Biomedical subjects

H Ohkuma

Publications and source records attributed to H Ohkuma.

At least 37 records · Page 2Linked to original sources

PubMed1999

Effects of intracisternal methylprednisolone on lipid peroxidation in experimental subarachnoid haemorrhage.

The efficacy of intracisternal methylprednisolone was examined on lipid peroxidation in a canine subarachnoid haemorrhage (SAH) model. The concentration of lipid peroxides increased significantly in the cerebrospinal fluid (CSF) supernatant on Day 4, and also in the arterial wall on Day 7. Intracisternal administration of methylprednisolone reduced markedly the products of lipid peroxidation both in CSF and in the arterial wall. The findings suggest that lipid peroxidation might play a significant role in the genesis of vasospasm after SAH, and that direct administration of methylprednisolone into the subarachnoid space might reduce lipid peroxides in the arterial wall and so influence the prevention of vasospasm.

Animals↗

PubMed1999

Antisense preproendothelin-oligoDNA therapy for vasospasm in a canine model of subarachnoid hemorrhage.

OBJECT: The purpose of this study is twofold: 1) to test antisense genetic techniques used in the prevention of cerebral vasospasm in a canine model of subarachnoid hemorrhage (SAH), targeting the endothelin-1 (ET-1) gene; and 2) to determine if fibrinolysis of subarachnoid clot with recombinant tissue plasminogen activator (rtPA) could enhance the effect of antisense treatment. METHODS: A total of 39 dogs were studied in this experiment. Placebo (six animals), rtPA (six animals), antisense preproET-1 oligodeoxynucleotide (ASOD; five animals), or rtPA plus ASOD (combined treatment; six animals) was injected into the cisterna magna 30 minutes after a second SAH was induced on the 2nd day of the experiment. The animals were observed until Day 7, when they underwent follow-up angiography and then were killed; their basilar arteries were removed for analysis. Control animals included in this study (two animals in each group) received placebo, rtPA, ASOD, or rtPA plus ASOD without induction of SAH, or rtPA with mismatched (nonsense) preproET-1 oligodeoxynucleotide following SAH. Six additional dogs were analyzed earlier following SAH. Dogs that received placebo developed severe vasospasm (51+/-8% of baseline caliber). Administration of ASOD alone resulted in a mild reduction in vasospasm (64+/-13% of baseline caliber) and rtPA alone resulted in a moderate reduction in vasospasm (81+/-5% of baseline caliber); however, the combined therapy of rtPA plus ASOD almost completely prevented vasospasm (95+/-6%, of baseline caliber), which was significantly different from all other groups (p < 0.05). Morphological analysis of the basilar arteries yielded results similar to angiography with respect to vasospasm severity. The ASOD treatment combined with rtPA resulted in reduced ET-1 expression, as demonstrated by immunohistochemical staining of the arteries, and reduced preproET-1 levels on Day 4, as measured by reverse transcription-polymerase chain reaction. Nonsense DNA sequences had no effect on the vessels. CONCLUSIONS: Antisense preproET-1 oligodeoxynucleotide treatment, when combined with clot lysis caused by rtPA, reduced vasospasm in the canine model of SAH, and this effect appeared to be related to reduced ET-1 synthesis. The results of this experiment support a causative role for ET-1 early in the course of vasospasm development in dogs. The apparent additive therapeutic effects of antisense and fibrinolytic treatments could be due to clot lysis, which allows better delivery of oligodeoxynucleotides to arteries within the subarachnoid space.

Animals↗

PubMed1999

[Intraoperative electrophysiological monitoring for functional preservation of the cauda equina during lumbosacral surgery].

