Biomedical subjects

Mitsuru Sawa

Publications and source records attributed to Mitsuru Sawa.

At least 19 recordsLinked to original sources

PubMed2006

Effect of surface coating an acrylic intraocular lens with poly(2-methacryloyloxyethyl phosphorylcholine) polymer on lens epithelial cell line behavior.

PURPOSE: To evaluate the effect of surface coating of an acrylic intraocular lens (IOL) with poly(2-methacryloyloxyethyl phosphorylcholine) (MPC) on the behavior of the lens epithelial cell (LEC) line, alpha-TN4. SETTING: Department of Ophthalmology, Nihon University School of Medicine, Tokyo, Japan. METHODS: A hydrophobic soft acrylic IOL (AF-1, Hoya) was coated with MPC polymer. A noncoated IOL served as control. An IOL from each group was placed on the membrane of collagen I or IV of the cell culture dish. The alpha-TN4 cells were seeded in the insert. Cell behaviors (ie, cell proliferation and spreading) on IOLs and membranes were observed. Cell migration beneath the IOL optic portion was assayed using a computer software program (POCOman system) for posterior capsule opacification (PCO). Type I or IV collagen is the major matrix component of PCO or native lens capsule. RESULTS: Cell proliferation was more marked on the noncoated IOL than on the coated IOL. Type IV collagen accelerated proliferation more than type I collagen. Cell migration to the area beneath the IOL optic was more prominent in the group with the type I collagen membrane and noncoated IOL than in other groups. CONCLUSION: Coating an acrylic IOL surface with MPC polymer suppressed adhesion and proliferation of LECs, suggesting it improves IOL biocompatibility.

Acrylic Resins↗

PubMed2006

Efficacy of levocabastine hydrochloride ophthalmic suspension in the conjunctival allergen challenge test in Japanese subjects with seasonal allergic conjunctivitis.

BACKGROUND: This study was conducted to investigate the efficacy and safety of 0.025% levocabastine hydrochloride in Japanese subjects with seasonal allergic conjunctivitis and its duration of action using the conjunctival allergen challenge (CAC) test. METHODS: Twenty-four asymptomatic subjects were randomized to instill 0.025% levocabastine ophthalmic suspension in one eye and vehicle in the other eye 10 minutes before the CAC test. Signs and symptoms of allergic conjunctivitis were scored 10, 15, and 25 minutes after the CAC test. The duration of drug effects was also evaluated by allergen rechallenge 4 hours after levocabastine administration. The itching score for each eye as the primary efficacy endpoint was assessed 15 minutes after the CAC test using a 5-point scale. RESULTS: The mean itching score in the levocabastine-treated group was 0.08 +/- 0.06, which was significantly lower than the mean score of 1.98 +/- 0.16 in the vehicle group (P < 0.0001). The redness and chemosis of the conjunctiva were also improved significantly compared with the vehicle group. Levocabastine showed prolonged efficacy in inhibiting itching (0.42 +/- 0.12 vs 0.94 +/- 0.17, P < 0.0002) and redness (1.04 +/- 0.18 vs 1.42 +/- 0.22, P < 0.01) of the conjunctiva upon the rechallenge test. No significant topical or systemic adverse safety findings were observed in the levocabastine group. CONCLUSIONS: The results indicate that 0.025% levocabastine ophthalmic suspension is effective and safe in the treatment of allergic conjunctivitis with a duration of action of at least 4 h.

Adult↗

PubMed2006

[Tear eosinophil cationic protein measurement in patients with seasonal allergic conjunctivitis].

