Ultrafast spin-switching of a ferrimagnetic alloy at room temperature traced by resonant magneto-optical Kerr effect using a seeded free electron laser.

Ultrafast magnetization reversal of a ferrimagnetic metallic alloy GdFeCo was investigated by time-resolved resonant magneto-optical Kerr effect measurements using a seeded free electron laser. The GdFeCo alloy was pumped by a linearly polarized optical laser pulse, and the following temporal evolution of the magnetization of Fe in GdFeCo was element-selectively traced by a probe free electron laser pulse with a photon energy tuned to the Fe M-edge. The results have been measured using rotating analyzer ellipsometry method and confirmed magnetization switching caused by ultrafast heating.

Association study of genetic variants on chromosome 7q31 with susceptibility to normal tension glaucoma in a Japanese population.

The caveolin 1 to caveolin 2 (CAV1-CAV2) gene region on chromosome 7q31 has been reported to be associated with susceptibility to primary open angle glaucoma (POAG) and normal tension glaucoma (NTG) in previous studies. We investigated whether genetic variants in the CAV1-CAV2 region are associated with NTG in Japanese patients. Two hundred and ninety-two Japanese patients with NTG and 352 Japanese healthy controls were recruited. We genotyped three single-nucleotide polymorphisms; that is, rs1052990, rs4236601, and rs7795356, in the CAV1-CAV2 gene region and assessed the allelic diversity among cases and controls. The frequency of the minor allele (G) of rs1052990 was significantly decreased in NTG cases compared with controls (P=0.014, OR=0.71), whereas NTG or POAG cases had a significantly higher frequency of the allele than controls in previous studies. Conversely, rs7795356 did not show any significant association with NTG cases, and rs4236601 was monomorphic in the Japanese study population. Our findings did not correspond with previous positive results, suggesting that CAV1-CAV2 variants studied in the present study are not important risk factors for NTG susceptibility in all populations. Further studies are needed to elucidate the possible contribution of the CAV1-CAV2 region to the development of glaucoma.

Investigation of the association between the GLC3A locus and normal tension glaucoma in Japanese patients by microsatellite analysis.

PURPOSE: To investigate whether the GLC3A locus harboring the CYP1B1 gene is associated with normal tension glaucoma (NTG) in Japanese patients. MATERIALS AND METHODS: One hundred forty-two Japanese patients with NTG and 101 Japanese healthy controls were recruited. Patients exhibiting a comparatively early onset were selected as this suggests that genetic factors may show stronger involvement. Genotyping and assessment of allelic diversity was performed on 13 highly polymorphic microsatellite markers in and around the GLC3A locus. RESULTS: There were decreased frequencies of the 444 allele of D2S0416i and the 258 allele of D2S0425i in cases compared to controls (P = 0.022 and P = 0.034, respectively). However, this statistical significance disappeared when corrected (Pc > 0.05). We did not find any significant association between the remaining 11 microsatellite markers, including D2S177, which may be associated with CYP1B1, and NTG (P > 0.05). CONCLUSIONS: Our study showed no association between the GLCA3 locus and NTG, suggesting that the CYP1B1 gene, which is reportedly involved in a range of glaucoma phenotypes, may not be an associated factor in the pathogenesis of NTG.

Multi-layer amniotic membrane graft for pterygium in a patient with xeroderma pigmentosum.

BACKGROUND: Xeroderma pigmentosum (XP) is a rare, autosomal recessive, premalignant condition of the skin, and is reported to be associated with ocular surface disorders such as conjunctival malignancy and pterygium. Herein, we report a case of successful management of pterygium with multi-layer amniotic membrane graft (AMG) in a young XP patient. CASE: An 11-year-old Japanese girl, who had been diagnosed as having XP, was referred to us for treatment of her bilateral pterygium. Surgical intervention was attempted for a temporal, presumably fast-growing pterygium in her left eye. Multi-layer amniotic membrane grafting was performed. OBSERVATIONS: The surgery-induced pain and irritation disappeared within a day. The limbal conjunctival autograft survived on the AMG and re-epithelialization over the AMG was completed in 2 weeks. Best corrected visual acuity improved from 20/32 to 20/16 one month postoperatively. During the 1-year follow-up period, no recurrence was noted. CONCLUSIONS: The present case exemplifies that AMG as an adjunct to primary pterygium resection is effective even in a young patient with XP. In addition, multi-layer AMG, which we first demonstrated in this report, seems to be useful for protecting bare sclera and extraocular muscles from mechanical injury.

HRG4 (UNC119) mutation found in cone-rod dystrophy causes retinal degeneration in a transgenic model.

