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Biomedical subjects

Y Miyake

Publications and source records attributed to Y Miyake.

At least 55 records · Page 3Linked to original sources

A field evaluation of in situ biodegradation of trichloroethylene through methane injection.

A field study of biodegradation of trichloroethylene (TCE) through methane injection was conducted at the yard of a home in Japan. Methane was selected as the safest substrate for injection into groundwater. Methane, oxygen, nitrate, and phosphate were introduced into groundwater contaminated with 220 microg/L of TCE. After a week of biostimulation, methane concentrations gradually decreased below the detection limit. Methane oxidizing bacterial numbers increased from 10 to 10(4) cells/mL with methane consumptions. During methane injection. 10-20% of TCE removal was observed. The biotransformation yield was 3-13 mgTCE/gCH4 in this field test. After methane injections were stopped, TCE removal was not observed. These results indicated that bioremediation using methane was useful as a safe technology for a TCE-contaminated area near homes.

Biodegradation, Environmental↗

Unilateral cone dysfunction with bull's eye maculopathy.

OBJECTIVE: To report a Japanese subject who presented with an acute-onset, unilateral cone dysfunction with bull's eye maculopathy and to describe the functional changes determined by psychophysical and electrophysiologic tests. DESIGN: A single observational case report. METHODS: In addition to a complete ophthalmic examination, the subject underwent some electrophysiologic and psychophysical tests. MAIN OUTCOME MEASURES: Kinetic visual field test, cone and rod perimetry (two-color perimetry), full-field electroretinograms (ERGs), focal macular ERGs, and multifocal ERGs. RESULTS: The full-field ERGs and two-color perimetry showed a predominant loss of cone function in the right eye, whereas the left eye was normal. Cone perimetry and multifocal ERGs revealed that there were small regions functioning normally and other retinal areas that were severely altered in the right eye. CONCLUSIONS: The topographical function analysis suggested that the disorder affected the retina unevenly. The cause of this rare case of unilateral cone dysfunction with bull's eye maculopathy still remains unknown.

Acute Disease↗

A simple loop method for the automated preparation of (11C)raclopride from (11C)methyl triflate.

A simple automated preparation of [11C]raclopride by reaction of [11C]methyl triflate with demethylraclopride triflate is described. The conventional bubbling of [11C]methyl triflate into the precursor solution was compared with two alternative methods which used a commercially available C18 cartridge (on-column method) or an empty PTFE tube (loop method) as support for the precursor solution. The influence of several solvents was assessed for all three methods. The on-column method showed excellent trapping efficiencies of [11C]methyl triflate but gave the lowest radiochemical yields. The loop method proved to be a simplified alternative to the bubbling method, giving comparable radiochemical yields with less precursor and offering an easy way to transfer the reaction mixture into an HPLC column. By the simple-loop method [11C]raclopride could be prepared in over 40% radiochemical yields (decay-corrected and based on [11C]methyl triflate).

Carbon Radioisotopes↗

Bone marrow micrometastases detected by RT-PCR for mammaglobin can be an alternative prognostic factor of breast cancer.

PURPOSE: To evaluate a new prognostic factor of breast cancer, bone marrow micrometastases which was detected by RT-PCR for mammaglobin, a sensitive molecular marker of breast cancer, was examined. MATERIALS AND METHODS: One hundred and eleven samples from stage I-III breast cancer patients were examined. Bone marrow micrometastases and clinicopathological parameters, which were age, tumor size, lymph node metastasis and status of the estrogen receptor, were evaluated for the prognostic factor by statistical analysis. RESULTS: Median follow-up time was 21.1 months. Thirty-three (29.7%) out of 111 samples were RT-PCR positive. Eight cases (24.2%) in this group showed recurrent lesions in the distant organs. Whereas six (7.7%) out of 78 RT-PCR negative patients had distant recurrences. In the premenoposal patients, and in the patients with axillary lymph node metastases, RT-PCR positive cases showed significantly higher distant recurrent rate. Bone marrow micrometastases, axillary nodal status, and estrogen receptor were independent prognostic factors for breast cancer by both univariate and multivariate analysis. CONCLUSIONS: Bone marrow micrometastases detected by RT-PCR for mammaglobin can be a useful predictive marker for early distant recurrence of breast cancer.

