Biomedical subjects

M Kato

Publications and source records attributed to M Kato.

At least 667 records · Page 37Linked to original sources

PubMed1998

Large scale isolation of osteoclast-specific genes by an improved method involving the preparation of a subtracted cDNA library.

BACKGROUND: Osteoclasts play crucial roles in bone resorption, which triggers bone remodeling. Molecular mechanisms underlying these osteoclast-specific biological functions remain elusive because only a limited number of osteoclast-specific genes have been identified. To circumvent this, we isolated a large number of osteoclast-specific genes by preparing a subtracted cDNA library of high quality. RESULTS: We first constructed a plasmid expression vector (pAP3neo) that allowed an efficient subtraction. Then, we improved the standard protocols for preparation of the cDNA library and the subsequent subtraction procedure. Using our protocol, we prepared a rabbit osteoclast cDNA library of high complexity. Subsequently, we prepared an osteoclast-specific cDNA library of high complexity by subtracting it with biotin-labelled mRNA, derived from rabbit spleen through the biotin-avidin method. The resulting library included a high proportion of full-length cDNA inserts. Using DNA dot blot analysis, we found that the osteoclast-specific cDNA clones were highly enriched in this subtracted cDNA library, i.e. nearly 70% of the analysed clones were primarily detected in osteoclasts but not in spleen. Multiple-tissue Northern analysis also showed that many of these clones were expressed almost exclusively in osteoclasts. DNA sequencing of randomly selected clones showed that 424 cDNA species out of 1136 analysed were novel. DNA sequencing also showed that our subtracted cDNA library was almost equalized, suggesting that the library may contain almost all of the osteoclast-specific genes. CONCLUSION: From these data, we conclude that our subtraction protocols, and the subsequent procedure for the analysis of the isolated clones developed here, are useful for the comprehensive isolation and identification of transcriptionally up- or down-regulated genes in general.

Animals↗

PubMed1998

A polysomnographic study of sleep patterns in normal humans with low- or high-anxiety personality traits.

To clarify the effects of anxiety-related personality traits on sleep patterns, polysomnographic examinations (PSG) were performed over 4 consecutive nights on normal humans who tested within the low- or high-anxiety ranges. The subjects consisted of two groups of six male university students who scored either less than 45 points (low-anxiety group) or more than 55 points (high-anxiety group) on the Spielberger's State Trait Anxiety Inventory. Compared to the levels of sleep change in the high-anxiety group, the low-anxiety group exhibited a greater change in REM sleep and stage 2 sleep. The REM sleep in the low-anxiety group was shorter on the first and second nights compared to the third and fourth nights, and the stage 2 sleep was longer on the first night than on the remaining three nights. Thus, the low-anxiety group showed a first-night effect followed by partial recovery on the second night, while the high-anxiety group exhibited no obvious first-night effect. These results suggest that there is a difference in sleep patterns, assessed by consecutive PSG, between those with low- and high-anxiety traits, and that anxiety-related personality traits attenuate the occurrence of the first-night effect, reflecting a lower adaptability to a novel environment.

Adult↗

PubMed1998

The herbal medicine Sho-saiko-to inhibits growth and metastasis of malignant melanoma primarily developed in ret-transgenic mice.

Sho-saiko-to is the most popular herbal medicine in Japan. We investigated the anti-tumor and anti-metastatic effects of Sho-saiko-to and its chemically defined ingredients on the primary skin melanoma that developed in a metallothionein-I (MT)/ret transgenic mouse line and on a melanoma cell line (Mel-ret), which was derived from a primary tumor developed in a MT/ret transgenic mouse. In vitro, Sho-saiko-to suppressed the growth of Mel-ret cells more strongly than any single ingredient of Sho-saiko-to, although baicalin as one of several ingredients tested also suppressed it significantly. In vivo, Sho-saiko-to (i) significantly (p < 0.02) prolonged the onset of tumor development (1.5 mo), (ii) definitely retarded the transition to malignancy, (iii) significantly decreased the incidence of distant metastasis to brain (p < 0.002), kidney (p < 0.05), and liver (p < 0.05) at the malignant stage, and (iv) significantly (p < 0.02) prolonged life span (2.6 mo). Moreover, Sho-saiko-to and baicalin down-regulated the matrix metalloproteinase-2 and -9 expression levels, and upregulated their inhibitor expression level in both the primary tumors and Mel-ret cells. In conclusion, Sho-saiko-to displayed anti-tumor and anti-metastatic effects on melanoma with regulation of the balance of matrix metalloproteinase and tissue inhibitor of the matrix metalloproteinase levels.

