The antigenicity of Monocomponent (MC)-Lente insulin (Monotard) was studied for 8 to 12 months in 19 diabetics and compared with that of mixed-species commercial Lente insulin preparation (C-Lente insulin) obtained from the market before July 1975.The immunogenicity of the MC-Lente insulin was estimated by separate determination of the insulin binding antibodies (porcine or bovine) and proinsulin-specific antibodies (porcine or bovine) by the polyethylene glycol method.These antibody titers were expressed as insulin antibody or proinsulin-specific antibody indices according to the method of Sebriakova.Also, the total extractable serum insulin was assayed by the polyethylene glycol method of Nakagawa with minor modifications.
Eleven diabetics in the first group had been treated with C-Lente insulin for periods of at least 8 months.Their insulin treatment was then converted to MC-Lente insulin after several determinations of their insulin antibody and proinsulin-specific antibody indices for more than 6 months duration.
The diabetics converted to MC-Lente insulin treatment from C-Lente insulin showed striking reductions in both porcine and bovine proinsulin-specific antibody levels in all cases examined. Low levels of these proinsulin-specific antibody levels after 6 months treatment with MC-Lente insulin remained unchanged throughout MC-Lente insulin treatment of 8 to 12 months.
Significant reductions in porcine and bovine insulin antibody levels were observed after the initiation of MC-Lente insulin treatment in this group.In 3 patients, however, both the porcine and bovine insulin antibody levels increased gradually after 8 months treatment with MC-Lente insulin.The total extractable serum insulin levels in this group were markedly reduced after the switch to the MC-Lente insulin treatment.These low but significant levels of total insulin remained unchanged during the later period of observation.
In the diabetics (8 cases) treated with MC-Lente insulin from the beginning of insulin treatment, porcine or bovine proinsulin-specific antibodies were not detectable throughout MCLenteinsulin treatment of 8 to 12 months.Porcine or bovine insulin antibodies were notdetectable during the initial stage of MC-Lente insulin treatment in most cases of this group. Some of the diabetics (4 out of 8 cases) developed weak porcine insulin antibodies during a later period of observation.The total extractable serum insulin levels in this group remained below the level of detection (100 μU/ml) throughout the observation period in most cases, and only 2 of the 8 cases of MC-Lente insulin treated diabetics showed significant levels of total extractable serum insulin exceeding 100 μU/ml after 6 months of treatment.
Insulin allergy and lipoatrophy were not observed in any case during MC-Lente insulin treatment.
These results suggest that porcine Monocomponent-Lente insulin was less immunogenic than the conventional mixed-species insulin preparation purified by recrystallization, and proinsulinspecific antibody levels were markedly reduced after switching to Monocomponent-Lente insulin treatment.Also, from the beginning, Monocomponent-Lente insulin treatment hardly developed any porcine proinsulin-specific antibody.

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The regulation mechanisms of pyruvate dehydrogenase complexes in the gastrocnemius muscles of rats were studied under the following diabetic conditions: acute diabetes (48 hr after the injection of streptozotocin), chronic diabetes (8 days after streptozotocin injection), insulin treatment and its deprivation. The results may be summarized as follows.
1) The level of total activity of the enzyme complex was decreased only in the case of chronic diabetes.
2) The level of the active form of the enzyme complex was decreased in both acute and chronic diabetes and recovered by insulin treatment.
3) On electrical stimulation of the muscles, the levels of the active form of the enzyme complex were increased in the control, acute and chronic diabetic groups.
4) The contents of pyruvate and the ratios of CoA/acetyl-CoA and ATP/ADP remained unchanged under the diabetic conditions.
These above results suggest that the decrease in levels of the active form of the enzyme complex in under both diabetic conditions cannot be fully explained by the content of pyruvate, or CoA/acetyl-CoA and ATP/ADP ratios.

