Endogenous Urotensin II Selectively Modulates Erectile Function through eNOS

Urotensin II (U-II) is a cyclic peptide originally isolated from the neurosecretory system of the teleost fish and subsequently found in other species, including man. U-II was identified as the natural ligand of a G-protein coupled receptor, namely UT receptor. U-II and UT receptor are expressed in a variety of peripheral organs and especially in cardiovascular tissue. Recent evidence indicates the involvement of U-II/UT pathway in penile function in human, but the molecular mechanism is still unclear. On these bases the aim of this study is to investigate the mechanism(s) of U-II-induced relaxation in human corpus cavernosum and its relationship with L-arginine/Nitric oxide (NO) pathway.Human corpus cavernosum tissue was obtained following in male-to-female transsexuals undergoing surgical procedure for sex reassignment. Quantitative RT-PCR clearly demonstrated the U-II expression in human corpus cavernosum. U-II (0.1 nM-10 µM) challenge in human corpus cavernosum induced a significant increase in NO production as revealed by fluorometric analysis. NO generation was coupled to a marked increase in the ratio eNOS phosphorilated/eNOS as determined by western blot analysis. A functional study in human corpus cavernosum strips was performed to asses eNOS involvement in U-II-induced relaxation by using a pharmacological modulation. Pre-treatment with both wortmannin or geldanamycinin (inhibitors of eNOS phosphorylation and heath shock protein 90 recruitment, respectively) significantly reduced U-II-induced relaxation (0.1 nM-10 µM) in human corpus cavernosum strips. Finally, a co-immunoprecipitation study demonstrated that UT receptor and eNOS co-immunoprecipitate following U-II challenge of human corpus cavernosum tissue.U-II is endogenously synthesized and locally released in human corpus cavernosum. U-II elicited penile erection through eNOS activation. Thus, U-II/UT pathway may represent a novel therapeutical target in erectile dysfunction

Development of a Headspace-Solid Phase Microextraction-Gas Chromatography Method to Determine Organohalogen Contamination in Drinking Water

The formation of organohalogen compounds in waters treated by chlorination has drawn increasing scientific attention due to the potentially hazardous health effects of this class of substances. Today, chlorination is the most widely used technology for civil water disinfection. In this study, headspace-solid phase microextraction coupled with GC-electron capture detector was used to determine organohalogen compounds in drinking water sampled from aqueducts and artesian wells in Italy. Experimental parameters, such as sample volume, stirring, salting out, extraction temperature, and extraction time, were evaluated and optimized. The LODs ranged from 1 to 10 ng/L and LOQs from 5 to 50 ng/L. A linear response was confirmed by correlation coefficients ranging from 0.9443 to 0.9999. Quantifiable organohalogen residues were found in 11 water samples, with concentration up to 11.3 +/- 0.5 mu g/L for the sum of all trihalomethanes and 0.66 +/- 0.03 mu g/L for the sum of trichloroethylene and tetrachloroethylene. These concentrations are lower than the current regulatory limits in Italy

Evaluation of flavonoids and furocoumarins in bergamot derivatives by HPLC-DAD

The high concentration of bergamot polyphenolic fraction is generating growing interest in all its derivatives due to the potential beneficial effects on human health, suggesting new application in the functional foods field. Once widely used in perfumery for the exclusive fragrance of its essential oil, the bergamot has reported, in the last decades, a drop in demand due to the synthetic essence placing on the market. In order to re-launch this citrus on the international market, the dry juice extract seems to be particularly suitable to be mixed with other foods with minimal impact on their organoleptic properties. The characterization of bergamot derivatives (juice, pastazzo and dry juice extract) was performed with HPLC-DAD method, with the detection and quantification of some flavonoids such as neohesperidin, naringin e neoeriocitrin, which represent an antioxidant pattern almost unique in nature, and of two furocoumarins, bergapten and bergamottin, potentially toxic if beyond certain limits. The method was validated in terms of detection limits (LODs), linearity, precision and accuracy

La qualità delle acque potabili in relazione allo stato di contaminazione causato da sostanze organoalogenate

In many drinking water treatment plants, the chlorination process is one of the main techniques used for water disinfection. This treatment leads to the formation of trihalomethanes and other halogenated compounds that draw significant scientific attention due to their potential adverse health effects on human beings. The Italian law (D.Lgs. 31/01, referring to EU directive 98/83) fixes restrictive limits (μg/L range) to regulate the concentration of disinfection by-products in drinking water. Considering these limits, in order to assure the drinking water quality, headspace solid-phase micro-extraction (HS-SPME) technique was applied for the organohalogenate compounds analysis. This method shows a good sensibility and precision for the determination of contaminant traces

Local cytokine production in patients with Acute Coronary Syndromes: a look into the eye of the perfect (cytokine) storm

none8openCirillo P; Cimmino G; D'Aiuto E; Di Palma V; Abbate G; Piscione F; Golino P; De Palma RCirillo, P; Cimmino, G; D'Aiuto, E; Di Palma, V; Abbate, G; Piscione, F; Golino, P; De Palma,

Quantitative RT-PCR for U-II in HCC.

<p>U-II is expressed as mRNA in HCC samples. Human tumoural cells SW-13 were used as positive control. Data were normalized on the basis of GAPDH and expressed as mean ± standard error of the mean (SEM) of three different human specimens.</p

Panel A: U-II induced concentration-response curve (0.1 nM–10 µM) in HCC strips was significantly inhibited by pre-treatment with wortmannin (0.1 µM), PI3K inhibitor, or geldanamycin (1 µM) Hsp90 inhibitor (***p<0.001 and ** p<0.01 <i>versus</i> vehicle).

<p>Data were expressed as the mean ± standard error of the mean (SEM) of six different specimens. Data were analyzed by ANOVA followed by Bonferroni post test. Panel B: NOx (total nitrite) production in HCC tissue incubated with U-II (1 nM-10 µM) or vehicle for 30 min. U-II caused a significant increase in NO production compared with vehicle (*<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001). Data were expressed as mean ± standard error of the mean (SEM) from four different specimens and analyzed by ANOVA followed by Bonferroni post test.</p

Co-immunoprecipitation analysis of UT receptor and eNOS.

<p>Tissues were stimulated with either vehicle (A) or U-II (C), lysates were incubated with mouse anti-eNOS. Lanes B and D correspond to the negative control of A and C, respectively. The western blot was probed with rabbit anti-GPR14 (UT receptor). U-II but not vehicle caused the co-immunoprecipation between eNOS and UT receptor.</p

Western blot analysis for eNOS and p-eNOS^Ser-1177 in HCC tissue.

<p>Panel A: U-II (10 µM) caused an increase in eNOS phosphorylation expressed as p-eNOS/eNOS ratio in a time-dependent manner (*p<0.05 <i>vs</i> vehicle). Panel B: U-II-induced eNOS phosphorylation (**p<0.01 <i>vs</i> vehicle), was significantly reverted by wortmannin (0.1 µM), PI3K inhibitor, (°p<0.05 <i>vs</i> U-II) but not by geldanamicin (1 µM), Hsp90 inhibitor. Data were expressed as the mean ± standard error of the mean (SEM) of four different specimens and were analyzed by ANOVA followed by Bonferroni post test.</p