The materials for our study consisted of pancreatic specimens collected surgically from tvo patients with a new type of spontaneous hypoglycemia. They possessed insulin-binding auto-antibodies without previous history of insulin injection and suffered from severe hypoglycemic attacks.
Case 1 was a 52-year-old male and had had hypoglycemic attacks for 5 years. In the pancreas excised surgically the number of islets per square centimeter of pancreatic tissue was 440-714 and the proportion of islets in pancreatic tissue was 2.9-5.5%, although the normal ranges of the number of islets and proportion of islets reported by Gepts were 53-649 and 0.51-3.09% respectively. Reduction of granules of B cells, fibrosis of islets and dilatation of capillaries in the islets were significant.
Case 2 was a 61-year-old male, and the pancreatectomy of this patient was carried out after a period of 9 months of spontaneous hypoglycemic attacks. The number of islets per square centimeter of pancreatic tissue was 391-616, and the proporton of illets in pancreatic tissue was 8.3-12.2%. Hyperplasia of islet cells and hypertrophy of islets were significant in Case 2, and islets were filled with well granulated B cells. Scattered stomal infiltration of lymphocytes with germinal center formation was found in the exocrine pancreas.
Although it is difficult to prove that the histological changes of the pancreas of these patients are caused by insulin-autoimmunity, insulin-binding antibodies it; the scrum might stimulate the islets directly or indirectly in these cases. Some of differences of qualitative changes in the islets between Case 1 and Case 2 might be related to the different length of the period of hypoglycemic attacks before pancreatectomy.

View full abstract

We have reported that the serum levels of LCAT activity was high in obese subjects and that it decreased significantly after weight reduction by caloric restriction.
The purpose of the present experiments is to discover the factors which have the most intimate relationship with this reduction of LCAT activity.
The subjects of this study were 8 obese patients.
For the reduction of body weight, all patients were treated with a caloric restriction of 800 cal/day for 2-4 weeks.
Their LCAT activity, body weights and serum lipids were measured before treatment and every one week thereafter.
The results were as follows.
(1) In 6 out of 8 cases, the LCAT activity declined rather sharply within one or two weeks, and reached the lowest levels which were about 20-40% of the values of pretreatment.
(2) Although the body weight in all cases decreased during caloric restriction, there was no significant correlation between the absolute values of loss of body weight and the changes of LCAT activity.
(3) There was, however, significant positive correlations between changes of both TC, FC and LCAT activity.
Although the correlative coefficient between changes of TG and LCAT activity was only slightly lower than that of TC and FC, it was not significant statistically.
When obese subjects were treated by caloric restriction, the LCAT activity decreased similarly with the changes of serum lipids.
It is assumed that the 'oss of body weight itself, that is the reduction of the mass of adiposity, has no direct effect on the reduction of LCAT activity. Rather, the profile of serum lipoprotein metabolism, which is connected with adiposity on the one hand, may be more intimately connected with serum LCAT activity on the other hand.

View full abstract

Cyproheptadine (Cyp.) is a pharmacological compound antagonistic to both histamine and serotonin. In a recent study it was well established that Cyp. acts to increase body weight by stimulating the appetite. Drash, A. et al (1966) has comfirmed that the fasting blood glucose level of young adults receiving Cyp. was lowered irrespective of the blood insulin level (IRI) From these data a primary effects of Cyp. on the cell permeability to glucose in peripheral tissues was suggested.
In this study we examined the effect of Cyp. on carbohydrate metabolism in the rat in vivo and in vitro.
In the present experiment in vivo, the fasting blood glucose, oral glucose tolerance test (blood glucose and IRI), and the intraperitoneal insulin sensitivity test were determined in 10 albino Donryu rats (male, 250-300g) which were administered Cyp. (0.1mg/ 100g/ day) for three days.
Glucose uptake by hemidiaphragm and epididymal adipose tissue excised from rats treated with Cyp. was compared to that of nontreated rats.
Fasting blood glucose were decreased in Cyp. treated rats compared to control rats. However changes of the oral glucose tolerance test and the intraperitoneal insulin sensitivity test after the administration of Cyp. were not significant. On the other hand, the in vitro experiment showed an increasing tendency for glucose uptake and significant augmentation of the insulin effect in Cyp. treate vrats.
From these data it is suggested that Cyp. may promote glucose utilization and enhance the insulin effect in peripheral tissues. Hereafter, clinical application of Cyp. for the treatment of insulin-dependent juvenile diabetics anticipating the above effects were suggested.

