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Our previous study showed that acute restraint stress enhances depolarization-induced increases in intrasynaptosomal free calcium (Ca(2+)) concentration ([Ca(2+)](i)) and Ca(2+)-dependent glutamate release in mouse cerebrocortical nerve terminals (synaptosomes). In the present study, we investigated the effects of chronic stress on [Ca(2+)](i) and glutamate release in cerebrocortical synaptosomes from mice.Male ddY strain mice were randomly assigned to one of two experimental groups: control group and chronic stressed group. Mice in the chronic stressed group were subjected to immobilization stress for 2 hours daily for a period of 21 days. [Ca(2+)](i) and glutamate release in cerebrocortical synaptosomes isolated from the mice were determined by fura-2 fluorescence assay and enzyme-linked fluorometric assay, respectively.Chronic stress caused a significant increase in resting [Ca(2+)](i) and significantly enhanced the ability of the depolarizing agents K(+) and 4-aminopyridine (4-AP) to increase [Ca(2+)](i). It also brought about a significant increase in spontaneous (unstimulated) glutamate release and significantly enhanced K(+)- and 4-AP-evoked Ca(2+)-dependent glutamate release. Synaptosomes were more sensitive to the depolarizing agents at lower concentrations following chronic stress than after acute stress. The pretreatment of synaptosomes with a combination of omega-agatoxin IVA (a P-type Ca(2+) channel blocker) and omega-conotoxin GVIA (an N-type Ca(2+) channel blocker) completely suppressed the enhancements of [Ca(2+)](i) and Ca(2+)-dependent glutamate release in chronic stressed mice.These results indicate that chronic stress enhances depolarization-evoked glutamate release by increasing [Ca(2+)](i) via stimulation of Ca(2+) entry through P- and N-type Ca(2+) channels, and that chronic stress increases the sensitivity to depolarizing agents.

A 61-year-old woman was admitted for head injury after a traffic accident. Two months later, she developed abducens nerve palsy, chemosis, and pulsatile tinnitus. Right internal carotid angiography demonstrated a high flow direct carotid-cavernous fistula (CCF) at the C(5) portion with reflux into the superficial and deep sylvian veins, superior ophthalmic vein, superior petrosal sinus, and inferior petrosal sinus. Intravascular ultrasonography (IVUS) revealed a large fistula at the C(5) portion of the internal carotid artery (ICA). Coil embolization via transarterial and transvenous approaches under IVUS monitoring was performed. During the procedure, IVUS accurately detected protrusion of a coil into the parent ICA, and the parent artery could be preserved. IVUS monitoring is useful for embolization of direct CCF with coils.

Genes involved in the fertility restoration (Rf) of wild-abortive (WA)-type cytoplasmic male sterility (CMS) in rice have not been clearly identified, because of the inconsistency of their number and map position. Although the Rf-1 genes, which restore Chinsurah Boro (BT)-type CMS, have been recently isolated, their effects on WA-type CMS are still unclear. In the present study, near-isogenic lines (NILs) of japonica rice, differing at Rf loci, were developed using marker-assisted selection (MAS) to determine the effect of each Rf gene on WAor BT-cytoplasm in a japonica background. The NILs were crossed with a japonica CMS line, then the pollen and seed fertility of the hybrid plants was analyzed. The Rf-1 locus consisting of two Rf genes from IR36 was sufficient to restore fertility in the hybrids with BT-type CMS but conferred very little fertility to those with WA-type CMS. Rf-3 on chromosome 1, a weak Rf gene, and Rf-6(t) on chromosome 10 were unable to confer seed fertility to plants with BT-type or WA-type CMS. Both Rf genes, however, could accelate the development of abortive pollen in the case of WA-type CMS and their effect was additive. Crossing of CMS rice with chromosome segment substitution lines carrying Rf loci from IR24 resulted in similar pollen development and seed fertility.

