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Y Mukai

Publications and source records attributed to Y Mukai.

At least 55 records · Page 3Linked to original sources
PubMed1998

Studies on neurosteroids. VII. Determination of pregnenolone and its 3-stearate in rat brains using high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

An assay method for pregnenolone and its 3-stearate in rat brains has been developed using LC-atmospheric pressure chemical ionization (APCI) isotope dilution MS. The extraction of the rat brain homogenate containing deuterated internal standards (ISs) with organic solvent followed by silica gel mini-column chromatography gave two fractions containing pregnenolone and its 3-stearate together with the respective IS. Each fraction was derivatized into 3-acetate-20-methyloxime and 20-methyloxime, respectively, followed by silica gel mini-column chromatography to remove the excess reagents, and then each derivatized fraction was applied onto reversed-phase LC-APCI-MS. The method was applied to the determination of these steroids in the gray matter and olfactory bulbs of rat brains, which were divided into control and acute stressed specimens. Although pregnenolone in both regions of the rat brains increased more than five times after stress, its 3-stearate decreased after stress.

Animals↗
PubMed1998

Isolation and characterization of an invertase and its repressor genes from Schizosaccharomyces pombe.

PCR was used to isolate an invertase homolog gene from the fission yeast Schizosaccharomyces pombe. The cloned inv1(+) gene encodes a protein of 581 amino acids with 16 potential asparagine-linked glycosylation sites, and has 39% and 38% identity to the Schwanniomyces occidentalis and Saccharomyces cerevisiae SUC2 invertases. When the inv1(+) gene was disrupted, S. pombe strains lacked detectable invertase activity. This result showed that the inv1(+) gene encodes only one active invertase in S. pombe cells. The transcription of inv1(+) is repressed in the presence of glucose. The transcription of inv1(+) was not affected in cyr1Delta strain which lacks adenylate cyclase activity, unlike transcription of S. pombe fbp1(+) gene. We have identified an S. pombe gene (scr1(+)) that encodes a homolog of the Aspergillus nidulans CREA which is required for glucose repression of the glyconeogenic pathway. Although the deletion of scr1(+) did not influence the transcription of fbp1(+) gene, glucose repression of the inv1(+) gene was severely affected. These results showed that glucose repression of inv1(+) gene is dependent on scr1(+) gene, and S. pombe cAMP signalling pathway may not be essential for glucose repression of inv1(+) gene.

Amino Acid Sequence↗
PubMed1998

Elevated expression of differentiation inhibitory factor nm23 mRNA in monoblastic crisis of a patient with chronic myelogenous leukemia.

Differentiation inhibitory factor nm23 gene has been found to be expressed in high quantities in acute myelogenous leukemia (AML), especially in acute monocytic leukemia (AML-M5) and is suggested as a new prognostic factor in AML-M5. We report an example of elevated expression of nm23 mRNA in a patient with chronic myelogenous leukemia (CML) who developed monoblastic crisis. Relative levels of nm23-H1 and -H2 mRNA extracted from the patient's peripheral blood mononuclear cells and bone marrow mononuclear cells were measured by quantitative reverse transcriptase polymerase chain reaction. The level of nm23-H1 mRNA in CML cells at the chronic phase was as high as that in bone marrow cells from healthy volunteers. The mRNA level of nm23-H2 was slightly below the normal level. At blastic crisis, however, expression of both nm23-H1 and -H2 mRNA was elevated to about three to nine times of that at the chronic phase. Proliferated blastic cells were positive for non-specific esterase, and the serum lysozyme level was elevated and diagnosed as monoblastic crisis. The patient received combined chemotherapy but response was partial. These findings are compatible with our previous report that nm23 gene is overexpressed in monocytic leukemia.

Antigens, Neoplasm↗
PubMed1998

Dual rearrangement of immunoglobulin and T-cell receptor genes in a case of Philadelphia chromosome-positive acute leukemia.

A 48-year-old patient was admitted to our hospital for leukocytosis. The blast cells were positive for peroxidase and he was tentatively diagnosed as acute myeloid leukemia according to the French-American-British criteria. By flow cytometry, the bone marrow cells were positive for CD10, CD13, CD33 and HLA-DR, but two-color analysis revealed that most of the CD13- and CD33-positive cells did not express CD10. The marrow cells had Philadelphia chromosome with no additional abnormalities. Major bcr-abl fusion gene was observed by the reverse transcriptase-polymerase chain reaction method. Southern blot analysis disclosed rearrangement of both immunoglobulin heavy chain and T-cell receptor beta chain genes. He received combined chemotherapy for myeloid lineage and lymphoid lineage, but the response was quite poor. He died 64 days after admission due to pulmonary bleeding. Although the association of Ph1 with multilineage differentiation is unclear, our case has significant implication for further investigation of the relationship between Ph1-positive cells and lineage selection.

