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The abundance of clusterin in pterygium is illustrated by immunofluorecence. Labeling for clusterin is in red, nuclei are blue (DAPI). The arrow indicates cells enveloping the body of the pterygium that are clusterin negative.
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Pterygium is a vision-impairing fibrovascular lesion that grows across the corneal surface and is associated with sunlight exposure. To increase our understanding of the cells types involved in pterygium, we have used expressed sequence tag analysis to examine the transcriptional repertoire of isolated pterygium and to identify marker genes for tis...
Citations
... 55 The protein levels of S100A6 were previously observed up-regulated in dry eye disease and Meibomian gland dysfunction 32,56 and down-regulated in the tear film of patients diagnosed with keratoconus 57 and were therefore proposed as candidate biomarkers of these ocular surface disorders. Interestingly, S100A6 was found to be up-regulated in experimental corneal neovascularization models 58 and expressed in pterygium tissue removed from patient, 59,60 but no studies related this protein with AMD disease to date. ...
Purpose:
Specific altered metal homeostasis has been investigated in the tear film of age-related macular degeneration (AMD) patients considering that metal dyshomeostasis contributes to the production of free radicals, inflammation, and apoptosis and results in conformational changes of proteins.
Methods:
A multitargeted approach based on spectrophotometry and mass spectrometry techniques has been implemented to the multiplexed quantitation of lactoferrin (LF), S100 calcium binding protein A6 (S100A6), metallothionein 1A (MT1A), complement factor H (CFH), clusterin (CLU), amyloid precursor protein (APP), Mg, P, Na, Fe, Cu, Zn, and Ca, in the tear film from 60 subjects, 31 patients diagnosed with the dry form of AMD, and 29 healthy individuals.
Results:
Significant up-regulations of MT1A (1.9-fold) and S100A6 (1.4-fold) and down-regulations of LF (0.7-fold), Fe (0.6-fold), Mg (0.7-fold), and Cu (0.7-fold) were observed in AMD patients, when compared to control subjects. Of all the studied variables, only APP showed negative correlation with age in the AMD group. Also, positive correlations were observed for the variables Mg and Na, Cu and Mg, and P and Mg in both the AMD and control groups, whereas positive correlations were exclusively determined in the AMD group for Cu and LF, Na and Ca, and Mg and Ca. The panel constituted of MT1A, Na, and Mg predicts AMD disease in 73% of cases.
Conclusions:
The different levels of target metals and (metallo-)proteins in the tear film suggest altered metal homeostasis in AMD patients. These observed pathophysiological changes may be related with the anomalous protein aggregation in the macula.
... Gene expression profiling is an unbiased approach which can provide clues for hypothesis development. Several studies have used gene expression microarray platforms or profiling of expressed sequence tags to compare differentially expressed genes (DEGs) in pterygium and healthy conjunctiva [17][18][19][20][21][22][23]. Very recently, RNA sequencing (RNA-seq) technology was applied for the first time [24,25]. ...
... We then researched the top DEG function by accessing information in GeneCards Suite (Weizmann Institute of Life Science, Rehovot, Israel), and by literature searches on PubMed (National Institutes of Health (NIH), National Library of Medicine (NLM), National Center for Biotechnology Information (NCBI)). As an assessment of reproducibility, we compared our findings to those of the previously published gene expression profiling studies noted in the Introduction to this paper [18][19][20]22,24,25], as well as to studies that investigated expression of individual genes (discussed more below). ...
... Thus, of the top 25 upregulated DEGs in pterygium-E, 21 were also upregulated in pterygium-NE (84% similarity). Of the top 25 upregulated DEGs in pterygium-NE, 22 were also upregulated in pterygium-E (88% similarity). ...
