Expression Profiling of Nucleotide Metabolism-Related Genes in Human Breast Cancer Cells After Treatment with 5-Fluorouracil

5-Fluorouracil (5-FU) is one of the most widely used drugs for treatment of cancers, including breast cancer that exhibits its anticancer activity by inhibiting DNA synthesis and also incorporated into DNA and RNA. The objective of this investigation was to find out the total nucleotide metabolism genes regulated by 5-FU in breast cancer cell line. The breast cancer cell line MCF-7 was treated with the drug 5-FU. To analyze the expression of genes, we have conducted the experiment using 1.7k and 19k human microarray slide and confirmed the expression of genes by semiquantitative reverse transcription-polymerase chain reaction. The expression of 44 genes involved in the nucleotide metabolism pathway was quantified. Of these 44 genes analyzed, transcription of 6 genes were upregulated and 9 genes were downregulated. Earlier studies revealed that the transcription of genes for key enzymes like thymidylate synthase, thymidinekinase, and dihydropyrimidine dehydrogenase are regulated by 5-FU. This study identified some novel genes like thioredoxin reductase, ectonucleotide triphosphate dephosphorylase, and CTP synthase are regulated by 5-FU. The data also reveal large-scale perturbation in transcription of genes not involved directly in the known mechanism of action of 5-FU.

Standardization and validation of an induced ovulation model system in buffalo cows: Characterization of gene expression changes in the periovulatory follicle

In bovines characterization of biochemical and molecular determinants of the dominant follicle before and during different time intervals after gonadotrophin surge requires precise identification of the dominant follicle from a follicular wave. The objectives of the present study were to standardize an experimental model in buffalo cows for accurately identifying the dominant follicle of the first wave of follicular growth and characterize changes in follicular fluid hormone concentrations as well as expression patterns of various genes associated with the process of ovulation. From the day of estrus (day 0), animals were subjected to blood sampling and ultrasonography for monitoring circulating progesterone levels and follicular growth. On day 7 of the cycle, animals were administered a PGF2α analogue (Tiaprost Trometamol, 750 μg i.m.) followed by an injection of hCG (2000 IU i.m.) 36 h later. Circulating progesterone levels progressively increased from day 1 of the cycle to 2.26 ± 0.17 ng/ml on day 7 of the cycle, but declined significantly after PGF2α injection. A progressive increase in the size of the dominant follicle was observed by ultrasonography. The follicular fluid estradiol and progesterone concentrations in the dominant follicle were 600 ± 16.7 and 38 ± 7.6 ng/ml, respectively, before hCG injection and the concentration of estradiol decreased to 125.8 ± 25.26 ng/ml, but concentration of progesterone increased to 195 ± 24.6 ng/ml, 24 h post-hCG injection. Inh-α and Cyp19A1 expressions in granulosa cells were maximal in the dominant follicle and declined in response to hCG treatment. Progesterone receptor, oxytocin and cycloxygenase-2 expressions in granulosa cells, regarded as markers of ovulation, were maximal at 24 h post-hCG. The expressions of genes belonging to the super family of proteases were also examined; Cathepsin L expression decreased, while ADAMTS 3 and 5 expressions increased 24 h post-hCG treatment. The results of the current study indicate that sequential treatments of PGF2α and hCG during early estrous cycle in the buffalo cow leads to follicular growth that culminates in ovulation. The model system reported in the present study would be valuable for examining temporo-spatial changes in the periovulatory follicle immediately before and after the onset of gonadotrophin surge.

Structure of Disodium Deoxyeytidine 5'-Phosphate Heptahydrate, C9H 12N30TP2-.2N a +. 7H 20

M,=477.3, orthorhombic, P2~2~2~, a= 6.719.(4), b=29.614(15), c= 9.559 (3) ~, Z=4, U-- 1902.0 A 3, D x = 1.67 Mg m -3, 2(Cu Ka) = 1.5418A, /~=l.90mm -1, T=290K. Final R for 1809 observed reflections is 0.045. The structure shows an unusual gauche-trans conformation about the C(4')-C(5') bond, while the sugar pucker [C(3')-exo] and glycosidic torsion angle [)CCN = 70.2 (5) °, anti] are normal. The two Na + ions do not interact with the molecule directly, being completely surrounded by water molecules. The cytosine bases are stacked, with a separation distance of 3.36 (5) A.

