The Anti-Inflammatory Effects of Angiogenin in an Endotoxin Induced Uveitis in Rats.

Angiogenin (ANG) is involved in the innate immune system and inflammatory disease. The aim of this study is to evaluate the anti-inflammatory effects of ANG in an endotoxin induced uveitis (EIU) rat model and the pathways involved. EIU rats were treated with balanced salt solution (BSS), a non-functional mutant ANG (mANG), or wild-type ANG (ANG). The integrity of the blood-aqueous barrier was evaluated by the infiltrating cell and protein concentrations in aqueous humor. Histopathology, Western blot, and real-time qRT-PCR of aqueous humor and ocular tissue were performed to analyze inflammatory cytokines and transcription factors. EIU treated with ANG had decreased inflammatory cells and protein concentrations in the anterior chamber. Compared to BSS and mANG, ANG treatment showed reduced expression of IL-1ß, IL-8, TNF-α, and Myd88, while the expression of IL-4 and IL-10 was increased. Western blot of ANG treatment showed decreased expression of IL-6, inducible nitric oxide synthase (iNOS), IL-1ß, TNF-α, and phosphorylated NF-κB and increased expression of IL-10. In conclusion, ANG seems to reduce effectively immune mediated inflammation in the EIU rat model by reducing the expression of proinflammatory cytokines, while increasing the expression of anti-inflammatory cytokines through pathways related to NF-κB. Therefore, ANG shows potential for effectively suppressing immune-inflammatory responses in vivo.

Effects of botulinum toxin type A on the treatment of dry eye disease and tear cytokines.

PURPOSE: To determine the effects of botulinum toxin type A (BTX-A) injection on dry eye signs, symptoms, and tear cytokine levels in patients with intractable dry eye disease (DED). METHODS: In this prospective study, patients with intractable DED were randomized to a BTX-A (group A) or control group (group B). Patients were injected with BTX-A or normal saline in the medial part of the upper and lower eyelids. Before and at 2 weeks, 1 month, 2 months, and 4 months after injection, dry eye signs; tear film break-up time (TBUT), Schirmer I test, corneal fluorescein staining (CFS), and symptoms; ocular surface disease index (OSDI); and frequency of lubricants were assessed. The tear levels of matrix metalloproteinase (MMP)-9 and serotonin were measured before and at 1 month after injection. RESULTS: Fifty-two eyes from 26 patients (mean age, 57.7 years) were included. The TBUT was higher at 2 weeks and at 1 month in group A. The Schirmer I test and OSDI scores were also better in group A for up to 2 months. The CFS grades in group A were significantly lower until 4 months. Repeated measures analysis of variance (RMANOVA) demonstrated significant differences between the two groups over time for the Schirmer I test (p = 0.002), CFS (p = 0.025), OSDI (p = 0.020), and frequency of lubricants (p = 0.029). The MMP-9 conversion rate of group A (76.92%) was significantly higher than that of group B (38.46%, p = 0.005). The tear serotonin level in group A was reduced from 2.76 ± 0.34 to 1.73 ± 0.14 ng/mL (p < 0.001). No complications were observed during the study. CONCLUSION: BTX-A injection into the medial part of eyelid improves dry eye signs and symptoms and reduces tear cytokine levels. BTX-A is thus a potential treatment option for patients with intractable DED.

3D Architectures of Cox P Using Silk Fibroin Scaffolds: An Active and Stable Electrocatalyst for Hydrogen Generation in Acidic and Alkaline Media.

Developing nonprecious, highly active, and stable catalysts is essential for efficient electrocatalytic hydrogen evolution reaction in water splitting. In this study, the facile synthesis of a 3D flower-like Cox P/carbon architecture is proposed composed of an assembly of nanosheets interconnected by silk fibroin that acts as 3D scaffolds and a carbon source. This unique 3D architecture coupled with a carbon matrix enhances catalytic activity by exposing more active sites and increasing charge transport. The flower-like Cox P/carbon can facilitate a lower overpotential, Tafel slope, charge transfer resistance, and a higher electrochemically active surface than carbon-free and silk-free Cox P. The nanostructured architecture exhibits excellent catalytic performance with low overpotentials of 109 and 121 mV at 10 mA cm-2 and Tafel slopes of 55 and 62 mV dec-1 in acidic and alkaline media, respectively. Furthermore, it minimally degrades the overpotential and current density after long-term stability tests 10 000 cyclic voltammetry cycles and a chronoamperometric test over 40 h, respectively, in acidic media, which confirms the high durability and stability of the flower-like Cox P/carbon.

