Efficacy and safety of retinol palmitate ophthalmic solution in the treatment of dry eye: a Japanese Phase II clinical trial.

PURPOSE: The purpose of this study was to investigate the efficacy and safety of the administration of retinol palmitate (VApal) ophthalmic solution (500 IU/mL) for the treatment of patients with dry eye. PATIENTS AND METHODS: This study included 66 patients with dry eye. After a 2-week washout period, patients were randomized (1:1) into either a VApal ophthalmic solution or a placebo group, and a single drop of either solution was administered six times daily for 4 weeks. Efficacy measures were 12 subjective symptoms, rose bengal (RB) and fluorescein staining scores, tear film breakup time, and tear secretion. Safety measures included clinical blood and urine analyses and adverse event recordings. RESULTS: In comparisons of the two groups, the mean change in RB staining score from baseline was significantly lower in the VApal group at 2 and 4 weeks (P<0.05 and P<0.01, respectively). Furthermore, the fluorescein clearance rate (fluorescein staining score) was significantly higher in the VApal group at 4 weeks (P<0.05). The VApal group showed a significant improvement in blurred vision at 1 and 2 weeks (P<0.01 and P<0.05, respectively), and the mean change in the total score for subjective symptoms from baseline was significantly lower in the VApal group at 1 week (P<0.05). In before- and after-intervention comparisons, the fluorescein and RB staining scores showed improvement in both groups. Improvement was noted for 11 subjective symptoms in the VApal group and for seven symptoms in the placebo group. No significant differences in adverse events and reactions were found between the groups. CONCLUSION: VApal ophthalmic solution (500 IU/mL) is safe and effective for the treatment of patients with dry eye.

Facile synthesis and properties of dithieno[3,2-b:2',3'-d]arsoles.

Dithieno[3,2-b:2',3'-d]arsoles have been synthesized via a safe and easy synthetic procedure, in which volatile arsenic intermediates are excluded. The obtained dithienoarsole derivatives were stable in the ambient atmosphere, unlike their phosphorus analogues, dithienophospholes. The Suzuki-Miyaura coupling reaction is applicable for structural modification and expansion of the π-conjugated system, and carried out the polymerization of the compounds. Dithienoarsoles showed an intense emission not only in solution but also in the solid state, and their molecular packing was analyzed by X-ray crystallography. The main-chain type dithienoarsole polymer formed a luminescent film. This work has demonstrated that dithienoarsole is a promising building block for luminescent materials.

A practical method for the generation of organoarsenic nucleophiles towards the construction of a versatile arsenic library.

Nucleophilic arsenic reagents were prepared in situ from a nonvolatile cyclooligoarsine. As-As bond cleavage of the cyclooligoarsine readily proceeded with anion sources. Various kinds of organoarsenic compounds were easily constructed in high yields by selecting anion sources and electrophiles. In comparison with conventional methods of As-C bond formation, a wide variety of organoarsenic compounds were safely and easily synthesized by using this method.

Practical Synthesis and Properties of 2,5-Diarylarsoles.

2,5-Diarylarsoles were easily synthesized from nonvolatile arsenic precursors. Diiodoarsine was generated in situ and reacted with titanacyclopentadienes to give 2,5-diarylarsoles. The structures and optical properties were studied in comparison with those of 2,5-diarylphosphole. It was found that the arsoles were much more stable in the air than the phosphole. Single crystal X-ray diffraction revealed the arsenic atoms adopted a trigonal pyramidal structure, reflecting on the s-character of the lone pair. The obtained 2,5-diarylarsoles and 2,5-diarylphosphole showed intense emission in solutions and solid state. In addition, the optical properties were controlled by transition-metal coordination.

Polymorph control of luminescence properties in molecular crystals of a platinum and organoarsenic complex and formation of stable one-dimensional nanochannel.

