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A simulation model was developed to better understand the mechanisms of brain injuries in sports. A three-dimensional model comprising approximately 1.22 million elements was constructed from cranial computed tomography images of adult male volunteers by the voxel method. To simulate contact sports that permit actions such as tackling, a sinusoidal wave with duration of 10 ms and maximum acceleration of 2000 m/s2 was applied to the lowest point of the model to apply rotational acceleration to the head from different directions. The von Mises stress was then observed at five points in the coronal plane of the brain: cingulate gyrus (CG), corpus callosum (CC), brain stem (BS), lateral temporal lobe (LT), and medial temporal lobe (MT). LS-DYNA universal finite element analysis software with explicit time integration was used for the analysis. Concentrations of stress started to appear in the CC and BS at 10 ms post-impact, after which they also became evident in the CG and MT. The maximum changes in stress at each location occurred 10-15 ms post-impact. The von Mises stress was 9-14 kPa in the CG, 8-24 kPa in the CC, 12-24 kPa in the BS, 7-12 kPa in the LT, and 12-18 kPa in the MT. The highest stress in every part of the brain occurred after lateral impact, followed by oblique impact and sagittal impact. Such simulations may help elucidate the mechanisms of brain injuries in sports and help develop measures to prevent chronic traumatic encephalopathy.

To screen for the 328 bp Alu insertion (c.4052_4053ins328, p.Tyr1352Alafs) in RP1 in a group of retinitis pigmentosa (RP) patients who had been previously identified with a heterozygous deleterious mutation in the gene. Prospective, clinical and experimental study. The Alu insertion in RP1 was screened with an optimized PCR-based method in 26 RP patients with a heterozygous deleterious mutation (nonsense or frameshift) in RP1 that had been identified in a preceding genetic study. The genetic location of the previously identified mutation and its inheritance pattern were assessed. Out of 26 RP patients with a heterozygous deleterious mutation in RP1, 5 (19.2%) were found to carry an additional heterozygous Alu insertion, presumably resulting in a compound heterozygous state. This included 3 patients who had been previously diagnosed as autosomal dominant RP based on genetic findings. They were re-diagnosed as having an autosomal recessive disease following our new findings. In all patients identified with the Alu insertion, the other mutations found in the preceding study were outside the defined region in exon 4 (encoding amino acids 677 to 917) in which truncation mutations have been suggested to exert a dominant negative effect. The founder Alu insertion in RP1 is an important cause of autosomal recessive RP in Japanese patients and can be missed in standard targeted resequencing. Screening optimized for this mutation is warranted, particularly in patients with a heterozygous deleterious mutation outside the defined region in exon 4 of RP1.

BackgroundThe genetic profile of retinitis pigmentosa (RP) in East Asian populations has not been well characterised. Therefore, we conducted a large-scale sequencing study to investigate the genes and variants causing RP in a Japanese population.MethodsA total of 1209 Japanese patients diagnosed with typical RP were enrolled. We performed deep resequencing of 83 known causative genes of RP using next-generation sequencing. We defined pathogenic variants as those that were putatively deleterious or registered as pathogenic in the Human Gene Mutation Database or ClinVar database and had a minor allele frequency in any ethnic population of ≤0.5% for recessive genes or ≤0.01% for dominant genes as determined using population-based databases.ResultsWe successfully sequenced 1204 patients with RP and determined 200 pathogenic variants in 38 genes as the cause of RP in 356 patients (29.6%). Variants in six genes (EYS, USH2A, RP1L1, RHO, RP1 and RPGR) caused RP in 65.4% (233/356) of those patients. Among autosomal recessive genes, two known founder variants in EYS [p.(Ser1653fs) and p.(Tyr2935*)] and four East Asian-specific variants [p.(Gly2752Arg) in USH2A, p.(Arg658*) in RP1L1, p.(Gly2186Glu) in EYS and p.(Ile535Asn) in PDE6B] and p.(Cys934Trp) in USH2A were found in ≥10 patients. Among autosomal dominant genes, four pathogenic variants [p.(Pro347Leu) in RHO, p.(Arg872fs) in RP1, p.(Arg41Trp) in CRX and p.(Gly381fs) in PRPF31] were found in ≥4 patients, while these variants were unreported or extremely rare in both East Asian and non-East Asian population-based databases.ConclusionsEast Asian-specific variants in causative genes were the major causes of RP in the Japanese population.

It is important to distinguish foramen magnum arachnoiditis (FMA) from Chiari malformation (CM) before surgery because the operative strategies for these diseases differ. In the current study, we compared pretreatment magnetic resonance imaging (MRI) of FMA with CM and investigated the MRI findings useful to differentiate between these diseases. We retrospectively reviewed patients with FMA or CM aged ≥ 18 years who underwent surgeries at our institution between 2007 and 2019. The morphologies of the syrinx, neural elements, and posterior cranial fossa were preoperatively evaluated with MRI. We used the receiver operating characteristic (ROC) curve for the fourth ventricle-to-syrinx distance (FVSD). Ten patients with FMAs and 179 with CMs were included. FVSD in the FMA group was significantly shorter than that in the CM group (7.5 mm [IQR, 2.8–10 mm] in FMA vs. 29.9 mm [IQR, 16.3–52.9 mm] in CM, p

PURPOSE To evaluate retinal vessel density and retinal sensitivity (RS) after macular hole surgery with the superior inverted internal limiting membrane flap technique. METHODS Retrospective, observational case series. Twenty-one patients with idiopathic macular hole underwent 27-gauge vitrectomy with the superior inverted internal limiting membrane flap technique and triamcinolone acetonide. Measurements included RS, which was measured with microperimetry, as well as retinal vessel density in the superficial capillary plexus (SCP) and deep capillary plexus (DCP), which was measured with optical coherence tomography angiography. All parameters were evaluated in the superior and inferior sectors of the macula preoperatively and 1, 3, and 6 months postoperatively. RESULTS Six months postoperatively, retinal thickness in the inferior sector was unchanged, but retinal thickness in the superior sector decreased significantly (P < 0.01). SCP vessel density in both sectors was unchanged at all postoperative time points. DCP vessel density in both sectors increased very significantly at 3 months (P < 0.01) and returned to baseline at 6 months. RS in the inferior sector increased by 47% 3 months postoperatively and by 61% 6 months postoperatively (P < 0.05 and P < 0.001, respectively), but RS in the superior sector increased only at 6 months postoperatively and only by 22% (P < 0.05). CONCLUSION Lower recovery of RS in the superior sector suggests that internal limiting membrane peeling might affect the postoperative visual function.

PURPOSE: Report global adalimumab safety and efficacy outcomes in patients with non-infectious uveitis.METHODS: Adults with non-infectious intermediate, posterior, or panuveitis were randomized 1:1 to receive placebo or adalimumab in the VISUAL I (active uveitis) or VISUAL II (inactive uveitis) trials. Integrated global and Japan substudy results are reported. The primary endpoint was time to treatment failure (TF).RESULTS: In the integrated studies, TF risk was significantly reduced (hazard ratio [95% CI]) with adalimumab versus placebo (VISUAL I: HR = 0.56 [0.40-0.76], p < 0.001; VISUAL II: HR = 0.52 [0.37-0.74], p < 0.001). In Japan substudies, no consistent trends were observed between groups (VISUAL I: HR = 1.20 [0.41-3.54]; VISUAL II: HR = 0.45 [0.20-1.03]). Adverse event rates were similar between treatment groups in both studies (854 to 1063 events/100 participant-years).CONCLUSIONS: Adalimumab lowered time to TF versus placebo in the integrated population; no consistent trends were observed in Japan substudies. Safety results were consistent between studies.