PURPOSE: In the patients with seasonal allergy conjunctivitis (SAC), we investigated the concentration of tear eosinophil cationic protein (ECP) levels, the diagnosis of SAC and the usefulness of studying the disease states. SUBJECTS AND METHODS: The subjects were patients suffering from conjunctivitis seasonal allergy caused by cedar pollen and a healthy control group, and the study period was three years from 2001 to 2003. We divided the allergy group into 27 eyes of 27 patients that we measured before the pollen season and 44 SAC eyes of 44 patients that we measured during the pollen season. The healthy control group was 23 eyes of 23 healthy adults. Tears were collected by the schirmer I method. The ECP value of specimens was measured by enzyme-linked immunosorbent assay (ELISA) using samples which we eluted in buffer (0.5 M NaCl + 0.5% tris buffer saline with Tween 20) from Schirmer test paper. RESULTS: The measurement range of ELISA was 3.6-180 microg/l. The mean ECP value above the lower limit of ELISA in tears was 5.06 +/- 3.39 microg/l (n = 7) (mean +/- standard deviation) for the healthy control group, 9.32 +/- 6 .68 (n = 11) for the preseason group, and 54.90 +/- 117.74 (n = 29) for the pollen season group. The difference between the healthy group and the seasonal group was highly significant (p < 0.005), but there was no significant difference between the healthy group and the preseason group. We set the cut-off point at the 95th percentile (11.1 microg/l) of the healthy group and considered any value above cut-off as positive. The ECP value was positive in 2.5% (3 eyes out of 27) in the preseason group and 40.9% (18 of 44) in the seasonal group, and there was a significant difference (p < 0.01) between the seasonal group and the healthy group. When the seasonal group was divided into patients who had started treatment before the pollen season and those who began treatment during the season, we found a small ECP difference, but it was not statistically significant. CONCLUSION: Measurement of the ECP value in seasonal pollen allergy sufferers may be useful for pathological ascertainment, but it dose not provide adequate diagnosis because of the large number of false negative cases.

Adult↗

PubMed2006

[Clinical evaluation of a measurement method for secretory IgA in tears].

PURPOSE: To evaluate the efficacy of measurement of secretory IgA (sIgA) in tears with tear sampling methods using filter paper, and to review sIgA measurement method for clinical application. SUBJECTS AND METHODS: Subjects were divided into the following 4 groups: a healthy control group, 29 eyes of 29 subjects; a contact lens group, 15 eyes of 15 subjects; a dry eye group, 13 eyes of 13 subjects; and a herpes group, 6 eyes of 6 subjects. In all subjects the sIgA value in tears was measured. In addition, in eighteen eyes of 18 healthy control individuals, the tear sIgA value was measured three times, morning, noon, and night in one day, and the variation of tear sIgA value was checked. The tears were sampled by the Schirmer 1 method. Schirmer papers were eluted in 200 microl of 0.5 M NaCl and 0.5% Tween 20 in 0.05 M phosphate-buffered solution (pH 7.2). Tear sIgA value was measured by ELISA (enzyme-linked immunosorbent assay). RESULTS: Tear sIgA value was 1249.0+/-1025.0 microg/ ml in the healthy control group; 1057.4+/-1583.3 microg /ml in the contact len groups; 197.8+/-91.3 microg/ml in the dry eye group; and 759.7+/-467.8 microg/ml in the herpes group. There was no significant difference between the healthy control group and the contact lens group, or the control group and the herpes group. There was a significantly (p<0.0001) low value in the dry eye group in comparison with the healthy control group. In the 18 healthy control individuals, there was a tendency for the tear sIgA value collected at noon to be higher. CONCLUSION: Measurment of the changes in tear sIgA values caused by inflammation of the lacrimal gland is useful as a clinical test of lacrimal gland function.

Adult↗

PubMed2006

[Evaluation of the chemokine variation in tears of contact lens wearers].