Animals↗

PubMed1998

Highly sensitive urine-based enzyme-linked immunosorbent assay for detection of antibody to Helicobacter pylori.

BACKGROUND: Enzyme-linked immunosorbent assay (ELISA) has been widely used for detection of Helicobacter pylori (H. pylori), but sample collection is often invasive, complicated, and expensive. Urine samples can be obtained noninvasively and are easier and safer to handle than serum samples. A urine-based ELISA, if found to be accurate, would therefore be a useful alternative to serum-based tests for H. pylori. METHODS: An ELISA method was developed for detection of antibodies to H. pylori in urine. Its sensitivity and specificity were compared with those of three commercially available serum-based ELISA kits and the 13C urea breath test (13C-UBT) using samples from 99 healthy volunteers and 20 patients with gastric disorders. RESULTS: With the assumption that 13C-UBT results are 100% accurate, the sensitivity and specificity of the urinary ELISA were 99% and 100%, respectively, and the accuracy (99%) was superior to those of the three serum ELISAs tested. Immunostaining profiles on Western blot analysis using serum samples were almost identical to those obtained using paired urine samples. CONCLUSIONS: These findings suggest that the differences observed among ELISA test results may be due principally to differences between the profiles of antigen coated on plates for the assays, rather than to differences between antibodies in serum and urine. The urine-based ELISA (URINELISA H. pylori) developed in this study is very accurate and would be useful for screening H. pylori infection as an alternative to serum ELISAs.

Antibodies, Bacterial↗

PubMed1998

The UBiT-100 13CO2 infrared analyzer: comparison between infrared spectrometric analysis and mass spectrometric analysis.

BACKGROUND: Although the 13C-urea breath test is the most accurate noninvasive method for detecting the presence of H. pylori infection, the requirement for an expensive mass spectrometer to analyze breath samples has prevented physicians from providing rapid results near the patient. Recently, a new type of infrared spectrometric analyzer, the UBiT-100, was developed for analyzing 13CO2 in breath. The purpose of this study is to compare results analyzed by the UBiT-100 to those analyzed by the mass spectrometric method. METHODS: Four hundred and fifty-three subjects participated in this study. Breath samples were collected before administration of 100 mg of 13C-urea and at 10, 20, 30, 45 and 60 min after administration. Subjects were asked to hold their breath for 10 sec and then exhale in order to collect breath samples containing more than a 2% concentration of CO2. Samples were then analyzed by both methods. RESULTS: The correlation analysis using values at 20 min after the administration of the study drug (433 points) was excellent with the regression equation of Y = 1.034x - 0.203; r = .996. The results of the UBiT-100 were available in 6 min, making the entire testing procedure less than 30 min. CONCLUSIONS: The UBiT-100 infrared analyzer provides a simple and accurate method of performing the urea breath test while the patient is still in the doctor's office.

Breath Tests↗

PubMed1998

The Effects of Local Cooling on Thermophysiological Response in Participants Wearing Dust-Free Garments.