Solar damage due to ultraviolet radiation (UVR) is implicated in the development of two proliferative lesions of the ocular surface: pterygium and pinguecula. Pterygium and pinguecula specimens were collected, along with adjacent healthy conjunctiva specimens. RNA was extracted and sequenced. Pairwise comparisons were made of differentially expressed genes (DEGs). Computational methods were used for analysis. Transcripts from 18,630 genes were identified. Comparison of two subgroups of pterygium specimens uncovered evidence of genomic instability associated with inflammation and the immune response; these changes were also observed in pinguecula, but to a lesser extent. Among the top DEGs were four genes encoding tumor suppressors that were downregulated in pterygium: C10orf90, RARRES1, DMBT1 and SCGB3A1; C10orf90 and RARRES1 were also downregulated in pinguecula. Ingenuity Pathway Analysis overwhelmingly linked DEGs to cancer for both lesions; however, both lesions are clearly still benign, as evidenced by the expression of other genes indicating their well-differentiated and non-invasive character. Pathways for epithelial cell proliferation were identified that distinguish the two lesions, as well as genes encoding specific pathway components. Upregulated DEGs common to both lesions, including KRT9 and TRPV3, provide a further insight into pathophysiology. Our findings suggest that pterygium and pinguecula, while benign lesions, are both on the pathological pathway towards neoplastic transformation.
... e former acts as a defense against UV-induced oxidative stress on the ocular surface [13], while PRDX2 contributes as part of the antioxidant defense by inactivating hydrogen peroxide [14]. Specific forms of keratin, including keratin 13 and 4, are increased in pterygium and have a role in cell migration [15]. ese three biomarkers were detected in the primary isolated HPF of this work. ...
Background:
Sedum dendroideum has antioxidant effects that are beneficial for different diseases. We aimed to analyze the antiproliferative activity of S. dendroideum in human pterygium fibroblasts (HPFs).
Methods:
HPFs were treated for 24 h with 0-1000 μg/mL of S. dendroideum lyophilized to analyze its effect on cell viability using the CellTiter assay. RNA from HPF treated with 250 μg/mL of S. dendroideum lyophilized was isolated, and the expression of VEGF and CTGF genes was evaluated by qPCR. A dermal fibroblast cell line (HDFa) was used as a healthy control. The total phenolic content, antioxidant activity, and chemical profile of S. dendroideum lyophilized were determined.
Results:
Viability of HPF decreased after 24 h treatment of S. dendroideum in a dose-dependent manner. The expression of VEGF and CTGF significantly decreased (P < 0.01) in HPF treated with 250 μg/mL of S. dendroideum when compared with untreated HPF. The total phenolic concentration in the S. dendroideum lyophilized was 33.67 mg gallic acid equivalents (GAE)/g. Antioxidant activity was 384.49 mM Trolox equivalents/mL. The main phenolic compounds identified by HPLC analysis were the kaempferol-3-O-glycoside, kaempferol-3-O-rhamnoside, kaempferol-3-O-neohesperidoside-7-O-α-rhamnopyranoside, and kaempferol-3-O-glycoside-7-O-rhamnoside.
Conclusions:
S. dendroideum decreases the proliferation of HPF and the expression of VEGF and CTGF. The phenolic compound concentration, antioxidant activity, and phytochemical profile may play a role in these effects.
... The top four expressed keratins, KRT12, KRT13, KRT14, KRT15, showed differential expression patterns among the epithelial clusters (Fig. 6g). KRT13 was enriched in the bulbar conjunctival epithelium and KRT12 was enriched in the corneal epithelium, a pattern consistent with previous reports 31,32,66,67 . Meesmann dystrophy presents with unstable and fragile corneal epithelium, and is associated with KRT3 or KRT12 mutations, both of which are corneal epithelial differentiation markers and not highly-expressed in the basal or limbal epithelium 37,68 . ...
Bulk RNA sequencing of a tissue captures the gene expression profile from all cell types combined. Single-cell RNA sequencing identifies discrete cell-signatures based on transcriptomic identities. Six adult human corneas were processed for single-cell RNAseq and 16 cell clusters were bioinformatically identified. Based on their transcriptomic signatures and RNAscope results using representative cluster marker genes on human cornea cross-sections, these clusters were confirmed to be stromal keratocytes, endothelium, several subtypes of corneal epithelium, conjunctival epithelium, and supportive cells in the limbal stem cell niche. The complexity of the epithelial cell layer was captured by eight distinct corneal clusters and three conjunctival clusters. These were further characterized by enriched biological pathways and molecular characteristics which revealed novel groupings related to development, function, and location within the epithelial layer. Moreover, epithelial subtypes were found to reflect their initial generation in the limbal region, differentiation, and migration through to mature epithelial cells. The single-cell map of the human cornea deepens the knowledge of the cellular subsets of the cornea on a whole genome transcriptional level. This information can be applied to better understand normal corneal biology, serve as a reference to understand corneal disease pathology, and provide potential insights into therapeutic approaches.
