Chemical changes in the blood induced by maltitol were compared with those induced by glucose in both healthy people and patients with several disorders including diabetes mellitus.
Blood glucose levels of healthy subjects were determined after the administration of glucose (12.5g, 25g, 50g) and maltitol (50g). Based on the glucose absorptin curve, 38% of the maltitol orally administered was absorbed through the intestinal tract, but the absorption of maltitol was more delayed than that of glucose.
After administration of maltitol to the groups with diabetes and the other diseases with impaired glucose tolerance, the blood glucose level was higher than the level in healthy people. Elevation of blood glucose level by maltitol was 25 to 50% of the elevation by glucose, and there was a correlation between the elevation of blood glucose level by glucose and by maltitol.
Although elevation of blood IRI was parallel to the doses of glucose in healthy people, the level of IRI following 50g maltitol administration was significantly lower than that following 12.5g glucose. The elevation of IRI caused by 50g glucose was lower than that caused by 50g maltitol in various diseases (normal GTT type, borderline type and diabetic type of cases more than 60 years old).
The peak blood glucose level after administration of a mixture of 50g glucose and 50g maltitol was lower than that of 50g glucose alone. This result was discussed by the author.
The adverse reaction after oral administration of maltitol was diarrhea, which was frequently observed in healthy people and the patients with the borderline pattern of glucose tolerance test. However, the frequency of diarrhea was very low in the patients with diabetes and the other diseases with impaired glucose tolerance.

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The insulin release induced by arginine was studied in normal subjects and hyperthyroid patients. Arginine hydrochloride (0.5g/kg of body weight) dissolved in 200 ml of physiological saline was administered to normal subjects and hyperthyroid patients by intravenous infusion in a cubital vein over 30 minutes after overnight fasting. Blood samples were taken before and 2, 4, 6, 8, 10, 15, 30, 45 and 60 minutes after the start of the infusion. Plasma insulin was determined by double antibody radioimmunoassay and blood glucose by autoanalyzer. Biphasic insulin release by arginine infusion was observed in normal subjects. The AIRI of the first peak at 4 minutes was 23.6+17.4μU/ml, and the second peak at 30 minutes was 40.9+10.6μU/ml. Monophasic insulin release by arginine infusion was observed in hyperthyroid patients. The AIRI of the first peak at 2 minutes was 13.6+2.7μU/ml, but the second peak was very low. The correlation coefficient between AIRI and ABS was-0.08 (not significant) in the first phase of insulin release and 0.72 (p<0.01) in the second phase of insulin release. This suggests that the second phase of insulin release is influenced by the elevation of blood glucose by arginine infusion. The correlation coefficient between AIRI and RSU was 0.22 (not significant) in the first phase of insulin release and-0.71 (p<0.01) in the second phase of insulin release. We observed a difference in character between the first and the second phases of insulin release at correlation between AIRI and ABS, AIRI and RSU and concluded that the first and the second phases of insulin release by arginine infusion seem to occur from the different mechanisms of insulin release.

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A modification of the dextran coated charcoal method for immunoassay of insulin is described which offers certain advantages in use: a plain charcoal method for insulin.
First, the charcoal adsorption rate of insulin and the insulin antibody complex were studied with dextran coated and uncoated charcoal in various concentrations. No apparent difference in adsorption was recognized between dextran coated and uncoated charcoal. Changes of the charcoal adsorption rate were also studied in various concentrations of serum (6% to 33%) in assay mixtures, and a quite similar tendency of decrement in adsorption rate was observed both in dextran coated and in uncoated charcoal.
The adsorption curves related to the protein concentration suggesting that the proteins are in competition with the free insulin as has been emphasized by other investigators. The most efficient concentration of plain charcoal was found to be 0.17% of the final concentration in the presence of 0.1m/ of serum. It should be noted that the serum content of all tubes must be maintained constantly. Practically, the plain charcoal method of insulin radioimmunoassay was carried out and proved to be satisfactory sensitive, reproducibile and relatively inexpensive. It was also simple to perform.

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Changes in the small blood vessels of the nailbed were examined by vital capillaromicroscopy in 17 diabetic children. Abnormal morphological changes such as dilatations, wavings, tortuosities, branchings, nodular apical elongations, tanglings, granularities and so-called fine capillaries were observed in each photographic picture. Points were given according to their severity, and then they were compared with the findings in adults at the onset of diabetes. The nodular apical elongations were increased significantly in diabetic children compared with those of nondiabetic children. The findings of granularities and fine capillaries seemed to indicate that they were changes due to aging rather than to diabetic metabolic abnormalities.

