Stability of selected chlorinated thiazide diuretics.

In sports, diuretics are used for two main reasons: to flush previously taken prohibited substances with forced diuresis and in sports where weight classes are involved to achieve acute weight loss. A common property observed for thiazides is hydrolysis in aqueous media resulting in the formation of the degradation product aminobenzenedisulphonamide. This degradation product can be observed for several thiazides. Because there is limited information regarding the effect of pH, temperature and light on the stability of thiazides, these parameters were investigated for chlorothiaizide, hydrochlorothiazide and altizide. For all three compounds the degradation product could be detected after incubation at pH 9.5 for 48h at 60 degrees C. At lower pH and temperature the degradation product could not be detected for all compounds. When samples were exposed to UV-light altizide and hydrochlorothiazide were photodegraded to chlorothiazide. When the degradation rate between the different compounds was compared for a given temperature and pH, altizide is the most unstable compound. This study confirms that thiazide degradation products can be formed in urine during transport. Hence doping control laboratories shall include them into their routine testing methods as required by WADA.

Genomic and epidemiological monitoring of yellow fever virus transmission potential.

The yellow fever virus (YFV) epidemic in Brazil is the largest in decades. The recent discovery of YFV in Brazilian Aedes species mosquitos highlights a need to monitor the risk of reestablishment of urban YFV transmission in the Americas. We use a suite of epidemiological, spatial, and genomic approaches to characterize YFV transmission. We show that the age and sex distribution of human cases is characteristic of sylvatic transmission. Analysis of YFV cases combined with genomes generated locally reveals an early phase of sylvatic YFV transmission and spatial expansion toward previously YFV-free areas, followed by a rise in viral spillover to humans in late 2016. Our results establish a framework for monitoring YFV transmission in real time that will contribute to a global strategy to eliminate future YFV epidemics.

In vitro inhibition of platelet aggregation by bile salts.

In a series of in vitro experiments, the influence of bile salts on platelet aggregation by ADP or by collagen and on serotonin-14C release by collagen, was studied. Sodium salts of the following bile acids showed a clear inhibitory effect: glycochenodeoxycholic acid, taurochenodeoxycholic acid, glycocholic acid, taurocholic acid and cholic acid. Lowering the pH of the platelet-rich plasma resulted in decrease platelet aggregation by ADP and by collagen. Bile salts further enhanced the inhibitory effect of pH change. In contrast the sodium salt of chenodeoxycholic acid was the sole aggregation inducing bile salt we studied. Relating the above studies to the clinical situation of upper gastroduodenal mucosal haemorrhage, we suggest that biliary reflux as well as the acidic environment may contribute to a poor haemostatic response by impaired platelet aggregation in the upper gastrointestinal tract of otherwise normal human subjects.

Inhibitory effect of acetylsalicylic acid on human platelet function in normal volunteers and in women using a combined oral contraceptive regime.

Six parameters related to the release reaction were measured simultaneously in 10 human volunteers prior to the intake of one single dose of 1 g acetylsalicylic acid and 1, 4, 5, 6 and 7 days alter: deltaE5 with diluted collagen, deltaE10 with Thrombofax and the serotonin-14C release by undiluted and diluted collagen, by Thrombofax and by bovine plasma. The duration of the inhibitory effect varied according to the test used. It was the most prolonged (through the 7th day) if serotonin-14C release by diluted collagen was measured. A systematic investigation of the platelet release reaction in women taking a combined oral contraceptive was also performed. There were no statistically significant differences from a control group. No difference in acetylsalicylic acid sensitivity, measured 24 hours after intake of 1 g of aspirin, could be demonstrated.

Fibrinolysis and coagulation in patients with infectious disease and sepsis.

Sepsis is often associated with hemostatic dysfunction. This study aimed to relate changes in fibrinolysis and coagulation parameters to sepsis and sepsis outcome. Urokinase-type plasminogen activator (u-PA) antigen, tissue-type plasminogen activator (t-PA) antigen and activity, plasminogen activator inhibitor (PAI) type 1 antigen, PAI activity, antithrombin (AT) III activity, and protein C activity were measured in 24 patients suffering from sepsis or septic shock and the results were compared with those observed in 30 non-sepsis patients with severe infectious disease. The u-PA level was markedly increased in plasma of sepsis patients as compared to non-sepsis patients (11.5 +/- 9.4 versus 1.6 +/- 1.5 ng/ml, p less than 0.0001). PAI-1 antigen and t-PA activity showed a significant increase in sepsis patients (320 +/- 390 ng/ml versus 120 +/- 200 ng/ml, and 3.0 +/- 3.6 IU/ml versus 1.0 +/- 0.7 IU/ml, respectively, p less than 0.01). AT III was decreased in sepsis patients (58 +/- 28% in sepsis versus 79 +/- 26% in severe infectious disease, p less than 0.01) as was protein C (30 +/- 18% versus 58 +/- 27%, p less than 0.001). No significant difference was found for t-PA antigen nor for PAI activity. Nonsurvivors of sepsis were distinguished mainly by a high u-PA antigen level and increased t-PA activity. It is concluded that plasma u-PA antigen showed the strongest significant difference, among the parameters evaluated, between sepsis and severe infection. u-PA antigen may be of prognostic value in patients admitted to the medical intensive care unit for severe infectious disease.