Ribonuclease 5 coordinates signals for the regulation of intraocular pressure and inhibits neural apoptosis as a novel multi-functional anti-glaucomatous strategy.

Glaucoma is a vision-threatening disorder characterized by progressive death of retinal ganglion cells (RGCs), although little is known about therapeutic milestones. Due to its complex and multifactorial pathogenesis, multipronged therapeutic approach is needed. Angiogenin (ANG), now called ribonuclease (RNase) 5, has been previously known as angiogenic factor and more recently its biologic activity is extended to promoting cell survival via its ribonucleolytic activity. Here, we revealed the defect of ANG in human glaucomatous trabecular meshwork (TM) cells and identified novel multiple functions of ANG as an anti-glaucomatous strategy. ANG was highly expressed in normal eyes and normal TM cells compared to glaucomatous TM cells. ANG induced intraocular pressure (IOP) lowering in rat models of both normal and elevated IOP, and as a possible mechanism, activated Akt-mediated signals for nitric oxide (NO) production, an important regulator of IOP in glaucomatous TM cell. Moreover, we demonstrated ANG-induced production of matrix metalloproteinase (MMP)-1 and -3 and rho-kinase inhibition for TM remodeling. For anti-glaucomatous defense optimization, ANG not only elicited immune-modulative pathways via indolamine 2,3-dioxygenase (IDO) activation in TM cells and suppression of Jurkat T cells, but also rescued neural stem cells (NSCs) from apoptosis induced by glaucomatous stress. These results demonstrate that novel multi-functional effects of ANG may have benefits against glaucoma in ocular tissues.

TNF-α upregulates HIF-1α expression in pterygium fibroblasts and enhances their susceptibility to VEGF independent of hypoxia.

The clinical manifestations of pterygium are characterized by rapid growth and postoperative recurrences. We had previously proposed that hypoxia-inducible factor (HIF)-1α recruits progenitor cells during the development and progression of pterygia. Recently, it was reported that various stimuli, including inflammation, could activate HIF-1α even under normoxic conditions. The ocular surface directly faces external environments, and is thus frequently exposed to inflammatory insults. First, we examined the gene expression of HIF-1α, its downstream molecule, vascular endothelial growth factor (VEGF)-A, and VEGF receptor (VEGFR)-2 in corneal and conjunctival cells compared with cultured human umbilical vein endothelial cells. Corneal fibroblasts had high expression of VEGFR-2 in the presence of TNF-α, and HIF-1α was activated by TNF-α in diverse ocular surface cells. The HIF-1α/VEGF/VEGFR signaling pathway in response to TNF-α was evaluated in cultured human pterygium fibroblasts (HPFs) at the gene and protein levels and was compared to treatment with cobalt chloride (CoCl2), a hypoxic mimetic, to exclude the effect of hypoxia. Although VEGF-A expression was not changed by TNF-α, expression of HIF-1α and VEGFR-2 was enhanced in HPFs treated with TNF-α, independent of hypoxia conditioning. In addition, VEGF-C gene expression was activated solely by TNF-α in HPF, but VEGF-B levels were not significantly affected. These results may provide mechanistic explanations for the uniquely vigorous proliferation of pterygium fibrovascular tissue during TNF-α-induced ocular surface inflammation.

Pulmonary glass particles may persist in the lung suppressing function of immune cells.

