Morphometric comparison of the motor trigeminal neurons.

Comparative morphological and morphometric studies of the motor trigeminal nucleus (MTN) were carried out on the brains of an alligator, a human being and a rat for the first time. Differences in neurons regarding sizes, areas, numbers, column volumes, neuropil indices and circulatory ratios were found. The results showed that the positions and shapes of the MTN were different, and the shapes of motor neurons were different, too. The average area of motor trigeminal neurons was the largest in the alligator, compared with the area in the human being, and the rat; the number of neurons and the column volume of MTN were the largest in the human being. As for the neuropil indices, the value was in descending order: alligator, human being and rat; whereas for the circulatory ratio of neuronal cell bodies, the order was rat, alligator, and human being. We conclude that the size of motor neurons in the MTN may be related to the power of masticatory muscles, and there may be a connection between jaw sizes and shapes and sizes of the motor neurons in the MTN.

[The diagnosis of a mutation of the insulin receptor by non-radioisotropical RT-PCR-SSCP analysis].

We evaluated a 24 years old diabetes woman with type A insulin resistant (patient "Yakushima") who had some typical symptoms as acanthosis nigricans, hirsutism and virilization. Insulin binding to the patient's erythrocytes was significantly decreased to about 30% of the normal control. In order to determine the mutation of the insulin receptor, we used for reverse transcript-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis without using radiolabeled materials. We also analysed the nucleotide sequence with non-isotropical probe. Our results suggested that the mutation was heterozygous, and the patient had a new missense mutation substituted Asn461 for Thr461 in the alpha-subunit.

Dietary sardine oil increases erythrocyte membrane fluidity in diabetic patients.

The effects of dietary sardine oil rich in eicosapentaenoic acid, C20:5 (EPA), on erythrocyte membrane fluidity and membrane and plasma lipids were investigated in diabetic and control subjects. Before consumption of this oil, the levels of erythrocyte membrane fluidity were lower in the diabetic subjects, as noted in our previous work (Diabetes 1983; 32:585-91). Decreased membrane polyunsaturated fatty acid contents were evident. Daily consumption of 2700 mg of sardine oil for 8 wk increased erythrocyte membrane fluidity, as determined by electron spin resonance using the 12- or 16-stearic acid label. This increase was seen after 4 wk, and the level remained elevated for 8 wk. Membrane EPA of phospholipid acyl-chains significantly increased after 4 wk and was even more apparent after 8 wk. Membrane-free cholesterol to phospholipid molar ratios significantly decreased after 8 wk. Both the diabetic and normal subjects responded to the sardine oil in the same way. After feeding with sardine oil, there no longer were differences in erythrocyte membrane fluidity between the normal and diabetic subjects. We propose that improvement in membrane fluidity may contribute to the amelioration of altered cell membrane functions in diabetic patients.

Metabolic dependence of the fluidity of intact erythrocyte membrane.

The motion of spin labeled phosphatidylcholine embedded in the lipid bilayer of erythrocyte membrane increased in association with metabolic ATP-depletion. The fluidity change was reversed by subsequent ATP-replenishment. However, levels of cellular 1,2-diacylglycerol, GSH or intracellular Ca2+ contributed little to the fluidity change. Temperature dependence of the fluidity indicated the disappearance of inflection points in ATP-depleted cells, thereby suggesting alterations in protein-lipid interactions. The fluidity change was also reproduced with oxidizing agents which cross-link spectrin. We suggest that the lipid phase state of membrane is maintained by protein-lipid interactions which depend on the metabolic state of the cells, particularly ATP levels.

Higher levels of erythrocyte membrane microviscosity in diabetes.

Significantly higher levels of erythrocyte membrane microviscosity (MV) [n-: 5.22 +/- 0.17 (4.70--5.92), mean +/- SD (range), poise, N = 67, P less than 0.005] measured by fluorescence depolarization using 1,6-diphenyl-1,3,5-hexatriene as a fluorescent probe were found in diabetic patients when compared with normal controls [5.05 +/- 0.15 (4.70--5.29), N = 22]. No significant differences in MV existed between males and females, nor was MV significantly correlated with diabetic age, duration of diabetes, plasma cholesterol, cholesterol/phsopholipid ratios, and plasma lecithin:cholesterol acyltransferase activities. No significant difference in MV was observed between groups with or without diabetic retinopathy. There was, however, significantly higher MV [5.29 +/- 0.19 (5.00--5.92), N = 20, P less than 0.05] in the group with fasting blood glucose (FBG) greater than or equal to 140 mg/dl than that [5.19 +/- 0.15 (4.70--5.46), N = 47] in the group with FBG less than 140 mg/dl. The changes in erythrocyte membrane MV presented in this study appear to be related to the current metabolic control of diabetic patients and are considered to be one of the factors responsible for the reduced erythrocyte deformability in diabetes.

The lower levels of erythrocyte surface electric charge in diabetes mellitus.

Lower levels of surface electric charge and membrane N-acetylneuraminic acid of erythrocytes were found in patients with diabetes mellitus. These findings appear to be related to microvascular involvement.