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Biomedical subjects

H Urabe

Publications and source records attributed to H Urabe.

At least 19 recordsLinked to original sources

Nucleotide sequence and transcriptional analysis of activator-regulator proteins for beta-lactamase in Streptomyces cacaoi.

The nucleotide sequence of the 2.7-kb DNA fragment upstream of the structural gene of beta-lactamase in Streptomyces cacaoi was determined. Computer-aided "FRAME" analysis revealed four possible open reading frames (ORFs), three in one direction and one in the opposite direction. One of them (ORF1, BlaA) encoded an activator-regulator protein whose deduced amino acid sequence was similar to that of other activator-regulator proteins in bacteria. Insertion of an 8-bp BamHI linker into the BlaA region decreased the beta-lactamase activity sharply, from 50 U to 1 U/ml. This protein (BlaA) was found to bind to the nucleotide sequence between the bla (beta-lactamase structural gene) and blaA genes. Another ORF (ORF2, BlaB) in the same orientation had a couple of amino acid sequences similar to that of pBR322 beta-lactamase. However, insertion of the 8-bp BamHI linker indicated that this ORF was functional as an activator-regulator but not as a beta-lactamase. Therefore, there were two activator-regulator proteins in the upstream region of the structural gene of the beta-lactamase. Nuclease S1 mapping predicted that transcription for the activator proteins commenced at the translational initiation codon or within a few nucleotides from the translational start site. Transcription was in the opposite direction to that of the beta-lactamase structural gene.

Amino Acid Sequence

Malignant Triton tumor. A case with protean histopathological patterns.

A case of malignant Triton tumor occurring in the leg of a 48-year-old woman with neurofibromatosis (von Recklinghausen's disease) is presented. The neoplasm was composed mainly of spindle-shaped cells forming interlacing fascicles with areas of massive necrosis. The important feature in this neoplasm was the presence of rhabdomyoblastic cells admixed with other cellular components. In addition, variable histologic features including solid, myxomatous, hemangiomatous, hemangiopericytomatous, epithelioid, and lymphomatous areas were identified. This variety has been recognized in malignant schwannomas, but not in malignant Triton tumors. There were discrete foci with each predominant histologic feature shown in a map of the neoplasm in which the distribution of those components is demonstrated based on the histology and immunohistochemistry. Satisfactory biopsy specimens are necessary for accurate diagnosis of malignant Triton tumor.

Actins

A new apparatus for hair regrowth in male-pattern baldness.

A newly devised apparatus (Scalp-Tension-Relaxer, STR) can efficiently promote hair regrowth in patients with male-pattern baldness. When this apparatus is applied, the scalp is pushed up to relieve tension on the vertex. The efficacy rate of hair regrowth in alopecia patients was 40%. An investigation into the basis for the hair regrowth caused by this apparatus was directed toward the changes in hemodynamics and skin temperature of the scalp. During and after use of this apparatus, subjects exhibited an increase both in the cutaneous blood flow rate (as determined by laser Doppler flowmetry) and in the cutaneous temperature (as determined by thermography).

Adult

Beta-lactamase expression in Streptomyces cacaoi.

Plasmids were prepared by inserting genomic DNA fragments from Streptomyces cacaoi within the mel gene of plasmid pIJ702. The inserted DNA fragments contain the beta-lactamase-encoding bla gene and upstream nucleotide sequences of various lengths. The transcription start point of bla was identified by nuclease S1 mapping. Upstream nucleotide sequences of sufficient lengths had an enhancing effect on beta-lactamase production by the Streptomyces host. The dot blot hybridization assay revealed that this effect was exerted at the transcriptional level. Experimental evidence strongly suggests that the underlying mechanism involves, at least in part, one or several trans-acting elements. In one of the constructs, in which the upstream nucleotide sequence was reduced to 0.3 kb, the bla promoter was present but the bla gene was expressed by readthrough from a promoter, possibly the mel promoter, of the pIJ702 vector.

Amino Acid Sequence

Cloning from Streptomyces cellulosae of the gene encoding beta-lactamase, a blue-dextran binding protein.

A beta-lactamase gene was cloned from Streptomyces cellulosae as a 2.3-kb DNA fragment using Streptomyces lividans 1326 and PIJ385 as a host-vector system. During the course of cloning, a part of the chromosomal DNA fragment cloned together with a part of the vector plasmid were deleted, indicating instability of this contiguous DNA region. The enzyme from the clone showed similar properties with respect to binding of blue dextran and isoelectric point to the enzyme from S. cellulosae. The cloned gene hybridized not only to DNA of S. cellulosae, the source of DNA, but also to DNAs of several Streptomyces species, irrespective of their formation of beta-lactamase. These results suggest that this gene may have homology to genes other than the one for beta-lactamase.