Using a functional polymer containing nitrospirobenzopyran residues and poly(ethylene glycol), we developed a photoresponsive cell culture surface (PRCS), at which the cell adhesion can be enhanced locally by UV light irradiation (350–400 nm, 35 mW/cm2, 5 min) even on the region where living cells exist. Cell adhesion was evaluated by observing BALB/3T3 fibroblasts, which had been seeded on PRCS and then irradiated with UV light regionally, after 12 h incubation and subsequent washing to remove scarcely adhering cells. As a result, it was confirmed that the number of remaining cells in irradiated region was at most 2.5 times greater than that in nonirradiated regions, suggesting the implementation of a novel scheme to manipulate living cells individually by light irradiation in a parallel and simultaneous manner. Influence of the composition of the polymer material consisting the substrate surface was also investigated systematically. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 495–499, 2006

A 48-year-old woman was admitted to our emergency center for deterioration of consciousness. On arrival, her level of consciousness was 3 by the Japan Coma Scale and 10 by the Glasgow Coma Scale. Her blood pressure was 144/100 mmHg, and her pulse was regular at 136 beats/min. She was intubated under general anesthesia to lower the systolic blood pressure to 90-100 mmHg. Laboratory analysis showed elevated blood glucose (329 mg/dl), lactic dehydrogenase (311 U/l), creatinine kinase (CK, 410 U/l), and white blood cell count (19600/μl), and a reduced serum potassium (3.1 mmol/l). A computed tomographic scan showed a diffuse thick subarachnoid hemorrhage (SAH), and a chest X-ray revealed cardiomegaly and pulmonary edema. Electrocardiography on admission demonstrated sinus tachycardia and ST segment elevation in leads I, II, aVL, and V3 to V6, and a slightly prolonged QTc interval of 0.45 second. Echocardiography showed that wall motion of the left ventricular apex was significantly reduced, indicating specific “takotsubo” cardiomyopathy. Her cardiac function appeared to be well tolerant for direct surgery. Digital subtraction angiography was performed and disclosed an aneurysm at the P1 segment of the left posterior cerebral artery. Surgical intervention was undertaken by the left pterional approach. While a subarachnoid clot in the prepontine cistern was being evacuated to expose the aneurysm via an opticocarotid triangle, her blood pressure suddenly dropped to 20 mmHg, her heart rate declined to 20 beats/min, and carotid pulsation disappeared. The surgery was discontinued and cardiac resuscitation was conducted by intravenous injection of 1 mg epinephrine and cardiac massage by chest compression. Twenty-five minutes later her cardiac function stabilized, and the aneurysm was successfully clipped. The postoperative course was uneventful, and the electrocardiogram showed inversion of T waves and gradual improvement of echocardiographic abnormalities. An excessive catecholamine secretion related to hypothalamic damage is presumed to be the cause of cardiac dysfunction in the acute phase of SAH. Although evacuation of prepontine clot is a routine surgical maneuver, subtle compression or traction of the perforators from the P1 segment that feeds the hypothalamus might further injure the already affected hypothalamus to result in sudden cardiac arrest. In treatment of patients with SAH associated with cardiac dysfunction, preparation of an intra-aortic balloon pumping would be beneficial.