Rapid clearance and low ocular bioavailability are drawbacks of conventional ophthalmic eye drops. To increase the ocular drug resistance time and improve efficacy, an in situ forming and thermosensitive chitosan-gelatin hydrogel was developed. The feasibility of using this hydrogel as a topical eye drop formulation for sustained release of timolol maleate was evaluated. The flexible hydrogel that was co-crosslinked with β‑glycerophosphate disodium salt hydrate (β-GD) and genipin showed a fast gel formation at 37 °C. The swelling properties and in vitro biodegradation characteristics showed a strong relationship with the initial genipin concentration. In vitro release profiles demonstrated that crosslinking with genipin reduced the release rate of entrapped model drugs and timolol maleate. In vitro cytotoxicity tests showed that the hydrogel was non-toxic to Chinese hamster fibroblast V79 cells. The hydrogel was further applied as eye drop formulations for sustained release of timolol maleate to reduce intraocular pressure (IOP). A fast gel formation was observed after instilling the chitosan-gelatin solution into the lower conjunctival sac of the rabbit eyes, and the in situ formed hydrogels protected the drugs from clearance by tears, and released the drugs in a sustained manner. Furthermore, administration of timolol maleate containing chitosan-gelatin hydrogels showed a long-lasting and effective IOP lowering efficacy for up to 24 h compared with the conventional eye drops. These results suggested that β-GD and genipin co-crosslinked chitosan-gelatin hydrogels could be a useful ocular drug delivery platform with enhanced therapeutic effects and reduced side effects.

In this study, we developed a subcutaneous insulin-releasing device consisting of a disk-shaped capsule and drug formulation comprised of poly(ethylene glycol) dimethacrylates, then evaluated its efficacy on retinal function in streptozotocin (STZ)-induced diabetic rats. In vitro release studies showed that recombinant human insulin was released with a constant rate for more than 30 days. The device was able to maintain a basal level of blood glucose in diabetic rats for a prolonged period of more than 30 days, simultaneously preventing a decrease in body weight. For assessing the pharmacological effect of the device on retinal function in diabetic rats, electroretinograms were conducted for 12 weeks. The reduction in amplitude and delay in implicit time were attenuated by the device during the initial 4 weeks of application. The increase in gene expression of protein kinase C (PKC)-γ and caspase-3 in the diabetic retina was also attenuated by the device. Immunohistochemistry showed that the increase in glial fibrillary acidic protein expression in the diabetic retina was attenuated by the device. Histological evaluation of subcutaneous tissue around the device showed the biocompatibility of the device. In conclusion, the insulin-releasing device attenuated the reduction of retinal function in STZ-induced diabetic conditions for 4 weeks and the efficacy of the device might be partially related to PKC signaling in the retina. The long-term ability to control the blood glucose level might help to reduce the daily frequency of insulin injections.

To present phenotypic features of 22 patients with S-antigen (SAG) mutations.Retrospective cohort study.Twenty-one Japanese patients from 16 families with a homozygous c.924delA mutation and 1 patient with a homozygous c.636delT mutation in the SAG gene.Clinical records on symptoms; best-corrected visual acuity; and Goldmann perimetry, fundus photography, fundus autofluorescence (FAF), OCT, and electroretinography results were reviewed.Best-corrected visual acuity, Goldmann perimetry results, imaging findings, and electroretinography results.Ten patients had Oguchi disease and 12 had retinitis pigmentosa (RP) with mean follow-up periods of 13.8 and 10.2 years, respectively. Retinitis pigmentosa patients were older (mean age, 56.0 years) than those with Oguchi disease (mean age, 22.1 years; P0.001) at the initial visit. Night blindness noted in childhood was the most common initial symptom for both Oguchi disease (80.0%) and RP (91.7%) patients. Best-corrected visual acuity in the logarithm of the minimum angle of resolution (logMAR) was well preserved in Oguchi disease patients (mean, 0.02 logMAR in both eyes) but reduced in most RP patients (mean, 1.32 logMAR [right eye] and 1.35 logMAR [left eye]). Similarly, the visual field in the retinal area was preserved in Oguchi disease patients (mean, 677 mmRetinitis pigmentosa with SAG mutations often shows a characteristic golden sheen surrounding posterior pigmentary retinal degeneration. Oguchi disease can show progressive degeneration in adulthood, rarely resulting in RP.

Spinocerebellar ataxia type 1 (SCA1) caused by pathogenic CAG repeat expansion in the ATXN1 is characterized by loss of vision with little fundus abnormalities in some patients. Recently, macular degeneration has been reported to account for the visual symptoms in sporadic cases.Five consecutive patients diagnosed as SCA1 with supporting genetical evidence were newly referred to ophthalmology department from neurology unit. They underwent ocular examination to assess visual acuity and the structural integrity of the macula using optical coherent tomography (OCT). Full-field and multifocal electroretinogram (ERG) were recorded in some patients. Genetic testing was done by a polymerase chain reaction-based method.Fundus examinations revealed normal optic disc and macula appearance. However, four out of five patients had foveal thinning by OCT. This included three patients who showed reduced visual acuity. Among the three, multifocal ERG was performed in two, which showed reduced amplitudes in the localized foveal area. Full-field ERG showed normal responses in all five patients assessed. Only one patient had normal visual function and normal macular structure.Macular degeneration with subtle funduscopic alterations, sometimes mimicking occult macular dystrophy, is an important cause of visual loss in SCA1 patients, which could be reliably detected with OCT and multifocal ERGs.

Continuous drug administration with better adherence to treatment and less invasive procedures is important in treating retinal diseases such as age-related macular disease. In this study, we report a drug-refillable device consisting of a silicone reservoir and an injectable gelatin/chitosan gel (iGel). The silicone reservoir was fabricated with polydimethylsiloxane (PDMS) using a computer-aided design and manufacturing to have micropores at a releasing side for uniaxial release to the sclera. A stainless steel wire and sheet were combined in the side and bottom of the reservoir to ensure flexibility and to fit on the curvature of the eyeball and prevent irritation to the sclera through the bottom of the reservoir. The drug was injected and formulated in the reservoir by in situ crosslinking of gelatin/chitosan gel with the crosslinker; 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride. The in vitro release study using fluorescein molecules showed that the release rate from encapsulated iGel in the reservoir was slower than that from the original iGel. After reinjecting the iGel into the reservoir, the same release profile as the first injection was observed. The reservoir containing iGel was placed on the sclera of a rabbit and the distribution of 150 kDa fluorescein isothiocyanate-dextran (FD150) in the retina and choroid/retinal pigment epithelium (choroid/RPE) was studied. The cryosections showed that FD150 was observed in the choroid/RPE. Homogenates of the retina and choroid/RPE showed fluorescence during 12 weeks implantation, indicating the drug could be delivered to the retina by using the device. The drug filling was successful into the reservoir implanted on the sclera through the conjunctiva by using a needle. In conclusion, the refillable drug delivery device is a promising tool to administer drugs long-term by reinjection with less invasiveness to intraocular tissues.