The mononuclear diiodoplatinum(II) complex (trans-PtI2(cis-DHDAMe)2), where cis-DHDAMe = cis-1,4-dihydro-1,4-dimethyl-2,3,5,6-tetrakis(methoxycarbonyl)-1,4-diarsinine, forms three different crystalline polymorphs that can be either concomitantly or separately obtained on varying the recrystallization conditions. Cubic red crystals (α-phase) and red-orange needles (ß-phase) exhibit solid-state red emissions at room temperature. Cubic red crystals of the γ-phase show no solid-state emission at room temperature. All crystalline structures were confirmed by X-ray crystallography. Room-temperature strongly luminescent crystals (α-phase) (λem = 657 nm, Φ = 0.52) have a triclinic P1 (No. 2) structure and no voids in the crystal structure. Red-orange needle-shaped crystals of the ß-phase exhibit moderate red luminescence (λem = 695 nm, Φ = 0.09) at room temperature and have a trigonal, R3 (No. 148), structure. In the needlelike crystals of the ß-phase, stable hexagonal arrays of nanoporous channels, 5.0 Å in diameter, are formed. Room-temperature nonluminescent crystals (γ-phase) have an orthorhombic, Pbca (No. 61), structure with a void volume that is 4.9% of the total crystal volume. After heating the α-phase crystals at 150 °C for 2 min, a powder XRD pattern different from the original crystal is obtained, and its solid-state emission at room temperature decreased. After heating the ß-phase crystals at 150 °C for 2 min, the emission wavelength and the quantum yield of the solid-state emission at room temperature and the powder XRD pattern are the same as those of the α-phase after heating at 150 °C. A crystal-to-crystal transition triggered by the thermal stimulus produces a different stable polymorph of the mononuclear diiodoplatinum(II) complex. The one-dimensional nanoporous crystals encapsulated iodine without distorting the crystal packing.

Organic vapor triggered repeatable on-off crystalline-state luminescence switching.

A nonporous crystalline solid consisting of an organoarsenic platinum(II) complex, i.e., a mononuclear diiodoplatinum(II) complex trans-PtI(2)(cis-DHDAtBu)(2) (1) with cis-1,4-dihydro-1,4-dimethyl-2,3,5,6-tetrakis(tert-butoxycarbonyl)-1,4-diarsinine (cis-DHDAtBu), shows on-off solid-state luminescence switching through reversible solvent vapor uptake and escape. The on-off switching of solid-state luminescence was achieved without changing the structure or electronic state of the organoarsenic platinum(II) complex.

The effects of Helicobacter pylori eradication on body mass index and dyspeptic symptoms.

BACKGROUND AND AIM: The relationship between Helicobacter pylori infection and body mass index (BMI) and dyspeptic symptoms is controversial. We investigated the changes in BMI and dyspeptic symptoms after H. pylori eradication among stages of atrophic gastritis classified according to the serum pepsinogen (PG) I/II ratio. METHODS: One hundred and sixty-three H. pylori-positive patients underwent eradication therapy for H. pylori. Serum PG I and II concentrations were measured before treatment, and the PG I/II ratio was classified into three groups: PG I/II ratio <2.0, PG I/II ratio >or=2.0 and <4.0, and PG I/II ratio >or=4.0 were considered to be low, middle, and high, respectively. Their BMI and abdominal symptoms were checked before, 1 and 5 years after treatment, and these changes were investigated among the three groups. RESULTS: The mean BMI changes 1 year after treatment in the low PG I/II ratio group were significantly higher than those in other groups. Most abdominal symptoms in the high PG I/II ratio group were most severe before eradication but improved significantly after eradication. CONCLUSIONS: The effects of H. pylori eradication on BMI and dyspeptic symptoms may be different according to the serum PG I/II ratio before eradication.

[A case of monoclonal gammopathy with corneal stroma deposits].

