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Biomedical subjects

N Ozawa

Publications and source records attributed to N Ozawa.

At least 37 records · Page 2Linked to original sources
PubMed1996

Evaluation of human skin irritation by carboxylic acids, alcohols, esters and aldehydes, with nitrocellulose-replica method and closed patch testing.

Closed patch testing and the nitrocellulose-replica method are performed as useful clinical methods for the evaluation of human skin irritation by cosmetics and topical medicaments. Comparison of the sensitivity between microscopic scoring by nitrocellulose-replica method and visual scoring by closed patch test in the detection of skin irritation, however, has not been well studied with statistical analysis. Here, we evaluated human skin irritation by carboxylic acids, alcohols, esters and aldehydes, with different chain length (C8-C18), using both methods. The results of closed patch testing showed that, although the score of skin irritation for carboxylic acids (C8, C12), alcohols (C8) and aldehydes (C8), tested at a concentration of 0.5 m-2.0 m, significantly increased with increasing concentration of the test compounds, ester compounds scarcely caused any irritation on the surface of the skin occluded. In addition, an increase of carbon chain length in the test compounds made it impossible to detect skin irritation. In contrast, the nitrocellulose-replica method could evaluate skin reactions against very weak irritants that gave no macroscopic alterations on the skin surface in the closed patch test. However, the scoring system is somewhat subjective and should be improved to make the analysis more objective.

Adult↗
PubMed1996

Effects of gonadotropin-releasing hormone agonist (GnRHa) and ovariectomy on leukocyte subpopulations in rats with autotransplanted endometrium.

PROBLEM: To determine the effect of estrogen deficiency on populations of immune cells in rats with autotransplanted endometrium. METHOD OF STUDY: Autologous endometrial segments were implanted in the peritoneum of rats. Leukocyte subsets in the peripheral blood (PB) and peritoneal fluid (PF) of autotransplanted rats treated with gonadotropin-releasing hormone agonist (GnRHa) or ovariectomy were analyzed by flow cytometry. RESULTS: Endometrial growth was suppressed in association with increases in natural killer (NK) cells and macrophages in rats treated surgically or medically after endometrial implantation compared with autotransplanted-only rats. Normal rats treated with GnRHa showed no significant alterations in PB and PF leukocyte subsets, even in NK cells and macrophages, compared with untreated normal rats. CONCLUSIONS: Data suggest that GnRHa itself did not have an immunomodulating effect when rats were treated for 3 weeks, but that a hypoestrogenic state reduced the size of the ectopic endometrium in association with increases in NK cells and macrophages. Our findings support the hypothesis that ectopic endometrial cells may release immunosuppressive factors.

Animals↗
PubMed1996

Strategic proposals for avoiding toxic interactions with drugs for clinical use during development and after marketing of a new drug--proposals for designing non-clinical and clinical studies--is the non-clinical study useful?

Since sorivudine incident has happened in Japan in 1993, an adverse drug-drug interaction has been of special meanings at each step of new drug development including the discovery step, NDA process, and on market. While it is known that several mechanisms are involved in the drug-drug interactions, the mechanisms related to drug metabolism are 1) inhibition of drug metabolizing enzymes, 2) induction of the enzymes, 3) drug absorption, 4) renal excretion, 5) hepatic transport, and 6) protein binding (displacement) interaction. In this report, proposals to avoid serious/lethal drug-drug interactions are presented with the examples. The proposals are: 1) To estimate the drug-drug interactions in consideration of the mechanisms reported so far, 2) To be especially careful for drugs with a small therapeutic index and severe/lethal toxicity, 3) To estimate the drug-drug interactions in consideration of physiological factors of patients, who receive drugs in combination, 4) Not to leave the mechanism unclear, when some data, which do not deny the critical drug-drug interactions, were obtained, and 5) To conduct the drug-drug interaction studies in humans in careful consideration of the safety.