Granulocyte elastase, tumor necrosis factor-alpha and urokinase levels as prognostic markers in severe infection.

We have examined the prognostic value of the levels in the blood of granulocyte elastase-alpha 1-proteinase inhibitor (E-alpha 1-PI) complex, tumor necrosis factor-alpha (TNF-alpha) and urokinase-type plasminogen activator (u-PA) in 35 patients with severe infection upon admission to an Intensive Care Unit. Fourteen patients died. No differences for E-alpha 1-PI complex were found between survivors and nonsurvivors, but in all patients the levels on admission were eight-fold higher than the reference value. TNF-alpha levels, measured by immunoassay, on admission were four times higher in the nonsurvivors than in the survivors (p = 0.0003) and correlated with the severity of the disease (APACHE II score, r = 0.43, p less than 0.05). TNF-alpha was not detectable by bioassay. Total u-PA antigen (u-PA Ag), plasmin-activatable single-chain u-PA (scu-PA) and inactive, nonactivatable u-PA (u-PA#) were on admission all two-fold higher in the nonsurvivors (p = 0.0006, 0.003 and 0.0003, respectively), while normal in the survivors. In both, survivors and nonsurvivors, the ratio between scu-PA and u-PA Ag was significantly decreased (p less than 0.001, compared to a reference group of healthy volunteers), indicative for enhanced conversion of scu-PA to active two-chain u-PA (tcu-PA) and inactive u-PA# during severe infectious disease. tcu-PA was detected in nine of the 35 patients, while virtually undetectable in controls. scu-PA correlated with the Child-Pugh score on admission (r = 0.42, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Intra-individual variability of fibrinogen levels.

Elevated fibrinogen concentrations are recognized as playing an important role in the pathogenesis of atherosclerosis. In the framework of a risk factor survey in 342 middle-aged working men, screened twice over a period of five months, plasma fibrinogen levels were found to be fairly unstable as large discrepancies between both measurements were observed. Due to a substantial proportion of within-person variability, the reliability coefficient was only R = 0.56. Repeatability was highest in higher educated and physically more active men. Our data suggest that the impact of elevated fibrinogen levels on the development of ischemic heart disease and stroke, is likely to be under-estimated.

Relation of fibrinogen to lifestyles and to cardiovascular risk factors in a working population.

BACKGROUND: The association between fibrinogen and smoking behaviour, age, body mass index, blood pressure, heart rate and plasma lipid profile, was assessed in a cohort of middle-aged working men. METHODS: Seven hundred and forty five subjects were examined as part of a health intervention programme at the work-site. Nine subjects were excluded from the study because of prevalent diabetes. Correlates of plasma fibrinogen concentrations were evaluated through univariate and multivariate methods. RESULTS: In multiple regression analysis fibrinogen correlated with age, smoking behaviour, apolipoprotein B (apo B) apolipoprotein A-I (apo A-I) and lipoprotein (a) (Lp(a)) levels, which together explained 11% of the variation in fibrinogen concentration. From this model, fibrinogen concentration was associated with an increase of 13.6 mg/dl for every 10 years' increase in age, 28.2 mg/dl if a person smoked, and 4.6 mg/dl and 3.8 mg/dl with a 20 mg/dl rise in respectively apo B and Lp(a). A 20 mg/dl increase in apo A-I concentrations was estimated to be associated with a 6.0 mg/dl lower fibrinogen level. CONCLUSION: The data indicate that both biochemical and lifestyle factors are related to the plasma fibrinogen concentration; these interactions may explain partly the relationship between fibrinogen and cardiovascular disease.

Bromocriptine in an injectable retard form for puerperal lactation suppression: comparison with Estandron prolongatum.

Intramuscular injection of a single 50-mg dose of long-acting bromocriptine microspheres was compared with a single intramuscular dose of an estradiol/testosterone ester combination in a single-blind, randomized study of 54 subjects. Bromocriptine was significantly more effective than the steroid drug in preventing milk flow, and rebound lactation was not observed in any bromocriptine-treated patients. Neither group showed deleterious side effects or significant biologic changes in coagulation parameters. There were no blood pressure or electrocardiographic alterations. Postpartum prolactin suppression was more intense after bromocriptine administration than after steroid therapy.

