There appeared several reports concerning the plasma levels of PHLA in obese subjects and diabetics. However the results are still conflicting. Therefore the present study was undertaken to measure the PHLA levels after intravenous administration of heparin (Novo Co., Denmark; 0.1mg/kg) on the untreated obese subjects and diabetic patients, and the relationship between PHLA and glucose tolerance, relative body weight, serum lipid levels or serum IRI was investigated.
Our assay technique was as follows. Substrate mixture contained dialyzed albumin (Cohn Fraction V) in 2%, 10mg of Intrafat and 10ml of fresh normal human plasma. The substrate mixture was pre-incubated for one hour at 37°C for activation before incubation with enzyme sources. The incubation mixtures contained 0.2ml of fresh serum, 1ml of activated substrate, and they were incubated for ten minutes at 37°C under gentle shaking. The amount of free fatty acids released during incubation was determined according to the method of Novak and was expressed as pEq FFA /ml/min.
The subjects were divided into obese and non-obese groups and then their PHLA levels were evaluated according to the grade of glucose intolerance. The PHLA level in the obese group with slight glucose intolerance was higher than that in the obese group whose glucose tolerance was normal or whose fasting blood sugar was over 160mg/dl.
On the other hand non-obese subjects revealed no correlation of their PHLA levels to the degree of glucose intolerance.
The PHLA values of obese subjects ten minutes after heparin injection correlated significantly with EIRI (sum of the plasma IRI values during OGTT), whereas they have no correlation with either the degree of obesity or plasma lipid levels.
Along with caloric restriction in five obese subjects, the PHLA levels decreased. This decrease in PHLA was paralleled with the reductions of plasma TG and pre-β lipoprotein.
In the obese subjects, both hepatic and extra hepatic PHLA levels were measured selectively according to the protamin technique described by Krauss et al. It was shown that about 70-80% of the total PHLA in the obese subjects might be of hepatic origin.
These findings suggest that PHLA level seems to correlate with serum insulin concentration and that the increase in hepatic PHLA observed in the obese subjects might be related to the elevated plasma insulin.

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Diurnal levels of blood glucose and serum immunoreactive insulin (IRI) were observed during treatment with dietary management alone (D), with glibenclamide (G), and with lente-insulin (I) in nine newly-diagnosed maturity-onset diabetic subjects. Their fasting blood glucose values before treatment were 199.9+21.1 mg %(Mean ±S. E.).
The study was designed as follows: Patients were initially treated with diet alone, and were then given 5mg of glibenclamide before breakfast for three days. After an interval of four days, of diet treatment only, a subcutaneous injection of 16 U MC lente-insulin was administered for a period of three days. The diurnal cycles were determined on the third day of each treatment.
The integrated blood glucose areas below the curves of the diurnal blood glucose levels were 4, 245+669mg. h%in “D”, 3, 317+384mg. h% in “G”, and 3, 177±552mg. h% in “I”. The blood glucose levels were almost equally lowered in “G” and “I”. The integrated serum IRI areas below the curves of the diurnal circulating serum IRI levels were 187±24μu.h/ml in “D”, 296± 65μu.h/ml in “G”, and 267+ 43μu.h/m/ in “I”. The serum IRI levels in “G” and “I” were significantly higher than in “D”(P<0.05). The circulating IRI levels in “G” were comparable to, or occasionally even higher than those in “I”, although the difference between IRI areas in “G” and “I” was not stati stically significant.
Serum IRI in “G” consists essentially of endogenous insulin flowing through the pancreaticoduodenal vein. When the comparable diurnal IRI levels in the systemic circulation are maintained in “G” and in “I”, the insulin concentration in the portal vein is reasonably assumed to be higher in “G” than in “I”. Since the liver is one of the main target organs of insulin, these findings strongly suggest that the hypoglycemic effect of glibenclamide, at least in the short-term observation, is largely mediated by the augmentation of endogenous insulin secretory capacity.

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In order to investigate pancreatic islet cell function under long-term treatment with oral hypoglycemic drugs, glucose and insulin secretory response during oral glucose tolerance test and intavenous tolbutamide test were determined in 32 maturity-onset diabetics who were not controlled by diet alone and sequential effects of diet and sulfonylurea on islet function were assessed over 7 years of periods.
In the poor responders to sulfonylureas or the poorly controlled diabetics, the insulin secretory response during glucose tolerance test was significantly lower than in good responders or the good controlled diabetics. The poor responders showed clearly impaired insulin response to tolbutamide as well as to glucose.
During long-term treatment, glucose tolerance was markedly improved in 6 patients. This amelioration was associated with a significantly increase in mean insulin release during 2 months of treatment. Although mean insulin output slightly declined after 7 years of treatment, the improvement in glucose tolerance was maintained in most cases and slightly decreased in some cases. Among the patients receiving oral therapy for long term, the insulin secretory response was higher in the good controlled patients than in the poor controlled patients.
From these results, it should be stressed that oral hypoglycemic agents should be employed with more caution than in the past. It is unlikely that the stimulatory effect of the sulfonylureas bring about the exhaustion of the pancreatic 13-cell. Analysis of the relationship between insulin and glucose level suggests that the drug may act in part by increasing the sensitivity of the β-cell to glucose stimulus.

