Mass Spectrometric Analyses of Phosphatidylcholines in Alkali-Exposed Corneal Tissue

Crane, Ashley M.; Hua, Hong-Uyen; Coggin, Andrew; Gugiu, Bogdan G.; Lam, Byron L.; Bhattacharya, Sanjoy K.

doi:10.1167/iovs.12-10448

Cited by 22 publications

(22 citation statements)

References 29 publications

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“…Sourcing of TM samples from distant locations contributed to significant unavoidable transit times and storage in PBS or Optisol GS. Our control analyses with cornea 16 and other anterior chamber tissue (data not shown) from mammalian model systems (porcine, bovine, and a select subset of humans) showed across the board decrease of phospholipid species, but no selective absence or appearance of a phospholipid species as a function of storage up to a week at 48C in PBS or Optisol GS (Bhattacharya SK, Aribindi K, Crane AC, unpublished observations, 2012). Three other factors are intrinsically likely to affect the lipid profiles broadly apart from biochemical individuality of the donors: (1) confounding factors of diseases or disorders that are well known to affect lipid profiles, such as hyperlipidemia; (2) use of drugs by the donors, such as statins, and (3) confounding factor of diseases that are not lipid-related per se, such as diabetes.…”

Section: Discussionmentioning

confidence: 79%

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Comparative Phospholipid Profiles of Control and Glaucomatous Human Trabecular Meshwork

Aribindi¹,

Guerra²,

Lee³

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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Section: Discussionmentioning

confidence: 79%

“…The procedures to determine extraction efficiency are similar to that which were performed for our published corneal lipid extraction and analyses. 16 …”

Section: Lipid Extractionmentioning

confidence: 99%

Comparative Phospholipid Profiles of Control and Glaucomatous Human Trabecular Meshwork

Aribindi¹,

Guerra²,

Lee³

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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“…AQH samples were subjected to only one thawing cycle prior to their use. Our control analyses with cornea and other anterior chamber tissue/fluids from mammalian model systems (porcine, bovine and a select subset of humans) showed a decrease of phospholipid species but no selective absence or appearance of a phospholipid species as a function of storage up to a week at 4°C in PBS or Optisol GS [49] and no alteration when immediately stored in −80°C and thawed only once (Bhattacharya, SK and Aribindi, K.; unpublished observations), our observations parallels that for mass spectrometric analyses of serum [50], where repeated freeze-thaw cycles have been previously shown to affect small molecules and peptides. AQH is likely to provide an accurate depiction into the differential lipid profile between control and diseased samples because of the local control of sample from collection to analyses.…”

Section: Discussionmentioning

confidence: 99%

Phospholipid profiles of control and glaucomatous human aqueous humor

et al. 2014

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To compare phospholipid (phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine and phosphatidylinositol) profiles of human control and glaucomatous aqueous humor (AQH). AQH samples were procured during surgery from human POAG and control subjects (n=15 each). Samples were used following institutional review board approved protocols and adhering to the tenets of the Declaration of Helsinki. Lipid extraction was performed using a modification of the Bligh and Dyer method, protein concentrations were determined using the Bradford’s method, and select samples were confirmed with Densitometry of PHAST gels. Lipids were identified and subjected to ratiometric quantification using a TSQ Quantum Access Max triple quadrupole mass spectrometer utilizing precursor ion scan (PIS) or neutral ion loss scan (NLS) using appropriate class specific lipid standards in a two step quantification process. The comparative profiles of phosphatidylcholines, phosphatidylserines, phosphatidylethanolamines and phosphatidylinositols between control and glaucomatous AQH showed several species common between them. A number of unique lipids in all four phospholipid classes were also identified in control eyes that were absent in glaucomatous eyes and vice versa. A number of phospholipids were found to be uniquely present in control, but absent in glaucomatous AQH and vice versa. Compared with a previous study of control and POAG red blood cells, a number of these phospholipids are absent locally (AQH).

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“…Proteins recovered from the corresponding upper aqueous phase were quantified using Bradford’s method [27]. In order to ensure extraction efficiency, an external standard (10 picomoles of 1, 2-ditridecanoyl-sn-glycero-3-phosphocholine) was premixed with AH prior to lipid extraction [13, 28]. All extractions and subsequent handling were done using glass vials to avoid contaminating impurities.…”

Section: Methodsmentioning

confidence: 99%

Sphingolipids and ceramides of mouse aqueous humor: Comparative profiles from normotensive and hypertensive DBA/2J mice

et al. 2014

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Purpose To identify the sphingolipid and ceramide species and their quantitative differences between normotensive and hypertensive intraocular pressure states in DBA/2J mouse aqueous humor (AH). Methods Normotensive and hypertensive AH was sampled from mice by paracentesis. Lipid extraction was performed using modifications of the Bligh and Dyer method. Protein concentration was estimated using the Bradford colorimetric assay. Sphingolipids and ceramides were identified and subjected to ratiometric quantification using appropriate class specific lipid standards on a TSQ Quantum Access Max triple quadrupole mass spectrometer. Results The comparative profiles of normotensive and hypertensive DBA/2J mouse AH showed several species of sphingomyelin, sphingoid base, sphingoid base-1-phosphate (S1P) and ceramides common between them. A number of unique lipids in each of the above lipid classes were also identified in normotensive AH that were absent in hypertensive AH and vice versa. Conclusion A number of sphingolipid and ceramide species were found to be uniquely present in normotensive, but absent in hypertensive AH and vice versa. Further pursuit of these findings is likely to contribute towards expanding our understanding of the molecular changes associated with increased intraocular pressure (IOP) and glaucoma.

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Mass Spectrometric Analyses of Phosphatidylcholines in Alkali-Exposed Corneal Tissue

Abstract: Alkali exposure dramatically changes the PC profile of cornea. Our data are consistent with penetration and hydrolysis as stochastic contributors to changes in PCs due to exposure to alkali for a finite duration and amount.

Cited by 22 publications

References 29 publications

Comparative Phospholipid Profiles of Control and Glaucomatous Human Trabecular Meshwork

Comparative Phospholipid Profiles of Control and Glaucomatous Human Trabecular Meshwork

Phospholipid profiles of control and glaucomatous human aqueous humor

Sphingolipids and ceramides of mouse aqueous humor: Comparative profiles from normotensive and hypertensive DBA/2J mice

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