The effects of dietary lipid and daily food-uptake on hepatic HMG-CoA reductase (β-hydroxy-β-methylglutaryl CoA reductase) activity and serum cholesterol level of the normal and diabetic rats were studied.
The animals were maintained ad lib on either carbohydrate diet (dextrose 75%, casein 20% and vitamins et salts 5%) or fat diet (dextrose 30%, casein 20%, vitamins et salts 5%, corn oil 20% and indigestible fibres 25%) for five days from 8 o'clock in the morning till 6 o'clock in the evening. Daily food consumption per animal under these conditions were nearly 60 calorie and no significant differences were observed. The other groups of normal and diabetic animals forcedly fed one of the two kinds of the diet 76 calories daily for four days, were also studied. The measurements of the enzyme activity were done at noon and midnight. The serum samples were obtained at noon and used for cholesterol analysis.
The diurnal variations of the enzyme activity were shown on the all groups observed. The lipid feeding markedly stimulated the HMG-CoA reductase activity of both normal and diabetic rats. Increased consumption of the diets also stimulated the enzyme activity in most of normal and diabetic animals. However, these stimulatory effects on the reductase activities by the different dietary states were neither accompanied by increase nor decrease of serum cholesterol level.
These discrepancies between the hepatic cholesterogenesis and serum cholesterol level suggest that the liver has rather limited contribution to the whole cholesterogenesis or that the breakdown of serum cholesterol is also stimulated coordinately with it's hepatic production by these dietary manipulations. The possible mechanisms of the diurnal variations of the activity and the indUction of hepatic HMG-CoA reductase by dietary lipid or increased food uptake were discussed.

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A-component in the crystalline insulin was one of impure substances of higher molecular weight contaminating the conventional insulin solution sold before June, 1975. The blood samples used in this study had been collected before May, 1975.
The presence of specific anti-a-component antibodies was examined in sera of 14 patients who had circulating anti-insulin antibodies possibly produced by autoimmunity, six of them showed insulin autoimmune syndrome with spontaneous hypoglycemia and the remaining 8 patients had only the autoimmune antibodies without hypoglycemic attacks. Sera of 14 diabetics treated with conventional insulin for more than one year were also examined.
Percentages of ¹²⁵I-a-component bound with sera of all the patients of both groups were higher than normal. However, after preincubation of the sera with cold Monocomponent Insulin, the percentages of ¹²⁵I-a-component bound with sera of all the patients with insulin autoimmunity entered in the normal range with one exceptional case.
On the contrary, the percentages in patients treated with insulin were still higher than normal.
The serum of one diabetic patient who had been treated with Monocomponent Insulin and developed anti-insulin antibody showed the same pattern of binding with 125I-a-component as the sera of the autoimmune group.
In conclusion, the specific antibody to a-component was found only in sera of the patients treated with conventional insulin. And the detection of anti-a-component antibodies might be one of important methods to differenciate the insulin autoimmune syndrome from factitious hypoglycemia appeared before June, 1975.

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Sodium ion is considered to be an essential factor for insulin release from the islets of Langerhans in addition to calcium ion. However, inter-relationship between these two ions is still obscure. A commonly used anticonvulsant, diphenylhydantoin (DPH) has been reported to inhibit insulin release in vivo and in vitro, but its precise mechanism has not been clarified. Recently, DPH was shown to reduce Ca uptake in addition to sodium uptake in nerve cells. This paper presents the evidences that alterations of sodium and calcium flux are involved in inhibition of insulin release by DPH using in vitro perifusion system of rat isolated islets of Langerhans.
Results are as follows:
1) DPH (1μg-25μg/rni) inhibits insulin release induced by glucose (16.7mM) dose-dependently.
2) The inhibition by DPH is reversible and the rebound occurs. after withdrawal of DPH.
3) The inhibition of insulin release by DPH is reversed by suppression of Na efflux with ouabain and acceleration of Na influx with gramicidin. These findings suggest that DPH inhibits insulin release by suppression of Na influx as well as acceleration of Na efflux by activation of Na+-K+ ATP-ase.
4) The inhibition of insulin release by DPH (5μg/ ml) is reversed significantly by increased Ca influx with high calcium (11.8 mEq/L) or A 23187 (10-5M).
The above data suggest that inhibition of Na net uptake by DPH suppresses Ca influx, and decreased cytosolic calcium concentration thereby produced inhibits insulin release.