Animals↗
PubMed1995

Accessibility to tissue-specific genes from methylation profiles of mouse brain genomic DNA.

The DNA methylation status of a large number of genomic loci is visualized simultaneously and quantitatively as two-dimensional gel spots in the newly developed restriction landmark genomic scanning with a methylation-sensitive restriction enzyme (RLGS-M). Here, we demonstrate that RLGS-M using NorI as a methylation-sensitive enzyme could also scan gene loci of mammalian genomes, since almost all of the NotI loci corresponding to randomly chosen RLGS-M spots were located near or in transcriptional units (6 out of 7 NotI-linking clones) when mouse brain genomic DNA was used. This supports the previous prediction that most NotI sites are located in CpG islands (Lindsay and Bird, Nature 1987, 327, 336-338). Furthermore, beginning with RLGS-M spots we examined how to approach their corresponding RNA messages, whose expression may be associated with methylation. We compared RLGS-M patterns among various developmental stages of the mouse brain from embryonic day 9.5 to postnatal 8 weeks or among in vitro cell lines, and detected alterations of RLGS-M spots which were due to methylation of NotI sites. Two experiments using NotI-linking clones or polymerase chain reaction (PCR) were carried out to approach to their corresponding RNA messages. Consequently, we isolated two PCR-amplified clones (# 15 and # 91) which corresponded to methylatable loci and gave positive signals to mRNA from the adult brain. Furthermore, we identified two NotI-linking clones (C211 and C198) whose corresponding NotI loci localized near or at transcriptional units and were methylated in cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Increased neuronal firing in the rat auditory cortex associated with preparatory set.

Extracellular single neuronal firings were recorded in the auditory cortex of rats (n = 4) performing a visual reaction-time task with a warning tone (10 kHz, 10 ms duration), which preceded the imperative light stimulus by an interstimulus interval (ISI) of 1.4 s. Thirty-six neuronal firings were evoked by the warning tone, with the peak latency being between 15 and 55 ms. Among them, nine neurons (25%) showed an increased firing frequency following the evoked response during the ISI, which was, in average, 2.5 times as high as the firing frequency during the baseline period. When the tones were presented independent of the imperative stimulus, such sustained increase in neuronal firing was not observed. Activation of the sensory cortex during the ISI may constitute one of the neuronal modulations related to preparatory set.

Acoustic Stimulation↗
PubMed1995

Effects of metamfetamine on auditory P3 component of event-related potentials in rats.

Three rats were trained with an oddball paradigm, in which 5 kHz frequent and 10 kHz target rare tones were presented with a probability of 0.8 and 0.2. Pressing the lever within 2 s following the onset of the target tones was rewarded with a drop of food paste. Long latency event-related potentials (ERPs) corresponding to those in humans could be recorded in rats. After the administration of metamfetamine (0.2 mg/kg), the amplitude of surface and intracortical N1 increased, indicating an augmentation of the primary cortical response. Surface P3 showed a decrease of amplitude, accompanied by an increase of intracortical negativity. These alterations of ERPs may be caused by the dysregulation of central noradrenergic system.

Amphetamines↗
PubMed1995

Leukocyte subpopulation changes in rats with autotransplanted endometrium and the effect of danazol.

PROBLEM: This study examines immune cell populations in rats with autotransplanted endometrium and determines the effect of danazol on leukocyte subsets. METHODS: As an experimental model of endometriosis, an autologous endometrial segment was implanted in the rat peritoneum. We used flow cytometry to analyze lymphocyte subsets in the peripheral blood (PB) and peritoneal fluid (PF) of the following groups of rats: no treatment, sham operation, endometrial implantation, endometrial implantation treated with danazol, and normal rats treated with danazol. RESULTS: The natural killer (NK) cell population was decreased in both the PB and PF of rats with autotransplanted endometrium. Moreover, NK cells increased in a dose-dependent manner following danazol administration. Surgery itself increased the number of peritoneal macrophages as compared with the untreated group. This elevation was suppressed partially by endometrium-implantation and was attenuated by subsequent administration of danazol in a dose-dependent fashion. CONCLUSIONS: These data suggest that ectopic endometrial cells may release immunosuppressive factors. This is the first documentation that rats with autotransplanted endometrium show the same immunologic changes as humans with endometriosis, and establishes the utility of this model for the study of endometriosis.

