Novel reverse thia-analogs of fosmidomycin: Synthesis and antiplasmodial activity.

Thia analogs of fosmidomycin are potent inhibitors of the non-mevalonate isoprenoid biosynthesis enzyme 1-deoxy-d-xylulose 5-phosphate reductoisomerase (IspC, Dxr) of Plasmodium falciparum. Several new thioethers displayed antiplasmodial in vitro activity in the low nanomolar range, without apparent cytotoxic effects in HeLa cells. The (S)-(+)-enantiomer of a typical representative selectively inhibited IspC and the growth of P. falciparum in continuous culture. The inhibitor was stable at pH 7.6 and room temperature, and no racemization was observed under these conditions during a period of up to two days. Oxidation of selected thioethers to sulfones reduced antiplasmodial activity and the inhibitory activity against Escherichia coli, Mycobacterium tuberculosis and P. falciparum IspC orthologs.

IspC as target for antiinfective drug discovery: synthesis, enantiomeric separation, and structural biology of fosmidomycin thia isosters.

The emergence and spread of multidrug-resistant pathogens are widely believed to endanger human health. New drug targets and lead compounds exempt from cross-resistance with existing drugs are urgently needed. We report on the synthesis and properties of "reverse" thia analogs of fosmidomycin, which inhibit the first committed enzyme of a metabolic pathway that is essential for the causative agents of tuberculosis and malaria but is absent in the human host. Notably, IspC displays a high level of enantioselectivity for an α-substituted fosmidomycin derivative.

Reference intervals for plasma L-arginine and the L-arginine:asymmetric dimethylarginine ratio in the Framingham Offspring Cohort.

L-arginine, as a precursor of NO synthesis, has attracted much scientific attention in recent years. Experimental mouse models suggest that L-arginine supplementation can retard, halt, or even reverse atherogenesis. In human studies, supplementation with L-arginine improved endothelium-dependent vasodilation. However, L-arginine levels are best interpreted in the context of levels of asymmetric dimethylarginine (ADMA), a competitive inhibitor of NO synthase. Thus, reference limits for circulating L-arginine and the L-arginine:ADMA ratio may help to determine the nutritional state of individuals at high cardiovascular risk in light of increased ADMA levels. We defined reference limits for plasma L-arginine in 1141 people and for the L-arginine:ADMA ratio in 1138 relatively healthy individuals from the Framingham Offspring Cohort. Plasma L-arginine and ADMA concentrations were determined by using a stable isotope-based LC-MS/MS method. The reference limits (2.5th and 97.5th percentiles) for plasma L-arginine were 41.0 µmol/L (95% CI = 39.5-42.5 µmol/L) and 114 µmol/L (95% CI = 112-115 µmol/L), whereas corresponding reference limits (2.5th and 97.5th percentiles) for the L-arginine:ADMA ratio were 74.3 µmol/L (95% CI = 71.1-77.3 µmol/L) and 225 µmol/L (95% CI = 222-228 µmol/L). Plasma L-arginine was positively associated with the estimated glomerular filtration rate (eGFR) and blood glucose levels, whereas the L-arginine:ADMA ratio was positively associated with eGFR and diastolic blood pressure but inversely associated with homocysteine and (log)C-reactive protein. We report reference levels for plasma L-arginine and for the L-arginine:ADMA ratio that may be helpful for evaluation of the effects of L-arginine supplementation in participants with an impaired L-arginine/NO pathway.

Plasma symmetric dimethylarginine reference limits from the Framingham offspring cohort.

BACKGROUND: Symmetric dimethylarginine (SDMA) is a by-product of protein methylation. Once released from proteins, SDMA is eliminated by the kidneys; consequently, plasma concentration has been suggested as a sensitive marker of renal function. Furthermore, recent work implicates SDMA in the pathogenesis of cardiovascular disease. To date, reference limits for SDMA plasma concentrations in healthy individuals are lacking. METHODS: This study defined reference limits for plasma SDMA concentrations in 840 relatively healthy individuals of the Offspring Cohort from Framingham Heart Study (mean age 56 years, 61% women). Plasma SDMA concentrations were determined by LC-MS/MS using a stable isotope dilution assay. RESULTS: The median SDMA concentration in the reference sample was 0.37 µmol/L (Q1, Q3:0.32, 0.43 µmol/L) and the reference limits were 0.225 and 0.533 (2.5th and 97.5th percentile). In a multivariable regression model, serum creatinine, age and total homocysteine were positively associated with SDMA (p<0.001 for all), whereas the body mass index and diastolic blood pressure were inversely related to SDMA (p-values<0.01 and 0.03, respectively). CONCLUSIONS: This study reports plasma SDMA reference limits from the community-based Framingham Heart Study. Plasma SDMA concentration was related positively to advancing age, but inversely to renal function. These reference limits may allow the identification of individuals with raised plasma SDMA concentrations.