Background/aimsTo investigate the relationship between aqueous flare, visual function and macular structures in retinitis pigmentosa (RP).MethodsClinical data from 123 patients with RP (227 eyes), 35 patients with macular dystrophy (68 eyes) and 148 controls (148 eyes) were analysed. The differences in aqueous flare between clinical entities and the correlation between aqueous flare (measured with a laser flare cell meter) versus visual acuity, visual field area (Goldmann perimetry) and macular thickness (optical coherence tomography) in patients with RP were determined. Influence of selected clinical data on flare was assessed using linear mixed-effects model.ResultsAqueous flare was higher in patients with RP than patients with macular dystrophy or controls (p=7.49×E−13). Aqueous flare was correlated with visual field area (R=−0.379, p=3.72×E−9), but not with visual acuity (R=0.083, p=0.215). Macular thickness (R=0.234, p=3.74×E−4), but not foveal thickness (R=0.122, p=0.067), was positively correlated with flare. Flare was not affected by the presence of macular complications. All these associations were maintained when the right and the left eyes were assessed separately. Analysis by linear mixed-effects model revealed that age (p=8.58×E-5), visual field area (p=8.01×E-7) and average macular thickness (p=0.037) were correlated with flare.ConclusionAqueous flare and visual field area were correlated in patients with RP. Aqueous flare may reflect the degree of overall retinal degeneration more closely than the local foveal impairment.

The disaster at the Fukushima Daiichi Nuclear Power Plant (FDNPP) remains unresolved because the estimated time to decommission a nuclear reactor appears to be approximately 40 years. The number of workers exposed to radiation doses ranging from 1 to 100 mSv continues to increase. To understand the accident progression at Fukushima and to anticipate what we should do in the future for occupational and environmental health, we performed a survey of citizens and doctors who lived inside and outside Fukushima in 2011 and 2013. In a comparison of these 2 years, the citizens inside Fukushima continue to suffer anxiety, although those living outside Fukushima tended to feel less anxious. Medical students who had recently studied radiation biology showed much less ongoing anxiety compared with other groups, suggesting that learning about the effects of radiation is essential to understanding one's own circumstances objectively and correctly. The lack of trust in the government and in the Tokyo Electric Power Company (TEPCO) in 2013 remains high in all groups. Therefore, long-term forthright explanations from the government, TEPCO, and radiation experts are indispensable not only to establish trust with people but also to alleviate psychological stress.

Successful treatment of age-related macular diseases requires an effective controlled drug release system with less invasive route of administration in the eye to reduce the burden of frequent intravitreal injections for patients. In this study, we developed an episcleral implantable device for sustained release of ranibizumab, and evaluated its efficacy on suppression of laser-induced choroidal neovascularization (CNV) in rats. We tested both biodegradable and non-biodegradable sheet-type devices consisting of crosslinked gelatin/chitosan (Gel/CS) and photopolymerized poly(ethyleneglycol) dimethacrylate that incorporated collagen microparticles (PEGDM/COL). In vitro release studies of FITC-labeled albumin showed a constant release from PEGDM/COL sheets compared to Gel/CS sheets. The Gel/CS sheets gradually biodegraded in the sclera during the 24-week implantation; however, the PEGDM/COL sheets did not degrade. FITC-albumin was detected in the retina during 18 weeks implantation in the PEGDM/COL sheet-treated group, and was detected in the Gel/CS sheet-treated group during 6 weeks implantation. CNV was suppressed 18 weeks after application of ranibizumab-loaded PEGDM/COL sheets compared to a placebo PEGDM/COL sheet-treated group, and to the intravitreal ranibizumab-injected group. In conclusion, the PEGDM/COL sheet device suppressed CNV via a transscleral administration route for 18 weeks, indicating that prolonged sustained ranibizumab release could reduce the burden of repeated intravitreal injections.

This report describes a rare case of a patient with growth hormone (GH)-secreting pituitary adenoma with malignant transformation resulting in multiple metastases to the dura mater of the cerebral convexity and high cervical spine. The patient was a 60-year-old man with a previous history of pituitary adenoma with suprasellar extension who had undergone transsphenoidal surgery, craniotomy for a convexity tumor, and suboccipital craniotomy for a cerebellar tumor. Thirteen years after the initial surgery, suboccipital craniotomy for a cervicomedullary junction tumor and cervicospinal surgery for a metastatic tumor was performed. Histologic findings of resected specimens demonstrated that the primary pituitary tumor was typical adenoma (similar to specimens from the initial surgery) but that the cerebellar and the dural tumor from the high cervical spine had a high incidence of mitotic figures, and cellular anaplasia with nuclear polymorphism and necrosis. In addition, the serum levels of GH were noted to have decreased with recurrence of the tumor. It was concluded that patients with pituitary adenoma, even when benign, must be carefully followed for signs of malignant transformation, and spinal or distant metastases.

BACKGROUND AND PURPOSE: Whether to treat UIAs is controversial. The aim of the study was to compare the clinical outcome of patients with UIAs who were either treated conservatively or preventively. MATERIALS AND METHODS: Patients with UIAs referred to our institution were prospectively enrolled in the study. Data collected included baseline characteristics, aneurysmal features, and procedural and follow-up information. Preventive treatment was recommended if the aneurysm was larger than 5 mm and was considered safely treatable. Endovascular surgery was the first-line therapy if the aneurysmal shape was appropriate for coiling. RESULTS: From January 2003 through April 2008, a total of 879 patients with 1110 UIAs were enrolled; 325 patients with 369 UIAs (mean size, 7.8 mm) were treated (treatment group), and 603 patients with 741 UIAs (mean size, 4.4 mm) were managed conservatively (observation group). Mean follow-up was 692.5 days (1405.5 person-years). In the observation group, 26 aneurysms (3.5%) had ruptured (1.8% per year; 1405.5 person-years), 10 patients died, and 7 were disabled (mRS, 3–6: 2.8%). Aneurysmal size was a significant risk factor for rupture (P = .001). The treatment group included aneurysms treated either with coiling (n=315), clipping (n=32), or a combined approach (n=9); 1 patient died, and 3 were disabled (mRS, 3–6: 1.2%). Therapeutic intervention was equal (UIAs of all sizes) or superior (UIAs > 5 mm; P = .025) to conservative management. CONCLUSIONS: Treatment of UIAs was justified in aneurysms larger than 5 mm, and EVS can be safely applied to nearly 90% of UIAs.

Subcutaneous space is a potential site for the transplantation of cells such as islets for treatment of type 1 diabetes. To enhance engraftment, an optimal space for the growth of the transplanted cells is needed along with neovascularization. In this study, we developed a device using a photocurable resin, poly(ethyleneglycol) dimethacrylates (PEGDM), for controlled release of basic fibroblast growth factor (bFGF) to create a subcutaneous neovascular bed in rats. The device consists of a disk-shaped capsule with micropores and is composed of tri(ethyleneglycol) dimethacrylate (TEGDM) and a drug formulation of PEGDM. The release rate was tuned by changing the number of pores and the composition of water and PEGDM in the drug formulation. bFGF released from devices incubated in phosphate-buffered saline (PBS) enhanced the growth of fibroblasts, indicating bioactivity of bFGF after release. Histological evaluation showed a significant increase in the extent of vasculature that was dependent on the amount of bFGF loaded into the device. A perfusion study using fluorescein isothiocyanate dextran 2000 kDa showed linear and capillary staining patterns, indicating potent functional vasculature. In conclusion, the controlled bFGF releasing device could provide a neovascular bed with the required vascularization in the subcutaneous space. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3017-3024, 2017.

We describe an electrochemical method of harvesting cells cultured on a biodegradable polymeric nanosheet (cell/nanosheet construct), which is stabilized on a self-assembled monolayer (SAM) of thiol molecules. A poly(lactic-co-glycolic acid) (PLGA) nanosheet was attached by hydrophobic interactions onto the surface of a SAM of l-cysteine coated onto a gold electrode. Retinal pigment epithelial cell lines (RPE-J cells) were cultured on the nanosheet to form a monolayer. An AA-size dry battery was used to apply a negative electrical potential, causing reductive desorption of the SAM from the gold surface. Within one minute of application of the voltage, the cell/nanosheet of several mm in diameter was successfully detached without the loss of cell viability in a gentle stream of the electrolyte solution. The use of a porous electrode shortened the detachment time due to the more efficient permeation of the electrolyte solution to the electrode surface. Cell transplantation following the harvesting process was demonstrated by the local delivery of RPE-J cell/nanosheet constructs into the subretinal space of rat eyes through a capillary needle. This nanosheet-based approach that allows the on-demand harvesting of cell/nanosheet constructs and their subsequent transplantation in a minimally-invasive manner could play an important role in cell transplantation therapy.