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471 episodes of myocardial infarction in 452 diabetic patients (male 308, female 144) who were treated in Japan between January, 1973 and June, 1975 was studied regarding their relationship between early death from myocardial infarction and therapeutic methods of diabetes before the attacks. The data of these patients were collected from members of Japan Diabetic Society in 379 hospitals and clinics.
The episode number of myocardial infarction in non-treated diabetics and diabetics treated with diet only, oral hypoglycemic agents and insulin was 105, 109, 197 and 54, respectively. There was no remarkable differences in age distribution and the peak of frequency was found in age 60-69 year among the all groups.
Among patients treated with diet only, the mortality within 4 days after the attack of myocardial infarction was 15% and the mortality for patients on therapy with oral hypoglycemic agents and insulin was 23% and 28%, respectively. There was no remarkable differences in the mortalities in the early stage (within 4 days) after myocardial infarction among oral therapy and insulin therapy group. The early deaths were commoner in the patients with long duration of diabetes (over five years) and in the elderly patients (over 70 years old).
The mortality one month after myocardial infarction was 48%, 30%, and 26% for the patients treated with insulin, oral hypoglycemic agents and diet only, respectively. In insulin therapy group, the mortality at one month was higher than in other two groups.
The main causes of death within 4 days after the attack of myocardial infarction were left ventricular failure and ventricular fibrillation.

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A case of diabetic Charcot joint with amyotrophy was reported. 63-year-old woman with diabetes of 16 years' duration has been treated with insulin, but poorly controlled. For 3 years prior to this admission, she had developed increasing weakness of both legs and wasting of proximal muscles. Four months before admission she noticed the swelling of the left foot, two months later an ulceration developed on the tip of the left fourth toe.
On examination, proximal muscle atrophy were prominent. EMG revealed myogenic changes in proximal muscle. Histology of quadriceps showed scatterd atrophy. Marked swelling with moist ulcer on the tip of the left fourth toe was observed. X-ray film revealed marked destruction of articular surface as evidenced by irregularity, sclerosing and narrowing of metatarsophalangeal joint surface involving the first, second and third toes. In the fourth toe, absorption of metatarsal bone and phalanx, suggesting of osteomyelitis was observed.

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An epidemiological study of diabetic children in Okayama prefecture was performed. Questionnaires were mailed to all hospitals professing internal medicine or/and pediatrics and to all primary, junior high and high schools in the prefecture. Answeres were obtained from 51.6% of the hospitals and from 90.0% of the schools.
Twenty one diabetic children-10 males and 11 females-were registered. An incidence of diabetes was about 7 per 100, 000 pupils.
There was increased incidence in autumn and winter in 14 cases. In other 7 cases, diabetes was found out by urine examination in their schools. Five out of 21 cases were considered to be adult onset type and possessed either a familial history of diabetes and/or a history of obesity. However, eight out of 16 juvenile type diabetics had not these histories and other two cases of juvenile diabetics developed the disease following infectious illness.
The seasonal distribution of the incidence, abscence of diabetic family histories and preceding infections probably suggest that the onset of juvenile type diabetes might be induced by viral infections.
Fifteen cases were insulin dependent, 3 cases were administered oral agents and other 3 cases were well controlled with diet alone.

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The purpose of the present investigation is to be established a more specific method for the determination of a property of insulin binding antibody in human serum and its clinical evaluations. As the principle of this techinque, double antibody precipitation technique using ¹²⁵I-insulin (porcine) was employed. This method is applicable to only 50 μl of serum of each dilution step (IgG=1: 16, IgA=1: 4, IgM=1: 1, Kappa, Lambda=1: 16) for the determination, and also relatively smaller amount of the second antibody is sufficient to proceed the assay system. Furthermore, it revealed that this method is superior to the other methods (i. e. PEG, gelfiltration, electro phoresis method) in terms of specificity, reproducibility and sensitivity.
The property of insulin binding antibody were observed clinically as followings.
(1) Clases of immunoglobulins: IgG-insulin binding antibodies were detected in 61 patients out of 72 diabetics with insulin treatment and in 3 patients out of 155 hyperthyroidism with thiamazole treatment. However, IgA, IgM-insulin binding antibodies were detected in none of the patients.
(2) Types of light chains of immunoglobulins: Forty-three patients out of 45 diabetics with insulin treatment showed to have both K-and L-type of light chains, but only 2 patients showed K-or L-types. One case of hyperthyroidism with insulin autoimmunity had K-type only.
(3) ¹²⁵I-insulin binding antibodies showed various affinities to the native human, procine and bovine insulin, respectively. However, sera of 9 patients out of 61 with ¹²⁵I-insulin binding antibody did not show affinities to various native insulins, but only showed high affinity to the iodinated insulin.
It is suggests that this unique antibody have a high affinity to an altered insulin by means of the iodination.