View full abstract

The isolated perfused rat liver preparation has been used for study of the effects of prolonged fasting against the action of sulfonylureas and exogenous lactate on hepatic metabolism.
Livers were taken from several male rats of the Wistar strain weighing 200 to 300g. They were made to fast 72 hours prior to use. The volume of the initial recirculating solution used was 250ml of Krebs-Ringer bicarbonate buffer containing 2.5% bovine albumin, 10mM glucose and 25% bovine erythrocytes prepared as reported previously. Tolbutamide was added to the medium after 30 minutes pre-perfusion. However, exogenous lactate was perfused without preperfusion.
1) Glucose production from the added lactate, by the prolonged fasted livers was observed during perfusion. Without adding exogenous lactate, glucose production was found to last only 30 minutes after perfusion. It was found that tolbutamide directly suppressed glucose production from exogenous lactate.
2) Exogenous lactate and tolbutamide markedly suppressed ketogenesis. However, no additive effect for the suppression of ketogenesis was found between exogenous lactate and tolbutamide.
3) The degree of utilization by the livers of added lactate markedly went down with tolbutamide. No differences, however, were found in changes in the Lactate/Pyruvate ratio with or without tolbutamide.

View full abstract

A clinical study was performed in seven patients with insulinoma in order to clarify the correlation between the morphological character of the tumors and the biochemical nature of insulin in the plasma or tumors. “Insulin releasability” of each tumor was estimated on the basis of the numbers of the beta-cells and the capillaries in the tumor tissue. The beta-cells of the tumors were classified into three groups by ultrastructural findings: typical, atypical and mixed types. Four tumors showed typical beta-granules, while the atypical granules were observed in two tumors and the mixed in one tumor. The “insulin releasability” was greater in tumors with the atypical or mixed granules than those with typical granules. An increased response of plasma insulin to tolbutamide or glucagon was observed in tumors with the atypical or mixed granules. However, there were no correlations between the granule types and the insulin response to glucose or arginine. The levels of fasting blood glucose and plasma insulin or the ratio of insulin to blood glucose were correlated with the development of the Golgi apparatus and the granular endoplsmic reticulum. The percentage of “big insulin” was higher in the tumors with atypical granules than those with typical granules. Insulin extracted from the tumors with typical granules rendered a higher immunoreactivity for biological activity compared with that from tumors with atypical granules.
It is concluded that some relationships were observed between the biochemical nature of insulin and the morphological findings of insulinomas. Furthermore, this study also presented various suggestions in the mechanism of insulin secretion.

View full abstract

The effect of purified GLI on insulin secretion from rat pancreas pieces was investigated. A crude preparation of GLI was extracted from the porcine intestine using an acid-alcohol extraction solution. The crude GLI was purified by CM cellulose and then by afinity chromatography. The purity of the last preparation was increased by 170 times, when evaluated on the basis of the relative content of GLI to protein, and contaminations of secretin, pancreozymin and insulin in this preparation were negligible.
The insulin releasing activity of the test agents was evaluated by comparing insulin amounts released from rat pancreas pieces into the medium containing 150 mg % of glucose in the presence or absence of various concentrations of the agents. Net increases in the amount of insulin output brought about by the addition of 10, 50 and 250μEg/ml of GLI were -1.3±34.5, 2.9±54.1 and -17.3±25.8μμg/mg tissue/15 min., respectively; all of which were not significantly increased. On the otherhand, 250μEg/ml of pancreatic glucagon enhanced significantly the glucose-induced insulin release in this experimental system. In conclusion, the GLI preparation purified by affinity chromatography was devoid of the stimulatory activity of insulin release within the concentration employed in this study.

