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All Journal Indonesian Journal of Urology
Basuki B Purnomo, Basuki B
Lab. Ilmu Bedah/SMF Urologi, FK Universitas Brawijaya/RS Dr. Saiful Anwar, Malang
Health Professions Medicine & Pharmacology Public Health

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PERANAN ISOZIM PROTEIN KINASE C-ΑLFA DALAM MEMEDIASI EFEK ANTIPROLIFERASI ALFA-TOKOFEROL PADA SEL STROMA PROSTAT Purnomo, Basuki B
Indonesian Journal of Urology Vol 16 No 2 (2009)
Publisher : Indonesian Urological Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32421/juri.v16i2.334

Abstract

Objective: Alpha tocopherol, other than its anti-oxidant activity, also has a non anti-oxidant activity that correlates with inhibition of PKC-alpha isozyme activity. Several studies had confirmed its role in inhibition of vascular smooth muscle proliferation. Inhibition of stromal cell proliferation is an alternative method of prevention in BPH development. The aim of this in vitro study is to confirm that alpha-tocopherol, by its ability to inhibit PKC-alpha activity, has the capability to inhibit human prostatic stromal cell proliferation rate. Materials & Method: Primary cultures of human prostatic stromal muscle cells were obtained from 4 patients with bladder outlet obstruction caused by benign prostatic enlargement. The effects of various concentrations of alpha-tocopherol (20, 30, 40, 50, and 60 microM) were examined by cell proliferation and PKC-alpha isozyme activity assay. Cell proliferation study was performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. Caspase-3 activity assay was also performed to examine apoptotic mechanism. Results: MTT method, which is used to count viable cells, showed that exposures to various doses of alpha-tocopherol resulted in decreased proliferation rate. The decrement of proliferation rate is linear with the increment of alpha-tocopherol dose. Immunocytochemistry staining with anti caspase-3 antibody showed that no cells developed apoptosis. Alpha-tocopherol also inhibits PKC-alpha activity. Isozyme activity is reduced along with the increment of the dose (dose-dependent). There is a correlation between PKC-alpha activity with cell proliferation rate after exposure to alpha-tocopherol (R2=0,9279; p=0,000). Conclusion: Alpha-tocopherol at different concentrations reduces prostatic stromal cell proliferation rate, without effect on apoptosis. This antiproliferative property correlates with decreased activity of PKC-alpha isozyme.
INTRATESTICULAR INJECTION OF 20% HYPERTONIC SODIUM CHLORIDE AS A NOVEL CASTRATION METHOD: A PRECLINICAL STUDY Hendry, Marta; Hardjowijoto, Sunaryo; Purnomo, Basuki B; Joewarini, Endang; Meles, Dewa Ketut; P, Widodo J
Indonesian Journal of Urology Vol 17 No 1 (2010)
Publisher : Indonesian Urological Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32421/juri.v17i1.339

Abstract

Objectives: To demonstrate that intratesticular injection of 20% hypertonic sodium chloride can result in permanent castration and to evaluate serum changes in sodium chloride levels. Materials and Methods: A total of 40 Wistar rats were divided into 4 groups, consisting of bilateral orchidectomy (n=10), control (n=10) and 2 groups receiving intratesticular injections of 20% sodium chloride (n=10 in each group). Serum testosterone was measured on day 0, day 1, day 15 and day 30. Serum sodium chloride was assessed before injection, at 1 hour and 24 hours after injection. All testicles were harvested for histological examination. One way ANOVA and student t-test were used for statistical analysis. Results: Serum testosterone decreased to castrate levels in the orchidectomy and injected groups with no significant difference (p>0,05). Significant rise in serum sodium chloride was found 1 hour post injection (p<0,05) but after 1 day it decreased significantly (p<0,05). There was no significant difference in histopathological findings between the 2 injected groups after day 15 and 30 (p>0,05). Conclusion: Twenty percent hypertonic chloride injection has the same permanent castration effect with bilateral orchidectomy in rats. The serum sodium chloride changes did not reach the lethal level for rats. Therefore this treatment has a promising potential as a novel and cost-effective castration method with the additional benefit of retaining both testes.
PRODUKSI ANTIBODI POLIKLONAL HASIL INDUKSI ISOLAT PROTEIN NON KINASE DARI MEMBRAN SPERMATOZOA MANUSIA Lestari, Umie; Purnomo, Basuki B
Indonesian Journal of Urology Vol 15 No 2 (2008)
Publisher : Indonesian Urological Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32421/juri.v15i2.350

Abstract

Objective: The aim of this research is to get the profile of human sperms membrane protein based on identification for molecular weight, concentration protein, phosphorylation activity, and ability to raise enough antibodies. Material and methods: There are three phases in this research, first, isolation protein of human sperms membrane with using lysis buffer containing Tween-20, second, determination of molecular weight, phosphorylation activity, and protein concentration, and third, antibody production. Molecular weight of protein membrane was detected by SDS-PAGE, confirmation of protein concentration measured Biuret method, kinase activity measured using spectrophometry at its optimum condition, and then production of antibody that corfirmed by ELISA technique. Results: Human sperms contain the protein with molecular weight of 201, 163, 116, 97, 72, 56, 48, 32, 27, 20, and 17kDa. The highest phosphorylation activity was found on protein with molecular weight of 20 kDa is 30.197x 10-3 unit, while the lowest phosphorylation activity is 28.976x10-3 unit was shown on protein with 116 kDa of molecular weight and then   this protein can induce production of antibody. Biuret method show the protein concentration from protein with molecular weight 116 kDa is 410 ug/mL. Conclusion: The protein that has molecular weight of 116 kDa can be used as immunocontraception agent because phosphorylation activity has lower, good concentration, and could induce the production of antibody.
EFEK PAPARAN GLUKOSA KONSENTRASI TINGGI TERHADAP PENINGKATKAN AKTIVITAS ISOZIM PROTEIN KINASE C (PKC)-ALPHA PADA KULTUR SEL STROMA PROSTAT Purnomo, Basuki B; Widharse, I Wayan; Aulani’am, Aulani’am
Indonesian Journal of Urology Vol 16 No 1 (2009)
Publisher : Indonesian Urological Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32421/juri.v16i1.355

Abstract

OBJECTIVE: To prove that glucose treatment in human prostatic stromal cells (HPSC) culture would increase PKC alpha isozyme activity. MaterialS & Methods: Primary cultures of HPSC were obtained from patients with bladder outlet obstruction caused by benign prostatic enlargement. The effects of various concentrations of glucose (5, 10, 15, 20, 25, 30, 35 micro-Mol) on PKC-alpha isozyme activity were examined quantitatively by spectrophotometry methods. ResultS: Spectrophotometry analysis, which is used to assess isozyme activity quantitatively, showed that exposures to various doses of glucose resulted in increased isozyme activity. The increment of activity is linear with the increment of glucose dose. There is a strong correlation between glucose doses and PKC-alpha activity, with R2 = 0,91. Conclusion: At different concentrations glucose increases PKC-alpha isozyme activity of HPSC.
Co-Authors Aulani’am, Aulani’am Endang Joewarini I Wayan Widharse, I Wayan Marta Hendry, Marta Meles, Dewa Ketut Sunaryo Hardjowijoto Umie Lestari Widodo J P