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Biomedical subjects

S Tsukahara

Publications and source records attributed to S Tsukahara.

At least 19 recordsLinked to original sources
PubMed2000

Optical coherence tomography of orange-red subretinal lesions in eyes with idiopathic polypoidal choroidal vasculopathy.

PURPOSE: To study the cross-section images of orange-red lesions in eyes with idiopathic polypoidal choroidal vasculopathy and compare their protrusions quantitatively with those of serous retinal pigment epithelium detachment. METHODS: Optical coherent tomography images scanning remarkable orange-red lesions in eyes with idiopathic polypoidal choroidal vasculopathy and serous retinal pigment epithelium detachment in age-related macular degeneration or central serous chorioretinopathy were prospectively recorded. The correlation between the base diameter and the height of the lesions was analyzed. RESULTS: Optical coherence tomography images show prominent anterior protrusion of the orange-red lesions. The ratio of the height to the base diameter of the orange-red lesions in eyes with idiopathic polypoidal choroidal vasculopathy is 0.32+/-0.05, and it is significantly larger than lesions in eyes with serous retinal pigment epithelium detachment (0.18+/-0.05, P< .001). CONCLUSION: The orange-red lesions in eyes with idiopathic polypoidal choroidal vasculopathy have a more sharply peaked shape than serous retinal pigment epithelium detachment as a subretinal structure, suggesting polypoidal vascular lesions in eyes with idiopathic polypoidal choroidal vasculopathy are situated beneath the Bruch membrane and covered anteriorly with both the retinal pigment epithelium and the Bruch membrane.

Aged↗
PubMed2000

Localization of glucokinase-like immunoreactivity in the rat lower brain stem: for possible location of brain glucose-sensing mechanisms.

Pancreatic glucokinase (GK) is considered an important element of the glucose-sensing unit in pancreatic beta-cells. It is possible that the brain uses similar glucose-sensing units, and we employed GK immunohistochemistry and confocal microscopy to examine the anatomical distribution of GK-like immunoreactivities in the rat brain. We found strong GK-like immunoreactivities in the ependymocytes, endothelial cells, and many serotonergic neurons. In the ependymocytes, the GK-like immunoreactivity was located in the cytoplasmic area, but not in the nucleus. The GK-positive ependymocytes were found to have glucose transporter-2 (GLUT2)-like immunoreactivities on the cilia. In addition, the ependymocytes had GLUT1-like immunoreactivity on the cilia and GLUT4-like immunoreactivity densely in the cytoplasmic area and slightly in the plasma membrane. In serotonergic neurons, GK-like immunoreactivity was found in the cytoplasm and their processes. The present results raise the possibility that these GK-like immunopositive cells comprise a part of a vast glucose-sensing mechanism in the brain.

Animals↗
PubMed2000

Anti-human immunodeficiency virus type 1 of a two-strand-system targeted to the polypurine tract.

The polypurine tract (PPT) is highly conserved among the known human immunodeficiency virus (HIV)-1 strains, and is a possible target for triplex formation. We show the effects of triple-helix formation by assays of primer extension inhibition in vitro, using a two-strand-system (FTFOs) targeted to the PPT of HIV-1. The two-stranded composition of a triple-helix is thermodynamically and kinetically superior to the three-strand-system. The FTFOs inhibited the RT activity in a sequence-specific manner, i.e., the triplex actually formed at the PPT and blocked the RT. The FTFOs containing the phosphorothioate groups at the antisense sequences showed greater 3'-exonuclease resistance. In the observation of the FITC-DsDGloopT5-37 with MOLT-4 cells by a confocal laser scanning microscope, diffuse fluorescence was apparently observed in the cytoplasm and nucleus. However, weak fluorescence was observed within the cytoplasm and nucleus of MOLT-4 cells treated with the antisense phosphorothioate oligonucleotides (S-ODN-gag-AUG). In HIV-1 infected MOLT-4 cells, the FTFOs containing the phosphorothioate groups at the antisense sequence sites and guanosine rich parts within the third Hoogsteen base pairing sequence inhibit the replication of HIV-1 more effectively than the antisense oligonucleotides, indicating sequence-specific inhibition of HIV-1 replication.

