A novel point mutation in the human insulin gene giving rise to hyperproinsulinemia (proinsulin Kyoto).

We have identified a 65-yr-old nonobese Japanese man with diabetes mellitus, fasting hyperinsulinemia (150-300 pM), and a reduced fasting C-peptide/insulin molar ratio of 2.5-3.0. Fasting hyperinsulinemia was also found in his son and daughter. Analysis of insulin isolated from the serum of the proband and his son by reverse-phase high performance liquid chromatography revealed a minor peak coeluting with human insulin and a major peak of proinsulin-like materials. The insulin gene of the patient was amplified by the polymerase chain reaction and the products were sequenced. A novel point mutation was identified in which guanine was replaced by thymine. The substitution gives rise to a new HindIII recognition site and results in the amino acid replacement of leucine for arginine at position 65. These results indicate that the amino-acid replacement prevents recognition of the C-peptide-A chain dibasic protease and results in an elevation of proinsulin-like materials in the circulation. Furthermore, in this family the proinsulin-like materials is due to a biosynthetic defect, inherited as an autosomal dominant trait. Rapid detection of this mutation can be accomplished by HindIII restriction enzyme mapping of polymerase chain reaction-generated DNA, which enables us to facilitate the diagnosis and screening.

Effects of Aerobic and Resistance Training Combined with Fortified Milk on Muscle Mass, Muscle Strength, and Physical Performance in Older Adults: A Randomized Controlled Trial.

OBJECTIVES: Fortified milk and resistance training (RT) increase muscle mass, muscle strength, and physical performance in older adults, but it remains unclear whether RT combined with aerobic training (AT) would have stronger effects on these outcomes. The purpose of this study was to examine the effects of aerobic and resistance training (ART) combined with fortified milk consumption on muscle mass, muscle strength, and physical performance in older adults. DESIGN: Open-labeled randomized controlled trial. SETTING: University of Tsukuba. PARTICIPANTS: Fifty-six older adults aged 65-79. INTERVENTION: Participants were randomly allocated into resistance training (RT + fortified milk, n = 28) and aerobic and resistance training (ART + fortified milk, n = 28) groups. All participants attended supervised exercise programs twice a week at University of Tsukuba and ingested fortified milk every day for 12 weeks. Skeletal muscle index ([SMI]: appendicular lean mass/height2) was assessed using dual-energy X-ray absorptiometry as a muscle mass measure. One-repetition maximum strength was measured using four kinds of resistance training machines (chest press, leg extension, leg curl, and leg press) as muscle strength measures. Sit-to-stand and arm curl tests were also assessed as physical performance measures. MEASUREMENTS: The primary measurements were muscle mass and strength. The secondary outcomes were physical performance, blood samples, habitual diet, habitual physical activity, and medication use. RESULTS: Although the muscle strength and physical performance measures significantly improved in both groups, SMI significantly improved in only the RT group. There was no significant difference in the change in SMI and muscle strength measures between the two groups. However, the change in sit-to-stand and arm curl measures in the ART group were significantly higher than those in the RT group. CONCLUSIONS: These results suggest that AT before RT combined with fortified milk consumption has similar effects on skeletal muscle mass and strength compared with RT alone, but it may be a more useful strategy to improve physical performance in older adults. Although the mechanism of our intervention is uncertain, our program would be an effective prevention for sarcopenia in older adults.

Hypolipidemic effects in dogs of ML-236B, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase.

ML-236B, a competitive inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase, significantly reduced both serum cholesterol and phospholipid levels in dogs, when used at a dosage higher than 10 mg/kg per day. Triglyceride levels were not consistently changed, but beta- and pre-beta-lipoproteins were preferentially reduced. Serum cholesterol levels were reduced by 44--45% at the higher dosage of 100--400 mg/kg per day (for 5 weeks) but ML-236B caused no significant changes in the cholesterol content of the liver and aorta and in the activities of serum GOT, GPT, CPK and lecithin : cholesterol acyltransferase. Fecal excretion of neutral sterols was unaffected but that of bile acids was markedly elevated by the drug. Under these conditions, hepatic cholesterol 7alpha-hydroxylase, the rate-limiting enzyme in bile acid biosynthesis, showed no detectable changes.

Studies on anticoccidial agents. 11. Synthesis and anticoccidial activity of nitropyridinecarboxamides and derivatives.

