Effects of astaxanthin on antioxidation in human aqueous humor.

We evaluated the antioxidative effects of astaxanthin through the changes in superoxide scavenging activity, levels of hydrogen peroxide and total hydroperoxides in human aqueous humor. The study subjects were 35 patients who underwent bilateral cataract surgery on one side before and the other side after intake of astaxanthin (6 mg/day for 2 weeks). Their aqueous humor was taken during the surgery and subjected to measurements of the three parameters. After astaxanthin intake, the superoxide scavenging activity was significantly (p<0.05) elevated, while the level of total hydroperoxides was significantly (p<0.05) lowered. There was a significant negative correlation between the superoxide scavenging activity and the level of total hydroperoxides (r = -0.485, p<0.01), but no correlations between the hydrogen peroxide level and the other two parameters. Astaxanthin intake clearly enhanced the superoxide scavenging activity and suppressed the total hydroperoxides production in human aqueous humor, indicating the possibility that astaxanthin has suppressive effects on various oxidative stress-related diseases.

The role of nick formation in delayed quinacrine mustard fluorescence in the C-heterochromatin of Apodemus argenteus.

Chromosomes stained with fluorochromes, including quinacrine mustard (QM), emit the brightest fluorescence immediately after exposure to excitation light, and the fluorescence gradually fades with an increase in exposure time. However, in the QM-stained chromosomes of the small Japanese field mouse Apodemus argenteus, most C-heterochromatic regions emit weak fluorescence immediately after exposure to blue light, and they become brightly fluorescent by prolonged exposure (delayed QM fluorescence). We proposed recently that the delayed QM fluorescence is somehow related to nicks produced in C-heterochromatic DNA by blue light irradiation. To test this possibility, we examined the chromosomal distribution of nicks by in-situ nick translation and changes, if any, in the QM fluorescence pattern after methylene blue (MB) -mediated photooxidation, which is considered to induce nicks in chromosomal DNA. It was found that C-heterochromatic regions fluoresced brightly without any delay after exposure to blue light, and that nicks increased considerably in the same regions after the MB-mediated photooxidation. It seems, therefore, that photooxidation and strand breaks in DNA (including nicks) are responsible for the induction of delayed QM fluorescence. Trypsin digestion, on the other hand, abolished delayed QM fluorescence. Thus, not only DNA but also chromosomal protein(s) are involved in the unusual sequence of QM fluorescence patterns in A. argenteus.

Genotoxic assessment of small mammals at an illegal dumpsite at the Aomori-Iwate prefectural boundary.

In 2003, we examined the chromosomes of grass voles at an illegal dumpsite at the Aomori-lwate prefectural boundary. In subsequent years, from 2003-2006, we surveyed the chromosomes of four species of small mammals, namely, the Japanese grass vole (Microtus montebelli), the large Japanese field mouse (Apodemus speciosus), the small Japanese field mouse (A. argenteus), and the greater Japanese shrew mole (Urotrichus talpoides). Each annual survey revealed, both on a yearly basis and during the entire period in question, that the frequencies of breaks and gaps in chromosomes of M. montebelli were significantly higher at the dumpsite than on the outskirts and in controls, suggesting that grass voles at the dumpsite have been subject to continuous genotoxic effects since the establishment of the dumpsite. We also ascertained that grass voles are much more susceptible to chromosomal damage than field mice and shrew moles, which had very low levels of chromosomal aberrations at the dumpsite, on the outskirts of the dumpsite, and in controls. Our four-year survey revealed two variants of M. montebelli from the dumpsite with M6 fission (2n=31), two variants of A. speciosus from the outskirts with XO monosomy (2n=47, XO), and a variant of A. speciosus from the dumpsite with situs inversus. Our analysis confirms our previously proposed hypothesis that M. montebelli might be useful as an indicator species for genotoxic assessment of below-ground pollution by industrial waste at illegal dumpsites.

Chromosomal aberrations in Japanese grass voles in and around an illegal dumpsite at the Aomori-Iwate prefectural boundary.

