Simultaneous Determination of Nine Phthalates in Vegetable Oil by Atmospheric Pressure Gas Chromatography with Tandem Mass Spectrometry (APGC-MS/MS).

Although the use of phthalates has been restricted worldwide, they remain an issue due to health concerns. Diet is one of the most important exposure pathways for humans and due to their solubility in oil, phthalates are commonly found in edible oil and food high in fat. Gas chromatography-mass spectrometry (GC-MS) using electron ionization (EI) has been commonly used for the analysis of the phthalates in foodstuffs, including edible oil. However, this method suffers from issues with sensitivity and selectivity, as most phthalates are fragmented to generate a common phthalic anhydride fragment ion at m/z 149. The molecular ion cannot be observed due to strong fragmentation in EI. In contrast, atmospheric pressure gas chromatography (APGC) is a soft ionization technique with less fragmentation, whereby the molecular ion can be used as the precursor ion for multiple reaction monitoring (MRM). In this study, a simple and quick method for the determination of phthalates in vegetable oil using APGC-MS/MS was developed, and performance was assessed. The method was based on dilution of the oil in solvent and direct injection without the need for further cleanup. The established method was evaluated for linearity, recovery, precision, method detection limit (MDL), and method quantitation limit (MQL). The obtained MQL in vegetable oil was in the range of 0.015-0.058 mg/kg, despite limiting the injection volume to 1 µL, which is suitable for investigating dietary exposure and future proof against decreases to the regulatory limit. Finally, the developed method was successfully applied to analyze nine phthalates in eight commercially available vegetable oil.

Efficacies of preoperative prism adaptation test and monocular occlusion for detecting the maximum angle of deviation in intermittent exotropia.

BACKGROUND: The efficacies of prism adaptation test (PAT) and monocular occlusion (MO) and their optimal test durations to detect the maximum angles of deviation at near and distance in eyes with intermittent exotropia (IXT) were assessed and compared. METHODS: We retrospectively reviewed the medical records of 72 patients with IXT. All the patients had undergone the initial strabismus surgery between April 2015 and October 2018 and had been preoperatively tested by both PAT and MO performed on different days for 30 and 60 min. Near and distance deviations after 30 and 60 min of PAT and MO were compared to their baseline measurements obtained immediately after prism wear and before occlusion by alternate prism cover test. The near/distance measurements and required test duration to reveal the maximum deviation angle were also compared between PAT and MO. RESULTS: Compared with the baseline, the near deviation by PAT significantly increased after 30 (P < 0.05) and 60 (P < 0.01) minutes but not the distance deviation. However, the increase after 30 min was not significant. By MO, neither near nor distance deviation showed a significant difference from the baseline after 30 and 60 min. PAT showed a significantly larger near deviation than MO at 30 and 60 min, but a larger distance deviation by PAT was only observed at 30 min. CONCLUSIONS: In patients with basic and convergence insufficiency types of IXT, a 30-minute PAT appears to be more effective than MO in revealing the maximum angle of deviation before strabismus surgery.

Efficacy of toric intraocular lens and prevention of axis misalignment by optic capture in pediatric cataract surgery.

PURPOSE: To compare the outcomes of intraocular lens (IOL) implantation using toric (T)-IOL and nontoric (N)-IOL in pediatric cataract patients with astigmatism and to examine the effect of optic capture (OC) on the axis misalignment (AM) of the T-IOLs. SETTING: Department of Ophthalmology, Kindai University Hospital, Osaka, Japan. DESIGN: Interventional, comparative case study. METHODS: Consecutive pediatric patients implanted with T-IOLs or N-IOLs were retrospectively reviewed. In the T-IOL group, the preoperative and postoperative corrected distance visual acuity (CDVA) and AM were compared in patients with and without OC. RESULTS: The T-IOL group included 14 eyes of 11 patients, whereas the N-IOL group included 22 eyes of 15 patients. One year postoperatively, the mean ocular cylinder (1.38 ± 0.80 diopters [D]) was significantly smaller than the mean corneal cylinder (3.33 ± 1.24 D) in the T-IOL group (P = .00012, Wilcoxon signed-rank test). The mean preoperative and 1-year postoperative CDVA (logMAR) were 0.57 and 0.003 (T-IOL) and 0.71 and 0.09 (N-IOL), respectively. The AM at 1 week and 1 year postoperatively was 2.6 ± 3.7 degrees and 4.4 ± 3.1 degrees for the OC group, and 13.3 ± 8.8 degrees and 18.5 ± 14.8 degrees, for the non-OC group, respectively. The AM was significantly smaller in the OC group than that in the non-OC group (P = .009, Mann-Whitney U test) at postoperative 1 week. CONCLUSIONS: T-IOL implantation is effective in correcting astigmatism in pediatric cataract patients with astigmatism, and the OC technique is likely to achieve lower AM of the T-IOL.