View full abstract

Recently Schlichtkrull and his coworkers produced monocomponent insulin (MC insulin) as a less antigenic insulin preparation. We used MC insulin for the treatment of diabetics requiring insulin therapy and compared it with conventional insulin on antigenicity. Serum antibody levels tended to decrease and total serum insulin levels also decreased significantly in the 13 patients who were converted to MC insulin therapy from conventional insulin. That is to say, the total serum insulin level as a percentage to the level before conversion to MC insulin was 42±8.92%(n=13) at the 6th month and 36.7±10.55% at the 12th month after conversion. After conversion to conventional insulin from MC, serum insulin levels in these patients inclined to increase again.
We found insulin antibody produced by MC insulin in 3 out of 6 patients who had had no insulin antibodies before MC insulin treatment. One of the 3 had been treated with conventional insulin for 1 year several years ago, and the other 2 had never been treated with insulin. In one of these 2 cases treated with MC insulin, the insulin antibody level increased continuously, and in the other it decreased again to a trace level in spite of the continuous administration of MC insulin.
We consider that MC insulin is less antigenic than conventional insulin.

View full abstract

rhis study was performed to discover the mechanism of glucocorticoid-induced hyperinsulinemia. Rats treated with a single dose of dexamethasone (50μg/100g B. W.), as early as 12 hrs after the injection, showed an increase in the fasting plasma insulin to the same extent as rats treated with the steroid for 3 consecutive days. Although the both groups of dexamethasone-treated rats exhibited notable increase in the fasting blood glucose as well, insulin/glucose ratios (1/G) were not identical. When insulin was incubated with fat pads obtained from rats treated with a single injection of dexamethasone, no antagonism was observed on its stimulatory effect on glucose metabolism. Treatment of dexamethasone for 3 days resulted in partial suppression of insulin action in the adipose tissue. In vitro insulin secretion provoked by glucose in the pancreas pieces of the rats was greatly enhanced after treatment with either one or three injections of dexamethasone. The pancreas of these rats also demonstrated a significant increase in insulin secretion by glucagon and aminophylline but not by tolbutamide.
From the above data, it is strongly suggested that glucocorticoid may induce hyperinsulinemia, at least during an early stage, not through the mechanism involved in either hyperglycemia or insulin antagonism of the peripheral tissue but by enhancing the sensitivity of the responsiveness of the beta cells to certain kinds of stimuli.

View full abstract

In 1, 739 diabetic patients who died between 1968-1970 in Japan, the causes of death were studied regarding their relationship to the presumptive age of the onset of diabetes and the age of death. The data of these patients were collected from many doctors in 384 hospitals.
Among the diabetics in which the disease had its onset before 50 years old, the most frequent cause of death was diabetic nephropathy. On the other hand, in those in which the onset was at age 50 or over, cerebrovascular disease was the most frequent cause of death. Particularly, in 66 dead cases who aquired the disease when they were under 25 years of age, the main causes of death were diabetic nephropathy (39 %), diabetic coma (29 %) and infectious diseases of various organs (10 %), and none of them died from cerebrovascular disease, ischemic heart disease, malignant neoplasms and cirrhosis of the liver.
Regarding the relationship between the causes of death and the age of death, in the diabetics who died before 30 years of age the most frequent cause of death was diabetic coma. In the patients who died at from age 30 to 59 and in those died at age 60 or over, the most frequent cause of death was diabetic nephropathy and cerebrovascular disease, respectively. Among 66 dead cases in which the onset was under 25 years old, 64 (97 %) cases died before 40 years old.
A ratio of the duration of diabetes to the expectation of life in the general population of a matched age of the onset of diabetes was calculated for those who died from diabetic coma and diabetic nephropathy. In the cases which died from diabetic coma (159 cases) the mean ratio showed a lower value of 23.3+1.6 %(Mean+SEM) than that of 35.0±1.4%(Mean± SEM) in those which died from diabetic nephropathy (294 cases). In those which died from diabetic nephropathy, the mean ratio in the female (28.9±1.6%, N=136) was significantly lower than that in the male (40.3±2.1%, N=158), (p<0.001)