5,7-Dimethoxy-2-phenyl-4H-chromen-4-one

The asymmetric unit of the title compound, C17H14O4, contains two independent molecules which differ in the relative orientations of the phenyl rings with repect to the essentially planar [maximum deviations of 0.029 (2) and 0.050 (2) angstrom in the two molecules] chromene fused-ring system, forming dihedral angles of 10.3 (5) and 30.86 (5)degrees in the two molecules. The crystal structure is stabilized by weak C-H center dot center dot center dot O and C-H center dot center dot center dot pi interactions, and pi-pi stacking interactions.

Chlorine NQR in 3-amino-2-chloropyridine and 2-chloro-5-nitropyridine

Temperature dependence of 35Cl nuclear quadrupole resonance (NQR) frequencies has been studied in 3-amino-2-chloropyridine and 2-chloro-5-nitropyridine from 77 to 298 K. The data were analysed and the torsional frequencies regarding internal motions in the molecules evaluated in the above temperature range using Bayer's theory and Brown's method.

Measurement of Turbulence and pitch of the airstream in the 5'* 7' tunnel of Indian Institute of Science

Abstract is not available.

Effects Of 5' Flanking Sequences And Changes In The 5' Internal Control Region On The Transcription Of Rice Transfer-Rna Gcc Cly Gene

A stretch of 71 nucleotides in a 1.2 kilobase pair Pst I fragment of rice DNA was identified as tRNA~ gene by hybridization and nucleotide sequence analyses. The hybridization of genomic DNA with the tRNA gene showed that there are about 10 glycine tRNA genes per diploid rice genome. The 3' and 5' internal control regions, where RNA polymerase III and transcription factors bind, were found to be present in the coding sequence. The gene was transcribed into a 4S product in an yeast cell-free extract. The substitution of 5' internal control region with analogous sequences from either M13mpl9 or M13mpl8 DNA did not affect the transcription of the gene in vitro. The changes in three highly conserved nucleotides in the consensus 5' internal control region (RGYNNARYGG; R = purine, Y = pyrimidine, N = any nucleotide) did not affect transcription showing that these nucleotides are not essential for promotion of transcription. There were two 16 base pair repeats, 'TGTTTGTTTCAGCTTA' at - 130 and - 375 positions upstream from the start of the gene. Deletion of 5' flanking sequences including the 16 base pair repeat at - 375 showed increased transcription indicating that these sequences negatively modulate the expression of the gene.

Evaluation Of Diffusion-Coefficient And Ionic Mobility In (Nh4)4fe(Cn)6.1.5 H2o

The diffusion coefficient, D, and the ionic mobility, μ, in the protonic conductor ammonium ferrocyanide hydrate have been determined by the isothermal transient ionic current method. D is also determined from the time dependence of the build up of potential across the samples and theretical expressions describing this build up in terms of double exponential dependence on time are obtained. The values obtained are D=3.875×10−11m2s−1 and μ=1.65×10−9 m2V−1s−1.

Structure of (6S,13bR)-1,2,3,5,6,13b-hexahydro-6-isopropyl-8H-pyrrolo[1',2':1,2]pyrazino[3,4,-b]quinazoline-5,8-dione

C17H19N302, monoclinic, P21, a = 5.382 (1), b = 17.534(4), c = 8.198(1)/L ,8 = 100.46(1) °, Z= 2, d,, = 1.323, dc= 1.299 Mg m-3, F(000) = 316, /~(Cu .Ka) = 0.618 mm -1. R = 0.052 for 1284 significant reflections. The proline-containing cispeptide unit which forms part of a six-membered ring deviates from perfect planarity. The torsion angle about the peptide bond is 3.0 (5) ° and the peptide bond length is 1.313 (5)A. The conformation of the proline ring is Cs-Cf~-endo. The crystal structure is stabilized by C-H... O interactions.

