Precise Measurement of Differential Cross Sections of the Σ

The differential cross sections of the Σ^{-}p→Λn reaction were measured accurately for the Σ^{-} momentum (p_{Σ}) ranging from 470 to 650 MeV/c at the J-PARC Hadron Experimental Facility. Precise angular information about the Σ^{-}p→Λn reaction was obtained for the first time by detecting approximately 100 reaction events at each angular step of Δcosθ=0.1. The obtained differential cross sections show a slightly forward-peaking structure in the measured momentum regions. The cross sections integrated for -0.7≤cosθ≤1.0 were obtained as 22.5±0.68 [statistical error(stat.)] ±0.65 [systematic error(syst.)] mb and 15.8±0.83(stat)±0.52(syst) mb for 470

Correction to: Comprehensive genetic analyses using targeted next-generation sequencing and genotype-phenotype correlations in 53 Japanese patients with osteogenesis imperfecta.

The original article has been corrected.

Comprehensive genetic analyses using targeted next-generation sequencing and genotype-phenotype correlations in 53 Japanese patients with osteogenesis imperfecta.

To elucidate mutation spectrum and genotype-phenotype correlations in Japanese patients with OI, we conducted comprehensive genetic analyses using NGS, as this had not been analyzed comprehensively in this patient population. Most mutations were located on COL1A1 and COL1A2. Glycine substitutions in COL1A1 resulted in the severe phenotype. INTRODUCTION: Most cases of osteogenesis imperfecta (OI) are caused by mutations in COL1A1 or COL1A2, which encode α chains of type I collagen. However, mutations in at least 16 other genes also cause OI. The mutation spectrum in Japanese patients with OI has not been comprehensively analyzed, as it is difficult to identify using classical Sanger sequencing. In this study, we aimed to reveal the mutation spectrum and genotype-phenotype correlations in Japanese patients with OI using next-generation sequencing (NGS). METHODS: We designed a capture panel for sequencing 15 candidate OI genes and 19 candidate genes that are associated with bone fragility or Wnt signaling. Using NGS, we examined 53 Japanese patients with OI from unrelated families. RESULTS: Pathogenic mutations were detected in 43 out of 53 individuals. All mutations were heterozygous. Among the 43 individuals, 40 variants were identified including 15 novel mutations. We found these mutations in COL1A1 (n = 30, 69.8%), COL1A2 (n = 12, 27.9%), and IFITM5 (n = 1, 2.3%). Patients with glycine substitution on COL1A1 had a higher frequency of fractures and were more severely short-statured. Although no significant genotype-phenotype correlation was observed for bone mineral density, the trabecular bone score was significantly lower in patients with glycine substitutions. CONCLUSION: We identified pathogenic mutations in 81% of our Japanese patients with OI. Most mutations were located on COL1A1 and COL1A2. This study revealed that glycine substitutions on COL1A1 resulted in the severe phenotype among Japanese patients with OI.

Marked efficacy of combined three-drug therapy (Sodium Valproate, Topiramate and Stiripentol) in a patient with Dravet syndrome.

WHAT IS KNOWN AND OBJECTIVE: Dravet syndrome (DS) is an intractable epilepsy syndrome. The three-drug combination therapy of sodium valproate (VPA), clobazam (CLB) and stiripentol (STP) is recommended worldwide. CASE SUMMARY: We present a case of DS, in which treatment with CLB could not be continued because of the appearance of adverse reactions to it. Replacement with topiramate (TPM) proved to be markedly effective. WHAT IS NEW AND CONCLUSION: It is suggested that combination therapy with VPA, TPM and STP is for DS epilepsy.

Ultracytochemistry of calmodulin binding sites in myocardial cells by staining of frozen thin sections with colloidal gold-labeled calmodulin.

Calmodulin (CaM) has been implicated as a multifunctional regulator of Ca2+ in the cytoplasm of cells. We have recently introduced biologically active colloidal gold-labeled CaM as a marker for identifying potential CaM binding sites (unoccupied by endogenous CaM at the time of fixation) by electron microscopy and have stained frozen thin sections of rat cardiac muscle with this conjugate. In the presence of Ca2+, gold particles indicating CaM binding sites were found localized on the sarcoplasmic reticulum, mitochondria, and gap junctions. Control tissue sections treated with EGTA or exposed to excess amounts of unlabeled native CaM before staining showed no binding. We believe that cytochemistry of potential CaM binding sites revealed by staining with labeled exogenous CaM is useful in correlating known biochemical reactions of CaM with particular cell activities.

Cytochemical studies on pathological Müller cells after argon laser photocoagulation.