Interpretation of urinary concentrations of pseudoephedrine and its metabolite cathine in relation to doping control.

Until the end of 2003 a urinary concentration of pseudoephedrine exceeding 25 microg/mL was regarded as a doping violation by the World Anti-Doping Agency. Since its removal from the prohibited list in 2004 the number of urine samples in which pseudoephedrine was detected in our laboratory increased substantially. Analysis of 116 in-competition samples containing pseudoephedrine in 2007 and 2008, revealed that 66% of these samples had a concentration of pseudoephedrine above 25 microg/mL. This corresponded to 1.4% of all tested in competition samples in that period. In the period 2001-2003 only 0.18% of all analysed in competition samples contained more than 25 microg/mL. Statistical comparison of the two periods showed that after the removal of pseudoephedrine from the list its use increased significantly. Of the individual sports compared between the two periods, only cycling is shown to yield a significant increase.Analysis of excretion urine samples after administration of a therapeutic daily dose (240 mg pseudoephedrine) in one administration showed that the threshold of 25 microg/mL can be exceeded. The same samples were also analysed for cathine, which has currently a threshold of 5 microg/mL on the prohibited list. The maximum urinary concentration of cathine also exceeded the threshold for some volunteers. Comparison of the measured cathine and pseudoephedrine concentrations only indicated a poor correlation between them. Hence, cathine is not a good indicator to control pseudopehedrine intake. To control the (ab)use of ephedrines in sports it is recommended that WADA reintroduce a threshold for pseudoephedrine.

Verworven Hypofibrinogenemie.

The pathogenesis of acquired hypofibrinogenemia has been investigated in 52 cases by several blood coagulation parameters and by clinical and pathological findings. Disseminated intravascular coagulation, fibrinolysis, deficiency of synthesis and combinations of decreased synthesis with disseminated intravascular coagulation or fibrinolysis were found. The pathogenesis of acquired hypofibrinogenemia and its treatment are discussed.

Antihaemophilic factor A activity, F VIII-related antigen and von Wilebrand factor in hepatic cirrhosis.

F VIII activity, F VIII-related antigen and von Willebrand factor were measured in 46 patients with hepatic cirrhosis and in 30 normal individuals. These parameters were significantly higher in hepatic cirrhosis than in the controls. Linear relationships between F VIII activity and F VIII-related antigen and between F VIII-related antigen and von Willebrand factor were found in patients with hepatic cirrhosis as well as in normal individuals. However, in both groups no relationship between F VIII activity and von Willebrand factor was present. The existence of a low-grade intravascular coagulation in hepatic cirrhosis may be postulated but more information about the metabolism of F VIII protein is needed before such a statement can be proven.

Identification of two different mutations causing protein S deficiency in two unrelated Belgian families using a nonisotopic scanning and sequencing method.

Hereditary protein S deficiency is a risk factor for developing recurrent venous thromboembolic disease and is caused by a defect in the protein S 1 (PROS1) gene. Identification of the mutation in the PROS1 gene can overcome diagnostic uncertainty in family members with borderline protein S levels. We describe a novel nonisotopic method for molecular diagnosis of protein S deficiency, using fluorescein-labeled amplification and sequencing primers. As a first step, all exons of the PROS1 gene are selectively amplified, and heteroduplex analysis is performed. As a second step, all exons are analyzed by direct sequencing. Using this method, we have characterized the molecular defect in two Belgian families with hereditary protein S deficiency type I: a frameshift mutation in exon XIV (1881insTC) and a missense mutation caused by a T-to-C transition, resulting in substitution of Leu405 by Pro (L405P).

Effects of oxametacin on coumarin anticoagulation and on platelet function in humans.

In 12 chronically anticoagulated patients the administration of 1-p-chlorobenzoyl-5-methoxy-2-methyl-3-indoacetohydroxamic acid (oxametacin, Flogar), three times 100 mg a day, decreased the thrombotest percentage from a mean of 11.2% to a mean of 8.3% after one week and of 7.8% after two weeks of treatment. A potentiating effect of oxametacin was also found when the prothrombin time or the activated partial thromboplastin time were used as parameters. In one third of our patients an adaptation of the coumarin dose or even an interruption in the warfarin administration was necessary. Although no severe bleedings occurred, care should be taken in prescribing oxametacin to anticoagulated patients. In a second part of this study we compared the ex vivo effects of a single oral dose of 1 g of acetylsalicylic acid, 50 mg of indometacin and 100 mg of oxametacin in human volunteers. Platelet aggregation and 5HT--14C release induced by collagen. Thrombofax, ADP, adrenaline (epinephrine), bovine plasma and ristocetin were measured before, 1 and 24 h after drug administration. A clear-cut inhibitory effect as induced by acetylsalicylic acid was not found for oxametacin.