Role of fusional convergence amplitude in postoperative phoria maintenance in children with intermittent exotropia.

PURPOSE: To examine the role of fusional convergence amplitude in postoperative phoria maintenance in childhood intermittent exotropia [X(T)]. METHODS: The medical records of 29 children aged 15 years or younger (mean age, 10.8 ± 2.4 years) and treated with monocular recession-resection for X(T) were reviewed retrospectively. The patients' fusional convergence amplitude (break point/total amplitudes), physiologic diplopia, and phoria maintenance (presence/absence of phoria maintenance and ability to maintain phoria) were assessed. The presence of phoria maintenance was confirmed by a cover test, and the ability to maintain phoria was quantified using the Bagolini red filter bar. Correlations of the amplitude size with the presence and ability of phoria maintenance were investigated. RESULTS: A significant correlation was seen between fusional amplitude (break point/total) and ability to maintain phoria at near and at far (break point: P < .05 at near/P < .01 at far; total: P < .05 at near/far). Neither the break point amplitude nor the total amplitude significantly differed between the patients with phoria maintenance and those without it (break point: P = .71 at near, P = .29 at far; total: P = .98 at near, P = .85 at far). Phoria maintenance correlated with the suppression of physiologic diplopia during phoria (P < .01). The deviation angle did not significantly correlate with fusional amplitude either at near (P = .58) or at far (P = .27). CONCLUSIONS: In childhood X(T), fusional amplitude plays a role in enforcing the patient's ability to maintain phoria. However, sufficient fusional amplitude does not guarantee fully functioning fusion if suppression is present during phoria.

Influence of Test Distance on Stereoacuity in Intermittent Exotropia.

AIMS: To investigate influence of test distance on stereoacuity in intermittent exotropia (X[T]) using the same test conditions for both near and far distances. METHODS: Subjects were 38 consecutive patients with X(T). All the patients were between ages 6 and 15 years and had decimal visual acuity of 1.0 or better. Another inclusion criterion was presence of phoric condition at near and far distances. Stereoacuity was measured at a near distance of 40 cm and at a far distance of 5 m. The following test conditions were used for both test distances: separation of the two eyes using polarized glasses, and a target with a random dot pattern. All the stereograms had the same subtended angle of 2.5º, and binocular disparity of 480, 240, 120, and 60 arcsec. We used two stereogram types with crossed and uncrossed disparities. RESULTS: Far stereoacuity of 38 subjects measured with the crossed disparity was significantly worse than near stereoacuity (P<0.05, Wilcoxon signed-ranks test), although 30 (78.9%) of the 38 subjects showed no differences in stereopsis between the near and far distances. Far stereoacuity of 38 cases measured with the uncrossed disparity was significantly worse than at near (P<0.05, Wilcoxon signed-ranks test), although 20 (52.6%) of the 38 subjects showed no differences between stereoacuity at near and far. In comparison of stereoacuity with crossed disparity and uncrossed disparity, stereoacuity with crossed disparity was significantly better than that with uncrossed disparity both at near and far (P<0.05, Wilcoxon signed-ranks test). CONCLUSIONS: Stereoacuity in X(T) was different according to test distance when measured controlling subtended angle of stereogram at both distances. Far stereoacuity was significantly worse than near stereoacuity when measured using test targets with both crossed and uncrossed disparities. Additionally, stereoacuity measured with crossed disparity was better than that with uncrossed disparity at both distances.