Animals↗
PubMed1995

Daily walking combined with diet therapy is a useful means for obese NIDDM patients not only to reduce body weight but also to improve insulin sensitivity.

OBJECTIVE: To evaluate the effects of walking combined with diet therapy (1,000-1,600 kcal/day) on insulin sensitivity in obese non-insulin-dependent diabetes mellitus (NIDDM) patients. RESEARCH DESIGN AND METHODS: Subjects were divided into two groups: 10 patients were managed by diet alone (group D), and 14 patients were placed in the diet and exercise group (group DE). Group DE was instructed to walk at least 10,000 steps/day on a flat field as monitored by pedometer (19,200 +/- 2,100 steps/day), and group D was told to maintain a normal daily routine (4,500 +/- 290 steps/day). A glucose clamp procedure at an insulin infusion rate of 40 microU.min-2.min-1 was performed before and after the 6- to 8-week training program. Mean serum insulin concentrations ranged from 720 to 790 pmol/l. RESULTS: While body weight (BW) in groups D and DE decreased significantly (P < 0.01) during the study, the amount of BW reduction in group DE was greater than that in group D (7.8 +/- 0.8 vs. 4.2 +/- 0.5 kg, P < 0.01). After training, glucose infusion rate (GIR) and metabolic clearance rate (MCR) in group D did not significantly increase; however, GIR and MCR increased significantly in group DE, from 17.21 +/- 1.11 to 26.09 +/- 1.11 mumol.kg-1.min-1 (P < 0.001) and from 3.0 +/- 0.3 to 5.3 +/- 0.4 ml.kg-1.min-1 (P < 0.001), respectively. The analysis of variance showed significant effects of exercise (time x exercise, P = 0.0005) for the improvement of MCR. Significant correlations were also observed between delta MCR and average steps per day (r = 0.7257, P < 0.005) in group DE. CONCLUSIONS: Walking, which can be safely performed and easily incorporated into daily life, can be recommended as an adjunct therapy to diet treatment in obese NIDDM patients, not only for BW reduction, but also for improvement of insulin sensitivity.

Adult↗
PubMed1994

Genomic analysis of a NF1-related pseudogene on human chromosome 21.

A neurofibromatosis type-1 (NF1)-related locus on human chromosome 21 has been characterized. A detailed genomic mapping performed by yeast artificial chromosome (YAC) dot-hybridization revealed that the NF1-related locus is close to the sequence-tagged site (STS), D21S329 (G52E12), which is located on the proximal region of 21q11.2. Sequence analysis showed that this locus seemed to be conserved to the NF1 gene only in several partial regions. Two exon-like segments corresponding to exons 8 and 9 of NF1 were found, in addition to two previously found fragments corresponding to exons 7 and 11. Other exon-like segments were not found in the region so far sequenced. Comparing these homologous segments with the NF1 cDNA, a 2-bp deletion appeared in exon 8 of the locus, resulting in the existence of stop codons in all reading frames. In addition, there were no amplified fragments derived from the related locus by reverse transcription-PCR. Thus, our results suggest that the NF1-related locus on chromosome 21 is a nonprocessed pseudogene.

Animals↗
PubMed1994

beta 2-GPI-dependent and independent binding of anticardiolipin antibodies in patients with recurrent spontaneous abortions.

