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Biomedical subjects

M Shinagawa

Publications and source records attributed to M Shinagawa.

At least 109 records · Page 6Linked to original sources

Molecular epidemiology of adenoviral conjunctivitis in Sapporo, Japan, and Manila, the Philippines.

DNA restriction endonuclease analysis was carried out on 48 adenovirus isolates from the conjunctival swabs of patients with acute conjunctivitis. Samples were obtained in Sapporo, Japan, and Manila, the Philippines, from October 1983 to October 1984. We identified new genomic variants among the isolates of Ad3 in Sapporo, and Ad4 and Ad11 in Manila, and these were designated as Ad3f, Ad4b, Ad11a and Ad11b, respectively. Ad7 isolates in Manila were identical to Ad7b reported by Wadell et al. In Sapporo, no Ad7 was isolated during the period of this study. One isolate in Sapporo, which was neutralized by both Ad34 and Ad35 antisera, showed identical restriction patterns to Ad35 prototype strain (Holden) with SmaI, BamHI and HpaI. However, with HindIII, the pattern was identical to that of Ad34 prototype strain (Compton). This isolate was tentatively designated as Ad34/35. Ad8 isolates in Sapporo were identical to Ad8B detected there from 1977 to 1981, while Ad8 isolates in Manila were prototype (Trim) strains. In both cities Ad19 isolates were all identical to Ad19a and Ad37 isolates were all prototype (GW). Genomic variants of Ad isolates in both cities are as follows: (Table: see text).

Adenoviridae Infections↗

Bilateral sleeve lobectomy for metachronous multiple primary lung cancer.

In a patient with metachronous multiple primary lung cancer, bilateral lobectomy was performed, using bronchoplastic procedures. This case may be one of very few such cases reported in the literature. A 56-year-old man with squamous cell carcinoma was surgically treated for lung cancer. At the first operation, right upper lobectomy with wedge resection of the right main bronchus was performed, as the tumor occupied the orifice of the right main bronchus. Six months later, re-operation for stricture at the anastomotic line was done because of granulation. By means of sleeve resection of the strictured right main bronchus, the airway was reconstructed. The patient remained well for five years, then a similar cancer at the orifice of the left lower lobe and bulging into the left main bronchus became evident. Left lower sleeve lobectomy was done for the second primary cancer. The postoperative course was uneventful and he is well with no signs of recurrence 6 years and 10 months after the first operation and 19 months after the second sleeve lobectomy.

Carcinoma, Squamous Cell↗

Generation of packaging-defective DNA molecules of equine adenovirus.

Equine adenovirus (EAd) DNA prepared from infected bovine kidney (MDBK) cells contained additional sequences of about 100 to 700 bp at the left-hand end of the genome. These aberrant viral genomes were produced even after the first passage of the wild type EAd in MDBK cells and their relative amounts did not change significantly during serial passage. The left terminal fragments of two defective viral DNAs were cloned into the plasmid vector pBR322 and the nucleotide sequences of their terminal regions were analyzed. The data indicate that one viral DNA contained a duplication of the inverted terminal repetition (ITR) and the other contained 270 bp of additional sequences derived from the right-terminal region of EAd genome added to the left-terminal, ITR. While the former DNA was packaged into virions, the latter was not, presumable due to the alteration of the distance from the left terminus to the putative DNA packaging signal, reported to be located between 290 and 390 bp (Hammarskjold and Winberg, 1980). The possible mechanism for the generation of these defective DNAs is discussed.

Adenoviridae↗

Chronic obstruction of the inferior vena cava with recurrent endometrial stromal sarcoma.

A 64-year-old woman presented with endometrial stromal sarcoma of the inferior vena cava. The tumor was removed along the venous wall, and the vena cava and the right renal vein were reconstructed with a ringed polytetrafluoroethylene (PTFE) graft. The patient remains well 9 months after the operation despite graft occlusion. Fifteen years earlier the patient had undergone a hysterectomy for endometrial stromal sarcoma. Histologically, the second tumor was identical with the endometrial stromal sarcoma removed with the uterus at the primary operation. To the best of our knowledge, this is the first report of a case of obstruction of the inferior vena cava by such a tumor.

Chronic Disease↗

Immunoreactivity of a synthetic pentadecapeptide corresponding to the N-terminal region of the scrapie prion protein.

A pentadecapeptide with an amino acid sequence corresponding to the amino-terminal region of the scrapie prion protein was synthesized. Immunization of a rabbit with the peptide conjugated with ovalbumin induced specific antibodies. The antibodies reacted with all three of the major polypeptides in a proteinase K-treated fraction obtained from brains of mice infected with the Obihiro strain of scrapie agent. Some peptides in the proteinase-untreated fraction also shared antigenicity with the three major polypeptides. Specific polypeptides were also detected by the antiserum in a fraction prepared from spleens, but only two of the three major polypeptides were found and the amounts of the polypeptides were less than in brain.

Animals↗

Purification of scrapie agent from infected animal brains and raising of antibodies to the purified fraction.

The fraction (P4) containing scrapie infectivity was obtained by treatment of scrapie-infected mouse brains with the detergent sarcosyl, differential centrifugation, and proteolytic enzyme digestion. Scrapie infectivity in the P4 fraction was purified 239-2,390 times with respect to protein. Similar fractions were also prepared from the brain of a sheep naturally infected with scrapie. Morphological observation of the P4 fractions revealed that the main components were unique rods of 3-5 X 60-200 nm, which resembled scrapie-associated fibrils (SAF) or prion rods. The P4 fractions formed three major broad bands of polypeptides with molecular weights (MWs) of about 24.5K, 21K, and 17K dalton (Kd) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and some low MW polypeptides were also present in the fraction. Rabbits immunized with this fraction prepared from mouse brains raised antibodies against the three major polypeptides.

