Quantification of tear proteins by SDS-PAGE with an internal standard protein: A new method with special reference to small volume tears

Li, Kaijun; Chen, Ziyan; Duan, Fang; Liu, Jingwen; Wu, Kaili

doi:10.1007/s00417-009-1275-3

Cited by 33 publications

(31 citation statements)

References 31 publications

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“…In contrast, galectin-3 was present in tears from the patients with ocular inflammation, which may have caused release of galectin-3 for self-healing. For example, the level of galectin-3 in tears from dry eye patients is 0.38 ng/μg total protein (Uchino et al, 2015); or 0.76–4.56 μg galectin-3/mL tear assuming 2–12 μg total protein/μL (Ng et al 2000; Li et al, 2010). Since partially degraded galectin-3 has been observed in 50% of patients (Uchino et al, 2015), the in vivo level of intact galectin-3 might be lower than the calculated and effective levels used in our models.…”

Section: Discussionmentioning

confidence: 99%

Galectin-3 enhances extracellular matrix associations and wound healing in monkey corneal epithelium

Fujii

Shearer

Azuma

2015

Experimental Eye Research

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Poor healing of epithelial wounds in cornea is a major clinical problem, leading to persistent epithelial defects and ulceration. The primary cause is poor cell migration over the wound. Carbohydrate-binding protein galectin-3 binds to extracellular matrixes (ECMs) and promotes lamellipodia formation by cross-linking to α3 integrin. Recombinant galectin-3 also facilitates wound healing in the rodent cornea. The purposes of the present experiments were to: (1) establish epithelial wound healing models in monkey corneal explant culture, the models more relevant to human, (2) evaluate the healing effect of galectin-3 in our models, and (3) determine if galectin-3 enhances cell adhesion by interacting with ECMs on corneal surface and their ligand integrins. Monkey corneas with central wounds produced by sodium hydroxide (NaOH) or n-heptanol were incubated with or without recombinant galectin-3. The defected area was stained with sodium fluorescein. Primary isolated corneal epithelial cells from monkey were cultured with or without galectin-3 on plates coated with ECMs or integrins, and the number of adhering cells was counted. Galectin-3 expression in various eye tissues was visualized by immunoblotting. NaOH caused loss of epithelial cells and basement membrane. n-Heptanol removed epithelial cells, but the basement membrane was retained. These corneal defects spontaneously became smaller in a time-dependent manner. Exogenous galectin-3 enhanced wound healing in both NaOH and n-heptanol models. Galectin-3 also enhanced cell adhesion onto the major ECMs found in the basement and Bowman’s membranes and onto integrins. Relatively high levels of galectin-3 were detected in corneal and conjunctival epithelium, but tear fluid contained negligible galactin-3. These results suggested that the enhanced binding of epithelial cells to ECMs and integrins caused by galectin-3 might promote cell migration over wounded corneal surfaces. Since tear fluid contained relatively low levels of galectin-3, exogenous galectin-3 may be a beneficial drug to enhance re-epithelialization in human corneal diseases.

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Section: Discussionmentioning

confidence: 99%

Galectin-3 enhances extracellular matrix associations and wound healing in monkey corneal epithelium

Fujii

Shearer

Azuma

2015

Experimental Eye Research

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“…Quantitative determination of tear proteins is of increasing interest in ophthalmology, but still there remains a technical problem due to small sample volumes available and the complexity of sample composition. 32 Tear sampling is definitely a major challenge and has the greatest significant influence on the precision and reproducibility of the analytical results.…”

Section: Tear Collection Methods and Their Application In Practicementioning

confidence: 99%

Evaluation of commonly used tear sampling methods and their relevance in subsequent biochemical analysis

et al. 2017

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The human precorneal tear film is a special body fluid, since it is a complex mixture of proteins, lipids, small bioactive molecules, and their concentrations and relative distribution represent not only the metabolic state of the ocular surface but also the systemic and local homeostasis of the outer eye and the human body. This suggests that biochemical analysis of the precorneal tear film composition may provide a non-invasive tool for diagnosis and monitoring of disease progression or treatment efficacy in human medicine. However, collecting tears is demanding, and obtaining reproducible and unaltered samples is challenging because of the small sample volumes of tears. Several methods are available for tear collection as a preparatory step of precorneal tear film analysis, and the collection method used has to be assessed since it has a critical impact on the effectiveness of the assays and on the quality of the results. Each sampling method has advantages and disadvantages; therefore, it is not easy to choose the appropriate collecting method for tear collection. To overcome these limitations various methods have been recommended by different authors for special aspects of specific tests. The aim of our review was to evaluate tear sampling methods with regard to our ongoing biochemical analysis. *Contributed equally.

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“…Proteomic analysis has become an important factor to biomedical research, since it is a valuable means of studying the healthy and diseased eye (Markoulli et al 2001, and correlates with systemic or ocular disease (De Souza et al 2006, Li et al 2010. The main focus in clinical proteomics is the discovery of new proteins or peptides that work as biomarkers correlated to a specific disease (Couture et al 2006, Grus et al 2007, Campos et al 2008.…”

Section: Introductionmentioning

confidence: 99%

“…Metalloproteinases were identified in the dog (Couture et al 2006) and equine tears from corneal injury (Ollivier et al 2004), and dogs with cancer showed a different expression pattern of proteins (Campos et al 2008). Quantitative determination of tear proteins is of increasing interest in ophthalmology, but remains a technical challenge due to the small sample volumes available and the complexity of its composition (Li et al 2010).…”

Section: Introductionmentioning

confidence: 99%

“…Different protein expression reported may be a result of true biological feature or from methodological variation (Lu et al 2010). Lowry (Li et al 2010) and Bradford (Sitaramamma et al 1998, Ananthi et al 2008, Li et al 2010) methods were used to quantify proteins from tear samples (Ananthi et al 2008, Lu et al 2010. For proteomic analysis, tear samples can be obtained from microcapillary tubes (Gachon et al 1982, Jones et al 1997, Couture et al 2006, Grus et al 2007, Green-Church et al 2008 or from Schirmer tear test strips (Grus et al 2007, Li et al 2008, Roberts & Erickson 2008.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of two methods of tear sampling for protein quantification by Bradford method

et al. 2013

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esquerdo, respectivamente. Para as amostras obtidas com STT os resultados foram 4,45mg/mL ±0,35 and 4,52mg/ mL ±0,29 para olhos direito e esquerdo, respectivamente. Diferenças estatisticamente significativas (p<0,001) foram encontradas entre os dois métodos. Nas condições em que o trabalho foi conduzido, a média da concentração proteica pelo método de Bradford das amostras obtidas através das tiras do STT foi superior à obtida com o tubo microcapilar. Os valores padrão da concentração protéica da lágrima obtida pelo método de Bradford devem ser analisados considerando-se o método de coleta da lágrima, uma vez que este interfere significativamente nos resultados obtidos.

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Quantification of tear proteins by SDS-PAGE with an internal standard protein: A new method with special reference to small volume tears

Cited by 33 publications

References 31 publications

Galectin-3 enhances extracellular matrix associations and wound healing in monkey corneal epithelium

Galectin-3 enhances extracellular matrix associations and wound healing in monkey corneal epithelium

Evaluation of commonly used tear sampling methods and their relevance in subsequent biochemical analysis

Comparison of two methods of tear sampling for protein quantification by Bradford method

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