Amino Acid Sequence↗
PubMed1999

Prevention of inhibitory effect of dorsal raphe nucleus lesions on ovulation and LH surge by 5-HT 2A/2C receptor agonists in female rats.

In order to investigate the role of the dorsal raphe nucleus and the serotonergic system in the regulation of ovulation, the number of ova and plasma luteinizing hormone (LH) concentrations were measured in female rats after making lesions in this nucleus (DRL) and/or treatment with 5-hydroxytryptamine (5-HT) receptor agonists or antagonists. DRL or sham lesion was made on the afternoon of proestrous (12:00-14:00 h) under ether anesthesia and the number of ova in the oviduct was counted on the next estrous and diestrous morning. In some animals, blood samples were taken via the atrial cannula during the proestrous evening for the radioimmunoassay of LH. All intact control and sham-operated females ovulated and plasma LH increased between 19:00 and 21:00 h. In contrast, ovulation was seen in only 36% of DRL rats. LH surge did not occur in this group. However, 80% of DRL rats ovulated after treatment with (+/-)-1-(4-iodo-2,5-dimethoxyphenyl)-2-aminopropane hydrochloride [(+/-)-DOI; a 5-HT 2A/2C receptor agonist] at 15:00 h on proestrous day. LH surge was also observed in the DRL rats with (+/-)-DOI. On the other hand, only 8% of DRL rats ovulated after treatment with buspirone (5-HT 1A receptor agonist). Furthermore, when mianserin (5-HT 2A/2C receptor antagonist) was administered at 16:00 h on proestrous day, ovulation was not seen in all rats without DRL. These results suggest that the dorsal raphe nucleus plays an important role in induction of LH surge and ovulation and the 5-HT 2A/2C receptor system is involved in this mechanism.

Animals↗
PubMed1999

Inhibition of HIV-1 replication by a two-strand system (FTFOs) targeted to the polypurine tract.

Reverse transcription of HIV-1 vRNA into the double-stranded DNA provirus involves initiation of plus-strand DNA synthesis at the polypurine tract (PPT) by reverse transcriptase (RT). The PPT is highly conserved among the known human immunodeficiency virus (HIV-1) strains and is a possible target for triplex formation. We show the effects of triple-helix formation by assays of primer extension inhibition in vitro, using a two-strand system (foldback triplex-forming oligonucleotides (FTFOs)) targeted to the PPT of HIV-1. The two-stranded composition of a triple-helix is thermodynamically and kinetically superior to the three-strand system. The FTFOs inhibited the RT activity in a sequence-specific manner, i.e. the triplex actually formed at the PPT and blocked the RT. The FTFOs containing the phosphorothioate groups at the antisense sequences showed greater 3'-exonuclease resistance. In HIV-1-infected MOLT-4 cells, the FTFOs containing the phosphorothioate groups at the antisense sequence sites and guanosine rich parts within the third Hoogsteen base-pairing sequence inhibit the replication of HIV-1 more effectively than the antisense oligonucleotides, indicating sequence-specific inhibition of HIV-1 replication.

Anti-HIV Agents↗
PubMed1999

Inhibition of restriction endonuclease cleavage by triple helix formation with RNA and 2'-O-methyl RNA oligonucleotides containing 8-oxo-adenosine in place of cytidine.

The ability of homopyrimidine oligoribonucleotides (RNA) and oligo-2'-O-methyl-ribonucleotides (2'-O-methyl RNA) containing 8-oxo-adenosine (AOH) and 8-oxo-2'-O-methyl (AmOH) adenosine to form stable, triple-helical structures with sequences containing the recognition site for the class II-S restriction enzyme, Ksp632-I, was studied as a function of pH. The AOH- and AmOH-substituted RNA and 2'-O-methyl RNA oligonucleotides were shown to bind within the physiological pH range in a pH-independent fashion, without a compromise in specificity. The substitutions of three cytidine residues with AOH showed higher endonuclease inhibition than the substitution of either one or two cytidine residues with AOH. In particular, the 2'-O-methyl RNA oligonucleotide with only one cytidine substituted with AmOH showed higher endonuclease inhibition than the homopyrimidine RNA and 2'-O-methyl RNA oligonucleotides and the RNA oligonucleotides containing either one or two AOH moieties. Furthermore, the AmOH-substituted 2'-O-methyl RNA oligonucleotides were stable (53%) after an incubation in 10% fetal bovine serum for 8 h, whereas the RNA oligonucleotides were completely degraded. Increased resistance to nucleases is observed with the introduction of 2'-O-methylnucleosides. This stabilization should help us to design much more efficient third strand homopyrimidine oligomer and antisense nucleic acid-based antiviral therapies, which could be used as tools in cellular biology.