PURPOSE: To investigate the function and pathogenicity of HRG4, a photoreceptor synaptic protein homologous to the Caenorhabditis elegans neuroprotein UNC119. METHODS: HRG4 was screened for mutations in patients with various retinopathies, and a transgenic mouse model was constructed and analyzed based on a mutation found. RESULTS: A heterozygous premature termination codon mutation was found in a 57-year-old woman with late-onset cone-rod dystrophy. In some transgenic mice carrying the identical mutation, age-dependent fundus lesions developed accompanied by electroretinographic changes consistent with defects in photoreceptor synaptic transmission (depressed b-wave, normal c-wave), and retinal degeneration occurred with marked synaptic and possible transsynaptic degeneration. CONCLUSIONS: HRG4, the only synaptic protein known to be highly enriched in photoreceptor ribbon synapses, is now shown to be pathogenic when mutated.

Roles of the AGE-RAGE system in vascular injury in diabetes.

This study concerns whether advanced glycation endproducts (AGE) are related to microvascular derangement in diabetes, exemplified by pericyte loss and angiogenesis in retinopathy and by mesangial expansion in nephropathy. AGE caused a decrease in viable pericytes cultivated from bovine retina. On the other hand, AGE stimulated the growth and tube formation of human microvascular endothelial cells (EC), this being mediated by autocrine vascular endothelial growth factor. In AGE-exposed rat mesangial cells, type IV collagen synthesis was induced. Those AGE actions were dependent on a cell surface receptor for AGE (RAGE), because they were abolished by RAGE antisense or ribozyme. The AGE-RAGE system may thus participate in the development of diabetic microangiopathy. This proposition was supported by experiments with animal models; several indices characteristic of retinopathy were correlated with circulating AGE levels in OLETF rats. The predisposition to nephropathy was augmented in RAGE transgenic mice when they became diabetic.

Characterization of the gene for HRG4 (UNC119), a novel photoreceptor synaptic protein homologous to unc-119.

HRG4 (HGMW-approved symbol UNC119) is a novel human photoreceptor-enriched gene coding for a 240-amino-acid protein. Initially, HRG4 was shown to be 57% homologous to a newly discovered Caenorhabditis elegans gene, mutated in a coordination mutant and involved in chemosensation. Recently, HRG4 has been localized to the photoreceptor synapses in the outer plexiform layer of the retina. The HRG4 gene was cloned and characterized to facilitate its analysis as a potential pathogenic gene. The gene consisted of five coding exons, spread over approximately 8 kb of genomic DNA. The transcriptional start site was 14 bp upstream of the cDNA, 68 bp upstream of the putative translational initiation codon. Five GC boxes were identified in a 100-bp upstream region, along with a photoreceptor conserved element 1-like sequence at -603. Another photoreceptor gene-associated sequence, Ret-1, was present in intron 1, 71 bp downstream of the exon 1/intron 1 border. A CpG island encompassing exon 1 and sequences just before and after it was present. The gene was fine mapped to 17q11.2, facilitating its future consideration as a candidate for retinal diseases mapped to the same region.

Upregulation of retinal vascular endothelial growth factor mRNAs in spontaneously diabetic rats without ophthalmoscopic retinopathy. A possible participation of advanced glycation end products in the development of the early phase of diabetic retinopathy.

Vascular endothelial growth factor (VEGF) has recently been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. However, its involvement in the development of the early phase of diabetic retinopathy is not fully understood. In this study we investigated the retinal VEGF mRNA level in spontaneously diabetic Otsuka Long-Evans Tokushima fatty (OLETF) rats, a model of non-insulin-dependent diabetes, without overt retinopathy, using quantitative reverse-transcription polymerase chain reaction. The retinal VEGF mRNA level was 2.2 times higher (p < 0.0005) in OLETF rats than in control rats at the age of 60 weeks. Moreover, their retinal mRNA level was positively correlated with serum concentration of advanced glycation end products (AGEs) but not to serum glucose concentration. Furthermore, the peak latency of the oscillatory potentials in the electroretinogram, one of the most sensitive markers for the early phase of diabetic retinopathy, was significantly prolonged in OLETF rats (p < 0.05), being also correlated with the serum AGE concentration. The results thus suggest that AGEs, which are formed acceleratedly in diabetic conditions, are involved in the development of the early phase of diabetic retinopathy probably through the induction of retinal VEGF mRNAs.

Localization of HRG4, a photoreceptor protein homologous to Unc-119, in ribbon synapse.