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Sulfhydryl reagents such as organic mercurials, disulfide compounds and iodoacetate derivatives with a limited ability to penetrate the B cells have been well known to enhance glucose-induced insulin release even at concentrations lower than 1 mM.
In order to investigate the participation of sulfhydryl groups in the plasma membrane in the release of insulin, we studied the effects of D, L-methionine methyl sulfonium chloride (MMSC) on glucose-induced insulin release and also of urea analogs on the B cells treated with MMSC using the perfused rat pancreas. The results are as follows.
1. Effect of MMSC on blood glucose of mice: 0.1 ml and 0.2 ml of 200 mg/ml of MMSC solution, and 0.2 ml of saline solution as a control were injected intravenously from the tails of 24 hr-fasted mice. Blood glucose levels of mice treated with saline solution were 142±5mg/dl, (n=9, M±SEM) for a 24 hr-fed group, and 91±5 mg/d/(n=9) for a 24 hr-fasted group. Those of mice treated with MMSC solution were 183±9mg/d/(n=24) for a 24 hr-fed group and 121±6mg/dl (n=5) for a fasted group (MMSC; 0.2 ml), and 146±13 mg/dl (n=5) for a 24 hr-fed group (MMSC; 0.1 ml).
2. Inhibitory and enhanced effects of MMSC on glucose-induced insulin release from the perfused rat pancreas: Insulin release stimulated with 16.7mM glucose was biphasic, and the mean rate of insulin release during the first 6-min period and the maximal value were 90±12 ttU/ml/min, and 122±20μU/ml, respectively, The amount of insulin release in the second phase was kept at 130-150μU/ml. Dynamics of insulin release elicited with a combined infusion of 16.7 mM glucose and 10 mM MMSC following after a 15-min perfusion with 16.7 mM glucose showed a transient enahncement (maximal value; 226±6μU/ml) and then a gradual decrease (102±12μU/ml at 35 min). In the case of 0.5 mM MMSC, dynamics of insulin release did not chauge as compared to the control. MMSC at 10mM did not induce insulin release in the presence of 2.8 mM glucose (the mean rate of insulin release; 18±3 μU/ml/min). The amount of insulin release with 16.7mM glucose from the pancres preperfused with 2.8 mM glucose and 10mM MMSC for 10 min decreased to 47-50% of the control both in the first and second phases, and the mean rate of insulin release and the maximal value were 36±10 μU/ml/min and 53±16 μU/ml, respectively.
3. Recoverable and protective effects of urea analogs and nicotinamide on MMSC-treated pancreas: Dynamics of insulin release stimulated with 16.7 mM glucose from the pancrefas treated with a combination of 2.8 mM glucose and 10 mM MMSC for 10 min, and then of 2.8mM glucose and 20 mM urea analogs for 15 min were significantly improved both in the early and second phases as compared to that without treatment of urea analogs. The mean rates of insulin release and the maximal values were 87±8μU/ml/min, 107±19μU/ml for urea treatment, anb 101±21μU/ml/min, 140±11μU/mi for ethyleneurea treantment, respectively. The mean rate of insulin release and the maximal value stimulated with 16.7 mM glucose, after 10 min-preperfusion with 10 mM MMSC and 20 mM nicotinaimde in the presence of 2.8 mM glucose, were 55±7 pU/ml/min and 97±2μU/ml, respectively. We concluded that MMSC may act on the B cell to inhibit and enhance insulin release through the mechanism of alkylation to the thiol groups, and urea analogs may improve insulin release from the MMSC-treated pancreas by unmasking the dithiol groups present at glucoreceptor sites.

