Effect of membrane stress on surface roughness changes in sheet forming

Friction plays an important role in sheet metal forming (SMF) and the roughness of the surface of the sheet is a major factor that influences friction. In finite element method (FEM) models of metal forming, the roughness has usually been assumed to be constant; even though it is commonly observed that sheet drawn under tension over a tool radius results in the surface becoming shiny, indicating a major change in surface morphology. An elastic–plastic FEM model for micro-contact between a flat surface and a single roughness peak has been developed. The model was used to investigate the effect of the membrane stress in the sheet on the deformation of an artificial roughness peak. From the simulation results, the change in asperity, or deformation of the local peak, for a given nominal tool contact stress is significantly influenced by the local substrate stress. The height of the asperity decreases with increasing substrate stress and the local pressure is much higher than the nominal pressure. In addition, the local contact stress decreases with an increase in the substrate stress levels.

A novel polymer membrane for extraction applications

In this study, a new type of Aliquat 336/PVC membrane has been made for extraction experiments. This new membrane is capable of holding more Aliquat 336 than previously developed extraction membranes, hence overcoming a major problem that has confronted many researchers for a long time. The new membrane has been used to investigate the rate of extraction for the Cd(II) ion in 2.0 M HCl solution and the effect of membrane thickness on the rate of extraction. The experimental results have shown this new membrane has a promising future in relevant industrial applications. A new method is also used in this study to qualitatively identify the oily substance on the surface of membrane after the extraction experiment was completed. This oily substance has been found to be Aliquat 336.

Identification of an integral plasma membrane pool of protein kinase C in mammalian tissues and cells

Protein kinase C (PKC) is a family of serine/threonine protein kinases that are pivotal in cellular regulation. Since its discovery in 1977, PKCs have been known as cytosolic and peripheral membrane proteins. However, there are reports that PKC can insert into phospholipids vesicles in vitro. Given the intimate relationship between the plasma membrane and the activation of PKC, it is important to determine whether such “membrane-inserted” form of PKC exists in mammalian cells or tissues. Here, we report the identification of an integral plasma membrane pool for all the 10 PKC isozymes in vivo by their ability to partition into the detergent-rich phase in Triton X-114 phase partitioning, and by their resistance to extractions with 0.2 M sodium carbonate (pH 11.5), 2 M urea and 2 M sodium chloride. The endogenous integral membrane pool of PKC in mouse fibroblasts is found to be acutely regulated by phorbol ester or diacylglycerol, suggesting that this pool of PKC may participate in cellular processes known to be regulated by PKC. At least for PKCα, the C2–V3 region at the regulatory domain of the kinase is responsible for membrane integration. Further exploration of the function of this novel integral plasma membrane pool of PKC will not only shed new light on molecular mechanisms underlying its cellular functions but also provide new strategies for pharmaceutical modulation of this important group of kinases.

Pituitary adenylate cyclase-activating polypeptide induces translocation of its G-protein-coupled receptor into caveolin-enriched membrane microdomains, leading to enhanced cyclic AMP generation and neurite outgrowth in PC12 cells

Pituitary adenylate cyclase-activating polypeptide (PACAP), a member of the secretin/glucagon/vasoactive intestinal peptide family expressed throughout the nervous system, binds to the PACAP-specific G-protein-coupled receptor family members to promote both neuronal differentiation and survival. Although the PACAP receptor is known to activate its effector protein, adenylate cyclase (AC), and thus enhance cAMP generation, the molecular mechanism utilized by the receptor to activate AC is lacking. Here, we show that PACAP induces neurite outgrowth in PC12 cells by induction of translocation of the PACAP type 1 receptor (PAC1R) into caveolin-enriched Triton X-100-insoluble microdomains, leading to stronger PAC1R-AC interaction and elevated cAMP production. Moreover, we demonstrate that translocation of PAC1R is blocked by various treatments that selectively disrupt caveolae. As a result, intracellular cAMP level is decreased and consequently the PACAP-induced neurite outgrowth retarded. In contrast, addition of exogenous ganglioside GM1 to the cells shows the opposite effects. These results therefore identify the PACAP-induced translocation of its G-protein-coupled receptor into caveolae, where both AC and the regulating G-proteins reside, as the key molecular event in activating AC and inducing cAMP-mediated differentiation of PC12 cells.

