Effect of cholecalciferol on unsaturated model membranes.

Vitamin D plays an important role in many physiological processes, particularly calcium and phosphorous homeostasis. The biochemistry of vitamin D is also complex, encompassing a range of active molecules that may be either endogenous or dietary in origin. The role of lipids and fats in the production, processing and use of vitamin D is an interesting one, with a relative paucity of model studies into the interactions of vitamin D with lipidic systems such as micelles and vesicles. Here, we have studied the effect of vitamin D3 in simple unsaturated phospholipid systems. We used NMR and FTIR spectroscopy to investigate the effect of increasing vitamin D concentration on the structure and dynamics of the lipid chains and interfacial region. In order to link these model studies with more complex biomimetic environments, we compare results in the presence of buffer and vitamin D binding protein. We have also used DLS to determine that vitamin D3-DOPC vesicles can retain their size distribution for varying amounts of time in different conditions. We find that the acyl chain region of vitamin D3-DOPC membranes are generally disordered, and that the addition of buffer and/or protein alters the properties of the interfacial region.

An algorithm for filtering detector instabilities in search of novel non-exponential decay and in conventional half-life determinations.

Recent reports of Solar modulation of beta-decay have reignited interest in whether or not radioactive half-lives are constants. A numerical approach for filtering instrumental effects on residuals is developed, using correlations with atmospheric conditions recorded while counting 204Tl emissions with a Geiger-Müller counter. Half-life oscillations and detection efficiency oscillations can be separated provided their periods are substantially different. A partial uncertainty budget for the 204Tl half-life shows significant decreases to medium-frequency instabilities correlated with pressure and temperature, which suggests that further development may aid general improvements in half-life determinations.

A bi-prism interferometer for hard X-ray photons.

Micro-fabricated bi-prisms have been used to create an interference pattern from an incident hard X-ray beam, and the intensity of the pattern probed with fluorescence from a 30 nm-thick metal film. Maximum fringe visibility exceeded 0.9 owing to the nano-sized probe and the choice of single-crystal prism material. A full near-field analysis is necessary to describe the fringe field intensities, and the transverse coherence lengths were extracted at APS beamline 8-ID-I. It is also shown that the maximum number of fringes is dependent only on the complex refractive index of the prism material.

Kinoform optics applied to X-ray photon correlation spectroscopy.

Moderate-demagnification higher-order silicon kinoform focusing lenses have been fabricated to facilitate small-angle X-ray photon correlation spectroscopy (XPCS) experiments. The geometric properties of such lenses, their focusing performance and their applicability for XPCS measurements are described. It is concluded that one-dimensional vertical X-ray focusing via silicon kinoform lenses significantly increases the usable coherent flux from third-generation storage-ring light sources for small-angle XPCS experiments.

Diamond kinoform hard X-ray refractive lenses: design, nanofabrication and testing.

Motivated by the anticipated advantageous performance of diamond kinoform refractive lenses for synchrotron X-ray radiation studies, this report focuses on progress in designing, nanofabricating and testing of their focusing performance. The method involves using lift-off and plasma etching to reproduce a planar definition of numerically determined kinoform refractive optics. Tests of the focusing action of a diamond kinoform refractive lens at the APS 8-ID-I beamline demonstrate angular control of the focal spot.

Dual antiglioma action of metformin: cell cycle arrest and mitochondria-dependent apoptosis.

The present study reports for the first time a dual antiglioma effect of the well-known antidiabetic drug metformin. In low-density cultures of the C6 rat glioma cell line, metformin blocked the cell cycle progression in G(0)/G(1) phase without inducing significant cell death. In confluent C6 cultures, on the other hand, metformin caused massive induction of caspase-dependent apoptosis associated with c-Jun N-terminal kinase (JNK) activation, mitochondrial depolarization and oxidative stress. Metformin-triggered apoptosis was completely prevented by agents that block mitochondrial permeability transition (cyclosporin A) and oxygen radical production (N-acetylcisteine), while the inhibitors of JNK activation (SP600125) or glycolysis (sodium fluoride, iodoacetate) provided partial protection. The antiglioma effect of metformin was reduced by compound C, an inhibitor of AMP-activated protein kinase (AMPK), and was mimicked by the AMPK agonist AICAR. Similar effects were observed in the human glioma cell line U251, while rat primary astrocytes were completely resistant to the antiproliferative and proapoptotic action of metformin.

Mechanism of 3-deazaguanine transport in the rat heart.

The aim of this study was to determine the mechanism of transport of 3-deazaguanine in the rat heart. We used single-pass, paired-tracer dilution method on isolated and retrogradely perfused rat hearts. The maximal cellular uptake (Umax) and total cellular uptake (Utot) of 3-deazaguanine were determined under control conditions and under influence of possible modifiers. Both Umax and Utot were significantly reduced in the presence of unlabeled 3-deazaguanine (from 19.57 +/- 2.02% to 8.14 +/- 1.19% and from 16.49 +/- 3.65% to 4.70 +/- 1.96%, n=6, respectively). The presence of pyrimidine nucleoside thymidine caused the reduction of both Umax and Utot (from 20.03 +/- 3.76% to 13.58 +/- 3.16% and from 16.43 +/- 3.58% to 11.94 +/- 3.13%, n=6, respectively). Also, we tested the effect of the absence of sodium ions in perfusion solution (both Umax and Utot, significantly reduced from 17.95 +/- 2.73% to 16.67 +/- 2.16% and from 16.68 +/- 2.97% to 14.81 +/- 3.04%, n=6, respectively) and the effect of dinitrophenol (both Umax and Utot significantly reduced from 19.09 +/- 3.68% to 10.58 +/- 3.14% and from 16.86 +/- 3.84% to 7.10 +/- 3.11%, n=6, respectively). The results of self- and cross-inhibition studies show that the transport of 3-deazaguanine is saturable, energy- and sodium-dependent and that 3-deazaguanine uses endogenous transport systems for thymidine and adenosine for its own transport.