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As an approach to the study of prediabetes, women who delivered large babies were examined with respects to their characteristics; family history of diabetes, past history of pregnancy and delivery, glucose tolerance, insulin reaction and other aspects.
1) The subjects studied were 284 mothers of large babies weighing more than 4, 000 g at birth reported in Suita City and its adjacent area (Osaka Prefecture), where total population was about 350, 000. The examinations were performed exactly one year after the birth. The subjects were tended to be more obese than the control group of a comparable age distribution. The incidence of family history of diabetes was answered in 19.8% of the subjects, this rate being very close to that of diabetic cases among diabetes families. It was also found that abortions, stillbirths and delivery of large babies in the past were frequent in this subjects.
2) The rate of diminished glucose tolerance in the subjects did not exceed largely that of the control group. However, age of mother, birth weight of the child, birth order of the child and the past history of large babies related to the rate, indicating the diabetogenic effect of these factors.
3) The insulin response to glucose was evaluated by setting an index; a ratio of area of IRI over that of glucose during glucose tolerance test (EIRI/EBS). The mothers of large babies revealed a decreasing trend of the index compared with the control group among those with normal glucose tolerance. Age of mother, birth order of the child, abortions, stillbirths and delivery of large babies in the past also related to the decrease of the index, suggesting relationship between these factors and deminished insulin reaction.
As a result, mothers who delivered large babies were found to be highly probable to be prediabetic as judged from characteristic features presented here. A long term follow-up study on this subjects should be made in the future.

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Plasma glucagon levels were studied in 11 patients with acromegaly and in 14 patients with isolated growth hormone deficiency. As controls, 20 normal sudjects and 7 normal children were examined. Acromegalic patients were divided into two groups ; one with diabetes mellitus, and the other without diabetes mellitus.
After overnight fasting and absolute bed rest for at least 30 min, all subjects received an intravenous infusion of 30 g or 0.6g/kg of L-arginine over a period of 45 min, All acromegalic patients and 12 normal subjects were also given orally 50 g of glucose. Plasma glucagon was determined by the talcum radioimmunoassay, using antiserum 30 K which is specific to pancreatic glucagon.
Fasting plasma glucagon levels in patients with acromegaly and isolated growth hormone deficiency were 18138pg/ml, 175+21pg/m/, respectively, which were significantly higher than those in the control subjects. In acromegalic patients, plasma glucagon response to arginine was significantly higher than in normal subjects. Plasma glucagon increase induced by arginine was not significantly higher in acromegalic patients with diabetes mellitus than in those without diabetes mellitus. Arginine-induced glucagon secretion in patients with isolated growth hormone deficiency was significantly higher than in normal children. This indicates that both excessive secretion of growth hormone and growth hormone deficiency cause glucagon hypersecretion.
Following oral glucose loading, plasma glucagon decreased successively in normal subjects, whereas it did not decrease and rather tended to be paradoxically elevated in patients with acromegaly. This phenomenon was more remarkable in the patients with diabetes mellitus.
These results suggest that growth hormone is probably required for optimum function of the islets and that excessive growth hormone will cause excessive glucagon secretion which may contribute to the diabetic diathesis in acromegaly. Since hyperglucagonamia was observed in growth hormone deficiency, some other factors influenced by growth hcrmone may affect glucagon secretion.
The significance and pathogenesis of hyperglucagonemia in these conditions are still unclear at present and must await further clarification.

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We have found that in normal subjects, immunoreactive insulin (IRI) response during either glucose or tolbtamide stimulus, are affected by posture, and the IRI response are greater in the sitting than in the right lateral position. We, therefore, have studied the effects of posture on insulin responses in normal subjects.
1) Gastric emptying rate of glucose, rise of blood sugar level and IRI response 30 minutes after oral glucose load were greater in the right lateral position than in the sitting position.
2) During 25 g-intravenous glucose tolerance test (iv GTT), the pretreatments with atropine (0.5 mg) or propranolol (5 mg) 10 minutes before the tests, minimalized the differences in positional changes of IRI responses. On the other hand, pretreatment with pilocarpine (7mg) or trimatoquinal (0.1 mg) enhanced those differences.
3) Pretreatment with atropine (0.5 mg) or pilocarpine (7 mg) had no significant effect on IRI responses during 25 g-iv GTT in the sitting position.
4) Vasopressin (10 units) strongly supressed IRI response during 25g-iv GTT both in the sitting and in the right lateral position, and worsend the glucose tolerance especially in the sitting position.
5) IRI disapperance curve was not affected by positional changes after intravenous injection of regular insulin (4 units).
We reported previously, that during oral GTT, the peak time of blood sugar and IRI response were more retarded in the left lateral position, and that during tolbutamide test IRI response was greater in both right and left lateral positions than in the sitting position.
Our findings, suggest the view that the gastric emptying rate, parasympathetic nerve and β₂ adrenergic receptcr are the main factors responsible for the positional change in IRI response during oral GTT. On the other hand, changes in pancreatic blood flow and insulin degreading rate have little effect on the change in IRI response.