This study was designed to find the effects of two kinds of dust-free garments with (A) and without (B) frozen gel strip (FGS), and half-naked clothing (brassiere and shorts; C) on thermophysiological parameters and on temperature and humidity within clothing. The heart rate, rectal, and skin temperatures as well as sweat rate and clothing microclimate were measured during 140 min in 9 healthy females. Inquiries were also made into the subjective rating of thermal, humidity, and comfort sensations. The main findings in our experiments are as follows: (a) Physiological parameters such as rectal and skin temperatures (chest and forehead), heart rate, and sweat rate were clearly lowest in garb C, intermediate in garb A, and highest in garb B throughout the experiment; (b) Temperature and humidity within clothing were lower in garb A than in garb B; (c) More than half of the 9 participants decreased thermal sensation by wearing garb A. These results suggest that the usage of FGS could improve the heat load in lightly working participants wearing dust-free garments.

Journal Article↗

PubMed1998

Response to platelet-derived growth factor by phenotypically different cultured human aortic smooth muscle cells.

We investigated the effects of PDGF on DNA synthesis and mitogen-activated protein (MAP) kinase activity, and demonstrated that the adult intimal SMC was concentration-dependently stimulated by all PDGF isoforms in terms of both [3H]thymidine incorporation and MAP kinase activation, with PDGF-BB and -AB being more potent than PDGF-AA. The intimal SMCs and the neonatal SMCs showed a similar response with regard to MAP kinase activation. On the other hand, the intimal SMCs expressed many more PDGF receptors than the adult medial SMCs, which expressed a greater amount of PDGF-A chain mRNA and showed a lesser response to PDGFs. These results suggest that the intimal SMCs have a relatively high potential to react to exogenous PDGFs, whereas the adult medial SMCs depend on endogenous or autocrine secretion of PDGF-AA.

Adult↗

PubMed1998

Human leukotriene B4 omega-hydroxylase (CYP4F3) gene: molecular cloning and chromosomal localization.

Leukotriene B4 (LTB4) omega-hydroxylase catalyzes the conversion of LTB4 into a biologically less active product, 20-hydroxy-LTB4. In a preceding paper (Kikuta et al., 1993), we showed human polymorphonuclear leukocyte (PMN) LTB4 omega-hydroxylase to be a novel form of cytochrome P450, designated CYP4F3, on the basis of its cDNA cloning and expression in yeast cells. Here, we have isolated the gene encoding CYP4F3 and determined its genomic organization and chromosomal localization. The CYP4F3 gene contained 13 exons and spanned approximately 22.2 kb. The cDNA of CYP4F3 contained 5050 nucleotides excluding the poly(A) tail. The translation initiation codon (ATG) was present in exon II. Primer extension and S1 mapping analyses indicated that the transcription initiation site is 49 nucleotides upstream from the 3' end of exon I, and no other initiation sites were detected. A TATA-box-like sequence (TACAT) and 120-b GC-rich sequence were observed just before transcription initiation site. Several putative regulating elements recognized by the GATA family, MZF-1, CACCC binding protein, and C/EBP, were identified in its 5' flanking region. Genomic DNA screening for CYP4F3 and Southern blot analysis suggested the existence of other CYP4F genes in addition to CYP4F3 and CYP4F2 in the human genome. Fluorescence in situ hybridization demonstrated that the CYP4F3 gene is located at 19p13.2.

Amino Acid Sequence↗

PubMed1998

Decreased mitogen response of splenic lymphocytes in obese Zucker rats is associated with the decreased expression of glucose transporter 1 (GLUT-1).

We reported previously that obesity is a risk factor for deteriorating cellular immune functions in aging. However, the mechanism by which obesity decreases cellular immunity remains to be elucidated. To determine the mechanism of the decrease in cellular immunity with obesity, lean (Fa/?) and obese (fa/fa) 12-mo-old Zucker rats were used. The mitogen response of splenic lymphocytes in obese Zucker rats was significantly lower than that of lean Zucker rats, which was not restored by in vitro treatment with indomethacin (10 micromol/L), an inhibitor of prostaglandin E2 (PGE2). In addition, PGE2 production by splenic lymphocytes was not greater in obese than in lean Zucker rats. Glucose consumption by splenic lymphocytes after in vitro incubation with concanavalin A (conA) for 48 h was also significantly lower in obese Zucker rats. Expression of glucose transporter 1 (GLUT-1), analyzed by Western blot analysis, was lower in splenic lymphocytes of obese than in lean Zucker rats. However, the expression of the conA receptor in splenic lymphocytes, analyzed by flow cytometry with fluorescein isothiocyanate-conjugated conA, was not significantly different between lean and obese Zucker rats. In conclusion, the decreased mitogen response of splenic lymphocytes in obese Zucker rats may be due in part to the decreased uptake of glucose as the main energy source for lymphocytes at the stage of proliferation and may be associated with the decreased expression of GLUT-1.

