Biomedical subjects
Publications and source records attributed to T Ono.
Induction of chromosome aberrations was investigated in cultured Chinese hamster cells treated with pyrimethamine. Pyrimethamine without metabolic activation strongly induced structural chromosome aberrations in a dose-dependent manner. Aberrant metaphase cells occurred at a frequency of 80%, when cells were treated at 1.6 microgram/ml for 48 h.
The boundary element method was applied to 3-dimensionally reconstructed monkey brain to localize a current source generator (dipole) of somatosensory evoked potentials (SEPs) recorded from epidural electrodes under assumption that a brain is electrically homogeneous and is bounded by a high resistant substance (skull). Absolute mean deviation of estimated dipoles from actual dipoles artificially generated by electrical stimulation at known coordinates in the brain were within 1-3 mm. The estimated dipole computed from SEPs by right median nerve stimulation at latency of P10-N10 waves was located in area 3b of the somatosensory hand area in monkey. Our data suggest that this technique of locating dipoles by the boundary element method applied to a reconstructed brain allows accurate determination of the 3-dimensional coordinates of a dipole.
Current source generators (dipoles) of somatosensory evoked potentials (SEPs) in response to electrical stimulation of the median nerve in anesthetized monkeys were localized by the dipole tracing method (DT) which can approximate surface potential distributions to an equivalent dipole. The locations and latencies of dipoles estimated by DT were confirmed by recording multiple unit activities from the parietal cortices of the same monkeys investigated by DT. Loci of generators for each SEP component were: P10 and N10, area 3b; P12, areas 1 and 2; P18, area 5. The results indicate that DT could trace the successive movement of generators of SEPs from the primary somatosensory area to area 5, which has been suggested in neuroanatomical studies to be part of the hierarchical sequence of somatosensory processes.
We report here on two siblings who show no clinical manifestations except for slight cutaneous photosensitivity and cutaneous pigmentation but have biochemical characteristics of Cockayne syndrome (CS). Fibroblasts derived from the patients (Kps2 and Kps3) were 3-4 times more sensitive to UV than normal cells. Although unscheduled DNA synthesis (UDS) in these cells was at a normal level, recovery of RNA synthesis (RRS) after UV irradiation was severely depressed. Microinjection of bacteriophage T4 endonuclease V into the cells corrected RRS after UV irradiation to a level near normal. These results indicate that DNA repair of cyclobutane-type pyrimidine dimers is impaired in the cells and the biochemical characteristics are similar to those of CS cells. However, cell fusion complementation tests with CS group A and B cells resulted in correction of RRS after UV irradiation. Cell fusion with XP group A, B, D, F and G cells also corrected RRS after UV irradiation, and microinjection of cell extracts prepared from Kps3 cells corrected UDS in XP group C and E cells, indicating that the patients do not belong to any complementation group of XP or CS. These results suggest that the patients have a new UV-sensitive syndrome with a biochemical phenotype of CS.
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A major role of DNA-methyltransferase (MTase) is thought to be maintaining the DNA methylation profile through DNA replication. However, previous surveys of mRNA distribution in different tissues by Northern-blot analysis have shown unexpectedly high levels of expression of DNA-MTase mRNA in adult mouse brain, which consists mostly of slowly proliferating glial and nonproliferating neuronal cells. In order to identify cells expressing the gene in the brain, we performed an in situ hybridization analysis of mature brain as well as whole embryos of different ages. In addition to various embryonic tissues with active cell proliferation such as the ventricular neurogenic layer, hair follicle epithelia, thymus and epithelia of the base of intestinal villi, almost all mature neurons in brain of adult and even aged mice expressed DNA-MTase mRNA at substantial levels. No significant expression of the gene was detected in the white matter. These findings suggest some neuron-specific biological function of DNA methylation, unrelated to DNA replication.
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1. The relationship of the decreased caecal urease activity by dietary penicillin to nitrogen utilisation was assessed in chickens fed a low protein diet plus urea. 2. Dietary penicillin at 20 and 100 mg/kg decreased anaerobic bacteria counts, urease activity and ammonia concentration in caecal contents (P < 0.05, except for ammonia in the case of the 100 mg/kg penicillin diet). 3. The 20 mg/kg penicillin diets significantly increased the excretion of urea and total nitrogen (P < 0.05) and decreased ammonia excretion, and significantly reduced nitrogen retention (P < 0.05). The 100 mg/kg penicillin diet also resulted in similar but not significant changes, which tended to be less than those by the 20 mg/kg penicillin diet. 4. Ammonia, urea, glutamine and uric acid concentrations in blood, liver and kidney were unchanged by dietary penicillin. 5. It is concluded that caecal ammonia production from urea was closely correlated with nitrogen utilisation in chickens fed a low protein diet plus urea.
1. The effects of dietary penicillin on the urease activities of small intestine, large intestine, caecum and their contents, liver and kidney in chickens fed a diet containing 5 g/kg protein plus urea were examined. 2. About 0.88 of the total urease activity determined was observed in intestinal contents, of which 0.95 of the activity was accounted for by caecal contents, 0.05 by colo-rectal contents and none by small intestinal contents. Intestinal tissues (caecum included), liver and kidney accounted for 0.03, 0.06 and 0.02, respectively, of the total urease activity. 3. Dietary penicillin decreased urease activity to 0.17 in caecal contents and to 0.05 in colo-rectal contents of the corresponding control values (P < 0.01). The urease activity of caecal tissue was lowered by penicillin to half that of control activity (P < 0.05) but none of the activities of other tissues were affected. 4. It is concluded that, even when the urease activity is stimulated by dietary urea, 20 mg/kg dietary penicillin can strikingly lower it in the caecum, where most of the urease activity in the chicken body is to be found.
