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Javier Benitez-del-Castillo, CARMEN GALBIS-ESTRADA, Sheila Pons-Vazquez, TERESA REGI-PUERTAS, CARLA MARCO-RAMIREZ, Jose J. Garcia-Medina, MARIA D. PINAZO-DURAN; Simmultaneous Detection Of Multiple Cytokines In Women Tears. Invest. Ophthalmol. Vis. Sci. 2012;53(14):4243.
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Inflammatory cytokines play a role in a wide spectrum of ocular diseases. Molecular analyses of human tears have been difficult to assess by the small sample volume. We deal with simultaneously detecting multiple cytokines and chemokines in women tears to compare groups and subgroups regarding age, in an ophthalmologic population.
20-60 microliter tears were obtained from 27 healthy women attending the ophthalmologic departments with the aim of updating her refractive status. Tears were collected by a microcapillary tube with some gentle mechanical stimulation from the inferior fornix of both eyes, that were frozen (-80ºC) and stored in cryotubes until processing, by using multiplex cytometric bead-based optimized assay by means of bead a Luminex R100™. Statistical analyses was done by the SPSS 15.0 program.
Mean age was 46+13 years (ranged 26 to 80 years). Tears showed useful values in pg/mL for GM-CSF (8,33+3,13), IFN-G (2,17+1,27) , IL10 (4,93+3,71), IL12 (50,94+60,58) , IL1β (12,15+17,21) , IL2 (4,45+3,51) , IL4 (26,95+10,86) , IL5 (2,42+2,20), IL6 (13,34+9,83) , IL8 (532,75+601,04) , TNF-α (21,99+16,52), and VEGF (570,46+264,79). Significant differences in tear expression of all these molecules were observed in an age related fashion.
Differential expression of cytokine levels support tears as a useful indicator of molecular mechanisms occurring in the eyes. We suggest that the described methods can be suitable for large-scale screening of tear molecules for various ocular and systemic diseases.
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