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Biomedical subjects

N Takamatsu

Publications and source records attributed to N Takamatsu.

At least 37 records · Page 2Linked to original sources

Effect of initial periodontal therapy on the frequency of detecting Bacteroides forsythus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans.

BACKGROUND: Porphyromonas gingivalis, Bacteroides forsythus, and Actinobacillus actinomycetemcomitans have been described as periodontopathic bacteria, and their presence in subgingival pockets can lead to development of periodontal disease. Until now, clinical parameters have been used to evaluate the effect of conventional periodontal treatment without microbiological parameters. The present study examined the microbiological effects of initial periodontal therapy using DNA probes and the polymerase chain reaction (PCR). METHODS: Twenty-six patients with periodontitis, 10 males and 16 females, were given instructions regarding oral hygiene, then thoroughly treated by conventional scaling and root planing. Bacterial samples were collected on paper points from 4 sites per patient at baseline and after initial therapy (total: 104 sites). Clinical parameters including probing depth, attachment level, and bleeding on probing were also recorded for each site at baseline and after therapy. A DNA probe kit was used to monitor the frequency of B. forsythus, P. gingivalis, and A. actinomycetemcomitans, the last of which was identified by PCR. RESULTS: At baseline, B. forsythus was the bacterium most frequently detected. DNA probe analysis also showed that more than half of the sites were colonized by both B. forsythus and P. gingivalis. Initial therapy resulted in significant clinical improvement such as significant reduction in the frequency of B. forsythus and P. gingivalis detected using the DNA probe. A. actinomycetemcomitans was difficult to detect using the DNA probe, but PCR indicated that levels of A. actinomycetemcomitans did not significantly decrease. CONCLUSIONS: These results indicate that initial conventional therapy can eliminate B. forsythus and P. gingivalis, but not A. actinomycetemcomitans. When levels of these bacteria decreased to below-detectable levels, clinical improvement was significant. These results indicate that monitoring levels of these three periodontopathic bacteria may render periodontal therapy more effective and accurate.

Adolescent↗

Rainbow trout Sox24, a novel member of the Sox family, is a transcriptional regulator during oogenesis.

We isolated a cDNA clone encoding a novel SRY-type HMG box (Sox) protein, designated Sox24, from a rainbow trout ovary cDNA library. On the basis of the HMG box amino acid sequence, Sox24 can be categorized into the same subgroup of Sox proteins as SOX4, SOX11, and SOX22. The proteins in this group also share a highly conserved sequence at the C-terminus. The Sox24 mRNA is expressed at high levels in the ovary, and in-situ hybridization localized its expression to oocytes. The recombinant protein containing the Sox24 HMG box region bound to an AACAAT sequence strongly in a gel retardation assay. Upon co-transfection into CHO cells, the full-length Sox24 transactivated transcription from a reporter plasmid through the AACAAT binding motif. We used GAL4/Sox24 chimeras with the DNA binding domain of yeast GAL4 at the N-terminus to map the transactivation function to the C-terminal region, which included the conserved sequence. These results suggest that Sox24 plays a role as a transcriptional regulator during oogenesis.

Amino Acid Sequence↗

cDNA cloning of a novel rainbow trout SRY-type HMG box protein, rtSox23, and its functional analysis.

We have isolated a cDNA clone for a new member of Sox genes, termed rtSox23, from a rainbow trout ovary cDNA library. rtSox23 mRNA was notably expressed in ovary and brain. rtSox23 contains a leucine zipper in addition to an SRY-type HMG box. Although the recombinant HMG box region protein of rtSox23 could bind to an AACAAT sequence, the full-length rtSox23 could form a homodimer and did not bind to the sequence. Furthermore, using a two-hybrid system, we have isolated a cDNA clone encoding a protein that bound to the leucine zipper region of rtSox23. This protein was the rainbow trout homologue of mouse nucleoporin p62, which is a component of the nuclear pore complex in nuclear envelope. The rainbow trout p62 mRNA was also prominent in ovary and brain. Taken together, these results suggest that the rainbow trout p62 associates with rtSox23 in vivo and modulates the function of rtSox23.

Amino Acid Sequence↗

cDNA cloning of a new member of the FTZ-F1 subfamily from a rainbow trout.