The health effects of silica may depend on the inherent properties of crystalline silica or on external factors affecting the biological activity or distribution of its polymorphs. Inhaled crystalline silica is classified as a Group I carcinogen, however, information on the health effects of amorphous silica is still insufficient. Considering that alveolar macrophages play a key role in both innate and adaptive immune responses for removal of foreign bodies that enter via the respiratory system, we treated sheet-like glass particles (SGPs), a type of noncrystalline amorphous silica, to MH-S cells, an alveolar macrophage cell line. SGPs reduced the generation of ROS and NO and induced cell death via multiple pathways. Although the expression of CD80, CD86, and CD40, increased by exposure to SGPs, the expression of MHC class II molecules had not notably changed. Additionally, expression of ICAM-1 tended to decrease. In mice, SGPs were distributed in the interstitial region of the lung without notable pathological lesion on day 14 after a single intratracheal instillation. Pulmonary total cell number increased significantly with the highest dose, but the levels of all measured inflammatory cytokines and chemokines, except IL-1, were lower in BAL fluid from SGP-treated mice compared to control. More interestingly, the expression of antigen presentation-related proteins was enhanced in the lungs of SGP-exposed mice concomitant with an increase in the number of mature dendritic cells, whereas the expression of ICAM-1, an important adhesion molecule for helper T cell recruitment, was suppressed. Taken together, we suggest that SGPs may induce adverse health effects by down-regulating function of immune cells in the lungs. Furthermore, ICAM-1 may play a key role in immune response to remove pulmonary SGPs.

Fibroblast biology in pterygia.

Activation of fibroblasts is a vital process during wound healing. However, if prolonged and exaggerated, profibrotic pathways lead to tissue fibrosis or scarring and further organ malfunction. Although the pathogenesis of pterygium is known to be multi-factorial, additional studies are needed to better understand the pathways initiated by fibroblast activation for the purpose of therapeutic translation. Regarding pterygium as a possible systemic disorder, we discuss the different cell types that pterygium fibroblasts originate from. These may include bone marrow-derived progenitor cells, cells undergoing epithelial-mesenchymal transition (EMT), and local resident stromal cells. We also describe how pterygium fibroblasts can be activated and perpetuate profibrotic signaling elicited by various proliferative drivers, immune-inflammation, and novel factors such as stromal cell-derived factor-1 (SDF-1) as well as a known key fibrotic factor, transforming growth factor-beta (TGF-ß). Finally, epigenetic modification is discussed to explain inherited susceptibility to pterygium.

Angiogenin ameliorates corneal opacity and neovascularization via regulating immune response in corneal fibroblasts.

BACKGROUND: Angiogenin (ANG), a component of tears, is involved in the innate immune system and is related with inflammatory disease. We investigated whether ANG has an immune modulatory function in human corneal fibroblasts (HCFs). METHODS: HCFs were cultured from excised corneal tissues. The gene or protein expression levels of interleukin (IL)-1beta (ß), IL-4, IL-6, IL-8, IL-10, complements, toll-like receptor (TLR)4, myeloid differentiation primary response gene (MYD)88, TANK-binding kinase (TBK)1, IkappaB kinase-epsilon (IKK-ε) and nuclear factor-kappaB (NF-κB) were analyzed with or without ANG treatment in tumor necrosis factor-alpha (TNF-α)- or lipopolysaccharide (LPS)-induced inflammatory HCFs by real-time polymerase chain reaction (PCR), Western blotting and immunocytochemistry. Inflammatory cytokine profiles with or without ANG were evaluated through immunodot blot analysis in inflammatory HCFs. Corneal neovascularization and opacity in a rat model of corneal alkali burn were evaluated after application of ANG eye drops. RESULTS: ANG decreased the mRNA levels of IL-1ß, IL-6, IL-8, TNF-α receptor (TNFR)1, 2, TLR4, MYD88, and complement components except for C1r and C1s and elevated the mRNA expression of IL-4 and IL-10. Increased signal intensity of IL-6, IL-8 and monocyte chemotactic protein (MCP)-1 and MCP-2 induced by TNF-α or LPS was weakened by ANG treatment. ANG reduced the protein levels of IKK-ε by either TNF-α and LPS, and decreased TBK1 production induced by TNF-α, but not induced by LPS. The expression of NF-κB in the nuclei was decreased after ANG treatment. ANG application lowered corneal neovascularization and opacity in rats compared to controls. CONCLUSION: These results demonstrate that ANG reduces the inflammatory response induced by TNF-α or LPS in HCFs through common suppression of IKK-ε-mediated activation of NF-κB. This may support the targeting of immune-mediated corneal inflammation by using ANG.