PURPOSE: To evaluate the efficacy of combined 25-gauge microincision vitrectomy surgery (MIVS) and toric intraocular lens (IOL) implantation with posterior capsulotomy. METHODS: Noncomparative, interventional case series performed at a single center. Twelve patients with vitreoretinal disease and cataracts, with preexisting regular corneal astigmatism greater than 1 diopter, underwent 25-gauge MIVS and toric IOL implantation with posterior capsulotomy. RESULTS: The toric IOL was successfully implanted in each case. At 6 months postoperatively, mean axis rotation was 5.7° ± 3.1°. At 1 month postoperatively, mean uncorrected and best corrected visual acuity improved; the improvement was maintained after 6 months. The absolute residual refractive cylinder was significantly lower postoperatively than the pre-existing regular corneal cylinder ( P = .003). There were no surgical complications except temporary posterior iris synechiae in one case. CONCLUSIONS: Combined 25-gauge MIVS and toric IOL implantation with posterior capsulotomy is a practical and safe method to treat vitreoretinal disease and cataracts with pre-existing corneal astigmatism. [ Ophthalmic Surg Lasers Imaging Retina . 2013;44:145–154]

Objective We report our initial experience of real-time intraoperative C-arm computed tomography (C-arm CT: DynaCT)–guided navigation surgery for pituitary tumors. Methods Recent advancement in flat panel technology makes it possible to obtain CT-like images by using rotation of the C-arm of a digital subtraction angiography (DSA) system. A specially designed new suite, which has C-arm CT-imaging-capable DSA in combination with a navigation system (VectorVision Sky, BrainLAB AG, Munich, Germany), allows neurosurgeons to perform endoscopic transsphenoidal procedures under real-time navigation support. Thirty-one pituitary tumor patients were examined. During or after tumor removal, contrast-enhanced DynaCT was conducted to rule out residual tumor in 12 cases. When enhanced tumor was confirmed, additional removal was continued without moving the patients. Results DynaCT and subsequent image transfer to navigation system was performed in all cases without difficulties, requiring only an additional 15 minutes of surgical time. Sellar fenestration in relation to tumors and absence of hidden hematomas was confirmed in all cases. The contrast-enhanced DynaCT was found to be contributing to a better handling of the residual tumor. In 9 of these 12 cases (75%), residual tumor was detected on DynaCT; consequently, further removal was considered. In 2 cases (16%) there was no enhanced lesion, indicating complete removal. Conclusions The proposed technique of intraoperative visualization in the hybrid operating room can be easy to perform and may be a useful adjunct to conventional transsphenoidal surgery for an improved resection rate and less cavernous sinus and internal carotid artery injury.

Background This study evaluates whether thermoreversible gelation polymer (TGP) can be used as a delivery device to deploy dermal fibroblasts and cytokines into experimental aneurysms in rats. Methods The right common iliac artery of rats was surgically ligated and an experimental aneurysm was created by applying exogenous elastase. Seven days later, two aneurysms were harvested and used as controls (Group A), two were embolized with pure TGP (Group B), two were embolized with TGP and basic fibroblast growth factor (bFGF) (Group C) and two were embolized with TGP loaded with rat dermal fibroblasts (Group D). The aneurysms were also embolized with TGP mixed with dermal fibroblasts and bFGF at different concentrations (10 ng/ml: Group E (n=2), 100 ng/ml: Group F (n=2), 1000 ng/ml: Group G (n=2)). Each aneurysm sample was harvested after 7 days and histologic analyses were performed. Results The most advanced thrombus organization in the aneurysm, such as prominent fibroblast proliferation and collagen deposition, was observed in Groups E, F and G, although there was no noticeable difference between the groups. Moderate thrombus organization was seen in Group D and minimal thrombus organization was seen in Groups B and C. Conclusions TGP mixed with both dermal fibroblasts and bFGF induced the most advanced thrombus organization in the experimental aneurysms followed by TGP mixed only with dermal fibroblasts. TGP may be useful as a delivery device to deploy fibroblasts and cytokines into aneurysms.

Photoreceptor apoptosis is a major cause of vision loss in many ocular diseases. Significant progress has been made to elucidate the molecular pathways involved in this process, yet little is known about proteins counteracting these apoptotic pathways. It is established that heat shock proteins (HSPs) function as molecular helper proteins (chaperones) by preventing protein aggregation and facilitating refolding of dysfunctional proteins, critical to the survival of all organisms. Here, we investigated the role of HSP70 on photoreceptor survival after experimental retinal detachment (RD) in mice and rats. We found that HSP70 was up-regulated after RD and associated with phosphorylated Akt, thereby preventing its dephosphorylation and further activation of cell death pathways. Administration of quercetin, which inhibits HSP70 and suppresses Akt phosphorylation significantly increased photoreceptor apoptosis. Similarly, RD-induced photoreceptor apoptosis was augmented in mice carrying hypomorphic mutations of the genes encoding HSP70. On the other hand, administration of geranylgeranylacetone, which induces an increase in HSP70 significantly decreased photoreceptor apoptosis after RD through prolonged activation of Akt pathway. Thus, HSP70 may be a favorable potential target to increase photoreceptor cell survival after RD.

A 34-year-old male presented with a rare case of Rathke's cleft cyst (RCC) with calcification manifesting as persistent high fever and impaired consciousness. Physical findings revealed panhypopituitarism and bitemporal hemianopsia. Computed tomography showed mass lesions with marked calcification within the sella turcica and the suprasellar region. Magnetic resonance imaging showed solid and cystic components compressing the optic nerve. The preoperative diagnosis was craniopharyngioma. Initial endonasal transsphenoidal surgery (TSS) was performed with a surgical microscope, but the mass was extremely hard, so only partial removal was possible. Second endonasal extended TSS was performed with a neuroendoscope. The solid components were totally removed, but calcifications adhering to the optic nerve could not be removed completely. The histological diagnosis was RCC with marked granulation reaction. RCC with calcification is rare and difficult to differentiate from craniopharyngioma on neuroimages. Extremely thick calcification of the sella turcica enclosing granulation tissue and the cyst similar to armor, here called "armor-like calcification," is a characteristic imaging finding of RCC with calcification. The most important aspect is choosing a surgical approach to carefully and effectively relieve pressure upon the optic nerve. Endonasal extended TSS with an endoscope was effective in the present case.

Fixation using an anterior cervical plate is frequently used in anterior cervical discectomy or corpectomy and fusion procedures because of the higher fusion rate than without instrumentation. The surgical outcomes with the anterior plate technique are acceptable, but various hardware-related complications have been reported. The authors describe a new surgical technique, called the kusabi fixation technique, for securing the bone strut during anterior cervical corpectomy using cylindrical cages. Following corpectomy, the trimmed bone strut was placed into the space drilled-out. Two small holes of 5-6-mm diameter and 5-mm depth were drilled out at the interface of two bones at the top and bottom of the bone strut in a diagonal orientation. Two cylindrical cages filled with autologous bone tips were tapped into the interface. Eight patients with myelopathy were treated by this method. All procedures were uneventfully performed as a single level surgery (two vertebral bodies and one disk level). Solid bone union was obtained in all patients at 6 months after the operation. Apparent alignment change in the cervical spine was observed in only one patient, who developed asymptomatic kyphosis. No instrumentation failure or significant complications related to the surgery occurred. Precise biomechanical aspects and long-term radiographic analysis compared with the plate fixation technique should be established, but this new method provides another way to secure the bone strut in anterior cervical corpectomy and fusion.