Adequate electrophysiological techniques to monitor function of the cauda equina have been proposed for surgery in patients with lumbosacral lipoma or myeloschisis. Motor fibers were identified by electrical stimulation in the operating field with bipolar rectangular impulses of 200 mu sec duration at 2 Hz under 5 mA and compound muscle action potentials (CMAPs) recorded from the leg and anal muscles. By recording CMAPs from the tibialis anterior, the biceps femoris, the gastrocnemius, and the external anal sphincter muscles, all of the roots from the fourth lumbar to the fourth sacral segment were continuously monitored. To spare recording channels, recordings were obtained from the right versus the left side. In our institute, 5 patients have undergone lumbosacral surgery while using this monitoring system, and the results indicated that there was no postoperative neurological exacerbation in any of the cases. According to a combination of the CMAPs produced by stimulation, the segment of the stimulated motor root could be identified electrophysiologically. Monitoring of somatosensory evoked potentials was not performed because this would have required too much time and would have prolonged surgery. However, some sensory fibers, which appeared to be posterior roots of the cauda equina on intraoperative inspection, could be identified indirectly with CMAPs recording because of current spreading from the stimulation to motor fibers.

Action Potentials↗

PubMed1998

Novel mechanism by which hemoglobin induces constriction of cerebral arteries.

Since oxyhemoglobin (OxyHb) is implicated in the pathogenesis of cerebral vasospasm, we have investigated the role of protein tyrosine phosphorylation in OxyHb-mediated signalling in canine cerebral arteries and cultured canine cerebrovascular smooth muscle cells. OxyHb produced a contraction of basilar artery preparations, which was reversed by genistein, an inhibitor of tyrosine kinases, and PD098059, an inhibitor of mitogen-activated protein kinase. In cerebrovascular smooth muscle cells, OxyHb induced tyrosine phosphorylation of 42, 46, 54-60 and 80-100 kDa proteins with a time-course which paralleled the contractile action of OxyHb, suggesting that these events might be functionally linked. The 42 and 60 kDa proteins were immunologically related to the mitogen-activated protein kinase, extracellular signal regulated protein kinase (ERK2), and to p60c-Src (c-Src), respectively. The increase in protein tyrosine phosphorylation was attenuated by genistein, and the phosphorylation of the 42 kDa protein (ERK2) was inhibited by PD098059. These results suggest that OxyHb-mediated signalling utilizes a protein tyrosine kinase-based mechanism.

Animals↗

PubMed1998

Proposal of a high-field superconducting wiggler for a slow positron source at SPring-8.

A low-energy positron beam is a unique probe of materials. In high-energy electron and positron storage rings it is possible to generate intense synchrotron radiation with a photon energy of 1-3 MeV by installing a high-field (8-10 T) superconducting wiggler. High-energy photons are converted to low-energy positrons by using a suitable target-moderator system. For an 8 GeV electron storage ring at a beam current of 100 mA, final yields are estimated to be about 10(8)-10(10) slow-e(+) s(-1) or larger depending on the moderation efficiency, with the size of the positron source 10(1)-10(2) cm(2). In the present work a wiggler magnetic system of 10 T is proposed. The main parameters of the superconducting wiggler are presented.

Journal Article↗

PubMed1998

Apoptosis of photoreceptor cells in ornithine-induced retinopathy.

BACKGROUND: The intravitreal injection of ornithine produces selective damage to the retinal pigment epithelium (RPE) and results in a loss of RPE, choriocapillaris and photoreceptor cells. To elucidate the mechanism of secondary retinal atrophy, we investigated the presence of apoptotic cells in a rat model of ornithine-induced retinopathy. METHODS: At 6 and 12 h and 1, 2, 4, 7, 14 and 28 days after an intravitreal injection of L-ornithine hydrochloride in rat eyes, we removed the eyes and subjected them to histopathological examination. We detected apoptotic cells by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate digoxigenin nick end labeling (TUNEL) assay, which stains the 3'-OH ends of fragmented DNA. We used electron microscopy to detect the apoptotic cells morphologically. RESULTS: RPE cells were selectively damaged immediately after ornithine administration. TUNEL-positive photoreceptor cells appeared exclusively in the photoreceptor cell layer 12 h after ornithine administration. The number of TUNEL-positive cells increased throughout the 2 days following the injection, then decreased markedly. TUNEL-positive cells remained until 28 days, when the photoreceptor cells had disappeared. The ganglion cell layer, inner nuclear layer and damaged RPE cells were negative for TUNEL staining during all stages. The electron microscopic study also revealed the pyknotic nuclei of apoptotic photoreceptor cells. CONCLUSION: An intravitreal injection of ornithine caused primary damage to the RPE, and subsequently some of the photoreceptor cells revealed apoptosis by TUNEL assay. These findings suggest the dysfunction of the RPE causes photoreceptor cell death according to the intrinsic program of an apoptotic mechanism.