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In 1973, ' 74, blindness in diabetic patients who has been treated in 18 hospitals existed Chugoku area of Japan were investigated. The visual acuity of these patients in this series was no greater than 8/200, and the blindness was not directly due to diabetic retinopathy were excluded from this study.
Results were as follows;
1. The percentage of blindness due to diabetes of all blindness who were visited in these hospitals in 1973, '74 was about 13.3%.
2. The number of females exceeded that of males. 36 were females and 14 males.
3. The incidence of blindness in juvenile diabetes exceeded that of adult diabetes.
4. In majority of these patients, control of diabetes before the development of severe retinopathy was poor, which should seem to accelerate the progression of retinopathy. For the greater part of this period, each 17 patients of these were treated by tablet and insulin, 10 had received either no therapy or treatment to control the diabetes and other 6 were unknown.
5. Confined the adult diabetes, the mean interval from the onset of diabetes to the blindness for tablet treated patients was 7.44±4.15 ys, which significantly less than 11.75±4.15ys of that insulin treated group.
6. The mean age at death for 9 patients was 52 ys, of whom 5 were dead in renal and 2 were cardio-vascular disease.

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Recently, the relationship between Sorbitol Pathway and the pathogenesis of diabetic complications has been noticed. There have been scarcely any reports about urinary sugar alcohols in diabetic patients, since extensive levels of glucose in urine disturb the analysis of urinary sugar alcohols. We have devised the method which can remove the glucose in diabetic urine easily, completely and economically. By this method, 100mg/ml urinary glucose, which is considered to be maximum concentration in diabetic urine, was removed completely from diabetic urine.
The sample of urine which was desalted and evaporated to dryness was dissolved in decarbonated water, then the solution was made pass through the column of Dowex 1X8 (OH-) slowly taking over twenty minutes. The samples of urine treated with this method in normal subjects and diabetic patients were analyzed by gas chromatography and following result was gained.
Each urinary sugar alcohol in normal and diabetic urine was measured. Sorbitol in diabetic urine was well correlated to fasting blood sugar and its correlation function was y=0.094X-0.2079. The urine of diabetic patients having fasting blood sugar more than 140mg/dl contained significantly high urinary sorbitol, but the relationship between urinary sorbitol and diabetic complications was not clearly demonstrated.

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Recently, Coxsackie B4 virus (C-B4) infection has been suggested to be one of the causes of diabetes mellitus. In this communication, C-B4 neutralizing antibody titres in sera were determined in 11 patients including two cases (Case 1, Case 2) with acute insulin-dependent diabetes followed by the almost complete remission.
Case 1 is a male subject aged 26. His condition was initially similar to common cold, being accompanied with thirst and acute body weight loss, then he was admitted to our hospital because of diabetic precoma. On admission, his blood glucose concentration was 588mg/dl, plasma IRI 12μU/m/, plasma IRG 440pg/ml (by Unger 30K), and metabolic acidosis was remarkable. After having had some 40 days' insulin treatment, he underwent complete remission with the restoration to approxymately normal fasting blood glucose concentration and O-GTT pattern which have been lasting up to the present time (for more than one and half years) only on the diet therapy. In case 1, C-B4 neutralizing antibody titre showed as high as 128 on admission, rose to more than 256 after 37 days, however, returned to normal after 6 months. Case 2 is a male subject aged 44. He was also admitted to our hospital due to acute diabetes associated with common cold. On admission, his fasting blood glucose concentration was 286mg/dl and he underwent insulin treatment for 40 days, until he could be controlled only by the diet therapy. No abnormal findings were observed thereafter for more than one and half years except slightly decreased glucose tolerance. C-B4 neutralizing antibody titre in case 2 showed 128 on admission, persisting in the same titre, and was normalized in 3 months.
C-B4 neutralizing antibody titres were determined in other 9 with known diabetes; titres were less than 8 in 7 patients, and 32 and 128 in the remaining two.
Case 1 and 2 developed acute insulin deficiency following common cold in parallel with the increased C-B4 neutralizing antibody titres. The diabetic state was proven to be transient, showing 2 to 3 months duration. Therefore, it seems probable that their diabetic state might be attributed to the reversible injury of Langerhans islets by C-B4 infection.