Bone marrow chromosomes of thirty specimens of the Japanese grass vole, Microtus montebelli (2n=30), which had been caught on and near an illegal dumpsite at the Aomori-Iwate prefectural boundary, were analyzed and compared with those of fifteen grass voles from non-polluted areas as part of an effort to assess genotoxic influences on grass voles in the dumpsite area. Fifty metaphases per specimen were examined with particular attention to numerical and structural aberrations. Two specimens from the dumpsite had 2n=31, which was confirmed by G-banding analysis to have been caused by centric fission of M6 homologs, while control specimens had no such abnormality. In specimens from the polluted area, the mean number of chromosomal aberrations (breaks and/or gaps) per 50 metaphases per specimen was 2.57+/-0.41, which was significantly higher than that (0.80+/-0.14; P<0.01) in control specimens. Chromosomal aberrations were randomly distributed on chromosomes, with frequencies being proportional to the relative lengths of chromosomes. Our findings suggest that grass voles at and around the dumpsite have been seriously damaged at the chromosomal level and, moreover, that M. montebelli might be useful as an indicator species for genotoxic assessment of below-ground pollution by industrial waste at illegal dumpsites.

[Oxidative stress in ocular disease].

The definition of oxidative stress implies increased oxidant production in animal cells characterized by the release of free radicals, resulting in cellular degeneration. The imbalance between excess free radical production and the antioxidant defense causes cellular damage resulting in lipid peroxidation. Oxidative stress is involved in many ocular diseases such as age-related macular degeneration, retinopathy of prematurity, retinal light damage, and cataract. Reactive oxygen species are involved in this process. The pathogenesis of age-related macular degeneration is largely unknown. Excessive light and iron may enhance the progression of this disease. In in vitro study of the ciliary body, gamma irradiation inhibits TPR53BP2 expression associated with apoptotic cell death, and increased BCL2 is evident just after gamma irradiation. Exposure to ultraviolet light has been postulated as a cause of age-related macular degeneration (AMD), perhaps through damage to the retinal pigment epithelium. It seems logical, therefore, to replace the aging, yellowing lens with a blue light-absorbing yellow intraocular lens (IOL) in cataract surgery. The issue of whether cataract surgery is a risk factor for the development or progression of AMD remains controversial. In vivo studies suggest that lipid peroxidation decreases in the vitreous and retina after cataract surgery with or without intraocular lens implantation.

The delayed quinacrine mustard fluorescence from the C-blocks of Apodemus argenteus is due to the introduction of nicks into the DNA.

"Delayed QM-fluorescence" refers to the unusual kinetics of fluorescence from most of the C-heterochromatic regions of the chromosomes of the small Japanese field mouse Apodemus argenteus. When stained with quinacrine mustard (QM-stained), these C-heterochromatic regions emit weak fluorescence immediately after exposure to blue light (BL); they emit bright fluorescence within a few minutes; and the intensity of the fluorescence gradually decreases after maximum fluorescence has been recorded. To elucidate the mechanism of this phenomenon, we used acridine orange staining (AO-staining) and a modified version of the in situ nick-translation method. Focusing on the large C-heterochromatic region (C-block) of the X chromosome, we noted that AO-stained C-blocks emitted greenish fluorescence, while QM-stained and BL-exposed (QM-BL-processed) C-blocks emitted reddish fluorescence upon AO-staining after removal of QM. These findings suggested that the C-block DNA of A. argenteus might undergo a structural change, such as strand breaks, during QM-BL processing. Application of the modified in situ nick-translation method revealed the generation of an appreciable number of nicks in the C-block DNA by QM-BL processing. No such nick formation was observed in the C-blocks of three other mammalian species: Apodemus peninsulae, Microtus montebelli, and Urotrichus talpoides. Our findings support the hypothesis that nick formation due to exposure to BL might play a primary role in inducing delayed QM-fluorescence in the C-blocks of A. argenteus. On the basis of the present and earlier findings, we propose a probable mechanism for delayed QM-fluorescence in A. argenteus chromosomes.

