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Biomedical subjects

Y Machida

Publications and source records attributed to Y Machida.

At least 91 records · Page 5Linked to original sources
PubMed1994

Organ-specific and auxin-inducible expression of two tobacco par A-related genes in transgenic plants.

We have isolated four genomic DNA clones that contain the transcription initiation site of the parA gene(s) from a tobacco genomic library by hybridization with the 5' segment of the parA cDNA previously isolated. They were classified into two types on the basis of their nucleotide sequences. Southern blot analysis indicated that two types of clones were respectively derived from the two parental species of tobacco, Nicotiana tomentosiformis and Nicotiana sylvestris. The genes corresponding to these clones were designated as parAt and parAs, respectively, and the parA cDNA clone was shown to code for mRNA from parAt on the basis of its nucleotide sequence. The 5' regions about 400 nucleotides upstream from the transcription initiation sites of the parAt and parAs genomic clones were highly homologous to one another, but regions further upstream showed no significant similarity. The coding sequence of the GUS (beta-glucuronidase) reporter gene was linked to the 5'-upstream regions of parAt and parAs, and the sites of expression of these fusion genes were examined in transgenic tobacco plants. In the absence of auxins, both fusion genes were expressed in capsules at a late stage of seed development, mature seeds, a root apex and a root-hair zone whereas no significant expression was seen in other organs. Their expression was enhanced by 2,4-dichlorophenoxyacetic acid in most of the organs of tobacco. The results show that expression of these genes is regulated by both organ-specific and auxin-inducible mechanisms.

Amino Acid Sequence↗
PubMed1994

A longitudinal study from prepuberty to puberty of gingivitis. Correlation between the occurrence of Prevotella intermedia and sex hormones.

A longitudinal study of 24 subjects progressing normally from prepuberty to puberty was undertaken to evaluate the effects of sex hormone levels on clinical and microbiologic parameters and on serum antibodies. During elementary school, at the beginning of the longitudinal monitoring, 2 groups, 12 subjects with gingivitis and 12 gingivitis-free subjects, were selected and observed through puberty. Bone ages and self-assessment of secondary sex characteristics were used to confirm puberty. A statistically significant increase in the proportions of Prevotella intermedia including Prevotella nigrescens and serum antibody levels against P. intermedia was seen in gingivitis group throughout the longitudinal study. Serum levels of testosterone in boys and estradiol and progesterone in girls was positively correlated with levels of P. intermedia and P. nigrescens. In puberty, a slight but significant increase in GI scores over prepuberty has been shown; however, there was no significant change in PlI from prepuberty to puberty. Our study confirmed that there was a statistically significant increase in gingival inflammation and in the proportion of P. intermedia and P. nigrescens in puberty relative to the baseline value, except in the gingivitis-free male group. Our findings suggest that these increases are correlated with elevation in systemic levels of the sex hormones.

Adolescent↗
PubMed1994

Infection by Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, and antibody responses at different ages in humans.

This study examined the serum IgG and IgM responses against Porphyromonas gingivalis and 3 serotypes of Actinobacillus actinomycetemcomitans, and the correlations of these responses with age and homologous infection. A total of 90 individuals were included in this study: 40 subjects with gingivitis, 40 periodontally healthy subjects, and 10 adult periodontitis subjects. The subjects in the gingivitis and periodontally healthy groups were divided into 4 stages based on their physiological age: early childhood, school age, puberty, and adult. In the gingivitis group, there was a positive correlation between increase in age and increase in serum IgG antibody levels against P. gingivalis until puberty. However, no statistically significant difference was found between the puberty stage and the adult stage. The average level of IgG antibodies against A. actinomycetemcomitans in the school age gingivitis group was significantly higher than that in the early childhood gingivitis group for all serotypes (p < 0.01). In serotype c, IgG antibody levels in the school age gingivitis group were significantly higher than in the early childhood gingivitis group or the adult gingivitis group (p < 0.01). With both P. gingivalis and A. actinomycetemcomitans, positive correlations between elevated IgG level and infections by these microorganisms were found in the puberty gingivitis and adult periodontitis groups.

Adolescent↗
PubMed1994

Characterization of tobacco protein kinase NPK5, a homolog of Saccharomyces cerevisiae SNF1 that constitutively activates expression of the glucose-repressible SUC2 gene for a secreted invertase of S. cerevisiae.

