An endosomal beta COP is involved in the pH-dependent formation of transport vesicles destined for late endosomes

In this paper, we show that beta COP is present on endosomes and is required for the formation of vesicles which mediate transport from early to late endosomes. Both the association of beta COP to endosomal membranes as well as transport vesicle formation depend on the lumenal pH. We find that epsilon COP, but not gamma COP, is also associated to endosomes, and that this association is also lumenal pH dependent. Our data, thus, indicate that a subset of COPs is part of the mechanism regulating endosomal membrane transport, and that membrane association of these COPs is controlled by the acidic properties of early endosomes, presumably via a trans-membrane pH sensor

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

Vertebral Artery Injury: Case Report and Review of Operative Approaches

Traumatic injury to the upper third of the neck, cephalad to the angle of the mandible, distorts the complexity of normal anatomy and underscores the importance of preoperative arteriography when possible. This report describes the case of a young man who was brought to the Emergency Room of our hospital suffering from a severe gunshot wound to the cervical spine, vertebral artery, and maxillofacial skeleton. His injury was successfully managed by combining a standard arterolateral incision with a procedure that has been described for exposing retro maxillary tumors. This operative technique provided the surgeon with direct access to the injury, controlled the loss of blood, and permitted repair of accompanying pharyngeal and facial injuries. Our report also reviews the technical considerations and pertinent surgical anatomy of this rare combination of Injuries

The trans-membrane protein p25 forms highly specialized domains that regulate membrane composition and dynamics

Trans-membrane proteins of the p24 family are abundant, oligomeric proteins predominantly found in cis-Golgi membranes. They are not easily studied in vivo and their functions are controversial. We found that p25 can be targeted to the plasma membrane after inactivation of its canonical KKXX motif (KK to SS, p25SS), and that p25SS causes the co-transport of other p24 proteins beyond the Golgi complex, indicating that wild-type p25 plays a crucial role in retaining p24 proteins in cis-Golgi membranes. We then made use of these observations to study the intrinsic properties of these proteins, when present in a different membrane context. At the cell surface, the p25SS mutant segregates away from both the transferrin receptor and markers of lipid rafts, which are enriched in cholesterol and glycosphingolipids. This suggests that p25SS localizes to, or contributes to form, specialized membrane domains, presumably corresponding to oligomers of p25SS and other p24 proteins. Once at the cell surface, p25SS is endocytosed, together with other p24 proteins, and eventually accumulates in late endosomes, where it remains confined to well-defined membrane regions visible by electron microscopy. We find that this p25SS accumulation causes a concomitant accumulation of cholesterol in late endosomes, and an inhibition of their motility - two processes that are functionally linked. Yet, the p25SS-rich regions themselves seem to-exclude not only Lamp1 but also accumulated cholesterol. One may envision that p25SS accumulation, by excluding cholesterol from oligomers, eventually overloads neighboring late endosomal membranes with cholesterol beyond their capacity (see Discussion). In any case, our data show that p25 and presumably other p24 proteins are endowed with the intrinsic capacity to form highly specialized domains that control membrane composition and dynamics. We propose that p25 and other p24 proteins control the fidelity of membrane transport by maintaining cholesterol-poor membranes in the Golgi complex

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.

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

Superconducting fluctuations at low temperature

The effect of fluctuations on the transport and thermodynamic properties of two-dimensional superconductors in a magnetic field is studied at low temperature. The fluctuation conductivity is calculated in the framework of the perturbation theory with the help of usual diagram technique. It is shown that in the dirty case the Aslamazov-Larkin, Maki-Thomson and Density of States contributions are of the same order. At extremely low temperature, the total fluctuation correction to the normal conductivity is negative in the dirty limit and depends on the external magnetic field logarithmically. In the non-local clean limit, the Aslamazov-Larkin contribution to conductivity is evaluated with the aid of the Helfand-Werthamer theory. The longitudinal and Hall conductivities are found. The fluctuating magnetization is calculated in the one-loop and two-loop approximations.Comment: 12 pages, 4 figures, submitted to Phys. Rev.

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