Adenosine↗
PubMed1999

Morphological characterization of relationship between gap junctions and gonadotropin releasing hormone nerve terminals in the rat median eminence.

The present study aimed to reveal possible morphological relationships between gonadotropin-releasing hormone (GnRH) nerve terminals and gap junctions in the median eminence. Coronal brain sections from castrated male rats were dual immunostained with GnRH and either connexin 26, 32, or 43, and examined by confocal laser microscopy. Connexin 43-immunoreactive puncta were distributed between GnRH-immunoreactive fibers, and some of them were colocalized with GnRH-immunoreactivities. Dual immunostaining with connexin 43 and glial fibrillary acidic protein revealed that most of the puncta were located in astrocytes. At the immunoelectron microscopic level, connexin 43-immunoreactivities were mainly located on the plasma membranes of glial-like processes. Few connexin 26- or connexin 32-immunoreactivities were found in the median eminence. The present results indicate the possibility that gap junctions play a role in the GnRH release at the median eminence.

Animals↗
PubMed1999

Progression of defects in the central 10-degree visual field of patients with retinitis pigmentosa and choroideremia.

PURPOSE: To determine the progression of defects in the central 10-degree visual field in patients with retinitis pigmentosa and choroideremia by means of univariate linear regression with respect to the mean deviation. METHODS: In a retrospective study, results of automated static perimetry in the central 10-degree visual field were reviewed for patients with retinitis pigmentosa and choroideremia. Univariate linear regression of mean deviation was undertaken for each eye that had field data from at least five tests in addition to the prior static perimetric experience during a period of 3.5 years or more. Data from 30 eyes of 16 patients fulfilled eligibility requirements; the mean number of fields was 5.8 (range, five to 10) and the mean follow-up period was 64 months (range, 42 to 97 months). Eyes were classified as progressive if the regression coefficient was negative and significantly different from zero, with a P value of less than .05. RESULTS: Among 14 patients in whom both eyes were eligible for univariate linear regression analysis, a statistically significant progression was demonstrated in both eyes in four patients, only in one eye in five patients, and in neither eye in five patients. One of two patients, in whom only one eye was eligible for the study, showed significant progression in the eye. Overall, 14 (47%) of 30 eyes showed statistically significant progression with respect to the mean deviation. No eye showed significant improvement. CONCLUSIONS: Automated static perimetry of the central 10-degree visual field measured approximately once or twice a year demonstrated the progressive nature of the disease with the use of univariate linear regression of mean deviation in 47% of eyes with retinitis pigmentosa and choroideremia during the follow-up period of 3.5 years or more. These results may be useful in understanding the clinical course of the diseases and counseling patients.

Adolescent↗
PubMed1999

Optical coherence tomography of idiopathic polypoidal choroidal vasculopathy.

PURPOSE: To document and study the cross-sectional structures of polypoidal elements using optical coherence tomography in eyes with idiopathic polypoidal choroidal vasculopathy. METHODS: Optical coherence tomography images of two eyes with idiopathic polypoidal choroidal vasculopathy were correlated with slit-lamp biomicroscopic findings, fundus photographs, fluorescein angiograms, and indocyanine green angiograms. RESULTS: Cross-sectional optical coherence tomographic images of retinochoroidal structures showed prominent anterior protrusion of the orange subretinal mass corresponding to the polypoidal structure in the indocyanine green angiogram. Hemorrhagic detachment of the retinal pigment epithelium was contiguous with the cone-shaped nodule beneath the retinal pigment epithelium in one eye, and an apparent discontinuity was observed in the highly reflective layer that delineates the polypoidal structure. CONCLUSIONS: Some of the polypoidal structures in eyes with idiopathic polypoidal choroidal vasculopathy are anteriorly protruding lesions in the inner choroid that may cause serosanguineous detachment of the retinal pigment epithelium through damage of the overlying Bruch membrane, retinal pigment epithelium, and the adhesion between them. Cross-sectional optical coherence tomographic images may increase understanding of the pathophysiology of idiopathic polypoidal choroidal vasculopathy.