Acute Disease↗
PubMed1998

Dentinal tubule occlusion with lanthanum fluoride and powdered apatite glass ceramics in vitro.

To simulate hypersensitive dentin, the smear layer and dentinal plugs of bovine root dentin specimens were removed by immersion in 10% phosphoric acid, polishing with hydroxyapatite particles, and ultrasonic cleansing. The fluoride-tannic acid-lanthanum-apatite (FTLA) group was treated with acidulated phosphate fluoride (APF) containing tannic acid followed by rubbing with a paste of lanthanum chloride (LaCl3) and powdered apatite glass ceramics. The treated specimens were immersed in a remineralizing solution that mimics saliva for 6 weeks. The SEM observations revealed that the treated surfaces of the FTLA group were completely covered with fine spherical compounds and the dentinal tubules were occluded with plugs to a depth of about 3 microns. Fluoride and lanthanum were detected to a depth of over 20 microns by EPMA observation. After the remineralization, the surface of FTLA-treated specimen did not have any opened tubules and showed a remarkable increase in the number of fine spherical deposits in the dentinal tubules. These results suggest that the reaction products produced by sequential treatment with acidic fluoride and LaCl3 and powdered apatite glass ceramics are able to effectively occlude dentinal tubules.

Acidulated Phosphate Fluoride↗
PubMed1997

Clonal integration and expression of human T-cell lymphotropic virus type I in carriers detected by polymerase chain reaction and inverse PCR.

Adult T-cell leukemia (ATL) is a neoplasm of mature helper (CD4) T lymphocytes, and human T-cell lymphotropic virus type-I (HTLV-I) has been suggested to be the causative virus of ATL. HTLV-I integrates its proviruses into random sites in host chromosomal DNA. Clonal integration has been observed in patients with ATL, including smoldering, chronic, and acute states. However, random and/or polyclonal integration has only been reported in a few asymptomatic HTLV-I carriers. To clarify the clonality of HTLV-I-infected cells in carriers, we used an inverse polymerase chain reaction (IPCR), which is more sensitive than Southern blot analysis. We used the peripheral blood momonuclear cells (PBMC) from 16 asymptomatic carriers and the separated CD4-positive cells. No cases showed either a monoclonal or polyclonal integration of the HTLV-I provirus by Southern blot. But, using IPCR, 7 of 16 cases showed either mono- or oligoclonal integration. In addition, the populations of clonal provirus in the total PBMC were frequently different from those in the CD4-positive cells. Three cases showed expression of HTLV-I tax/rex mRNA in the total PBMC, but no such expression was found in CD4-positive cells. In this study, an unexpected frequency of clonal HTLV-I provirus DNA was observed in HTLV-I carriers. These findings indicate that the clonal but nonmalignant proliferation of HTLV-I-infected cells already occurs even in HTLV-I carriers, and therefore that some other step is necessary to induce malignant proliferation.

Adolescent↗
PubMed1997

The role of cysteine residues in the homeodomain protein Mat alpha 2 in mating-type control of Saccharomyces cerevisiae.

The Mat alpha 2 homeodomain protein plays a pivotal role in the control of cell type in Saccharomyces cerevisiae. The homeodomain in the C-terminal region of Mat alpha 2 functions as a DNA-binding domain and the N-terminal region, containing two cysteine residues at positions 33 and 34, is thought to be involved in formation of Mat alpha 2 homodimers via disulfide bonds. mat alpha 2 mutants, isolated in a previous study, in which haploid-specific genes cannot be repressed by the Matal-Mat alpha 2 heterodimer but a-specific genes can be repressed by the Mat alpha 2 homodimer, were found to produce mutant Mat alpha 2 with a substitution of tyrosine or phenylalanine for Cys33. To clarify the role of Cys33 and Cys34 in the Mat alpha 2 protein, we generated several mat alpha 2 mutants by site-directed mutagenesis which had serine residues in place of these Cys residues. Transforming MATa cells with plasmids carrying these mat alpha 2 (MAT alpha 1+) mutations rendered transformants unable to mate. Northern blot analysis revealed that transcription of the a-specific gene STE2 and the haploid-specific locus RME1 in these transformants is repressed to the same level as in wild-type MATa/MAT alpha cells. We concluded that neither Cys33 nor Cys34 is required for repression of a-specific genes by the Mat alpha 2 homodimer or of haploid-specific genes by the Matal-Mat alpha 2 heterodimer, and therefore suggest that Mat alpha 2 homodimer formation in vivo is not mediated by disulfide linkage.