PURPOSE: To characterize further HRG4, a novel photoreceptor protein recently identified by subtractive cDNA cloning, by sequence analysis and immunolocalization. METHODS: The rat homolog of HRG4, RRG4 was expressed and used to prepare an antibody. The antibody was used in Western blot analysis, and immunofluorescent localization at the light and electron microscopic levels of HRG4-RRG4 protein. The HRG4-RRG4 sequence was also analyzed for homologies. RESULTS: HRG4-RRG4 showed 57% homology with unc-119, a Caenorhabditis elegans neuroprotein causing defects in locomotion, feeding, and chemosensation when mutated. By Western blot analysis, the HRG4-RRG4 protein was demonstrable only in retina and was soluble in nature. Immunofluorescence microscopic study of human and rat retinas, using the HRG4-RRG4 antibody, and other rod and cone photoreceptor-specific antibodies showed that the HRG4-RRG4 protein is localized in the outer plexiform layer of the retina in the synaptic termini of rod and cone photoreceptors. Electron microscopic immunolocalization showed the protein in the cytoplasm and on the presynaptic membranes of the photoreceptor synapses. CONCLUSIONS: The homology to unc-119 and localization to the photoreceptor synapse are suggestive of a function for HRG4-RRG4 in photoreceptor neurotransmission. HRG4 is the first photoreceptor-enriched synaptic protein to be reported, suggesting that its function may be unique to the specialized ribbon synapses formed between photoreceptors and the horizontal and bipolar cells of the retina.

Effects of diurnal control in the mineral concentration of nutrient solution on tomato yield and nutrient absorption in hydroponics.

We researched effects of diurnal change of the mineral concentration on tomato yield and nutrient absorption. First, we examined the effect on yield in a spray culture, in the experiment 1-1, when nitrate concentration of solution (N) and potassium concentration (K) were low and phosphate concentration (P) was high during the daytime, while N and K were high and P was low during the night, the yield was low. In the experiment 1-2, when N and K were high and P was low during the daytime, while N and K were low and P was high during the night, the yield was low. Second, we examined the effect on nutrient absorption in a water culture. Concentration of KNO3, of solution was changed in the daytime or the night. When KNO3 level was low during the daytime, while it was high during the night, total nitrate and potassium absorption for 24 hours was the highest. It were showed the possibility of the efficient supply of minerals to plants by the diurnal control in minerals.

Immunolocalization of X-arrestin in human cone photoreceptors.

X-arrestin is a recently identified retina-specific gene of unknown function. Affinity-purified anti-peptide antibody to human X-arrestin was prepared, and used in Western blot analysis of human retinal proteins and for immunohistochemistry on human retinal sections. By Western blot analysis, the antibody specifically bound to an approximately 47 kDa protein, and by indirect immunofluorescence specifically labeled cone photoreceptors with greatest intensity in their outer segments. In single and double label experiments, the localization of X-arrestin immunoreactivity was compared with immunolabeling patterns obtained with antibodies to red/green cone opsin, rhodopsin, and S-antigen. The results showed that X-arrestin is expressed in red-, green- and blue-sensitive cones in the human retina.

Cloning of the cDNA for a novel photoreceptor protein.

A subtractive cDNA cloning strategy was used to isolate a 1381-base pair human retina-specific cDNA, human retinal gene 4 (HRG4), which hybridized to a 1.4-kilobase message in the retina and encoded a 240-amino acid acidic protein with a calculated molecular mass of 26,964 Da. The proximal 1/4 of the conceptual protein sequence was rich in glycine (18%) and proline (20%), had a predicted secondary structure of turns, and showed a loose similarity (19-24%) to various alpha-collagen sequences, while the distal 2/4 consisted of a mixture of alpha-helices, beta-sheets, and turns. Genomic Southern analysis with HRG4 showed cross-hybridizing sequences in six different species, and HRG4 was 92% homologous with a 1264-base pair rat cDNA (rat retinal gene 4; RRG4) at the protein level. The region of 100% identity between the two sequences corresponded to the distal 3/4 of the protein sequence consisting of mixed secondary structures, suggesting a functionally important domain. In vitro transcription and translation corroborated the open reading frames corresponding to HRG4 and RRG4 in the cDNAs. Expression of HRG4 in the retina was localized to the photoreceptors by in situ hybridization. Developmentally, RRG4 began to be highly expressed around postnatal day 5 in the rat outer retina when the photoreceptors begin to differentiate and rapidly increased in expression to reach the mature adult level by postnatal day 23. No diurnal fluctuation in expression of RRG4 was seen.

Effects of norfloxacin on the retina in rabbits.