... Rekürrens oranının değerlendirilmesi sonuç açısından öncelikli ölçüm değeriydi. Rekürrens, limbus sınırını aşan yeni fibrovasküler büyüme olarak tanımlandı (8). ...
... Rekürrens oranının değerlendirilmesi sonuç açısından öncelikli ölçüm değeriydi. Rekürrens, limbus sınırını aşan yeni fibrovasküler büyüme olarak tanımlandı (8). ...
INTRODUCTION: To evaluate the results of limbal conjunctival autograft transplantation (LCAT) in patients with primary and recurrent pterygium. METHODS: Forty-two eyes of 38 patients were included in the study. 26 of them were primary, while 16 were recurrent pterygium. Limbal conjunctival autograft transplantation was performed after the pterygium excision to all eyes. Patients having a follow-up of at least 12 months were evaluated with respect to demographic data, pterygium grade, surgical complications, recurrence rate, refractive astigmatism and visual acuity. RESULTS: The mean age of the patients was 44 ± 8.5. Twenty-six (68.4%) patients were male and 12 (31.6%) were female. No recurrence were observed in primary pterygium group, whereas two recurrences were observed in recurrent pterygium group. The overall recurrence rate was 4.7%. Buttonhole complication occurred while forming the graft in three patients. Conjunctival inclusion cyst developed in 2 patients with recurrent pterygium. DISCUSSION AND CONCLUSION: Limbal conjunctival autograft transplantation seems to be an effective and safe surgical option in both primary and recurrent pterygiums.
GİRİŞ ve AMAÇ: Primer ve rekürren pterjiyumlu olgularda gerçekleştirdiğimiz limbal konjonktival otogreft transplantasyonu (LCAT) sonuçlarını değerlendirmek. YÖNTEM ve GEREÇLER: Çalışmaya 38 hastanın 42 gözü dahil edildi. Bunlardan 26'sı primer iken 16'sı rekürren pterjiyumdu. Tüm gözlere pterjiyum eksizyonu ile birlikte limbal konjonktival otogreft transplantasyonu uygulandı. En az 12 ay boyunca takip edilen hastalar, demografik veriler, pterjiyum evresi, cerrahi komplikasyonlar, nüks oranı, refraktif astigmatizma ve görme keskinliği açısından değerlendirildi. BULGULAR: Hastaların ortalama yaşı 44 ± 8.5 idi. Hastaların 26'sı (% 68.4) hasta erkek, 12'si (% 31.6) kadındı. Primer pterjium grubunda nüks görülmezken, tekrarlayan pterjium grubunda iki nüks gözlendi. Toplam nüks oranı %4.7 idi. Üç hastada greft oluşturulurken düğme iliği (buttonhole) komplikasyonu meydana geldi. Rekürren pterjiyumlu 2 hastada konjonktival inklüzyon kisti gelişti. TARTIŞMA ve SONUÇ: Hem primer hem de rekürren pterjiyum tedavisinde limbal konjonktival otogreft transplantasyonu etkili ve güvenilir bir cerrahi seçenek olarak gözükmektedir.
... UV exposure of pterygium epithelial cells generates proinflammatory cytokines, such as IL6 and IL8 (Di Girolamo et al., 2006). In addition, UV-B-driven oxidative stress can induce other inflammatory factors, like IL1α and TNFα that can lead other delirious signaling by activating MMPs Also, the progression of a pterygium may be the result of excessive wound healing, cell proliferation, and a limbal stem-cell deficiency triggered by oxidative stress due to UV exposure, and/or an inflammatory mediator, immunologic mechanisms, growth factors, or viruses on the ocular surface (Coroneo et al., 1999;Di Girolamo et al., 2004;Jaworski et al., 2009;Threlfall and English, 1999). The parts of pterygium are head (apical part present in cornea), neck (limbal part) and body (scleral part). ...