Optical coherence tomographic findings at the fixation point in a case of bilateral congenital macular coloboma.

BACKGROUND: Congenital macular coloboma is a rare ocular disease that consists of atrophic lesions in the macula with well-circumscribed borders. We report the findings of spectral domain optical coherence tomography (SD-OCT) at the fixation point in a case of bilateral macular coloboma. CASE REPORT: The subject is a 4-year-old boy. He visited our hospital at age 1 year and 4 months for the evaluation of strabismus. The fundus examination of both eyes showed round-shaped sharply-demarcated atrophic lesions involving the macula with large choroidal vessels and bared sclera at the base. Immunologic tests including toxoplasmosis, rubella, varicella, herpes virus, and human T-cell leukemia virus were all negative. At age 4 years and 1 month, cycloplegic refraction showed insignificant refractive errors and his best corrected visual acuity was 0.6 bilaterally. The SD-OCT showed a crater-like depression accompanying atrophic neurosensory retina, and the absence of retinal pigment epithelium and choroid. Examination of the fixation behavior by visuscope showed steady fixation with an area 0.5° nasal to the nasal edge of the atrophic lesion bilaterally. The SD-OCT findings at fixation area showed remaining normal retinal structures involving inner segment-outer segment (IS/OS) junction line. CONCLUSION: The findings of SD-OCT have been shown to be useful in the diagnosis of macular coloboma. In the fixation point, the structure of retina and choroid were well preserved.

Longitudinal clinical course of three Japanese patients with Leber congenital amaurosis/early-onset retinal dystrophy with RDH12 mutation.

PURPOSE: To report the longitudinal clinical course of three Japanese patients from two families with Leber congenital amaurosis/early-onset retinal dystrophy (LCA/EORD), and the results of next-generation DNA sequences on them. PATIENTS AND METHODS: The patients were three Japanese children: a 4-year-old girl, a 6-year-old boy, and a 3-year-old girl. Patients 1 and 2 were siblings, and patient 3 was from an unrelated family. Standard ophthalmic examinations including perimetry, electroretinography, optical coherence tomography, and ultrasonography were performed on each patient. The patients were observed for 28, 16, and 10 years. Whole exomes of the patients and their non-symptomatic parents were analyzed using a next-generation sequence technique. RESULTS: The decimal visual acuity varied between 0.07 and 0.6 at the initial visit and decreased to counting finger to hand motion in their teens. Funduscopy showed diffuse retinal and macular degeneration. During the follow-up period, a posterior staphyloma developed and the macular area became atrophic. Patient 1 developed cataracts in her early twenties. Genetic analysis revealed a homozygous A126V substitution in the RDH12 gene in all patients. CONCLUSIONS: The three patients with LCA/EORD had a progressive decrease of their vision with the formation of a posterior staphyloma. This is the first report of Japanese patients with LCA/EORD with a RDH12 mutation.

Effect of suppression during tropia and phoria on phoria maintenance in intermittent exotropia.

BACKGROUND: Treatment for intermittent exotropia X(T) aims to keep the eye in a phoric position and to maintain the phoria. However, maintenance of phoria is difficult even after treatment, and the cause is unclear. The aim of this study was to investigate the presence of suppression during tropia and/or phoria in X(T), and to determine how the suppression affected patient's ability to maintain phoria. METHODS: Medical records of 89 children with X(T) (mean age, 9.8 ± 2.7 years) were reviewed retrospectively. According to their previous treatment for X(T), the patients were divided into four groups and compared: untreated and under observation only (28 patients), surgical treatment (32 patients), orthoptic training (eight patients) and a combined treatment of surgery and orthoptic training (21 patients). Suppression during phoria was evaluated by a physiologic diplopia test, and suppression during tropia was evaluated by a convergence test or a cover test when fusion broke. Phoria maintenance was achieved if a phoric condition was maintained even when the fusion broke at both near and far. Furthermore, the Bagolini's red filter bar was used to quantitatively assess patient's ability to maintain phoria at near and far distances. RESULTS: No subject only suppressed during phoria. Patients who suppressed under both conditions could not maintain phoria. Suppression under both conditions significantly correlated with phoria maintenance and the ability to maintain phoria (P < 0.01, Fisher's exact probability test). All the patients with a strong ability to maintain phoria did not suppress under either condition. As compared to the surgical treatment group, the combined treatment group had a higher percentage of patients who did not suppress under either condition and could maintain the phoria. Suppression under both conditions also significantly correlated the treatment methods (P < 0.01, Chi-square for the independence test). CONCLUSIONS: Suppression under both tropic and phoric conditions significantly relates to the outcome of patients' phoria maintenance and their ability to maintain a phoric position. Suppression under both conditions is an important indication of whether X(T) shifts to constant exotropia.

