In vitro release of nonoxynol-9 from silicone matrix intravaginal rings

The controlled-release characteristics of matrix silicone intravaginal rings loaded with between 100 and 971 mg of nonoxynol-9 have been investigated with a view to developing a ring that may offer a new female-controlled method for the prevention of transmission of sexually transmitted diseases, particularly HIV. Intravaginal rings containing 253, 487 and 971 mg of nonoxynol-9 provided a daily release of 2 mg or more over the 8-day release period, the minimal amount of nonoxynol-9 considered to provide an effective vaginal concentration for the prevention of HIV. Furthermore, the maximum daily release of N9 was about 6 mg, an amount significantly smaller than that observed for other nonoxynol-9 products whose large daily doses may in fact increase the occurrence of HIV by causing epithelial damage to the vaginal tissue. The release mechanism of the liquid nonoxynol-9 from the intravaginal rings has also been investigated and compared to models describing the release of solid drugs from the rings. It has been demonstrated through release studies and surface microscopy that a drug depletion zone is not established in such liquid-loaded intravaginal ring systems, with implications for the release kinetics. (C) 2003 Elsevier B.V. All rights reserved.

Enhanced Deuterium Fractionation in Dense Interstellar Cores Resulting from Multiply Deuterated H3+

In this Letter, we present the first results from updated models of interstellar deuterium chemistry that now include all possible deuterated isotopomers of H3+. We find that in regions of high density and heavy depletion, such as prestellar cores, the inclusion of HD2+ and D3+ enhances the fractionation of ionic and neutral species significantly. Our models are the first to predict the very high atomic D/H ratios (>=0.3) necessary for grain-surface chemistry models to reproduce the high formaldehyde and methanol fractionation seen in star-forming regions.

Ascorbic acid in nasal and tracheobronchial airway lining fluids

Ascorbic acid (AA) is thought to be an important antioxidant in the respiratory tract, whose regulation is yet to be fully characterized. We investigated whether AA in respiratory tract lining fluids (RTLFs) can be augmented by oral supplementation with AA. Plasma, nasal lavage fluids (NLFs), induced sputum (IS), and saliva were analyzed for AA immediately before and 2 h after ingestion of 2 g of AA in 13 healthy subjects. Concentrations of AA (median and range) were 52.5 (16.0-88.5), 2.4 (0.18-4.66), 2.4 (0.18-6.00), and 0.55 (0.18-18.90) micromol/l, respectively. Two hours after ingestion of AA, plasma AA increased 2-fold (p = .004), NLF AA increased 3-fold (p = .039), but IS and saliva AA did not increase. As AA concentrations in saliva and tracheobronchial secretions were low compared with other common extracellular components (such as urate), we evaluated the fate of AA in these fluids. Addition of AA to freshly obtained saliva or IS resulted in rapid depletion, which could be largely prevented or reversed by sodium azide or dithiothreitol. These findings suggest that oxidant-producing systems in saliva and airway secretions, such as heme peroxidases and other oxidizing substances, rapidly consume AA. Whereas oral supplementation resulted in detectable increases of AA in NLFs, its levels in tracheobronchial lining fluid, as measured by IS, were unaffected and remained relatively low, suggesting that AA may play a less significant antioxidant role in this compartment as compared with most other extracellular compartments.

Carbachol triggers RyR-dependent Ca(2+) release via activation of IP(3) receptors in isolated rat gastric myocytes.

Possible interactions between different intracellular Ca(2+) release channels were studied in isolated rat gastric myocytes using agonist-evoked Ca(2+) signals. Spontaneous, local Ca(2+) transients were observed in fluo-4-loaded cells with linescan confocal imaging. These were blocked by ryanodine (100 microM) but not by the inositol 1,4,5-trisphosphate receptor (IP(3)R) blocker, 2-aminoethoxydiphenyl borate (100 microM), identifying them as Ca(2+) sparks. Caffeine (10 mM) and carbachol (10 microM) initiated Ca(2+) release at sites which co-localized with each other and with any Ca(2+) spark sites. In fura-2-loaded cells extracellular 2-aminoethoxydiphenyl borate and intracellular heparin (5 mg ml(-1)) both inhibited the global cytoplasmic [Ca(2+)] transient evoked by carbachol, confirming that it was IP(3)R-dependent. 2-Aminoethoxydiphenyl borate and heparin also increased the response to caffeine. This probably reflected an increased Ca(2+) store content since 2-aminoethoxydiphenyl borate more than doubled the amplitude of transients evoked by ionomycin. Ryanodine completely abolished carbachol and caffeine responses but only reduced ionomycin transients by 30 %, suggesting that blockade of carbachol transients by ryanodine was not simply due to store depletion. Double labelling of IP(3)Rs and RyRs demonstrated extensive overlap in their distribution. These results suggest that carbachol stimulates Ca(2+) release through co-operation between IP(3)Rs and RyRs, and implicate IP(3)Rs in the regulation of Ca(2+) store content.

