Aurora de la Peña

Reduction of no-reflow and reperfusion injury with the synthetic 17β-aminoestrogen compound Prolame is associated with PI3K/Akt/eNOS signaling cascade.

A high proportion of primary percutaneous coronary interventions performed in the setting of acute myocardial infarction, concur with inadequate myocardial perfusion at the microvascular level. This phenomenon, known as “no-reflow” contributes to reperfusion injury, poor prognosis and to unfavorable clinical outcome. In this study, we evaluated the hypothesis that the synthetic 17β-aminoestrogen Prolame, may confer cardioprotection and prevent against no-reflow. In an open-chest model of 30-min ischemia and 90-min reperfusion, male Wistar rats were randomly assigned to different groups: Control, Prolame, Prolame followed by the nitric oxide synthase inhibitor (l-NAME), and 17β-estradiol. Areas of risk, infarct size and no-reflow were determined by planimetry with triphenyltetrazolium chloride and thioflavin-S stains. Structural damage of the vasculature was measured as capillary compression in clarified tissue after intra-atrial injection of Microfil. Hemodynamic function was obtained at the end of stabilization, ischemia and reperfusion; nitric oxide (NO·) content was determined indirectly using the Griess reaction. Activation of the eNOS signaling cascade was determined by western blot. Prolame reduced the infarcted area, decreased the zones of no-reflow and capillary compression by activating the PI3K/Akt/eNOS signaling pathway in correlation with NO· increase. Prolame also activated endothelial cells augmenting NO· production, which was inhibited by ICI182780 (a selective estrogen receptor down-regulator), supporting the notion that the cardioprotective effect of Prolame involves the preservation of endothelium through the activation of estrogen receptor downstream signaling. Our results provide evidence that Prolame has potential therapeutic application in patients with AMI, as it prevents from both vascular and cardiac tissue damage.

Synthesis and molecular structure of prolame, N-(3-hydroxy-1,3,5(10)-estratrien-17β-yl)-3-hydroxypropylamine; an amino-estrogen with prolonged anticoagulant and brief estrogenic effects

The synthesis and molecular structure of prolame, N-(3-hydroxy-1,3,5(10)-estratrien-17 beta-yl)-3-hydroxypropylamine, is described. It was characterized by ir, nmr, mass spectrometry and chemical analysis. The crystal structure of this compound was determined by single-crystal x-ray diffraction. Prolame belongs to space group P212121. Cell dimensions are: a = 8.356(2), b = 13.343(4) and c = 16.119(4) A. Z = 4; R = 4.1%.

Comparative effect of synthetic aminoestrogens with estradiol on platelet aggregation

The in vitro effect upon platelet aggregation of estradiol and synthetic estrogens (prolame, buame, and proacame) is described. Prolame and buame produced a dose-dependent inhibition on platelet aggregation. Estradiol and proacame did not show anti-aggregating effects. The results suggest that the use of prolame and buame in estrogen therapy could reduce the risk of thrombo-embolic accidents.

Increased Fibrin Polymerization Rate in Patients with Primary Antiphospholipid Syndrome and Systemic Lupus Erythematosus

The main event in blood coagulation is the thrombincatalyzed conversion of fibrinogen into fibrin. This singular transformation of a soluble protein into an insoluble polymeric network occurs with faultless precision. Abnormalities of fibrin polymerization can lead to hemorragic and thrombotic disorders. Increased fibrinogen plasma concentration (Fg) and fibrin polymerization rate (FPR) could be additional risk factors associated with atherothrombosis in antiphospholipid syndrome (APS) and in systemic lupus erythematosus (SLE). Our objective was to investigate Fg and FPR in consecutive patients with APS and SLE. Thirty-nine patients and 31 ageand gender-matched healthy controls were studied. Sixteen patients had primary APS, 13 patients had SLE, and 10 patients had SLE plus APS. The mean of the FPR was significantly increased (0.2799 ± 0.091) in patients with APS plus SLE as compared with the control group (0.2052 ± 0.055) (p<0.05). Fg was higher in APS plus SLE (3.15 g/L ± 0.43) and in primary APS (3.03 g/L ± 0.29) than in controls (2.87 g/L ± 0.49). Our results demonstrated an increased FPR in patients with APS plus SLE. This phenomenon could be an additional risk factor for thrombosis in these autoimmune diseases.

The anticoagulant effect of prolame, N-(3-hydroxy-1,3,5(10) estratrien-17β-yl)-3-hydroxypropylamine, a novel amino-estrogen

The anticoagulant and estrogenic effects of prolame, N-(3-hydroxy-1,3,5(10)-estratrien-17 beta-yl)-3-hydroxypropylamine, are described. A single subcutaneous injection of prolame in male mice, ovariectomized mice, adult and infant male rats, produced dose-dependent increases of blood clotting time, which could be observed with the larger doses even after 4 days. In ovariectomized mice, prolame produced vaginal cornifications of shorter duration than those produced by estradiol-17 beta. The evidence suggests that, in contrast with currently used estrogens, prolame would not generate cardiovascular accidents if used for the treatment of prostatic carcinoma; it could also be exceptionally effective for the prevention of thrombosis.

