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ABSTRACT: The Ca2+-independent phospholipase A2 (iPLA2) subfamily of enzymes is associated with arachidonic acid (AA) release and the subsequent increase in fatty acid turnover.
This phenomenon occurs not only during apoptosis but also during inflammation and lymphocyte proliferation. In this study,
we purified and characterized a novel type of iPLA2 from bovine brain. iPLA2 was purified 4,174-fold from the bovine brain by a sequential process involving DEAE-cellulose anion exchange, phenyl-5PW
hydrophobic interaction, heparin-Sepharose affinity, Sephacryl S-300 gel filtration, Mono S cation exchange, Mono Q anion
exchange, and Superose 12 gel filtration. A single peak of iPLA2 activity was eluted at an apparent molecular mass of 155 kDa during the final Superose 12 gel-filtration step. The purified
enzyme had an isoelectric point of 5.3 on twodimensional gel electrophoresis (2-DE) and was inhibited by arachidonyl trifluoromethyl
ketone (AACOCF3), Triton X-100, iron, and Ca2+. However, it was not inhibited by bromoenol lactone (BEL), an inhibitor of iPLA2, and adenosine triphosphate (ATP). The spot with the iPLA2 activity did not match with any known protein sequence, as determined by matrix-assisted laser desorption/ionization time-of-flight
(MALDI-TOF) analysis. Altogether, these data suggest that the purified enzyme is a novel form of cytosolic iPLA2.
Keywordsbrain–Ca2+-independent PLA2
–characterization–purification
Molecules and Cells 04/2012; 32(5):405-413. · 2.18 Impact Factor
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ABSTRACT: The Ca(2+)-independent phospholipase A(2) (iPLA(2)) subfamily of enzymes is associated with arachidonic acid (AA) release and the subsequent increase in fatty acid turnover. This phenomenon occurs not only during apoptosis but also during inflammation and lymphocyte proliferation. In this study, we purified and characterized a novel type of iPLA(2) from bovine brain. iPLA(2) was purified 4,174-fold from the bovine brain by a sequential process involving DEAE-cellulose anion exchange, phenyl-5PW hydrophobic interaction, heparin-Sepharose affinity, Sephacryl S-300 gel filtration, Mono S cation exchange, Mono Q anion exchange, and Superose 12 gel filtration. A single peak of iPLA(2) activity was eluted at an apparent molecular mass of 155 kDa during the final Superose 12 gel-filtration step. The purified enzyme had an isoelectric point of 5.3 on two-dimensional gel electrophoresis (2-DE) and was inhibited by arachidonyl trifluoromethyl ketone (AACOCF(3)), Triton X-100, iron, and Ca(2+). However, it was not inhibited by bromoenol lactone (BEL), an inhibitor of iPLA(2), and adenosine triphosphate (ATP). The spot with the iPLA(2) activity did not match with any known protein sequence, as determined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) analysis. Altogether, these data suggest that the purified enzyme is a novel form of cytosolic iPLA(2).
Molecules and Cells 08/2011; 32(5):405-13. · 2.18 Impact Factor
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ABSTRACT: We evaluated the long-term outcome of simultaneous transurethral bladder tumor and prostate resection in patients with nonmuscle invasive bladder tumor and bladder outlet obstruction.
Between April 1997 and April 2006, 213 patients with nonmuscle invasive bladder tumor who had a minimum followup of 24 months were included in the study, including group 1-107 with transurethral resection of bladder tumor only and group 2-106 with transurethral bladder tumor and prostate resection. Simultaneous transurethral bladder tumor and prostate resection was performed at surgeon discretion. The records were retrospectively analyzed for clinicopathological parameters, recurrence and progression rates, time to recurrence and postoperative uroflowmetry results in the 2 groups.
There were no significant differences in clinicopathological parameters between the 2 groups. At a mean followup of 54.3 and 50.1 months in groups 1 and 2, respectively, group 2 patients with a tumor less than 3 cm or a single tumor had a significantly lower recurrence rate than group 1 patients. None of the 31 patients with recurrence in group 2 had recurrence in the bladder neck or prostatic urethra where transurethral prostate resection had been done. There was no significant difference in the progression rate between the 2 groups. The 60-month recurrence-free probability in groups 1 and 2 was 43.4% and 52.0%, respectively. Three months after surgery the postvoid residual urine volume had significantly decreased in group 2.
Simultaneous transurethral bladder tumor and prostate resection may help decrease bladder cancer recurrence and delay time to recurrence without the risk of cancer implantation when transurethral prostate resection is done, especially in patients with a papillary, solitary-appearing bladder lesion less than 3 cm.
The Journal of urology 03/2009; 181(4):1594-9; discussion 1599. · 4.02 Impact Factor
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ABSTRACT: We evaluated the collagen-to-smooth muscle tissue matrix ratio and percentage of elastin in the renal pelvis, ureteropelvic junction (UPJ) and ureter, and compared these findings with the degree of obstruction, patient age and post-pyeloplasty renal recovery.
We analyzed histological sections from 75 patients with UPJ obstruction. Nine patients were excluded owing to bilateral UPJ obstruction and an improper specimen. We divided the specimen obtained from pyeloplasty into 3 parts, namely the renal pelvis above the obstruction, the obstructed UPJ portion and the ureter below the obstruction. To examine the collagen and smooth muscle, sections were stained using Masson's trichrome, and elastic van Giesson stain was used for elastin, smooth muscle and collagen. Collagen, smooth muscle and elastin populations were identified, and the tissue matrix ratio and percentage of elastin were calculated by color image analysis.
In patients with lower ratios of collagen-to-smooth muscle in the UPJ proper hydronephrosis was more improved postoperatively (p = 0.049). In patients with a lower percentage of elastin in the renal pelvis, UPJ and ureter hydronephrosis was more improved postoperatively (p <0.0001).
Because the UPJ portion was resected during pyeloplasty, the renal pelvis and the ureter remaining after pyeloplasty are likely to be related to improved hydronephrosis. A higher percentage of elastin in the renal pelvis and ureter contributes to inelasticity and low compliance, and results in a slower recovery from hydronephrosis after pyeloplasty.
The Journal of Urology 04/2005; 173(3):962-6. · 3.75 Impact Factor