PURPOSE: To evaluate the variation of chemokines in tears of contact lens wearers. SUBJECTS AND METHODS: The subjects were divided into the three groups: a control group consisting of 26 eyes of 26 healthy volunteers without contact lenses a contact lens group (CL group) consisting of 30 eyes of 30 healthy contact lens wearers without ocular surface disorders, and a giant papillary conjunctivitis group (GPC group) consisting of 9 eyes of 9 patients with giant papillary conjunctivitis caused by contact lens wearing. Tear samples were taken by the modified Schirmer I method using a filter paper. Tear samples were eluted and analyzed for chemokines including interleukin-8 (IL-8), eotaxin-2, and pulmonary and activation-regulated CC chemokine (PARC) by the antibody array method. Concentrations of IL-8, eotaxin-2, and PARC in tears were determined quantitatively by enzyme-linked immunosorbent assay (ELISA). RESULTS: Using the antibody array method, the expression of IL-8, eotaxin-2, and PARC in the GPC group was 4-fold higher or greater than in the control group. In the measurement by ELISA, IL-8 levels in the GPC group (1154.5 +/- 1739.3 pg/ml) (mean +/- SD) were significantly higher (Kruskal-Wallis test, p < 0.01) than in the control (75.2 +/- 88.7 pg/ml) and CL (153.6 +/- 252.8 pg/ml) groups. The eotaxin-2 levels in tears did not show a statistical difference among the three groups. The PARC level in tears of the GPC group (2859.6 +/- 2299.9 pg/ml) was significantly higher than in the control(589.0 +/- 324.8 pg/ml) and CL (671.7 +/- 536.2 pg/ml) groups (Kruskal-Wallis test, p < 0.05). CONCLUSION: Wearing a contact lens per se disorders, does not cause chemokine variation in tears. However, an increase of IL-8 which induces neutrophilic invasion and an increase of PARC which induces lymphocyte invasion play an important roles in increasing the risk factor of GPC when wearing contacts.

Adolescent↗

PubMed2005

[Effects of surface modification of intraocular lenses on foreign body reaction ].

PURPOSE: In order to improve biocompatibility, we investigated the effects of surface modification by 2-methacryloyloxyethyl phosphorylcholine (MPC) on the foreign body reaction of intraocular lens (IOLs). MATERIALS AND METHODS: Materials of the IOLs were polymethylmethacrylate, hydrophobic acryl, and MPC surface-modified hydrophobic IOLs (MPC modified acryl). In an in vitro study, cultured macrophages sampled from mouse intra-abdominal exudate were cultured on a plate for each IOL material. The cell density and morphology of attached cells on the IOL materials were investigated. In an in vivo study, each IOL material was implanted in the peritoneal space of mice and foreign body reaction was investigated with a light microscope and a scanning electron microscope. RESULTS: In the in vitro study, the cells on the MPC modified acryl IOL material were remarkably fewer than those on the plates of the other two IOL materials. Regarding the implanted IOL matrevials, MPC modified acryl IOL material showed more polynuclear giant foreign body cells in the early period than the other two IOL materials. CONCLUSION: MPC surface modification can reduce the foreign body reaction of IOLs and has the potential to improve biocompatibility of IOL materials.

Animals↗

PubMed2004

[Histopathological study of lattice corneal dystrophy with L 527 R mutation of transforming growth factor-beta induced gene].

PURPOSE: A histopathological study of lattice corneal dystrophy with L 527 R mutation of exon 12 on transforming growth factor-beta induced gene(TGFBI). PATIENT AND METHODS: The patient was a 66-year-old woman with lattice corneal dystrophy. Gene analysis and histopathological study of corneal tissue obtained at the corneal transplantation were performed. Mutation analysis was done for exon 4, 11, and 12 on the TGFBI gene by direct sequencing. A histological study was performed by light microscopy with hematoxylin-eosin staining, dylon staining, or congo red staining, and by electron microscopy. RESULTS: Gene analysis revealed L 527 R mutation of exon 12 on the TGFBI gene. Light microscopic examination showed dylon-positive and Congo red-positive amyloid deposit from the superficial to deep layer of the corneal stroma. In transmission electron microscopy, the corneal epithelial layer, the basement membrane of the epithelium, Descemet's membrane, and the corneal endothelium were normal, but irregularity of Bowman's membrane and amyloid fibers in the corneal stroma were observed. CONCLUSION: Lattice corneal dystrophy due to L 527 R mutation showed histopathological findings similar to those of lattice corneal dystrophy type III.