BACKGROUND: Fluoroquinolones have a strong affinity with melanin, and their ocular effects have been reevaluated. Norfloxacin, one of the fluoroquinolones, has broad-spectrum activity against aerobic gram-positive and gram-negative bacteria. We examined the retinal toxicity and intraocular pharmacokinetics of intravitreal norfloxacin in rabbits. METHODS: Twenty-three albino and 23 pigmented rabbits were divided into three groups to evaluate retinal toxicity and two groups to investigate the intraocular pharmacokinetics. Each of these five groups was further divided into two subgroups (albino rabbits and pigmented rabbits). RESULTS: With 500 micrograms norfloxacin, the oscillatory potential of the electroretinogram was transiently and selectively deteriorated in albino and pigmented rabbits, whereas the electroretinogram remained unchanged with 50 micrograms in pigmented rabbits. No changes were observed in the visual evoked potential or on histology of the retina 7 days after an intravitreal injection of 50 or 500 micrograms norfloxacin. The electroretinogram and the retinal histology became abnormal 7 days after four intravitreal injections of 500 micrograms norfloxacin at 7-day intervals. As regards the intraocular pharmacokinetics after an intravitreal injection, the norfloxacin concentration in the chorioretina was as high as that in the vitreous 3 h after injection and was much higher than that in the vitreous 7 days after injection. Similar results were obtained after multiple injections. CONCLUSIONS: These results indicate a high concentration of norfloxacin in the melanin-containing ocular tissues.

[Endophthalmitis caused by Paecilomyces lilacinus after cataract surgery: a case report].

A case of fungal endophthalmitis caused by Paecilomyces lilacinus after cataract surgery is reported. An 84-year-old woman had been suffering from diabetes mellitus and hypertension. She underwent an uncomplicated extracapsular cataract extraction with implantation of a posterior chamber intraocular lens in the right eye. Approximately one month after the surgery iritis was noted, and treated systemically and topically with corticosteroids and antibiotics. The inflammation was improved by these therapies at first, but worsened when the dose of oral corticosteroid was reduced. White fluffy exudates were present on the iris near the chamber angle with the hypopyon. Removal of the intraocular lens, vitrectomy and medicinal therapy were undertaken. The eye was salvaged but the final visual acuity was counting fingers. Paecilomyces lilacinus was identified from the intraocular specimen taken at the time of surgery. The clinical effects and the results of in-vitro sensitivity test showed that fluconazole and amphotericin B were ineffective, and that econazole, thimerosal and miconazole were effective.

Sagittal magnetic resonance imaging of dysthyroid ophthalmopathy.

Sagittal magnetic resonance imaging (MRI) scans of the eye and orbit were made of 30 eyes in 15 cases of dysthyroid ophthalmopathy (DO). On the basis of these scans, we sought to elucidate relationships between the morphological condition of the levator palpebrae muscle, fatty tissue in the upper eyelid and the superior, inferior recti muscles and the occurrence of such symptoms as lid retraction, lid swelling and vertical disturbance of eye movement. The levator palpebrae muscle was enlarged in all 15 DO eyes (100%) with upper eyelid retraction. In 16 (88.9%) of 18 eyes with apparent lid swelling, enlargement of the preaponeurotic fat or submuscular fat pad was clearly evident. In the control eyes, no such enlargement was seen in either the levator palpebrae muscle or orbital fatty tissue. The clear space between the superior recti and the levator palpebrae muscles that was seen in control eyes was absent in all five eyes that presented a disturbance in infraduction. In 8 (80%) of 10 eyes with a disturbance in supraduction, the inferior rectus muscle was enlarged and muscle extension was impaired. Sagittal MRI seemed to be a useful means of obtaining a better clinical understanding of a variety of eye symptoms associated with DO.

Effect of intravitreal injection of norfloxacin on the retina in pigmented rabbits.

The effect of an intravitreal injection of norfloxacin on the retina was evaluated by in-vivo electroretinogram (ERG) and histological examination in pigmented rabbits. The intraocular pharmacokinetics after an intravitreal injection of norfloxacin were also investigated. An intravitreal injection of 50 micrograms norfloxacin produced no significant change in the ERG. An injection of 500 micrograms norfloxacin decreased the amplitude of the oscillatory potentials, and delayed their peak latencies 3 hours after the injection, but these changes recovered within 7 days. A marked suppression of the c-wave was noted in one pigmented rabbit. Neither 50 micrograms nor 500 micrograms norfloxacin caused any apparent changes in the visual evoked potential and in the retinal histology 7 days after the injection. The intraocular pharmacokinetic study showed that the concentration of norfloxacin in the choroid-retina was almost the same level as that in the vitreous body 3 hours after the injection. However, the former was higher than the latter 7 days after the injection, suggesting the persistency of norfloxacin in the choroid-retina. An intravitreal injection of 50 micrograms norfloxacin could be used without retinal toxicity. A high dose-intravitreal use of norfloxacin needs careful attention with respects to its persistency in the pigmented ocular tissues.

