Metabolic profiling of HepG2 cells incubated with S(−) and R(+) enantiomers of anti-coagulating drug warfarin

Warfarin is a commonly prescribed oral anticoagulant with narrow therapeutic index. It achieves anti-coagulating effects by interfering with the vitamin K cycle. Warfarin has two enantiomers, S(−) and R(+) and undergoes stereoselective metabolism, with the S(−) enantiomer being more effective. We reported the intracellular metabolic profile in HepG2 cells incubated with S(−) and R(+) warfarin by GCMS. Chemometric method PCA was applied to analyze the individual samples. A total of 80 metabolites which belong to different categories were identified. Two batches of experiments (with and without the presence of vitamin K) were designed. In samples incubated with S(−) and R(+) warfarin, glucuronic acid showed significantly decreased in cells incubated with R(+) warfarin but not in those incubated with S(−) warfarin. It may partially explain the lower bio-activity of R(+) warfarin. And arachidonic acid showed increased in cells incubated with S(−) warfarin but not in those incubated with R(+) warfarin. In addition, a number of small molecules involved in γ-glutamyl cycle displayed ratio variations. Intracellular glutathione detection further validated the results. Taken together, our findings provided molecular evidence on a comprehensive metabolic profile on warfarin-cell interaction which may shed new lights on future improvement of warfarin therapy

Optimization of conditions in random amplified polymorphic DNA (RAPD) reaction for determination of plant pathogenic bacteria

Random Amplified Polymorphic DNA (RAPD) technique is a powerful tool for genetic studies. It can be also applied for determination of bacterial species. The goal of this work was to optimize conditions for RAPD reaction, and to obtain RAPD patterns specific for some plant-associated bacteria. RAPD could be a good choice for screening and preliminary bacterial determination. Application of RAPD can determine that some of bacterial species are not reason of infection, and make additional tests for them unnecessary. This can provide time, labor, and as well as money saving, which is important particularly if expensive tests are applied. Optimization of reaction conditions in this work made possible good repeatability of RAPD electrophoretic patterns for particular bacterial species

Synthesis of 3a,12a-dihydroxy-23a,23b-dihomo-5b-cholan-24-oic acid

A novel multi-step synthesis of 3a,12a-dihydroxy-23a,23b-dihomo-5b-cholan-24-oic acid (23a,23b-dihomodeoxycholic acid) (5) has been achieved from methyl 3a,12a-dihydroxy-5b-cholanoate (1). Reduction of compound 1 with LiAlH4 in dry ether gave the corresponding alcohol 2 in 83 % yield. Selective esterification of compound 2 with tosyl chloride in dry piridine at 05 ºC for 12 h afforded the 3a,12a-dihydroxy-5b-24-cholanyl tosylate (3) in 64 % yield. The reaction of the tosyl derivative 3 with sodium diethyl malonate gave compound 4 which was first subjected to hydrolysis under basic conditions, followed by decarboxylation under acidic conditions to afford 3a,12a-dihydroxy-5b-23a,23b-dihomocholan-24-oic acid in 84 % yield

Sensory and color properties of dietary cookies with different fiber sources during 180 days of storage

The aim of this study was to examine the sensory properties of dietary cookies produced with addition of different dietary fiber sources (inulin and oligofructose, oat flakes, mixture of oat flakes and whole meal flour, whole meal flour and mixture of whole meal flour and carob flour), and to measure instrumentally color changes of cookies upper and lower surfaces, after storage at temperatures of 18-20 °C for 180 days. Addition of inulin and oligofructose was determined to be the fiber source with the highest impact on sensory properties. Storage for 180 days expressed the highest influence on texture properties of dietary cookies regardless of applied dietary fiber source. Color differences calculated from measured color properties (psychometric light, L*, psychometric tone, a*, and psychometric chrome, b*) between products including different fiber sources were noticeable to extremely noticeable among products, with the most expressed difference was registered for products containing carob flour.Cilj istraživanja je ispitivanje senzornih karakteristika dijetetskog keksa proizvedenog sa dodacima dijetetskih vlakana iz različitih izvora (inulin i oligofruktoza, ražene pahuljice, mešavina raženih pahuljica i integralnog brašna, integralno brašno i mešavina integralnog brašna i brašna rogača) i instrumentalno merenje promena boje gornje i donje površine keksa, korišćenjem "Minolta" Chroma meter CR 400 tristimulus colorimeter tokom skladištenja na temperaturi od 18-20 °C do 180 dana. Za dodatak inulina i oligofruktoze je utvrđeno da je izvor dijetetskih vlakana koji je najviše uticao na senzorna svojstva. Skladištenje od 180 dana imalo je najveći uticaj na teksturne promene dejetetskog keksa, nezavisno od izvora dijetetskih vlakana. Razlike u boji izračunate na bazi instrumentalno merenih svojstva boje proizvoda (L*, a*, b*) izmeću proizvoda različitog sastava u pogledu izvora dijetnih vlakana su bile izražene do ekstremno izražene, sa najupečatljivijim razlikama registrovanim u slučaju dodatka brašna od rogača