Visual acuity prognosis after anterior chamber air replacement to prevent pseudo-anterior chamber formation after deep lamellar keratoplasty.

PURPOSE: To investigate the prognosis of patients who received anterior chamber air replacement after deep lamellar keratoplasty (DLKP) during the study period, January 1995 to April 2000. METHODS: The records were studied of 47 patients (54 eyes) (60.6 +/- 21.3 years of age) who underwent DLKP at Dokkyo University Hospital. Visual acuity and endothelial cell loss were assessed in patients (1) with and without Descemet's membrane perforation; (2) with and without the use of anterior chamber air replacement, and for different durations of air replacement; and (3) in the presence or absence of a pseudo-anterior chamber, and in relation to its duration if present. RESULTS: No significant differences in relation to the above three items were found in endothelial cell loss in study years 1 to 5. Average best visual acuity was 0.61 in perforated eyes, 0.54 in unperforated eyes, 0.54 in eyes that received air replacement, and 0.57 in eyes that did not. The average best visual acuity was 0.38 in eyes with a pseudo-anterior chamber and 0.68 in eyes without one. There was a significant correlation between the duration of the pseudo-anterior chamber and loss of visual acuity. CONCLUSIONS: The prolongation of a pseudo-anterior chamber eventually impairs visual acuity, whereas anterior chamber air replacement, used to prevent the development of a pseudo-anterior chamber, causes minimal endothelial cell damage. Anterior chamber air replacement, therefore, is an effective technique by which to prevent the development of a pseudo-anterior chamber.

p27kip1 siRNA induces proliferation in corneal endothelial cells from young but not older donors.

PURPOSE: To determine whether small interfering (si)RNA downregulation of the cyclin-dependent kinase inhibitor p27kip1 overcomes G(1)-phase arrest and promotes cell-cycle progression in human corneal endothelial cells (HCECs) from young (<30 years old) and older (>60 years old) donors. METHODS: Transfection of siRNA was confirmed by incubating confluent cultures of HCECs with FITC-labeled nonsilencing siRNA. Confluent cultures were transfected for 48 hours with p27kip1 siRNA (2.5, 5, 25, or 100 nM) or nonsilencing siRNA, with a lipid transfection reagent. As a comparison, cultures were also transfected for 48 hours with p27kip1 antisense (AS) or missense (MS) oligonucleotides (oligo). At various times after transfection, cells were fixed for immunocytochemical localization of p27kip1 or extracted for Western blot analysis to assess relative p27kip1 protein levels. Cultures were also prepared for ZO-1 immunolocalization, to assess the effect of transfection on the morphology of the monolayer. The number of cells was counted at 0, 48, 96, 144, and 192 hours after incubation, and a cell-viability assay was performed. RESULTS: A dose-dependent decrease in p27kip1 protein level was observed in Western blot analysis, and nuclear staining for p27kip1 was greatly reduced in HCECs incubated with p27kip1 siRNA. No change in p27kip1 levels or in nuclear staining was observed in the nonsilencing control. p27kip1 siRNA (25 nM) appeared to be quantitatively more efficient than antisense oligonucleotide (500 nM) in reducing p27kip1 protein levels. Viability was less affected by siRNA treatment than by AS oligo transfection. ZO-1 staining showed no effect on morphology of the monolayer. The number of HCECs from young donors (<30 years old) transfected with p27kip1 siRNA increased up to 144 hours after incubation, whereas no change in the number of cells was observed in HCECs transfected with nonsilencing siRNA. In contrast to the results from young donors, no change in the number of cells was observed at any time point tested in HCECs from older donors (>60 years old) after p27kip1 siRNA transfection. CONCLUSIONS: Transfection of p27kip1 siRNA was sufficient to promote proliferation in confluent cultures of HCECs from younger, but not older donors. These results suggest that inhibition of proliferation in older donors is regulated by other mechanisms in addition to p27kip1.

Active oxygen processing for acrylic intraocular lenses to prevent posterior capsule opacification.