To investigate whether pan-retinal photocoagulation (PRP) affects vitreous levels of cytokines and macular thickening in patients with high-risk proliferative diabetic retinopathy (PDR).Fourteen patients with bilateral high-risk PDR--which requires pars plana vitrectomy (PPV)--but without a history of retinal photocoagulation participated in this study. Before PPV, one eye received PRP, and the other eye did not. The concentrations of cytokines of vascular endothelial growth factor (VEGF), stromal derived factor-1 (SDF-1), interleukin-6 (IL-6) and regulated upon activation, normal T-cell expressed and secreted (RANTES) were measured in each vitreous sample obtained at PPV. Macular thickness obtained by optical coherence tomography was also monitored during clinical course.After the PRP, macular thickness in PRP-pretreated eye showed temporal increase. Vitreous levels of IL-6 and RANTES in PRP pre-treated eye were significantly higher than levels in control eyes (p = 0.013, p = 0.033). Although macular thickness in control was correlated to vitreous levels of VEGF and IL-6 (p = 0.022, p = 0.003), that in PRP-pretreated eye was closely correlated to IL-6 and RANTES (p = 0.002, p = 0.011). After the PPV, macular thickness in both eye groups improved, and there was no significant difference between both eye groups 3 months after the PPV.In patients with high-risk PDR, PRP cause temporal worsening of macular edema linked with pro-inflammatory cytokines of IL-6 and RANTES, but not with VEGF and SDF-1. Thus, PRP-induced macular edema was caused by inflammation, while visual prognosis after PPV was not influenced by pretreatment of PRP.

Backgrounds and Purpose— The authors evaluated the incidence of rupture of unruptured intracranial saccular aneurysm during observation. Methods— Between January 2003 and December 2006, a total of 419 patients with 529 unruptured intracranial saccular aneurysms were observed without treatment. The mean follow-up duration was 905.3 days. Aneurysm size was measured by 3-dimensional CT angiography. Clinical and 3-dimensional CT angiography follow-up were obtained every 6 months. Results— Nineteen aneurysms ruptured during observation resulting in a 1.4% rupture rate per year. A history of subarachnoid hemorrhage (hazard ratio, 7.3; 95% CI, 2.5 to 21.2), posterior circulation aneurysm (hazard ratio, 2.9; 95% CI, 1.1 to 8), and large size were significant independent predictors for aneurysm rupture. Conclusions— Size, history of subarachnoid hemorrhage, and posterior circulation aneurysms were significant risk factors for prediction of rupture of unruptured intracranial saccular aneurysms.

The high mortality of acute subdural haematoma (ASDH) is largely explained by its frequent association with primary brain damage consisting of contusion and brain swelling. However, the nature and causes of brain swelling after traumatic brain injury are multifactorial and poorly understood. The purpose of this study was to investigate the pathophysiology of brain swelling associated with ASDH in traumatic brain injury.We examined whether the thickness of the haematoma, parenchymal injury, or presence of a secondary insult had an effect on traumatic brain swelling. The variables that might affect the pathophysiology of ASDH were examined, including: (1) age and mechanism of injury, (2) neurological findings, (3) secondary insult and extracranial injuries, (4) pre-operative computed tomography (CT) scan results, and (5) outcome.A total of 212 patients were included in this study. On CT scan, 159 patients (75.0%) did not have brain swelling, 29 (13.7%) had hemispheric brain swelling, and 24 (11.3%) had diffuse brain swelling. Brain swelling associated with ASDH is caused by secondary insult in addition to parenchymal injury. In the present study, the outcome of ASDH associated with brain swelling was poor, even when treated with early surgical evacuation; the mortality rate of such patients was over 75%.Given our findings, it is possible that the poor outcome of ASDH patients depends not only on the characteristics of the haematoma itself, but also on the presence of additional cerebral parenchymal injury and secondary insult.

Introduction DynaCT is a clinical application protocol to create computed tomographic (CT)-like images allowing soft-tissue visualization acquired from an angiography system. A cone beam three-dimensional CT reconstruction is produced from the acquisition of two-dimensional projection images by rotating the c-arm with x-ray source and image receptor around the patient. The purpose of this study is to evaluate the clinical efficacy of DynaCT in the operating room. Methods DynaCT was performed in 100 patients undergoing cerebral or spinal interventional procedures in the new-concept operating room. Specially designed AXIOM Artis BA (conventional image intensifier system; 55 patients; Siemens Medical Solutions, Erlangen, Germany) and AXIOM Artis dBA (flat-panel detector; 45 patients; Siemens Medical Solutions) biplane neuroangiographic systems (Siemens Medical Solutions, Germany) were installed in the operating room. The volumetric data set from the AXIOM Artis systems were reconstructed immediately on the three-dimensional workstation in the operating room. We compared DynaCT images with the use of multidetector computed tomography. Results DynaCT was performed successfully in all patients. High-contrast structures, such as bone, calcified lesions, and metallic materials, were visualized on DynaCT as well as in multidetector computed tomography for each group. Although contrast differentiation of soft tissue such as cerebral cortex, muscle, and hematoma on DynaCT were inferior to multidetector CT scans, the images were sufficiently used as intraoperative CT-like images. However, DynaCT images acquired from flat panel detector-based systems were found to be superior to those images acquired from image intensifier-based systems. Striking ring artifacts were exhibited and resulted in major limitations in the image intensifier group. Conclusion DynaCT has the potential to be used as a powerful tool for endovascular and neurosurgical procedures and will open new possibilities for neurosurgical management.

Background and Purpose— Transcranial ultrasound (TUS) enhances thrombolysis and is expected to be useful for the treatment of ischemic stroke. However, neither its effectiveness in improving neurologic outcome nor its safety in living tissue has been fully established. We examined the efficacy and safety of low-frequency TUS under appropriate conditions of ultrasound for thrombolytic treatment in a rat middle cerebral artery stroke model. Methods— Sixty-five male Wistar rats were used. Rats with right middle cerebral artery stroke exhibiting left hemiparesis were blindly selected and randomly assigned to 1 of 3 groups: (1) control, no therapy; (2) tPA, intravenous administration of tissue plasminogen activator 3 hours after middle cerebral artery stroke, or (3) TUS, tPA administration and application of TUS (490 kHz, continuous wave, at an intensity of 0.8 W/cm 2 ). Twenty-four hours after the onset of stroke, neurologic improvement was evaluated and brains were then removed. Thrombolysis at the origin of the right middle cerebral artery was examined. Thrombolysis ratio, cerebral infarct ratio, and rate of histologic evidence of hemorrhage were compared in the 3 groups. Results— Significantly better neurologic improvement ( P =0.008), a higher thrombolysis ratio ( P =0.041), and a reduction of cerebral infarct volume ( P =0.047) were obtained in the TUS group compared with the tPA group, without an increase in hemorrhagic transformation. Conclusions— Our findings suggest that thrombolytic treatment with low-frequency TUS under appropriate conditions could be an effective and safe method of treatment for ischemic stroke.