Asymmetric dimethylarginine reference intervals determined with liquid chromatography-tandem mass spectrometry: results from the Framingham offspring cohort.

BACKGROUND: Accumulating evidence links higher circulating asymmetric dimethylarginine (ADMA) to greater risk of cardiovascular disease (CVD). Relatively small differences in ADMA concentrations between healthy individuals and those with disease underscore the need to formulate reference intervals that may aid risk stratification of individuals. METHODS: We formulated reference intervals for plasma ADMA concentrations using a community-based reference sample from the Framingham Offspring Study consisting of 1126 nonsmoking individuals [mean (SD) age 56 (9) years; 60% women] who were free of clinical CVD, hypertension, diabetes, and obesity and who attended a routine examination at which ADMA was assayed. ADMA concentrations were determined using a validated tandem mass spectrometry-liquid chromatography assay. RESULTS: In the study sample, the mean ADMA concentration was 0.52 (0.11) micromol/L, and the reference limits were 0.311 and 0.732 (2.5th and 97.5th percentile). The sex-specific reference limits were 0.310 and 0.745 in men and 0.313 and 0.721 micromol/L in women. In multivariable regression analysis, ADMA plasma concentrations were positively correlated with age and total plasma homocysteine (both P < 0.001). CONCLUSIONS: Reference limits calculated for circulating ADMA in our large community-based healthy reference sample confirm the previous observation of a relatively narrow distribution of concentrations. This suggests a tight physiological control of ADMA plasma concentrations, presumably by dimethylarginine dimethylaminohydrolase (DDAH) metabolism of ADMA.

Acute effects of various fast-food meals on vascular function and cardiovascular disease risk markers: the Hamburg Burger Trial.

BACKGROUND: High-fat meals have negative effects on endothelial function, but vitamin-rich side orders may prevent these negative effects. OBJECTIVE: The acute effects of conventional and alternative fast-food meals on vascular function and various cardiovascular biomarkers were investigated. DESIGN: In a crossover study, flow-mediated endothelium-dependent dilatation (FMD) and cardiovascular disease risk markers were investigated in 24 healthy volunteers before and 2 and 4 h after 3 fast-food meals: a conventional beef burger with French fries, ketchup, and carbonated lemon-flavored soda (meal 1); a vegetarian burger with French fries, ketchup, and carbonated lemon-flavored soda (meal 2); and a vegetarian burger with salad, fruit, yogurt, and orange juice (meal 3). RESULTS: FMD decreased after all 3 fast-food meals: the values were 9.7 +/- 2.5%, 7.5 +/- 3.5%, and 6.2 +/- 3.3% for meal 1; 9.2 +/- 3.4%, 7.1 +/- 3.4%, and 6.3 +/- 4.0% for meal 2; and 8.8 +/- 3.3%, 6.2 +/- 4.0%, and 6.8 +/- 4.3% for meal 3 at baseline, 2 h, and 4 h, respectively. There were significant intraindividual differences for time (P < 0.001) but not for type of meal (P = 0.677). A postprandial increase in baseline diameter of the brachial artery was significant for time (P < 0.001) but not for type of meal (P = 0.148). CONCLUSIONS: Against common expectations, a conventional beef burger meal and presumably healthier alternatives with or without vitamin-rich side orders did not differ significantly in their acute effects on vascular reactivity. The frequently reported postprandial decline in FMD may be attributed in part to a postprandial increase in baseline arterial diameter.

High-throughput liquid chromatographic-tandem mass spectrometric determination of arginine and dimethylated arginine derivatives in human and mouse plasma.

The balance between nitric oxide (NO) and vasoconstrictors like endothelin is essential for vascular tone and endothelial function. L-Arginine is converted to NO and L-citrulline by NO synthase (NOS). Asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) are endogenous inhibitors of NO formation. ADMA is degraded by dimethylamino dimethylhydrolases (DDAHs), while SDMA is exclusively eliminated by the kidney. In the present article we report a LC-tandem MS method for the simultaneous determination of arginine, ADMA, and SDMA in plasma. This method is designed for high sample throughput of only 20-mul aliquots of human or mouse plasma. The analysis time is reduced to 1.6 min by LC-tandem MS electrospray ionisation (ESI) in the positive mode. The mean plasma levels of l-arginine, ADMA, and SDMA were 74+/-19 (SD), 0.46+/-0.09, and 0.37+/-0.07 microM in healthy humans (n=85), respectively, and 44+/-14, 0.72+/-0.23, and 0.19+/-0.06 microM in C57BL/6 mice. Also, the molar ratios of arginine to ADMA were different in man and mice, i.e. 166+/-50 and 85+/-22, respectively.