Of the nine nitropyridinecarboxamides, which are isomers of 5-nitronicotinamide, a potent anticoccidial agent, 2-nitropyridine-3, -4, -5-, or 6-carboxamides and 3-nitropyridine-4- or 6-carboxamides were prepared from the corresponding acids via the esters of the acid chlorides. 3-Nitropyridine-2-carboxamide was obtained from 2-methyl-3-nitropyridine by oxidation with SeO2, oximation, dehydration with Ac2O, and hydrolysis with H2SO4. 4-Nitropyridine-2-carboxamide was prepared from 2-cyano-4-nitropyridine by hydrolysis, and the 3-carboxamide analogue was obtained from 4-amino-3-cyanopyridine by oxidation with H2O2 and fuming H2SO4. Of these compounds 2-nitro- and 3-nitro- but not 4-nitropyridinecarboxamides were found to be active against Eimeria tenella. N-Substituted analogues of 2-nitro- and 3-nitropyridine carboxamides were also prepared in a conventional manner and optimal anticoccidial activity was attained with 2-nitroisonicotinamide and its N-alkanoyl, N-aromatic, and N-heterocyclic acyl derivatives.

Studies on anticoccidial agents. 10. Synthesis and anticoccidial activity of 5-nitronicotinamide and its analogues.

5-Nitronicotinamide (1) was prepared from 5-bromonicotinoyl chloride by treatment with ammonia and then oxidation with fuming H2SO4 and 30% H202. 2-Cholor-, 2-alkoxy-2-benzyloxy,2-phenoxy-,2-alkylamino-, and 2-benzylamino-5-nitronicatinamides were also prepared via 2-chloro-3-cyano-5-nitropyridine. 2-Methyl-5-nitronicotinamide (2) was obtained from ethyl 2-methyl-5-nitronicotinate by treatment with ammonia; the 4-methyl analogue 3 was from 3-cyano-2,6-dihydroxy-4 methylpyridine by nitration, chlorination, and dechlorination, and the 6-methyl analogue 4 was prepared by transforming 2-chloro-3-cyano-6-methyl-5-nitropyridine to the corresponding amide, followed by dechlorination. Of these compounds, the 5-nitronicotinamide, the 2-methyl and 4-methyl but not the 6-methyl, analogue showed significant anticoccidial activity against eimeria tenella. N-Substituted analogues of 5-nitronicotinamide and 2-methyl-5-nitronicotinamide were prepared in a conventional manner and optimal anticoccidial activity was attained with their lower N-alkyl analogues, N-alkanoyl and -alkenoyl analogues, and N-aromatic acyl analogues together with these parent compounds.

Studies on anticoccidial agents. 13. Synthesis and anticoccidial activity of nitropyridine-2- and -3-sulfonamides and derivatives.

Eight nitropyridinesulfonamides and pyridinesulfonamide N-oxides as their bioisosteres were prepared and evaluated for anticoccidial activity. Of these compounds, 2-, 4- and 5-nitropyridine-3-sulfonamides and pyridine-2- and -3-sulfonamide N-oxides were found to be active against Eimeria tenella. Thus, the relative positions, ortho or meta, of the substituents in nitropyridine-3-sulfonamides and pyridinesulfonamide N-oxides are important for anticoccidial activity. N-Substituted analogues of 5-nitropyridine-3-sulfonamide were also prepared and optimal anticoccidial activity was attained with the sulfonamide and its lower N-alkyl derivatives. The mode of action of 5-nitropyridine-3-sulfonamide was examined and found to be active in the sporozoite and the first schizogony stages.

Studies on anticoccidial agents. 12. Synthesis and anticoccidial activity of methyl-2(6)-nitro- and -3(5)-nitropyridinecarboxamides.

A number of methyl-2- and 3-nitropyridinecarboxamides have been synthesized. It has been established that the presence of at least one hydrogen atom, adjacent to the NO2 function, is important for anticoccidial activity and introduction of a methyl group to the adjacent position of the CONH2 function sometimes confers enhanced activity. Among the compounds herein, 5- and 6-methyl-2-nitropyridine-4-carboxamides possess optimal anticoccidial activity, being as potent as the parent compound.

Characteristic features of insulin secretion in the streptozotocin-induced NIDDM rat model.