Shear modulus and plasticity of a driven charge density wave.

We have probed the effects of transverse variations in pinning strength on charge-density-wave (CDW) structure in NbSe3 by x-ray micro-beam diffraction. In ribbonlike crystals having a large longitudinal step in thickness, the CDW first depins on the thick side of the step, causing rotations of the CDW wave vector. By measuring these rotations as a function of position and electric field, the corresponding shear strains are determined, allowing the CDW's shear modulus to be estimated. These results demonstrate the usefulness of x-ray microdiffraction as a tool in studying collective dynamics in electronic crystals.

Dynamic nuclear polarization by electrical spin injection in ferromagnet-semiconductor heterostructures.

Electrical spin injection from Fe into AlxGa1-xAs quantum well heterostructures is demonstrated in small (<500 Oe) in-plane magnetic fields. The measurement is sensitive only to the component of the spin that precesses about the internal magnetic field in the semiconductor. This field is much larger than the applied field and depends strongly on the injection current density. Details of the observed hysteresis in the spin injection signal are reproduced in a model that incorporates the magnetocrystalline anisotropy of the epitaxial Fe film, spin relaxation in the semiconductor, and the dynamic polarization of nuclei by the injected spins.

The kinetics of hypoxanthine efflux from the rat brain.

The brain efflux of radiolabelled hypoxanthine in the rat was rapid in the first minute after injection [K(eff)(i)=0.21+/-0.06 min(-1)], which was saturable with a V(max)=13.08+/-0.81 nM min(-1) g(-1), and a high K(m,app) (67.2+/-13.4 microM); the K(i,app) for inosine was 31.5+/-7.6 microM. Capillary depletion analysis indicated that hypoxanthine accumulates in neurons and glia with the time. From cross-inhibition studies with different purines and pyrimidines, it suggests that these molecules could also be important substrates for this carrier.

The characteristics of nucleobase transport and metabolism by the perfused sheep choroid plexus.

The uptake of nucleobases was investigated across the basolateral membrane of the sheep choroid plexus perfused in situ. The maximal uptake (U(max)) for hypoxanthine and adenine, was 35.51+/-1.50% and 30.71+/-0.49% and for guanine, thymine and uracil was 12.00+/-0.53%, 13.07+/-0.48% and 12.30+/-0.55%, respectively with a negligible backflux, except for that of thymine (35.11+/-5.37% of the U(max)). HPLC analysis revealed that the purine nucleobase hypoxanthine and the pyrimidine nucleobase thymine can pass intact through the choroid plexus and enter the cerebrospinal fluid CSF so the lack of backflux for hypoxanthine was not a result of metabolic trapping in the cell. Competition studies revealed that hypoxanthine, adenine and thymine shared the same transport system, while guanine and uracil were transported by a separate mechanism and that nucleosides can partially share the same transporter. HPLC analysis of sheep CSF collected in vivo revealed only two nucleobases were present adenine and hypoxanthine; with an R(CSF/Plasma) 0.19+/-0.02 and 3.43+/-0.20, respectively. Xanthine and urate, the final products of purine catabolism, could not be detected in the CSF even in trace amounts. These results suggest that the activity of xanthine oxidase in the brain of the sheep is very low so the metabolic degradation of purines is carried out only as far as hypoxanthine which then accumulates in the CSF. In conclusion, the presence of saturable transport systems for nucleobases at the basolateral membrane of the choroidal epithelium was demonstrated, which could be important for the distribution of the salvageable nucleobases, adenine and hypoxanthine in the central nervous system.

The kinetics of tiazofurin uptake by the isolated perfused choroid plexus of the sheep.

Tiazofurin (TZF-beta-D-ribofuronosyl thiazole-4-carboxamide, NSC-286193) is a synthetic nucleoside analog with potent antitumor activity. Isolated choroid plexuses (CP) of sheep were perfused in situ and the uptake of [3H]-tiazofurin was determined in relation to the recovery of [14C]-mannitol by means of the paired indicator dilution technique. The maximal uptake of tiazofurin was 8.29 +/- 0.84% and was shown to be both carrier-mediated, sodium-dependent and inhibited by adenosine which suggests that it uses the carrier for endogenous nucleosides. However, the total tiazofurin uptake into the choroid plexus was negligible (0.93 +/- 1.97%) as a result of a high backflux, indicating that tiazofurin is not trapped within the cells of the CP to any significant degree. The kinetics for the uptake into the CP were more favorable than for its passage across the blood-brain barrier with a Km of 7.71 +/- 1.42 microM, a Vmax of 1.30 +/- 0.05 microM/min/g and a negligible constant of a free diffusion (Kd) which suggests that the CP/CSF route may act as an alternative pathway into the brain.