amino acid sequences of two components of ILS, separated by HPLC, were Phe-Val and Gly-Ile-Val in respective amino terminal regions. Accordingly, this indicates that ILS obtained from human pancreatic juice was the insulin derived from endocrine secretion of the pancreas.

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The author observed a fact that the incubated splenocytes of the Fischer strain rats with recombinant human interleukin-2 (rIL-2) developed lytic activity to syngeneic natural killer-resistant 9L-glioma cells by a 4-h ⁵¹Cr release assay. The induction of their lymphokine-activated killer (LAK) cell was as follows.
1) When 5 × 10⁶/ml splenocytes were cultured with various concentrations of rIL-2 (50, 125, 250, 500, 1, 000U/ml) for 4 days, the numbers of harvested viable cells increased in responsed to the concentration. The peak was obtained at 500U/ml of rIL-2. And the significant killer activity to 9L-glioma cells was observed at 35-45% (Effector/Target ratio=100/1) in each concentration of rIL-2.
2) When 5×10⁶/ml splenocytes were cultured for 1 to 7 days with 250U/ml rIL-2, the discern-ible lytic activity to 9L-glioma cells was observed on the 1st day culture and with the peak activity on the 4th day.
Moreover, LAK cells prepared from a brain tumor rat showed lytic effect against for the 9L-glioma cells similar to LAK cells from a normal rat.
According to Winn's assay, the growth of tumor was significantly inhibited on the subcutaneous inoculation of 5 × 10⁴ 9L-glioma cells mixed with 5×10⁶ LAK cells.

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In order to estimate the daily intake of calcium, phosphorus, magnesium, iron, zinc, copper, sodium and potassium and to establish their balanced requirement value, two kinds of survey were made of young children at a welfare facility in Osaka prefecture. In survey I, mineral balance tests were performed in order to assess the nutritional status of 18 male and 15 female children (aged from 3y5m to 6y9m) provided with a normal daily diet. In survey II, balance studies of calcium, phosphorus, magnesium, iron, zinc and copper were performed on 15 children (10 male and 5 female aged from 3y10m to 6y5m) receiving two levels of dietary protein which were composed of 2.7 and 2.0 g/kg body weight.
1) The minerals exhibited the positive retention in a daily normal diet. The magnesium, phosphorus, zinc and copper requirements were estimated as 2.6mg/kg, 27.2mg/kg, 197pμg/kg and 36.8 μg/kg body weight respectively.
2) The amount of calcium retention and urinary excretion were not influenced by a decrease in the dietary protein level from 2.7 to 2.0g/kg body weight. However the percentage of urinary calcium excretion showed a higher value, the protein content was increased to the level of a normal daily diet. The amount of urinary excretion of phosphorus increased and retention decreased at the higher protein level compared to the lower protein level (Low protein diet). In regard to the balance of magnesium, urinary excretion was less at the lower level of protein. Intake of copper increased and that of zinc decreased at the lower level of protein compared to the higher protein diet.

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Rat intestinal lactase (EC 3.2.1.23) is separated into two peaks of lactase [I] and [II] on the DEAE-cellulose ion exchange chromatography after papain solubilisation of the brush border membrane. In this study these two lactases were purified individually by ammonium sulfate fractionation and Sepharose 4B, Sephadex G-200, DEAE-cellulose and immunoadsorbent chromatography. The purity of both enzymes was recognized by the single band in polyacrylamide gel electrophoresis and a single precipitation arc in immunoelectrophoresis against antiserum to the papain extract obtained from the brush border membrane of the rat small intestine.
Lactase [I] and [II] had a common antigenity which was shown by the fused arc in immunodiffusion against antisera individually developed from the two purified enzymes. Molecular weight, optimum pH and heat stability were almost identical. However, two enzymes behaved differently on the mobility of electrophoresis, namely Rf 0.244 in [I] and Rf 0.314 in [II].
The total enzyme protein measured by rocket immunoelectrophoresis was at its maximum 10 days after birth (36.5μg/mg brush border protein) and the value in the adult decreased to one twelvth of maximum (3.0μg/mg). Total lactase activity also changed with growth of the rats corresponding to the amount of enzyme protein. The ratio of the two enzyme protein [I]/[II] in the brush border membrane changed gradually from 1.1 in 8 days after birth to 9.8 in adult rats.
These results indicate that there are two kinds of lactase, as examined electrophoretically in the papain extract from the brush border membrane of the rat small intestine. Although these two enzymes are identical in immunoreaction, the ratio of content changed gradually in accordance with the growth, which suggests functional difference between these two enzymes.

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It is well known that Vibrio cholerae non O-1 exist in tropical and subtropical areas. It is considered that they do not exist in Japan. Recently, however, cholerae cases have been found in persons who take fishes and shellfishes. These cases did not have any history of travelling in tropical areas; we think that nowadays Vibrio cholerae widely exist in water.
Vibrio cholerae non O-1 was isolated from sewerage, river water, and sea water in Japan. These evidence suggest chronic water pollution. In order to clarify the environmental ecology of Vibrio cholerae non O-1, we observed the increase and the survival of Vibrio cholerae non O-1 in various conditions, including the following factors; the concentration of NaCI, temperature and nutritional factors. Moreover, we made a comparative study of environmental adaptability between two Vibrio cholerae in different conditions.
The following results were obtained.
1) Vibrio cholerae O-1 and non O-1 could increase neither in the sea nor in river-mouth water at 4°C.
2) Vibrio cholerae non O-1 could increase in the sea and river-mouth water but not in river water at 1O°C. Vibrio cholerae O-1 could not increase in the sea, river-mouth or river water, but could keep the initial number for a long time at 1O°C.
3) Vibrio cholerae non O-1 could increase in river water from the middle and upper reaches, and river-mouth water at room temperature.
4) Vibrio cholerae non O-1 had better adaptability than Vibrio cholerae O-1 to severe conditions for growth; such as high NaCl, low temperature and poor nutritional states.

