Electron microscopic and pharmacological studies on the rat median eminence

Summary The rat median eminence contains at least three kinds of granules or vesicles: 1. large electron-dense granules (perhaps carriers of neurohypophysial hormones), 2. small electron-dense granules with or without haloes (perhaps carriers of catecholamines) and 3. synaptic vesicle-like structures (perhaps carriers of acetylcholine). The former two electrondense granules exist in separate axons but they coexist with the latter vesicles in the same axons.The pars nervosa shows basically a similar structure to the median eminence. However, the axons containing the small electron-dense granules are very few. In the pars tuberalis, there are at least two types of cells: the cells of one type contain much cytoplasm with large round nuclei and those of the other type contain a small amount of cytoplasm with polymorphic nuclei. The cells of the former include multivesicular bodies and secretory granules, but those of the latter do not. Some of capillaries of the primary plexus are surrounded by the cells of the pars tuberalis on one side and by neurosecretory axon endings on the other side.The median eminence contains high concentration of acetylcholine or an acetylcholine-like substance and shows neurohypophysial hormone activity.Aided by Grant A-3678 from the United States National Institute of Arthritis and Metabolic Diseases. The authors are indebted to Dr. Welsh, Harvard University, for the kind gift of Mytolon.     

Methods for estimating the number of the Cryptomeria red mite,especially with the removal by solutions

We compared the removal by solution, the represented count-area method and the beating, for the purpose of estimating the number of the Cryptomeria red mite. Among them the solution procedure provided the smallest standard error as per cent of the mean. 0.25 per cent unheated solution of sodium hydroxide is used for the summer generation, and also 0.25 per cent boiling one for the winter eggs. The mean proportion removed±standard error for the summer eggs and the winter eggs were 0.8770±0.0316 and 0.7920±0.0281 respectively, while 0.9894±0.0050 for the mites.     

Heterogeneity of mutations in argininosuccinate synthetase causing human citrullinemia

Citrullinemia is an autosomal recessive disease caused by deficiency of argininosuccinate synthetase. In order to characterize mutations, RNA was isolated from cultured fibroblasts from 13 unrelated patients with neonatal citrullinemia. Ten mutations were identified by sequencing of amplified cDNA. Seven single base missense mutations were identified: Gly14----Ser, Ser180----Asn, Arg157----His, Arg304----Trp, Gly324----Ser, Arg363----Trp, and Gly390----Arg. Six of these missense mutations involved conversion of a CpG dinucleotide in the sense strand to TpG or CpA, and six of the seven mutations alter a restriction enzyme site in the cDNA. Two mutations were observed in which the sequences encoded by a single exon (exon 7 or 13) were absent from the cDNA. One mutation is a G----C substitution in the last position of intron 15 resulting in splicing to a cryptic splice site within exon 16. There is extreme heterogeneity of mutations causing citrulinemia. This heterogeneity may prove typical for less common autosomal recessive human genetic diseases.     

Innervation of the incisors and periodontal ligament in several rodents: an immunohistochemical study of neurofilament protein and glia-specific S-100 protein

The present immunohistochemical study by use of antisera against neurofilament protein (NFP) and S-100 protein dealt with the innervation of the upper incisors and periodontal ligament in five species of rodents including the guinea pig, hamster, Mongolian gerbil (Meriones unguicularis), mouse and squirrel (Tamias sibiricus). The innervation pattern of the periodontal ligament and dental pulp in the incisors of five rodents was fundamentally identical to that in the rat, which we have previously demonstrated by the same method. The NFP-positive Ruffini-like corpuscles were concentrated in the middle region of the lingual periodontal ligament in all the species examined, suggesting that this particular arrangement of Ruffini-like corpuscles, possibly stretch receptors, was essential to the rodent incisor. The labial periodontal ligament, on the other hand, contained less numerous NFP-positive nerves, these terminating among collagen fibers as free endings. The gerbil and squirrel in particular possessed only a few nerve fibers in the labial periodontal ligament. It was thus presumed that the labial periodontal ligament might be less significant as a mechanoreceptive site than the lingual periodontal ligament. The NFP-positive pulpal nerves, beaded or smooth in shape, ran parallel to the tooth axis, but never extended to the odontoblastic layer; no subodontoblastic plexus was found in the incisors of any of the rodents. S-100-immunopositive nervous elements were distributed in the periodontal ligament and dental pulp of all the rodent species examined, showing a distribution pattern similar to the NFP-positive nerves. Only in the squirrel did odontoblasts show an intense S-100 immunoreactivity.     

Comparison of the separation of Candida albicans chromosome-sized DNA by pulsed-field gel electrophoresis techniques.

Pulsed-field gel electrophoresis techniques were used to study chromosome-sized DNA molecules of C. albicans. Chromosome-sized DNA of two strains of Candida albicans has been resolved into 8 bands by orthogonal-field-alternation gel electrophoresis (OFAGE). Six bands were observed in chromosomal preparations of C. albicans using field-inversion gel electrophoresis (FIGE). Differences in the electrophoretic mobilities of bands of the strains of C. albicans examined suggests that chromosome-length polymorphisms exist and make it difficult to correlate the banding patterns among strains. These correlations were facilitated, however, by assignment of C. albicans chromosomes by hybridization using a collection of cloned DNA probes specific for each of the 8 observed bands. Southern blotting showed that the 6 FIGE bands consisted of 4 singlets and 2 comigrating doublets, accounting for the 8 bands observed by OFAGE analysis. The agreement between OFAGE and FIGE analysis suggests that the C. albicans haploid genome contains a minimum of 8 chromosomes.     

Metabolism of exogenous galactosylceramide in the twitcher mouse brain

The in vivo metabolism of galactosylceramide (gal-cer) in normal mice and in twitcher mice, a model of human GLD, was examined following intracerebral administration of gal-cer containing [1-¹⁴C]stearic acid. In normal mice, gal-cer was hydrolyzed to ceramide within 6 hours and ceramide was hydrolyzed to sphingosine and fatty acid. Most of the released fatty acid was immediately incorporated into other lipids. About 75% of injected gal-cer was hydrolyzed 80 hours after the injection, while in the twitcher mouse, only 17% of gal-cer was hydrolyzed. These results show that degradation of gal-cer is impaired in the twitcher mouse brain, but contradict to the fact that there was no evidence of any accumulation of gal-cer in the brain. This discrepancy may be due to the different sorting routes of biosynthesized and exogenously-administered gal-cer in the mouse brain. Most of the biosynthesized gal-cer is incorporated into myelin, while the injected gal-cer is incorporated into lysosomes.