PURPOSE: To evaluate active oxygen processing on the surface of acrylic intraocular lenses (IOLs) to prevent secondary posterior capsule opacification (PCO). SETTING: Department of Ophthalmology, Dokkyo Medical University School of Medicine, Mibu City, Tochigi, Japan. METHODS: Acrylic IOLs were prepared, and ultraviolet (UV)/ozone (UV/O3) or argon plasma was irradiated to the surface of the IOLs. Elemental analysis (electron spectroscopy for chemical analysis [ESCA]) of the IOL surfaces was performed to confirm surface modification. Changes produced by UV/O3 or argon plasma treatment were examined for fibronectin and lens epithelial cell (LEC) adhesion. To evaluate the PCO prevention by treated IOLs, 8-week-old albino rabbits were used. The rabbit eyes randomly had phacoemulcification and implantation of 3 different IOLs: the UV/O3-treated IOLs, argon plasma-treated IOLs, and the control IOLs. After 2 weeks, the rabbits were killed and their globes were dissected and fixed using formaldehyde 10%. The PCO was observed using light microscopy (DX51, ORIMPUS) after hematoxylin and eosin staining. RESULTS: Comparison of IOL surface composition by ESCA showed an increase in nitrogen content and hydroxyl substitute and carboxyl substitute groups on surfaces of treated IOLs. The fibronectin adhesion and the LEC adhesion on the UV/O3-treated and argon plasma-treated samples were increased. In the untreated group, there was statistically significant inhibition of PCO formation in the UV/O3-treated and argon plasma-treated groups. CONCLUSION: Active oxygen processing and argon plasma irradiation on the surface of IOLs was effective in preventing secondary PCO after cataract surgery.

[Effects of calcium on lens epithelial cells in rabbits].

PURPOSE: The action of lens epithelial cells (LECs) is important for cataract and posterior subcapsular cataract after cataract surgery. In this study, we analyzed the effects of calcium on the characteristics of LECs. METHODS: The LECs were collected using albino rabbits and incubated in minimum essential medium [MEM, Introgen Corp. (12% fetal bovine serum: FBS)] (37 degrees C, 5 % CO2) for a week to induce their proliferation. Cell culture dishes (35 mm) were prepared and 7 mm cylindrical pipes were placed in them. After that, around 10,000 cultured LECs were placed in the pipes and incubated. After 2 hours incubation, the pipes were removed and various doses of MEM (0, 2, 10 and 20 mM) replaced the calcium. Proliferation and shapes of LECs were observed using a confocal microscope and immunohistological analysis [alpha-smooth muscle actin (alpha-SMA) and bromodeoxyuridine (BrdU)]. The LECs were incubated with collagen gel and different calcium doses (0, 2, 10 and 20 mM) of MEM to calculate the contraction rate. RESULTS: It was observed that the LECs changed to fibroblast-like cells at high doses of calcium using a confocal microscope. Histological studies showed that the BrdU positive cells were increased by using 10 and 20 mM calcium MEM, but the positive cells were decreased by using 0 and 2 mM calcium MEM. Increase of alpha-SMA stained cells was recognized when using 0, 10 and 20 mM calcium MEM. The contraction rate of collagen gel was increased by using the 10 and 20 mM calcium MEM. CONCLUSION: The changes of calcium concentration might be an important factor for the development of cataract, posterior subcapsular opacification, and contraction of the lens capsule after cataract surgery.

[Protective effect on the corneal endothelium and remaining effect at the anterior chamber for three different kinds of viscoelastic devices].

PURPOSE: The present study was performed to evaluate the corneal endothelium protection and anterior chamber stagnation abilities of three different types of viscoelastic substances (Healon, Viscoat, HealonV). METHODS: Viscoelastic substances were selected at random for 120 eyes with cataracts, and the postoperative reduction rates of the corneal endothelium cells were compared. The residual viscoelastic substances after filling of the anterior chamber of pig eyes and aspiration with a handpiece were measured by an anterior eye segment image analysis system. The same procedures were performed in rabbit eyes and the residual levels of viscoelastic substances on the corneal endothelium were photographed histologically. RESULTS: The reduction rate of endothelium corneal cells tended to decrease with Viscoat three months after surgery. The results obtained with the anterior eye segment image analysis system showed that the residual level in the anterior chamber was higher with Healon. Histological analyses demonstrated residual Viscoat at the center of the corneal endothelium after perfusion. CONCLUSION: HealonV was superior in terms of spatial retention and Viscoat had corneal endothelium protection potential.