We describe here cDNA cloning of an orphan nuclear receptor family member, tFZR1, which has a FTZ-F1 box. The amino acid sequences of the zinc finger domain and the FTZ-F1 box has 92.8% and 100% identity, respectively, with those of zebrafish FTZ-F1. On the other hand, the overall homology between tFZR1 and zebrafish FTZ-F1 is low (33.0%). The results indicate that tFZR1 is a new member of fushitarazu factor 1 (FTZ-F1) subfamily.

Amino Acid Sequence↗

Subgingival distribution of periodontopathic bacteria in adult periodontitis and their susceptibility to minocycline-HCl.

The purpose of this study was to investigate the distribution of several periodontopathic bacteria in adult periodontitis, their in vitro susceptibility to minocycline-HCl, and whether the efficacy of the drug changes with a decrease in bacterial susceptibility. Twenty-one patients (43 to 75 years old) with 62 periodontal lesions from pockets > or =4 mm participated in the study. After subgingival sampling, an ointment containing 2% minocycline-HCl was applied locally to the selected pockets once a week for 4 weeks. The lesions were clinically examined after 1 and 4 weeks of administration. The distribution of the subgingival microorganisms included Capnocytophaga sputigena (37.1%), Prevotella intermedia (22.6%), Porphyromonas gingivalis (22.6%), Fusobacterium nucleatum (20.1%), Actinobacillus actinomycetemcomitans (9.7%), and Eikenella corrodens (4.8%). The distribution was complex, with 76.8% of the sites containing 1 to 3 bacterial spieces. The minimum inhibitory concentration (MIC) of minocycline-HCl for each organism showed that most were inhibited by a minocycline-HCl concentration equal to or less than the MIC for reference strains. However, some clinical strains of Prevotella intermedia seemed to exihibit low susceptibility to minocycline-HCl. There were no significant differences among sites with strains exhibiting low or normal susceptibility to minocycline-HCl. The concentration of the drug applied to deep periodontal pockets inhibited the growth of most of the microorganisms investigated in this study.

Administration, Topical↗

[The new colorimetric assay (WST-1) for cellular growth with normal aging and Alzheimer's disease].

A new tetrazolium compound, WST-1, a sodium salt of 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate, was introduced into clinical chemistry. The compound's dehydrogenase activities are useful in the colorimetric determination of the bioreducibility of cells. It proved to have a sensitivity similar to MTT, and produces a highly colored formazan dye under reductive conditions. Compared with the MTT assay, we found the WST-1 assay to be more sensitive, more convenient, and more exact. We used the new assay to study aging-related changes in human lymphocyte blastformation. Four groups of healthy adults aged 40-49 years (N = 40), 50-59 years (N = 40), 60-69 years (N = 40), 70-76 years (N = 40) were examined. Moreover, we studied lymphocytes from 16 patients (47-74 years) with probable Alzheimer's disease. For healthy controls, a negative correlation (r = -0.3108, p < 0.05) between age and enzyme activity was noted. The regression equation was: Y = -0.0085X + 1.473 where X and Y designate the age of the individual and enzyme activity (absorbance), respectively. On the other hand, for patients with Alzheimer's disease, age and enzyme activity did not correlate. These results indicate that the WST-1 assay may be useful in the study of changes associated with aging, and also more so than the MTT assay.

Adult↗

Expression of multiple alpha1-antitrypsin-like genes in hibernating species of the squirrel family.