Adult↗

Differential daily expression of Per1 and Per2 mRNA in the suprachiasmatic nucleus of fetal and early postnatal mice.

It is well known that there are circadian rhythms of 2-deoxyglucose uptake and neuronal firing in the rat suprachiasmatic nucleus (SCN) during fetal and early postnatal periods. A core clock mechanism in the mouse SCN appears to involve a transcriptional feedback loop in which CLOCK and BMAL1 function as positive regulators and three mPeriod (mPer) genes play a role in negative feedback. Per genes expression occurs not only in the adult SCN but also in the fetal SCN. However, the developmental change in these genes remains unclear. In this experiment, we examined the day--night pattern of expression of Per1 and Per2 mRNA in the mouse SCN and cerebral cortex on embryonic day 17, postnatal day 3, and in young adult mice under a light-dark cycle. Daily rhythms of mRNA content were observed in mPer1 but not mPer2 in the fetal SCN. Interestingly, the expression of mPer2 in the SCN was high throughout the entire day, and a significant daily rhythm of this gene was observed on postnatal day 6. The expression pattern of SCN mPer1 in constant darkness was similar to that seen in the light-dark cycle. The present results suggest that the daily oscillation of mPer1 but not of mPer2 in the SCN in fetal and early postnatal mice may be associated with the daily rhythms of 2-deoxyglucose uptake and neuronal firing.

ARNTL Transcription Factors↗

Laparoscopic findings in primary sclerosing cholangitis.

BACKGROUND AND STUDY AIMS: Primary sclerosing cholangitis (PSC) is a cholestatic disease characterized by segmental narrowing and dilatation of bile ducts. Few studies have been performed on the laparoscopic findings associated with this disease, and the present study was intended to assess the usefulness of laparoscopy for the diagnosis and staging of PSC. PATIENTS AND METHODS: Six patients were examined by laparoscopy with liver biopsy. Repeated laparoscopy was performed in three patients. RESULTS: Laparoscopy revealed coarse surface irregularity and discoloration. Surface irregularity was classified into four grades: smooth, shallowly depressed, undulated, and nodular. The affected area showed whitish yellow discoloration. The discolored area was demonstrated as a poorly stained area by intravenous injection of indocyanine green (ICG). Lobular markings became apparent because of the yellow color change in the portal tract, resulting in a leopardskin-like appearance. Lymph-vessel dilatation was seen in advanced stages. Repeated laparoscopy of a patient without treatment demonstrated a progression from a smooth surface to a shallow depression with leopardskin-like markings. On the other hand, the two patients treated with immunosuppressive agents showed improvement of liver swelling and disappearance of the leopardskin-like markings and lymph-vessel dilatation. CONCLUSIONS: Laparoscopy may provide useful information for the diagnosis and staging of PSC.

Adolescent↗

A splicing factor, Prp8: preferential localization in the testis and ovary in adult mice.

Prp8 is a splicing factor of 220 kDa originally identified in yeast and is a component of the U5 small nuclear ribonucleoprotein particle. Mouse Prp8 cDNA was cloned and shown to share 62.6 and 68.2% sequence identity with the yeast homologue at the amino acid and nucleotide level, respectively, while it differs by only 3 amino acid residues from the human homologue. During mouse embryogenesis, Prp8 is expressed intensely at day 9.5 of gestation, and its expression decreases progressively during embryogenesis. In adult mice, Prp8 is expressed strongly in the testis and moderately in the ovary. in situ hybridization analysis revealed that Prp8 is preferentially expressed in the outer cell layer in the testis, probably in the spermatogonia and primary spermatocytes, and in granulosa cells in the ovary. In Caenorhabditis elegans, microinjection of a double stranded RNA corresponding to a portion of the Prp8 sequence results in the arrest of embryogenesis at the late-gastrulation stage. These results suggest that Prp8 plays an important role in reproduction and development. Prp8 was shown to bind to midkine (MK), a heparin-binding growth factor. Since Prp8 expression partially overlaps with the sites of action of MK, it is possible that binding to Prp8 is involved in part of MK signaling.