BACKGROUND: Monoclonal gammopathy is a group of disorders characterized by proliferation of a single clone of plasma cells that produce monoclonal protein. Sometimes benign monoclonal gammopathy that is a symptomatic can turn into a malignancy like multiple myeloma. We present a case of monoclonal gammopathy with corneal deposits which was treated with deep lamellar keratoplasty (DLKP). The corneal button obtained at the time of DLKP was studied with both light and electron microscopy. CASE: Fine corneal deposits developed bilaterally in a 69-year-old man. There was no family history of ocular diease. RESULTS: We diagnosed monoclonal gammopathy in a blood test that showed elevation of serum immunoglobulin G and the Kappa chain. The corneal button obtained during DLKP was analyzed histologically and ultrastructurally. Electron microscopy showed electron-dense deposits composed of fine parallel filaments in the corneal epithelium and stroma. CONCLUSIONS: Corneal deposits of unknown origin might turn into monoclonal gammopathy that could be a life-threatening disease. It is important for ophthalmologists to check the whole body of a patient when finding corneal deposits.

Immunohistochemical characterization of epithelial cells implanted in the flap-stroma interface of the cornea.

PURPOSE: To investigate the expression of extracellular matrix collagens and their relationship to corneal opacities after implantation of epithelial cells in the flap-stroma corneal interface. METHODS: A corneal flap was made on rabbit eyes, and epithelial cells, mechanically scraped from tissue surrounding the flap, were implanted beneath the flap. The corneas were harvested 1, 3, 7, and 30 days following surgery. Histological and immunohistochemical examinations were performed. The expression and localization of types I, III, and IV collagens and gelatinase A were determined. RESULTS: Slit-lamp examination showed corneal opacity in the area where the epithelial cells were implanted. Histological study revealed clusters of epithelial cells between the flap and stromal interface. One week and 1 month after the implantation, intense immunoreactivity for collagen type IV was detected at the perimeters of the intrastromal epithelial islands, but not in the interface outside the implanted epithelial cells. Weak positive staining for gelatinase A was detected in the implanted epithelial cells and surrounding keratocytes. CONCLUSIONS: The heavy deposition of collagen type IV surrounding the implanted epithelial cells indicated that it might be an essential component of the interface haze observed in patients following laser in situ Keratomileusis. Gelatinase A may also play a role in the regulation of stromal remodeling after epithelial ingrowth.

Analysis of COL8A2 gene mutation in Japanese patients with Fuchs' endothelial dystrophy and posterior polymorphous dystrophy.

PURPOSE: To determine whether Japanese patients with Fuchs' endothelial corneal dystrophy (FECD) and posterior polymorphous dystrophy (PPMD) carry mutations in the COL8A2 gene, and to investigate the possible pathogenicity of the COL8A2 gene in these corneal dystrophies. METHODS: DNA analysis of the COL8A2 gene was performed in 15 unrelated Japanese patients with FECD, and 5 patients with PPMD using polymerase chain reaction and direct sequencing. Mutation screenings were also performed in 36 unrelated normal volunteers as controls, as well as slit-lamp and specular microscopy. RESULTS: Two types of heterozygous missense mutations of the COL8A2 gene (R155Q and T502M) in 5 of 15 FECD probands (R155Q, 3/30 chromosomes, 10.0%; T502M, 3/30 chromosomes, 10.0%) were found. No mutation was detected in the coding region of the COL8A2 gene in the remaining 10 patients with FECD nor in any of the 5 patients with PPMD. These two mutations were also found in normal Japanese volunteers (R155Q, 5/72 chromosomes, 6.9%; T502M, 11/70 chromosomes, 15.7%). The chromosomal frequency of the two mutations was not significant between the patients and normal controls. CONCLUSIONS: The R155Q and T502M mutations of COL8A2 may not be the causative defect in the Japanese FECD and PPMD patients examined in this study.

Anti-angiogenic role of angiostatin during corneal wound healing.

