Endocytosis in filter-grown Madin-Darby canine kidney cells

The nitrogen-containing bisphosphonates used for management of the patients with osteoporosis were reported to influence the function of renal tubular cells. However, how nitrogen-containing bisphosphates exert any effects on ion currents remains controversial. The effects of ibandronate (Iban), a nitrogen-containing bisphosphonate, on ionic channels, including two types of Ca(2+)-activated K(+) (KCa) channels, namely, large-conductance KCa (BKCa) and intermediate-conductance KCa (IKCa) channels, were investigated in Madin-Darby canine kidney (MDCK) cells. In whole-cell current recordings, Iban suppressed the amplitude of voltage-gated K(+) current elicited by long ramp pulse. Addition of Iban caused a reduction of BKCa channels accompanied by a right shift in the activation curve of BKCa channels, despite no change in single-channel conductance. Ca(2+) sensitivity of these channels was modified in the presence of this compound; however, the magnitude of Iban-mediated decrease in BKCa-channel activity under membrane stretch with different negative pressure remained unchanged. Iban suppressed the probability of BKCa-channel openings linked primarily to a shortening in the slow component of mean open time in these channels. The dissociation constant needed for Iban-mediated suppression of mean open time in MDCK cells was 12.2\ua0μM. Additionally, cell exposure to Iban suppressed the activity of IKCa channels, and DC-EBIO or 9-phenanthrol effectively reversed its suppression. Under current-clamp configuration, Iban depolarized the cells and DC-EBIO or PF573228 reversed its depolarizing effect. Taken together, the inhibitory action of Iban on KCa-channel activity may contribute to the underlying mechanism of pharmacological or toxicological actions of Iban and its structurally similar bisphosphonates on renal tubular cells occurring in vivo

Co-infection of the four major Plasmodium species: effects on densities and gametocyte carriage

BACKGROUND: Co-infection of the four major species of human malaria parasite Plasmodium falciparum (Pf), P. vivax (Pv), P. malariae (Pm), and P. ovale sp. (Po) is regularly observed, but there is limited understanding of between-species interactions. In particular, little is known about the effects of multiple Plasmodium species co-infections on gametocyte production. METHODS: We developed molecular assays for detecting asexual and gametocyte stages of Pf, Pv, Pm, and Po. This is the first description of molecular diagnostics for Pm and Po gametocytes. These assays were implemented in a unique epidemiological setting in Papua New Guinea with sympatric transmission of all four Plasmodium species permitting a comprehensive investigation of species interactions. FINDINGS: The observed frequency of Pf-Pv co-infection for asexual parasites (14.7%) was higher than expected from individual prevalence rates (23.8%Pf x 47.4%Pv = 11.3%). The observed frequency of co-infection with Pf and Pv gametocytes (4.6%) was higher than expected from individual prevalence rates (13.1%Pf x 28.2%Pv = 3.7%). The excess risk of co-infection was 1.38 (95% confidence interval (CI): 1.09, 1.67) for all parasites and 1.37 (95% CI: 0.95, 1.79) for gametocytes. This excess co-infection risk was partially attributable to malaria infections clustering in some villages. Pf-Pv-Pm triple infections were four times more frequent than expected by chance alone, which could not be fully explained by infections clustering in highly exposed individuals. The effect of co-infection on parasite density was analyzed by systematic comparison of all pairwise interactions. This revealed a significant 6.57-fold increase of Pm density when co-infected with Pf. Pm gametocytemia also increased with Pf co-infection. CONCLUSIONS: Heterogeneity in exposure to mosquitoes is a key epidemiological driver of Plasmodium co-infection. Among the four co-circulating parasites, Pm benefitted most from co-infection with other species. Beyond this, no general prevailing pattern of suppression or facilitation was identified in pairwise analysis of gametocytemia and parasitemia of the four species. TRIAL REGISTRATION: This trial is registered with ClinicalTrials.gov, Trial ID: NCT02143934

Late Endosomal Cholesterol Accumulation Leads to Impaired Intra-Endosomal Trafficking