The antithrombotic effect of the aminoestrogen prolame (N-(3-hydroxy-1,3,5(10)-estratrien-17B-YL)-3-hydroxypropylamine) is linked to an increase in nitric oxide production by platelets and endothelial cells

OBJECTIVE Women under hormone replacement therapy carry an increased risk of venous thromboembolism (VTE), mostly during the first year. Despite great efforts devoted to hormone therapy research, VTE remains a major drawback of estrogenic therapy, and the search for new compounds continues. We have synthesized and evaluated prolame, an aminoestrogen with anticoagulant properties. The aim of our work was to elucidate the anticoagulant mechanism of prolame. METHODS We studied the effects of prolame on nitric oxide (NO) synthesis in cultured endothelial cells and platelets using flow cytometry, on NO metabolites using a modified Griess method, on NO formation in vivo using electron paramagnetic resonance spectroscopy, on participation of nuclear estrogen receptors using flow cytometry, and on endothelial NO synthase (eNOS) mRNA expression using RT-PCR. We also studied the impact of prolame-treated endothelial cells (EC) on ADP-induced platelet aggregation, as well as the ability to prevent occlusive thrombi in an in vivo mice thrombosis model. RESULTS (a) Prolame induces NO production in ECs, platelets, and in a mouse model in vivo. (b) The NO-elevating effect of prolame can only be partially attributed to the nuclear estrogen receptors (ERs) since endothelial nitric oxide synthase (e-NOS) is slightly induced (37%) in ECs treated with prolame. (c) Platelets become 60% less responsive to aggregation induced by 10muM ADP when in suspension with prolame-treated ECs. (d) Prolame reduces the formation of thrombi in an in vivo thrombosis model. CONCLUSIONS Prolame could be a preferred alternative to other estrogens because of its reduced thromboembolic risk.

Intracellular Ca2+ stimulates the binding to androgen receptors in platelets

In this study, we demonstrated that ADP-induced platelet aggregation activates the binding of testosterone (T) to its receptor. It is well known that binding of ADP to its receptors induced the release of Ca2+ ions from dense bodies into the cytosol of platelets. In this work, we compared the binding of testosterone or dihydrotestosterone to their receptors using cytosol obtained from ADP-treated and non-treated platelets. These experiments were repeated using EGTA (a calcium chelator) or U73122 (a phospholipase C enzymatic activity inhibitor) to the ADP-treated platelets. In addition, we also developed a competition analysis for the androgen receptors (AR) using [3H]DHT, non-radioactive T, DHT or cyproterone acetate from ADP-treated platelets cytosol. The results from this study indicate that the cytosol obtained from non-ADP-treated platelets did not show any binding to [3H]T or [3H]DHT, whereas cytosol from ADP-treated platelets binds to the radio-labeled androgens. Furthermore cytosol from ADP plus U73122-treated platelets did not show binding to [3H]T or [3H]DHT. These data suggest that intracellular Ca2+ ions stimulates the binding of androgens to their receptors in platelets cytosol. The competition analysis shows that T and DHT have high affinities for the androgen receptors with similar IC50 values, whereas cyproterone acetate shows a lower affinity. The results from these data clearly indicate the presence of androgen receptors in platelets.

Palladium(II) and platinum(II) dichloro complexes containing diamine-estrone derivatives

The preparation ofcis-dichloro Pt(II) andcis-dichloro Pd(II) complexes ofN-[3-hydroxyestra 1:3:5 (10)trien-17β]ethylendiamine,N-[3-hydroxyestra 1:3:5 (10)trien-17β]1,3-propylendiamine, andN-[3-hydroxyestra 1:3:5 (10)trien-17β]2-aminomethylpyridine is reported. The complexes have been characterized by chemical analysis, infrared spectroscopy and molar conductivity.ZusammenfassungEs wird über die Darstellung voncis-Dichlor-Pt(II)- undcis-Dichlor-Pd(II)-Komplexen mitN-[3-Hydroxyestra 1:3:5 (10)trien-17β]ethylendiamin,N-[3-Hydroxyestra 1:3:5 (10) trien-17β]1,3-propylendiamin undN-[3-Hydroxyestra 1:3:5 (10)trien-17β]2-aminomethylpyridin berichtet. Die Komplexe wurden mittels chemischer Analyse, IR und Leitfähigkeitsmessungen charakterisiert.

Circulating Levels of Urokinase-Type Plasminogen Activator Receptor and D-Dimer in Patients With Hematological Malignancies

BACKGROUND Patients with cancer exhibit changes in their hemostatic mechanisms. The D-dimer (D-D) is the most important subproduct of fibrinolysis, and urokinase plasminogen activator receptor (uPAR) is related to invasiveness and metastases, and is overexpressed in neoplastic cells. The objective of this study was to identify in patients with hematological neoplasia, the serum levels of uPAR and D-D, and to determine their effects on outcome. PATIENTS AND METHODS A cross-sectional study was performed. Clinical and demographic data were obtained from the clinical chart. Determination of uPAR in serum (pg/L) was performed using an enzyme-linked immunosorbent assay, and D-D (μg/dL) using nephelometry. RESULTS We included 42 patients (35 with lymphomas). Statistically significant differences were found in D-D (P < .001) and uPAR (P < .01) between patients and control participants. Response was an accumulated clinical outcome. We observed statistical differences between groups (P < .001). D-D was positive in 70% of cases. CONCLUSION We found differences in D-D serum levels and soluble uPAR between control participants and patients with lymphoma. These results indicate that D-D serum levels and soluble uPAR should be considered biomarkers of response and survival in patients with lymphoma.