重症例に対しても待機せず可及的速やかに脳動脈瘤根治術を行うという方針のもと診断治療を行った,1996年1月から2000年12月までの5年間の235名のクモ膜下出血(SAH)患者につき検討した。患者の内訳は男性85名,女性150名,平均年齢60.8歳であった。来院時WFNS Grade (G)はGI 21名,GII 48名,GIII 2名,GIV 51名,GV 79名,および来院時心肺機能停止(CPA): 34名であり,GIV, GV, CPAが全体の70%を占めた。原則としてGIからGIVまでの全例,GVでは脳幹反射がみられるものを脳動脈瘤根治術の対象とし,待機せず直ちに診断,治療を開始した。CPAで早期に心肺蘇生が施行され自己心拍の再開をみたものはGVに準じた。167名(71.1%)に脳血管撮影(DSA), 146名(62.1%, 8名3.4%はDSA未施行)にクリッピング術,19名(8.1%)にGuglielmi detachable coil瘤内塞栓術を施行した。発症からの時間経過(中央値)は,来院まで47分,脳血管撮影まで200分,手術まで338分であった。1998年以降,21名に脳低温療法を行った。6か月後のGlasgow outcome scaleによる全体の転帰は,Good recovery (GR): 67名(29.5%), moderate disability (MD): 19名(8.4%), severe disability (SD): 38名(16.7%), persistent vegetative state (VS): 10名(4.4%), death (D): 93名(41.0%)であった。軽症例(GI, II, III)の転帰良好(GR+MD)は78.3%,不良(SD+VS) 10.1%,死亡11.6%,重症例(GIV, V)の転帰良好は24.8%,不良31.2%,死亡44%であった。CPAから蘇生後の4例に根治術施行,3名が生存,うち1名はGRであった。重症SAH症例が過半数を占める救命救急センターにおいて,“救いうる”SAH患者に一人でも多く良好な転帰を得るためには,来院後の再破裂予防と迅速な根治術施行とともに,低酸素脳障害の予防を含め,病院外救護体制の面からも取り組みが必要と思われた。

We retrospectively analyzed and compared overall outcome of poor-grade patients (WFNS Grade IV, V, and cardiopulmonary arrest on arrival |CPAOA|) with subarachnoid hemorrhage (SAH) in 2 study periods. The first was from July 1993 to June 1994 and the second from July 1998 to June 1999. A total of 20 patients, 7 with Grade IV, 5 with Grade V, and 8 with CPAOA were included in the first period; and a total of 36 patients, 6 with Grade IV, 21 with Grade V, and 9 with CPAOA, in the second period. The major recent changes in the treatment protocol were 1) deep sedation, intensive control of blood pressure, and endotracheal intubation in the emergency room in the resuscitative phase; 2) immediate cerebral angiography under general anesthesia after diagnosis of SAH; 3) aneurysm obliteration by Guglielmi detachable coil (GDC) as an option; and 4) early enteric feeding and tracheostomy for patients in a prolonged comatose state. Therapeutic mild brain hypothermia was conducted in 3 Grade V patients in the second-period study. Aneurysm was obliterated by clipping in 7 and 18 patients in each period and by GDC in 4 patients in the second period. The median time from onset to admission, to angiography, and to surgery was 29, 440, and 797.5 minutes, respectively, in the first period, and 32, 99, and 210 minutes, respectively, in the second period. Overall outcome assessed at 3 months by the Glasgow Outcome Scale was; GR: 2 (10%), MD: 0 (0%), SD: 1 (5%), VS: 2 (10%), D: 15 (75%) for the first period and GR: 5 (13.9%), MD: 3 (8.3%), SD: 7 (19.4%), VS: 1 (2.8%), D: 20 (55.6%) for the second period. A favorable outcome (GR+MD) increased from 10% to 22.2%, and deaths were reduced from 75% to 55.6% (P=0.02, chi-square test). Angiography and surgical intervention immediately after resuscitation produce a more favorable outcome for severe SAH patients without increasing the risk of rerupture.

A 77-year-old woman suddenly lost consciousness and presented with right hemiparesis. Computed tomography showed a large subcortical hematoma in her left frontal lobe associated with subarachnoid hemorrhage. The first impression was hemorrhage due to a ruptured aneurysm of the middle cerebral or the internal carotid artery on the left. Left internal carotid angiography showed no aneurysm or vascular anomaly, but back flow of contrast medium into the external carotid artery disclosed two saccular aneurysms arising from the anterior branch of the left middle meningeal artery (MMA). Emergent surgical intervention confirmed that the hematoma was due to ruptured MMA aneurysm. Nontraumatic MMA aneurysm should be recognized as a possible causative lesion of intracranial hemorrhage despite the extremely low incidence. External carotid angiography should be performed in patients with intracranial hemorrhage in whom no vascular cause can be detected in the internal carotid system.