Aged↗

PubMed2004

[Tumor necrosis factor-alpha in tears of patients with Sjögren syndrome].

PURPOSE: To determine the pathological role of tumor necrosis factor alpha (TNF-alpha) in ocular surface disorder of patients with Sjögren syndrome (SS), TNF-alpha in the tears of patients with SS were analyzed. In addition, the relationship between TNF-alpha levels in the tear samples and the severity of the corneal epithelial disorder was evaluated. MATERIAL AND METHODS: Tear samples were obtained from the 60 eyes of 30 patients with primary SS and the 60 eyes of 30 normal subjects. Tear samples were analyzed using a sandwich enzyme-linked immunosorbent assay to detect TNF-alpha. Corneal epithelial cell damage was examined with a slit-lamp biomicroscope and scored by the van Bijsterveld scoring system. RESULTS: TNF-alpha was detected in the tears of patients with SS at concentrations ranging from 8 to 1,500 pg/ml in 33 of the 60 eyes of the patients. In contrast, TNF-alpha was not detected in the tears of normal controls. The concentration of TNF-alpha was significantly higher in the tear samples from patients whose clinical scores were two or three points compared with patients whose scores were zero or one point (Mann-Whitney U test: p < 0.03). CONCLUSIONS: TNF-alpha was detected in the tears of patients with SS, and tear TNF-alpha levels showed a significant correlation with the grade of corneal epithelial damage, suggesting that TNF-alpha is a potent mediator in keratoconjunctivitis sicca.

Enzyme-Linked Immunosorbent Assay↗

PubMed2004

[The relationship between Staphylococcus aureus and atopic keratoconjunctivitis].

PURPOSE: To investigate the role of Staphylococcus aureus in patients with vernal keratoconjunctivitis (VKC) and atopic keratoconjunctivitis (AKC) complicated by atopic dermatitis (AD). SUBJECTS AND METHODS: Microbiological culture from the conjunctival sac of acute exacerbated patients with VKC and AKC complicated by AD was performed. The subjects were 29 patients (31 eyes) with VKC and AKC who showed acute exacerbated clinical symptoms of conjunctivitis. Antigen specific IgE antibodies to staphylococcal enterotoxin A (SEA) and staphylococcal enterotoxin B (SEB) in serum were also examined. In this study, the patients were divided into two groups: An AD group consisting of 22 patients with VKC and AKC complicated by AD, and a control group consisting of 8 patients with allergic conjunctivitis without AD. We also examined SEA/SEB specific IgE antibody in tears from the 5 patients who underwent bacteriological examinations of the conjunctival sac at the same time. RESULTS: Twenty-seven out of 31 eyes were gram-positive in the bacteriological culture from the conjunctival sac (87.1%). Staphylococcus aureus was detected in 21 out of the 27 eyes (77.8%). In the AD group, 2 of the 22 cases were gram-positive for serum SEA specific IgE antibodies, 2 cases of the 22 cases were gram-positive for SEB specific IgE antibodies, and 9 cases were gram-positive for both SEA and SEB specific IgE antibodies. Serum SEA and SEB specific IgE antibodies were all gram-negative in the control group. Either SEA or SEB specific IgE antibody in tears was detected in all of the above 5 patients who underwent bacteriological examinations of the conjunctival sac, and 4 of the 5 were gram-positive. Staphylococcus aureus was isolated in 3 out of the 5 patients, and 1 case was gram-negative. CONCLUSION: Staphylococcus is one of the exacerbating factors in VKC and AKC. It is important to evaluate both bacteriological examinations of the conjunctival sac and SEA/SEB specific IgE in tears.

Conjunctiva↗

PubMed2003

A novel grading method for superficial punctate keratopathy magnitude and its correlation with corneal epithelial permeability.

