Novel dinucleoside 5',5'-triphosphate cap analogues. Synthesis and affinity for murine translation factor eIF4E

Chemical synthesis of a series of novel dinucleoside cap analogues, m7GpppN, where N is formycin A, 3'-O-methylguanosine, 9-beta-D-arabinofuranosyladenine, and isoguanosine, has been performed using our new methodology. The key reactions of pyrophosphate bonds formation were achieved in anhydrous dimethylformamide solutions employing the catalytic properties of zinc salts. Structures of the new cap analogues were confirmed by 1H NMR and 31p NMR spectra. The binding affinity of the new cap analogues for murine eIF4E(28-217) were determined spectroscopically showing the highest association constant for the analogue that contains formycin A.     

Folding thermodynamics and kinetics of YNMG RNA hairpins: specific incorporation of 8-bromoguanosine leads to stabilization by enhancement of the folding rate

Modified nucleotides allow fundamental energetic and kinetic properties of nucleic acids to be probed. Here, we demonstrate that an RNA hairpin containing the nucleotide analogue 8-bromoguanosine (8BrG or G), gcUUCGgc, has enhanced stability relative to the unmodified hairpin, with DeltaDeltaG(37)(degrees)= -0.69 +/- 0.15 kcal mol(-1) and DeltaT(M) = +6.8 +/- 1.4 degrees C. NMR spectroscopic data suggest that the enhanced stability of gcUUCGgc does not arise from the native state; laser temperature-jump experiments support this notion, as gcUUCGgc and gcUUCGgc have similar unfolding rate constants, but the folding rate constant of gcUUCGgc is 4.1-fold faster at 37.5 degrees C and 2.8-fold faster under isoenergetic conditions. On the basis of these findings, we propose that 8BrG reduces the conformational entropy of the denatured state, resulting in an accelerated conformational search for the native state and enhanced stability.     

Compensating bends in a 16-base-pair DNA oligomer containing a T(3)A(3) segment: A NMR study of global DNA curvature

In-phase ligated DNA containing T(n)A(n) segments fail to exhibit the retarded polyacrylamide gel electrophoresis (PAGE) migration observed for in-phase ligated A(n)T(n) segments, a behavior thought to be correlated with macroscopic DNA curvature. The lack of macroscopic curvature in ligated T(n)A(n) segments is thought to be due to cancellation of bending in regions flanking the TpA steps. To address this issue, solution-state NMR, including residual dipolar coupling (RDC) restraints, was used to determine a high-resolution structure of [d(CGAGGTTTAAACCTCG)2], a DNA oligomer containing a T3A3 tract. The overall magnitude and direction of bending, including the regions flanking the central TpA step, was measured using a radius of curvature, Rc, analysis. The Rc for the overall molecule indicated a small magnitude of global bending (Rc = 138 +/- 23 nm) towards the major groove, whereas the Rc for the two halves (72 +/- 33 nm and 69 +/- 14 nm) indicated greater localized bending into the minor groove. The direction of bending in the regions flanking the TpA step is in partial opposition (109 degrees), contributing to cancellation of bending. The cancellation of bending did not correlate with a pattern of roll values at the TpA step, or at the 5' and 3' junctions, of the T3A3 segment, suggesting a simple junction/roll model is insufficient to predict cancellation of DNA bending in all T(n)A(n) junction sequence contexts. Importantly, Rc analysis of structures refined without RDC restraints lacked the precision and accuracy needed to reliably measure bending.     

Assessment of various diagnostic methods of ureaplasma respiratory tract infections in newborns

We compared three methods used microbial culturing for detection of ureaplasmas in endotracheal aspirate from 500 prematurely born neonates with respiratory disturbances: BioMerieux test, PCR and microbial culturing. Ureaplasmas were detected in respiratory tracts of 79 (16%) newborns. Correlation of the results of culture with those obtained with the BioMerieux kit, culture with PCR and BioMerieux kit with PCR was 97%, 89% and 90%, respectively. Sensitivity and specificity of PCR in comparison with culture was 86% and 98%, respectively, and of the BioMerieux kit 96% and 98%. PCR can be recommended in rapid diagnostics of respiratory infections in newborns suffering from respiratory disorders. It allows the detection of ureaplasmas in case of parallel infections and identification of their species.     