最重症くも膜下出血(SAH)に対する可及的早期脳低温療法の有用性の検討するため,1999年4月以降に経験した75歳未満のWFNS Grade VのSAH 16名に診断確定直後より脳低温療法を施行し,脳動脈瘤根治術を施行した。内訳は男性10名,女性6名,年齢50-72歳(平均63.1歳),Glasgow Coma Scale (GCS)は,心肺停止から蘇生しGCS 3となったもの2名,3が6名,4が1名,5が3名,6が4名であった。CT上4名が10mm以上の正中偏位を伴う血腫を合併し,4名が鋳型状脳室内血腫を伴った。SAHの診断確定後,速やかに脳低温療法を開始し,クリッピング術,外減圧術を施行した。経過時間(中央値)は発症後搬入まで33分,脳血管撮影まで86分(7名は3D-CTAのみ施行),手術まで174分であった。33-34℃の脳低温療法を2-3日施行後,復温した。復温終了まで平均8.3日であった。発症24時間以内に経腸栄養を開始,穿刺式気管切開を行い肺理学療法を徹底した。脳低温療法施行16名のGlasgow Outcome Scaleによる転帰はgood recovery (GR) 2名,moderate disability (MD) 3名,severe disability (SD) 8名,persistent vegetative state (VS) 2名,death (D) 1名であった。脳低温療法の合併症による症状の悪化や中断例はなく,一次性脳損傷が転帰不良の原因と考えられた。脳低温療法導入直前のGrade VのSAH連続24名と導入後,脳低温の適応外となった6名を含む連続22名を比較すると,転移良好(GR+MD)率に関しては有意差は認めなかったが(12.5% vs. 22.7%), VS+Dの率は71.8%から29.2%と有意に低下した。最重症SAHに対する超急性期からの脳低温療法は少なくとも植物症への移行と死亡を減少させ,一次性脳損傷を軽減させる可能性がある。

Polymerase chain reaction (PCR) is an efficient method for obtaining a desired nt sequence if both required primers can hybridize to the target DNA molecule specifically. A rapid and simple PCR-based method for analyzing plasmids using intact cells was established. An attempt to target a rice waxy sequence by PCR using homologous primers was also carried out. In three cases, specific fragments were amplified and their nucleotide sequences were determined. However, the cloned rice waxy gene contained base substitution mutations. The cumulative frequency of mutation after 30 polymerization cycles was estimated to be one in 500 bp.

A simulation study was undertaken using a compartmental flow model of a large high-flow cerebral arteriovenous malformation to investigate the hemodynamic changes during obliteration procedures. Under certain autoregulatory conditions, marked hyperperfusion (92 ml/100 g/min) could be induced in association with increased wall stress of the arterioles. Narrowing of the autoregulatory pressure range and its shift to a low pressure level are suspected to be among the possible causes of normal perfusion pressure breakthrough phenomenon.

A photoresponsive culture surface (PRCS) allowing photocontrol of cell adhesion was prepared with a novel polymer material composed of poly(N-isopropylacrylamide) having spiropyran chromophores as side chains. Cell adhesion of the surface was drastically enhanced by the irradiation with ultraviolet (UV) light (wavelength: 365 nm); after subsequent cooling and washing on ice, many cells remained in the irradiated region, whereas most cells were removed from the nonirradiated region. The cell adhesion of the PRCS, which had been enhanced by previous UV irradiation, was reset by the visible light irradiation (wavelength 400-440 nm) and the annealing at 37 degrees C for 2 h. Also it was confirmed that the regional control of cell adhesion was induced several times by repeating the same series of operations. Further, living cell patterning with the 200 microm line width was produced readily by projecting UV light along a micropattern on the PRCS on which the living cells had been seeded uniformly in advance. By using a fluorescent probe that stains living cells only, it was confirmed that the cells maintained sufficient viability even after UV light irradiation followed by cooling and washing.