Male Wistar neonatal rats at age 1.5 days (Streptozotocin [STZ] group 1) and 5 days (STZ group 2) received a subcutaneous injection of 90 mg/kg STZ. After 10 weeks, the rats were subjected to an oral glucose tolerance test (OGTT) (2 g/kg) in a conscious state. The pancreas perfusion experiments were conducted 2 weeks after the OGTT. There was no statistical difference in insulin response between the STZ group 1 and the control group. On the contrary, in the STZ group 2, the plasma glucose response to OGTT showed a typical diabetic pattern, and the plasma insulin response was markedly blunted. In the isolated perfused rat pancreas, the infusion of glucose evoked a biphasic insulin secretion, but the peak insulin levels induced by 16.7 mmol/L glucose in the STZ group 1 were significantly lower than in the controls. We further investigated characteristics of insulin secretion in response to different secretagogues in these animal models using isolated islets. The insulin content of the islets of the STZ group 1 were about one half that of the control group. Insulin secretion in the STZ group 1 was impaired in response to glucose stimulation, but remained normal in response to arginine and forskolin. These results suggest that insulin secretion of non-insulin-dependent diabetes mellitus (NIDDM) rat model is selectively impaired in response to glucose stimulation, possibly due to a disorder of signaling mechanism other than adenylate cyclase.

Comparative effect of somatostatin-14 and somatostatin-28 on glucagon-induced glycogenolysis from the perfused rat liver.

The effect of somatostatin (SS)-14 and SS-28 on glycogenolysis was studied, using a rat liver perfusion technique. Livers from nonfasted rats were perfused with 5.5 mmol/L glucose or perfusate without the glucose addition. Glucagon-induced glucose output was lower in the presence of 5.5 mmol/L glucose than in glucose free perfusate at every concentration of glucagon. Under glucose free conditions, SS-14 given at five minutes prior to the glucagon addition reduced the glucagon-induced glucose output dose-dependently. SS-14 given 15 minutes after glucagon addition also inhibited glucagon-induced glucose output significantly. However, various concentrations of SS-28 failed to affect glucose output. On the other hand, in the presence of 5.5 mmol/L glucose, neither SS-14 nor SS-28 affected glucagon-induced glucose output. It is suggested, therefore, that glycogenolysis induced by glucagon from the liver is reduced by SS-14, but not by SS-28, only under glucose free conditions.

Effects of naloxone on basal and vagus nerve-induced secretions of GRP, gastrin, and somatostatin from the isolated perfused rat stomach.

The effects of naloxone, an opiate antagonist, on basal and vagus nerve-induced secretions of GRP, gastrin, and somatostatin were examined using the isolated perfused rat stomach prepared with vagal innervation. Naloxone (10(-6) M) significantly inhibited basal somatostatin secretion in the presence and absence of atropine and of hexamethonium, whereas basal GRP and gastrin secretion was not affected by naloxone. Electrical stimulation (10 Hz, lms duration, 10V) of the distal end of the subdiaphragmatic vagal trunks elicited a significant increase in both GRP and gastrin but a decrease in somatostatin. Naloxone (10(-6) M) failed to affect these responses in the presence or absence of atropine. On the other hand, when hexamethonium was infused, naloxone significantly inhibited both the GRP and gastrin responses to electrical vagal stimulation. Somatostatin secretion was unchanged by vagal stimulation during the infusion of hexamethonium with or without naloxone. These findings suggest that basal somatostatin secretion is under the control of an opiate neuron and that opioid peptides might be involved in vagal regulation of GRP and gastrin secretion.

Effect of vitamin D on gastrin and gastric somatostatin secretion from the isolated perfused rat stomach.

We have studied the role of vitamin D in the regulation of gastrin and gastric somatostatin secretion from the isolated perfused rat stomach. In Ca-deficient vitamin D-deficient rats (Ca(-)D(-) group), the basal and bombesin-stimulated gastrin and gastric somatostatin release (basal IRGa, basal IRS, sigma delta IRGa, and sigma delta IRS) all were significantly lower than in Ca-replete vitamin D-replete rats (Ca(+)D(+) group), and also lower than in Ca-replete vitamin D-deficient rats (Ca(+)D(-) group) except for the basal IRGa. In the Ca(+)D(-) group, the basal IRGa and IRS, and sigma delta IRS were not significantly lower than in the Ca(+)D(+) group. Although there was no significant impairment in basal IRGa, sigma delta IRGa in the Ca(+)D(-) group was significantly lower than in the Ca(+)D(+) control group. Thus, the gastrin and gastric somatostatin secretion from the Ca-deficient vitamin D-deficient rats were impaired. In addition, the impaired gastrin and gastric somatostatin secretions seem to be caused not only by a decrease in serum Ca but also by the reduced effect of the vitamin D on the G and gastric D cells.