The effects of drug delivery via hydrophilic acrylic (hydrogel) intraocular lens systems on the epithelial cells in culture.

BACKGROUND AND OBJECTIVE: Secondary posterior subcapsular opacification is still among the most important complications after phacoemulsification. MATERIALS AND METHODS: This study was designed to assess the inhibitory effects of drugs delivered via hydrophilic acrylic (hydrogel) intraocular lens (IOL) systems in vitro. Lens epithelial cells were collected from albino rabbits. The following seven groups of hydrogel IOLs were prepared: untreated IOLs and IOLs infiltrated with diclofenac sodium, tranilast, mitomycin C, colchicines, 5-fluorouracil, and ethylenediaminetetraacetic acid. The IOLs were fixed to a Cell Culture Insert; they were then bathed and incubated in minimum essential medium containing cultured lens epithelial cells. Subsequently, a comparative analysis of the cells adhering to the collagen membrane and the lens surfaces was conducted. RESULTS: Adhesion of lens epithelial cells to the lens surfaces and the collagen membrane was observed in the control group. However, only slight cellular adhesion was found on the surfaces of the IOLs and on the collagen membrane in the treated IOL groups. CONCLUSION: Use of hydrogel IOLs infiltrated with drugs was associated with inhibition of posterior subcapsular opacification in vitro.

p27kip1 Antisense-induced proliferative activity of rat corneal endothelial cells.

PURPOSE: To determine whether antisense downregulation of p27(kip1) will overcome G(1)-phase arrest and promote cell cycle progression in rat corneal endothelial cells (CECs). METHODS: Confluent cultures of rat CECs were incubated for 24 hours in the presence of p27(kip1) antisense (AS) oligonucleotides (oligoS) using nonliposomal lipid transfection. Control cultures were incubated under one of the following conditions: no oligos or lipid-containing buffer, lipid-containing buffer alone, or lipid-containing buffer plus missense (MS) p27(kip1) oligo. Viability was tested by a cell-viability assay after 0, 24, 48, and 72 hours. After postincubation for 0, 24, 48, or 72 hours, cultures were fixed and immunostained for p27(kip1), to test for downregulation, or for Ki67 or BrdU, to detect actively cycling cells. Western blot and immunocytochemistry (ICC) studies were conducted to determine the effect of p27(kip1) antisense treatment on the relative protein level and subcellular localization of several cell cycle proteins, including cyclin-D1, -E, -A, and -B1; CDK2 and -4; p21(cip1); and p15(INK4b). Proliferation was determined by direct counting of propidium iodide (PI) or 4',6'-diamino-2-phenylindole (DAPI)-stained cells. RESULTS: Viability was not significantly affected by lipid-based oligo transfection for up to 48 hours, after which a decline was noted. The protein level of p27(kip1) was reduced after AS transfection in a time-dependent manner. Nuclear staining for p27(kip1) was greatly reduced in CECs incubated with AS oligo. No change in p27(kip1) levels was observed in controls at any time point tested. p27(kip1) AS oligo transfection increased cyclin-D1, -E, -A, and -B1 protein levels, and all cyclins were localized to the nucleus. No changes in protein level were observed for CDK2, CDK4, p21(cip1), or p15(INK4B). A time-dependent increase in the relative number of Ki67- and BrdU-positive cells was noted in CECs incubated with AS oligo. In contrast, no to few Ki67- or BrdU-positive cells were observed in CECs incubated with MS oligo or the buffer-treated control cells. The percentage increase in the number of cells transfected with AS oligo increased with time, compared with that of cells transfected with MS oligo. CONCLUSIONS: Treatment with p27(kip1) antisense oligonucleotides followed by postincubation in 10% FBS lowers endogenous p27(kip1) protein levels and promotes proliferation in confluent cultures of rat CECs.

