Validation of an HPLC method for sildenafil citrate analysis in human plasma samples.

A simple, precise, rapid and accurate HPLC method for the determination of sildenafil citrate (SLD) in human plasma has been developed based on previous reports, to offer an alternative way to detect and quantify SLD and also to improve some characteristics of the validation process. Chromatography was carried out on a C18 reversed-phase column, using a mixture of acetonitrile: ammonium acetate (0.3 M pH 6.8) (1:1 v/v) as the mobile phase at a flow of 0.750 mL/min. Diazepam was used as an internal standard (IS) and detection was by UV at 240 nm. Samples were extracted with 200 microl of 0.1 M Na2CO3 and 5 mL of diethyl ether: dichloromethane (60:40). Retention times of SLD and IS were 4.8 and 7.1 min, respectively; total run time was 10 min. The linear range of SLD was found to be 20 - 1000 ng/mL. Limit of quantitation (LOQ) and limit of detection (LOD) were calculated to be 10 and 20 ng/mL, respectively. An improved percentage recovery of analyte is reported, showing a fast and reproducible approach to detect SLD in plasma human sample. The method was validated for its linearity, precision, accuracy, recovery, stability and specificity.

Effectiveness of prolonged low dose recombinant tissue-type plasminogen activator for refractory unstable angina.

OBJECTIVES: The aim of the present study was to evaluate the effectiveness of prolonged administration of thrombolytic therapy with low doses of recombinant tissue-type plasminogen activator (rt-PA) in patients with refractory unstable angina. BACKGROUND: Intracoronary thrombosis is often the cause of instability in patients with unstable angina. Thrombolytic therapy has been tested in these patients with conflicting results. METHODS: Sixty-seven patients with unstable angina refractory to standard antianginal therapy were randomized to receive, in addition to the common antianginal therapy, either rt-PA (0.03 mg/kg body weight per h for 3 consecutive days) plus heparin (to achieve activated clotting time of 250 to 400 s) (36 patients, group A) or the same dose of heparin plus placebo (31 patients, group B). RESULTS: No major bleeding was observed in either group of patients. One patient in group A and four in group B (2.7% vs. 12.9%, p < 0.01) developed acute myocardial infarction during the hospital period. Eight patients in group B underwent emergency coronary artery surgery or angioplasty because of worsening of symptoms. Group A patients had a significant reduction in the occurrence of chest pain compared with those in group B (95% confidence interval -7.2 to -2.1 episodes/3 days, p < 0.01). Patients in group B had a greater number of episodes of transient myocardial ischemia (237 vs. 103, p < 0.01) and a longer total ischemic burden (114 +/- 23 vs. 45.6 +/- 8.9 min/day, p < 0.01) than group A patients. After a mean follow-up of 14 +/- 6 months, group A patients were more frequently angina free and had a lower incidence of readmission to the hospital than group B patients. CONCLUSIONS: The combination of heparin and protracted administration of rt-PA at low doses is effective in stabilizing and reducing in-hospital adverse events in patients with unstable angina refractory to antianginal therapy.

[Anatomo-pathologic findings in 2 cases of Burkitt's lymphoma of the jaw]. / Rilievi anatomo-patologici su due casi di linfoma di Burkitt a sede mascellare.

The various types of Burkitt's lymphomas are classified with a review of their main clinical and histological features and particular reference to the Middle Eastern form.

Anti-inflammatory and antitumoural effects of Uncaria guianensis bark.

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria guianensis (Aublet) Gmell (Rubiaceae) is a medicinal plant from the jungles of South and Central America, used to treat cancer, arthritis, diabetes, and inflammation. Evaluate the anti-inflammatory and anti-tumor effects of Uncaria guianensis preparations. MATERIALS AND METHODS: Bio-guided fractionation of a hydroethanolic extract of Uncaria guianensis was performed, evaluating the fractions and subfractions for their effect on inflammatory mediators, tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and prostaglandin E2 (PGE2) by ELISA and nitric oxide (NO) by the Griess reaction in cultured supernatant from RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). The expression of cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS) and inhibitor of κB (IκB) were investigated in RAW 264.7 macrophages by flow cytometry. The activity of NF-κB in HeLa cells transfected with a luciferase reporter system was determined. The effect of Uncaria guianensis on the inflammatory response in vivo was assessed in BALB/c mice stimulated with LPS, on rat paw oedema induced by carrageenan, and on tumour growth and lung metastasis in BALB/c mice inoculated with 4T1 mammary tumour cells. Immune cell infiltrates and inflammatory mediators were evaluated in the tumour by immunohistochemistry. RESULTS: Sub-fraction Ug AIV inhibited, to varying degrees, NO, TNF-α, IL-6 and PGE2 production by macrophages in vitro (30 µg/ml) and in the serum of LPS-challenged mice (5 mg/kg). Macrophage expression of Cox-2 was inhibited (35%), IκB degradation was completely inhibited and NF-κB activation was inhibited (70%) by Ug AIV at 30 µg/ml. Ug AIV decreased paw oedema by 86% (5 mg/kg) and serum NO and TNF-α by 45% and 65% respectively. Ug AIV reduced 4T1 mammary tumour growth by 91% on day 33 post-inoculation as well as the levels of serum NO, IL-6 and TNF-α in the same animals. Ug AIV decreased the number of tumour-infiltrating T lymphocytes, macrophages and neutrophils as well as the number of cells positive for COX-2, iNOS, IL-6, TNF-α and p65. CONCLUSIONS: As Ug AIV was not cytotoxic for tumour cells or macrophages, its anti-tumour effect may be due to a reduction in pro-tumoural inflammatory processes in the tumour microenvironment, possibly mediated through NF-κB.