A sight threatening, pterygium is a common ocular surface disorders identified by fibrovascular growth of the cornea and induced by variety of stress factors, like ultraviolet (UV) exposure. However, the genes involved in the etiopathogenesis of this disease is not well studied. Herein, we identified the gene expression pattern of pterygium and examined the expression of pterygium-related genes in UV-B-induced human primary cultured corneal epithelial cells (HCEpCs), telomerase immortalized human corneal epithelial (hTCEpi), primary conjunctival fibroblast (HConFs) and primary pterygium fibroblast cells (HPFCs). A careful analysis revealed that the expression of 10 genes was significantly modulated (by > 10-fold). Keratin 24 (KRT24) and matrix metalloproteinase 9 (MMP-9) were dramatically upregulated by 49.446- and 24.214-fold, respectively. Intriguingly, UV-B exposure (50 J/m²) induced the upregulation of the expressions of MMP-9 in corneal epithelial cells such as HCEpCs and hTCEpi. Furthermore, UV-B exposure (100 and/or 200 J/m²) induced the upregulation of the expressions of MMP-9 in fibroblast such as HConFs and HPFCs. The exposure of HCEpCs to 100 and 200 J/m² UV-B induced significant expressions of KRT24 mRNA. Nevertheless, no expression of KRT24 mRNA was detected in HConFs and HPFCs. The findings provide evidence that the progression of pterygium may involve the modulation of extracellular matrix-related genes and vasculature development and the up-regulation of KRT24 and MMP-9 by UV stress. UV radiation may promote the modulation of these pterygium-related genes and induce the initiation and progression of human pterygium.
... Both K13 and K4 seems to be found as a protein in the exosomes in human urine [30], but nothing is known about the role of K4 as a signalling molecule. Curiously K4 is mentioned in a human study regarding an eye pathology, the pterygium, an overgrowth of fibrovascular tissue from conjuntiva across the corneal surface associated with sunlight exposure [31]. Interestingly, the most abundant molecules expressed in the study of Jaworski and colleagues [31], besides K4, are K13 and solute channels involved in mineral transport such as Aquaporin 3 (AQP3) as well as the S100A family, which are also implicated in our research. ...
... Curiously K4 is mentioned in a human study regarding an eye pathology, the pterygium, an overgrowth of fibrovascular tissue from conjuntiva across the corneal surface associated with sunlight exposure [31]. Interestingly, the most abundant molecules expressed in the study of Jaworski and colleagues [31], besides K4, are K13 and solute channels involved in mineral transport such as Aquaporin 3 (AQP3) as well as the S100A family, which are also implicated in our research. Recently K4, AQPs and the S100 cluster were inserted in the group of seasonal rhythm genes (SRGs) also in cashmere goat skin [21]. ...
Background:
Cashmere goat is known for its precious undercoat. Being photoperiod-dictated, cashmere growth has been studied focusing mainly on hair follicle cycle phases (anagen, catagen and telogen). An accurate molecular knowledge of the goat hair follicle cycle, disentangling gene expression changes during phases and recognizing timing boundaries, could be useful to improve cashmere goat management and ultimately cashmere production.
Results:
To better describe goat's hair follicle transcriptome we applied RNA-sequencing to isolated hair follicles from five Italian cashmere goats, during the anagen and catagen phase, identifying total of 214 differentially expressed genes (DEGs): 97 were up-regulated while 117 were down-regulated in catagen with respect to anagen. Gene Ontology and pathway analysis were performed. We detected 144 significant pathways spanning from estrogen, pluripotency of stem cells, thermogenesis and fatty acid metabolism that were strongly expressed during the hair follicle phases analysed. Finally, we validated promising DEGs by RT-qPCR in the same set of samples as well as in hair follicles and entire skin biopsies of another cashmere goats cohort accounting for early anagen, anagen, early catagen, and catagen phases.