Background Pathological accumulation of cholesterol in late endosomes is observed in lysosomal storage diseases such as Niemann-Pick type C. We here analyzed the effects of cholesterol accumulation in NPC cells, or as phenocopied by the drug U18666A, on late endosomes membrane organization and dynamics. Methodology/Principal Findings Cholesterol accumulation did not lead to an increase in the raft to non-raft membrane ratio as anticipated. Strikingly, we observed a 2–3 fold increase in the size of the compartment. Most importantly, properties and dynamics of late endosomal intralumenal vesicles were altered as revealed by reduced late endosomal vacuolation induced by the mutant pore-forming toxin ASSP, reduced intoxication by the anthrax lethal toxin and inhibition of infection by the Vesicular Stomatitis Virus. Conclusions/Significance These results suggest that back fusion of intralumenal vesicles with the limiting membrane of late endosomes is dramatically perturbed upon cholesterol accumulation

Ginzburg-Landau-Gor'kov Theory of Magnetic oscillations in a type-II 2-dimensional Superconductor

We investigate de Haas-van Alphen (dHvA) oscillations in the mixed state of a type-II two-dimensional superconductor within a self-consistent Gor'kov perturbation scheme. Assuming that the order parameter forms a vortex lattice we can calculate the expansion coefficients exactly to any order. We have tested the results of the perturbation theory to fourth and eight order against an exact numerical solution of the corresponding Bogoliubov-de Gennes equations. The perturbation theory is found to describe the onset of superconductivity well close to the transition point $H_{c2}$. Contrary to earlier calculations by other authors we do not find that the perturbative scheme predicts any maximum of the dHvA-oscillations below $H_{c2}$. Instead we obtain a substantial damping of the magnetic oscillations in the mixed state as compared to the normal state. We have examined the effect of an oscillatory chemical potential due to particle conservation and the effect of a finite Zeeman splitting. Furthermore we have investigated the recently debated issue of a possibility of a sign change of the fundamental harmonic of the magnetic oscillations. Our theory is compared with experiment and we have found good agreement.Comment: 39 pages, 8 figures. This is a replacement of supr-con/9608004. Several sections changed or added, including a section on the effect of spin and the effect of a conserved number of particles. To be published in Phys. Rev.

SNX12 Role in Endosome Membrane Transport

In this paper, we investigated the role of sorting nexin 12 (SNX12) in the endocytic pathway. SNX12 is a member of the PX domain-containing sorting nexin family and shares high homology with SNX3, which plays a central role in the formation of intralumenal vesicles within multivesicular endosomes. We found that SNX12 is expressed at very low levels compared to SNX3. SNX12 is primarily associated with early endosomes and this endosomal localization depends on the binding to 3-phosphoinositides. We find that overexpression of SNX12 prevents the detachment (or maturation) of multivesicular endosomes from early endosomes. This in turn inhibits the degradative pathway from early to late endosomes/lysosomes, much like SNX3 overexpression, without affecting endocytosis, recycling and retrograde transport. In addition, while previous studies showed that Hrs knockdown prevents EGF receptor sorting into multivesicular endosomes, we find that overexpression of SNX12 restores the sorting process in an Hrs knockdown background. Altogether, our data show that despite lower expression level, SNX12 shares redundant functions with SNX3 in the biogenesis of multivesicular endosomes

Temperature dependence of the upper critical field of an anisotropic singlet superconductivity in a square lattice tight-binding model in parallel magnetic fields

Upper critical field parallel to the conducting layer is studied in anisotropic type-II superconductors on square lattices. We assume enough separation of the adjacent layers, for which the orbital pair-breaking effect is suppressed for exactly aligned parallel magnetic field. In particular, we examine the temperature dependence of the critical field H_c(T) of the superconductivity including the Fulde-Ferrell-Larkin-Ovchinnikov (FFLO or LOFF) state, in which the Cooper pairs have non-zero center-of-mass momentum q. In the system with the cylindrically symmetric Fermi-surface, it is known that H_c(T) of the d-wave FFLO state exhibits a kink at a low temperature due to a change of the direction of q in contrast to observations in organic superconductors. It is shown that the kink disappears when the Fermi-surface is anisotropic to some extent, since the direction of q is locked in an optimum direction independent of the temperature.Comment: 5 pages, 5 figures, revtex.sty, submitted to J.Phys.Soc.Jp