We evaluated the effects of a transscleral drug delivery device, consisting of a reservoir and controlled-release cover, which were made of photopolymerized polyethylene glycol dimethacrylate and triethylene glycol dimethacrylate, combined at different ratios. Geranylgeranylacetone (GGA), a heat-shock protein (HSP) inducer, was loaded into the device. The GGA was released from the device under zero-order kinetics. At both 1 week and 4 weeks after device implantation on rat sclera, HSP70 gene and protein expression were up-regulated in the sclera-choroid-retinal pigment epithelium fraction of rat eyes treated with the GGA-loaded device compared with rat eyes treated with saline-loaded devices or eyes of non-treated rats. Flash electroretinograms were recorded 4 days after white light exposure (8000 lx for 18 h). Electroretinographic amplitudes of the a- and b-waves were preserved significantly in rats treated with GGA-loaded devices compared with rats treated with saline-loaded devices. Histological examination showed that the outer nuclear layer thickness was preserved in rats that had the GGA-loaded device. These results may show that transscleral GGA delivery using our device may offer an alternative method to treat retinal diseases.

Objective and importance. Intracranial osteomas, which have no connections with the dura or the skull, are very rare. Here we report one case of multiple intracranial subarachnoid osteomas. Clinical description. A 24-year-old gentleman, who had no previous history of systemic disease, meningitis or head injury, presented with a 6-month history of non-specific intermittent headache over the whole head. Neuro-imaging showed multiple ossified lesions in the right frontal area, which were found to compress the underlying cerebral hemisphere. The patient underwent a right frontal craniotomy. The masses were dissected from the depressed brain along the intact pial planes, however, all of the white bone-hard masses tightly adhered to the superficial cortical veins. The masses were removed and the cortical veins were sacrificed. The patient had headaches and was nauseous for three days after the surgery due to venous congestion; however, the patient was neurologically intact on discharge. Pathological diagnosis was osteoma. Conclusion. To our knowledge, this is the first report of multiple intracranial subarachnoid osteomas. Surgical findings strongly support the hypothesis of the origin of the tumour that the primitive mesenchymal cells from the connective tissue might have migrated into the subarachnoid space accompanying the intracerebral blood vessels.

We analyzed the concentrations of four chemokines in the cerebrospinal fluid (CSF) and sera in Vogt–Koyanagi–Harada disease (VKH), an autoimmune uveomeningitis syndrome against melanocyte-associated proteins, with ELISA. CSF-CXCL10/IP-10 and CSF-CCL17/TARC were significantly elevated in VKH than in controls. In the majority of VKH cases and controls, CSF-CXCL10 was higher than serum-CXCL10, and CSF-CCL17 was lower than serum-CCL17. CCL11/Eotaxin was not different between groups. CSF-CCL2/MCP-1 was significantly lower in VKH than in control. The changes in VKH were essentially similar to those in multiple sclerosis, a known Th1-dominant condition.

Imaging features of intracranial haemangiopericytomas are similar to those of meningiomas. Preoperative identification of these tumours is important because of their aggressive nature, high rate of local recurrence and propensity for late metastasis. We reviewed the CT, MRI and angiographic findings in seven pathologically proved haemangiopericytomas, to determine if imaging characteristics might distinguish them from meningiomas. None showed hyperostosis or tumour calcification. All showed marked enhancement. Five had prominent internal signal voids, suggesting feeding arteries. On angiography, one had only pial-cortical supply but six also had meningeal supply; all showed a persistent stain. Only one had multiple "corkscrew" feeding vessels.

We report a case of branch retinal artery occlusion (BRAO) in a patient with iron-deficiency anemia. Various ophthalmological and laboratory studies were performed. A 32-year-old man had a sudden decrease of vision in his left eye to counting fingers at 30 cm two days ago. The left fundus showed a cherry-red spot and milky-white edema, except for the upper temporal region of the macula, and an optic disc malformation. Fluorescein angiography revealed leakage from the disc and a slightly delayed filling time in the left eye but an arterial filling defect was not noted. The differential diagnosis in this young patient includes polycythemia, hypercoagulopathy, coagulation abnormalities, trauma, hypertension, and autoimmune diseases such as systemic lupus erythematosus. Laboratory examinations revealed no abnormalities except for iron-deficiency anemia. The patient was treated with stellate ganglion block, hyperbaric oxygen, and ferrous sulfate. His visual acuity never recovered to better than 0.08. He had a coincidental rectal carcinoid and the tumor was excised surgically. No metastasis was observed. BRAO can be a complication of anemic retinopathy and can lead to severe visual loss without early medication.

Object. The authors have attempted to define the exact borders of the root exit zone (RExZ) of the facial nerve, measure the distribution of myelin histologically, and examine the relationship between contact vessels and the RExZ. Methods. Seventy-five facial nerves were obtained from brainstems excised from 44 adult patients at autopsy. The arteries and veins associated with the facial nerve were counted and measured. The facial nerves, associated vasculature, and adjoining portions of the brainstem were then removed en bloc. These tissues were serially sectioned and stained, and a photomicrograph of each section was obtained. The distribution of myelin on each section was measured from the upper edge of the supraolivary fossette, and the relationship between contact vessels and the RExZ was examined. The lateral transitional zone of the facial nerve began 8 mm distal to the upper edge of the supraolivary fossette (root exit point [RExP]) and had a mean length of 1.9 mm. The root detachment point (RDP) of the facial nerve at the medial side was located very close to the beginning of the medial transitional zone. In more than 80% of the nerves that were examined, vascular structures compressed the central glial myelin of the nerve. Conclusions. The authors propose the use of the terms “RExP,” “RDP,” and “transitional zone,” instead of RExZ, which cannot be well defined. The RDP appears to be a good landmark for use during microvascular decompression.

OBJECTIVE We have developed a modified laminoplasty procedure that preserves the posterior cervical elements, and we have used it to treat multilevel cervical canal stenosis, with or without spondylotic changes, in 30 patients. METHODS In this procedure, the posterior bony elements, such as the spinous processes, are not completely detached from the ligaments and musculature and are repositioned in the midline, to maintain normal cervical anatomic relationships. Thirty patients (mean age, 55.2 yr) were registered and evaluated in this study, with a mean follow-up period of 18.1 months, between November 1995 and October 2000. The rates of bony union of the reconstructed laminae and alignment of the cervical spine were assessed by using computed tomographic scans and lateral radiographs. RESULTS All patients except one noted some improvement in sensation, motor function, or both. Changes in spinal curvature were noted for two patients for whom a kyphotic deformity had developed before surgery (because of S-shaped or straight curvature). The average decrease in the range of motion was 7.26 degrees. Estimated rates of bony fusion at the sites of the gutters, spacers, and spinous processes were 94.6, 62.3, and 86.6%, respectively. A stable bony arch was assumed to be achieved with bony fusion at the gutters on the hinge side, as well as the spinous processes, approximately 6 months after surgery. CONCLUSION Our new technique of cervical laminoplasty preserves the posterior musculature and bony elements. This new procedure also helps maintain correct spinal alignment.

Cholesterol granuloma in the petrous apex presents with various symptoms of cranial nerve dysfunction, so the selection for surgical treatment remains controversial. We report a 41-year-old woman with a cholesterol granuloma at the left petrous apex, which was totally resected via a combined middle fossa and posterior transpetrosal approach. In a review of 92 cases, including our case, treated for petrous apex cholesterol granuloma between 1990 and 2001, 38 were men (41.3%) and 54 were women (58.7%). The mean age of these patients was 37.4 years (males were 35.2 years, females were 39.4 years). The most common presenting clinical symptom was hearing loss due to dysfunction of cranial nerve VIII. Seventy-nine patients, including our case, underwent operation, and 13 patients were managed without surgery. The most frequently selected surgical approach was middle cranial fossa approach. Revision surgery (including the 2 cases who underwent revision twice and three times) was performed in 12 patients (15.2%). In 92 cases, all of those who underwent total removal have not shown re-accumulation of cyst contents. On the other hand, 11.4% of the patients with or without permanent drainage route proceeded by subtotal removal of the cyst wall needed revision surgery because of stenosis of the drainage route. Therefore we conclude that the most important treatment for the prevention of re-accumulation of cyst contents may be the extent of the cyst wall resection rather than the establishment of permanent drainage route.