View full abstract

The effects of ethanol on glucose tolerance were studied in 110 subjects consisting of 96 males and 14 females. All cases were subjected to the oral glucose tolerance test (CGTT) and to the combined administration of ethanol and glucose (AGTT). Results obtained were summarized as follows:
1) Ethanol itself had no effect on the levels of blood sugar and serum insulin (IRI).
2) Both the blood sugar and IRI responses to glucose ingestion were significantly enhanced by the simultaneous administration of ethanol in twenty-five subjects with normal GTT patterns.
3) The rise in blood sugar after glucose ingestion was significantly reduced by the simultaneous administration of ethanol, while the IRI response remained unchanged, in seventeen cases of mild diabetes.
4) Ethanol had no effect on the blood sugar and IRI responses to glucose ingestion in eight cases of moderate diabetes and six cases of sulfonylurea.
5) Among 49 cases with border-line patterns in GTT, 16 cases showed aggravated glucose tolerance after ethanol administration, 18 cases unchanged, and 15 cases improved. There were no differences among those three subgroups in regard to the IRI response, body weight, liver function and in drinking history.
6) Ethanol had no effect on the levels of serum HGH, plasma cortisol, and of lipids in OG TT at least within three hours following ethanol administration.

View full abstract

Radioimmunoassay provided dose response curves which showed linearity by the use of the logistic transformation (Rodbard). This transformation that was applicable to radioimmunoassay should be useful for computer processing of insulin and C-peptide assay. In the present studies, standard curves were analysed by testing the fit of analytic functions to radioimmunoassay of insulin and C-peptides. A program for use in combination with the double antibody technique was made by Dr. Kawamata This approach was evidenced to be useful in order to allow automatic computation of data derived from the double antibody assays of insulin and C-peptides. Automatic corrected calculations of radioimmunoassay data of insulin was obtained to be satisfactory.

View full abstract

From the viewpoint of the pathogenesis of obesity it is of significance to know the relation between the abnormal metabolism observed during obesity and the morphological changes of fat cells.
Human subcutaneous adipose tissue obtained from the abdominal wall during surgery was fixed and isolated by incubation in a 2% osmium tetroxide solution, after which the number and size of the fat cells were ascertained. The total number of fat cells was determined by dividing the body fat by the lipid content of the fat cells. The size of the fat cells was determined by taking the average of the diameters of the fat cells measured under a microscope.
The results obtained were as follows:
On the basis of the size of fat cells, obesity may be classified into two types: obesity with hypertrophied fat cells, and obesity with normal size fat cells. The latter type occurs in children under ten years of age and is characterized by an increase in the number of fat cells. Hypertrophy of the fat cells was observed in most cases of medium or mild obesity.
Comparison of obesity characterized by hypertrophic fat cells and that with normal size fat cells revealed that the former type elicited a high insulin response and carbohydrate intolerance following oral glucose administration, and also showed a higher level of free fatty acids in the blood.
The causal relation between hypertrophy of fat cells and abnormal metabolism associated with high insulin response has not yet been clarified. However, it is thought that the high insulin response observed in the type of obesity with hypertrophic fat cells is a primary phenomenon and that the hypertrophy of the fat cells is due to the influence of insulin.

View full abstract

The relation between the abnormal metabolism of obesity and the morphological changes of fat cells has been previously reported. In the present paper the relation between the morphological changes of human fat cells, particularly their size, and the activation of metabolism of these cells is discussed.
Human adipose tissue was obtained during surgery from both obese and nonobese patients. Activation of metabolism was represented by the amount of glycerol released by the fat cells.
Comparison of the size of fat cells of obese and non-obese subjects revealed that the former was always enlarged. The rate of basal lipolysis of hypertrophied fat cells of obese subjects was higher than that of the normal sized fat cells of non-obese subjects. However, it was found that in the hypertrophic fat cells lipolysis was rather decreased when a high concentration of norepinephrine or dibutyril cyclic AMP was added in order to stimulate lipolysis.
Acceleration of basal lipolysis was closely related to the size of fat cells. When lipolysis was stimulated by the addition of a high concentration of norepinephrine and dibutyril cyclic AMP, there was a tendency for the lipolytic response to become augmented with an increase in size of the fat cells. However, when large fat cells formed the majority of the adipose tissue, the lipolytic response was diminished.
The observation that the lipolytic response was weakened when there was an abundance of hypertrophic fat cells suggests that the activation of lipolysis was inhibited by some factor.

