Testosterone regulates myosin II isoforms expression and functional activity in the rat prostate.

BACKGROUND: Benign prostatic hyperplasia (BPH) is mainly caused by increased prostatic smooth muscle (SM) tone and prostatic volume. At the molecular level, SM myosin II (SMM II) and non-muscle myosin II (NMM II) mediate SM tone and cell proliferation while testosterone (T) plays a permissive role in the development of BPH. AIMS: The novel objective of this study was to elucidate the effects of T on the proliferation and apoptosis of rat prostatic cells and SM contractility as well as related regulatory signaling pathways. MATERIALS AND METHODS: Briefly, 36 male rats were divided into three groups (sham-operated, surgically castrated, and castrated with T supplementation). In vitro organ bath studies, competitive RT-PCR, Western-blotting analysis, Masson's trichrome staining, and immunofluorescence staining were performed. RESULTS: Our data showed that castration dramatically increased prostatic SM contractility and SM MHC immunostaining revealed a relatively increased SM cell numbers in the stroma. T deprivation altered prostate SMM II isoform composition with upregulation of SM-B and SM2 but downregulation of LC17a, favoring a faster more phasic-type contraction. Moreover, protein expressions of MLCK, p-MLCP, RhoB, ROCK1, and ROCK2 increased in castrated rats. Meanwhile NMM II heavy chain isoforms A, B, and C (NMMHC-A, B, and C isoforms) were altered by castration which may be linked to decreased cell proliferation and increased apoptosis. CONCLUSION: Our novel data demonstrated T regulates SMM II and NMM II and their functional activities in rat prostate and T ablation not only decreases prostate size (static component) but also changes the prostatic SM tone (dynamic component).

Blebbistatin modulates prostatic cell growth and contrapctility through myosin II signaling.

To investigate the effect of blebbistatin (BLEB, a selective myosin inhibitor) on regulating contractility and growth of prostate cells and to provide insight into possible mechanisms associated with these actions. BLEB was incubated with cell lines of BPH-1 and WPMY-1, and intraprostatically injected into rats. Cell growth was determined by flow cytometry, and in vitro organ bath studies were performed to explore muscle contractility. Smooth muscle (SM) myosin isoform (SM1/2, SM-A/B, and LC17a/b) expression was determined via competitive reverse transcriptase PCR. SM myosin heavy chain (MHC), non-muscle (NM) MHC isoforms (NMMHC-A and NMMHC-B), and proteins related to cell apoptosis were further analyzed via Western blotting. Masson's trichrome staining was applied to tissue sections. BLEB could dose-dependently trigger apoptosis and retard the growth of BPH-1 and WPMY-1. Consistent with in vitro effect, administration of BLEB to the prostate could decrease rat prostatic epithelial and SM cells via increased apoptosis. Western blotting confirmed the effects of BLEB on inducing apoptosis through a mechanism involving MLC20 dephosphorylation with down-regulation of Bcl-2 and up-regulation of BAX and cleaved caspase 3. Meanwhile, NMMHC-A and NMMHC-B, the downstream proteins of MLC20, were found significantly attenuated in BPH-1 and WPMY-1 cells, as well as rat prostate tissues. Additionally, BLEB decreased SM cell number and SM MHC expression, along with attenuated phenylephrine-induced contraction and altered prostate SMM isoform composition with up-regulation of SM-B and down-regulation of LC17a, favoring a faster contraction. Our novel data demonstrate BLEB regulated myosin expression and functional activity. The mechanism involved MLC20 dephosphorylation and altered SMM isoform composition.

Degradation of 2,6-di-tert-butylphenol by an isolated high-efficiency bacterium strain.