Theory of de Haas-van Alphen Effect in Type-II Superconductors

Theory of quasiparticle spectra and the de Haas-van Alphen (dHvA) oscillation in type-II superconductors are developed based on the Bogoliubov-de Gennes equations for vortex-lattice states. As the pair potential grows through the superconducting transition, each degenerate Landau level in the normal state splits into quasiparticle bands in the magnetic Brillouin zone. This brings Landau-level broadening, which in turn leads to the extra dHvA oscillation damping in the vortex state. We perform extensive numerical calculations for three-dimensional systems with various gap structures. It is thereby shown that (i) this Landau-level broadening is directly connected with the average gap at H=0 along each Fermi-surface orbit perpendicular to the field H; (ii) the extra dHvA oscillation attenuation is caused by the broadening around each extremal orbit. These results imply that the dHvA experiment can be a unique probe to detect band- and/or angle-dependent gap amplitudes. We derive an analytic expression for the extra damping based on the second-order perturbation with respect to the pair potential for the Luttinger-Ward thermodynamic potential. This formula reproduces all our numerical results excellently, and is used to estimate band-specific gap amplitudes from available data on NbSe_2, Nb_3Sn, and YNi_2B_2C. The obtained value for YNi_2B_2C is fairly different from the one through a specific-heat measurement, indicating presence of gap anisotropy in this material. C programs to solve the two-dimensional Bogoliubov-de Gennes equations are available at http://phys.sci.hokudai.ac.jp/~kita/index-e.html .Comment: 16 pages, 11 figure

Magnetic-Field Variations of the Pair-Breaking Effects of Superconductivity in (TMTSF)2ClO4

We have studied the onset temperature of the superconductivity Tc_onset of the organic superconductor (TMTSF)2ClO4, by precisely controlling the direction of the magnetic field H. We compare the results of two samples with nearly the same onset temperature but with different scattering relaxation time tau. We revealed a complicated interplay of a variety of pair-breaking effects and mechanisms that overcome these pair-breaking effects. In low fields, the linear temperature dependences of the onset curves in the H-T phase diagrams are governed by the orbital pair-breaking effect. The dips in the in-plane field-angle phi dependence of Tc_onset, which were only observed in the long-tau sample, provides definitive evidence that the field-induced dimensional crossover enhances the superconductivity if the field direction is more than about 19-degrees away from the a axis. In the high-field regime for H//a, the upturn of the onset curve for the long-tau sample indicates a new superconducting state that overcomes the Pauli pair-breaking effect but is easily suppressed by impurity scatterings. The Pauli effect is also overcome for H//b' by a realization of another state for which the maximum of Tc_onset(phi) occurs in a direction different from the crystalline axes. The effect on Tc_onset of tilting the applied field out of the conductive plane suggests that the Pauli effect plays a significant role in determining Tc_onset. The most plausible explanation of these results is that (TMTSF)2ClO4 is a singlet superconductor and exhibits Fulde-Ferrell-Larkin-Ovchinnikov (FFLO) states in high fields.Comment: 12 pages, 10 figures. To be published in J. Phys. Soc. Jpn. (vol.77, 2008

The transmembrane protein p23 contributes to the organization of the Golgi apparatus

In previous studies we have shown that p23, a member of the p24-family of small transmembrane proteins, is highly abundant in membranes of the cis-Golgi network (CGN), and is involved in sorting/trafficking in the early secretory pathway, In the present study, we have further investigated the role of p23 after ectopic expression, We found that ectopically expressed p23 folded and oligomerized properly, even after overexpression, However, in contrast to endogenous p23, exogenous p23 molecules did not localize to the CGN, but induced a significant expansion of characteristic smooth ER membranes, where they accumulated in high amounts, This ER-derived, p23-rich subdomain displayed a highly regular morphology, consisting of tubules and/or cisternae of constant diameter, which were reminiscent of the CGN membranes containing p23 in control cells. The expression of exogenous p23 also led to the specific relocalization of endogenous p23, but not of other proteins, to these specialized ER-derived membranes. Relocalization of p23 modified the ultrastructure of the CGN and Golgi membranes, but did not affect anterograde and retrograde transport reactions to any significant extent. We conclude (i) that p23 has a morphogenic activity that contributes to the morphology of CGN-membranes; and (ii) that the presence of p23 in the CGN is necessary for the proper organization of the Golgi apparatus