We have isolated a cDNA (cNPK5) that encodes a protein kinase of 511 amino acids from suspension cultures of tobacco cells. The predicted kinase domain of NPK5 is 65% identical in terms of amino acid sequence to that of the SNF1 serine/threonine protein kinase of Saccharomyces cerevisiae, which plays a central role in catabolite repression in yeast cells. SNF1 positively regulates transcription of various glucose-repressible genes of the yeast, such as the SUC2 gene for a secreted invertase, in response to glucose deprivation: snf1 mutants cannot utilize sucrose as a carbon source. Expression of cNPK5 in yeast cells allowed the snf1 mutant cells to utilize sucrose for growth and caused constitutive expression of the SUC2 gene in wild-type cells even in the presence of glucose, an indication that the NPK5 protein is present in a constitutively active form in S. cerevisiae. On the other hand, expression of cNPK5 failed to suppress the growth defect of the snf4 mutant cells in the presence of sucrose and to induce expression of the SUC2 gene. These results indicate that SNF4 is required for the induction of SUC2 expression by NPK5, as by SNF1, even if NPK5 is constitutively active in S. cerevisiae. The recombinant NPK5 protein is capable of autophosphorylation in vitro in a reaction that requires Mn2+ rather than Mg2+ ions but is inhibited by Ca2+ ions. Both dicotyledonous and monocotyledonous plants have several copies of the NPK5-related gene, which probably constitute a small gene family. NPK5-related genes were found to be expressed in the roots, leaves, and stems of tobacco plants. The high degree of structural conservation and the functional similarity of NPK5 to SNF1 lead us to speculate that NPK5 (or a related kinase) also plays a role in sugar metabolism in higher plants.

Amino Acid Sequence↗
PubMed1993

Genetic evidence for an interaction between the VirA sensor protein and the ChvE sugar-binding protein of Agrobacterium.

Most vir genes of Agrobacterium, which are required for tumorigenicity of the bacterium, are expressed in response to plant phenolics. The induction of vir is markedly enhanced by specific monosaccharides. Signals generated by both types of compound are transduced into Agrobacterium cells via the functions of the VirA membrane-bound sensor protein. A putative sugar-binding protein, known as ChvE, also functions at a step of the enhancement of vir induction by monosaccharides. To investigate the signal pathway of the enhancement by the sugars, we first isolated a virA mutant of Agrobacterium with a base substitution mutation that caused a single amino acid change in the periplasmic domain. The mutant exhibited no enhancement of vir induction by sugar and had severely attenuated tumorigenicity on Kalanchoe leaves. We then isolated two chvE mutants that restored sugar enhancement on the background of this virA mutation. One chvE mutant, which exhibited a higher level of sugar enhancement, restored the tumorigenicity of the virA mutant. Wild-type and suppressor ChvE proteins were localized in the periplasmic space. These results provide genetic evidence for the physical interaction between VirA and ChvE proteins in the periplasmic space of Agrobacterium, which enhances the cytoplasmic signal generated by phenolics. We also discuss the molecular architecture of the operon to which the chvE gene belongs.

Amino Acid Sequence↗
PubMed1993

Molecular characterization of a short interspersed repetitive element from tobacco that exhibits sequence homology to specific tRNAs.

We have characterized a family of tRNA-derived short interspersed repetitive elements (SINEs) in the tobacco genome. Members of this family of SINEs, designated TS, have a composite structure and include a region structurally similar to a rabbit tRNA(Lys), a tRNA-unrelated region, and a TTG repeat of variable length at the 3' end. Southern blot hybridization, together with a search of the GenBank data base, showed that various plants belonging to the families Solanaceae and Convolvulaceae contain sequences homologous to the TS family in the introns and flanking regions of many genes, whereas Arabidopsis in the family Cruciferae and several species of monocoytledonous plants do not. The TS family is widely involved in structural and genetic variations in the genomes of many plants that belong to the order Tubiflorae. All of nine sequences identified in a data base search are truncated at their 5' regions and lack the tRNA-related region of the TS family. We characterized the entire sequence of the members of the TS family and found that this family can be categorized as a member of a group of SINEs with a tRNA(Lys)-like structure, as can several animal SINEs. The TS family can be divided into two major subfamilies by analysis of diagnostic positions, and one of the subfamilies is clearly younger than the other. Amplification of many copies of the full sequence of the younger subfamily occurred during the recent evolution of the tobacco lineage. We also discuss mechanisms that could be involved in the generation of SINEs in animals and also in plants.