Conclusions:
As in the isolated hair follicles, some target genes were homogenously modulated during the four hair follicle phases. Ceruloplasmin (CP) and Keratin 4 (K4), confirmed their clear cut expression between growing and resting phase. In fact, K4 was almost absent in catagen phases while CP was barely expressed in anagen phases. In particular, the strong expression of K4 in early anagen makes it an eligible marker to track the beginning of a new hair cycle, and therefore defining the optimum time for cashmere harvesting.
... The human CYP1A1 gene is located on chromosome 15q22-24, composed of seven exons responsible for procarcinogen activation of reactive metabolites and is involved in oxidation mediation, including polycyclic aromatic hydrocarbons and aromatic amino to mutagenic and carcinogenic metabolites. CYP1A1 genetic polymorphism may cause changes or loss of CYP1A1 enzyme activity, and result in cellular detoxification damage, which can lead to the absence of cell growth suppression mechanisms [21,22]. ...
CYP1A1 gene, a carcinogenic metabolisms enzymes encoded gene, was previously found to be detected in pterygium tissue. We aimed to determine the association between CYP1A1 m1 (rs4646903) polymorphisms with CYP1A1 enzymes, p53 protein, and vascular endothelial growth factor (VEGF) level in patients with inflammatory and non-inflammatory pterygium. DNA isolation was performed from a blood sample of 70 pterygium patients consisting of 35 inflammatory and 35 non-inflammatory pterygia. Rs 4646903 SNP Genotyping T> C (m1) in the CYP1A1 gene was performed using restriction fragment length polymorphisms-PCR (RFLP-PCR). PCR products confirmed and sent to Macrogen, South Korea for sequencing. Polymorphism results are characterized as wild type (TT), mutant homozygote (TC), and mutant heterozygote (CC). CYP1A1 gene polymorphisms consist of mutant heterozygote (TC), mutant homozygote (CC) and wild type (TT). In both groups, the heterozygote mutant was higher than the wild type and mutant homozygote. The CYP1A1 enzyme level was higher in inflammatory pterygium, P53 protein levels were higher in the non-inflammatory group, and VEGF levels were higher in the inflammatory group. CYP1A1 polymorphisms were not associated with CYP1A1 enzyme levels, p53 protein levels, and VEGF in both groups. CYP1A1 gene polymorphism has not been shown to be associated with levels of CYP1A1 enzymes, p53 and VEGF in both pterygium groups.
... It is important to note, the exogenous ECM that was injected sub-conjunctivally in the experiment contained the heparin sulfate proteoglycan. Histopathological evaluation of pterygium demonstrates a variety of findings, such as squamous metaplasia of epithelial cells and goblet cell hyperplasia and fibrovascular connective tissue overgrowth with elastotic degenerative alterations of the connective tissue, while prominent neovascularization and inflammatory cell infiltration was also observed [14,15]. Significant changes both in the epithelium and in the underlying connective tissue and diffuse immune cell infiltrate has been shown by Golu et al. [16]. ...
Pterygium is a triangle-shaped fibrovascular hyperplasia of the bulbar conjunctiva on the cornea. The purpose of this study was to analyze Proteoglycans (PGs) by Immunohistochemistry (IHC) in pterygium tissues and to compare the results with normal conjunctiva. Twenty-four patients (14 males) undergoing primary pterygium excision and 17 healthy individuals (10 males), undergoing extracapsular cataract surgery, were included. Pterygium tissues and normal conjunctiva tissues were surgically removed. The tissue sections were fixed in 2% paraformaldehyde and incubated with monoclonal antibodies against PGs anti-mouse IgG. Immunohistochemical study showed stronger expression of keratan sulfate in the stroma of the pterygium compared to normal conjunctiva. An increased expression of heparan sulfate was observed in the epithelial layer and around the pterygium vessels. On the other hand, dermatan sulfate showed an increased expression and localization not only in the sub-epithelial area of the pterygium and normal conjunctiva, yet throughout the stroma of the pterygium. The differences in the expression and localization of the studied extracellular matrix proteoglycans in the pterygium tissue compared to normal conjunctiva may explain the tissue hyperplasia, structure, and the functional properties in pterygium.
