OBJECTIVE To determine the effects of grape seed extract (GSE), lutein, and fish oil containing omega-3 fatty acids on oxidative stress, migration, proliferation, and viability of lens epithelial cells (LECs). SAMPLE Lens capsules or cultured LECs obtained from canine cadavers. PROCEDURES An antioxidant reductive capacity assay was used to determine reducing capability of each substance. The LECs were cultured and incubated with various substances, including N-acetyl cysteine (NAC), when appropriate, and dimethyl sulfoxide (DMSO) as positive and vehicle control substances, respectively. A dichlorofluorescein assay was used to evaluate reactive oxygen species (ROS) production, and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to determine cell viability. Ex vivo posterior capsule opacification (PCO) was used to evaluate LEC migration and proliferation. RESULTS Antioxidant reductive effects of GSE surpassed those of NAC, lutein, and fish oil containing omega-3 fatty acids. The GSE reduced ROS production in LECs, compared with the DMSO vehicle control, whereas lutein was pro-oxidative. All test substances reduced cell viability. Ex vivo PCO was not altered by GSE, was decreased by lutein, and was increased by fish oil containing omega-3 fatty acids, compared with results for the DMSO vehicle control. CONCLUSIONS AND CLINICAL RELEVANCE Only GSE had significant antioxidant capabilities and reduced ROS production; however, no effect on ex vivo PCO was detected. Fish oil containing omega-3 fatty acids increased ex vivo PCO. No conclusions could be made regarding antioxidant effects of these substances on LECs. These findings suggested that the substances will not decrease PCO.

It has been established that L-gamma-glutamylated derivatives of alpha-amino acids are delivered more efficiently to the kidneys than are the parent alpha-amino acids. Therefore, we synthesized L-gamma-glutamyl-S-(1,2-dichlorovinyl)-L-cysteine (L-gamma-glutamyl-L-DCVC), the simplest L-gamma-glutamylated derivative of the nephrotoxic alpha-amino acid S-(1,2-dichlorovinyl)-L-cysteine (L-DCVC), and investigated its effects on renal function and ultrastructure in pentobarbital-anesthetized dogs. Intravenous doses of 23.15 and 92.60 micromoles of L-gamma-glutamyl-L-DCVC/kg of body weight induced significant increases in urinary protein output and significant decreases in the clearance of inulin during the 6-hour post-injection period. Changes were not observed in any of the other 13 renal function variables or in the 11 plasma and blood variables that were monitored throughout the same period. Both doses of L-gamma-glutamyl-L-DCVC induced renal ultrastructural lesions in the S1 and S2 cells of the canine proximal tubule; the remaining 8 cell types downstream and the glomeruli were not damaged. The onset and magnitude of renal function changes and the cell types affected by L-gamma-glutamyl-L-DCVC were virtually identical to those observed previously following IV administration of equivalent doses of L-DCVC to pentobarbital-anesthetized dogs. Rapid removal of the L-gamma-glutamyl group from L-gamma-glutamyl-L-DCVC (ie, deglutamylation) resulting in formation of the parent alpha-amino acid, L-DCVC, can best explain the extreme similarity in the nephrotoxic profiles of these 2 toxicants.

OBJECTIVE To characterize aminoaciduria and plasma amino acid concentrations in dogs with hepatocutaneous syndrome (HCS). ANIMALS 20 client-owned dogs of various breeds and ages. PROCEDURES HCS was definitively diagnosed on the basis of liver biopsy specimens (n = 12), gross and histologic appearance of skin lesions (4), and examination of skin and liver biopsy specimens (2) and presumptively diagnosed on the basis of cutaneous lesions with compatible clinicopathologic and hepatic ultrasonographic (honeycomb or Swiss cheese pattern) findings (2). Amino acid concentrations in heparinized plasma and urine (samples obtained within 8 hours of each other) were measured by use of ion exchange chromatography. Urine creatinine concentration was used to normalize urine amino acid concentrations. Plasma amino acid values were compared relative to mean reference values; urine-corrected amino acid values were compared relative to maximal reference values. RESULTS All dogs had generalized hypoaminoacidemia, with numerous amino acid concentrations < 50% of mean reference values. The most consistent and severe abnormalities involved glutamine, proline, cysteine, and hydroxyproline, and all dogs had marked lysinuria. Urine amino acids exceeding maximum reference values (value > 1.0) included lysine, 1-methylhistidine, and proline. CONCLUSIONS AND CLINICAL RELEVANCE Hypoaminoacidemia in dogs with HCS prominently involved amino acids associated with the urea cycle and synthesis of glutathione and collagen. Marked lysinuria and prolinuria implicated dysfunction of specific amino acid transporters and wasting of amino acids essential for collagen synthesis. These findings may provide a means for tailoring nutritional support and for facilitating HCS diagnosis.

This paper describes the development of ∨99mTc tricarbonyl cysteine as potential renal function diagnostic radiopharmaceutical and evaluation of its biological characteristics using experimental animals. ℓ-Cysteine was labeled efficiently with ∨99mTc tricarbonyl precursor ([∨99mTc(CO)₃(H₂O)₃)]+) under 30 min heating at 75℃. Labeling yield and stability were analyzed by high performance liquid chromatography (HPLC). The biodistribution property of ∨99mTc tricarbonyl cysteine in mice and its dynamic imaging profiles in rabbits were carried out.

The proteolytic activities present in midguts of both fed and unfed Haemaphysalis longicornis were assessed by using the gelatin-substrate gel electrophoresis and inhibitor sensitivity analyses. Three predominant (116, 48 and 40 kDa) and two weak (55 and 60 kDa) proteinase bands were commonly expressed in both unfed and fed ticks, while a weak 80 kDa band was only present in fed ticks. Consistent with observations on other tick species, proteolytic activity against the gelatin substrate was observed only under acidic conditions. Inhibition studies against the gelatin substrate using a panel of inhibitors showed that the predominant proteolytic enzymes of 40 and 48 kDa molecular mass are cysteine proteinases. These results are discussed in the context of host vaccination as an alternative tick control method to the current use of chemical acaricides