Base Sequence↗
PubMed1993

NPK1, a tobacco gene that encodes a protein with a domain homologous to yeast BCK1, STE11, and Byr2 protein kinases.

We have isolated a cDNA (cNPK1) that encodes a predicted protein kinase of 690 amino acids from suspension cultures of tobacco cells. The deduced sequence is closely related to those of the protein kinases encoded by the STE11 and BCK1 genes of Saccharomyces cerevisiae and the byr2 gene of Schizosaccharomyces pombe. STE11 and Byr2 function in the yeast mating pheromone response pathways, and BCK1 acts downstream of the yeast protein kinase C homolog encoded by the PKC1 gene, which is essential for normal growth and division of yeast cells. Overexpression in yeast cells of a truncated form of cNPK1, which encodes only the putative catalytic domain, replaced the growth control functions of BCK1 and PKC1 but not the mating pheromone response function of STE11. Thus, the catalytic domain of NPK1 specifically activates the signal transduction pathway mediated by BCK1 in yeast. In tobacco cells in suspension culture, the NPK1 gene is transcribed during logarithmic phase and early stationary phase but not during late stationary phase. In a tobacco plant, it is also transcribed in stems and roots but not in mature leaves, which rarely contain growing cells. The present results suggest that a signal transduction pathway mediated by this BCK1- and STE11-related protein kinase is also conserved in plants and that a function of NPK1 is controlled at least in part at a transcriptional level.

Amino Acid Sequence↗
PubMed1993

Effect of pretreatment of skin with cyclic monoterpenes on permeation of diclofenac in hairless rat.

The promoting effect of d-limonene and l-menthol on the percutaneous absorption of nonionized and ionized diclofenac (DF) was investigated employing a pretreatment method. After the pretreatment of hairless rat skin with an ethanol buffer solution containing terpenes, the permeation study was performed in vitro. The permeability coefficients of nonionized (Pn) and ionized diclofenac (Pi) were calculated under the assumption that the total flux was composed of individual fluxes of nonionized and ionized DF. In the case of pretreatment with d-limonene, both Pn and Pi values were increased dramatically. However, the promoting magnitude was not affected by an extension of the pretreatment period. In contrast, when the skin was pretreated with l-menthol, the Pn and Pi values increased gradually as the pretreatment period increased. Based on the measurement of the solubility of terpenes in the pretreatment solution, the difference in promoting efficiency could arise from the difference in thermodynamic activity of the terpenes.

Animals↗
PubMed1993

Evaluation of bioavailability upon oral administration of phytonadione preparations in beagle dogs.

Bioavailability of phytonadione was investigated after oral administration to beagle dogs. The administrations of phytonadione in a surfactant solution (preparation a) and in an oleic acid solution (preparations b) greatly increased the bioavailability of phytonadione. The AUCs of preparations a and b were about 2.5 times larger than those of commercially available tablets A and C. This result well corresponded to the results of the dissolution test previously reported. The absorption of phytonadione from the gastro-intestinal tract was affected by food and the bioavailability was largely increased under non-fasted conditions. However, a large scatter was observed in the data, and it was found that the evaluation of the bioavailability of phytonadione preparations could not be done in non-fasted animals.

Administration, Oral↗
PubMed1993

Investigation on rectal absorption of indomethacin from sustained-release hydrogel suppositories prepared with water-soluble dietary fibers, xanthan gum and locust bean gum.

Sustained-release hydrogel suppositories prepared with water-soluble dietary fibers, xanthan gum and locust bean gum, were evaluated as a vehicle for rectal administration of indomethacin (IMC) in rabbits. The drug plasma levels were compared with those after rectal administration of commercial suppositories. When the commercial suppositories were given to rabbits, the plasma concentration reached the maximum level at 30 min after administration followed by a quick reduction, while no sharp peak of plasma levels was seen with the hydrogel suppositories. In particular, the plasma levels observed with the hydrogel suppositories of 1% (w/v) gum concentration were sustained much longer than those after dosing with the commercial suppositories; the mean residence times had higher values without a decrease in the area under the plasma concentration vs. time curves. Histopathological study showed good biological safety of the hydrogel suppositories to the rectal mucosa. These results suggested that the IMC hydrogel suppositories prepared with xanthan gum and locust bean gum were a practical rectal preparation with prolonged action and reduced side effects.

Animals↗
PubMed1993

Site-dependent effect of aprotinin, sodium caprate, Na2EDTA and sodium glycocholate on intestinal absorption of insulin.