Animals↗

PubMed1997

Interleukin-2 modulates the responsiveness to angiotensin II in cultured vascular smooth muscle cells.

Preincubation with interleukin-2 (IL-2), a T cell-derived cytokine, enhanced the increase in intracellular Ca2+ ([Ca2+]i) induced by angiotensin II (AII) in vascular smooth muscle cells (VSMC). IL-2 itself did not affect the basal [Ca2+]i level or the maximal response of [Ca2+]i increase induced by AII. Furthermore, IL-2-induced enhancement was not observed in the absence of extracellular Ca2+, suggesting that IL-2 enhances Ca2+ influx induced by AII. IL-2 also enhanced the stimulation of DNA synthesis induced by AII, although IL-2 alone did not stimulate DNA synthesis. Genistein, an inhibitor of protein tyrosine kinases, significantly inhibited IL-2-induced enhancement of both Ca2+ influx and DNA synthesis induced by AII. A neutralizing antibody against heparin-binding epidermal growth factor-like growth factor (HB-EGF) partially inhibited IL-2-induced enhancement of DNA synthesis induced by AII. These findings suggest that autocrine HB-EGF is partially involved in the mechanism of IL-2-induced enhancement of DNA synthesis. On the other hand IL-2 stimulated both glycosaminoglycan (GAG) and prostacyclin syntheses and enhanced the stimulation of both GAG and prostacyclin syntheses induced by AII. Therefore, IL-2 may play important roles in the pathogenesis of atherosclerosis and vascular disease by modulating the responsiveness to AII in VSMC.

Angiotensin II↗

PubMed1997

Coil embolization of ruptured vertebral dissection in acute stage with interlocking detachable coils.

BACKGROUND: Although dissecting aneurysm of vertebral artery is known as one of the causes of subarachnoid hemorrhage (SAH) in the posterior circulation, the best surgical treatment method remains controversial. METHOD AND RESULT: This 64-year-old woman was admitted to our service with headache due to SAH caused by a ruptured vertebral dissecting aneurysm in the distal portion of the posterior inferior cerebellar artery. After confirming tolerance of parent artery occlusion by temporary balloon occlusion, both the dissection site and the proximal portion of the parent artery were occluded completely by interlocking detachable coils (IDCs) without any ischemic complications. The patient was discharged without any neurologic deficit on the 25th day after the therapy. CONCLUSION: The goal of treatment for the ruptured dissecting aneurysm is isolation of the dissection site from the circulation to prevent rerupture. In our case, endovascular occlusion with IDCs was sufficient to reach the goal. In cases with difficulties in the surgical approach, embolization of the dissection site with IDCs should be considered.

Aneurysm, Ruptured↗

PubMed1997

Morphological changes of intraparenchymal arterioles after experimental subarachnoid hemorrhage in dogs.