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The pancreatic islets of rabbits treated with l-asparaginase (1000U/kg) or with 1-asparaginase and cortisone (10mg/day) simultaneously, were examined histologically and electron microscopically.
1) It was proved by chromium-alum-hematoxylin staining treatment that a single dose of l-asparaginase resulted in different degrees of degranulation of B-cells.
In electron microscopic examination, a remarkable decrease of B-granules was observed. Almost all of the presecretory granules and limiting membranes were lined up along the marginal zone of the cytoplasm. The Golgi apparatus was enlarged enormously and dilation of their lamelle was observed. Development of endoplasmic reticulum was prominent in several cells. Vacuoles and lysosome-like bodies were observed in the cytoplasm of the B-cells.
2) Degranulation of the B-cells was found to be more prominent in the group treated with a combination of 1-asparaginase and cortisone than in the group treated with 1-asparaginase alone. A few B-granules were localized along the margin of the B-cell membrane. Nuclei of the B-cells appeared to be intact and no severe destruction of the B-cells was found. In electron microscopic examination, the decrease of B-granules was remarkable and residual secretory granules were located adjacent to the plasma membrane. Mitochondria were swollen to some extent and the cristae were found to be irregular in a lattice work, or they showed a stellate appearance. The Golgi appearatus was decreased significantly in size and number. Increases of free ribosomes were found. They were released from the endoplasmic reticulum and were scattered in the cytoplasm of B-cells. Filament structures were increased prominently in the cytoplasm.

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It is generally accepted that the insulinogenic index is low in diabetes mellitus and rather high in secondary diabetes. Some secondary diabetics, however, show a low insulinogenic index. An investigation was carried out to determine whether this low insulin response of non diabetics to glucose is essentially identical with that of diabetes mellitus. The insulin response to a 50g glucose tolerance test (GTT) and a 50g glucose tolerance test combined with 2.5mg of oral glibenclamide (G1-GTT) was determined in 46 subjects, using JIRIMBS, EBS and EIRI as an indicator of insulin secretion. They were classified into four groups according to the GTT curves: normal, diabetic, secondary diabetic and borderline group.
In the diabetic group (16 cases), the IRI/BS (30 min) indices were below 0.4 both in GTT and GI-GTT with one exception, even though they were higher in Gl-GTT. In the secondary diabetic group (17 cases), the IR/BS indices were below 0.4 in six cases in GTT. while the others revealed normal insulin responses. These six cases showed IRI/BS indices above 0.8 in Gl-GTT. Five of six cases had low ΣIRI and high ΣBS in GTT and were hardly distinguishable from the diabetic group. In Gl-GTT, ΣIRI increased significantly in five non diabetic cases, but did not increase in diabetics. Borderline cases were similar to secondary diabetes rather than to diabetes mellitus in insulin response to Gl-GTT. It was concluded that some secondary diabetics and borderline cases show low insulin response to GTT and that the low insulin response of nondiabetics can be augmented in the Gl-GTT, but in diabetes it cannot. Gl-GTT might be a valuable method for differentiating insulin low responders.

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Pancreatic glucagon (IRG), insulin (IRI) and blood sugar (BS) were measured in 12 non-obese controls and in 58 untreated, recently diagnosed diabetic patients during an oral glucose tolerance test. The diabetic groups were classified as “mild” ones (fasting blood sugar, FBS, below 140mg/dl), “moderate”(140<FBS<200mg/dl) or “severe”(FBS above 200mg/dl). The glucagon values reported were obtained by using an antibody (30K; Unger) to pancreatic glucagon which showed negligible cross-reaction with gut extracts. After glucose load, specimens were drawn at 30, 60, 90 and 120 minutes. In the controls IRG decreased successively and the fasting IRG, 98±5 pg/m/, declined to 63±5 and 60±6 pg/m/ at 90 and 120 minutes, respectively (p<0.01). In “mild” diabetics the IRG also decreased but not significantly. In “moderate” and “severe” diabetics IRG didn't decrease significantly and it tended to elevate paradoxically despite prolonged hyperglycemia. The reasons why the IRG was elevated paradoxically, after an oral glucose load in “moderate” and “severe” diabetics were not known, although the participation of gut glucagon could be excluded.
In order to elucidate the responsiveness of pancreatic alpha cells to glucose, summations of the changes of IRG or BS at each minutes from the the fasting level after glucose load were caluculated. When IRG or BS became smaller than the fasting level, the difference in value was expressed as a negative sign. The ratio -ΣIRG/ΣBS was 1.96±0.57, 0.11±0.05, -0.002 ± 0.06 and -0.09±0.07 (mean +SEM), in the controls, “mild”, “moderate” and “severe” diabetics, respectively. The data indicate that the ratio of the degree of suppression of the pancreatic glucagon release to the glycemic stimulus was distinctly less in diabetics than in normal controls, and that it became still less with the exacerbation of the diabetes mellitus.
From these findings, we speculate that the ratio -ΣIRG/ΣBS after OGTT will be useful in the differential diagnosis of glucose intolerance as a new tool, together with the insulinogenic index at 30 minutes.