Pholipomycin, a new member of phosphoglycolipid antibiotics. III. Biological properties.

Pholipomycin, a new member of the phosphoglycolipid antibiotics, was primarily active against gram-positive bacteria including clinically isolated resistant bacteria. It differed from other members of the antibiotic family as it also demonstrated activity against gram-negative bacteria. Pholipomycin protected mice from infection with Staphylococcus aureus and was nontoxic to mice having an LD50 (i.v.) of 600 mg/kg. Besides possessing anti-microbiol activity, pholipomycin, when administered orally, appeared to promote growth in chickens and pigs.

Evaluation of body composition using dual-energy X-ray absorptiometry, skinfold thickness and bioelectrical impedance analysis in Japanese female college students.

We compared three methods for evaluating body composition: dual-energy X-ray absorptiometry (DXA), skinfold thickness (Skinfolds), and bioelectrical impedance analysis (BIA). Subjects were 155 healthy young college-aged Japanese females whose mean+/-SD (range) age, body height, body weight and body mass index (BMI) were 20.1+/-0.3 (19.6-21.1) y, 158.9+/-4.7 (145.4-172.6) cm, 52.0+/-6.8 (39.4-84.6) kg and 20.6+/-2.3 (16.5-32.5), respectively. Their mean skinfold thickness at the triceps and subscapular were 16.9+/-4.7 (8.0-31.0) and 16.0+/-5.7 (7.0-40.0) mm, respectively. Mean body fat mass percentages evaluated by DXA, Skinfolds and BIA were 29.6+/-5.1, 22.8+/- 5.3 and 25.8+/-4.7%, respectively. Body fat mass was 15.4+/-4.4, 12.1+/-4.5 and 13.6+/-4.5 kg, respectively. Simple correlation coefficients between the three methods for body fat mass percentages provided the following coefficients: r=0.741 for DXA vs. Skinfolds, r=0.792 for DXA vs. BIA and r=0.781 for Skinfolds vs. BIA. Simple correlation coefficients for body fat mass were as follows: r=0.898 for DXA vs. Skinfolds, r=0.927 for DXA vs. BIA and r=0.910 for Skinfolds vs. BIA (all p<0.001). There were significant differences in the values among the three methods with the Skinfolds providing the lowest body fat mass and percentage, and DXA the highest (p<0.001). They all appear to be strongly correlated for evaluating body composition: however, different cut-off values for defining obese and lean need to be defined for each method.

[Awareness of the health and defecation tendencies among college students by location of domicile].

In order to compare the food habits and defecation tendencies by region, a questionnaire survey was conducted of 1722 college women (18-20 years of age) living in Sapporo (146), Sendai (183), Osaka (1214), and Kumamoto (179). (1) When questioned about food habits, less than 58.5% of the respondents in the four groups considered their food intake to be sufficient to maintain their health. In regard to knowledge about their food intake requirements, the positive response was the highest in Sendai (85.8%), followed by Kumamoto (58.2%) and Osaka (50.9%) with the lowest in Sapporo (41.1%) (p < 0.01, among the four groups). (2) The students in Sendai, Osaka, Kumamoto and Sapporo, 96.2%, 93.5%, 92.9% and 92.5%, respectively, considered that constipation could affect their health (p < 0.05, among four groups). The percentage of those with regular bowel movements every day, tended to decrease in the order of Osaka (22.2%), Sapporo (21.2%) and Sendai (20.8%), with the lowest in Kumamoto (12.3%). Bowel movements of less than three times a week were reported by 24.7% in Sapporo, 24.0% in Sendai, 23.2% in Osaka and 22.3% in Kumamoto (p < 0.01, among the four groups). (3) Of those who defecated every day, in terms of time of day, the percentage was high between waking and after breakfast, with the highest percentage in Kumamoto (72.8%) and the lowest in Sapporo (61.2%) (p < 0.05, among the four groups). For subjects that responded that they should have a bowel movement every day, the highest percentage was in Osaka (91.0%) and the lowest was in Sapporo (83.9%) (p < 0.05, among the four groups).