Restoration of glutathione levels in vascular smooth muscle cells exposed to high glucose conditions

Hyperglycaemia-induced oxidative stress may play a key role in the pathogenesis of diabetic vascular disease. The purpose of the present study was to determine the effects of glucose on levels of glutathione (a major intracellular antioxidant), the expression of gamma-glutamylcysteine synthetase (the rate-limiting enzyme in glutathione de novo synthesis) and DNA damage in human vascular smooth muscle cells in vitro. High glucose conditions and buthionine sulphoximine, an inhibitor of gamma-glutamylcysteine synthetase, reduced intracellular glutathione levels in vascular smooth muscle cells. This reduction was accompanied by a decrease in the mRNA expression of both subunits of gamma-glutamylcysteine synthetase as well as an increase in DNA damage. In high glucose conditions incubation of the vascular smooth muscle cells with alpha-lipoic acid and L-cystine restored glutathione levels. We suggest that the decrease in GSH levels seen in high glucose conditions is mediated by the availability of cysteine (rate-limiting substrate in de novo glutathione synthesis) and the gene expression of the gamma- glutamylcysteine synthetase enzyme. Glutathione depletion is associated with an increase in DNA damage, which can be reduced when glutathione levels are restored.

Down-regulation of Rac Activity during beta 2 Integrin-mediated Adhesion of Human Neutrophils

In human neutrophils, beta2 integrin engagement mediated a decrease in GTP-bound Rac1 and Rac2. Pretreatment of neutrophils with LY294002 or PP1 (inhibiting phosphatidylinositol 3-kinase (PI 3-kinase) and Src kinases, respectively) partly reversed the beta2 integrin-induced down-regulation of Rac activities. In contrast, beta2 integrins induced stimulation of Cdc42 that was independent of Src family members. The PI 3-kinase dependency of beta2 integrin-mediated decrease in GTP-bound Rac could be explained by an enhanced Rac-GAP activity, since this activity was blocked by LY204002, whereas PP1 only had a minor effect. The fact that only Rac1 but not Rac2 (the dominating Rac) redistributed to the detergent-insoluble fraction and that it was independent of GTP loading excludes the possibility that down-regulation of Rac activities was due to depletion of GTP-bound Rac from the detergent-soluble fraction. The beta2 integrin-triggered relocalization of Rac1 to the cytoskeleton was enabled by a PI 3-kinase-induced dissociation of Rac1 from LyGDI. The dissociations of Rac1 and Rac2 from LyGDI also explained the PI 3-kinase-dependent translocations of Rac GTPases to the plasma membrane. However, these accumulations of Rac in the membrane, as well as that of p47phox and p67phox, were also regulated by Src tyrosine kinases. Inasmuch as Rac GTPases are part of the NADPH oxidase and the respiratory burst is elicited in neutrophils adherent by beta2 integrins, our results indicate that activation of the NADPH oxidase does not depend on the levels of Rac-GTP but instead requires a beta2 integrin-induced targeting of the Rac GTPases as well as p47phox and p67phox to the plasma membrane.

Laboratory trials on the effects of different diets on growth and survival of the common whelk, Buccinum undatum L. 1758, as a candidate species for aquaculture.

Newly hatched juvenile Buccinum undatum can be reared under laboratory conditions. Good was growth is achieved when juveniles were fed on combined diets (blue mussel, cod, and fish pellets). Juveniles reached shell heights of 33.0 ± 4.2 mm, 26.9 ± 3.8 ± mm, 23.2 ± 2.2 mm, and 20.1 ± 1.6 mm, after 14 months of fedding on a combined diet, blue mussel, cod, and fish pellets, respectively under ambient sea temperature and salinity. After 14 months juveniles fed blue mussel had the highest survival rates (67%) followed by those fed a combination of all other experimental diets (61%), cod waste (53%) and fish-feed pellets (46%). High mortalities were recorded in most treatments during the summer months between June and September. This species appears to have an aquaculture potential, as juveniles readily feed on artificial diets at an early age, show high survival rates and could potentially reach market size in 2 years or less. The major constraint in realising this potential at present, is the relatively low value of the species; if market values increased as a result of serious depletion of natural populations, hatchery production of juveniles for intensive aquaculture or restocking could become economically viable.