Aged↗
PubMed1999

Optical coherence tomography of cystoid macular edema associated with retinitis pigmentosa.

PURPOSE: To detect cystoid macular edema in consecutive eyes with retinitis pigmentosa by means of optical coherence tomography and to study the correlation between cross-sectional structures and angiographic findings in cystoid macular edema. METHODS: In a prospective study, cross-sectional images through the fovea were evaluated by means of optical coherence tomography in 89 phakic eyes of 46 patients with retinitis pigmentosa. Eyes showing cystoid appearance in the macula in the optical coherence tomographic images were further studied with measurement of the dimensions of cystoid lesions and with a fluorescein angiogram either at 18 minutes after dye injection or later. RESULTS: Cystoid lesions were observed in the macula in optical coherence tomographic images in 12 eyes in six (13%) of 46 patients. In these eyes, the width of total area of the cystoid lesions was positively correlated with the grade of fluorescein angiogram (Spearman rank correlation coefficient, r = .629; P = .029), but the thickness of the neurosensory retina at the center of the fovea was not. Among three variables for grading cystoid macular edema, consisting of angiographic grade, thickness of the neurosensory retina at the center of the fovea, and width of total area of the cystoid lesions, only the last measure was significantly correlated with best-corrected visual acuity (Pearson correlation coefficient, r = .693; P = .012). CONCLUSION: Cystoid macular edema in eyes with retinitis pigmentosa could easily be detected with the use of optical coherence tomography independent of the angiographic degree of dye leakage. The size of cystoid lesions demonstrated in the optical coherence tomographic images, especially the thickness of the neurosensory retina at the center of the fovea, was not necessarily correlated with the angiographic grading of dye leakage. Measurement of the width of total area of the cystoid lesions in the optical coherence tomographic images is significantly correlated with the loss of visual acuity.

Adolescent↗
PubMed1999

Optical coherence tomography of successfully repaired idiopathic macular holes.

PURPOSE: To present the cross-sectional retinal imaging results of optical coherence tomography in eyes with successfully repaired idiopathic macular hole and their relevance to visual recovery. METHODS: We studied 33 eyes with successful repair of an idiopathic macular hole through vitrectomy and fluid-gas exchange from 32 patients (11 men and 21 women) with ages ranging from 48 to 78 years, with a median age of 66 years. Preoperative conditions in eyes with primary surgery disclosed nine eyes with stage 2, 14 eyes with stage 3, and four eyes with stage 4 macular hole. An additional six eyes underwent a second surgery because the previous surgery was unsuccessful. Measurement of best-corrected visual acuity, slit-lamp biomicroscopy with fundus contact lens, fundus photographs, and optical coherence tomographic examination were performed between 6 and 9 months after surgery in 29 eyes and between 15 and 36 months after surgery in four eyes. RESULTS: Optical coherence tomographic images of the repaired macular holes were categorized into three patterns. U-type (normal foveal contour; 13 eyes) showed mildly to moderately backscattering layers with a smooth circular surface covering retinal pigment epithelium and choriocapillaris layers. In eyes with V-type (steep foveal contour; 13 eyes), the retinal pigment epithelium and choriocapillaris layers were covered with moderately backscattering layers with a notch. W-type (foveal defect of neurosensory retina; seven eyes) showed abruptly or gradually terminating sensory retinal layers to expose the surface of the retinal pigment epithelium and choriocapillaris layers. Postoperative acuity was well correlated with these patterns of optical coherence tomographic images. CONCLUSION: Assessment of successfully repaired idiopathic macular holes with optical coherence tomographic images provides a useful correlation with postoperative visual recovery.