OBJECTIVE: Morphological and microcirculatory changes in intraparenchymal vessels after subarachnoid hemorrhage (SAH) have not yet been fully clarified. We conducted this experimental study to investigate the serial morphological changes of intraparenchymal arterioles after SAH. METHODS: SAH was produced by injecting autologous arterial blood into the cisterna magna twice at 48-hour intervals in 30 dogs. The dogs were killed 3, 7, or 14 days after SAH, and then perfusion-fixed specimens of both anterior sylvian giri were obtained by using two methods. Microvascular corrosion casts produced by arterial injection of polyester resin were examined using scanning electron microscopy, and the widths of 40 arterioles of each animal were measured. Sectioned slices from the brain surface to 500 microns deep were examined by light microscopy, and external diameter, internal diameters, and wall thickness of the arterioles at depths of 50, 200, and 500 microns from the brain surface were morphometrically evaluated in 40 arterioles of each animal. In control animals receiving cisternal injections of mock cerebrospinal fluid (n = 10) and in healthy control animals (n = 10), the same examination and evaluation were performed. RESULTS: Corrosion casts of arterioles showed tapered narrowing with folding after SAH, and the width of the arterioles significantly decreased 3 and 7 days after SAH (P < 0.01). Morphometric examination by light microscopy showed a significant decrease of internal diameter of arterioles associated with a significant increase of wall thickness at any depth from the brain surface 3 and 7 days after SAH (P < 0.05 or P < 0.01). These findings improved 14 days after SAH. Control animals receiving cisternal injections of mock cerebrospinal fluid showed no significant differences compared with healthy control animals. CONCLUSION: These results suggest that constriction of intraparenchymal arterioles occurs after SAH and may contribute to delayed cerebral ischemia.

Animals↗

PubMed1997

Test-retest and inter-method reliability of the self-rating Barthel Index.

OBJECTIVE: To develop a self-rating Barthel Index questionnaire (SB) for assessing stroke patients living at home, the test-retest reliability of SB versions 1 and 2 and the inter-method reliability of SB version 3 were examined. DESIGN: Case series. SETTING: Clinics of the Department of Rehabilitation Medicine at the University Hospital and two affiliated hospitals. PATIENTS: Thirty-one, 140 and 65 chronic stroke patients living at home, being seen for regular follow-up, and having no severe aphasia for SB versions 1, 2 and 3, respectively. MAIN OUTCOME MEASURE: Kappa coefficients of items in SB versions 1 and 2 for test-retest reliability, intra-class correlation coefficients of the self-care index, mobility index and total score, kappa coefficients of items and Friedman's two-way ANOVAs of ranking in SB version 3 among the self-ratings, family ratings and occupational therapist (OT) ratings; a regression analysis of age, gender, duration, time for filling in the SB, OT ratings, Mini-Mental State score and aphasia. RESULTS: Kappa coefficients in SB version 2 were higher than those in version 1. Intra-class correlation coefficients in SB version 3 among the three ratings were good, and kappa coefficients in each item were also good or excellent. By Friedman's two-way ANOVA, all the items except walking up/down stairs had no significant difference in ranking. Regression analysis revealed that OT ratings and time for filling in the SB affected the absolute difference of scores between the self-ratings and OT ratings. CONCLUSIONS: SB version 3 is sufficiently reliable for practical use.

Activities of Daily Living↗

PubMed1997

[Indocyanine green infrared fluorescence angiography and histopathological correlation in experimental choroidal circulatory disturbance. Report 1].

We performed an experimental study on choroidal circulatory disturbance to clarify basic problems about interpretation of retino-choroidal lesions in indocyanine green fluorescence angiography (ICG angiography). We severed all of the posterior temporal ciliary arteries, to produce choroidal circulatory disturbance. Fluorescein angiography and ICG angiography were performed using scanning laser ophthalmoscope immediately, and 2 days after occlusion. These findings were compared with histopathological findings from the same specimen. Immediately after occlusion, choroidal vessels were filled with the red blood cells in the lesion that showed hypofluorescence in both types of angiography. Two days after occlusion, the fundus had a grayish white edematous appearance which was similar to choroidal infarction. The retinal pigment epithelial cells. (RPEs) in infarcted lesion progressed to liquefied necrosis. Fluorescein angiography showed hyperfluorescence in the lesion, and ICG angiography showed hypofluorescence in the early phase, but hyperfluorescence at the margin of the lesion in the late phase. This result showed that damaged RPEs were stained by ICG dye. In reading ICG angiography, we have to consider that the ICG angiogram is greatly modified by the condition of the RPEs.

Animals↗

PubMed1997

[Ornithine induced retinopathy in the rat--1. Histopathological study on the retinal pigment epithelium].