In order to determine an advantageous site for intestinal insulin absorption, the hypoglycemic effects of insulin after administration to the duodenum, the jejunum, the ileum and the colon were investigated using an in situ loop method. Insulin solution was administered to the various loops of fasted rats with or without aprotinin (AP) as a protease inhibitor, or absorption enhancers such as sodium caprate, Na2EDTA or sodium glycocholate. An obvious hypoglycemic effect of insulin alone was seen only in the ileum loop washed with phosphate buffered saline. When coadministered with AP, the most remarkably amplified effect was again observed in the ileum. In the ileum, the area under the serum insulin levels vs. time curve from 0 to 4 h was linearly related to the logarithm of the AP dose. Both sodium caprate and Na2EDTA significantly promoted the hypoglycemic effect of insulin at all sites, and their intensity increased towards the distal regions of the intestine. On the other hand, sodium glycocholate improved only colonic insulin efficacy. These results suggest that the ileum seems to be the most useful region in the small intestine for insulin absorption; however, insulin must be protected from proteolysis to enhance its absorption. In addition, the insulin efficacy could be increased by absorption promoters more effectively in the colon than in the small intestine.

Animals↗
PubMed1993

[Development of topical drug delivery systems utilizing polymeric materials].

Topical drug delivery is important from the view points of improvement of therapeutic effect and reduction of systemic side effects. Utilization of polymeric materials seemed to be as a key for the development of new topical dosage forms including targeting drug delivery systems. Adriamycin ointment for local chemotherapy to breast cancer prepared using polyethylene glycol, ammonium polyacrylate and hydroxypropyl cellulose (HPC) according to an optimum formulation showed an excellent clinical effect in spite of a decreased drug content. Double-layered mucoadhesive sticks for the treatment of uterine cervix cancer were prepared by direct compression of powder mixture of bleomycin, HPC and carboxyvinyl polymer (CP). Drug release property of the sticks could be controlled by the weight of outer layer, drug combining ratio to each layer and coating of core layer. The results suggested a possibility of a "once-a-week" treatment that is preferable for the patients. Magnetic granules for the treatment of esophageal cancer were prepared using ferrite, HPC and CP. Magnetic guidance and retainment of the granules on esophageal mucosa were confirmed using rabbits in vivo. Buoyant sustained release preparations were prepared using chitosan, soybean protein, HPC and other polymers. Usefulness of the buoyant preparations was suggested from the results in vitro and in vivo. Insulin microspheres (IMS) for targeting delivery to the small intestine were prepared by the newly developed method. Employment of enteric coating material (Eudragit) and combination of protease inhibitor protected insulin from enzymatic attack and gave decreased levels of blood glucose by oral administration.

Administration, Topical↗
PubMed1993

Evaluation of the conjugate between N4-(4-carboxybutyryl)-1-beta-D-arabinofuranosylcytosine and chitosan as a macromolecular prodrug of 1-beta-D-arabinofuranosylcytosine.

The conjugate of N4-(4-carboxybutyryl)-1-beta-D-arabinofuranosylcytosine (glu-ara-C) with chitosan, named chi-glu-ara-C, was prepared and evaluated on its utility as a macromolecular prodrug. 1-beta-D-Arabinofuranosylcytosine (ara-C) was predominantly regenerated gradually from chi-glu-ara-C in 1/15 M phosphate buffer, pH 7.4, at 37 degrees C, and its amount after 7 d incubation was 56% (w/w). The antitumor effect was evaluated by an increase in the lifespan (ILS) of the mice bearing P388 leukemia at the single administration at 24 h after inoculation. Ara-C exhibited an ILS of 3.4% at a dose of 100 mg/kg, and the survival time was not significantly different from that of the control. However, chi-glu-ara-C showed an ILS of 60.7% at a dose of 88 mg eq ara-C/kg, and the survival time was significantly different from that of the control. Chitosan and chi-glu-ara-C were observed to cause the side effects judged from the body weight loss to a certain extent. However, chi-glu-ara-C was recognized to be useful because of its effectiveness.

Animals↗
PubMed1993

[Severely mentally and physically disabled in Gunma Prefecture].

