Lysophospholipids and lysophospholipase D in rabbit aqueous humor following corneal injury.

We previously found that lysophosphatidic acid (LPA)-like activity eliciting Cl(-) currents in Xenopus oocytes is increased in rabbit aqueous humor (AH) following corneal freeze wounds. The purpose of this study was to examine whether actual levels of LPA in AH from wounded eyes are higher than those from control eyes, and to determine the sources and enzymatic pathways of AH LPA in control and wounded conditions. Lysophospholipase D (lysoPLD) activity was measured by the enzymatic determination of choline following incubation of AH samples with exogenous lysophosphatidylcholines (LPCs). The molecular species compositions of LPA and LPC in fresh and incubated AH were determined by liquid chromatography-tandem mass spectrometry. A high, but similar activity of lysoPLD in the samples from both control and freeze-wounded eyes was detected. Its enzymatic properties resemble those of plasma lysoPLD, identified as autotaxin. Levels of LPCs, predominant substrates of lysoPLD in AH, were several times higher in the AH samples from injured eyes than those from the control eyes. Our results suggest that lysoPLD is constitutively released from corneal tissues and/or ciliary body into the AH, with no injury-induced increase in release following freeze-wounding. They also suggest that wound-induced increases in LPA-like biological activity are due to linoleoyl species-rich molecular composition in AH from wounded eyes. A possible mechanism of the altered molecular composition is an increase in the AH concentrations of LPCs, linoleoyl species of which are preferentially converted to corresponding unsaturated LPA by the constitutively active lysoPLD.

Quantification of subarachnoid hemorrhage by three-dimensional computed tomography: correlation between hematoma volume and symptomatic vasospasm.

Subarachnoid hemorrhage (SAH) volume was measured by three-dimensional computed tomography (3D-CT) and the correlation examined between the SAH volume and the occurrence of symptomatic vasospasm (SVS). Experimental (in vitro) hematomas were made with blood obtained from 10 volunteers. The hematoma volume was determined by actual measurements and by 3D-CT using a CT number in the range of 40-80 Hounsfield units (HU) on days 1, 4, 7, 11, and 14. The coefficients on days 1 and 4 were relatively high and the correlation between measured and estimated volumes was significant on days 7, 11, and 14. 3D-CT was also performed in 50 patients with SAH at onset (day 0) and on days 1, 4, 7, and 14. The hematoma volume including the volume of normal structures was automatically calculated (V1). The volume of normal structures (V2) with CT numbers of 40-80 HU was calculated in another 50 patients without intracranial lesions as 12 ml. The total hematoma volume was defined as V1 minus mean V2. The mean SAH volume was 44, 36, 21, 11, and 8 ml on days 0, 1, 4, 7, and 14, respectively. The hematoma volumes were significantly larger in patients with SVS than in patients without SVS at all time points. The minimum hematoma volume in patients with SVS was 92, 76, 42, 24, and 12 ml on days 0, 1, 4, 7, and 14, respectively. This method allows the quantitative determination of SAH volume based on 3D-CT, and may be useful in clinical studies of cerebral vasospasm.