Cysteine↗
PubMed1997

A complex arrangement of genes at a starch branching enzyme I locus in the D-genome donor of wheat.

Genomic DNA fragments from Triticum tauschii (D-genome donor to wheat) carrying starch branching enzyme I (SBE I) type genes have been characterized. One fragment contains one complete gene and two partial genes in 16 kb of DNA. One of the partial genes is oriented in the opposite strand to the other two. The gene that is complete was sequenced. Its structure corresponds closely to that of rice in that exons 3-8 are retained at similar sizes and spacings. A cDNA closely corresponding to the complete gene was isolated and characterized; it codes for a putative protein that represents a novel type of SBE I, as it is shorter at the 3' end than the forms reported so far in other plants. A second genomic fragment contains a different SBE I gene. There appear to be approximately 10 copies of SBE I type genes in wheat (approximately 5 in T. tauschii) and most of them have been assigned to group 7 chromosomes. In situ hybridization indicates that a major locus for the genes is located at the distal end of the short arm of chromosome 7D.

1,4-alpha-Glucan Branching Enzyme↗
PubMed1997

Identification of individual barley chromosomes based on repetitive sequences: conservative distribution of Afa-family repetitive sequences on the chromosomes of barley and wheat.

The Afa-family repetitive sequences were isolated from barley (Hordeum vulgare, 2n = 14) and cloned as pHvA14. This sequence distinguished each barely chromosome by in situ hybridization. Double color fluorescence in situ hybridization using pHvA14 and 5S rDNA or HvRT-family sequence (subtelomeric sequence of barley) allocated individual barley chromosomes showing a specific pattern of pHvA14 to chromosome 1H to 7H. As the case of the D genome chromosomes of Aegilops squarrosa and common wheat (Triticum aestivum) hybridized by its Afa-family sequences, the signals of pHvA14 in barley chromosomes tended to appear in the distal regions that do not carry many chromosome band markers. In the telomeric regions these signals always placed in more proximal portions than those of HvRT-family. Based on the distribution patterns of Afa-family sequences in the chromosomes of barley and D genome chromosomes of wheat, we discuss a possible mechanism of amplification of the repetitive sequences during the evolution of Triticeae. In addition, we show here that HvRT-family also could be used to distinguish individual barley chromosomes from the patterns of in situ hybridization.

Biological Evolution↗
PubMed1996

Detection of human T-lymphotropic virus type-I DNA and mRNA in the lymph nodes; using polymerase chain reaction in situ hybridization (PCR/ISH) and reverse transcription (RT-PCR/ISH).

To examine the relationship between human T-lymphotrophic virus type I (HTLV-I) proviral DNA and its expression in the lymph nodes, HTLV-I DNA and tax/rex mRDA were directly amplified by polymerase chain reaction in situ hybridization (PCR/ISH), and reverse transcription (RT)-PCR/ISH [RT-PCR/ISH]. We studied 24 lymph nodes from patients with adult T-cell leukemia/lymphoma (ATLL), incipient ATLL (I-ATLL), and HTLV-I associated lymphadenitis dermatopathic type (HAL-D) and enlarged paracortical type (HAL-EP). In ATLL, 40-60% of the nucleated cells were positive for for HTLV-I proviral DNA by PCR/ISH, while in I-ATLL and HAL, respectively 5-20% and less than 1-5% of cells were positive. The number of mRNA expressing cells was smaller than that of the proviral DNA-positive cells. The mRNA-expressing cells varied in number among the ATLL and I-ATLL cases, while they were only rarely observed in HAL-D and HAL-EP. These results show that HTLV-I infection and activation might increase with malignant transformation of the target T helper cells.

Adult↗
PubMed1996

Epstein-Barr virus infection and associated products (LMP, EBNA2, vIL-10) in nodal non-Hodgkin's lymphoma of human immunodeficiency virus-negative Japanese.