The purpose of this study is to determine whether angiostatin is involved in maintaining corneal avascularity after wounding. We generated polyclonal rabbit anti-mouse angiostatin antibodies directed against each of the five kringle domains, (K1-5) and anti-mouse plasmin B chain antibodies. Mouse corneas were immunostained with anti-K1 angiostatin antibody after excimer laser keratectomy. Corneal epithelial cell lysate was harvested and angiostatin was isolated using lysine sepharose. Purified plasminogen was incubated with lysate of mouse corneal epithelial cells from wild type mice in the presence or absence of MMP inhibitors. Angiostatin activity was determined using calf pulmonary artery endothelial (CPAE) cell proliferation assay with and without angiostatin immunoprecipitation; and corneal neovascularization was assayed by intrastromal injection of anti-plasminogen, anti-K1-3 or anti-B chain antibodies after corneal wounding. Using the anti-mouse angiostatin antibodies that we generated, we confirmed that angiostatin-like molecules were expressed in the corneal epithelium and in cultured corneal epithelial cells. Western blotting after incubation of scraped corneal epithelial cell lysate with purified plasminogen showed reduction of the plasminogen bands at 6, 12, and 24 hr, respectively. Complete cleavage of plasminogen occurred by 48 hr. Functional assays in which corneal epithelial cell extracts were incubated with CPAE cells resulted in inhibition of vascular endothelial cell proliferation. Depletion experiments using anti-angiostatin (K1) antibodies resulted in a 25 +/- 1.2% increase in vascular endothelial cell proliferation as compared to 12 +/- 1.8% using the protein A control (p < 0.05). Corneal neovascularization was observed after excimer laser keratectomy when anti-angiostatin antibodies were injected into the cornea (65 +/- 13%) which was significantly higher than when plasmin B chain antibodies were injected (10 +/- 2.6%; p < 0.05). Plasminogen and angiostatin are produced in the cornea. They may play a role in preventing vascularization and may contribute to the maintenance of corneal avascularity after excimer laser keratectomy.

Expression of type XVIII collagen during healing of corneal incisions and keratectomy wounds.

PURPOSE: To determine the distribution of type XVIII collagen in mouse ocular tissues and to investigate the expression of type XVIII collagen during healing of corneal incisions and keratectomy wounds. METHODS: Immunohistochemical analysis of type XVIII collagen was performed in mouse ocular tissue, with polyclonal antibodies to the hinge domain. For wound-healing experiments, excimer laser keratectomy and single linear incisions were performed on mouse corneas. The corneas were harvested at various time points after wounding and processed for immunohistochemistry, in situ hybridization, competitive reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analysis. RESULTS: In the unwounded mouse cornea, type XVIII collagen was expressed by the corneal epithelial cells. Type XVIII collagen was immunolocalized to the mouse corneal epithelium, epithelial basement membrane, Descemet's membrane, ciliary epithelium, lens capsule, retinal inner limiting membrane, and Bruch's membrane. In the early stages of wound healing after excimer laser keratectomy (days 3 and 7), type XVIII collagen staining of the epithelial basement membrane was absent, whereas its localization to Descemet's membrane was unchanged. After linear corneal incisions, however, type XVIII collagen was clearly seen in the stroma and in the epithelial basement membrane. Type XVIII collagen immunolocalization to the subepithelial stromal wound region peaked at 1 week after wounding, and its mRNA showed a corresponding temporal increase in expression within the same region after linear corneal incisions. CONCLUSIONS: The results suggest that type XVIII collagen is broadly expressed in ocular tissues and that it may have a role in wound healing, especially after incisional corneal wounds.

Corneal neovascularization after excimer keratectomy wounds in matrilysin-deficient mice.

