Evidence for the presence of a histaminergic neuron system in the rat brain: an immunohistochemical analysis

The binding of calcium antagonists in the rat hippocampal formation was studied using autoradiography. Hippocampal slices were labeled in vitro with [3H]PN 200-110. High densities of binding sites for calcium antagonists were found in the molecular layer of the dentate gyrus and in the CA3 subfield of the hippocampus. After ablation of the granule cells by local injection of colchicine a marked decrease in the number of [3H]PN 200-110 binding sites density was observed on these areas, while binding to other parts of the hippocampal formation and brain was spared. These results strongly suggest the localization of high densities of calcium channels to the granule cells of the dentate gyrus.     

Complete Nucleotide Sequence and Phylogenetic Analyses of Hepatitis B Virus Isolated from Two Pileated Gibbons

Virus-like particles (VLPs, named HmTV1-17), about 40nm in diameter were found in the violet root rot fungus Helicobasidium mompa Tanaka strain No. 17, which had been isolated from an apple tree. Purified preparations of HmTV1-17 contained two species of double-stranded RNA (dsRNA), designated 17L and 17S. cDNAs were constructed from HmTV1-17 genomic dsRNAs purified using CF-11 cellulose column chromatography. The sequences of 17L and 17S cDNA comprised 5207 and 2096bp, respectively. Although 17S has no large open reading flame (ORF) on either strand, 17L has two large overlapping ORFs. The 5 located ORF1 encodes the coat protein (CP, 788 amino acids), whereas the gene product of ORF2, which is in the –1 frame relative to ORF1, shows the typical features of a RNA dependent RNA polymerase (RDRP, 845 amino acids). Phylogenetic analysis based on RDRP showed that HmTV1-17 is closely related to Sphaeropsis sapinea SsRV1, a member of the genus Totivirus from filamentous fungus S. sapinea.     

Effect of low calcium diet on the ultrastructure of the rat parathyroid gland

Young female rats were fed with normal (1.18%) or low (0.05%) calcium diet for 3, 7, 15 or 30 days. The morphology of the parathyroid glands was studied together with serum calcium, parathyroid hormone (PTH), calcitonin and bone mineral density (BMD). As compared to the animals fed with the normal calcium diet, BMD of whole body of the rats fed with the low calcium diet was significantly decreased, whereas the serum PTH level was increased. The parathyroid glands in the rats fed with the low calcium diet were markedly enlarged. In the parathyroid chief cells of the rats fed with the low calcium diet, the Golgi complexes and the cisternae of the granular endoplasmic reticulum were well developed, while the large granules and large vacuolar bodies decreased. Some secretory granules located near the plasma membrane. A proportionally larger increase of the cytoplasm was estimated in the rats fed with the low calcium diet for three and seven days. Enlargement of the cytoplasm and rather frequent mitoses of the chief cells were observed in the rats fed with the low calcium diet for 15 and 30 days. These findings suggest that the rapid bone loss in young rats induced by the low calcium diet is essentially due to stimulated activity of the parathyroid gland. The stimulated gland may be a result of hypertrophy at the early stage and a combination of hypertrophy and hyperplasia at the later stage of calcium deficiency.     

Raft.1, a monoclonal antibody raised against the raft microdomain,recognizes G-protein beta1 and 2, which assemble near nucleus after shiga toxin binding to human renal cell line

