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Biomedical subjects

Y Toyoda

Publications and source records attributed to Y Toyoda.

At least 19 recordsLinked to original sources

Tetraparental mice reveal complex cellular interactions of the mutant, autoimmunity-inducing lpr gene.

Lpr is a murine recessive autosomal mutant gene that, in conjunction with the background genes of different inbred mouse strains, produces varied syndromes of lymphadenopathy and autoantibody production. We have investigated the cellular basis of the lpr determined phenomena by constructing allophenic (tetraparental) chimeras of MRL/Mp-lpr/lpr combined with C57BL/6 and MRL/+ combined with C57BL/6-lpr/lpr. The appearance of lymphadenopathy, lpr atypical T cells, and autoantibodies was found to depend on the relative amounts +/+ and lpr tissue. The failure of these three abnormalities to correlate in certain animals suggested different pathways of lpr gene action. Furthermore, in the lpr predominant animals, both the lymphadenopathy and autoantibodies were essentially entirely contributed by the lpr donor. The expression of the lpr gene within the same cells expressing the MRL or B6 background determined the pathologic and serologic manifestations characteristic of the disease of that lpr strain, despite the presence of +/+ cells of the other background in the same individual. The lpr syndrome thus depends on the intracellular interaction of the lpr and background genes, and on positive signalling between different lpr cell populations.

Animals

Inhibition of mutagenesis by p-aminobenzoic acid as a nitrite scavenger.

Nitrite treatment enhances the direct-acting mutagenicity of various foodstuffs (e.g., chicken, bloater, the soybean flour 'kinako', and Ban-Ban-Chi sauce) on Salmonella typhimurium TA100. p-Aminobenzoic acid (PABA) and glutathione (GSH) reduced this mutagenicity; on the other hand, thioproline (TPRO) increased it. PABA seemed more effective than TPRO in scavenging nitrite ion. In analysis of the reactions of PABA and sodium nitrite under acidic conditions (pH 3.0), p-hydroxybenzoic acid (PHBA) was identified as a major reaction product. The reaction seems to involve two steps, diazotization and diazonium substitution. PHBA was not mutagenic to four strains (TA97, TA98, TA100 and TA102) of S. typhimurium with or without S9 mix.

4-Aminobenzoic Acid

Acute and late disease induced by murine coronavirus, strain JHM, in a series of recombinant inbred strains between BALB/cHeA and STS/A mice.

To examine the genetic control of acute and late disease induced by a murine coronavirus, strain JHM (JHMV), BALB/cHeA, STS/A, F1 hybrids and 13 recombinant inbred (RI) strains between BALB/cHeA and STS/A mouse strains were inoculated intracerebrally with 100 pfu of JHMV. All the BALB/cHeA mice died within 2 weeks from acute encephalitis. In contrast, STS/A mice were shown to be partially resistant, with a mortality rate of 30%, longer survival times and lower rates of viral production. The mortality rates, survival times and viral titers of F1 hybrids and the RI strains varied, suggesting involvement of multiple genes. STS/A, F1 hybrid and RI mice surviving the acute infection occasionally developed severe paraparesis about 1 month post-infection. In these mice, vacuolar degeneration, astrocytosis, the absence of perivascular cuffing and minimal demyelination were found in the central nervous system. No infectious virus could be recovered from these mice. Although the paralysis of delayed onset was limited to STS/A, F1 hybrid and eight of the 13 RI strains, the incidence varied significantly among the RI strains. These results may suggest that JHMV-induced late disease is also under multifactorial control. The pathogenesis of JHMV infection is discussed.

Acute Disease

Production of normal mice from oocytes fertilized and developed without zonae pellucidae.

Mouse oocytes, freed from zonae pellucidae, were inseminated and cultured in vitro. When inseminated with a modest concentration of spermatozoa (approximately 5/microliter), 79% of 151 oocytes were fertilized normally, of which the majority developed into blastocysts. When 177 blastocysts were transferred to 12 pseudopregnant foster mothers, 10 (83%) of the mothers became pregnant and 37 live pups were obtained. Although the overall rate of development of zona-free embryos was slightly inferior to that of zona-intact (control) embryos, these results nevertheless indicate that the oocyte and embryo are potentially capable of normal fertilization and development in the total absence of the native zona pellucida. Since the acrosome reaction is essential for successful sperm-oocyte fusion, zona-free oocytes must have been fertilized by spermatozoa which had undergone the reaction either in the medium or on the surface of the oocyte plasma membrane. Such acrosome reactions, which are not mediated by the native zona pellucida, can be called 'unphysiological' or 'spurious' but the spermatozoa with such reactions were certainly functional, as they produced healthy offspring.

Animals

Sexing of mouse preimplantation embryos by detection of Y chromosome-specific sequences using polymerase chain reaction.