The cytochemical localization of G6Pase activity, which is specific to the endoplasmic reticulum (ER) of Müller cells, was studied after argon laser photocoagulation in the guinea pig retina. After argon laser radiation, Müller cells exhibited enlargement of the cytoplasm, an increase of reactive ER and the nuclei, dislocation of the nuclei and diagonal stretching of the cytoplasm. However, cell attachment between Müller cells and the proliferated pigment epithelial cells or Bruch's membrane differed with the degree of retinal coagulation. This histo- and cytochemical method may be useful for examining Müller cells under various pathological conditions.

Distribution of FGF-5 in the rhesus macaque retina.

PURPOSE: To determine the distribution of mRNA transcribed from the FGF-5 gene and the distribution of FGF-5 protein in the normal adult retina of the rhesus macaque. METHODS: Freshly enucleated globes from rhesus macaque were prepared for Northern blot analysis, in situ hybridization, and immunohistochemical studies. A 350 base pair sequence from the human FGF-5 was subcloned and used to prepare 35S-labeled cRNAs for Northern and in situ experiments. Antipeptide antibodies were raised against an aminoterminal sequence as well as an internal sequence and were affinity purified on recombinant FGF-5 coupled to AffiGel-10. The specificity of antibody probes was verified by dot blot and Western blot analyses. RESULTS: Ganglion cells and photoreceptors express the highest levels of FGF-5 mRNA, although all neurons and the retinal pigment epithelium (RPE) were clearly labeled. Immunohistochemical staining similarly revealed a widespread distribution of the protein, with prominent labeling observed in photoreceptor inner segments, especially in cone cells. Plexiform layers and RPE cells were also clearly labeled. CONCLUSIONS: Clear evidence was established for the expression of FGF-5 in the retina, where it may play an important extracellular role as a secreted member of the FGF gene family.

Developmental expression of bFGF in the bovine retina.

PURPOSE: To examine the expression of bFGF in the developing bovine retina. METHODS: Fetal bovine eyes at 90, 120, 150, and 180 days gestational age, as well as adult bovine eyes, were immunohistochemically stained for the presence of bFGF. Detailed characterization of the anti-bFGF antibodies by immunoblot and Western blot analysis against pure FGF gene family standards and crude extracts of bovine retina were also performed. RESULTS: Expression of bFGF occurs beginning at 150 days of gestation, a period when photoreceptor development and secondary capillary network development is in process. No bFGF expression was found at 90 days, but primary capillaries were already apparent at this stage of development. CONCLUSIONS: Expression of bFGF in the developing bovine retina may play a functional role in outer retinal development.

Two Thai families with Norrie disease (ND): association of two novel missense mutations with severe ND phenotype, seizures, and a manifesting carrier.

We describe two Thai families with Norrie disease (ND) in three generations, including 10 affected males and one manifesting female. All affected males in each family had severely defective eye development with complete loss of vision. In addition, three male patients (one from family 1 and two from family 2) suffered from epilepsy, and one female carrier from one family manifested blindness with phthisis bulbi in her right eye. Mutation analysis of the ND gene (NDP) revealed two different novel missense mutations (L16P and S75P) that co-segregated with ND in each family, suggesting that the newly appearing proline at codon 16 or codon 75 alters the conformation of the ND protein and contributes to the severe phenotype of ND in each family. Other studies suggest that epileptic seizures or growth retardation that is associated with ND is the consequence of loss of contiguous genes, because most such patients had deletions extending beyond the Norrie locus. Our finding that the three affected males in the two families with the missense mutations had epilepsy does not support a contiguous gene effect, but favors the pleiotropism of NDP, at least as far as the epileptic manifestation is concerned. The unilateral blindness in the female carrier may have been due to non-random X-inactivation.

HTLV-I-associated myelopathy associated with multi-organ inflammatory disease: a case report.

We report a 73-year-old man with human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (HAM) complicated with multi-organ inflammatory disease, including Sjögren's syndrome, interstitial cystitis, and uveitis. The presence of HTLV-I proviral DNA in peripheral blood mononuclear cells (PBMC), cerebrospinal fluid, salivary gland, mucosa of urinary bladder, and aqueous humor was confirmed by polymerase chain reaction using HTLV-I pX region primer. Western blot analysis revealed the presence of anti-HTLV-I antibodies in serum, CSF, saliva, and urine, suggesting replication of HTLV-I in each tissue. A high load of HTLV-I proviral DNA (20 copies out of 100 PBMC) was present, associated with increased spontaneous proliferation of peripheral blood lymphocytes (24,747 cpm). Our results suggest that the high load of HTLV-I in patients with HAM may potentially induce systemic inflammation in several organs.