Sixty cases of B-cell nodal non-Hodgkin's malignant lymphoma (B-ML), and 46 cases of T-cell nodal lymphoma (T-ML) were surveyed for Epstein-Barr virus (EBV) genomes, RNA, and associated proteins. We used a Southern blot analysis, polymerase chain reaction (PCR), and EBV-encoded small RNA-1 (EBER-1) in situ hybridization to investigate the presence of EBV. We performed an immunohistochemical study on EBV-related oncoproteins, such as EBV-determined nuclear antigen-2 (EBNA-2), latent membrane protein (LMP), and viral interleukin-10 (vIL-10). In addition, we also analyzed the terminal repetitive sequence of EBV (EBV-TR) to investigate the EBV-infected cell clonality. Non-Hodgkin's lymphomas were grouped into three types by number of EBV-infected cells: I) almost all lymphoma cells showed an EBV presence; II) some scattered lymphoma cells showed an EBV presence; and III) only a few cells showed such a presence, which was probably due to a latent EBV infection. In 25 of 60 B-MLs, EBV-infected cells were found; 7 were type I, 1 was type II, and 17 were type III. In 27 of 46 T-MLs, EBV-infected cells were found; no cases were type I, 5 cases were type II, and 22 cases were type III. Seven B-MLs and 3 T cell lymphomas showed clonal TR bands. Expression of EBNA-2 was found in only three B-MLs, whereas LMP was seen in four B-MLs and six T-MLs. All EBNA-2/LMP-positive cases showed an EBV presence. In B-MLs, expression of EBNA-2 and LMP was detected in almost all lymphoma cells; in T-MLs, however, LMP was found in only a small portion of the lymphoma cells. Expression of IL-10 was closely associated with LMP. In summary, it was thus speculated that EBV infection was associated with the various states of lymphomagenesis.

Antigens, Viral↗
PubMed1996

Classical Hodgkin and Reed-Sternberg cells demonstrate a non-clonal immature B lymphoid lineage: evidence from a single cell assay and in situ hybridization.

Hodgkin and Reed-Sternberg (H & RS) cells are generally considered to be the neoplastic cells of Hodgkin's disease (HD), however such cells are only found in a minority of the lesions. Recently in a few studies on HD, the clonality of H & RS cells was examined, using a single-cell polymerase chain reaction (PCR) examination. To clarify the lineage and clonality of H & RS cells, we performed single cell PCR and in situ hybridization (ISH), and nine cases of classical HD were thus studied. By ISH, the immunoglobulin J chain, and the kappa and lambda light chain were rarely expressed in the H & RS cells, however, no T-cell markers could be detected. The expression of the recombination activating genes (RAG-1, 2) could be determined in the H & RS cells. We isolated CD30+ H & RS cells, CD3 + T cells and CD20 + B cells from suspended materials using a mechanical sorter. We performed single cell PCR in a sorted individual cell, to amplify the complementarity determining region of the Ig heavy chain (IgH) gene and T-cell receptor gamma chain (TCR gamma) gene. In all cases, TCR gamma could be frequently amplified in the T cells, but was only rarely amplified in the H & RS and B cells. In contrast, the IgH was frequently amplified in the H & RS and B cells, but not in the T cells. In addition, the PCR production of the H & RS cells all showed different lengths. The results therefore support the polyclonal nature and immature B lymphoid cell origin of H & RS cells.

B-Lymphocyte Subsets↗
PubMed1996

Philtrum creation in secondary unilateral cleft lip repair.

A new method to create a philtrum in patients with unilateral cleft lip is presented. This modification was primarily used in secondary repair but can also be used in some primary repair cases. The results were satisfactory in most of the patients, at least at 2 years postoperatively.

Cleft Lip↗
PubMed1996

The Sec-1 locus on the short arm of chromosome 1R of rye (Secale cereale).

This paper describes a detailed sequence analysis of the omega-secalin gene array at the Sec-1 locus on the short arm of chromosome 1 of rye. The analysis shows that the genes are separated by 8 kb of spacer sequence and that the gene/spacer units are arranged in a head to tail fashion. The boundaries of the array are identified, and a fragment containing the majority of the genes in the array is separated by PFG analysis. The sequence data of one 9.2 kb gene unit have been determined, and because of the similarity of the gene units within the array these data provide a detailed sequence analysis of 140 kb of the Sec-1 locus. Fluorescence in situ hybridization, using lambda clones isolated for the structural analysis, identifies the position of the array on the rye chromosomes relative to the 5S rRNA genes.

Cloning, Molecular↗
PubMed1995

[Scanning electron microscopic study of the sensory hairs of the organ of Corti using transtympanic infusion of fluids with different osmotic pressure].