In the chipmunk, a mammalian hibernator, a 140 kDa protein complex found in the blood, drastically decreases in concentration during hibernation. This complex contains four species of proteins, HP-20, -25, -27 and -55. In the present study, cDNA clones coding for the chipmunk HP-55 were isolated from a liver cDNA library. Sequence analysis revealed that HP-55 is produced as a precursor protein of 413 amino acids (aa), that it has a signal peptide of 24 aa, and that it contains four potential N-glycosylation sites. The deduced aa sequence shows 63% identity with that of rat alpha1-antitrypsin (alpha1-AT); however, the sequence corresponding to the reactive center P1-P1' residues was found to be Met-Leu, whereas it is Met-Ser in the rat alpha1-AT. During screening of the chipmunk liver cDNA library, four other related classes of cDNA clones were obtained, each also coding for an alpha1-AT-like protein. In spite of more than 86% overall aa sequence identity among the five chipmunk alpha1-AT-like proteins, they are highly divergent in the putative reactive center region; the putative P1-P1' sequences are Met-Leu (HP-55 or CM55-ML), Met-Met (CM55-MM), Met-Ser (CM55-MS), Ser-Ile (CM55-SI) and Ser-Thr (CM55-ST). Each of the alpha1-AT-like protein mRNAs was expressed in chipmunk liver, and the HP-55 mRNA level was greatly reduced during hibernation. Genomic Southern blot analysis and screening of a liver cDNA library from another hibernating squirrel species, the ground squirrel, also revealed expression of multiple members of the alpha1-AT gene family, whereas analysis of a cDNA library from a non-hibernating species, the tree squirrel, found only a single alpha1-AT gene.

Amino Acid Sequence↗

Rainbow trout SOX9: cDNA cloning, gene structure and expression.

We have isolated and characterized rainbow trout SOX9 cDNA and genomic clones. The cDNA encodes a protein of 488 amino acids (aa) with an HMG box and has 70% aa sequence identity with human SOX9. The rainbow trout and human SOX9 genes show a similar gene structure, and the two introns in the coding region are located at conserved positions. In rainbow trout, the SOX9 mRNA was prominent in testis and brain.

Amino Acid Sequence↗

The Xenopus Sox3 gene expressed in oocytes of early stages.

We have isolated an SRY-type HMG box (Sox) cDNA, XSox3, from a Xenopus immature ovary cDNA library. The XSox3 cDNA contains an open reading frame (ORF) of 309 amino-acid residues, showing 62.7 and 77.0% homology with human and chicken Sox3 proteins, respectively. We also showed that the XSox3 gene is composed of a single exon by sequence analysis of the genomic clone and the determination of the transcription start site of the XSox3. The XSox3 mRNA was detected only in ovary and was at a higher level in immature ovary than in mature ovary. During oocyte maturation, the XSox3 mRNA was most abundant in stage I oocytes, and the XSox3 protein was detected in stage I and II oocytes. Recombinant XSox3 protein produced in Escherichia coli bound specifically to sequences containing the binding motif for the HMG box of SRY or SOX proteins, AACAAT or AACAAAG, demonstrating its sequence-specific DNA binding property. Taken together, these results indicate that the XSox3 protein may participate in early oogenesis of Xenopus as a transcription factor.

Amino Acid Sequence↗

Human intestinal permeability of piroxicam, propranolol, phenylalanine, and PEG 400 determined by jejunal perfusion.

PURPOSE: To determine the human jejunal permeabilities of compounds utilizing different transport mechanisms using a regional perfusion approach and to establish a standard procedure for determining drug permeability class to be used for the establishment of drug product bioequivalence standards. METHODS: Six healthy male volunteers participated in this study. A multi-lumen perfusion tube was inserted orally and positioned in the proximal region of the jejunum. A solution containing piroxicam, phenylalanine, propranolol, PEG 400 and PEG 4000 was perfused through the intestinal segment at a rate of 3.0 ml/min. Perfusate samples were quantitatively collected every 10 minutes for two 100 minute periods with an intermediate wash out period to determine intra and intersubject variation. RESULTS: The mean P(eff) (+/-SD) of piroxicam, phenylalanine, propranolol, and PEG 400 were 10.40 +/- 5.93, 6.67 +/- 3.42, 3.59 +/- 1.60, 0.80 0.46 x 10(-4) cm/sec, respectively. The coefficient of variation for the intersubject variability, first and second perfusion periods were: piroxicam, 60.5% and 57.1%; phenylalanine, 52.8% and 57.8%; propranolol, 62.1% and 44.6%; and PEG 400, 81.7% and 42.3%, indicating a slightly lower CV for the second perfusion period in the same subject. The intrasubject CV's between the two perfusion periods were: 19.4%, 21.3%, 23.6% and 41.0% respectively, indicating a smaller intraindividual variation for all compounds studied. CONCLUSIONS: Piroxicam, a nonpolar drug exhibited the highest permeability of the compounds studied. The intrasubject CV was lower than the intersubject CV, indicating consistent permeability estimation within subjects. The methodology is useful for permeability estimation regardless of absorption mechanism and can be used to establish a consistent data base of human permeabilities for estimation of human drug absorption and for establishing the biopharmaceutic permeability class of drugs.