Environmental Isotopes as Indicators for Ground Water Recharge to Fractured Granite

To assess the contribution of accumulated winter precipitation and glacial meltwater to the recharge of deep ground water flow systems in fracture crystalline rocks, measurements of environmental isotope ratios, hydrochemical composition, and in situ parameters of ground water were performed in a deep tunnel. The measurements demonstrate the significance of these ground water recharge components for deep ground water flow systems in fractured granites of a high alpine catchment in the Central Alps, Switzerland. Hydrochemical and in situ parameters, as well as d18O in ground water samples collected in the tunnel, show only small temporal variations. The precipitation record of d18O shows seasonal variations of ~14‰ and a decrease of 0.23‰ ± 0.03‰ per 100 m elevation gain. d2H and d18O in precipitation are well correlated and plot close to the meteoric water line, as well as d2H and d18O in ground water samples, reflecting the meteoric origin of the latter. The depletion of 18O in ground water compared to 18O content in precipitation during the ground water recharge period indicates significant contributions from accumulated depleted winter precipitation to ground water recharge. The hydrochemical composition of the encountered ground water, Na-Ca-HCO3-SO4(-F), reflects an evolution of the ground water along the flowpath through the granite body. Observed tritium concentrations in ground water range from 2.6 to 16.6 TU, with the lowest values associated with a local negative temperature anomaly and anomalous depleted 18O in ground water. This demonstrates the effect of local ground water recharge from meltwater of submodern glacial ice. Such localized recharge from glaciated areas occurs along preferential flowpaths within the granite body that are mainly controlled by observed hydraulic active shear fractures and cataclastic faults.

Early-type stars observed in the ESO UVES Paranal Observatory Project - I. Interstellar NaIUV, TiII and CaIIKobservations

We present an analysis of interstellar NaI (lambda(air) = 3302.37 and 3302.98 angstrom), TiII (lambda(air) = 3383.76 angstrom) and CaII K (lambda(air) = 3933.66 angstrom) absorption features for 74 sightlines towards O- and B-type stars in the Galactic disc. The data were obtained from the Ultraviolet and Visual Echelle Spectrograph Paranal Observatory Project, at a spectral resolution of 3.75 km s(-1) and with mean signal-to-noise ratios per pixel of 260, 300 and 430 for the NaI, TiII and CaII observations, respectively. Interstellar features were detected in all but one of the TiII sightlines and all of the CaII sightlines. The dependence of the column density of these three species with distance, height relative to the Galactic plane, HI column density, reddening and depletion relative to the solar abundance has been investigated. We also examine the accuracy of using the NaI column density as an indicator of that for HI. In general, we find similar strong correlations for both Ti and Ca, and weaker correlations for Na. Our results confirm the general belief that Ti and Ca occur in the same regions of the interstellar medium ( ISM) and also that the TiII/CaII ratio is constant over all parameters. We hence conclude that the absorption properties of Ti and Ca are essentially constant under the general ISM conditions of the Galactic disc.

Ferroelectricity and isotope effects in hydrogen-bonded KDP crystals

Based on an accurate first principles description of the energetics in H-bonded potassium-dihydrogen-phosphate crystals, we conduct a first study of nuclear quantum effects and of the changes brought about by deuteration. Tunneling is allowed only for clusters involving correlated protons and heavy ion displacements, the main effect of deuteration being a depletion of the proton probability density at the O-H-O bridge center, which in turn weakens its proton-mediated covalent bonding. The ensuing lattice expansion couples self-consistently with the proton off-centering, thus explaining both the giant isotope effect and its close connection with geometrical effects.