Slug is upregulated during wound healing and regulates cellular phenotypes in corneal epithelial cells.

PURPOSE: The involvement of the epithelial mesenchymal transition (EMT) in the process of corneal wound healing remains largely unclear. The purpose of the present study was to gain insight into Slug expression and corneal wound healing. METHODS: Slug expression during wound healing in the murine cornea was evaluated using fluorescence staining in vivo. Slug or Snail was stably introduced into human corneal epithelial cells (HCECs). These stable transfectants were evaluated for the induction of the EMT, cellular growth, migration activity, and expression changes in differentiation-related molecules. RESULTS: Slug, but not Snail, was clearly expressed in the nuclei of corneal epithelial cells in basal lesion of the corneal epithelium during wound healing in vivo. The overexpression of Slug or Snail induced an EMT-like cellular morphology and cadherin switching in HCECs, indicating that these transcription factors were able to mediate the typical EMT in HCECs. The overexpression of Slug or Snail suppressed cellular proliferation but enhanced the migration activity. Furthermore, ABCG2, TP63, and keratin 19, which are known as stemness-related molecules, were downregulated in these transfectants. CONCLUSIONS: It was found that Slug is upregulated during corneal wound healing in vivo. The overexpression of Slug mediated a change in the cellular phenotype affecting proliferation, migration, and expression levels of differentiation-related molecules. This is the first evidence that Slug is regulated during the process of corneal wound healing in the corneal epithelium in vivo, providing a novel insight into the EMT and Slug expression in corneal wound healing.

Successive iterative restoration applied to streak artifact reduction in X-ray CT image of dento-alveolar region.

PURPOSE: X-ray computed tomography (CT) images in the dento-alveolar region are sometimes rendered unusable for diagnostic purposes due to the appearance of streak artifacts. The purpose of the study is to reduce streak artifacts appeared on dental and maxillofacial X-ray CT images by the application of modified iterative restoration method. METHODS: We took advantage of the aspect that adjacent CT images often depict very similar anatomical structures within the resulting collection of thin-slice images. CT images having streak artifacts were processed using the projection data of adjacent CT images. A modified iterative correction, the maximum likelihood-expectation maximization (ML-EM) reconstruction algorithm, was employed to reduce the streak artifact caused by metallic materials in the oral cavity. It approximates between the processed image and the original projection data. First, the projection data of an intact image were obtained, and then, the next image that had streak artifacts was processed. The projection data of the processed image were obtained, and the ML-EM method was applied to the next image again. Then, the successive iterative restoration was carried out. RESULTS: Twelve adjacent images were processed. Each iterative restoration was carried out fifty times. Streak artifacts were observed on processed images at the initial stage, but some of them either suppressed or disappeared as the iteration progressed. CONCLUSIONS: The modified ML-EM method was effective to reduce streak artifacts in X-ray CT images in dento-alveolar region.

Connective tissue growth factor cooperates with fibronectin in enhancing attachment and migration of corneal epithelial cells.