Conradi-Hünermann syndrome with ocular anomalies.

We report a Japanese girl with the Conradi-Hünermann form of chondrodysplasia punctata and anterior segment malformations characteristic of Axenfeld-Rieger syndrome. The patient also had cataracts and unilateral optic atrophy. A possible role for homeobox-containing genes in the etiology of this type of chondrodysplasia punctata is suggested as an explanation for the coincidence of these two syndromes.

Expression of the MAS proto-oncogene in the retinal pigment epithelium of the rhesus macaque.

The MAS proto-oncogene codes for a seven transmembrane protein which has been previously localized to specific regions of the hippocampus and cerebral cortex in the rat central nervous system. Because MAS has biological properties related to the growth and differentiation of cells of neuroectodermal origin, we investigated the distribution of MAS expression in the rhesus macaque retina by in situ hybridization. A 330 base pair (bp) segment of the human MAS sequence was subcloned and used to generate single-stranded cRNA probes for these studies. Our results demonstrated little, if any, positive signal over the neurons of the retina. The use of epi-polarization microscopy, however, revealed a distinct positive labelling of retinal pigment epithelial (RPE) cells. These studies suggest the use of MAS as a possible marker for the retinal pigment epithelium.

Increasing cell density down-regulates the expression of acidic FGF by human RPE cells in vitro.

Previous studies have reported the expression of acidic fibroblast growth factor (aFGF) by rat, bovine, and human retinal pigment epithelium (RPE) in vivo. To critically examine the expression of aFGF by RPE cells, we studied the density dependence of steady-state levels of mRNA and protein expression in vitro. Northern blot analysis demonstrated 5 transcripts ranging from 4.5 kB to 1 kB. Steady-state levels of all the transcripts decreased as a function of culture density. A polyclonal antibody was raised against recombinant human aFGF and affinity purified on aFGF coupled to AffiGel-10. The resulting antibody crossreacted with bFGF but not FGF-5, but this crossreactivity could be eliminated by absorption of the antibody on bFGF coupled to AffiGel-10. The final antibody preparation recognized only a single band at approximately 18.5 kD in lysates of RPE. Immunohistochemical staining with this antibody preparation demonstrated a marked dependence on cell density after 3 days in culture. Low culture density yielded cells staining moderately for aFGF, while confluent cells exhibited little or no staining. The reduction of aFGF from RPE cells in culture in a density-dependent fashion could also be demonstrated by Western blot analysis.

Expression of FGF5 in choroidal neovascular membranes associated with ARMD.

PURPOSE: The fibroblast growth factors (FGFs) are a family of 9 heparin binding proteins which have been proposed to play key roles in angiogenesis. Basic FGF (bFGF), acidic FGF (aFGF) and FGF5 have previously been demonstrated to be expressed in the normal retina and RPE. In this study, the expression of FGF5 was explored in choroidal neovascular membranes removed from patients with age-related macular degeneration (ARMD). METHOD: Three membranes were surgically removed from patients with ARMD, and were fixed, embedded and sectioned for immunohistochemistry. The membranes were immunostained with an affinity purified rabbit polyclonal antibody raised against the amino acid sequence for residues 175 to 185 of human FGF5 and visualized with the silver enhanced colloidal gold method for light microscopy. RESULTS: FGF5 was expressed in membranes arising from ARMD, and was found primarily in blood vessels and the surrounding extracellular matrix. CONCLUSIONS: These results suggest that FGF5 may have a functional role in the pathophysiology of ARMD.

Immunohistochemical study of p53, p21 and PCNA in pterygium.

Since mutated p53 is one of the most frequent gene abnormalities in human cancer, we hypothesized that mutation of p53 may play an important role in growth and recurrence of pterygia, a dysplasia of the conjunctiva. Therefore, we compared pterygia of Japanese and Tunisian patients using antibodies against p53, p21 and proliferating cell nuclear antigen (PCNA). In Nagasaki, 21 pterygia of Japanese individuals were removed and in Gabes, 19 primary pterygia of Tunisian individuals. Positive staining of wild type p53 was not found in the Japanese pterygia, whereas 38.1% were positive for mutant p53, none were positive for p21 and 76.2% were positive for PCNA. The incidence of mutant p53-positive staining was 50.0% in males and 22.2% in females, which was statistically significant. In the 19 Tunisian patients, positive staining of wild type p53 was not found, whereas 36.8% were positive for mutant p53, 0% for p21 and 63.1% for PCNA. Differences between Japanese patients and Tunisian patients were not significant. There were 2 types of pterygium. One type did not show mutant p53 and the other showed mutant p53 caused by ultraviolet light. However, damage caused by p53-dependent programmed cell death of pterygium cells may lead to mutations in other genes which may allow the progressive multistep development of limbal tumors. It is possible that mutant p53-positive pterygia can develop into limbal tumors.