Several reports of clinical trials of immunotherapy using dendritic cells have been published to date. In this study, we investigated the safety and clinical response of immunotherapy with fusions of dendritic and glioma cells for the treatment of patients with malignant glioma. Eight patients with malignant glioma, ranging in age from 4 to 63 years old, participated in this study. Dendritic cells were generated from peripheral blood. Cultured autologous glioma cells were established from surgical specimens in each case. Fusion cells of dendritic and glioma cells were prepared with polyethylene glycol, and the fusion efficiency ranged from 9.2 to 35.3% (mean, 21.9%). All patients received the fusion cells every three weeks for a minimum of 3, and a maximum of 7, immunizations. Fusion cells were injected intradermally, close to a cervical lymph node. The percentage of CD16- and CD56-positive cells in peripheral blood lymphocytes slightly increased after immunization in 4 out of 5 cases investigated. Peripheral blood mononuclear cells were incubated with irradiated autologous glioma or U87MG cells and supernatants were harvested. In 6 cases analyzed, the concentration of interferon-gamma in the supernatant increased after immunization. Clinical results showed that there were no serious adverse effects and two partial responses. Although the results of the phase I clinical trial of fusion cells indicated that this treatment safely induced immune responses. we were unable to establish a statistically significant treatment-associated response rate, due to the limited sample population. Therefore, further evaluation of the role of adjuvant cytokines is necessary.

OBJECTIVE We used complementary deoxyribonucleic acid expression microarrays to assess the effects of radiotherapy on gene expression in glioblastoma multiforme. We hypothesized that postradiation recurrent tumors may demonstrate alterations in gene expression from the primary tumor specimen. METHODS Patients were diagnosed with glioblastoma multiforme at resection of the initial tumor, and they received 60 Gy of fractionated radiotherapy before recurrence. Ribonucleic acid samples from both the primary and the postradiation recurrent tumor in each patient were screened and compared using complementary deoxyribonucleic acid expression arrays and Northern blot analysis. RESULTS Messenger ribonucleic acid levels of growth factors participating in paracrine loops, such as vascular endothelial growth factor and platelet-derived growth factor receptor β, were decreased in postradiation recurrent tumors as compared with primary tumors in three of four patients. However, messenger ribonucleic acid levels of growth factors involved in autocrine loops, such as epidermal growth factor receptor, platelet-derived growth factor α, platelet-derived growth factor A, and basic fibroblast growth factor, were decreased in two of four, two of four, three of four, and three of four patients' recurrent tumors, respectively. Microvessel counts demonstrated that blood vessel growth was decreased significantly in postradiation recurrent tumor specimens. CONCLUSION After radiotherapy of glioblastoma multiforme, levels of paracrine-acting growth factors are diminished in correspondence with the reduction in vascular density. In contrast, growth factors that participate in autocrine loops demonstrate elevated levels of gene expression. These results suggest that maintenance of autocrine loops may be important in tumor regrowth after radiotherapy.

Wilson disease (WND) is caused by a deficiency of the copper-transporting enzyme, P-type ATPase (ATP7B). Twelve different mutations have previously been identified in Taiwan Chinese with Wilson disease. We, herein, report another 4 missense mutations, 1 of which is novel. We did haplotype analysis of Taiwanese WND chromosomes, using three well characterized short tandem repeat markers (haplotype was assigned in the order of D13S314-D13S301-D13S316). Association correlation was found between the mutations and their respective haplotypes. Haplotype-deduced pedigree analysis was shown to be helpful in the mutation analysis of WND chromosomes and in the molecular assessment of both pre-symptomatic WND patients and carriers. Given the complexity and heterogeneity of the mutation spectrum of ATP7B, we suggest that haplotype analysis should be performed before full-scale mutation analysis.

Purpose: A non-pigmented adenoma of the ciliary epithelium is rare. No good documentation has been reported in which magnetic resonance imaging (MRI), histological examination, and immunohistological evaluation have all been conducted. We showed how this tumor differed from ciliary body melanoma. Methods: We evaluated the ciliary body tumor by MRI, histological examination, and immunohistochemical examination. The data was compared with previously reported cases. Result: The tumor was classified as an adenoma of the non-pigmented ciliary epithelium with extracellular pools of hyaluronidase-sensitive mucopolysaccharide. The MRI appearance of the vitreous in our patient showed hyperintensity on T1, isointensity on T2 and marked enhancement with gadolinium. None of 3 other reported patients exhibited the same signals. Conclusion: MRI appearance is most variable in adenomas and no microscopical diagnosis can be based on that. Our patient showed isointensity on T2 with marked enhancement by gadolinium when compared to malignant melanoma.

Although a large amount of cholesterol is known to be needed for brain maturation and differentiation, cholesterol metabolism during these periods remains unclear. To elucidate the developmental regulation of cholesterol metabolism in the brain, we investigated the expression of 3-hydroxy-3-methyglutaryl-coenzyme A (HMG-CoA) reductase (EC 1.1.1.34), low-density-lipoprotein (LDL) receptor and very-low-density-lipoprotein (VLDL)/apolipoprotein E (apo E) receptor (VLDL receptor) using RNase protection assay (RPA) to quantitate mRNA levels in mouse brain, liver and kidney during development. Messenger RNA levels of HMG-CoA reductase in the brain decreased with age, and those levels at -5 (5 days before birth) and 5 days after birth were significantly higher than the control level of adult mice. The period from -5 to 5 days might correspond to stages of active biogenesis of the membranes of brain cells. The mRNA level of HMG-CoA reductase in the liver was also high at -5 days; a finding that correlated with cell proliferation. On the other hand, mRNA levels of the LDL and VLDL receptors in the brain did not change markedly during development. These results suggest that de novo cholesterol biosynthesis in brain cells plays a major role in the supply of cholesterol to the developing brain, rather than the uptake of cholesterol from serum lipoproteins through lipoprotein receptors.

Purpose: To explore whether the mutation in the retina-specific ATP-binding cassette transporter (ABCR) gene, the Stargardt's disease gene, contributes to the prevalence of the dry form of age-related macular degeneration (dry AMD) in Japanese unrelated patients. Methods: Twenty-five Japanese unrelated patients with dry AMD who were diagnosed by fluorescein angiography and indocyanine green angiography were chosen as the dry AMD group. None of these cases had apparent choroidal neovascularization. To detect the mutations in the ABCR gene, genomic DNA was extracted from leukocytes of peripheral blood, and 26 exons of the ABCR gene were amplified by polymerase chain reaction (PCR). All the PCR products were then directly sequenced. When a mutation was detected, the occurrence of a mutation was compared between these AMD patients and the control group. Results: After direct sequencing, a point mutation in exon 29 was found in one of the 25 dry AMD patients. In addition, a polymorphism in exon 45 was found in two other patients, and three sequence variations in exon 23 were detected in all patients. The incidence in AMD patients in whom a mutation in exon 29 (4%) was detected was less than that in controls (5%). Screening of the intron-exon boundaries also led to the identification of intronic mutation in intron 33. Conclusion: In this study we found no relationship between allelic variation in the ABCR gene and the prevalence of dry AMD in Japanese unrelated patients.