Biomineralized multifunctional magnetite/carbon microspheres for applications in Li-ion batteries and water treatment.

Advanced functional materials incorporating well-defined multiscale architectures are a key focus for multiple nanotechnological applications. However, strategies for developing such materials, including nanostructuring, nano-/microcombination, hybridization, and so on, are still being developed. Here, we report a facile, scalable biomineralization process in which Micrococcus lylae bacteria are used as soft templates to synthesize 3D hierarchically structured magnetite (Fe3O4) microspheres for use as Li-ion battery anode materials and in water treatment applications. Self-assembled Fe3O4 microspheres with flower-like morphologies are systematically fabricated from biomineralized 2D FeO(OH) nanoflakes at room temperature and are subsequently subjected to post-annealing at 400 °C. In particular, because of their mesoporous properties with a hollow interior and the improved electrical conductivity resulting from the carbonized bacterial templates, the Fe3 O4 microspheres obtained by calcining the FeO(OH) in Ar exhibit enhanced cycle stability and rate capability as Li-ion battery anodes, as well as superior adsorption of organic pollutants and toxic heavy metals.

Oncologic results and prognostic predictors of patients with locally advanced rectal cancer showing ypN0 after radical surgery following neoadjuvant chemoradiotherapy.

PURPOSE: Adjuvant chemotherapy is recommended for patients with locally advanced rectal cancer after radical surgery following neoadjuvant chemoradiotherapy (NCRT) regardless of the final pathologic stage. However, the efficacy of adjuvant chemotherapy in ypN0 patients remains controversial. The aim of this study was to evaluate the oncologic outcomes and analyze the prognostic factors for ypN0 patients in order to estimate prognosis and establish an effective adjuvant chemotherapy strategy for stage 0-II rectal cancers after radical surgery following NCRT. METHODS: Between January 1999 and December 2009, the medical records of 202 patients who had been diagnosed with locally advanced rectal cancer, underwent radical surgery following NCRT, and showed ypN0 were retrospectively reviewed. RESULTS: The median follow-up period was 60.5 months. The 5-year local recurrence rate was 3.1 %. The 5-year disease-free survival and 5-year overall survival were 86.3 and 86.9 %. Postirradiation T3-4 and abdominoperineal resection (APR) were the independent prognostic indicators for disease-free survival (p = 0.001, p = 0.003) and overall survival (p = 0.001, p = 0.002). Adjuvant chemotherapy improved local recurrence in the patient with ypT3-4 and patients who had undergone APR (p = 0.014, p = 0.002). APR affected local recurrence, disease-free survival, and overall survival of ypT3-4 patients (p = 0.013. 0.029, and 0.001) CONCLUSIONS: Postirradiation T3-4 and APR are the significant prognostic factors for ypN0. Further randomized prospective study is needed to evaluate the oncologic benefit of adjuvant chemotherapy in ypN0 patients, especially those with ypT3-4 and those having undergone APR, and to confirm which chemotherapeutic agent could improve the oncologic outcomes of patients poorly responding to NCRT.

Angiogenin reduces immune inflammation via inhibition of TANK-binding kinase 1 expression in human corneal fibroblast cells.