Orbital lobe lacrimal ductal cyst.

Lacrimal ductal cysts are rare, especially in the orbit. A case of lacrimal ductal cyst in the orbital lobe, revealed by computed tomography and magnetic resonance imaging, is described in a 16-year-old man who was successfully treated by complete surgical excision. The radiologic and clinical features of this type of lacrimal ductal cyst are discussed.

Removal of indoor pollutants under UV irradiation by a composite TiO2-zeolite sheet prepared using a papermaking technique.

Toluene and formaldehyde are malodorous and cause indoor pollution. These materials have received much attention as hazardous and malodorous substances. It is well known that long-term exposure to even fairly low levels of toluene and formaldehyde brings about the risk of asthma and eczema. In this study, a composite TiO2-zeolite (ZE) sheet prepared using a papermaking technique was applied to remove toluene and formaldehyde under UV irradiation. The optimum composition of the TiO2 (Ti)-ZE sheet was studied in detail with regard to the effective removal of various indoor pollutants. Gaseous toluene and formaldehyde were removed by a composite TiO2-ZE sheet with different efficiencies depending upon the ratio of Ti/ZE in the composite sheet. The composite sheets could decompose formaldehyde and toluene repeatedly after being recharged. It was shown that the sheets are potentially applicable as highly functional materials to be placed on walls and ceilings of houses for the removal of various indoor pollutants.

Photocatalytic oxidation of NOx using composite sheets containing TiO2 and a metal compound.

The photocatalytic oxidation of nitrogen oxides (NO(x)) over titanium dioxide (TiO(2)) sheets containing metal compounds (MCs) had been studied. Calcium oxide (CaO), magnesium oxide (MgO), calcium carbonate (CaCO(3)), aluminium oxide (Al(2)O(3)) and ferric oxide (Fe(2)O(3)) were used as MCs. Al(2)O(3) and Fe(2)O(3) added to the TiO(2) sheet did not affect the photooxidation of nitrogen oxides (NO(x)). The CaO sheet treated with TiO(2) sol had the greatest efficiency as a NO(x) remover under UV irradiation. It is believed that CaO has a high adsorptivity for nitrogen dioxide (NO(2)) and nitric acid (HNO(3)). The amount of NO(x) removed by a TiO(2) sheet including MC showed a tendency to increase with increasing pH of the MC suspension, i.e. there is a good correlation between the alkalinity of the MC and the retention of NO(2) and HNO(3).

Histologic study of living response to artificially synthesized hydroxyapatite implant: 1-year follow-up.

We implanted artificially synthesized hydroxyapatite spheres into the orbits of 13 rabbits after enucleation. The spheres were removed 1, 2, 3, 6, and 12 months after implantation and examined by light and scanning electron microscopy. Tissue breakdown and exposure of the artificially synthesized hydroxyapatite implants were not observed. Month after month, fibrovascular tissues gradually invaded the pores of the artificially synthesized hydroxyapatite spheres deeper and deeper. Although the hydroxyapatite we used was completely artificially synthesized, we observed a mild foreign-body reaction around the artificially synthesized hydroxyapatite spheres. After 12 months, however, relief of the foreign-body reaction had occurred. Hydroxyapatite spheres for orbital implants after enucleation without scleral enveloping are appropriate.

Effect of delayed argatroban treatment on intracerebral hemorrhage-induced edema in the rat.

Studies indicate that thrombin plays an important role in intracerebral hemorrhage (ICH) induced edema formation. However, the time window for administration of a thrombin inhibitor to reduce ICH-induced edema is unknown. Nor is it known whether this time window extends beyond the period when a thrombin inhibitor might exacerbate rebleeding. This study examines whether a thrombin inhibitor, argatroban, can reduce edema formation following intracerebral infusion of 100 microl of blood in the rat, the therapeutic time window for argatroban, and whether argatroban promotes rebleeding. Intracerebral injection of argatroban 3 hours after ICH caused a significant reduction in edema measured at 48 hours. The systemic administration of argatroban (0.9 mg/h) starting 6 hours after ICH also significantly reduced edema formation. There was no protection when the onset of argatroban administration was delayed to 24 hours after ICH. Argatroban did not increase collagenase-induced hematoma volume when given into the clot after 3 hours or given systemically at 6 hours. Our data suggest argatroban may be an effective therapy for ICH-induced edema.