Analysis of cytoskeletal proteins in posterior capsule opacification after implantation of acrylic and hydrogel intraocular lenses.

PURPOSE: To analyze selected lens cytoskeletal proteins in posterior capsule opacification (PCO) 2 weeks after intraocular lens (IOL) implantation in rabbits. SETTING: Department of Ophthalmology, Dokkyo University School of Medicine, Tochigi, Japan. METHOD: Eight 10-week-old albino rabbits were prepared and anesthetized for phacoemulsification and aspiration of the crystalline lens and implantation of an acrylic or a hydrogel IOL. Two weeks postoperatively, the rabbits were killed and the IOLs removed for immunohistochemistry. Deparaffinized tissue sections were processed with antibodies against alpha-smooth muscle actin (alpha-SMA) and beta-crystallin to observe the types of PCO with the 2 IOL types. The proteins in the PCO tissue and the normal lens were homogenized, centrifuged, and analyzed using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) densitometric analysis and Western immunoblotting for actin and vimentin. RESULTS: Immunohistochemistry demonstrated a fibroblastic cell type expressing alpha-SMA and partial regeneration of epithelial cells, resulting in a lenticular structure that stained irregularly for beta-crystallin. The immunoreactivity of fibroblast-like cells to beta-crystallin appeared weaker than that of the regenerated lenticular structure. SDS-PAGE showed variability in the content of cytoskeletal proteins in the insoluble fractions of the PCO. Degradation of the cytoskeletal components was greater with the acrylic IOL than with the hydrogel IOL. CONCLUSION: Cytoskeletal proteins expressed during the formation of PCO and IOL implantation may have potential as therapeutic target proteins to improve the biocompatibility of IOLs.

Effects of calcium on human lens epithelial cells in vitro.

PURPOSE: To analyze the effect of calcium on human lens epithelial cells (LECs) in vitro METHODS: Human LECs were obtained from the anterior lens capsule during a cataract operation, and were cultured in minimum essential medium (MEM) containing 12% fetal bovine serum for a week at 37 degrees C, 5% CO(2). The LECs were then isolated with trypsin and placed in culture dishes. To analyze the effects of calcium, the LECs were incubated in MEM with calcium concentrations of 0, 2, 10, or 20 mM. After H&E staining for 72 h, the LECs were analyzed with the Scion imaging program. RESULTS: The LECs proliferated rapidly and their shapes were uniform in 2-mM-calcium MEM. The LECs proliferated more slowly and had irregular shapes in MEM with lower and higher concentrations of calcium. CONCLUSIONS: Calcium affects both the proliferation and the shape of human LECs in culture. Abnormal (hyper- or hypo-) calcium concentrations in the lens and aqueous humor may change the homeostasis of LECs, resulting in cataracts and secondary posterior capsular opacification.

Clinical evaluation of posterior capsule opacification in eyes with different small-incision intraocular lenses.

BACKGROUND AND OBJECTIVE: To present a new method to quantify posterior capsular opacity with an anterior eye segment image analyzer (EAS 1000, NIDEK). PATIENTS AND METHODS: This study was comprised of patients who underwent phacoemulsification intraocular lens (IOL) implantation. Three types of IOLS, acrylic, silicone, and polymethylmethacrylate (PMMA) were allocated to 30 eyes and clinically evaluated. Patients were observed for 3 years postoperatively using an anterior eye segment image analyzer (EAS1000). Opacity was determined by calculating the area of opacity from a retroillumination image. In the retroillumination mode of analysis, the measurement was limited to a 4-mm-diameter region of the pupillary zone to eliminate the influence of anterior capsular opacity. For color map analysis, the threshold level was expressed as the color tone of 0-255 CCT (computer compatible tape). The glare disability was measured to evaluate the three types of IOLs. RESULTS: The color map analysis revealed a time-related increase in the opacity level of patients receiving the PMMA IOL implant. Three years after surgery, the levels were significantly higher in the PMMA group (P < 0.01) compared to the acryl and silicone groups: acryl (17.5 +/- 3.8), silicone (18.0 +/- 6.2%), and PMMA 36.5 +/- 32.9%. CONCLUSION: Quantitative evaluation using an anterior eye segment image analyzer is effective for observing the degree of posterior capsule opacification. The color map analysis using an anterior eye segment image correlated with the visual function revealed that the time-related increase in the opacity level was significant during the third year in patients receiving PMMA IOL implantation.