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Some 200, 000 death certificates from 1967 to 1971 were reviewed of which 3, 261 mentioned diabetes as a cause of death. Of these 1, 883 recorded diabetes as the underlying cause and 1, 378 listed diabetes as a contributory cause. Both were analyzed and compared with similar studies in the United States (US Public Health Service 1955) and (Tokuhata et al. Pennsylvania 1968-69) in order to determine the differences in the causes of death between the two countries. Diabetes was the underlying cause in 57.7% of the cases in Japan compared to 40.8% in the U.S.A. and to 25.9% in Pennsylvania. Of contributory conditions (2.5 average in Japan and 3.13 average in the USA) in cases with diabetes as the underlying cause, 72.4% were diseases of the cardiovascular system in the U.S.A compared to 54.7% in Osaka. Arteriosclerotic heart disease and general arteriosclerosis were predominant in the U.S.A but hypertensive disease and nephropathy were more frequent in Osaka. In analyzing the distribution of underlying causes of death where diabetes was a contributory condition, a statistically significant excess of death was observed in diabetics from nephritis and nephrosis, cirrhosis of the liver, hypertensive disease and tuberculosis over the expected number. The most frequent underlying cause of death was arteriosclerotic heart disease in the U.S.A and cerebrovascular disease in Osaka. Tuberculosis, malignant neoplasm of the stomach and cirrhosis of the liver prevailed in Japanese diabetics. A marked difference in the causes of death was found between the two countries which seemed to be largely dependent on the characteristics of the population in each country.

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Although the best-known effect of glucagon is an increase in blood glucose concentration, early preparations of glucagon were contaminated by a considerable amounts of insulin. This led to the reinvestigation of the degree and duration of the hyperglycemic response using a recent glucagon preparation which contained very little insulin (5mU/mg), since it is useful to know the intensity and duration of the response. On the other hand, glucagon has been shown to have effects on lipid metabolism and adipose tissue, although little is known about the influence of glucagon on the cholesterol and triglyceride concentrations in blood. Since the etiological significance of cholesterol and triglyceride in arteriosclerosis, diabetes mellitus and related diseases has been discussed, it is interesting to know the effect of this hormone on these lipids.
Glucagon (Lilly), 1mg dissolved in 20ml of physiological saline, was injected intravenously into 16 normal nonobese controls (N) and 32 untreated nonobese diabetics. The diabetics were classified as D₁ (11 cases, fasting blood glucose under 129 mg%), D₂ (12 cases, fasting blood glucose 130-199mg%) and D₃ (12 cases, fasting blood glucose>200 mg%).
1. Peak values of blood glucose concentration were obtained after 15-30 minutes in N, after 30 minutes D₁ and D₂, after 45 minutes in D₃.
2. The mean maximum increment in blood glucose was 44±4.3 mg/dl (mean +SEM) in N, 57±6.6 mg/dl in D₁, 80±5.6 mg/dl in D₂ and 78±12.3 mg/dl in D₃.
3. Blood glucose returned to basal level 60 minutes after the injection in N, 90 minutes after the injection in D₁, 90-120 minutes in D₂ and 120 minutes after the injection in D₃. These results suggest that the glucagon tolerance test reflects fairly well the severity of diabetes.
4. Blood cholesterol concentration showed no change or showed a very slight and transient decrease (0.9%-1.37%) after glucagon injection, but glucagon induced a meaningful decrease (p<0.005) in triglyceride concentration both in controls and diabetics. Mean fasting levels of triglyceride are 106±15.5 mg/dl in N, 95±12.2 mg/dl in D₁, 136±16.0 mg/l in D₂ and 153±26.6 mg/dl in D₃. The mean maximum decrease are 16.8% in N, 17.7% in D₂ and 24% in D₃120min. after injection. In D₁, however, the change was shown to be not constant. The mean maximum change is only -5.3% at 60min.
The effect of glucagon on plasma lipid was discussed.