An intravitreal injection of a small amount of 1-ornithine hydrochloride damages the retinal pigment epithelium (RPE) selectively in monkey eyes. In this paper, we examined the rat eye and observed similar results. Clinically, retinal edema appeared immediately after injection and showed hyperfluorescence on a fluorescein angiograph after 3 days. Histopathologically, RPE showed marked swelling 3 hours after injection, then became necrotic and disappeared gradually. One month after injection, retinal structure was maintained in the area where RPE regenerated to cover Bruch's membrane, but in the area where RPE cells disappeared, the photoreceptor cells and choriocapillaris degenerated and finally disappeared. In the eyes injected intravitreally with NaCl solution of equivalent osmosis microvacuoles were seen but no necrotic change in RPE. Thus, a small amount of ornithine damaged RPE selectively, and induced disappearance of the photoreceptors and choriocapillaris secondarily.

Animals↗

PubMed1997

[Indocyanine green infrared fluorescence angiography and histopathological correlation in experimental choroidal circulatory disturbance. Report 2].

We performed an experimental study on choroidal circulatory disturbance to clarify basic problems about interpretation of retino-choroidal lesions in indocyanine green fluorescence angiography (ICG angiography). We severed the posterior ciliary arteries to produce choroidal circulatory disturbance. Fluorescein angiography and ICG angiography were performed at one week, and one month after occlusion. These findings were compared with histopathological findings. One week after occlusion, the area of choroidal infarct showed occlusion of choriocapillaris and proliferation of the retinal pigment epithelial (RPE) cells, this area showed hypofluorescence in the early phase ICG angiography. The hypofluorescence area increased in the late phase. One month after occlusion, the lesion showed loss of choriocapillaris at the center and proliferation of fibroblast-like cells at the edge of the lesion. The subretinal strand showed hyperfluorescence in late phase ICG angiography. Proliferated RPE cells masked ICG fluorescence in the late phase. Fibroblast-like cells showed tissue staining. When reading ICG angiography, we have to take into account that the ICG angiogram is greatly modified by condition of the RPE.

Animals↗

PubMed1997

[Ornithine induced retinopathy in rat--2. Process of disappearance of choriocapillaris].

We observed the process of disappearance of the choriocapillaris after loss of the retinal pigment epithelium (RPE) induced by intravitreal injection or ornithine. Three hours after administration of ornithine, the RPE cells swelled remarkably in the posterior pole, but, the endothelial cells of the choriocapillaris remained intact. At 3 days, the RPE cells became necrotic, but the choriocapillaris still preserved its in normal appearance. At 7 days, RPE disappeared completely in the posterior pole and the choriocapillaris displayed evidence of atrophy; the swollen lumen of the choriocapillaris became narrow the cytoplasm of the endothelium was swollen, and the number of fenestrae was reduced. On the other hand, these changes were not seen where the RPE remained. At 14 days, in the posterior pole, the lumen of the choriocapillaris occluded by the swollen endothelial cells. At 28 days, the choriocapillaris completely disappeared and the large choroidal vessel was directly in contact with Bruch's membrane. These results showed that the RPE is correlated with the presence of the choriocapillaris.

Animals↗

PubMed1996

[Preventive effect of intracisternal methylprednisolone on symptomatic vasospasm after aneurysmal subarachnoid hemorrhage].

A series of 103 patients with ruptured cerebral aneurysms were treated by intracranial irrigation, after aneurysmal clipping, with pH 8.0 Hartmann solution containing 1 mg/ml of methylprednisolone sodium succinate, during operation. Postoperatively, 5 ml/day of the solution was also injected until day 14, percutaneously through an Ommaya reservoir into the cisterns around the clipped aneurysm. Six (5.7%) of the 103 patients suffered symptomatic vasospasm postoperatively. The Glasgow Outcome Scale at discharge of all patients indicated good recovery, Moderately disabled, Severely disabled and Vegetative state in 74 (72%), 18 (17%), 9 (9%) and 2 (2%) patients respectively. As a complication, subdural effusion was noticed in 17 (17%) patients, without any influence on outcome at all.