OBJECTIVES: To propose a novel grading method for superficial punctate keratopathy (SPK) magnitude and to examine the quantitativeness of the method. METHODS: In 351 eyes diagnosed as having dry eye syndrome, SPK was graded as follows. After fluorescein staining, the total sum of the area of SPK was graded from A0 through A3, and the density was graded from D0 through D3. The grading was represented as the combination of the area and density grades. The correlation between the SPK grade and the corneal epithelial permeability to fluorescence, measured using an anterior fluorophotometer, was analyzed in the 351 eyes. MAIN OUTCOME MEASURES: The correlation between the fluorescein concentration in the cornea and the sum of the area and density grades. RESULTS: The higher the density and area grades, the higher the fluorescein concentration, except for A3D2. There was a clear exponential correlation between the sum of the area and density grades and the fluorescein concentration in the cornea, measured using an anterior fluorophotometer. The regression curve was y = 61.2e(0.59x), where y is the fluorescein concentration (nanograms per milliliter), and x is the sum of the area and density grades (P<.001). CONCLUSIONS: The SPK grade is positively correlated with the corneal epithelial permeability to fluorescence, measured using an anterior fluorophotometer. Our results indicate that this grading method can be clinically useful.

Corneal Diseases↗

PubMed2003

[Clinical evaluation of measurement method for antigen specific IgE in tears of patients with allergic conjunctival disease].

PURPOSE: To investigate a method for determining antigen specific IgE antibodies in tears of patients with allergic keratoconjunctival disease. SUBJECTS AND METHODS: Antigen specific IgE antibodies to Japanese cedar pollen or Housedust-Mites in tears of patients with allergic conjunctival diseases were examined. Both eyes in each patient were examined. The right eye was examined in normal healthy volunteers as a control. The number of eyes examined for IgE antibodies to Japanese ceder pollen was 32 eyes with seasonal allergic conjunctivitis(SAC), 12 eyes with perennial allergic conjunctivitis(PAC), 12 eyes with atopic keratoconjunctivitis (AKC), 4 eyes with vernal keratoconjunctivitis (VKC), and 12 eyes of the controls. The number of eyes examined for IgE antibodies to Housedust-Mites was 32 eyes with SAC, 20 eyes with PAC, 30 eyes with AKC, 22 eyes with VKC, and 10 eyes with the control. Tears were sampled by the method of Schirmer Test-1. Sampled tears were eluded with 200 microliters of phosphate buffered solution(pH 7.2, 0.05 M) and analyzed by the AlaSTAT-IMMULYZE method. Recovery rate of absorbance by filter paper for IgE antibodies in tears was determined by ratio to the standard sample of serum solution containing Housedust-Mite specific IgE antibodies with known concentration. RESULTS: Recovery rate of filter paper for IgE antibodies in tears was 83.6%. IgE antibodies in tears to Japanese cedar pollen were detected in 11 of 32 eyes of SAC. In other subjects, IgE antibodies were under the limit of the detection concentration except one eye of a patient with AKC. In the positive cases of Japanese cedar pollen specific IgE antibodies, there was no significant difference in the concentration between the right and the left eyes. IgE antibodies to Housedust-Mites were significantly higher not only in incidence but also in concentration in 18 of 22 eyes with VKC than in that in other in 8 of 20 eyes with PAC and 12 of 30 eyes with AKC. CONCLUSION: The method for determining antigen specific IgE antibodies in tears is clinically useful to diagnose and to investigate pathophysiology of allergic conjunctival diseases.

Adolescent↗

PubMed2002

CD40 expression in normal human cornea and regulation of CD40 in cultured human corneal epithelial and stromal cells.