Aged↗
PubMed1999

Assessment of potential macular function using a color saturation discrimination test in eyes with cataract.

PURPOSE: To study the efficacy of a color saturation discrimination test (CSDT) in assessing potential macular function in eyes with cataract. SETTING: Yamanashi Medical University and Nirasaki City Hospital, Yamanashi, Japan. METHODS: In this prospective study of 200 consecutive eyes with cataract, the thresholds of color saturation discrimination on 6 hues were measured with the CSDT. The CSDT program was run on a personal computer and cathode-ray tube color monitor. The preoperative CSDT scores and postoperative visual acuities were compared. In addition, preoperative laser interferometry (LI) and postoperative CSDT were performed on selected patients and the results compared with their preoperative CSDT results. RESULTS: Patients who had a CSDT score of 11 or more had a significantly lower postoperative visual acuity (P < .0001). The prevalence of neuroretinal abnormality postoperatively increased with the preoperative CSDT score. The CSDT score was not correlated with preoperative visual acuity (P = 1409). Cataract severity had little influence on the CSDT score. The preoperative results of the CSDT and Ll were correlated with the postoperative acuity, with the correlation coefficient being stronger with the CSDT (r = -0.338 and 0.276 for CSDT and Ll, respectively). CONCLUSION: The CSDT was effective in evaluating neuroretinal pathology and predicting postoperative acuity.

Cataract↗
PubMed1999

Effect of corticotropin-releasing hormone antagonist on oestrogen-dependent glucoprivic suppression of luteinizing hormone secretion in female rats.

Pharmacological reduction of glucose availability with 2-deoxyglucose (2DG) suppresses pulsatile luteinizing hormone (LH) secretion in rats and growth-retarded lambs. Gonadal steroids enhance the glucoprivic suppression of LH secretion in rats. The present study determined if corticotropin-releasing hormone (CRH) plays a role in mediating oestrogen-dependent and -independent glucoprivic suppression of LH secretion. The study was conducted in ovariectomized (OVX) rats some of which received Silastic implants containing oestradiol-17beta (OE2) dissolved in peanut oil at 20 microg/ml to produce a physiological plasma level of OE2 (30 pg/ml). Seven days after ovariectomy, the rats were stereotaxically implanted with a guide cannula into the third cerebral ventricle. Seven days later, blood samples were collected through an indwelling atrial cannula every 6 min for 3 h for LH pulse determination. After the first hour of blood sampling, a CRH antagonist, [D-Phe12, Nle21,38]hCRF-(21-41), or vehicle was injected into the third cerebral ventricle through the implanted cannula before 2DG administration through the indwelling atrial cannula. Pulsatile LH secretion was suppressed by 2DG (200 mg/kg b.w.) in the vehicle-treated rats bearing OE2 implants. The CRH antagonist (5.65 nmol) blocked the suppressive effect of 2DG on pulsatile LH secretion in the OE2-treated OVX animals. On the other hand, in the absence of oestrogen, the effect of a twice greater dose of 2DG (400 mg/kg b.w.) was not blocked by five times greater amount of CRH antagonist (28.3 nmol). These results suggest the mechanisms mediating glucoprivic suppression of LH secretion involve two components: one is oestrogen-dependent and the other oestrogen-independent. CRH may be involved in the oestrogen-dependent component of glucoprivic suppression of LH secretion but not the oestrogen-independent one.

Animals↗
PubMed1999

Three-dimensional arrangement of collagen fibrils in human ciliary body.

The purpose of this study is to visualize the three-dimensional arrangement of collagen fibrils in aged human ciliary body and discuss their significance. The ciliary bodies obtained from two human eyes were treated with a NaOH cell-maceration method for 7 days, then prepared conventionally for light and scanning electron microscopy. The general morphology of the collagen tissue in the ciliary body appeared almost the same as that normally observed. Cellular elements were completely removed, but collagen fibrils were well preserved. In the stroma of the ciliary body, collagen fibrils were arranged irregularly. In the areas of the radial and circular ciliary muscles, considerable numbers of collagen fiber bundles were observed running in a circular direction. A honeycomb structure was seen in the pars plana, the walls and base of which were formed by interweaving collagen fibrils. The results suggested that collagen fibrils in the aged human ciliary body may be largely involved in the presbyopia.