Synthetic oligonucleotide primers were used in the polymerase chain reaction (PCR) to amplify a sequence of the spaP gene, which encodes the surface protein antigen I/II of Streptococcus mutans. A DNA fragment of c. 192 bp was amplified from lysed S. mutans cells or isolated DNA. With S. mutans cells, the lower limit of detection was 4-40 cfu. With these primers, 13 reference and 50 clinical strains of S. mutans were identified. Amplification of the 192-bp product was not demonstrated when 41 strains of other streptococcal and non-streptococcal species were tested. The spaP gene PCR has potential for the rapid diagnosis of S. mutans infections.
Skin lesions in mycosis fungoides (MF) and cutaneous-type adult T-cell leukemia/lymphoma (c-ATL) are sometimes difficult to differentiate. Our previous investigations showed that the alpha-chain of interleukin-2 receptor (IL-2R alpha) secretion is enhanced in both acute and chronic type ATL. This finding suggested increased levels of IL-2R alpha in c-ATL that might differentiate it from MF. In this study, we report several observations on IL-2R alpha levels in MF and c-ATL. 1) Plasma levels of IL-2R alpha determined by enzyme-linked immunosorbent assay were higher in patients with c-ATL (tumor group, 17,571 +/- 5,919 U/ml; non-tumor group, 4,546 +/- 1,788 U/ml) than those of patients with MF (tumor group, 2,352 +/- 861 U/ml; plaque group, 530 +/- 174 U/ml). 2) The percentages of IL-2R alpha-positive cells in skin lesions of c-ATL were 30.5 +/- 13.4% in the non-tumor group and 62.7 +/- 12.0% in the tumor group, which were significantly higher than those of MF, plaque group (12.7 +/- 5.9%), and tumor group (3.4 +/- 2.3%). 3) Further, there was a significant correlation between plasma IL-2R alpha levels and the number of IL-2R alpha-positive cells in skin lesions in patients with c-ATL (r = 0.654, p < 0.0001) and MF (r = 0.537, p < 0.01) on an individual basis; whereas no increased IL-2R alpha levels in peripheral blood mononuclear cells were observed in patients with either MF or c-ATL. 4) Skin lesions of c-ATL had much higher levels of IL-2R alpha genomic RNA than those of MF in their initial stages when measured by DNase/reverse transcription/polymerase chain reaction and liquid hybridization analysis. These findings suggest that increased plasma IL-2R alpha levels in patients with either MF or c-ATL may be linked to cellular IL-2R alpha levels in the skin lesions, and thus reflect the difference between MF and c-ATL.
A boy with skin eruptions resembling varicella and specific for Langerhans cell histiocytosis (LCH) is reported. At his initial visit when he was four months old, vesiculopustular lesions were present over the entire body; these had first appeared on the third day post partus. Histopathological, immunohistochemical, and electron microscopical examination confirmed the Langerhans cell phenotype and Birbeck granules in the responsible cells. He also had hydronephrosis, recurrent fever, and cutaneous bacterial infections. His parents refused further medical treatment and he died of diarrhea with cachexia about two years later. LCH may present diagnostic difficulties by manifesting as a skin eruption which resembles varicella.
LEC rats develop an autosomal recessive hepatitis and subsequently liver cancer associated with copper accumulation in the liver similar to that of Wilson's disease. Using 71 backcross [(WKAH x LEC) x LEC] rats, linkage analysis of the hepatitis with the WD gene for Wilson's disease revealed identical segregation and no recombination event between these two genes. This result indicates that the WD gene is a prime candidate for the hts gene responsible for the hepatitis of LEC rats, and suggests that the hepatitis of LEC rats may be caused by a defect in a copper-transporting ATPase expressed in the liver.
Approximately 1% (4/500) of blood donors exhibited seropositivity for HTLV-II in South Vietnam, but none (0/500) did in North Vietnam. Further, all individuals seropositive for HTLV-II were intravenous drug abusers who were seronegative for HIV-1 and HTLV-I. These findings suggest that HTLV-II infection may be specifically prevalent in drug abusers in South Vietnam.
By establishing a unique screening method, we have isolated yeast mutants that die only after differentiating into cells with a mating projection, and some of them are also defective in Ca2+ signaling. The mutants were classified into five complementation groups, one of which we studied extensively. This mutation defines a new gene, designated MID1, which encodes an N-glycosylated, integral plasma membrane protein with 548 amino acid residues. The mid1-1 mutant has low Ca2+ uptake activity, loses viability after receiving mating pheromones, and escapes death when incubated with high concentrations of CaCl2. The MID1 gene is nonessential for vegetative growth. The efficiency of mating between MATa mid1-1 and MAT alpha mid1-1 cells is low. These results demonstrate that MID1 is required for Ca2+ influx and mating.