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Previous studies have demonstrated a higher prevalence of thyroid antibodies in diabetics than in age and sex matched controls. Coexistence of Hashimoto thyroiditis and diabetes mellitus has also been noted by several investigators. However, the prevalence of Hashimoto thyroiditis in juvenile-onset and adult-onset diabetics has not yet been reported in detail.
In the present study, 36 patients with juvenile-onset diabetes mellitus and 255 patients with adult-onset diabetes were examined by careful palpation to determine whether or not they had goiters. The occurrence of thyroid antibodies was also investigated in sera from all patients, using the thyroglobulin-coated and microsome-coated tanned red cell hemagglutination tests as prepared by Fuji Zoki Co., Tokyo. Needle biopsy of the thyroid was performed in some cases of goiter, in order to confirm the diagnosis of Hashimoto thyroiditis.
Of the 36 patients with juvenile-onset diabetics, 4 (11.1%) had diffuse goiter, whereas theadult-onset diabetics with goiter amounted to 35 cases (13.7%). The juvenile-onset diabetics showed an increased frequency of goiter, compared to that for young subjects under 20 yr old previously reported. On the other hand, the prevalence of goiter in adult-onset diabetics was similar to that (11.3%) in 150 patients with no evidence of metabolic or endocrine disease who were studied as “hospital controls”.
The adult-onset diabetics with goiter were predominantly women and their ages were distributed mainly from the 4th to 5th decade. The sex distribution of these patients was thus similar to that for patients with Hashimoto thyroiditis. Moreover, 12 (34.3%) of 35 cases with goiter were found to be positive for antimicrosomal antibody and diagnosis of Hashimoto thyroiditis was established in 13 cases (37.1%). In contrast, the prevalence of thyroid antibodies in adult-onset diabetics with no evidence of goiter was not significantly different from that reported previously for normal subjects. It appears therefore that the occurrence of thyroid antibodies in adult-onset diabetics is strongly indicative of an association of coexistent Hashimoto thyroiditis.
Four juvenile-onset diabetics with goiter were diagnosed as having typical Hashimoto thyroiditis on the basis of histological findings for specimens obtained by thyroid needle biopsy. The prevalence of Hashimoto thyroiditis was approximately 11.1% in patients with juvenile-onset diabetes mellitus. Thus, a high prevalence of Hashimoto thyroiditis was evident in juvenile-onset diabetics compared to that reported previously in young subjects under 20 yr old. The juvenile diabetics with Hashimoto thyroiditis were characterized by having mainly insulin-dependent diabetes and by showing extremely high titers of antimicrosomal antibodies. Moreover, the family histories of these patients revealed that at least one close relative had both diabetes mellitus and goiter.

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The correlation between diabetic retinopathy and cerebral thrombosis was studied in aged patients with diabetic glucose intolerance (N= 127). The following results were obtained:
1) The prevalence of cerebral thrombosis in diabetics with diabetic retinopathy (30/52: 57.7%) was significantly (P<0.01) higher than that in diabetics without diabetic retinopathy (24/75: 32%).
2) Categorical regression analysis revealed that among numerous factors such as age, sex, grade of osteoporosis, hematocrit values, serum triglyceride levels, serum cholesterol levels, presence or absence of hypertension, serum uric acid levels, plasma fibrinogen levels, presence or absence of diabetic retinopathy, insulinogenic index, fasting blood sugar levels, peak blood sugar levels after glucose tolerance, pattern of the glucose tolerance curve, and duration of diabetes, two factors, i.e. the presence of diabetic retinopathy and relatively high levels of serum triglyceride, were significantly (P<0.05) related with cerebral thrombosis, but the others were not.
3) No significant correlation between the severity of diabetic retinopathy and the prevalence of cerebral thrombosis could be detected.
On the basis of these results, it is suggested that an intimate relationship exists between diabetic retinopathy (microangiopathy) and cerebral thrombosis (macroangiopathy), but that the risk factors of these two angiopathies are not entirely the same.