Molecular effects in the ionization of N-2, O-2, and F-2 by intense laser fields

In this paper we study the response in time of N2, O2, and F2 to laser pulses having a wavelength of 390 nm. We find single-ionization suppression in O2 and its absence in F2, in accordance with experimental results at lambda= 800 nm. Within our framework of time-dependent density functional theory we are able to explain deviations from the predictions of intense-field many-body S-matrix theory (IMST). We confirm the connection of ionization suppression with destructive interference of outgoing electron waves from the ionized electron orbital. However, the prediction of ionization suppression, justified within the IMST approach through the symmetry of the highest occupied molecular orbital (HOMO), is not reliable since it turns out that—e.g., in the case of F2—the electronic response to the laser pulse is rather complicated and does not lead to dominant depletion of the HOMO. Therefore, the symmetry of the HOMO is not sufficient to predict ionization suppression. However, at least for F2, the symmetry of the dominantly ionized orbital is consistent with the nonsuppression of ionization.

Identification of a novel population of Langerin+ dendritic cells.

Langerhans cells (LCs) are antigen-presenting cells that reside in the epidermis of the skin and traffic to lymph nodes (LNs). The general role of these cells in skin immune responses is not clear because distinct models of LC depletion resulted in opposite conclusions about their role in contact hypersensitivity (CHS) responses. While comparing these models, we discovered a novel population of LCs that resides in the dermis and does not represent migrating epidermal LCs, as previously thought. Unlike epidermal LCs, dermal Langerin(+) dendritic cells (DCs) were radiosensitive and displayed a distinct cell surface phenotype. Dermal Langerin(+) DCs migrate from the skin to the LNs after inflammation and in the steady state, and represent the majority of Langerin(+) DCs in skin draining LNs. Both epidermal and dermal Langerin(+) DCs were depleted by treatment with diphtheria toxin in Lang-DTREGFP knock-in mice. In contrast, transgenic hLang-DTA mice lack epidermal LCs, but have normal numbers of dermal Langerin(+) DCs. CHS responses were abrogated upon depletion of both epidermal and dermal LCs, but were unaffected in the absence of only epidermal LCs. This suggests that dermal LCs can mediate CHS and provides an explanation for previous differences observed in the two-model systems.

Tumor immunotherapy by epicutaneous immunization requires langerhans cells.

A role for Langerhans cells (LC) in the induction of immune responses in the skin has yet to be conclusively demonstrated. We used skin immunization with OVA protein to induce immune responses against OVA-expressing melanoma cells. Mice injected with OVA-specific CD8(+) T cells and immunized with OVA onto barrier-disrupted skin had increased numbers of CD8(+) T cells in the blood that produced IFN-gamma and killed target cells. These mice generated accelerated cytotoxic responses after secondary immunization with OVA. Prophylactic or therapeutic immunization with OVA onto barrier-disrupted skin inhibited the growth of B16.OVA tumors. LC played a critical role in the immunization process because depletion of LC at the time of skin immunization dramatically reduced the tumor-protective effect. The topically applied Ag was presented by skin-derived LC in draining lymph nodes to CD8(+) T cells. Thus, targeting of tumor Ags to LC in vivo is an effective strategy for tumor immunotherapy.

Plasma dynamics in an inductively coupled magnetic neutral loop discharge

Magnetic neutral loop discharges (NLDs) can be operated at significantly lower pressures than conventional radio-frequency (rf) inductively coupled plasmas (ICPs). These low pressure conditions are favourable for technological applications, in particular anisotropic etching. An ICP–NLD has been designed providing excellent diagnostics access for detailed investigations of fundamental mechanisms. Spatially resolved Langmuir probe measurements have been performed in the plasma production region (NL region) as well as in the remote application region downstream from the NL region. Depending on the NL gradient two different operation modes have been observed exhibiting different opportunities for control of plasma uniformity. The efficient operation at comparatively low pressures results in ionization degrees exceeding 1%. In this regime neutral dynamics has to be considered and can influence neutral gas and process uniformity. Neutral gas depletion through elevated gas temperatures and high ionization rates have been quantified. At pressures above 0.1 Pa, gas heating is the dominant depletion mechanism. At lower pressures neutral gas is predominantly depleted through high ionization rates and rapid transport of ions by ambipolar diffusion along the magnetic field lines. Non-uniform profiles of the ionization rate can, therefore, result in localized neutral gas depletion and non-uniform processing. We have also investigated the electron dynamics within the radio-frequency cycle using phase resolved optical emission spectroscopy and Thomson scattering. In these measurements electron drift phenomena along the NL torus have been identified.