Interaction of Cibacron Blue F3G-A and Procion Red HE-3B with sheep liver 5,10-methylenetetrahydrofolate reductase

Cibacron Blue F3G-A, a probe used to monitor nucleotide binding domains in enzymes, inhibited sheep liver 5, 10-methylenetetrahydrofolate reductase competitively with respect to 5-methyltetrahydrofolate and NADPH. The Ki values obtained by kinetic methods and the Kd value for the binding of the dye to the enzyme estimated by protein fluorescence quenching were in the range 0·9-1·2 μM. Another triazine dye, Procion Red HE-3B interacted with the enzyme in an essentially similar manner to that observed with Cibacron Blue F3G-A. These results as well as the interaction of the dye with the enzyme monitored by difference spectroscopy and intrinsic protein fluorescence quenching methods indicated that the dye was probably interacting at the active site of the enzyme by binding at a hydrophobic region.

The Structure of 9b-Methyl- 3afl,3b~,5 a~t,9a0h9bfl, 11 afl-perhydrophenanthro-[2,l-b]furan-7-one

C17H2602, M, = 262, triclinic, PI, a = 8.513(2), b = 8.970(2), c = 11.741(3)A, a = 120.51 (5), fl = 93.30(4), y = 68.43(4) ° , V = 708.9,/k 3, Z = 2, D O = 1.213, D e = 1.227 Mg m -a, g(Mo Ka, 2 = 0.7107 ,&) = 0.084 mm -1, F(000) = 288. The structure, solved by direct methods, has been refined to an R value of 5.9% using 1361 intensity measurements. The ring junctions, in sequence from either end of the polycycle, are cis, trans and cis.

X-ray crystal and molecular structure of a hydrated lithium picrate complex of benzo-15-crown-5; An example of non-chelation of the crown

Complexation of alkali and alkaline earth metal ions with crown ethers is well known (1) and chemical and crystallographic studies have been carried out for number of complexes (2,3). The interaction of the metal with the crown ether depends on the nature of the cation and particularly on the basicity of the anion (4) , In this paper we report the crystal and molecular structure of a lithium picrate complex of benzo-15-crown-5, the first x-ray crystallographic study of a lithlum-crown system.

Structure of 2-(2,6-dimethoxyphenyl)-2,5- dimethylbicyclo[3.2.1]octane-6,8-dione

C18H2204, orthorhombic, P212~21, a = 7.343 (4), b = 11.251 (4), c = 19.357 (4)A, Z = 4, Dr, ' = 1.20, D e = 1.254 g cm -3, F(000) = 648, p(MoKa) = 0.94 cm -~. X-ray intensity data were collected on a Nonius CAD-4 diffractometer and the structure was solved by direct methods. Full-matrix least-squares refinement gave R = 0.052 (R w = 0.045) for 1053 observed reflections. The stereochemical configuration at C(2) has been shown to be 2-exo-methyl-2-endo-(2,6-dimethoxyphenyl), i.e. (3) in contrast to the structure (2) assigned earlier based on its ~H NMR data.

Deformation processing of an aluminum alloy containing particles: Studies on AI-5 pct Si alloy 4043

Al-5 wt pct Si alloy is processed by upset forging in the temperature range 300 K to 800 K and in the strain rate range 0.02 to 200 s−1. The hardness and tensile properties of the product have been studied. A “safe” window in the strain rate-temperature field has been identified for processing of this alloy to obtain maximum tensile ductility in the product. For the above strain rate range, the temperature range of processing is 550 K to 700 K for obtaining high ductility in the product. On the basis of microstructure and the ductility of the product, the temperature-strain rate regimes of damage due to cavity formation at particles and wedge cracking have been isolated for this alloy. The tensile fracture features recorded on the product specimens are in conformity with the above damage mechanisms. A high temperature treatment above ≈600 K followed by fairly fast cooling gives solid solution strengthening in the alloy at room temperature.