Angiogenin (ANG) is reportedly multifunctional, with roles in angiogenesis and autoimmune diseases. This protein is involved in the innate immune system and has been implicated in several inflammatory diseases. Although ANG may be involved in the anti-inflammatory response, there is no evidence that it has direct anti-inflammatory effects. In this study we sought to determine whether ANG has an anti-inflammatory effect in human corneal fibroblasts (HCFs) exposed to media containing tumor necrosis factor-alpha (TNF-α). We found that ANG reduced the mRNA expression of interleukin-1 beta (IL-1ß), -6, -8 and TNF-α receptors (TNFR) 1 and 2. In contrast, ANG increased the mRNA expression of IL-4 and -10. Protein levels of TANK-binding kinase 1 (TBK1) were reduced by ANG in HCFs treated with TNF-α. Moreover, ANG diminished the expression of IL-6 and -8 and monocyte chemotactic protein- (MCP-) 1. The protein expression of nuclear factor-κB (NF-κB) was downregulated by ANG treatment. These findings suggest that ANG suppressed the TNF-α-induced inflammatory response in HCFs through inhibition of TBK1-mediated NF-κB nuclear translocation. These novel results are likely to play a significant role in the selection of immune-mediated inflammatory therapeutic targets and may shed light on the pathogenesis of immune-mediated inflammatory diseases.

Evaluation of stem cell components in retrocorneal membranes.

The purpose of this study was to elucidate the origin and cellular composition of retrocorneal membranes (RCMs) associated with chemical burns using immunohistochemical staining for primitive cell markers. Six cases of RCMs were collected during penetrating keratoplasty. We examined RCMs with hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) staining and immunohistochemical analysis using monoclonal antibodies against hematopoietic stem cells (CD34, CD133, c-kit), mesenchymal stem cells (beta-1-integrin, TGF-ß, vimentin, hSTRO-1), fibroblasts (FGF-ß, α-smooth muscle actin), and corneal endothelial cells (type IV collagen, CD133, VEGF, VEGFR1). Histologic analysis of RCMs revealed an organized assembly of spindle-shaped cells, pigment-laden cells, and thin collagenous matrix structures. RCMs were positive for markers of mesenchymal stem cells including beta-1-integrin, TGF-ß, vimentin, and hSTRO-1. Fibroblast markers were also positive, including FGF-ß and α-smooth muscle actin (SMA). In contrast, immunohistochemical staining was negative for hematopoietic stem cell markers including CD34, CD133 and c-kit as well as corneal endothelial cell markers such as type IV collagen, CD133 except VEGF and VEGFR1. Pigment-laden cells did not stain with any antibodies. The results of this study suggest that RCMs consist of a thin collagen matrix and fibroblast-like cells and may be a possible neogenetic structure produced from a lineage of bone marrow-derived mesenchymal stem cells.

Influence of modified Furlow double opposing Z-plasty on mandibular growth in Oriental patients with cleft palate and/or lip.

The purpose of this study was to statistically analyze mandibular growth disturbance in the Oriental cleft population and compare this with that of non-cleft children through cephalometric analysis. Thirty-six children with cleft palates, repaired using a modification of Furlow double opposing Z-plasty, were chosen. Comparative analyses among 3 types of cleft were performed. Statistical analyses of 8 linear and angular measurements were performed in cleft patients and the non-cleft population using Fisher Z-transformation. Comparative analysis showed no significant difference among the 3 types of cleft. In the Oriental cleft group treated with modified double opposing Z-plasty, the spatial position of the mandible showed significant differences compared with the non-cleft group. A backward inclination of the anterior surface of the mandible and downward rotation of the mandibular body were identified. Some of our results regarding gonial angle and length of the mandibular body conflicted with previous Western studies.

The results and complications of cranial bone reconstruction in patients with craniosynostosis.