Purpose: To analyze the expression of metallothionein in neovascular membranes in patients with age-related macular degeneration (AMD) and to compare the findings in patients with proliferative diabetic retinopathy and proliferative vitreoretinopathy with findings in normal retinal pigment epithelial (RPE) cells. Methods: Semiquantitative reverse transcriptase-polymerase chain reaction and immunohistochemical examinations were performed. Sequence analysis was also carried out. Results: Expression of the metallothionein-II gene was not statistically significant in proliferative membranes. Sequence analysis revealed that there were at least six polymorphisms in the metallothionein-II gene and three in the metallothionein-IA gene. However, no amino acid substitution was observed. Metallothionein expression was also observed by immunohistochemical techniques in RPE cells of neovascular membranes in AMD patients. Conclusions: Metallothionein was reported to be decreased in aged macular RPE and in some monkey retina with early onset of macular degeneration. However, our results suggest that the expression of the metallothionein-II gene in neovascular membranes in patients with AMD may be controlled like other proliferating cells.

AIMS—Examination of the expression of the mutated allele of myotonic dystrophy protein kinase gene and lens epithelial cell changes in patients with myotonic dystrophy. METHODS—Six eyes from three patients with myotonic dystrophy underwent cataract surgery. The lens epithelium was photographed to examine the morphological changes. mRNAs were extracted to determine myotonic dystrophy protein kinase gene expression in the lens epithelium and peripheral blood. Age matched lens epithelial cells from senile cataracts were used as controls. RESULTS—All eyes showed iridescent or posterior subcapsular lens opacity. The expression of the myotonic dystrophy protein kinase gene with trinucleotide repeat expansion was evaluated by reverse transcriptase polymerase chain reaction, Southern blotting, and sequence analysis. Lens epithelial cell densities were extremely reduced in the patients compared with the control group. CONCLUSION—To the authors' knowledge, this is the first report to describe the relation between lens epithelial cell changes and mutated gene expression in patients with myotonic dystrophy. The gene may be mitotically unstable in the lens epithelial cells; it may influence cell density and lens epithelial function, and it may lead to the development of typical subcapsular lens opacity. Keywords: lens epithelial cell; myotonic dystrophy; RT-PCR

· Background: Herpes viruses may play an important role in acute retinal necrosis (ARN) syndrome, and raised antibody titer or virus genome have been detected in intraocular fluids of patients with ARN. In spite of aggressive anti-viral therapy, the clinical courses and final visual acuities among these patients have varied. Our purpose was to estimate the amount of virus and compare it to final visual acuity in patients with ARN. · Methods: The number of varicella-zoster virus copies in aqueous and vitreous before and after administration of anti-viral therapy were estimated by polymerase chain reaction (PCR) and by semi-nested PCR in 12 eyes of 11 patients with ARN and were compared with clinical characteristics. · Results: Viral genome DNA was amplified in ocular fluids diluted between 10−3 and 10−6 in patients whose final visual acuities were 0.3 or less. Conversely, the dilution required was between 10−1 to 10−3 in the patients whose final visual acuities were 0.4 or better. The difference was statistically significant. Further, patients in the latter group reacted to the anti-viral drug acyclovir more promptly than did the former patients. · Conclusion: Our results showed that final visual acuity may partly depend on the number of virus copies in ocular fluids. The number of virus copies tended to be higher in elderly or immunocompromised patients than in others, and the treatment was less efficacious in these patients.

We have shown previously in the spontaneously hypertensive rat (SHR) kidney that interleukin-6 (IL-6) and tumor necrosis factor (TNF) mRNA levels were low under conditions of acute anaesthesia and surgical stress. The reasons for the suppression of IL-6 and TNF gene expression in the SHR were investigated by examining the influence of enhanced beta-adrenergic stimulation, high blood pressure, and renal function (renal blood flow, glomerular filtration rate, plasma creatinine levels) on renal IL-6 and TNF mRNAs. The experiments were performed by means of the following three studies; (1) SHR and Wistar rats at 4, 7, 9 week old were injected with lipopolysaccaride (LPS), and then a relationship between blood pressure levels and IL-6 and TNF mRNA levels were estimated, (2) isoproterenol and propranolol were administered into SHR and WKY rats, and the levels of IL-6 and TNF mRNA were compared, (3) under condition of anaesthesia and surgical stress, blood pressure and renal functions in SHR were measured, and then the relationships between these factors and IL-6 or TNF mRNA levels were analyzed. Renal IL-6 and TNF mRNAs in SHR remained low even though blood pressure increased with age and there was no significant correlation between IL-6 or TNF mRNA levels and values of blood pressure or renal function under anaesthesia and surgical stress. However, the inhibition of the IL-6 and TNF mRNAs in SHR was prevented by propranolol treatment. These results suggested that suppression of IL-6 and TNF mRNAs in the SHR kidney could be due to overactivity of beta-adrenergic influences which may importantly contribute to the development of hypertension.

To examine the gene expression for melanogenesis of retinal pigment epithelial cells during dedifferentiation and to compare the condition to that of eyes obtaining anatomical success after surgery for proliferative vitreoretinopathy.Gene expression for melanogenesis was determined by reverse transcriptase-polymerase chain reaction of tyrosinase and tyrosinase-related protein-1 genes in normal and cultured retinal pigment epithelial cells and in proliferative membranes in patients with proliferative vitreoretinopathy.Gene expression for melanogenesis was classified into three types during dedifferentiation of retinal pigment epithelial cells: (1) tyrosinase-related protein-1 gene expression, (2) tyrosinase and tyrosinase-related protein-1 gene expression and (3) no expression of these genes. The expression of these genes were maintained better in mediums with basic fibroblast growth factor than in medium without basic fibroblast growth factor. Of the anatomically unsuccessful patients with proliferative vitreoretinopathy treated by surgery, 76.9% showed both tyrosinase and tyrosinase-related protein-1 gene expression; only 20% of the anatomically successful patients showed the gene expression.We reported three different conditions of retinal pigment epithelial cells based on gene expression for melanogenesis during dedifferentiation. The different condition of the retinal pigment epithelial cells may have some relationship to the anatomical results for proliferative vitreoretinopathy surgery.

We constructed brain-derived neurotrophic factor (BDNF) expressing rat retinal pigment epithelial (RPE) cells by stable transfection of BDNF cDNA, and the RPE cells were cultured on a cross-linked collagen sheet (Coll-RPE-BDNF). BDNF expression of the Coll-RPE-BDNF was confirmed by western blot, and the Coll-RPE-BDNF was transplanted into the rabbit sclera. In vivo BDNF expression was confirmed by His expression that was linked to the expressing BDNF. The effect of the released BDNF was examined in a rabbit acute high intraocular pressure system by electroretinogram and histological examination. Statistically significant preservation of ERG b wave amplitude was observed in the rabbits treated by Coll-RPE-BDNF when compared to that of no treatment. Statistically significant preservation of the thickness of the inner nuclear layer at the transplanted area was observed in the rabbits treated by Coll-RPE-BDNF compared to that of no treatment. Intra-scleral Coll-RPE-BDNF transplantation may partially rescue retinal cells from acute high intraocular pressure.

Triethylene tetramine dihydrochloride (trien 2HCl) has been used for the treatment of Wilson's disease, which is characterized by the accumulation of copper in various organs. We previously developed an HPLC system for analyzing trien, and found a trien metabolite in the urine when trien was orally given to humans. In this study, the metabolite was identified as 1-N-acetyltriethylene tetramine (acetyltrien) by FAB-MS and 1H-NMR spectroscopy. Trien and acetyltrien were capable of combining with copper, iron and zinc. However, the chelating activity of acetyltrien was significantly lower than that of trien. When trien was given to healthy adults, the amount of trien excreted in the urine was about 1% of the administered trien, whereas that of acetyltrien was about 8%. Most of the trien was excreted within the first 6 hours after the administration, while acetyltrien was excreted for over 26 hours. The urinary copper, iron and zinc levels all increased in parallel with the trien excretion.