PURPOSE: To determine whether CD40-CD40 ligand (CD40L) interaction plays a role in corneal inflammatory responses, the expression of CD40 and CD40L on normal human cornea was investigated. In addition, using cultured human corneal epithelial (HCE) and human corneal stromal (HCS) cells, the regulation of CD40 expression in human corneal cells investigated, including that induced by proinflammatory cytokines such as interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha. METHODS: Frozen optimal cutting temperature (OCT) compound-embedded sections of corneal tissues obtained from 18 normal human corneas were examined by an immunoperoxidase staining technique with anti-CD40 and anti-CD40L monoclonal antibodies (mAbs). Also, cultured HCE and HCS cells, with IFN-gamma (250-1000 U/mL) or TNF-alpha (500-4000 U/mL) treatment for 1 to 4 days and with no treatment, were stained by the immunofluorescence technique with mAbs and analyzed by flow cytometry. RESULTS. The area of positive staining for CD40 showed a topographical difference. The limbal epithelial cells were predominantly positive for CD40. Positive staining was also found to a lesser extent on the cells in the basal layer of peripheral corneal epithelium. Epithelial cells of the central cornea showed no immunoreactivity for CD40. Corneal stromal cells were negative for CD40 in most of the donor tissues (positive: 5 of the 18 corneas). Endothelial cells were distinctly negative for CD40. Cultured HCE cells were also positive but decreased in positive cell number with lengthening culture period. None or less than 5% of the cultured HCS cells were CD40 positive. IFN-gamma enhanced CD40 expression on both cell types. In contrast, TNF-alpha enhanced CD40 on HCE but not on HCS cells. No component cells of normal human cornea or cultured HCE and HCS cells showed immunoreactivity for CD40L. CONCLUSIONS: In the human cornea, CD40 is expressed predominantly on limbal epithelial cells and also on cultured HCE cells with high proliferative potential. In addition, the expression of CD40 is induced on cultured HCE and HCS cells differentially by proinflammatory cytokines, such as IFN-gamma and TNF-alpha.

Antibodies, Monoclonal↗

PubMed2002

[Histological study of conjunctiva-associated lymphoid tissue in mice].

PURPOSE: To investigate conjunctiva-associated lymphoid tissue (CALT) in mouse conjunctiva. METHOD: We examined normal mice, ranging in age from 4 to 6 weeks, to investigate the presence of follicular tissue histologically. Next, we treated mice by topical instillation of ovalbumin and cholera toxin B, and then examined conjunctival follicles by immunohistological methods. RESULTS: Follicular tissue was present at the nictitating membrane. Both size and amount of follicular tissue was increased by the topical ovalbumin treatment. Immunohistological examination revealed CD4, CD8, and S-100 protein positive cells in follicular tissue. The epithelial layer, corresponding to follicular tissue, had intraepithelial pockets and CD4 positive cells in the pockets. CONCLUSION: It was concluded that the follicular tissue at the nictitating membrane is CALT in mice.

Animals↗

PubMed2002

[Clinical study on surgical outcome of penetrating keratoplasty for herpetic leukoma].

PURPOSE: To evaluate prognostic factors, rejection, and recurrence of herpetic keratitis, in keratoplasty for herpetic leukoma. SUBJECTS AND METHODS: We compared surgical outcome between patients who underwent keratoplasty for herpetic leukoma and those who received it for non-herpetic disorders. We compared the eyes of 24 patients in the herpetic group with 17 eyes of 20 patients with non-herpetic corneal disorders, i.e., the non-herpetic group. RESULTS: The rejection rate was 45.8% for the herpetic group and 5.00% for the non-herpetic group, showing significant difference(p < 0.05). Transparent grafts were obtained for 75.0% of the herpetic group and 85.0% of the non-herpetic group, giving no significant difference. In the herpetic group, graft transparency in eyes with recurrences were 94.1%(p < 0.05). Graft rejection occurred mostly within 1 year postoperatively. RecuRrent herpetic keratitis tended to occur within 3 years and it was later than graft rejection. CONCLUSION: In patients who underwent keratoplasty for herpetic leukoma, steroid therapy is important for 1 postoperative year to suppress graft rejection, but after that, steroid treatment should be carefully considered to minimize the risk of recurrences of herpetic keratitis.

Aged↗

PubMedDate not supplied

Gelatinase expression in ocular surface disorders.