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Clinical application of our improved GLC method for estimating tolbutamide and chlorpropamide levels in the plasma is described.
The procedure was used to examine the plasma of 83 diabetics under treatment with tolbutamide (n=53) or chlorpropamide (n=30). The following results were obtained 1) The correlation coefficients between the amount of drugs administered orally and the basal plasma levels of the drugs were 0.64 and 0.61 for tolbutamide and chlorpropamide, respectively. 2) There was no significant difference in basal sulfonylurea levels between those administered the drugs for more than 10yr and those for less than 5yr. 3) The basal plasma chlorpropamide levels in diabetics using 0.5 g of chlorpropamide were significantly correlated with the degree of obesity (r=0.81, p<0.05). 4) Good responders, i.e. those whose FBS levels decreased withincrease in plasma tolbutamide levels, amounted to 7 in 8 diabetics (88%), 5 in 12 diabetics, (42%), and 1 in 7 diabetics (14%), whose daily doses were 1.0 g, 0.5 g. and 0.25 g of tolbutamide, respectively.
The above findings are discussed according to a proposed pharmacodynamic model involving a two-compartment system.

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KK mice were treated daily with glibenclamide from 3 to 8 months of age. The dosage of this drug was 25-75 μg/day and determined to give negative urine sugar. As control animals, age matched KK mice without this drug and nondiabetic C 57 BL mice were used.
These animals were sacrificed at 8 months of age. The renal tissue was studied morphologically and morphometrically, and the results for the drugtreated KK mice and control animals were compared.
The mice treated with glibenclamide showed a significant decrease in glomerular size (p<0.01) and glomerular basement membrane thickness (p<0.05) compared to the KK mice without the drug, together with a tendency to decrement of the mesangial area. Moreover, there was a linear correlation between fasting blood sugar and glomerular size in the KK mice. However, these decreased morphometric values for KK mice were still larger than those in C 57 BL mice. Accordingly, the origin of renal glomerular lesions in KK mice may be suggestive not only of the diabetic states but also of other unknown factors including genetic ones.
To determine the metabolic state in the renal tissue which probably leads to characteristic glomerular lesions in KK mice, ³H-proline or ³H-lysine was administered intraperitoneally to KK mice without glibenclamide and C 57 BL mice which were 8 months old.
The kidneys were removed from these animals after 24hr and 16 days. The ratio of the radioactivity in the renal cortex, i.e. the 16 day value (cpm)/24hr value (cpm), was greater in the KK mice than the C 57 BL mice.
Such a tendency was more clearly supported by results for grain counts in the glomerular tufts using the radioautographic method.
It appears therefore that after the incorporation of proline or lysine in the renal cortex or glomerular tufts, these amino acids are retained longer in KK mice than in C 57 13L mice. The results thus reflect the development of the characteristic glomerulopathy in KK mice.

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A 46-yr-old Japanese male consulted our clinic on 12 January, 1976, due to anorexia weight loss, upper abdominal pain, and pain in the lower extremities. Diabetes mellitus had been diagnosed in October, 1975. At that time, there was no sign of anemia. (hemoglobin 15.0 g/dl, RBC 404×10⁴/mm³). One gram per day of tolbutamide had been administered from 6 November, 1975, until 12 January, 1976. The patient was pale and had mild jaundice. His liver was palpable 3 cm below the right costal margin.
The laboratory findings on his first visit to our clinic were as follows; hemoglobin 6.7 g/dl, RBC 204×10⁴/mm³, reticulocytes 155‰, WBC 9800/mm³ by normal analysis, and platelets 22×10⁴/mm³. Urine urobilinogen was strongly positive, and the concentration of serum bilirubin was 2.91 mg/dl with 1.64 mg/dl of indirect bilirubin.
Immediately after diagnosis of hemolytic anemia at the first visit, the administration of tolbutamide was terminated and daily injection of 12 units of lente insulin was initiated. The patient was admitted to our center on 28 January, 1976. The direct and indirect Coombs tests were negative, and a drug-induced Coombs test. (using Muirhead's method) was also negative. All quantitative determinations of enzyme activities in the erythrocytes including G 6 PD revealed normal results. The pattern of hemoglobin electrophoresis was not abnormal except for a slight increase of HbF.
We were unable to identify the cause of the patient's anemia. However, the anemia improved gradually after termination of the administration of tolbutamide. It thus appeared that some correlation may have existed between the anemia and tolbutamide therapy in this case.

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