Corneal wound healing is a complex process involving the integrated actions of various growth factors, cytokines and extracellular matrix produced by corneal cells and inflammatory cells. Connective tissue growth factor (CTGF) has been linked to wound healing, and fibronectin (FN) is a major component of the extracellular matrix. However, the functions of CTGF and FN in corneal epithelial cells are not well understood. We therefore investigated the coordinated function of CTGF and FN in the attachment and migration of corneal epithelial cells. Treatment of human corneal epithelial cells (HCECs) with transforming growth factor (TGF) beta1 up-regulated the expression of CTGF, but did not noticeably affect FN expression, as judged by immunoblot analysis of cell lysates. In contrast, the amount of FN accumulated in the cultured media was increased in a time-dependent manner, but CTGF was undetectable in the cultured media. The expression level of FN was decreased by the knockdown of CTGF expression with a specific short hairpin RNA, indicating that CTGF acts as an upstream mediator of FN expression. CTGF augmented the FN-mediated increase in the attachment of HCEC by about twofold, although CTGF alone did not influence the attachment. Moreover, the migration assay with rabbit corneal blocks revealed that CTGF (390 nM) alone or in combination of FN (10 microg/mL) promoted corneal epithelial migration; the mean migration distances of control, CTGF, and CTGF + FN were 272, 325, and 626, microm, respectively. In conclusion, CTGF cooperates with FN in enhancing the attachment and migration of corneal epithelial cells.

Analysis of Gibbs monolayer adsorbed at the toluene/water interface by UV-visible partial internal reflection spectrometry.

Gibbs monolayers of lipophilic tetraphenylporphyrinatomanganese(III) and hydrophilic diacid of meso-tetrakis(4-sulfonatopheny)porphyrin adsorbed at the liquid-liquid interface have been analyzed by UV-visible external reflection (ER) and partial internal reflection (PIR) spectra measured at different angles of incidence. The angle-dependent ER and PIR spectra over the Brewster angles (thetaERB and thetaIRB) have readily been measured at the toluene/water interface. As preliminarily expected in our previous study, the present study has first proved that the reflection-absorbance of UV-visible PIR spectra quantitatively agrees with the theoretical calculations for the Gibbs monolayer over thetaIRB. In addition, it has also been proved that the absorbance of the PIR spectra is greatly enhanced in comparison to that of the ATR spectra. The enhancement is caused by an optical effect in the monolayer sandwiched between two phases of toluene and water that have different but refractive indices close to each other. This optical enhancement requires an optically perfect contact between the phases, which is difficult to prepare for a solid-solid contact. At the liquid/liquid interface, however, an ideal optical contact is easily realized, which makes the enhancement as much as the theoretical expectation. The PIR spectrometry will be recognized to be a new high-sensitive analytical tool to study Gibbs monolayer at the liquid/liquid interface.

The cytokine regulation of SPARC production by rabbit corneal epithelial cells and fibroblasts in vitro.

OBJECTIVE: SPARC (osteonectin/BM40) is detected in the corneal stroma during the wound-healing process. To understand the metabolism of SPARC in the cornea, we investigated the effects of cytokines and growth factors on SPARC synthesis by rabbit corneal epithelial cells and fibroblasts. METHODS: Rabbit corneal epithelial cells or fibroblasts were cultured for 3 days with serum-containing minimal essential medium (MEM), then subcultured for 3 days on serum-free MEM with epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factor-beta (TGF-beta), or interleukin-1beta (IL-1beta). SPARC concentration in the medium was measured by the ELISA method using anti-SPARC monoclonal antibody. RESULTS: The concentration of SPARC in the conditioned medium of the epithelial cells depended on either cell numbers or cultivation periods. When EGF was added to the medium, the amount of SPARC in the medium decreased. The addition of IL-1beta, PDGF, or TGF-beta did not affect SPARC synthesis by the epithelial cells. The production of SPARC by rabbit corneal fibroblasts was low compared with that by epithelial cells. However, the synthesis of SPARC by corneal fibroblasts was significantly enhanced by the addition of TGF-beta. The addition of IL-1beta, PDGF, or EGF slightly increased SPARC synthesis by corneal fibroblasts. CONCLUSIONS: Cytokines and growth factors modulate SPARC synthesis by rabbit corneal epithelial cells and fibroblasts. These results suggest that cytokines and growth factors modulate cell-matrix interaction in corneal wound healing, possibly by regulating SPARC synthesis.