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Recently, the use of serum instead of whole blood for glucose determinations by AutoAnalyzer has been increasingly frequent. Since the glucose values obtained from the serum are significantly different from whole blood values, it is necessary to establish new criteria in interpreting serum glucose level.
First, serum and whole bloold glucose values were compared by simultaneous determinations using AutoAnalyzer on 113 blood samples. A regression equation between serum (Y) and whole blood (X) glucose values was established: Y=1.166X-7.9 (mg/100 ml). This equation means that the serum values exceed the whole blood values when X is greater than 108 mg/100 ml, and vice versa.
Secondly, a comparison was made between serum glucose values determined by AutoAnalyzer (Y) and whole blood glucose values by the Somogyi-Nelson method (X) on 168 blood samples from 42 glucose tolerance tests. The regression equation estimated was: Y=1.138X 4-0.09 (mg/100 m/), where the serum glucose values are always 14 per cent higer than the Somogyi-Nelson values.
On the basis of these findings, the diagnostic standards recommended by the Japan Diabetic Society based on the whole blood method were adjusted for serum glucose values. In applying these standards, the results based on whole blood and serum glucose values of 42 glucose tolerance tests were compared, leading to a good agreement existing between them.

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The purpose of this paper is to study the glycerol metabolism of hypothalamic obese mice and of hereditary obese mice. The glycerokinase [EC 2.7.1.30] activity in the epididymal adipose tissue of goldthioglucose obese mice and of yellow obese mice (A^y/a), fed for 13 weeks (dynamic stage) or for 24 weeks (static stage), was measured by the method of Newsholme modified by Yugari. The plasma immunoreactive insulin (IRI) levels were also measured in these animals and the relationship between plasma IRI and glycerol metabolism was observed. The following results were obtained.
A glycerokinase activity was detected in the epididymal adipose tissue of normal mice. In the dynamic stage of both groups of obese mice, the glycerokinase activity per mg protein was not elevated, but, glycerokinase activity per 105 adipose cells and per epididymal adipose tissue was elevated several times as high as that in their nonobese or lean littermates. Moreover, glycerokinase activity per 10⁵ adipose cells showed positive correlation with plasma IRI. In the static stage, glycerokinase activity was not elevated in either group of obese mice and no correlation was seen between glycerokinase activity per 105 adipose cells and plasma IRI. In the dynamic stage, a remarkable increase of body weight was observed. It is concluded that hyperinsulinemia seems to activate the glycerol metabolism in adipose tissue of obese mice, resulting in the increased triglyceride synthesis and therefore, the development of obesity. In the static stage, hyperinsulinemia was not marked in obese mice, and glycerol metabolism in epididymal adipose tissue seems to be norm alized.

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We have seen two patients with the infection of cryptococcus neoformans, and the autopsy of these cases revealed necrosis of the islets of Langerhans. In these cases, cryptococcus neoformans appeared to induce insulitis. It was obscure why the cryptococci were preferably present in the intrainsular areas. In order to clarify this question, we designed two animal experiments in which histopathological examination was done in multiple organs, especially in pancreas. Some of cryptococci used in our experiments had rather thick capsules.
In the first experiment, the cocci were injected intravenously in the tail of 3rats. Daunomycin was injected intraperitoneally to reduce the phagocytic activity of the leucocytes of the animals. Fifteen days after the injections, the animals were examined. All 3 rats were found to have the cocci in the blood, but only one of them was confirmed to have the cocci in thepancreas, liver and kidney. In this rat, the cocci were seen in the intrainsular and peninsular areas in 3 of 10 islets, while they were very few in the exocrine tissue. Surrounding these cocci, therewere inflammatory infiltrations, markedly in certain areas, and less markedly in the other areas.Degeneration and necrosis of the islets were observed together with the inflammation.
In the second experiments, 3 rats were laparotomized under ether anesthesia, and cryptococci were injected into the common hepatic artery. In this experiment, granulomas, degeneration and necrososis of the parenchymatous areas adjacent to the islets were observed anda part of the islet itself showed glanulomatous change.
Acording to Prof. Fujita, the microcirculation of the pancreas starts with vas afferens into the network of the thick capillaries of the islets and terminates in the exocrine tissue via the acinar portal vein of the islets of Langerhans. Probably the Cryptococci are caught atthe point of the islets or in vasa efferentia, and they stay there instead of leaving with the blood flow.