Morphological examination of the sensory hairs of the lower half of the 2nd turn of the organ of Corti of the guinea pig after transtympanic infusion of the aseptic fluids with different osmotic pressure was performed using a scanning electron microscope. Group A fluids (0-315 mosmol/l) did not have any injurious effect on the sensory hairs of the OHCs (outer hair cells) or IHCs (inner hair cells). In contrast, Group B fluids (720, 743 mosmol/l) appeared to cause disarrangement of the stereocilia of the OHCs. Alterations were found to be more extensive and severe in the 3rd row of OHCs. Group C fluids (1224, 1248 mosmol/l) induced total loss of sensory hairs in the 3rd and 2nd row of OHC. The remaining stereocilia in the 1st and 2nd rows showed coalescence of adjacent stereocilia. Based on these findings, the authors concluded that there is a close relationship between osmotic induction/deduction by transtympanic infusion of fluids and high osmotic pressure on the hairs of the organ of Corti.

Animals↗
PubMed1994

Lip closing pressure in disabled children: a comparison with normal children.

Lip functions play an important role in the oral stages of feeding. Lip closing is an important early motor act in food acquisition and is essential for controlling chewing and swallowing. To date, there have been few papers on the developmental aspects of lip closing strength when taking in food, especially with regard to disabled children. This investigation was designed to produce an ordinal scale of midline lip pressure measurements for a cross-sectional, age-grouped population of normal children. Developmental changes in lip pressure were then compared with those of two populations of disabled children. Pressure measurements were obtained with a strain gauge transducer that was embedded in a spoon during normal feeding. The study population consisted of 104 normal children ranging in age from 5 months to 5 years, 11 children who showed developmental delay (mean 4.5 years), and 10 children with cerebral palsy (mean 5.0 years). Lip pressure was found to increase steadily from 5 months to 3 years and to increase slightly from 3 to 5 years in the normal population. The developmentally delayed group and the cerebral palsied group produced lip pressures and coefficients of variation below those of the normal 1 to 2-year-old group.

Age Factors↗
PubMed1994

THE RELATIONSHIP BETWEEN THE LENGTH OF THE CUPULAE OF FREE NEUROMASTS AND FEEDING ABILITY IN LARVAE OF THE WILLOW SHINER GNATHOPOGON ELONGATUS CAERULESCENS (TELEOSTEI, CYPRINIDAE)

Free mechanosensory neuromasts of larval fishes have been described as playing a complementary role to vision in feeding behaviour (Disler, 1971; Iwai, 1972a,b). In certain species or under limited conditions, free neuromasts play a major role in detecting prey. The larvae of mottled sculpin Cottus bairdi can feed on Artemia in the dark by using free neuromasts (Jones and Janssen, 1992). Artificially blinded surface-feeding Aplocheilus lineatus can detect insects on the water surface by means of free neuromasts (Muller and Schwarts, 1982; Tittel et al. 1984; Bleckmann, 1988; Bleckmann et al. 1989). Furthermore, vibrations produced by swimming crustaceans are known to be a potent natural stimulus for the lateral line system in the Antarctic fish Pagothenia borchgrevinki (Montgomery and Macdonald, 1987; Montgomery, 1989). We found that larvae of a plankton feeder, the willow shiner Gnathopogon elongatus caerulescens (Sauvage) (Cypriniformes, Cyprinidae), fed on nauplii of Artemia in complete darkness. Ototoxic compounds, such as streptomycin, have been shown to disturb the function of the lateral line organ or free neuromasts (Kaus, 1987; Blaxter and Fuiman, 1989; Janssen, 1990; Jones and Janssen, 1992). Willow shiner larvae treated with streptomycin sulphate no longer feed on Artemia in the dark (Y. Mukai, in preparation). The willow shiner inhabits calm lakes and feeds on zooplanktonic prey (Nakamura, 1949). The larvae show a high sensitivity to minute water displacements. From these observations and from our findings, it appears that larval willow shiner must feed on zooplankton by using free neuromasts in the dark. In larval willow shiner, the vane-like cupulae of the free neuromasts protrude from the body surface and the long cupulae are 100-250 microm in length (Mukai and Kobayashi, 1991). The prey is detected by the free neuromasts as a result of a slight bending of the cupula in response to local water movements. The shape of the cupula, especially its length, must therefore be related to the sensitivity of the free neuromast, as inferred from the results of Coombs and Janssen (1989) and van Netten and Kroese (1989).

Journal Article↗