The aim of this study was to report the treatment results and complications of cranial bone reconstruction in patients with craniosynostosis at the Seoul National University Children's Hospital over the 14-year period from 1996 to 2009. A retrospective study was undertaken on 96 cases of 94 consecutive patients diagnosed with craniosynostosis, operated on between 1996 and 2009. The authors collected the data regarding age at surgery, operative time, duration of hospitalization, and follow-up period. Surgical results including the amount of blood loss, signs related to increased intracranial pressure, aesthetic results, cranial index, and complications were evaluated. The authors compared these results with those of the past 10-year period from 1986 to 1995 at the same hospital. There were 81 patients with single synostosis and 13 patients with multiple synostoses. The age of patients ranged from 3 months to 130 months (mean, 27.9 months). The mean operative time was 7.54 ± 1.77 hours. The duration of hospital stay ranged from 5 to 70 days with a mean period of 8.97 days. The intraoperative loss of hemoglobin ranged from 0 to 9.2 g/dL (mean, 2.35 g/dL). The symptoms of increased intracranial pressure were improved after cranioplasty. With regard to aesthetic results, 94 patients were classified into category I and 2 into category IV. The cranial index decreased from 88.0% ± 1.2% to 86.9% ± 1.1% on average. There were 34 minor complications in 28 operations (29.2%). There were statistically significant differences in age at surgery, operative time, duration of hospital stay, and reoperation rate between the recent 14-year study and the past 10-year study. We observed good surgical results and low complications of cranial bone reconstruction in patients with craniosynostosis. The results of the recent 14-year operations were better than those of the past 10-year operations.

Human mesenchymal stem cells differentiate into keratocyte-like cells in keratocyte-conditioned medium.

Culturing corneal keratocytes is difficult because keratocytes growing in a monolayer rapidly lose their stellate morphology and cease to express keratocyte markers such as keratocan, lumican and aldehyde dehydrogenase 1 family, member A1 (ALDH1A1). Conversely, mesenchymal stem cells (MSCs) can be easily expanded in cell culture, and they have a variety of differentiation pathways. We studied the feasibility of using MSCs as a source for corneal tissue engineering. Based on the observation that keratocytes have MSC-like properties, similar to bone marrow-derived MSCs (BM-MSCs), we hypothesized that MSCs would differentiate into corneal keratocyte-like cells in keratocyte-conditioned medium (KCM). We measured changes in the expression of keratocyte markers through quantitative real-time polymerase chain reaction (qRT-PCR) and found that human MSC's cultured in KCM expressed both keratocan and ALDH1A1. Western blot analysis demonstrated that human MSCs cultured in KCM steadily increased their expression of lumican and ALDH1A1, while they lost expression of α-smooth muscle actin (α-SMA). Immunocytochemistry indicated that human MSCs grown in KCM acquired characteristics similar to those of keratocytes. These results suggest that KCM can direct human MSCs to differentiate into keratocyte-like cells.

Superior long-term cycling stability of SnO2 nanoparticle/multiwalled carbon nanotube heterostructured electrodes for Li-ion rechargeable batteries.

We demonstrate the fabrication of hybrid nanocomposite electrodes with a combination of SnO(2) nanoparticles (NPs) and conducting multiwalled carbon nanotube (MWCNT) anodes (SnO(2)@CNT) through the direct anchoring of SnO(2) NPs on the surface of electrophoretically pre-deposited MWCNT (EPD-CNT) networks via a metal-organic chemical vapor deposition process. This SnO(2)@CNT nanocomposite displays large reversible capacities of over 780, 510, and 470 mA h g(-1) at 1 C after 100, 500, and 1000 cycles, respectively. This outstanding long-term cycling stability is a result of the uniform distribution of SnO(2) NPs (~8.5 nm), a nanoscale EPD-CNT network with good electrical conductivity, and the creation of open spaces that buffer a large volume change during the Li-alloying/dealloying reaction of SnO(2).

Ocular surface discomfort and Demodex: effect of tea tree oil eyelid scrub in Demodex blepharitis.

The purpose of this study was to evaluate the relation between ocular discomfort and ocular Demodex infestation, and therapeutic effects of tea tree oil (TTO) in Demodex blepharitis patients. Three hundred and thirty-five patients with ocular discomfort were evaluated for ocular Demodex infestation and subjective symptoms with ocular surface discomfort index (OSDI) score. Among them, Demodex-infested patients were randomized to receive either eyelid scrubbing with TTO (TTO group,106 patients) or without TTO (Control group, 54 patients) for 1 month. Demodex were found in 84% of patients with ocular discomfort. The number of Demodex was significantly correlated with age (P = 0.04) and OSDI score (P = 0.024). After eyelid scrub treatment, Demodex count was reduced from 4.0 ± 2.5 to 3.2 ± 2.3 in the TTO group (P = 0.004) and from 4.3 ± 2.7 to 4.2 ± 2.5 in the control group (P = 0.27). Also, OSDI score was reduced from 34.5 ± 10.7 to 24.1 ± 11.9 in the TTO group (P = 0.001) and from 35.3 ± 11.6 to 27.5 ± 12.8 in the control group (P = 0.04). In conclusion, Demodex number showed a significant positive correlation with age and subjective ocular discomfort. The tea tree oil eyelid scrub treatment is effective for eliminating ocular Demodex and improving subjective ocular symptoms.