PURPOSE: To determine the expression of gelatinase A [matrix metalloproteinase (MMP)-2] and gelatinase B (MMP-9) in ocular surface disorders, we carried out examinations of the eyes of patients with such disorders. METHODS: The cases consisted of a normal group comprising three eyes as the control and a patient group comprising six eyes of patients with ocular surface disorders. These groups underwent the following examinations. (1) Gelatin zymography: Tear samples taken by using the Schirmer I test were eluted and analyzed by gelatin zymography. (2) Immunohistochemical study: By using the alkaline phosphate-anti-alkaline phosphatase or avidin-biotin complex method, corneal or conjunctival tissue was examined for the presence of MMP-2, MMP-9, and membrane type 1 (MT1)-MMP, which activates MMP-2. (3) Reverse transcription-polymerase chain reaction (RT-PCR): Total RNA was eluted from the specimens, and the expression patterns of MMP-2, MMP-9, and MT1-MMP mRNA were analyzed. RESULTS: (1) Gelatin zymography: Only the proforms of MMP-2 and MMP-9 were detected in the normal group. Both active MMP-2 and active MMP-9 were detected in all studied cases in the patient group. (2) Immunohistochemical study: In the normal group, only MMP-2-positive cells were present in conjunctival epithelium. In the patient group, cells positive for MMP-2, MMP-9 and MT1-MMP were present in the epithelium of the conjunctiva or cornea except in one thermal burn case. (3) RT-PCR: In the normal group, only MMP-2 expression was observed. MMP-2, MMP-9, and MT1-MMP mRNA expression was observed in three cases in the patient group. CONCLUSIONS: MMP-2, MMP-9, and MT1-MMP, which activates MMP-2, were expressed in the patients with various ocular surface disorders. In ocular surface disorders, therefore, the expression of gelatinases is upgraded.

Conjunctival Diseases↗

PubMedDate not supplied

Histological study on mast cells in conjunctiva of NC/Nga mice.

PURPOSE: To investigate the pathological findings in conjunctiva of NC/Nga mice, which develop atopic dermatitis spontaneously, we focused our study on the density of mast cells as determined by histological methods. METHODS: NC/Nga mice were divided into five groups; the 4W-group comprised four 4-week-old mice without treatment, the 10W-group comprised four 10-week-old mice without treatment, the 16W-group comprised four 16-week-old mice without treatment, the Dx group comprised three 16-week-old mice undergoing topical 0.1% dexamethasone (Dx) ointment treatment, and the FK506 group comprised three 16-week-old mice undergoing topical 0.1% FK506 ointment treatment. For the histological examination, the lids and eyeballs of the mice were removed and fixed with Carnoy's solution and thin sections were made. The Carnoy-fixed specimens were stained with toluidine blue or H&E and examined histologically. Toluidine blue-stained tissue sections were examined for the density per square millimeter of mast cells, which were identified as metachromatic cells in the conjunctival tissue and lid cutaneous tissue by light microscopy. RESULTS: Mast cell density in the conjunctiva was 8.6 +/- 8.2 cells/mm(2) in the 4W-group, 29.2 +/- 22.0 cells/mm(2) in the 10W-group, 41.0 +/- 21.1 cells/mm(2) in the 16W-group, 22.7 +/- 17.1 cells/mm(2) in the Dx group, and 33.6 +/- 27.7 cells/mm(2) in the FK506 group. Mast cell density increased significantly with age among the 4W-, 10W-, and 16W-groups ( P < 0.001). The mast cell density of lid cutaneous tissue was higher than the conjunctival mast cell density. Mast cell density was significantly higher in the 16W-group than in the Dx group, but not significantly higher than in the FK506 group. CONCLUSIONS: An increase in mast cell density with age indicates that NC/Nga mice develop atopic keratoconjunctivitis-like signs. Dexamethasone ointment has a suppressive effect on the increase in mast cell density.

Aging↗

PubMedDate not supplied

Expression of alpha- and beta-defensins in human ocular surface tissue.