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Rabbits were immunized with a-, b-component of bovine insulin, a-component of porcine, Insulin, conventional insulin or monocomponent insulin in order to compare the immunogenicity of these components.
Rabbits immunized with a-component of bovine insulin showed the strongest antibody production in early stage. All rabbits immunized with conventional insulin produced insulin antibodies. On the other hand, 2 out of 5 rabbits immunized with monocomponent insulin produced no antibody. Antibody developed against conventional insulin tended to be stronger than that against monocompo. nent insulin.
Rabbits immunized with b-component of bovine insulin showed glucose intolerane in intravenous glucose tolerance test, but these with a-component of bovine insulin showed no glucose intolerance and exhibited reactive hypoglycemia after glucose load.
It is suggested that a-component of bovine insulin may be most immunogenic, but the antibody developed against it may not affect on glucose tolerance.

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The degree of fat mobilization and ketosis in experimental diabetic animals has been shown to be closely related to the amount of body fat stores. A diabetic patient was studied in this paper, who was markedly ketosis-resistant in spite of the severe insulin-deficiency.
He was a 40-year-old skinny man, suffering from diabetes since 6 years ago. He weighed 29.0 kg and was 162cm tall, showing generalized lipo-and muscular atrophy, cataract and diabetic retinopathy (Scott IIIa). IRI response in OGTT and intensive islet stimulation test indicated that he had almost lost the insulin secretory capacity. During 24 hour starvation urinary ketone bodies were negative and the maximal values of FFA and β-hydroxy butyrate in plasma were 491μEq/1 and 2.5mg/dl, respectively.
After 16 months with insulin treatment, he gained 10.0kg in body weight and 24 hour starvation induced marked ketonuria and ketonemia. The maximal values of FFA, β-hydroxybutyrate and acetoacetate in plasma were 912μEq/1, 26.03mg/dl and 2.81mg/dl, respectively.
There were observed neither characteristics suggesting lipoatrophic diabetes nor Werner's syndrome. The fibroblast culture of biopsy specimens of skin grew well. The secretions of pituitary lipolytic hormones and adrenocortical hormone were within normal limits. Therefore, the marked decrease in the ketogenic capacity before insulin treatment should be attributed to the paucity in the body fat store, which had been presumably brought about by the incidious insulin deprivation.
We propose that such metabolic state should be called “non-ketotic chronic insulin deficiency.”

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A 68-year-old woman had suffered from diabetes mellitus for 13 years which had been poorly controlled. She was required to have her left leg amputated three times for diabetic gangrene.
She was admitted to our hospital this time because of a need of amputation for newly developed gangrene in the right leg. The patient had been on 16-32 units of lente insulin during hospital course, but there were no symptoms and signs due to other hormonal abnormalities or hypof unction of the external pancreatic secretion. 3 weeks following surgery, she expired with cerebrovascular disease.
At autopsy, the pancreas appeared normal in shape and size and weighed 70g. The exocrine acini and ducts in the body and tail of the pancreas were completely replaced by adipose tissue, while the head maintained the normal structure of the tissue. There were collections of lymphoid cells in the tail. No calculi or other obstructive masses were found in the head and the body.
Although the etiology remains unknown, the change in the pancreas seems to be the acquired rather than the congenital. It is possible that the obstruction of the duct which occurred through certain mechanism might lead to the partial atrophy of the exocrine pancreas and finally to the replacement by adipose tissue in that portion.
Of interest is the coexistence of multiple endocrine adenomatosis found in this patient at autopsy i. e., non-B cell adenoma of the pancreatic islet, adenomas of the bilateral adrenal cortex and nodular goiter. However, it seems that there is no definite relationship between the adenomatosis and the replacement by adipose tissue in the exocrine pancreas.

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4g of l-arginine monochrolide was injected intravenously as a 2-min “pulse” in seven normal subjects. The arginine pulse was given 3 times at a 30 min-interval. Blood was withdrawn from antecubital vein into heparinized disposable plastic syringes at intervals of 3-10 min. Plasma insulin was measured by the double antibody immunoprecipitation technique of Hales and Randle. Plasma glucagon was determined by the radioimmunoassay method of Sakurai and Imura, utilizing antiserum 30K. The 2 min arginine pulses repeated three times at a 30-min interval elicited similar plasma glucagon responses: peak plasma glucagon levels following the first, second and third pulses were 252-458pg/ml, 240-493pg/mland 256-456pg/ml, respectively. The peak plasma insulin levels after the first, second and third arginine pulses were 30-65 μU/ml, 28-58 μU/ml and 24-62 μU/ml, respectively which were not significantly different from each other. All peaks of plasma glucagon and insulin occurred within 3-5 min after arginine injection. Blood glucose rose slightly and transiently after each arginine pulse. These results indicate that the arginine pulses evoke similar insulin and glucagon responses which given at a 30 min-interval, therefore, arginine injection method is a useful tool as the glucagon secretion test.

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