Cieneguiticeras, a new genus of Tithonian oppeliids (Ammonoidea, Late Jurassic)

The new genus Cieneguiticeras, assigned to the family Oppeliidae, is described on the basis of Andean lower-middle Tithonian ammonites from Arroyo Cieneguita, west-central part of the Neuquén-Mendoza Basin, Argentina. The macroconchs are closely homoeomorphic with Neochetoceras Spath and the microconchs have a ‘glochiceratid’-type morphology. The stratigraphic range of Cieneguiticeras nov. gen. includes the lower and middle Tithonian by means of a succession of two or three species which are interpreted as members of a phyletic lineage. Ammonites from the Tithonian of Cuba, Mexico and France are more or less confidently included in this new genus.     

Structural and spectroscopic characterisation of the holmium double-decker partially oxidised and bi-radical phthalocyanine complexes with iodine

Two crystals of holmium(III) double-decker iodine doped phthalocyanines, HoPc₂I_5/3 (I) and HoPc₂I (II), were grown directly in the reaction of holmium chips with 1,2-dicyanobenzene under versatile quantity of iodine at 180-160 °C. The complex I crystallises in the P4/mcc space group of tetragonal system, while the complex II crystallises in the P2/c space group of monoclinic system. The space group of P4/mcc and z = 1 requires that the Ho(III) atom is statistically disordered in the HoPc₂I_5/3 structure. The iodine atoms form linear symmetrical triiodide ions in I, while the I⁻ ions in II. Assignment of iodine species as in the HoPc₂I_5/3 and I⁻ in HoPc₂I complexes point to the +5/9 and +1 oxidation state of the HoPc₂ unit in these complexes. Thus one Pc macrocycle of the double-decker HoPc₂ units has a non-integer oxidation state of −1.222 in I, while both Pc-rings are one-electron oxidised radical Pc⁻ in II. Magnetic susceptibilities of HoPc₂I_5/3 and HoPcI at room temperature are 4.56 × 10⁻² and 5.12 × 10⁻² emu/mol and the calculated magnetic moments are 10.46 and 11.08 μ_B, respectively. UV-Vis spectroscopic measurement of I and II in benzene solution were carried out and discussed.     

Psychological view of thyroid cancer and in hyperthyroid Graves' patients - comparative exploration

INTRODUCTION: The aim of study was the evaluation of association between psychometric factors in patients with thyroid cancer and in Graves' hyperthyroid patients. MATERIAL AND METHODS: We examined 50 patients with differentiated thyroid cancer, 42 females (84.75%), 8 males aged from 32 to 64 yr. (x +/- SD: 43 +/- 8.17 yr.) and 50 hyperthyroid patients, 45 (90%) females and 5 (10%) males corresponding aged. We used the following methods: the EAS Temperament Survey (EASD) in addition for adults, EPQ Eysenck Personality Questionnaire, Polish Abbreviated Form of the MMPI (DKO-74) and Beck's Depression Scale. We compared the scores of thyroid cancer patients and hyperthyroid patients. RESULTS: We have interpreted obtained results in both groups as similarly in: mild sense depression, high emotional control, high social dependence, ambiverssion, mild level of emotional mental balance. CONCLUSION: Results of our explorations appears great psychological similarity between differentiated thyroid patients and hyperthyroid patients.     

Bacteriophage translocation

The occurrence of phages in the human body, especially in the gastrointestinal tract, raises the question of their potential role in the physiology and pathology of this system. Especially important is the issue of whether phages can pass the intestinal wall and migrate to lymph, peripheral blood, and internal organs and, if so, the effects such a phenomenon could have (such passage by bacteria, known as bacterial translocation, has been shown to cause various disturbances in humans, from immune defects to sepsis). Available data from the literature support the assumption that phage translocation can take place and may have some immunomodulatory effects. In addition, phages of the gut may play a protective role by inhibiting local immune reactions to antigens derived from gut flora.     

Loop formation by the transgeneWAP:6xHishGH in transgenic rabbit fibroblasts, revealed by fluorescencein situ hybridization to nuclear halos

Using fluorescence in situ hybridization (FISH) to somatic nuclear halos from transgenic rabbits WAP:6xHishGH, we present evidence for stability of transgenesis at the chromatin level. FISH performed on fibroblasts from a homozygous individual showed 2 independent loops from both chromosomes of pair 7. On a heterozygous individual, FISH detected a single loop. According to the concept of chromatin loops and their influence on gene expression, this shows that the human growth hormone transgene, which was actively expressed in mammary gland under the influence of the tissue-specific promoter, was inactive in examined skin fibroblasts.