Little has been reported on the flow volume of cortical arteries in the posterior circulation. During craniotomy in 28 patients, we measured the flow velocity of the arteries using microvascular Doppler sonography and recorded their arterial radii and the insonation angle. The flow velocities and radii were 25.7 cm sec-1 and 0.11 cm for the posterior cerebral artery (PCA), 19.5 cm sec-1 and 0.07 cm for the superior cerebellar artery (SCA), 19.6 cm sec-1 and 0.04 cm for the anterior inferior cerebellar artery (AICA), 14.9 cm sec-1 and 0.05 cm for the posterior inferior cerebellar artery (PICA), 28.3 cm sec-1 and 0.18 cm for the basilar artery, and 18.5 cm sec-1 and 0.16 cm for the vertebral artery, respectively. Since the flow velocities of these arteries were confined in a narrow range compared to the cross-sectional areas, the latter was considered to be the primary determinant for flow volume in these arteries. Based on certain assumptions, we estimated the flow volume of the PCA, SCA, AICA and ICA to be 60, 20, 10 and 10 ml min-1, respectively, which could be regarded as the current reference values for the arterial flow volumes.

Hemodynamics of paraclinoid aneurysms were investigated focusing on the effects of their locations and shape of the carotid siphon. A transparent silicon model of the carotid siphon was constructed and a model aneurysm was attached to the outside of the curvature at three different sites. Glycerol solution was perfused into the model, and the half-life of the dye injected into the aneurysm was calculated as an index of the stagnant flow. Values of half-life changed significantly depending on the aneurysmal location and the siphon angle. When the siphon angle was 0 degree where C2 and C4 segments were parallel to each other, the half-life value was the lowest in the C2-C3 junction aneurysm, highest in the C3 segment aneurysm and intermediate in the C2 segment aneurysm. While the C2-C3 junction aneurysm maintained low values regardless of the angle, the C3 segment aneurysm values decreased and C2 segment aneurysm values increased with increases in the angle. These changes of half-life occur because the point at which the faster moving fluid component strikes the curved wall changes according to the siphon angle. These results are considered useful to determine the surgical indications, treatment modalities and post-surgical management of the aneurysms.

We simulated parent artery occlusion therapy for basilar head aneurysms to elucidate the hemodynamic changes induced by different occlusion sites and diameters of the posterior communicating arteries (PCom) as collateral pathways. A vascular model of the vertebrobasilar system with a basilar head aneurysm was constructed. Four types of occlusion were simulated: Basilar artery occlusion distal to (Type A), between (Type B) and proximal to (Type C) the superior cerebellar arteries, and bilateral vertebral artery occlusion (Type D). Glycerol solution was perfused into the model, and the half-life of the dye injected into the aneurysm was calculated and regarded as an index of stagnant flow in the aneurysm. The half-life was increased significantly and nonlinearly after parent artery occlusion, depending on the occlusion site and the ratio of two PCom diameters (diameter ratio). Intra-aneurysmal stagnation developed markedly in Type A and Type B in the diameter ratio higher than 0.70 and considerably in Type C in the ratio higher than 0.80. Additional P1 occlusion of the posterior cerebral artery enhanced the stagnation in Type A and B. Since the results are consistent with the published clinical data, the simulation study will be useful for speculating the efficacy of the therapeutic occlusion.

Hemodynamic changes following unilateral vertebral artery (VA) occlusion were investigated in a rat model. The left carotid artery was resected and anastomosed to the right side in an end-to-side fashion to create a half-ring bypass. The distal side of the bypass was regarded as a union of VAs. Changes in the geometry, histology and hemodynamics in the union were investigated after the recipient artery was ligated. Intimal thickening was most prominently observed in the recipient arterial segment distal to the ligation site, where the lumen was obliterated. However, the portion of the lumen within 2.6 +/- 0.6 (mean +/- s.d.) mm, a distance of 2.9 +/- 0.6 times the internal diameter from the union, was not obliterated. The angle of the union was positively related to the length of this residual lumen. The results of this study explain some of the pathogenesis in unsuccessful aneurysmal thrombosis or brain stem infarction after therapeutic unilateral VA occlusion.