Aged↗
PubMed1999

Metabolites of isopropyl unoprostone as potential ophthalmic solutions to reduce intraocular pressure in pigmented rabbits.

The intraocular metabolism of isopropyl unoprostone, a novel prostaglandin-related anti-glaucoma compound, was investigated using pigmented rabbits to clarify which metabolites are involved in actions in the eye. Tritium-labeled isopropyl unoprostone eyedrops were administered. The cornea, aqueous humor, iris, ciliary body and retina were then collected at 5, 15 or 30 min or at 2, 6 or 12 h after instillation. Isopropyl unoprostone and its metabolites were fractionated using high-performance liquid chromatography, and the radioactivity of each fraction was measured. Unmetabolized isopropyl unoprostone was never detected in any sample at any time point. In the cornea, only the de-esterificated metabolite, M1, and the further metabolized compound, M2, were detected; and the concentrations of these metabolites decreased with time. In the aqueous humor, M1, M2 and another metabolite, M3, were detected, with peak concentrations of M1 at 30 min and M2 at 2 h. The iris and ciliary body showed a similar metabolism with peak concentrations of M1 and M2 at 30 min. In the aqueous humor, iris and ciliary body, M2 was the dominant metabolite from 30 min. In the retina, only total radioactivity was detected. These results indicate that the main metabolites involved in actions in the eye are M1 and M2.

Animals↗
PubMed1999

Measurement of adenylate cyclase activity in the minute bovine ciliary epithelial cells during the pharmacological stimulation of adrenergic and cholinergic receptors.

Although essential to the secretion of aqueous humor, little is known about the signal transduction underlying postreceptor adrenergic and cholinergic processes in the ciliary epithelium. We adopted a highly sensitive fluorometric assay technique in order to examine adenylate cyclase activity in minute membrane preparations made from the bovine ciliary epithelial cells. The protein concentration of the preparation was 3-5 mg/ ml. Norepinephrine (10(-7), 10(-6) and 10(-5) M) and carbachol (10(-7) and 10(-5) M) were incubated with 10 microl of membrane preparation to analyze the extent of the receptor-coupled influences on the adenylate cyclase activity. Meanwhile, forskolin (10(-5) M) was used to estimate the maximum adenylate cyclase activity. After the initial enzymatic destruction of noncyclic adenine nucleotides and phosphorylated metabolites, the diester linkage of cyclic AMP was cleaved and then converted to ATP. The ATP was enzymatically amplified to about 10,000 times of fructose-6-phosphate. The NADPH, formed when the fructose-6-phosphate was converted to 6-phosphogluconolactone, was measured fluorometrically. Basal and forskolin-stimulated maximum adenylate cyclase activities (pmol/mg protein/min) were 29.6+/-7.6 and 86.6+/-7.2 (mean+/-SE), respectively. Norepinephrine increased the adenylate cyclase activity in a dose-dependent manner, while carbachol hardly affected the activity. These results indicate that the adenylate cyclase activity can be measured in the minute ciliary epithelial cells and, moreover, that the current assay can be applied to assess the efficacy of newly available ophthalmic solutions or systemic drugs influencing adenylate cyclase activity in a discrete portion in the eye.

Adenylyl Cyclases↗
PubMed1998

Inhibitory effect of neural transections of dorsal raphe nucleus on induction of nocturnal prolactin surge by vaginal stimulation in ovariectomized rats.

The effect of complete (CC), anterior (AC) or posterior (PC) cut of the dorsal raphe nucleus (DRn) on induction of the nocturnal prolactin (PRL) surge by electrical vaginal stimulation (VS) was investigated in ovariectomized rats. Plasma level of PRL was measured by radioimmunoassay before and after VS. The data revealed that PRL levels increased in early morning on the day following VS in the rats without brain surgery or with sham-operation. In contrast, the nocturnal PRL surge did not occur in the CC, AC, or PC rats. These results suggest that both the anterior and the posterior fibers of the DRn plays an important role in induction of nocturnal PRL surge by VS in ovariectomized rats.

Analysis of Variance↗