Aging↗

A serine/threonine kinase which causes apoptosis-like cell death interacts with a calcineurin B-like protein capable of binding Na(+)/H(+) exchanger.

We surveyed proteins capable of binding to the cytoplasmic domain of Na(+)/H(+) exchanger (NHE)1 in a rat brain cDNA library with the yeast two-hybrid system. One clone obtained coded for a protein reported previously as a human calcineurin homologous protein (CHP). Since CHP is homologous to the regulatory subunit B of calcineurin, we expected a possible interacting partner of CHP like the catalytic subunit of calcineurin (calcineurin A), and surveyed this putative partner again with the yeast two-hybrid system. A clone thus obtained coded for a kinase, which is basically the same as that reported for human DRAK2. Overexpression of the rat homologue of DRAK2 caused apoptosis-like cell death of NIH3T3 cells, which was dependent on the kinase activity, confirming the previous result for DRAK2. The purified CHP and rat DRAK2 proteins synthesized in Escherichia coli could bind in vitro. CHP and rat DRAK2 expressed in COS-7 cells were found to be localized in the Golgi apparatus and nucleus, respectively. Some of them was also found in the membrane peripheral region. When they were co-expressed in the same cells, most of CHP moved to the nucleus where rat DRAK2 is located, suggesting in vivo interaction of these proteins. However, minor but significant fractions of both proteins were also found in the membrane peripheral region. Rat DRAK2 is expressed highly in thymus, spleen, and testis, where the apoptosis plays an important role in physiology.

Amino Acid Sequence↗

The effects of danaparoid, dalteparin and heparin on tissue factor-induced experimental disseminated intravascular coagulation and bleeding time in the rat.

Danaparoid and heparin, on the basis of anti-activated factor X (anti-FXa) activity, were equipotent in accelerating the rate of interaction of FXa and antithrombin III. In rat tissue factor-induced disseminated intravascular coagulation (DIC) models, an intravenous administration of danaparoid inhibited the decrease in plasma fibrinogen and platelet counts and the increase in serum fibrinogen degradation products. Expressed on the basis of anti-FXa activity, these effects were comparable with those of dalteparin and heparin. In rat mesenteric small artery and vein, less bleeding was observed after intravenous administration of danaparoid than after dalteparin or heparin. Danaparoid did not affect adenosine diphosphate- or collagen-induced platelet aggregation, and showed weaker inhibitory effects on aggregation induced by thrombin, or collagen + thrombin, than did dalteparin or heparin. These findings suggest that danaparoid may be useful for the prevention of DIC and has less tendency to cause bleeding than dalteparin or heparin, probably as a result of its weaker ability to inhibit platelet aggregation.

Animals↗

Optical coherence tomography for the noninvasive evaluation of the cornea.

PURPOSE: To determine the optical coherence tomographic (OCT) characteristics of normal corneas and to characterize the OCT images of abnormal corneal lesions. METHODS: Eleven eyes from 10 patients were examined at the Cornea Service of the Nagoya University Hospital: 4 had corneal pathologies, 4 underwent keratoplasty, and 2 were normal controls; 1 enucleated eye was also examined. OCT (OCT 2000 Zeiss-Humphrey) was used to study the normal cornea and various corneal abnormalities. We compared the OCT images to the observations made by slit-lamp biomicroscopy. RESULTS: Fluid spaces were detected as black images. A highly reflective reflex was observed at the interface of different tissues, and intensive backscattering (reflex) was seen when the incident ray hit the laminated layers vertically. Corneal opacities were not clearly imaged when they were diffuse and mild, or when they were arranged axially in a small area, as was the scar of the graft-host junction. It was possible to obtain images from the region of the cornea that was not clearly visible by slit-lamp examination because of a corneal opacity. CONCLUSION: OCT is a noncontact and noninvasive technique that can be performed safely on diseased corneas. OCT can provide objective documentation of corneal disorders that cannot be obtained by slit-lamp examination. The use of OCT in conjunction with other conventional instruments should provide a more complete image of the cornea.

Adult↗

Surface ultrastructure of collagen fibrils and their association with proteoglycans in human cornea and sclera by atomic force microscopy and energy-filtering transmission electron microscopy.