Female↗

PubMed1996

[Indocyanine green infrared fluorescence angiography and vascular cast--preparation in experimental choroidal circulatory disturbance].

We performed experiments in 20 monkey eyes in order to clarify basic problems about interpretation of indocyanine green fluorescence angiography (ICG angiography). We severed the temporal group of posterior ciliary arteries to produce choroidal circulatory disturbance. ICG angiography was performed immediately, and 2 days, 4 days, and 2 weeks later. Following each ICG angiography, the eye was studied by plastic vascular cast technique with scanning electron microscopy. Immediately after occlusion, ICG angiography showed filling defect in the temporal choroidal hemisphere during the early phase. In the later phase, this area was gradually filled by the dye from choroidal arteries in the nasal hemisphere and the anterior ciliary arteries. Vascular cast preparations showed filling defect in the temporal choroidal hemisphere, corresponding with the early ICG angiogaphic findings. Both filling delay in ICG angiography and filling defect in vascular casts improved daily after occlusion. Two weeks after occlusion, The area of choroidal infarct temporal to the macula turned into chorioretinal atrophy. This area showed hypofluorescence in the early-phase ICG angiography and filling defect of the choriocapillaris in plastic casts. The early-phase ICG angiographic findings thus corresponded well with observations of vascular casts. We conclude that ICG angiography correctly reflects the actual circulatory disturbances in the choroid.

Animals↗

PubMed1996

[Indocyanine green angiography of choroidal neovascularization in age-related macular degeneration].

One hundred forty eyes with choroidal neovascularization (ChNV) in age-related macular degeneration were examined with fluorescein and indocyanine green (ICG) angiography. ChNVs covered with subretinal hemorrhage, and ChNVs with retinal pigment epithelial (RPE) detachment were demonstrated at a higher rate and more clearly with ICG angiography than with fluorescein angiography. On the other hand, ChNVs in serous retinal detachment, and in disciform lesions were demonstrated more clearly with fluorescein angiography than with ICG angiography. These results show that ICG angiography is valuable for delineating "occult ChNVs" with subretinal hemorrhage or RPE detachment in age-related macular degeneration.

Aged↗

PubMed1995

High-resolution solution structure of siamycin II: novel amphipathic character of a 21-residue peptide that inhibits HIV fusion.

The 21-amino acid peptides siamycin II (BMY-29303) and siamycin I (BMY-29304), derived from Streptomyces strains AA3891 and AA6532, respectively, have been found to inhibit HIV-1 fusion and viral replication in cell culture. The primary sequence of siamycin II is CLGIGSCNDFAGCGYAIVCFW. Siamycin I differs by only one amino acid; it has a valine residue at position 4. In both peptides, disulfide bonds link Cys1 with Cys13 and Cys7 with Cys19, and the side chain of Asp9 forms an amide bond with the N-terminus. Siamycin II, when dissolved in a 50:50 mixture of DMSO and H2O, yields NOESY spectra with exceptional numbers of cross peaks for a peptide of this size. We have used 335 NOE distance constraints and 13 dihedral angle constraints to generate an ensemble of 30 siamycin II structures; these have average backbone atom and all heavy atom rmsd values to the mean coordinates of 0.24 and 0.52 A, respectively. The peptide displays an unusual wedge-shaped structure, with one face being predominantly hydrophobic and the other being predominantly hydrophilic. Chemical shift and NOE data show that the siamycin I structure is essentially identical to siamycin II. These peptides may act by preventing oligomerization of the HIV transmembrane glycoprotein gp41, or by interfering with interactions between gp41 and the envelope glycoprotein gp120, the cell membrane or membrane-bound proteins [Frèchet, D. et al. (1994) Biochemistry, 33, 42-50]. The amphipathic nature of siamycin II and siamycin I suggests that a polar (or apolar) site on the target protein may be masked by the apolar (or polar) face of the peptide upon peptide/protein complexation.

Amino Acid Sequence↗