SUMMARY: Raft microdomains are glycolipid-enriched microdomain scaffolding molecules involved in signal transduction. The binding of Shiga toxin to globotriaosyl ceramide in raft microdomains of the human renal tubular cell line ACHN causes temporal activation of Src-kinase Yes. To study the downstream signaling mechanism proceeding to the activation of Yes, we raised monoclonal antibodies (MAbs) against raft microdomains. The MAbs were screened on the basis of, first, binding to raft microdomains with dot-blot immunostaining, second, intracellular localization of the epitope by flowcytometry after permeabilization, and third, translocation of the antigen molecules after Stx treatment by immunohistochemical staining. Raft.1 MAb bound to the molecules that accumulated to the particular region near the nucleus after Stx treatment. Two-dimensional Western blotting and matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis revealed that the antigen molecule is GTP binding protein beta subunits 1 and 2 (Gbeta1 and 2). That Raft.1 recognized Gbeta1 and 2 was further confirmed by the reactivity to recombinant Gbeta1 and 2 proteins. To our knowledge, this is the first report of production of a MAb recognizing Gbeta1 and 2. Because Gbeta1 and 2 are highly conserved all through organisms and are deeply involved in signal transduction, Raft.1 is expected to be utilized frequently in research.     

Typology of the arteries in the human scalenus region, with special reference to the accessory ascending cervical artery

The accessory ascending cervical artery (Murakami et al., 1996), which arises from the subclavian artery and ascends between the scalenus anterior and medius muscles, was studied in 87 Japanese adult cadavers (174 sides), with special attention being given to its origin, distribution, and relationship to other arteries at the cervical or scalenus region. In 154 sides (88.5%), the accessory ascending cervical artery was found to originate from the subclavian artery behind the scalenus anterior muscle, and to branch out to the scalenus anterior and medius muscles as well as those entering the 5th and 6th intervertebral foramens along the 6th and 7th cervical nerves. This artery arose independently in 105 sides. The accessory ascending cervical artery issued off or formed a common trunk with the transverse cervical artery and/or costocervical trunk in 49 sides. In cases lacking the accessory ascending cervical artery, it was usually compensated for by the costocervial trunk and/or transverse cervical artery (18 sides). Common trunk formation with the vertebral, internal thoracic, or suprascapular arteries was not observed. The authors suggest that the accessory ascending cervical artery, the transverse cervical artery, and the costocervical trunk should be grouped into one arterial system, a system that may be a remnant of the precostal longitudinal anastomoses of intersegmental arteries of the dorsal aorta behind the scalenus anterior muscle.     

Restriction endonuclease digest patterns of chromosomal DNA from group B beta-haemolytic streptococci.

Scanning densitometry and computer-assisted numerical analysis were used to examine restriction endonuclease digest patterns (RDPs) of chromosomal DNA from 26 infecting strains and 44 vaginal isolates of group B beta-haemolytic streptococci (GBS). At the 95% similarity level, HindIII RDPs of serotype Ia and III strains clustered into four and three RDP types, respectively. Nine of 10 strains from neonates with early-onset septicaemia belonged to two particular RDP types (Ia-3 and III-3). In contrast, serotype III GBS strains from meningitis cases were not characterised by particular RDP types. Associations between RDPs and certain phenotypic characteristics were also found.     

Detection of genetic alterations in advanced prostate cancer by comparative genomic hybridization

In this study, we examined nine cases of advanced Japanese prostate cancer by comparative genomic hybridization (CGH) to detect chromosomal imbalances across the entire genome and to identify several new regions likely to contain genes important to the development and progression of this disease. These cases had been previously examined for numerical chromosomal aberrations by fluorescence in situ hybridization (FISH). By CGH, the following regions were found to be over-represented (gains), with fluorescence ratio values higher than the threshold: 4p, 6p, 8q, 11q, 12q, 15q, 16p, 17q, 20, and 21 (>4 cases); underrepresentation (losses) involved: 1q, 4q, 5q, 6q, 13q, 14q, and 22 (>4 cases). The shortest regions of overlap (SRO) of gains were noted at 8q24.1 through q24.3, 12q23, and 17q23 through q24 (>5 cases). The SRO of losses were seen at 5q14 through q21, 6q16.1 through q21, 13q21.3 through q22, and 14q21 (>5 cases). Notably, the gain of chromosomes 8 and 12 by CGH was in agreement with the FISH data, suggesting that the gain of chromosomes 8 and 12 may play an important role in prostate carcinogenesis. The genes on the SRO regions were also discussed in relation to oncogenes and bone metastases.