PURPOSE: We aimed to investigate the possible association of proteoglycans with D-periodic collagen fibrils in the human cornea and sclera, using energy-filtering transmission electron microscopy (EF-TEM) and atomic force microscopy (AFM). METHODS: Human cornea and sclera were digested with keratanase to eliminate keratan sulfate proteoglycans (KSPGs). For EF-TEM observation, surface proteoglycans were detected by cupromeronic blue (CB) staining. For AFM observation, cornea and sclera were treated with sodium hydroxide before and after keratanase digestion, and the surface topology of collagen fibrils was analyzed. RESULTS: With CB staining, numerous CB-positive short filaments of surface proteoglycans (proteoglycan filaments) were observed in the interfibrillar spaces of cornea and sclera associated with collagen fibrils. AFM imaging showed that the depth and periodicity of D-periodic collagen fibrils in keratanase-treated corneal collagens were deeper and more regular than in untreated ones. Moreover, the depth and periodicity of keratanase-untreated corneal collagens were shallow and irregular in comparison with keratanase-untreated scleral collagens. On the other hand, there was no difference in depth or regularity between keratanase-treated and -untreated scleral collagen fibrils. Using AFM imaging, additional thin grooves sub-bands were detected on the surface of keratanase-treated corneal collagen fibrils. The grooves were not detected in keratanase-untreated collagen fibrils nor in scleral collagen fibrils with or without keratanase digestion. Comparing densitometry waves, the grooves of D-periodic corneal collagen sub-bands corresponded to a and c bands. CONCLUSION: Using AFM and EF-TEM to study corneal and scleral collagen fibrils and their association with proteoglycans, we conclude that KSPG is found in ample amounts in the human cornea in comparison with sclera. Moreover, we topologically detected KSPG attached to a and c bands of collagen fibrils.

Aged↗

Suppression of gingival inflammation induced by Porphyromonas gingivalis in rats by leupeptin.

In this study, we developed a procedure to produce gingivitis in rats by inoculation of Porphyromonas gingivalis and studied the contribution of the bacterial cysteine proteinases, Arg-gingipain (Rgp) and Lys-gingipain (Kgp), to the pathology in the gingiva. To adhere the bacterium to periodontal tissues, a cotton thread was inserted between the first and second molar of right maxillary sites of rats. Rats in group A were administered with vehicle alone after bacterial (strain W83) inoculation. In group B, the bacteria were inoculated in combination with leupeptin, a potent inhibitor of Rgp and Kgp, and then leupeptin alone was administered the week after. Rats in group C were administered leupeptin for 6 weeks after bacteria inoculation. All left maxillary gingiva in three groups showed no inflammatory changes. Right maxillary gingiva of group A showed most of the clinical landmarks of gingivitis. Leupeptin exhibited only a little inhibitory effect on this gingivitis in group B, whereas it had a strong inhibitory effect on the inflammation in group C. These results suggest that P. gingivalis-induced gingivitis is attributable to Rgp and Kgp and that leupeptin is more effective in the late phase than the early stage of gingivitis.

Adhesins, Bacterial↗

Revised sequence and expression of cyclin B cDNA from the starfish Asterina pectinifera.

Cyclin B cDNA was cloned from the ovary of the starfish Asterina pectinifera and analyzed by RT-PCR and 3'- and 5'-RACE techniques. The cDNA consists of a 0.13-kb upstream untranslated region, a 1.22-kb coding region, and a 0.86-kb downstream untranslated region. The open reading frame encoded a polypeptide of 404 amino acid residues with a calculated molecular weight of 45,692. All the characteristic sequences, such as destruction and cyclin boxes, cyclin B motif, and cytoplasmic retention and nuclear export signals, were found in the newly cloned cyclin B cDNA. The deduced amino acid sequence of the cyclin B cDNA was highly homologous in the middle and carboxy terminal regions to that from mature eggs of the same organism, but quite different in the amino terminal region. Evidence was obtained which suggested that this cyclin B is expressed in immature and maturing oocytes and is the same as that cloned from mature eggs.