Animals↗

PubMed1998

Nafamostat mesilate, a kallikrein inhibitor, prevents pain on injection with propofol.

We have examined the preventative effect of nafamostat mesilate, a kallikrein inhibitor, on pain on injection with propofol in a randomized, double-blind study. A control group (n = 110) and a nafamostat (n = 103) group received 5% glucose 0.02 ml kg-1 and nafamostat 0.02 mg kg-1 diluted with 5% glucose, respectively, followed 1 min later by 1% propofol injected at a rate of 200 mg min-1. Pain scores recorded during injection of propofol were significantly less in the nafamostat than in the control group. In another 10 patients, blood concentrations of nafamostat were measured after administration of nafamostat 0.02 mg kg-1 i.v. Mean nafamostat concentration 1 min after injection was 0.1 (SD 0.05) mumol litre-1, which is sufficient to inhibit plasma kallikrein activity. We conclude that pretreatment with nafamostat 0.02 mg kg-1 significantly reduced pain on propofol injection and this effect may be caused by a reduction in kallikrein activity.

Adolescent↗

PubMed1998

Prospective and retrospective studies of recovery in aphasia. Changes in cerebral blood flow and language functions.

Prospective and retrospective language evaluations and single photon emission computed tomography (SPECT) scans were performed in order to study the relationship between post-stroke recovery from aphasia and changes in cerebral blood flow (CBF) in groups of patients who had made a good recovery and those who had not. For the prospective study, 20 right-handed patients with aphasia secondary to an acute cerebrovascular accident (CVA) in the left middle cerebral artery territory received language evaluations with a Japanese Standard Language Test of Aphasia (SLTA), and SPECT scans performed twice, at a mean of 3.2 and a mean of 9.2 months post-onset. Only one slice of SPECT data was analysed. A significant correlation was observed between the severity of the initial language deficit and initial CBF on the left side, but not the right. Initial CBF was not a predictor for future language recovery in either hemisphere. There was a correlation between the change in the left mean hemispheric CBF (but not the right) and the change in the overall SLTA severity rating from 3 to 9 months post-stroke. In the retrospective study, 16 right-handed patients with residual aphasia secondary to CVA in the left middle cerebral artery territory received SLTA and SPECT at a mean of 82.8 months post-onset. The patients had also received initial language evaluation with SLTA at a mean of 6.5 months post-onset. In contrast to the prospective study, the results demonstrated that the mean left hemispheric CBF at approximately 7 years post-onset did not differ between good and poor recovery groups. However, the mean right hemispheric CBF of the good recovery group was higher than that of the poor recovery group in the frontal and the thalamic regions, and also in the left frontal region. The results of these complementary studies suggest that the initial language recovery within the first year post-onset may be linked primarily to functional recovery in the dominant hemisphere, where an increase in CBF was observed at 9 months post-onset. The increased perfusion adjacent to the lesion may be crucial for early recovery in aphasia. Subsequent language recovery and the long-term recovery in aphasia may be related to slow and gradual compensatory functions in the contralateral hemisphere, specifically in the homotopic frontal and thalamic areas.

Adult↗

PubMed1998

The roles of individual cysteine residues of Sendai virus fusion protein in intracellular transport.

