Japanese Journal of Medical Science and Biology
Online ISSN : 1884-2828
Print ISSN : 0021-5112
ISSN-L : 0021-5112
Volume 47, Issue 3
Displaying 1-4 of 4 articles from this issue
  • Hiroshi URAKAMI, Mamoru TAKAHASHI, Michisato MURATA, Akira TAMURA
    1994 Volume 47 Issue 3 Pages 127-139
    Published: 1994
    Released on J-STAGE: March 19, 2010
    JOURNAL FREE ACCESS
    Leptotrombidium pallidum naturally infected with Rickettsia tsutsugamushi was reared and bred in our laboratory for several generations by brother and sister mating. The larvae and adults at the 8th and 9th generations were sectioned and observed by electron microscopy for analysis for the distribution of rickettsiae in the mites. The distribution densities of rickettsiae were markedly different among organs in each mite, but rickettsiae were seen in all the organs and tissues. Rickettsiae were distributed in the highest density in the salivary gland of larvae, and in the salivary gland, excretory bladder, epidermal layer, ovary and testis of adult mites. Only a few rickettsiae were recognized in the muscle of both larvae and adults. On the other hand, we found, in the infected family line used, a significant number of mites in which no rickettsiae were found by electron microscopy. The grouping of rickettsia-positive and -negative mites according to the parent family revealed that the efficiency of vertical transmission of rickettsia was different from one parent family to another. Thus, it became clear that a significant number of rickettsia-negative mites are produced in an infected family line.
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  • Koichi MAKIMURA, Somay Yamagata MURAYAMA, Hideyo YAMAGUCHI
    1994 Volume 47 Issue 3 Pages 141-156
    Published: 1994
    Released on J-STAGE: March 19, 2010
    JOURNAL FREE ACCESS
    A polymerase chain reaction (PCR) method capable of detecting both Aspergillus fumigatus infections, pulmonary non-fumigatus Aspergillus species (spp.) and Penicillium spp. from clinical specimens was established. The primer pair was designed on the basis of the sequence of the 18S-ribosomal RNA gene of A. fumigatus and P. notatum. A 385 by product was successfully amplified by this PCR method from all of 12 medically important Aspergillus spp. and Penicillium spp. (38 strains), but not from human, calf, Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), any of 14 medically important yeastlike fungal species tested (32 strains) including Candida al bicans, several non-albicans Candida and Saccharomyces cerevisiae, Cryptococcus neoformans, Mucor spp. or Pneumocystis carinii. This specificity was subsequently confirmed by Southern hybridization analysis. The established PCR can detect such a small amount as 1 pg of A, fumigatus DNA by staining the PCR product with ethidium bromide. With sputum specimens from clinically diagnosed aspergilloma patients, this PCR technique was demonstrated to be a more sensitive diagnostic method for Aspergillus infections than conventional culture techniques.
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  • Yumi KANEGAE, Miho MAKIMURA, Izumu SAITO
    1994 Volume 47 Issue 3 Pages 157-166
    Published: 1994
    Released on J-STAGE: March 19, 2010
    JOURNAL FREE ACCESS
    Recently, the adenovirus expression vector attracts much attention for the application to gene therapy and the method to purify and concentrate adenovirus without loss of infectivity has become very important, especially for animal experiments and gene therapy of humans. In this report, we show a simple and efficient method for purifying infectious adenovirus. The method consists of sequential centrifugation in CsCl step gradients without loss of infectivity and can be completed in one day. The method maintained the viral infectivity after 10-fold concentration and seemed to remove more than 99.9% of carried-over proteins. We showed also that the buffers for dialyzing the purified virions influenced the stability of infectivity. The buffers of 10 mM HEPES-1 mM EDTA -10% glycerol and PBS (-) -10% glycerol resulted in higher stability than did 10 mM HEPES-1 mM MgC12-10% glycerol. The method is may be useful in many applications of recombinant adenovirus.
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  • Kenzo KATO, Jing GUO, Fumiaki TAGUCHI, Osami DAIMARU, Masako TAJIMA, H ...
    1994 Volume 47 Issue 3 Pages 167-178
    Published: 1994
    Released on J-STAGE: March 19, 2010
    JOURNAL FREE ACCESS
    We examined the phylogenetical correlation between two types of JC virus (JCV) isolates, archetypes derived from the urine of nonimmunocompromised individuals and PML-types derived from the brain of patients with progressive multifocal leukoencephalopathy (PML) in Japan. A phylogenetic tree was constructed for eight JCV isolates, five PML-types obtained in this and previous studies and three representative archetypes, from DNA sequence data on the VP1 (major capsid protein) gene. The eight isolates were divided into two major groups, named subtypes MY and CY after the representative archetypal isolates. Four of five PML-type isolates belonged to subtype MY, and the other one to subtype CY.Isolates belonging to subtype MY were further divided into two groups; one group containing archetype MY and three PML-types and the other one containing archetype YI and a PML-type. These findings, together with those in our previous study that correlated various JCV isolates in the world provide evidence for the hypothesis that JCVs associated with PML may have been generated from archetypal JCVs persisting in the patients.
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