Amino Acid Sequence↗

Hypertension and impairment of endothelium-dependent relaxation of arteries from spontaneously hypertensive and L-NAME-treated Wistar rats.

Effects of chronic treatment of normotensive Wistar rats with N(omega)-nitro-L-arginine methyl ester (L-NAME) on blood pressure and on endothelium-dependent relaxation of the aorta, carotid and iliac arteries were studied. The endothelium-dependent relaxation was compared in arteries from normotensive Wistar Kyoto rats (WKY) and genetically hypertensive rats (stroke-prone spontaneously hypertensive rats, SHRSP). Chronic treatment of normotensive Wistar rats with L-NAME caused an elevation of blood pressure. The elevated blood pressure at 15 weeks of age was significantly higher in these animals than that of untreated Wistar rats, but lower than that of SHRSP. Endothelium-dependent relaxation of the arteries induced by acetylcholine (ACh) was almost abolished by chronic treatment with L-NAME. The remaining small relaxation in arteries from L-NAME-treated rats was completely inhibited by application of L-NAME (10(-4) M). In such preparations, higher concentrations of ACh induced a contraction, which was abolished by removal of the endothelium or by an application of indomethacin (10(-5) M). Endothelium-independent relaxation induced by sodium nitroprusside was similar between preparations from untreated and L-NAME-treated Wistar rats. Endothelium-dependent relaxation was significantly impaired in preparations from SHRSP, when compared with that in those from WKY. However, the impairment was less prominent in preparations from SHRSP than in those from L-NAME-treated rats. These results suggest that the impairment of endothelium-dependent relaxation in the arteries from L-NAME-treated rats is not due to the elevated blood pressure resulting from the chronic treatment, and that impairment of NO synthesis by the endothelium does not play a major role in the initiation of hypertension in SHRSP.

Animals↗

Difference in effects of stretch on depressive effect of endothelium-derived nitric oxide on noradrenaline- and high-K+-induced contractions between the aortae from normotensive and spontaneously hypertensive rats.

Difference in effects of stretch tension on endothelium-derived nitric oxide (EDNO)-dependent depression of noradrenaline (NA)- and high-K+-induced contraction between the aortae from normotensive Wistar Kyoto rats (WKY) a nd stroke-pronespontaneously hypertensive rats (SHRSP) was studied. NA-induced contraction in preparations both from WKY and SHRSP was augmented in the presence of N(omega)-nitro-L-arginine (L-NNA). This augmentation was minimized when the spontaneous tone, which was more prominent in preparations from SHRSP, was subtracted and the effects of L-NNA became less prominent in preparations from SHRSP. The effects of L-NNA were maximal at the stretch tension of 15 mN and, then, decreased as stretch tension increased in both preparations when the spontaneous tone was subtracted. The effects of L-NNA were less prominent when the contraction was initiated by high-K+, although the effects of stretch on high-K+-induced contraction were similar to that of NA-induced contraction. These results suggested 1) that both NA- and high-K+-induced contractions are depressed by EDNO, 2) that the release of EDNO induced by high-K+ is less than that by NA, 3) that increase in stretch tension decreases the release of EDNO, and 4) that the depressive effect of EDNO on contraction is impaired in the aorta of SHRSP.

Animals↗

Therapeutic effects of azathioprine in combination with low-dose prednisolone in patients with intractable autoimmune hepatitis type 1.

This study evaluated the effects of azathioprine in combination with low-dose prednisolone in the management of patients with intractable autoimmune hepatitis. Thirteen patients with intractable autoimmune hepatitis who had an incomplete or arrested response to conventional prednisolone therapy, or who relapsed during prednisolone maintenance therapy were additionally administered 50 or 100 mg/day of azathioprine in combination with prednisolone. This regimen reliably induced complete remission in 12 of 13 patients, and these 12 remained in remission during the follow-up period with maintenance therapy of 50 mg/day of azathioprine in combination with 5 mg/day of prednisolone. The findings of the current study indicate that the azathioprine and low-dose prednisolone combined therapy may offer a satisfactory alternative therapy for patients with intractable autoimmune hepatitis who have an incomplete or arrested response to conventional prednisolone therapy, or who relapse during prednisolone maintenance therapy.

Adult↗