PURPOSE: Matrilysin, matrix metalloproteinase (MMP)-7, is upregulated in the corneal epithelium during wound healing after excimer keratectomy wounds. The purpose of this study was to determine the role of matrilysin in maintaining corneal avascularity during wound healing. METHODS: Matrilysin-deficient mice (n = 17) and their age-matched wild-type littermates (n = 18) were treated with 193 nm argon-fluoride excimer keratectomy (experiment I). The percentage of corneal surface occupied by neovascularization was measured with a computer image-analysis program adjusted for parallax. In another experiment (experiment II), epithelial closure was monitored with slit lamp biomicroscopy and fluorescein staining, and corneal neovascularization was confirmed by india ink perfusion, electron microscopy, and immunolocalization of CD31 and type IV collagen. Corneal micropocket assays were performed to compare the area of corneal neovascularization in matrilysin-deficient mice and wild-type littermates (experiment III). To determine whether the differences in corneal neovascularization were related to differences in angiogenic factors, the levels of basic fibroblast growth factor (bFGF) were compared with those of vascular endothelial growth factor (VEGF) in matrilysin-deficient and wild-type mouse corneas (experiment IV). RESULTS: The percentages of the corneal surface occupied by neovascularization after excimer laser keratectomy in the matrilysin-deficient mice measured 21.3% +/- 5.2% and 18.7% +/- 5.8% at days 3 and 7, respectively, compared with 5.3% +/- 2.4% and 5.5% +/- 3.4% in the wild-type littermates at days 3 (P < 0.01) and 7, respectively (P < 0.05; experiment I). No significant differences in the rates of epithelial closure of corneal wounds were observed between matrilysin-deficient and wild-type mice after wounding. Corneal neovascularization in the matrilysin-deficient mice was confirmed by india ink present in the corneal stromal blood vessels (extending from the limbus to the wound), immunohistochemical staining, and electron microscopy. Gram, Giemsa, calcofluor white, and acridine orange stains and electron microscopy showed no evidence of corneal infection (experiment II). The area of corneal neovascularization in matrilysin-deficient mice was not significantly different from that of wild-type littermates after implantation of bFGF pellets (0.91 +/- 0.55 mm(2) and 0.77 +/- 0.34 mm(2), respectively; experiment III). The levels of bFGF and VEGF (VEGF, VEGF-B, and VEGF-C) in corneal epithelial cells were not elevated in matrilysin-deficient mice compared with the wild-type mice (experiment IV). CONCLUSIONS: Matrilysin may play an important role in maintaining corneal avascularity during wound healing. The differences in corneal neovascularization between matrilysin-deficient mice and wild-type littermates seem unrelated to the bFGF and VEGF levels in the corneal epithelium.

Confocal and electron microscopic studies of laser subepithelial keratomileusis (LASEK) in the white leghorn chick eye.

OBJECTIVES: To evaluate the effect of 20% alcohol on the white leghorn chick cornea and to determine the confocal and electron microscopic findings of laser subepithelial keratomileusis surgery in the white leghorn chick corneal model. METHOD: Laser subepithelial keratomileusis surgery was performed on chick corneas and the morphologic changes were examined by transmission electron microscopy. Chick corneas were exposed to 20% alcohol for 30 and 45 seconds or 1 and 2 minutes (5 chicks per group) to evaluate the effect on the corneal epithelium. Photorefractive keratectomy using either mechanical or 20% alcohol-assisted debridement (5 chicks per group) was also performed. Keratocyte and epithelial cell deaths were analyzed 4 hours after surgery using terminal deoxynucleotidyl transfer-mediated biotin-dexoyuridine 5-triphosphate nick-end labeling (TUNEL) staining and transmission electron microscopy. RESULTS: Exposure of the corneal epithelium to 20% alcohol for 30 seconds or longer allowed reproducible separation of epithelial flaps in white leghorn chick eyes. Transmission electron microscopy immediately after alcohol treatment showed that exposure to 20% alcohol for 30 seconds or less had minimal adverse effects on the corneal epithelium. The TUNEL staining of corneas obtained 4 hours after surgery revealed TUNEL-positive cells in the central superficial stroma and more abundantly in the peripheral superficial stroma around the epithelial flap margin and in the epithelial flap itself, particularly in the basal epithelial layer. Transmission electron microscopy showed similar evidence of apoptosis in the epithelium and anterior stroma. CONCLUSIONS: The white leghorn chick eye seems to be a reasonable model for laser subepithelial keratomileusis surgery. Treatment with 20% alcohol for 30 seconds results in reproducible epithelial flap creation in the chick cornea and in relatively low levels of stromal and epithelial cell death after surgery.