Adult↗

Three cases of pyogenic sacro-iliitis, and factors in the relapse of the disease.

Pyogenic sacro-iliitis (PS) is a rare disease in childhood. Three cases of PS are reported that were difficult to diagnose. Scintigraphy and magnetic resonance imaging (MRI) were useful for diagnosis. One patient suffered from an episode of relapse. Seventeen other cases of PS were reviewed in the literature to investigate the incidence of abnormal imaging findings and various factors in disease relapse. It was found that the incidence of abnormal findings by scintigraphy was significantly higher than that by computed tomography (P = 0.0057). The duration of intravenous antibiotic administration of the relapse group (14.7 +/- 4.7 days) was significantly shorter than that of the non-relapse group (24.3 +/- 10.7 days; P = 0.0376). The statistical analysis suggested that intravenous antibiotic administration is necessary at least for 20 days to prevent a relapse of PS.

Adolescent↗

[The effect of periodontal initial therapy on detection frequency of Bacteroides forsythus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans--detection by DNA probe and PCR methods].

The purpose of this study was to examine microbiological changes after periodontal initial therapy by DNA probe and polymerase chain reaction (PCR) assays in addition to clinical evaluation. Twenty-six periodontitis patients, 10 males and 16 females, were selected. All subjects were treated by scaling and root planing following oral hygiene instruction. Bacterial samples were collected with paper points from a total of 104 sites, 4 sites in each patient, at baseline and after initial therapy. DNA probe analysis was used to monitor Bacteroides forsythus and Porphyromonas gingivalis. PCR analysis was performed to identify the presence of Actinobacillus actinomycetemcomitans. The initial therapy resulted in significant clinical improvements as assessed by clinical parameters including pocket depth, attachment level and bleeding on probing. After the therapy, the detection frequency of B. forsythus and P. gingivalis was significantly reduced, but the detection frequency of A. actinomycetemcomitans did not significantly decreased. These results indicated that initial therapy can bring about elimination of B. forsythus and P. gingivalis by means of conventional techniques, but not of A. actinomycetemcomitans. When complete elimination of these microorganisms was obtained, significant clinical improvement was found. Monitoring the changes of three bacteria may provide a more effective modality.

Adolescent↗

Abnormal excitability of the corticospinal pathway in patients with amyotrophic lateral sclerosis: a single motor unit study using transcranial magnetic stimulation.

The pathophysiology of corticospinal tract degeneration in amyotrophic lateral sclerosis (ALS) was investigated by studying the effect of transcranial magnetic stimulation on discharge characteristics of single motor units during voluntary activation. The motor units were recorded from the first dorsal interosseus muscles of 12 patients with ALS, 14 healthy subjects, 12 patients with upper motor neuron lesions and 9 with pure lower motor neuron diseases. More than 100 magnetic stimuli were delivered over the scalp during minimal muscle contraction. The occurrence of motor unit discharges was plotted in a peristimulus time histogram. An increase in discharge probability at latencies of 20-30 msec, that represents monosynaptic activation (primary peak) was found in normal units. Motor units from ALS patients with short disease durations had significantly increased discharge probabilities in the primary peak (P < 0.001). Motor units from 4 ALS patients with upper motor neuron signs showed double primary peaks: an initial synchronized peak followed by a dispersed peak. The latter was ascribed to a slow corticospinal pathway, which remains undetected or is functionally insignificant in healthy subjects. We conclude that the excitabilities of the surviving corticospinal tract pathways are abnormally increased in ALS, especially in the early stage.

Adolescent↗

[Crystal forms, improvements of dissolution and absorption of poorly water-soluble (R)-1-[2,3-dihydro-1-(2'-methylphenacyl)-2-oxo-5-phenyl-1H-1, 4-benzodiazepin-3-yl]-3-(3-methylphenyl)urea (YM022)].