Animals↗

MDBK nuclear factor-binding site of various serotypes of adenovirus DNA.

A fraction with the ability to bind the terminal fragment of equine adenovirus (EAd) DNA was prepared from MDBK cell nuclei. The fraction (MDBK nuclear factor) bound to the terminal fragment of all human and animal adenovirus DNAs examined except avian adenovirus EDS-76. However, the terminal fragments of two animal adenoviruses, EAd and bovine adenovirus type 3 (BAd3), showed higher affinity for the nuclear factor than the others. The MDBK nuclear factor-binding site determined by footprinting analysis was the sequence located between nucleotides 22 and 46 in EAd, between 36 and 53 in canine adenovirus type 2, and between 20 and 46 in BAd3, counting from the terminus. The respective binding site contained a sequence resembling the consensus sequence. The binding site of Ad4 DNA was not within the inverted terminal repetition, but was located at least 550 base pairs apart from the terminus.

Adenoviridae↗

Conservation of DNA sequences for plasmid-mediated citrate utilization within the enterobacteria.

Southern blot DNA-DNA hybridization experiments with a cloned Cit+ DNA fragment as a probe showed that the plasmid-mediated Cit+ determinants from four Cit plasmids (R726, pOH3001, pOH3035, and pOH30221) were all homologous. Sequences homologous to the plasmid-borne Cit+ gene were also found in total bacterial DNA isolated from Salmonella paratyphi B, Salmonella enteritidis, Salmonella typhimurium LT-2, Citrobacter freundii, ATCC 8090, Citrobacter amalonaticus ATCC 25405, Klebsiella pneumoniae I and IID 977, and Enterobacter aerogenes ATCC 13048. The DNA digest from C. amalonaticus ATCC 25405 contained a 1.4-kilobase BamHI-HincII DNA fragment that was strongly homologous with and identical in size to the plasmid Cit+ probe.

Base Sequence↗

[Light and electron microscopic observations of sheep scrapie].

Two Suffolk sheep diagnosed as scrapie clinically and epidemiologically were investigated light and electron microscopically. They were female and four years four months of age. Spongiform lesions were found in the gray and white matter of midbrain, pons, medulla oblongata, spinal cord and the cerebellar white matter as well as the cerebral gray and white matter. Ultrastructurally, the spongiform lesions were shown to be caused by vacuolation in neuronal perikarya, vacuolation and/or swelling of neuropil, dilatation of the periaxonal space, and splitting of the myelin sheath followed by the intramyelinic vacuolation. Vacuole in neuronal perikaryon was associated with the enlarged endoplasmic reticulum. Adjacent to the vacuole, enlargement of the rough endoplasmic reticulum and cisterns of Golgi apparatus was observed. The cytoplasmic processes found in the intraneuronal vacuoles were formed by the separation of the cytoplasm caused by the continuous dilatation of the endoplasmic reticulum and cisterns of Golgi apparatus. This indicated that the degeneration of the neuronal perikaryon due to such separation contributed to the formation of the intraneuronal vacuole. Vacuolation and swelling with disappearance of the organelles were also found in astro- and oligodendroglial perikarya. Splitting of myelin sheath and the intramyelinic vacuolation were regarded as primary changes. The latter was supported by the degeneration of oligodendroglias found in this study.

Animals↗

Characterization of scrapie agent isolated from sheep in Japan.

A pathogenic agent isolated in mice from the brain of a sheep affected by scrapie-like disease was characterized. The incubation period of the disease in the primary transmission from the sheep to mice was longer than in the secondary and the tertiary transmission in the same strain of mice. Progressive dilution of the inoculum caused prolongation of the incubation period. The infectivity of the agent in a 10% brain homogenate persisted, but decreased about 10(3) to 10(4) times after heating at 100 C for 30 min. Histological changes in the diseased mouse brains consisted of vacuolation of the nerve cells and spongiform degeneration in the gray matter of the central nervous system. Fine rod-shaped granulae with a length of 3 to 5 nm were observed within the swollen neuropil, axon, and perivascular astrocytic process. No serum antibodies against available mouse viruses, parainfluenza type 1 virus, lymphocytic choriomeningitis virus, and mouse reovirus type 3, were detected in any mice used in the experiments. These findings demonstrate that the disease of the sheep was the first case of scrapie in Japan.

Animals↗

Two distinctive transforming DNA regions on the canine adenovirus type 1 genome.

By transfecting cloned DNA fragments of canine adenovirus type 1, a novel transforming region of mammalian adenovirus was demonstrated inside the viral genome. Besides the left-end SmaI-D fragment (left 17.2%), an internally located SmaI-E fragment (17.2-28.7%) of canine adenovirus type 1 was found to transform primary rat kidney cells.

Adenoviruses, Canine↗

Polypeptide involved in the Escherichia coli plasmid-mediated citrate transport system.

A genetic determinant conferring on Escherichia coli the ability to utilize citrate as a sole source of carbon and energy was subcloned into pBR322 from a naturally occurring, citrate utilization (Cit+) plasmid, pOH30221, and was localized to a 1.6-kilobase region by cloning and subsequent deletion analysis. Genetic expression of the Cit+ determinant in E. coli minicells revealed that the Cit+ determinant encoded a single, membrane-associated polypeptide with an apparent molecular weight of 35,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This polypeptide seemed not to be synthesized as a precursor with an amino-terminal signal sequence.

Base Sequence↗