Detection of genes known to be present on the mammalian Y chromosome was adapted for sexing mouse early embryos using the polymerase chain reaction (PCR) method. Sry and Zfy genes located in the sex-determining region of the Y chromosome were chosen for Y-specific target sequences, and DXNds3 sequence on the X chromosome was chosen for control. The two-step PCR method using two pairs of primers for each of the target sequences was employed for detecting the sequences. When DNAs of male and female mice were amplified with these primers, male-specific fragments were detected even in DNAs that were equivalent in amount to two cells. Mouse embryos at the two-cell stage were separated into two individual blastomeres, and one blastomere was karyotyped at the second cleavage. The remaining blastomere was subjected to PCR amplification immediately or after having been cultured for 48 h up to the morula stage. The Sry and Zfy sequences were detected in about half the embryos; detection of the Sry and Zfy sequences corresponded exactly to the presence of the Y chromosome, except in one sample of male morula in which embryos may have been lost before the PCR amplification. It is concluded that the sex of mouse preimplantation embryos can be accurately determined through detection of the Y-specific sequences using the two-step PCR method, even with the single blastomeres separated at the two-cell stage.

Animals

Maturation, fertilization, and development of dog oocytes in vitro.

Preovulatory oocytes were collected from ovaries of beagle bitches that had received superovulatory treatment. They were cultured in a modified Krebs-Ringer bicarbonate solution containing 10% fetal calf serum and 30 mg/L gentamicin sulfate for up to 72 h. About 32% of oocytes reached metaphase II by 72 h of culture. When these in vitro-matured oocytes were inseminated with ejaculated beagle spermatozoa that had been preincubated for 4 h, sperm penetration of the zona pellucida started about 1 h after insemination, and both male and female pronuclei were seen in the ooplasm at 8 h after insemination. At 18-20 h after insemination, oocytes were transferred to Whitten's medium and cultured for 76-78 h. The first cleavage was observed at 48 h after insemination, and 15 of 45 oocytes developed to the 2-8-cell stage. These results demonstrate that in vitro-matured canine oocytes can be fertilized and develop to the 8-cell stage in vitro.

Animals

Comparison of histone H1 kinase activity during meiotic maturation between two types of porcine oocytes matured in different media in vitro.

Histone H1 kinase (H1K) activity was assayed during meiotic maturation in porcine oocytes matured in a modified Krebs-Ringer bicarbonate solution (KRB) or in porcine follicular fluid (pFF) in vitro. Oocytes matured in KRB displayed lower male pronucleus formation ability, delayed first polar body emission, and a higher spontaneous activation rate than oocytes matured in pFF. In oocytes matured in pFF, H1K activity was low at the germinal vesicle stage and increased about 8-fold at first and second metaphases, with a transient depression at first anaphase and telophase. The H1K activity at second metaphase in oocytes matured in KRB was significantly lower than that in oocytes matured in pFF. These results suggest that the maturation medium used influences the fluctuation pattern of H1K activity and the biological characteristics of porcine oocytes cultured in vitro.

Animals

Effect of a partial deletion of Y chromosome on in vitro fertilizing ability of mouse spermatozoa.

The effect of a partial deletion of Y chromosome on sperm fertilizing ability was investigated through an in vitro fertilization technique. Epididymal spermatozoa of a congenic line, B10.BR-Ydel, which is characterized by a high incidence of abnormal spermatozoa, revealed a significantly lower in vitro fertilization rate (22%) than that (79%) of its control strain (B10.BR/SgSn), which has a normal-sized Y chromosome. Incidence of capacitated spermatozoa as determined by chlortetracycline fluorescence assay was significantly lower in B10.BR-Ydel than in B10.BR/SgSn spermatozoa. The fertilization rate was significantly improved when B10.BR-Ydel spermatozoa were separated from the supernatant of sperm suspension by Percoll gradient centrifugation. A reconstitution experiment revealed that the B10.BR-Ydel spermatozoa were more sensitive to the inhibitory effect of the supernatant than B10.BR/SgSn spermatozoa. Spermatozoa from F1 (C57BL/6N male x B10.BR-Ydel female) males showed higher fertilization rates than those from F1 (B10.BR.Ydel male x C57BL/6N female) males. These observations suggest that not only the morphology but also the fertilizing ability of spermatozoa is directly related to partial deletion of Y chromosome.

Animals

[Effect of tazanolast on platelet activating factor-induced airway hyperresponsiveness in guinea pigs].

To determine whether tazanolast inhibits airway hyperresponsiveness, we studied the effect of this drug on platelet activating factor (PAF)-induced airway hyperresponsiveness in guinea pigs. Inhalation of PAF (1 microgram/ml) caused significant airway hyperresponsiveness to acetylcholine (p < 0.01) or histamine (p < 0.05). Pretreatment with tazanolast (30-300 mg/kg) produced a dose-dependent inhibition of airway hyperresponsiveness induced by PAF inhalation, and significant inhibition (p < 0.05) was obtained with the drug (300 mg/kg). Aspirin also inhibited PAF-induced airway hyperresponsiveness, while tranilast produced hardly any inhibition. From these results, it is suggested that tazanolast is effective in inhibiting airway hyperresponsiveness.

Animals

Long-term results of knee synovectomy in pigmented villonodular synovitis.

1. Pigmented villonodular synovitis (PVS) commonly affects the knee as a monoarticular proliferative process. The etiology is not understood. 2. Partial synovectomy for localized PVS and wide synovectomy for diffuse PVS institute a favorable result, despite a rather high rate of recurrence at long-term follow-up. 3. Cases that involve bony erosion have a low incidence of recurrence but present an unsatisfactory prognosis even after curettage and wide synovectomy (postoperative progress of osteoarthritic changes).

Adult