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It is considered that there exists an equilibrium among the values of serum proteins; alb, y-glb and T.P. The author made an attempt to establish mathematical models upon the data obtained from persons with some abnormalities in liver function tests and from the control ones, respectively. The models obtained had the similar form; Alb=k₁ (T.P.)-k₂ (T.P.) (γ-glb). The calculated constants resembled with each other, notwithstanding the group differences. The equilibrium of the liver damaged group located on low albumin and high y-globulin levels.
The deviation from the control equilibrated state was calculated and this has a statistically significant correlation with the value of zinc sulfate turbidity test (ZTT).
The mathematical plane consisted of the deviation (Q) concerning alb and other values of the serum protein, and ZTT is thought to show the alb and y-glb equilibrium.
The other deviation (D) from the regression line calculated with those two values (Q and ZTT) had a good correlation with serum transaminase values; GOT and GPT, unless GPT exceeds more than 250 Karmen units.

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ENNG was a dministered to 19 6-month-old male beagle dogs at a concentration of 50, 100, 150 or 200pg/ml in drinking water for 3 to 9 months. Total doses of ENNG given were 3.0 to less than 30.0g per dog. In 17 out of the 19 dogs, gastric and/or esophageal cancers were induced and their properties were examined. The gross appearance, location, histological type and metastasis of gastric carcinomas and the incidence of esophageal carcinomas depended on the total dose of ENNG administration.
1) In 6 dogs given totalling 3.0 to 9.9g ENNG, solitary advanced gastric carcinomas of a depressed type were induced mainly in angulus ventriculi, most of which were poorly differentiated adenocarcinomas and partially composed of signet-ring cell carcinomas. All of these dogs showed lymph nodes metastasis. No esophageal cancers developed in any of these dogs.
2) In 7 dogs given 10.0 to 14.9g ENNG, protruded type carcinomas were frequently observed in the antrum. In nearly all these tumors well differentiated adenocarcinomas were observed, in addition to poorly differentiated and signet-ring cell carcinomas. Lymph nodes metastasis was found in one third of these dogs. Esophageal squamous cell carcinomas also developed in one third of these dogs.
3) In 4 dogs given 15.0 to 30.Og ENNG, depressed type carcinomas were often found in the pyloric gland region. In many of these tumors well differentiated adenocarcinoma was also partially observed along with poorly differentiated and signetring cell carcinomas. Lymph nodes metastasis was observed in 2 out of 4 dogs. Esophageal carcinomas were also induced in three fourths of the dogs under this condition.
4) Fifteen gastric polyps developed in 7 dogs associated with gastric carcinomas. The number of polyps tended to increase, with the increase of the total dose of ENNG. Poorly differentiated adenocarcinoma was partially observed in 7 of the 15 polyps.
The above results indicate that by setting up the total dose of ENNG in drinking water, experimental canine gastric and esophageal carcinomas, with the morphological features which satisfy experimental purposes, could be induced.

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The mechanism of iron uptake in isolated hepatocytes of Wistar strain female rats was investigated by primary monolayer culture for 24h at 37°C. The results were as follows:
1) Iron uptake from diferric transferrin (diFeTf) showed different processes dependent on the ligand concentrations. Namely, the receptor-mediated transport was observed in a lower concentration of diFeTf from 0 to 0.2μmol/l, while another passive process was suggested in a higher concentration of the ligand from 15 to 32μmol/l.
2) About 90% of ⁵⁹Fe from transferrin was found in a whole membrane fraction of hepatocytes and the remainder was in cytosol. ⁵⁹Fe and ¹²⁵I activities from radiolabelled diFeTf incorporated into the whole membrane fraction were localized mainly in plasma membrane and lysosome fractions, which were identified by Percoll density gradient centrifugation and simultaneous marker enzyme analyses.
3) Receptor assays for diFeTf and diferric asialotransferrin were performed. Ka and the binding number were 4.9×10⁷M^-1 and 3.85×10⁵ (high affinity transferrin receptor), 2.2×10⁶M^-1 and 2.81×10⁶ (low affinity transferrin receptor), 8.0×10⁷M^-1 and 5.22×10⁵ (asialotransferrin receptor). The competitive bindings between diFeTf and diferric asialotransferrin to each receptor were recognized. These phenomena suggested that both receptors had a common binding for each ligand.
4) Iron uptake into hepatocytes from ferrous citrate increased linearly according to increasing ferrous citrate concentration from 0.3 to 120μmol/l by a non-specific process. The total amount of iron incorporated into hepatocytes from ferrous citrate was about 5 fold higher than the uptake of the same concentration of transferrin.
5) The percentage of ferritin 59Fe in whole activities from cytosol and heat extract of membrane fraction was 5 fold higher in diFeTf uptake than that in ferrous citrate, which was detected by immunoprecipitate against anti rat liver ferritin serum.
6) Iron uptake from diFeTf was faster in the presence of ferrous citrate in the incubating medium than in diFeTf alone.

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