[Ocular surface temperature of meibomia gland dysfunction patients and the melting point of meibomian gland secretions].

PURPOSE: Since meibomian gland dysfunction (MGD) is thought to be caused by raising of the melting point and degeneration of lipid secretions, the melting point of secretions and the temperature of the ocular surface were investigated. SUBJECTS AND METHODS: Twenty-three patients with MGD [mean age 69.3 +/- 7.2 (mean standard deviation) years) were examined. Seven subjects without MGD (mean age 32.5 +/- 5.8 years) were also observed as controls. METHODS: Solid secretions obtained from MGD patients were heated and the melting point was measured. The weight ratio of lipids and cell components was also determined. RESULTS: Corneal temperature was 32.3 +/- 0.5 (mean +/- standard deviation) degrees C and eyelid temperature was 33.1 +/- 0.5 degrees C in the control subjects. Corneal temperature was 31.1 +/- 0.8 degrees C and eyelid temperature was 32.7 +/- 0.6 degrees C in MGD patients. However, the melting point of the secretions was 34.0 +/- 1.3 degrees C, which was higher than the temperature of the ocular surface and eyelid in both patients and controls. The cell components constituted about 60% of the secretions and the lipid composition, about 40%. CONCLUSION: The cause of MGD is thought to be solid ification of lipids because of elevation of the melting point and the increase in the cell components of the secretions.

[Cytokine mRNA expression in vernal keratoconjunctivitis].

BACKGROUND: Cytokines play an important role in chronic allergic eye disease. OBJECTIVES: We examined how many kinds of cytokines are in giant papillae of vernal keratoconjunctivitis(VKC). METHODS: We resected giant papillae from 6 patients with VKC, and studied mRNA expression of cytokines in them with the reverse transcriptase-polymerase chain reaction(RT-PCR). The cytokines were interleukin(IL)-1 beta, IL-1 receptor antagonist (IL-1 RA), IL-2, IL-4, IL-5, IL-6, IL-8, interferon (INF) gamma, tumor necrosis factor(TNF) alpha, TNF beta, transforming growth factor(TGF) beta 1, and TGF beta 2. RESULTS: In 5 cases, expression of IL-1 beta was positive. Expression of IL-1 RA, IL-5, IL-6, IL-8, TGF beta 1, and TGF beta 2 was positive in 6 cases. TNF alpha was positive in 4 cases. IL-4 and INF gamma were positive in 3 cases. IL-2 and TNF beta were negative in all patients. CONCLUSION: These results suggest that the Th2 cytokines play more important roles than the Th1 cytokines in VKC inflammation.

Relationship between Pentosidine and Pyridinoline Levels in Human Diabetic Cataract Lenses.

The relationship between the levels of two different crosslink compounds, pentosidine and pyridinoline, in human diabetic cataract lenses was investigated to elucidate the pathogenic mechanism of diabetic cataract. Subjects were classified into diabetes mellitus (DM) group and non-DM group according to the presence or absence of DM. The levels of the crosslink compounds were determined using high-performance liquid chromatography and spectrofluorometry after acid hydrolysis. In the non-DM group the pentosidine level was significantly and positively correlated with the pyridinoline level and age. In the DM group the pentosidine level was not significantly correlated with either pyridinoline level or age. Pyridinoline levels and age were not significantly correlated in either group. The increase in crosslink compounds due to glycation and the relationship between the compounds are changed in DM lenses.