View full abstract

The subjects were 139 diabetics (51: insulin treatment, 71: oral drugs, 17: pretreatment) who had no apparent atherosclerosis, obesity or secondary hyperlipemia except that due to diabetes. The present investigation was focussed on the effects of different ways of treatment of the serum lipid metabolism of diabetics.
The results were as follows:
(1) The mean serum lipid levels (TL, TG, TC) were higher in the oral drug group with poor control than those with good control and insulin groups without regard to their control state. The fasting serum NEFA levels were measured in both oral and insulin groups, and the mean values were slightly lower in the treated than in the pretreated in both groups. However, the difference was not significant.
(2) Compared to other groups, in the oral hypoglycemic group with poor control, the percentage composition of the serum total lipids was increased for palmitic acid and oleic acid and was decreased for linoleic acid. This pattern of the change of the fatty acid composition was also found in triglyceride, esterified cholesterol, NEFA and phospholipids.
(3) The incidence of hypertension and EKG abnormalities was higher in the oral hypoglycemic group with poor control than the other three groups.
Although the clinical status of the oral hypoglycemic group with poor control is somewhat different than the other three groups, the results of our study emphasizes that when the diabetics are treated with oral drugs without significant improvement in carbohydrate metabolism, the serum lipid metabolism also was prone to deviate from normalization.

View full abstract

The clinical course and details of diabetic complications were described for 3 cases of the congenital type of lipoatrophic diabetes among 6 of our series.
Case 1, a 19 year old female, was diabetic on the first admission at the age of 9 and deveoped diabetic retinopathy at 14 an.i albuminuria at 19. Renal biopsy revealed mesangial proliferation and nodular lesion. Peripheral nouropathy was also noted and the maximum nerve conducting velocity was as low as 46 m/sec.=
Case 2, a 13 year old male, a brother of case 1, was found to be diabetic at the age of 13 on the third admission, when albuminuria developed and a diffuse type of diabetic glomerulosclerosis was detected by renal biopsy. No retinopathy or nouropathy was found.
Case 3, a 26 year old female, was already diabetic on the admission at the age 24, having retinopathy of Scott I a and unilateral proliferative change. Peripheral and autonomic neuropathy were also present and the maximum nerve conducting velocity was 40.3m/sec. One year after the admission, albuminuria was noted and a diffuse type of diabetic glomerulosclerosis was detected by renal biopsy.
Since Marcus' report, diabetic complications have been detected in 6 cases of lipoatrophic diabetes, and 5 of the 6 cases were the acquired type.
Our observation revealed that diabetic complications, as seen in primary diabetes mellitus, may develope in lipoatrophic diabetes of the congenital type, supporting the hypothesis that mabnormality of motabolis.n plays an essential role in the development of diabetic complications.

View full abstract

We present a patient with an “insuli: i autoim: nune syndrome”. A twenty one year old woman came to the hospital, suffering from frequently cccured hypoglycemic attacks on June 26, 1973.
Although the patient had never received exogenous insulin, significant insulin-binding antibodies were found in the serum, and a huge amount of immunoreactive insulin (21, 500, uU/ml) was detected from the serum at fasting.
Insulin-binding antibodies showed an affinity to bind human and pork insulin more than bovine insulin.
By radioimmunoelectrophoresis, insulin-binding antibodies were found in IgG. The light chain of insulin-binding globulins of the patient were of the kappa type.
Based on pathogenetic consideration, we started corticosteroid therapy to suppress the production of insulin-binding antibodies. The hypoglycemic attacks of the patient disappeared by the treatment, and after one year of steroid therapy, we could not detect elevated immunoreactive insulin in the serum of the patient.

View full abstract