Correlated expression of mRNAs of carcinoembryonic antigen and nonspecific cross-reacting antigen genes in malignant and nonmalignant tissues of the colon.

Expression of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) mRNAs were observed in malignant and nonmalignant colon tissues by Northern blot analysis. CEA mRNA was detected together with NCA mRNA in nine cultured cell lines, with the exception of the lung carcinoma A549 cell line, and in 19 colon tissue specimens, including carcinomas, adjacent noninvaded tissues and adenomas. When we compared the intensity of the hybridization signal for CEA or NCA mRNA among adenomas, carcinomas and adjacent noninvaded tissues, NCA mRNA rather than CEA mRNA was highly expressed in carcinomas compared to adjacent nonivaded tissues. When, however, the relation between the intensities of hybridization signals for CEA and NCA mRNAs were evaluated in five colon carcinoma cell lines and 19 colon tissue specimens, a statistically significant correlation was observed (gamma = 0.462, P less than 0.01). These data suggest that NCA may be more useful as a tumor marker for colon carcinoma than CEA at the level of mRNA, and that transcriptions of the CEA and NCA genes might be regulated by some common mechanisms in malignant and nonmalignant tissues of the colon.

[Detection of the new tumor marker MUSE 11 antigen in sera of pancreatic cancer patients: a comparison with sialyl SSEA-I antigen].

Using sera from patients with pancreatic cancer and chronic pancreatitis, we measured the level of the adenocarcinoma-associated antigen MUSE 11. A comparative study between levels of MUSE 11 and levels of sialyl SSEA-1 antigen (SLX) was also carried out. With respect to the MUSE 11 antigen, positive incidence was found in 17 out of 26 pancreatic cancer patients (65%), and in 1 out of 13 chronic pancreatitis patients (8%). Similar results were obtained from the assay of SLX levels. However, no correlation was found between the two markers. Out of 9 samples which showed MUSE 11 negative, three were positive for SLX. Out of 13 samples showing SLX negative, seven were positive for MUSE 11. Twenty out of 26 (77%) cases showed positive results for either antigen. These data suggest that the MUSE 11 antigen may be useful for the diagnosis of pancreatic cancers.

Role of rectus muscle enlargement in clinical profile of dysthyroid ophthalmopathy.

Using logistic regression analysis, a correlation between enlargement patterns of the 4 rectus muscles and the occurrence of a variety of dysthyroid ophthalmopathy-associated ocular symptoms were analyzed in 698 eyes of 349 patients with this disorder. The condition of each rectus muscle was evaluated by computed tomography, and the various ocular symptoms were classified according to the Inoue classification system. The enlargement pattern, particularly when involving the inferior and medial recti, was found to have a strong statistical correlation with the occurrence of individual ocular symptoms. Additionally, the probabilities of the occurrence of most of the ocular symptoms were generally greater in those eyes with 2-, 3- or 4-muscle enlargement patterns than in those eyes with one or no enlarged rectus muscle.

Detection of mRNAs of carcinoembryonic antigen and nonspecific cross-reacting antigen genes in colorectal adenomas and carcinomas by in situ hybridization.

Expression of mRNAs of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) genes in colorectal carcinomas and adenomas was investigated by in situ hybridization (ISH) with specific biotinylated probes. Hybridization was clearly detected throughout the cytoplasm of 7 out of 15 adenomas and 13 out of 15 carcinomas with the CEA cDNA probe, and in 6 out of 15 adenomas and 10 out of 15 carcinomas with the NCA cDNA probe. The intensity of signal appeared to be stronger in carcinomas than that in adenomas, and the CEA and NCA mRNAs were expressed together in most of the positive tissue specimens. On the other hand, noninvaded tissues adjacent to the carcinoma did not show any signal except for 4 cases faintly stained with the NCA probe. This finding was partly confirmed by Northern blot analysis which indicated that the specific bands for the CEA and NCA mRNAs were more intense in RNAs from carcinoma tissues than in those from adjacent noninvaded tissues. These data suggest that the ISH technique with biotinylated probes could be of use for analyzing expression and localization of CEA and related genes on tissue sections.