PURPOSE: Defensins are antimicrobial peptides and are classified into three groups, alpha-, beta- and theta-defensins. We investigated the localization of human neutrophil peptides (HNP) 1-3, which are members of the alpha-defensin subfamily, and the expression of human beta-defensin 1 (hBD-1) and hBD-2 in the ocular surface tissues. METHODS: Corneal or conjunctival tissues were obtained from 17 patients undergoing surgery. The reasons for surgery were thermal burns (two patients), acid burn (one patient), corneal perforation (one patient), corneal abscess (one patient), and pterygium (two patients). As a control, tissue samples were collected from ten patients undergoing cataract surgery. The samples were examined for HNP 1-3 by immunohistochemical analysis, and for hBD-1 and hBD-2 by the reverse transcriptase-polymerase chain reaction (RT-PCR) method. RESULTS: Results of staining for HNP 1-3 were positive in all of the six tested samples. The RT-PCR method detected hBD-1 mRNA in all samples. However, hBD-2 was detected in only 7 of the 17 samples. In eight of the ten normal conjunctiva samples and in the two samples from conjunctiva with thermal burn or pterygium, hBD-2 mRNA was not detected. CONCLUSIONS: We concluded that HNP 1-3 and hBD-1 are constitutively expressed in the ocular surface. However, hBD-2 may or may not be expressed, depending on the stage of ocular surface diseases.

Adult↗

PubMedDate not supplied

Comparison of topical dexamethasone and topical FK506 treatment for the experimental allergic conjunctivitis model in BALB/c mice.

PURPOSE: To evaluate the efficacy of topical dexamethasone and topical FK506 treatment on allergic inflammation in conjunctiva, we performed a comparative study using a mouse experimental allergic conjunctivitis model. METHODS: The experimental allergic conjunctivitis model was developed by ovalbumin (OVA) instillation after OVA sensitization by intraperitoneal application of a mixed solution of OVA and aluminum hydroxide. The Balb/c mice of the experimental allergic conjunctivitis model were divided into three groups, depending on whether or not they received topical treatment and which type of treatment they received. The allergy group comprised six mice without topical treatment; the dexamethasone (Dx) group comprised six mice receiving topical 0.1% dexamethasone ointment treatment; and the FK506 (FK) group comprised six mice receiving topical 0.1% FK506 ointment treatment. The negative control group comprised six mice with neither sensitization nor topical treatment. For the histological examination, frozen sections of the eyelids, eyeballs, and lacrimal glands were stained with acid Giemsa stain or, in the immunohistochemical method, with alkaline phosphatase-antialkaline phosphatase (APAAP), for CD4. Densities of eosinophils and CD4-positive lymphocytes in the conjunctival tissue were counted. Productive C epsilon gene expression in the conjunctival tissue, including the lacrimal gland, was also evaluated by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: In the histological study, the cell density of eosinophils infiltrating into subconjunctival tissues was 66.3 +/- 34.7 cells/mm(2) (mean +/- SD) for the allergy group, 2.5 +/- 4.2 cells/mm(2) for the Dx group, 4.2 +/- 8.6 cells/mm(2) for the FK group, and 3.7 +/- 7.6 cells/mm(2) for the negative control group. The cell density of CD4-positive cells infiltrating into subconjuctival tissues was 145.9 +/- 66.4 cells/mm(2) for the allergy group, 10.0 +/- 12.9 cells/mm(2) for the Dx group, 10.1 +/- 15.5 cells/mm(2) for the FK group, and 21.7 +/- 17.8 cells/mm(2) for the negative control group. The allergy group showed significant differences in the density of eosinophils and CD4-positive cells compared with those of the Dx (P < 0.0001) and FK (P < 0.0001) groups. In the RT-PCR study, the expression of productive C epsilon gene was positive in the Dx group, but negative in the allergy group, the FK group, and the negative control group. CONCLUSIONS: Topical Dx and FK506 treatments were both efficacious in suppressing eosinophil and lymphocyte infiltration into subconjunctival tissue in the mouse experimental allergic conjunctivitis model. However, the influence of their IgE productive mechanisms seemed to differ, causing different efficacy of productive C epsilon gene expression in the conjunctival tissue.

Administration, Topical↗