We measured anticardiolipin antibodies (aCL) in plasma samples from 214 women with a history of recurrent spontaneous abortions by an enzyme-linked immunosorbent assay (ELISA) utilizing solid phase cardiolipin (CL) and beta 2-glycoprotein I (beta 2-GPI). Sixteen patients (7.5%) were positive for beta 2-GPI-dependent aCL. Though beta 2-GPI appeared to enhance the binding of aCL, beta 2-GPI-independent aCL were also observed in these patients (4.7%). The patients were classified into three groups on the basis of their medical history, and analysis of data of individual groups revealed that the incidence of beta 2-GPI-dependent aCL was significantly higher in patients who had experienced at least one fetal loss in the second or third trimester.

Abortion, Habitual↗
PubMed1994

Event-related potentials during an auditory discrimination task in rats.

Event-related potentials (ERPs) during an auditory discrimination task were recorded both on the surface and at a depth of the auditory cortex in rats. Ten-kilohertz rare and 5-kHz frequent tones were used with the probabilities of 0.2 and 0.8. Lever pressing within 2 s, following the onset of the rare tone, was rewarded with food paste. In the performing condition, the surface ERPs for the rare tones consisted of P30, N50, P80, N130, and P290 components. Only the surface P30 and N50 showed a polarity reversal at the intracortical electrode, suggesting that these components are generated in the auditory cortex. The P290 was elicited in the performing condition but not in the resting condition, showing its task-relevancy. These results suggest that auditory ERPs similar to those in humans can be recorded in rats during an active discrimination task.

Acoustic Stimulation↗
PubMed1994

Possible noradrenergic dysfunction in schizophrenia.

In spite of extensive studies over the last 2 decades to find direct evidence in support of the dopamine hypothesis of schizophrenia, no undisputed experimental data has been obtained. In contrast, estimation of noradrenalin (another major catecholamine) and its metabolites in postmortem brain and in the cerebrospinal fluid appears to be producing consistent results. To understand the meaning of this change for the pathogenesis of the illness, we have carried out animal experiments in which reproducibility of schizophrenic signs and symptoms by noradrenergic dysfunction, and treatability of the disorder by modulation of noradrenergic activity were studied. First, psychophysiological signs in skin conductance responsiveness (nonhabituating or nonresponding change) and smooth pursuit eye movement (spiky or stepwise pursuit) could be reproduced by enhancing or suppressing central noradrenergic activity. Behavioral abnormalities resembling schizophrenic symptoms are known to be reproducible by over- or underactivity of the system (overarousal or underarousal syndrome). Secondly, the action of various drugs capable of modulating schizophrenic symptoms was analyzed in relation to noradrenergic activity. Haloperidol, in particular, had a potent suppressing effect on skin conductance activity (spontaneous fluctuation rate and habituation rate) when administered chronically, suggesting its inhibitory action on noradrenergic activity.

Animals↗
PubMed1994

Functional influence of the central noradrenergic system on the skin conductance activity in rats.

Pharmacological studies on neuroleptics and amphetamine strongly suggest that some dysfunction of the central catecholamine system may play a key role in the pathogenesis of schizophrenia. Our previous studies have demonstrated that intraventricular administration of 6-hydroxydopamine, a selective neurotoxin of the catecholamine neuron, can reproduce schizophrenia-like abnormalities in the skin conductance activity. In the present experiments, effects of pharmacological modulation of the central noradrenergic activity were studied in rats. Stimulation of the central noradrenergic activity by yohimbine (0.6 mg/kg, i.m.) slowed down the habituation of the skin conductance response (SCR) and increase the spontaneous fluctuation of the skin conductance (SF), while inhibition of the activity by clonidine (0.06 mg/kg, i.m.) accelerated or obliterated the SCR and decreased the SF frequency. If the functional significance of the central noradrenergic system lies in vigilance control, the present results are consistent with classical theory in psychophysiology: the habituation rate of SCR and the frequency of SF are correlated well with each other and both indices reflect arousal level. The disorder of the system should produce not only these psychophysiological abnormalities but also psychological disturbances; i.e., overarousal and underarousal syndromes. Therefore, the dysfunction of the noradrenergic system might constitute an essential aspect of schizophrenic disorder.