The role of intramolecular disulfide bonds in the fusion (F) protein of Sendai virus was studied. The 10 cysteine residues were changed to serine residues using site-directed mutagenesis. None of the cysteine mutant F proteins reacted with a monoclonal antibody specific for the mature conformation of the F protein, but eight of ten mutants reacted with an immature conformation-specific monoclonal antibody. The transport of these mutant proteins to the cell surface was drastically reduced. All of the cysteine mutant F proteins remained sensitive to endoglycosidase H (endo H) for 3 h after their synthesis. Moreover, cell surface transport of the hemagglutinin-neuraminidase (HN) protein co-expressed with each of these cysteine mutant F proteins was also reduced. These results suggest that all cysteine residues participate in the formation of intramolecular disulfide bonds, that co-translational disulfide bond formation is crucial to the correct folding and intracellular transport of the F protein, and that interaction of the F and HN proteins takes place intracellulary.

Animals↗

PubMed1998

Reactivity of soybean lipoxygenase-1 to linoleic acid entrapped in phosphatidylcholine vesicles.

The linoleic acids embedded in the SUVs of soy-PC, DMPC, and DPPC served as substrate for soybean lipoxygenase-1 (L-1). The initial velocity of the catalytic reaction and the concentration of the substrate showed a hyperbolic relation. The Km values of L-1 for the linoleic acids in soy-PC, DMPC, and DPPC vesicles were 0.07, 0.09, and 0.11 mM, respectively, being comparable with that for Tween-20 micellar linoleic acid. Soy-PC and DMPC competitively inhibited the enzyme with Ki values of 0.20 and 0.13 mM, respectively, whereas DPPC had no effect. DSC analysis revealed the phase separation of linoleic acid and DPPC in vesicles in the temperature range in which the enzyme reaction was carried out. This may account for the lack of inhibitory effect of DPPC on the enzyme. From the temperature dependence of the specific activity of the enzyme, the Ea values of the catalytic reaction were estimated to be 26.7 and 35.3 kJ.mol-1 for soy-PC and DPPC vesicles, respectively. For linoleic acid-DMPC vesicles, a two-phase temperature dependence of the activity across the transition temperature of the mixed vesicles was suggested.

Calorimetry, Differential Scanning↗

PubMed1998

Review article: Long-term Helicobacter pylori infection--from gastritis to gastric cancer.

An analysis carried out in 1994 by the WHO International Agency for Research on Cancer (IARC) resulted in Helicobacter pylori being designated as a Group 1 carcinogen and thus clearly having an association with the development of gastric cancer. In the case of H. pylori, the evaluation was made solely on the basis of epidemiological results. In Japan, in 1993, only 235,000 of the 60 million people with H. pylori had gastric cancer. This represents only 0.4% of the infected population. Each individual reacts in a unique way to H. pylori infection in terms of the inflammatory response. The probability of developing cancer will be determined by environmental factors such as diet, duration of or age at acquisition of H. pylori infection, the virulence of H. pylori strains, and host factors including genetic make-up.

Antibodies, Bacterial↗

PubMed1998

Augmented exercise plasma noradrenaline with impaired chronotropic responsiveness in patients with hypertrophic cardiomyopathy.