Polymorphs of (R)-1-[2,3-dihydro-1-(2'-methylphenacyl)-2-oxo-5-phenyl-1H-1,4- benzodiazepin-3-yl]-3-(3-methylphenyl)urea (YM022) were investigated. Two crystalline forms (alpha- and beta-forms) of YM022 were confirmed by powder X-ray diffractometry and differential scanning calorimetry. alpha- And beta-forms were obtained by recrystallization from ethanol and ethanol: water (5:1), respectively. Amorphous YM022 was obtained by spray drying of YM022 methanol solution. Since both crystalline and amorphous YM022 were sparingly soluble in water, solubilization of YM022 by solid dispersion and wet grinding methods were performed. In vitro dissolution study and in vivo absorption study in dogs were carried out using spray-dried solid dispersion, heat-treated solid dispersion and mechanical mixture. Spray-dried solid dispersion and heat-treated solid dispersion showed enhanced bioavailability, whereas mechanical mixture showed no improvement.

Animals↗

Effects of aging on P3 components of somatosensory event-related potentials elicited by different stimulus conditions. Conventional averages and single-trial analysis.

Effects of aging on P3 components of somatosensory event-related potentials (ERPs) under different stimulus conditions were studied in healthy subjects using conventional averages and single-trial waveform analysis. The P3 component elicited by target stimuli (target P3) and that elicited by rare nontarget stimuli (nontarget P3) showed significant latency prolongation with age at the rate of 1.6 and 2.0 ms/year, respectively. The nontarget P3 showed a greater trial-to-trial fluctuation of single-trial latency than the target P3, probably because 'passive attention' was more liable to fluctuate. The variabilities of single-trial target P3 latency and of single-trial nontarget P3 latency were not correlated with age. Thus, the P3 latency prolongation with age might not be attributable to fluctuation in latency resulting from impairment of attention with aging.

Adult↗

[Evaluation of non radioactive DNA probe (Affirm DP) for detecting periodontopathic bacteria].

The purpose of this investigation was to evaluate the potential of application of non-radioactive DNA probe method (Affirm DP) in clinical diagnosis and evaluation of treatment efficacy. Sensitivity and specificity of two different cut-off levels (10(4), 5 x 10(5)) of the method with bacterial culture in detection of Bacteroides forsythus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans were compared. Those bacteria were obtained from bacterial cultures and clinical samples from healthy subjects and patients with periodontal disease. The results showed that detection percentage of DNA probe method with 10(4) cut-off was equal to or higher than the culture method for Bacteroides forsythus and Porphyromonas gingivalis detection. Therefore, it is suggested to be a useful chairside DNA probe kit as an aid in clinical diagnosis and evaluation of treatment efficacy.

Aggregatibacter actinomycetemcomitans↗

Molecular cloning of the defense factor in the albumen gland of the sea hare Aplysia kurodai.

Aplysianin-A, an antibacterial glycoprotein in the albumen gland of the sea hare Aplysia kurodai, inhibited the growth of both Gram-positive and Gram-negative bacteria. Aplysianin-A cDNA clones were isolated from an albumen gland cDNA library. Sequence analysis reveals that aplysianin-A is produced as a precursor protein of 556 amino acid residues with a signal peptide of 19 amino acid residues and contains 6 potential N-glycosylation sites. Aplysianin-A mRNA was expressed tissue-specifically in the albumen gland. Homology search reveals that aplysianin-A has a 50% overall amino acid sequence homology to achacin, an antibacterial glycoprotein of the giant African snail Achatina fulica.

Amino Acid Sequence↗

A rainbow trout SRY-type gene expressed in pituitary glands.

A Sox (SRY-type HMG box) gene, designated SoxP1, was isolated from a cDNA library made from pituitaries of immature rainbow trout. Sequence analysis indicated that the cDNA had an open reading frame encoding 467 amino acid residues containing a DNA binding motif, known as the high mobility group (HMG) box. Northern blot analysis showed trout SoxP1 mRNA was detected in pituitaries and gonadal tissues, but not in liver, spleen, and heart. In pituitaries, trout SoxP1 mRNA was more abundant in immature fish than in mature fish. Gel shift retardation analysis indicated that the recombinant HMG box protein of SoxP1 produced in E. coli had a DNA binding property for an AACAAT or AACAAAG sequence. These findings suggest that the trout SoxP1 protein may play certain roles in growth or maturation in pituitaries as a transcription factor.

Amino Acid Sequence↗