PCR analysis of the wheat varieties and near-isogenic wheat lines with the use of allele-specific primers for the <Emphasis Type="Italic">Gli-1</Emphasis> and <Emphasis Type="Italic">Glu-3</Emphasis> loci

The allelic characteristics of Gli-A1, Gli-B1, Gli-D1 and Glu-A3 loci of 14 bread wheat varieties and 6 near-isogenic wheat lines derived from the Bezosta 1 variety were found by the use of PCR. The conformity between molecular-genetic and storage protein electrophoretic data was revealed: the GliA1.2 allele corresponds to the Gli-A1o and Gli-A1m allelic variants of gliadin blocks; the GliA1.1 PCR allele corresponds to the Gli-A1f, Gli-A1b and Gli-A1c variants of gliadin blocks; the GliB1.1 allele corresponds to the Gli-B1b and Gli-B1d allelic variants; and the GliB1.2 allele corresponds to the Gli-B1e, Gli-B1g and Gli-B1c variants. A new PCR allele with primers for marker GliB1.1 at the Gli-B1 locus in the GLI-B1-12 line (with the gliadin Gli-Blo block), which was generated from crossing of Bezosta 1 and the variety Levent, was detected.     

Mechanism for translocation of fluoroquinolones across lipid membranes

Classical atom-scale molecular dynamics simulations, constrained free energy calculations, and quantum mechanical (QM) calculations are employed to study the diffusive translocation of ciprofloxacin (CPFX) across lipid membranes. CPFX is considered here as a representative of the fluoroquinolone antibiotics class. Neutral and zwitterionic CPFX coexist at physiological pH, with the latter being predominant. Simulations reveal that only the neutral form permeates the bilayer, and it does so through a novel mechanism that involves dissolution of concerted stacks of zwitterionic ciprofloxacins. Subsequent QM analysis of the observed molecular stacking shows the important role of partial charge neutralization in the stacks, highlighting how the zwitterionic form of the drug is neutralized for translocation. The findings propose a translocation mechanism in which zwitterionic CPFX molecules approach the membrane in stacks, but they diffuse through the membrane as neutral CPFX monomers due to intermolecular transfer of protons favored by partial solvation loss. The mechanism is expected to be of importance in the permeation and translocation of a variety of ampholitic drugs with stacking tendencies.     

A 14-3-3γ dimer-based scaffold bridges CtBP1-S/BARS to PI(4)KIIIβ to regulate post-Golgi carrier formation

Large pleiomorphic carriers leave the Golgi complex for the plasma membrane by en bloc extrusion of specialized tubular domains, which then undergo fission. Several components of the underlying molecular machinery have been identified, including those involved in the budding/initiation of tubular carrier precursors (for example, the phosphoinositide kinase PI(4)KIIIβ, the GTPase ARF, and FAPP2), and in the fission of these precursors (for example, PKD, CtBP1-S/BARS). However, how these proteins interact to bring about carrier formation is poorly understood. Here, we describe a protein complex that mediates carrier formation and contains budding and fission molecules, as well as other molecules, such as the adaptor protein 14-3-3γ. Specifically, we show that 14-3-3γ dimers bridge CtBP1-S/BARS with PI(4)KIIIβ, and that the resulting complex is stabilized by phosphorylation by PKD and PAK. Disrupting the association of these proteins inhibits the fission of elongating carrier precursors, indicating that this complex couples the carrier budding and fission processes.     

The role of vitamin E in the biosynthesis of ubiquinone (Q) and ubichromenol (QC) in rat liver

Some peculiarities of ubiquinone (Q) biosynthesis in the livers of vitamin E-deficient rats induced in vitro by alpha-tocopherol and S-adenosylmethionine (SAM) were investigated. When [C3H3]SAM was used as a precursor, alpha-tocopherol added to the sample induced a marked elevation of the content of Q concomitantly with a drastic increase (2.82-fold) of specific radioactivity of Q in the liver as compared to control. Under identical conditions with 2-14C sodium acetate as precursor, exogenous alpha-tocopherol increased the content (by 31.4%) and specific radioactivity (by 65.2%) of Q in experimental samples, whereas the corresponding parameters of ubichromenol (QC) were essentially unchanged. SAM added to the incubation medium caused a marked increase in the content and specific radioactivity of both Q and QC as compared to control. Similar, and even more conspicuous changes were observed after combined administration of alpha-tocopherol and SAM. The role of vitamin E in the activation of methylation reactions at terminal steps of Q and QC biosynthesis in rat liver is discussed.     

The endometrial glands at various phases of the menstrual cycle (morphometric research)

In diagnostic curettages of the uterine cavity wall in 14 women 878 glands have been investigated. The following parameters have been estimated: perimeters of the gland section contours and their lumens, areas of the gland sections, their lumens and epithelial rings, coefficients of the gland forms and their lumens, distance between glands. Peculiarities connected with physiological differences of the uterine cavity mucose membrane are revealed at various phases of the ovarian menstrual cycle     

RGD mimetics containing phthalimidine fragment as novel ligands of fibrinogen receptor

The novel RGD mimetics with phthalimidine central fragment were synthesized with the use of 4-piperidine-4-yl-butyric, 4-piperidine-4-yl-benzoic, 4-piperazine-4-yl-benzoic and 1,2,3,4-tetrahydroisoquinoline-7-carboxylic acids as surrogates of Arg motif. The synthesized compounds potently inhibited platelet aggregation in vitro and blocked FITC-Fg binding to α(IIb)β(3) integrin in a suspension of washed human platelets. The key α(IIb)β(3) protein-ligand interactions were determined in docking experiments.     

Docking studies on novel analogues of 8 methoxy fluoroquinolones against GyrA mutants of Mycobacterium tuberculosis

Background

Modeling of transmembrane domains (TMDs) requires correct prediction of interfacial residues for in-silico modeling and membrane insertion studies. This implies the defining of a target sequence long enough to contain interfacial residues. However, too long sequences induce artifactual polymorphism: within tested modeling methods, the longer the target sequence, the more variable the secondary structure, as though the procedure were stopped before the end of the calculation (which may in fact be unreachable). Moreover, delimitation of these TMDs can produce variable results with sequence based two-dimensional prediction methods, especially for sequences showing polymorphism. To solve this problem, we developed a new modeling procedure using the PepLook method. We scanned the sequences by modeling peptides from the target sequence with a window of 19 residues.

Results

Using sequences whose NMR-structures are already known (GpA, EphA1 and Erb2-HER2), we first determined that the hydrophobic to hydrophilic accessible surface area ratio (ASAr) was the best criterion for delimiting the TMD sequence. The length of the helical structure and the Impala method further supported the determination of the TMD limits. This method was applied to the IL-2Rβ and IL-2Rγ TMD sequences of Homo sapiens, Rattus norvegicus, Mus musculus and Bos taurus.

Conclusions

We succeeded in reducing the variation in the TMD limits to only 2 residues and in gaining structural information.