We studied severity, causes of the disabilities, complications due to disabilities, and past histories of the severely mentally and physically disabled whose guardians or parents were residents in Gunma prefecture on September 1, 1991. This survey was made on severely disabled schoolboys and girls aged 6 to 15 years with IQ of 35 or less who could not stand or walk, even if they were supported. The number of them was 133 at the moment of this survey. Asphyxia at birth and congenital but unknown etiology were the major causes in the institutionalized cases, but cerebral palsy of unknown etiology and "no information" were frequent in the cases cared at home. The institutionalized patients showed severer mental retardations, palsies and disabilities on activities of daily lives, though not with statistically significant difference, and contained significantly larger number of cases fed by gastric tube or having meals as blended foods and significantly smaller number of cases having conventional meals than cases cared at home. Furthermore, epilepsies or constipations as complications were found more frequently in institutionalized cases than in cases cared at home with significant difference. On past history, relatively large numbers of the disabled suffered from various infectious diseases, especially from measles, varicella, mumps and pneumonia. Histories of measles, varicella and mumps showed higher frequency in institutionalized cases than in cases cared at home, though not with significant difference. On the cases cared at home, some of them did not have their home doctors, though in small numbers.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

[Prevalence rate of the severely mentally and physically disabled children in Gunma Prefecture].

Children were studied on distribution of the severely mentally and physically disabled aged 6 to 15 years in Gunma Prefecture as of September 1, 1991. They were all bedridden or able to sit, but could not stand or walk, even with support. Today, all the Japanese should attend elementary school as well as junior high school. Even though they are severely disabled, they are all registered by the regional Board of Education, where we could get exact data on the disabled children throughout Gunma Prefecture. We collaborated with the regional Board of Education, the Child Consultation Center of Gunma Prefecture and 6 hospitals for the disabled. A total of 129 cases were found. The population of schoolboys and schoolgirls aged 6 to 15 years of age was 236,000, most of whose guardians were residents of this prefecture, and the prevalence rate was 0.55/1000. Among 129 cases, 53 cases (41.1%) were institutionalized and 76 cases (58.9%) were cared for at home. Fifty-nine cases (45.7%) were males and 70 cases (54.3%) were females. Thirty-four cases were aged 6 to 7 years (26.4%) and older cases declined probably due to premature deaths. Ninety-four of the 129 cases (72.9%) were bedridden and mentally retarded with IQs 20 or less.

Adolescent↗
PubMed1992

Prospective study on maternal, intrauterine, and perinatal infections with cytomegalovirus in Japan during 1976-1990.

Since 1976, sera obtained serially from 10,218 pregnant women during the first, second, and third trimesters of gestation and cord sera were tested for CMV complement-fixing (CF) and immunofluorescent (IF) antibodies. CMV IgG-IF antibody was positive in 9,735/10,218 (95%) in the first trimester, and a significant rise of CF antibodies during pregnancy was found in 70/9,206 (0.76%) of the seropositive group and in 5/438 (1.14%) of the seronegative group. IgM antibody was found in 6/9,206 (0.06%) of seropositive women during the first trimester and in 7/70 (10.0%) of seropositive mothers with CF antibody rise and in 4/5 of seroconverted mothers of the seronegative group, suggesting that the incidence of primary infection with CMV during pregnancy was approximately 1% of susceptible women. All the mothers with immune response had infants with neither viruria nor IgM antibody in the cord blood, whereas seropositive mothers without an immune response had infants with viruria (7/1,826; 0.4%) or with IgM antibody in the cord blood (6/9,136; 0.06%). None of these 13 babies, shedding CMV or with IgM IF antibody, had physical or mental retardation. CMV IgG-IF antibody was present in almost 80% of infants between 7 and 12 months of age in 1988, suggesting that perinatal or postnatal CMV infection may occur in infants born to seropositive mothers in 70-80% of pregnancies.

Antibodies, Viral↗
PubMed1992

Factors affecting prednisolone release from hydrogels prepared with water-soluble dietary fibers, xanthan and locust bean gums.

The release behavior of prednisolone from hydrogels prepared with xanthan and locust bean gums was investigated. Newly developed equipment was employed in order to increase the gum concentration in the hydrogels. The apparent release rate of prednisolone from the hydrogels decreased with increasing gum concentration, suggesting that the diffusion of drug molecules was mainly controlled by the density of the three-dimensional network structure in the matrix. The effect of additives such as glycerin and sucrose on the release behavior of prednisolone was also investigated in detail. Drug release was significantly lowered by the addition of these compounds to these hydrogels. A linear relation was observed between the apparent release rate of prednisolone and microscopic viscosity of the hydrogels. These results indicated that the drug release could be controlled not only by the density of the network structure but also by the microscopic viscosity of the hydrogels.

Delayed-Action Preparations↗