Acoustic Stimulation↗
PubMed1994

A PCR-mediated method for cloning spot DNA on restriction landmark genomic scanning (RLGS) gel.

A PCR-mediated direct cloning for target spot DNA from RLGS gel has been established. The method consists of PCR amplification of adaptor-ligated spot DNA fragments without excluding similar-sized DNA fragments co-localized on RLGS gel, and following selective ligation with the NotI-dT vector. Applying this method, we have successfully cloned several DNA fragments derived from target spots whose intensities change developmentally due to DNA methylation in the telencephalon of C3H/HeN mice. Since only a few micrograms of total DNA is sufficient for our spot cloning, our method may be highly useful when the total DNA sample prepared for cloning is limited.

Animals↗
PubMed1994

The influence of intravenous immunoglobulin treatment on maternal immunity in women with unexplained recurrent miscarriage.

PROBLEM: Recently the protective value of high-dose intravenous immunoglobulin (IVIG) in the treatment of unexplained recurrent miscarriage has been reported to be similar to that of conventional immunotherapy with paternal leukocytes. We examined the effect of IVIG treatment on the cellular and humoral level of maternal immunity to demonstrate the possible mechanism by which IVIG might act to prevent recurrence of pregnancy loss. METHOD: Eight patients were treated with a 20- to 25-g dose of IVIG every 2 to 3 wk during their first-trimester pregnancies. The development of anti-idiotypic autoantibodies against maternal T-cell receptors, maternal anti-paternal lymphocyte antibodies detected by flow cytometric crossmatch, and changes of maternal lymphocyte subpopulations were monitored before pregnancy and then weekly during IVIG treatment. RESULTS: Five of eight patients gave birth successfully after IVIG treatment given during the first trimester of pregnancy (success rate: 62.5%). Although we could not demonstrate a general immunological effect of IVIG on maternal immunity in vivo, a few significant changes of immunological parameters were found in some patients. CONCLUSION: Our results suggest that the effect of IVIG on maternal immunity is not a passive increase of blocking antibody including anti-HLA antibody or modification of maternal T-cell subsets but, more likely, a passive increase of anti-idiotypic antibody against anti-HLA antibody or soluble HLA antigens. However, whether the immunomodulating effect of IVIG is related to its possible mechanism to prevent abortion remains unestablished.

Abortion, Habitual↗
PubMed1994

Comparison of DNA methylation patterns among mouse cell lines by restriction landmark genomic scanning.

Restriction landmark genomic scanning (RLGS) is a novel method which enables us to simultaneously visualize a large number of loci as two-dimensional gel spots. By this method, the status of DNA methylation can efficiently be determined by monitoring the appearance or disappearance of spots by using a methylation-sensitive restriction enzyme. In the present study, using RLGS with NotI, we examined, in comparison with a brain RLGS profile, the status of DNA methylation of more than 900 loci among three types of mouse cell lines: the embryonal carcinoma cell line P19, the stable mesenchymal cell line 10T1/2, and our established neuroepithelial (EM) cell lines. We found that the relative numbers of RLGS spots which appeared were less than 3.3% of those surveyed in all cell lines examined. However, 5 to 14% of spots disappeared, the numbers increasing with an increase in the length of the culture period, and many spots were commonly lost in 10T1/2 and in three EM cell lines. Thus, for these cell lines, many more spots disappeared than appeared. However, the numbers of spots disappearing and appearing were well balanced, and the ratio in P19 cells was almost equal to that in liver cells in vivo. These RLGS experimental observations suggested that permanent cell lines such as 10T1/2 are hypermethylated and that our newly established EM cell lines are also becoming heavily methylated at common loci. On the other hand, methylation and demethylation seem to be balanced in P19 cells in a manner similar to that in in vivo liver tissue.

Aneuploidy↗