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The correlation between 12 variables retinal findings and 9 variables clinical laboratory findings in 223 diabetics was studied by multivariate analysis such as correlation analysis, factor analysis (varimax method) and discriminant function. Hammer-Smith Hospital standards set by Oakley et al. were adopted for the statistical analysis of retinal findings and claculations for multivariate analysis were carried out with a model NE AC 2200 computer.
Factor analysis revealed that diabetic retinopathy was divided into “simple retinopathy” composed of deep haemorrhages and hard exudates, and “malignant retinopathy” composed of new vessel formations and fibrous proliferation. Superficial haemorrhages were closely related to the latter.
i) Stages of diabetic retinopathy by Scott's classification were closely related to systolic blood pressure, duration of diabetes mellitus and fasting blood sugar.
ii) Deep haemorrgages were closely related to systolic blood pressure, fasting blood sugar and urinary protein; superficial haemorrhages to fasting blood sugar, BUN and urinary protein; hard exudates to serum cholesterol; and new vessel formations to fasting blood sugar, serm cholesterol, systolic blood pressure and urinary protein.
iii) First component of factor analysis representing malignant retinopathy was closely related to fasting blood sugar and urinary protein. The second representing simple retinopathy was closely related to serum cholesterol, fasting blood sugar and systolic blood pressure.
Discriminant functions for predicting malignant retinopathy applying retinal and clinical laboratory findings were composed of a linear combinating formula. Probability by this function was 97.6%. Multivariate analysis is also useful in determing the prognoses of diabetics with retinopathies.

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Pancreatic scintiphotography was performed in 108 cases of patients with diabetes mellitus. Scintiphotos were taken at 30 min. after intravenous injection of approximately 200pCi of ⁷⁵Se-selenomethionine using a Toshiba gamma camera. The relationship between the degree of pancreatic uptake of ⁷⁵Se-selenomethionine and the types and duration of diabetes, vascular complications and the average range of fasting blood sugar levels were studied. In some cases, pancreatic scintiphotos were taken at 10, 30 and 50 min. after injection of ⁷⁵Se-selenomethionine, and the degrees of the pancreatic uptake were compared on each time course. Only two out of 24 cases of insulin-dependent diabetics showed normal pancreatic scintiphotos. On the other hand, two out of 47 cases of mild diabetics treated with diet alone showed no uptake in pancreatic scintiphotos. There was a tendency toward abnormal pancreatic scintiphotos in chronic diabetics. Especially, of the 15 cases who had diabetes for more than eleven years, only one case showed a normal pancreatic scintiphoto. Abnormal pancreatic scintiphotos were found more frequently in the group of poorly controlled diabetics than in the group of well controlled diabetics. In cases showing normal pancreatic scintiphotos, diabetic retinopathy was less frequently found. Out of 36 cases which had sequential pancreatic scintiphotos, hypertension and/or arteriosclerosis were found more frequently in the 20 cases which showed a delay in reaching a plateau of the activity. However, the uptake in sequential pancreatic scintiphotos showed no definite correlation between diabetic retinopathy and other diabetic conditions.

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The authors studied the binding of insulin to rabbit erythrocytes, as a mode case in the hope of characterizing the physiologic role of the binding of insulin to receptor in both normal adults and patients. Specific binding sites for insulin were detected in rabbit erythrocytes. The characteristics of the binding were similar to those observed in other target tissues. The specific binding of ¹²⁵ I-labeled insulin was competitively inhibited by a small amount of unlabeled insulin and was completely inhibited by 1, 000ng/ml of unlabeled insulin. Glucagon, however, had no effect on the insulin binding to fat cells or liver membranes nor had it any effect on the binding of insulin to rabbit erythrocytes. Scatchard analysis of this binding reaction indicated two different binding sites with K_aff=3.2×10⁸/M, K_diss=3.1×10^-9M; K_aff =1.4×10⁸/M, K_diss=7.1×10^-9M respectively, and the binding capacities of each site were estimated at 0.011ng/4×10⁸ cells and 0.138ng/4×10⁸ cells. The binding of ¹²⁵I-insulin to rabbit erythrocytes was a saturable function of the insulin concentration and was a linear function of cell concentration. The pH optimum for the reaction was 7.4. At 0°C, the amount of insulin binding increased continuously under the reaction and this binding reaction reached a steady state after 10 to 15hr. On the other hand, the specific binding of insulin at higher temperatures showed maximal amounts after 20 to 30min. and subsequently fell off at later time points.

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