Tear cytokines and chemokines in patients with Demodex blepharitis.

PURPOSE: The purpose of the study is to evaluate the causes of inflammation in Demodex-induced blepharitis by analyzing cytokine levels in lacrimal fluid. METHODS: Fifteen Demodex blepharitis patients were selected for assessment of tear cytokine concentrations. Fifteen Demodex-free blepharitis patients and 15 subjects with no ocular symptoms were selected as control groups. Minimally stimulated tear samples (20µl) were collected from each eye and analyzed using a Luminex® 200™ Total System for detection of IL-1ß, IL-5, IL-7, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor (G-CSF), and macrophage inflammatory protein-1 beta (MIP-1ß). RESULTS: The concentration of IL-17 in tears was significantly higher in the Demodex blepharitis group than in the Demodex-free blepharitis group. Tear IL-7 and IL-12 levels show serial increases for these three groups (p<0.05). There were no significant differences in the other cytokines levels between both blepharitis groups. We confirmed that elevated cytokines normalized after treatments. CONCLUSIONS: Infestation of Demodex mites induces change of tear cytokine levels, IL-17 especially, which cause inflammation of the lid margin and ocular surface. These findings might increase our understanding of the mechanism of ocular discomfort and telangiectasias frequently found in Demodex blepharitis patients.

Clinical and immunological responses in ocular demodecosis.

The purpose of this study was to investigate clinical and immunological responses to Demodex on the ocular surface. Thirteen eyes in 10 patients with Demodex blepharitis and chronic ocular surface disorders were included in this study and treated by lid scrubbing with tea tree oil for the eradication of Demodex. We evaluated ocular surface manifestations and Demodex counts, and analyzed IL-1ß, IL-5, IL-7, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor, and macrophage inflammatory protein-1ß in tear samples before and after the treatment. All patients exhibited ocular surface manifestations including corneal nodular opacity, peripheral corneal vascularization, refractory corneal erosion and infiltration, or chronic conjunctival inflammatory signs before treatment. After treatment, Demodex was nearly eradicated, tear concentrations of IL-1ß and IL-17 were significantly reduced and substantial clinical improvement was observed in all patients. In conclusion, we believe that Demodex plays an aggravating role in inflammatory ocular surface disorders.

Caveolin-1 as a novel indicator of wound-healing capacity in aged human corneal epithelium.

Excess caveolin-1 has been reported to play a role in age-dependent hyporesponsiveness to growth factors in vitro. Therefore, we hypothesized that caveolin-1-dependent hyporesponsiveness to growth factors in aged corneal epithelial cells might be responsible for delayed wound healing in vivo. To test this hypothesis, we evaluated corneal wound-healing time by vital staining using fluorescein after laser epithelial keratomileusis (LASEK). We compared wound-healing times in young, middle-aged and elderly patients. We also examined caveolin-1 levels and other aging markers, such as p53 and p21, in the corneal epithelium. Elderly patients generally had higher caveolin-1 levels in the corneal epithelia than young patients. There were, however, variations among individuals with increased caveolin-1 in some young patients and decreased levels in some elderly patients. Wound-healing time after LASEK correlated well with the corneal caveolin-1 status. Therefore, we suggest that caveolin-1 status might be responsible for delayed wound healing in elderly patients after LASEK. Caveolin-1 status might be a regulator for wound-healing capacity and a novel target for in vivo adjustment.