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A new syndrome with hypoglycemia and enpogenous insulin antibody has been reported by Hirata. as one of autoimmune diseases. In Japan 15 cases of the syndrome have been found. In the present stupy we investigated insulin antibody in a case of this syndrome who showed hypoglycemic attacks several times in March, 1975. The patient has not ever bed insulin injection. Since he was admitted on 26th, March, 1975, the hypoglycemic attacks with sweating, palpitation and headache, have spontaneously improved. In his serum the binding protein with 125I-pork insulin was proved by various kinds of methods, i. e., ethanol precipitation, gelfiltration, Separax electrophoresis, disc gel electrophoresis and two anitbody method of Andersen. The insulin binding protein was insulin antibody which was clearly shown with the immunofluorescent technique.
Serum insulin measured by double antibody method, was 1400 pU/m/ but this value was sham due to the presence of insulin antibody in serum that was known to interfere with a radioimmunoassay of insulin. Insulin binding capacity of the antibody was 48.1μU/m/. insulin by the determination of Andersen's method. This capacity spontaneously decreased to 18.7μU/m/. insulin five months after initial hypoglycemic attack occured. This value became under twoμU/m/ insulin one year after.
It is suggested from this case that antigenicity of endogenous insulin inhuman can appear transiently, although mechanism that produces insulin antibody against endogenous insulin is not clarified. As glucose tolerance test was diabetic in this patient further study is needed to explain the pathogenesis of diabetes mellitus related with appearance of insulin antibody.

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There have been several reports concerning the treatment of diabetic coma and ketoacidosis with continuous small-dose infusion of insulin. But none of them were supported by experimental observations. We have already confirmed the efficacy of small-dose insulin infusion therapy in depancreatized ketotic dogs. Based on these observations, 4 diabetic ketosis, 1 non-ketotic hyperosmolar coma and 1 diabetic coma were treated with continuous small-dose insulin infusion. In each case, 0.5%-1% albumin or 0.7% gelatin was added into insulin solutions to prevent the adsorption of insulin on the glassware during the treatment.
Results of the treatments were very satisfactory and all the patients recovered safely within 10hours. Means (±S. E. M.) of pretreatment blood sugar, rate of insulin infusion and rate of decrease in blood sugar level during treatment were 762±127 mg/dl, 8.8±3.3 U/hr and 115±21mg/dl·Ehr, respectively. Plasma 3-hydroxybutyrate levels decreased rapidly and returned to normal levels within 3 hours. Hypoglycemia was not observed in the course of the treatment. As far as the serum potassium levels were concerned, severe hypokalemia was not observed. But the serum potassium decreased significantly as compared to the pretreatment levels in each case. From these results, the necessity of potassium supplementation is suggested in the cases showing low pretreatment levels of potassium.
Thus, it is confirmed that “small-dose insulin infusion therapy” for the diabetic coma and ketoacidosis is as effective as the conventional large-dose insulin therapy.

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Rheological changes in the blood flow and in the blood cell themselves may contribute to the appearance and evolution of diabetic retinopathy. In relation to the sludging phenomenon of erythrocyte in diabetes mellitus, surface electric charge (SEC) and membrane N-acetylneuraminic acid (NANA) were measured. The SEC and NANA levels of erythrocyte were determined by the colloid titration and by Warren's thiobarbituric acid method, respectively.
The SEC levels of erythrocyte were significantly lower in diabetics without retinopathy (group A)(5.94±0.39×1017/ml packed cell, N=25)(P<0.01) than in controls (6.38±0.45, N=15) and in diabetics with retinopathy (group B)(5.70±0.27, N=17)(P<0.05) than in group A.
The membrane NANA levels of erythrocyte were also significantly lower in group A (0.32±0.10 ttmol/m/ packed cell, N=25)(P<0.05) than in controls (0.39±0.09, N=15) and in group B (0.28±0.05, N=17)(P<0.05) than in group A.
It may be conceivable that the above results contribute to the solution of rheological changes of blood in diabetes mellitus.

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