1. There is controversy regarding plasma catecholamine levels in patients with hypertrophic cardiomyopathy (HCM) and few data exist on serial plasma catecholamine measurements during exercise. The present study determined whether cardiovascular and plasma catecholamine responses to exercise were altered in patients with HCM. 2. Plasma noradrenaline (NA) and adrenaline were measured at rest, at the end of each stage during exercise and immediately and 5 min after submaximal treadmill exercise in 15 patients with non-obstructive HCM (13 males, two females; mean (+/- SEM) age 54 +/- 3 years) and in 15 age- and sex-matched controls. The ratio of the increment in heart rate (HR) divided by the increment in plasma NA during exercise (delta HR/delta NA) was used as an index of chronotropic sympathetic responsiveness to exercise. 3. Exercise duration was shorter (11.2 +/- 0.6 vs 8.7 +/- 0.6 min for control vs HCM, respectively; P < 0.01) and diastolic blood pressure was significantly higher at stages I and II of modified Bruce protocol HCM. 4. Resting plasma NA levels (149 +/- 17 vs 167 +/- 28 pg/mL for control vs HCM, respectively; NS) were not different, but plasma NA levels at stages I and II were significantly higher in HCM than in controls (243 +/- 26 vs 399 +/- 69 pg/mL (P < 0.05) and 308 +/- 30 vs 548 +/- 110 pg/mL (P < 0.05), respectively). 5. Peak plasma NA levels were not significantly higher in HCM than in controls (578 +/- 59 vs 918 +/- 184 pg/mL, respectively; NS). 6. The ratio delta HR/delta NA was significantly lower in HCM compared with control at stages I and II (0.49 +/- 0.10 vs 0.21 +/- 0.05 (P < 0.05) and 0.38 +/- 0.06 vs 0.20 +/- 0.05 (P < 0.05), respectively). There were no differences in plasma adrenaline responses during exercise between the two groups. 7. Patients with HCM had augmented plasma NA levels during submaximal exercise with a higher diastolic blood pressure response. Chronotropic sympathetic responsiveness was impaired during the early stages of exercise in patients with HCM.

Adult↗

PubMed1998

Polysomnography and body temperature changes by phototherapy in a delayed sleep phase syndrome case.

We examined polysomnography (PSG) and body temperature in a patient with delayed sleep phase syndrome who responded to phototherapy. The patient was a 31-year-old woman whose condition had slightly improved by a vitamin B12 administration. Phototherapy was administered to her in combination with the vitamin B12 medication, and this combined treatment successfully advanced her delayed sleep phase. On PSG, the regimen showed shortened sleep latency, decreased total sleep time and stages 1 and 2 sleep, and increased slow wave sleep. Phototherapy also improved temporal distribution of delta half-waves (0.5-2.0 Hz, > or = 31 microV) as well as phase relationship between sleep and body temperature.

Adult↗

PubMed1998

Melatonin treatment for rhythm disorder.

We tried melatonin treatment in two patients with non-24 h sleep-wake syndrome, who did not respond to treatments by vitamin B12, bright light therapy, or hypnotics. In one patient, melatonin 5-10 mg improved difficulty in falling asleep and in waking, although it failed to improve the sleep-wake rhythm. In another patient, melatonin 3 mg successfully changed the sleep-wake rhythm from free-running pattern to delayed sleep phase pattern. However, melatonin re-administration after a 4-month drug-free interval failed to improve his free-running sleep-wake rhythm. These results suggest that melatonin acted as a sleep inducer in one patient and as a phase setter in the other, although the effect on the latter patient was transient.

Adult↗

PubMed1998

Evaluation of the prognostic significance of microvessel count and tumor size in renal cell carcinoma.

BACKGROUND: We analyzed the significance of tumor microvessel count and tumor size as prognostic factors for renal cell carcinoma (RCC). METHODS: T criteria (local invasion), M criteria (distant metastasis), macrovenous invasion, histological grade, cell type, tumor size, and tumor microvessel count were investigated as prognostic factors in 62 patients with RCC. Microvessel density was evaluated by immunohistochemical staining of the endothelial cells. RESULTS: No significant correlation was found between tumor size and the other prognostic factors. Analysis of the survival rate based on tumor size revealed that patients with a small tumor tended to show a good prognosis, but no significant difference was found in the prognosis between medium-sized and large-sized tumor groups. Analysis of all patients revealed that M criteria, macrovenous invasion, cell type and microvessel count were significant factors and that in 37 patients with low-stage tumors, the only significant prognostic factor was microvessel count (P=0.003). CONCLUSION: Tumor size cannot be readily justified as a determinant of prognosis in patients with medium- or large-sized tumors. In patients with low-stage tumors, microvessel count served as the only significant determinant for prognosis. When the patients were considered as a whole, microvessel count, M criteria, macrovenous invasion and histological grade were significant prognostic factors.

Adult↗