An aerobic bacterium strain, F-3-4, capable of effectively degrading 2, 6-ditert-butylphenol (2, 6-DTBP), was isolated and screened out from an acrylic fiber wastewater and the biofilm in the wastewater treatment facilities. This strain was identified as Alcaligenes sp. through morphological, physiological and biochemical examinations. After cultivation, the strain was enhanced by 26.3% in its degradation capacity for 2,6-DTBP. Results indicated that the strain was able to utilize 2,6-DTBP, lysine, lactamine, citrate, n-utenedioic acid and malic acid as the sole carbon and energy source, alkalinize acetamide, asparagine, L-histidine, acetate, citrate and propionate, but failed to utilize glucose, D-fructose, D-seminose, D-xylose, serine and phenylalanine as the sole carbon and energy source. The optimal growth conditions were determined to be: temperature 37 degrees C, pH 7.0, inoculum size 0.1% and shaker rotary speed 250 r/min. Under the optimal conditions, the degradation kinetics of 2,6-DTBP with an initial concentration of 100 mg/L was studied. Results indicated that 62.4% of 2,6-DTBP was removed after 11 d. The degradation kinetics could be expressed by Eckenfelder equation with a half life of 9.38 d. In addition, the initial concentration of 2,6-DTBP played an important role on the degradation ability of the strain. The maximum initial concentration of 2,6-DTBP was determined to be 200 mg/L. Above this level, the strain was overloaded and exhibited significant inhibition.

The Isolation and Purification of Archaerhodopsin from Halobacterium sp. XZ515.

The claret membrane, instead of the purple membrane, was isolated when the conventional method for isolating purple membrane from Halobacterium halobium was applied to Halobacterium sp. XZ515. The SDS-PAGE results showed that only one protein, archaerhodopsin, existed in claret membrane with M. W. similar to BR. The method for isolating and purifying the archaerhodopsin, an intrinsic membrane protein, by detergent dissolution and hydrophobic chromatography on the column of octylsepharose, was also introduced. The pure archaerhodopsin showed the absorption spectrum similar to BR and was able to produce the M412 photoproduct in the photocycle under illumilation. It was concluded that this archaerhodopsin was a BR-like retinal protein.

[Research of hyperspectral reconstruction based on HJ1A-CCD data].

By restoring plentiful spectral information from several bands, hyperspectral image reconstruction could provide more suitable data source to water environment remote sensing. This is significant for inland water color remote sensing. By using the HJ1 A-HSI and HJ1A-CCD image acquired on June 6th, 2009, the hyperspectral data was reconstructed from HJ1A-CCD data. The results show that: (1) The average relative error of HJ1A-HSI data and reconstructed data compared with measured Rrs at 660 nm-900 nm are 0.333 5 and 0.307 7, respectively; (2) The entropy and average gradient of reconstructed image are higher than HJ1A-HSI image. In additional, the three band model get higher accuracy when inversing chl-a concentration by the reconstructed data.

[The degradation characteristics of degrading bacterium of 2,6-di-tert-butylphenol].

A degrading bacterial strain F-3-4 for 2,6-Di-tert-butylphenol (2,6-DTBP) was isolated from biofilm in acrylic fiber wastewater treatment structures. By acclimation, its capacity for degradation of 2,6-DTBP was enhanced by 26%. It was identified as Alcaligenes sp. according to morphological, physiological and biochemical characters. By tests in shaking flasks, the effects of the conditions of growth was studied, and it was determined that the optimum conditions of growth for the strain was 37 degrees C, pH 7.0 and inoculum amount 0.1%. Under these conditions, the kinetics of degradation for 2,6-DTBP of initial concentration 100 mg/L was studied, and the result indicated that the removal rate of 2,6-DTBP within 11 days was 62.4%, and the degradation process followed Eckenfelder kinetics. The half life of 2,6-DTBP was 9.38 days. The effect of initial concentration on degradation ability of the strain also was investigated. The results showed that the optimum initial concentration was 200 mg/L. When the initial concentrations were below it, the growth of strain and removal of 2,6-DTBP increased with the increase of initial concentration, while when above the value, they were inhibited.