Blotting, Southern

Orientation of retinal in purple membrane determined by polarized Raman spectroscopy.

The orientation of the retinal molecule in the purple membrane was determined by polarized Raman spectroscopy for stacked purple membranes. The depolarization ratios of C = C stretching vibration mode were measured for three scattering geometries of purple membrane films. From the depolarization ratios we estimated the tilt angle of the transition dipole moment of retinal to the membrane normal and the rotational angle of the molecular plane along the transition dipole moment of retinal. The molecular plane of M intermediate was found to be almost perpendicular to the membrane plane. We confirmed that the tilt angle was 65 +/- 2 degrees for both bR and M intermediates.

Bacteriorhodopsins

Identification of the minimal replication region of the multicopy Streptomyces plasmid pSL1.

The 1.52-kb minimal replication origin of the 3.9-kb Streptomyces plasmid pSL1 was determined using a bifunctional derivative, pMCP44, of pSL1. Plasmids with linker insertions into the pSL1 part of pMCP44 were isolated from Escherichia coli. The sites of insertion were determined by restriction enzyme analysis and the ability of the mutant plasmids to replicate in S. lividans 66 was determined. All except one of the inserts in the 1.52-kb essential region inactivated replication. A 104-bp segment from this region could function as a replication origin in the presence of a helper plasmid containing a nonoverlapping pSL1 fragment. The sequence of this 104-bp fragment shows similarities to those of known plasmid replication origins.

Base Sequence

Characterization of the incompatibility region of Streptomyces plasmid pSL1.

The incompatibility region of the Streptomyces plasmid pSL1 was identified as a 240-bp segment, though some other function from the vector plasmid was also necessary. A 540-bp fragment including the 240-bp region was enough for full expression of incompatibility. Inserted mutation analysis led to a more detailed location of the region essential for replication.

Chromosome Mapping

A new concept of basal cell epitheliomas based on the three-dimensional growth pattern of the superficial multicentric type.

Little is known about the growth characteristics of basal cell epitheliomas (BCEs), largely because of the difficulty in studying these tumors, both in situ and in vitro. In this study, the authors report that a scanning electron microscopic examination of 2 cases of superficial BCEs reveals that the tumor proliferates centrifugally as a monolayer beneath the epidermis as well as over the surfaces of hair follicles, while eroding and replacing the original basal architecture. Behind the leading edge of this monolayer, the cells split away from the epidermis to form a round border resembling a carpet being rolled up. This border then develops into a mass lesion, which represents the tumor cell foci seen in routine paraffin sections. These findings have led the authors to propose a fundamentally different explanation for the pathogenesis of these tumors: BCEs are conditioned neoplasms that require contact with the epidermal or hair follicular epithelium in order to grow. This hypothesis could explain the behavior of BCEs as well as the clinical and pathologic varieties observed in clinical practice.

Aged

The elevation of the cytoplasmic calcium ions in vascular smooth muscle cells in SHR--measurement of the free calcium ions in single living cells by lasermicrofluorospectrometry.

We have developed an accurate and sensitive system for the measurement of cytoplasmic free calcium concentrations ([Ca++]i) of a single cell by using UV-laser and Indo-1. By this method, we made the first successful measurement of [Ca++]i of single living vascular smooth muscle cells. [Ca++]i in spontaneously hypertensive rats was elevated and maintained after the 6th passage culture. However, [Ca++]i in Goldblatt hypertensive rats was not elevated. Thus, these results suggest that the maintenance of high [Ca++]i levels of vascular smooth muscle cells in spontaneously hypertensive rats is genetically regulated and that it is one of the mechanisms for hypertension.

Aging

Hypoplasty of the breast due to x-ray irradiation.

We treated five women with hypoplasty of the breast induced by radiation. Only one of these women underwent mammaplasty with the use of a latissimus dorsi muscle flap and prosthesis. Hypoplasty of the breast is considered a result of imprudent utilization of x-ray irradiation of young patients with benign skin diseases. To prevent underdevelopment and hypoplasty, attention must be directed to the risk involved in radiation therapy for benign diseases.

Adolescent