Purification and membrane reconstitution of catalytically active Menkes copper-transporting P-type ATPase (MNK; ATP7A)

The MNK (Menkes disease protein; ATP7A) is a major copper- transporting P-type ATPase involved in the delivery of copper to cuproenzymes in the secretory pathway and the efflux of excess copper from extrahepatic tissues. Mutations in the MNK (ATP7A) gene result in Menkes disease, a fatal neurodegenerative copper deficiency disorder. Currently, detailed biochemical and biophysical analyses of MNK to better understand its mechanisms of copper transport are not possible due to the lack of purified MNK in an active form. To address this issue, we expressed human MNK with an N-terminal Glu-Glu tag in Sf9 [Spodoptera frugiperda (fall armyworm) 9] insect cells and purified it by antibody affinity chromatography followed by size-exclusion chromatography in the presence of the non-ionic detergent DDM (n-dodecyl b-D-maltopyranoside). Formation of the classical vanadate-sensitive phosphoenzyme by purified MNK was activated by Cu(I) [EC50=0.7 µM; h (Hill coefficient) was 4.6]. Furthermore, we report the first measurement of Cu(I)-dependent ATPase activity of MNK (K0.5=0.6 µM; h=5.0). The purified MNK demonstrated active ATP-dependent vectorial 64Cu transport when reconstituted into soya-bean asolectin liposomes. Together, these data demonstrated that Cu(I) interacts with MNK in a co-operative manner and with high affinity in the sub-micromolar range. The present study provides the first biochemical characterization of a purified full-length mammalian copper-transporting P-type ATPase associated with a human disease.

The Omp85 family of proteins is essential for outer membrane biogenesis in mitochondria and bacteria

Integral proteins in the outer membrane of mitochondria control all aspects of organelle biogenesis, being required for protein import, mitochondrial fission, and, in metazoans, mitochondrial aspects of programmed cell death. How these integral proteins are assembled in the outer membrane had been unclear. In bacteria, Omp85 is an essential component of the protein insertion machinery, and we show that members of the Omp85 protein family are also found in eukaryotes ranging from plants to humans. In eukaryotes, Omp85 is present in the mitochondrial outer membrane. The gene encoding Omp85 is essential for cell viability in yeast, and conditional omp85 mutants have defects that arise from compromised insertion of integral proteins like voltage-dependent anion channel (VDAC) and components of the translocase in the outer membrane of mitochondria (TOM) complex into the mitochondrial outer membrane.

Reverse osmosis membrane fouling and its control

Reverse osmosis (RO) membranes can be used to treat ground water, surface water and wastewater, but a major problem in RO membrane applications is fouling. Many studies have investigated the mechanisms of fouling in RO membranes and suggested various methods to control the membrane fouling. This paper reviews several aspects of RO membranes, such as the types of membrane fouling, their effects on RO membrane processes and fouling control technologies, focussed on the appropriate feed pretreatment technologies and cleaning methods, especially on the membrane filtration pretreatment processes, including microfiltration (MF), ultrafiltration (UF) and nanofiltration (NF), which are the new trends in designing a pretreatment system in the application of RO.

Characterization of a distinct plasma membrane macrodomain in differentiated adipocytes.

Caveolae are small invaginations of the cell surface that are abundant in mature adipocytes. A recent study (Kanzaki, M., and Pessin, J. E. (2002) J. Biol. Chem. 277, 25867-25869) described novel caveolin- and actin-containing structures associated with the adipocyte cell surface that contain specific signaling proteins. We have characterized these structures, here termed "caves," using light and electron microscopy and observe that they represent surface-connected wide invaginations of the basal plasma membrane that are sometimes many micrometers in diameter. Rather than simply a caveolar domain, these structures contain all elements of the plasma membrane including clathrin-coated pits, lipid raft markers, and non-raft markers. GLUT4 is recruited to caves in response to insulin stimulation. Caves can occupy a significant proportion of the plasma membrane area and are surrounded by cortical actin. Caveolae density in caves is similar to that on the bulk plasma membrane, but because these structures protrude much deeper into the plane of focus of the light microscope molecules such as caveolin and other plasma membrane proteins appear more concentrated in caves. We conclude that the adipocyte surface membrane contains numerous wide invaginations that do not represent novel caveolar structures but rather large surface caves.

Micro- and nano-characterization of membrane materials

The global warming has led to enormous challenges worldwide for a large variety of communities, particularly these associated to water and water treatment industry. Due to increasing scarcity of water resources, the development of new membrane materials and water treatment processes will play an important role in tackling this emerging problem. In this paper, the recent development in characterization of in particular, geometrical parameters of micro- and nano-membrane materials will be reviewed. Membranes with micro- and nano-pores have widely been used for ultrafiltration and nanofiltration. The structure of the pores and the surface of the pores/membranes may be optimized to achieve much improved flow rate in these micro-/nano-channels. Therefore, accurate characterization of porous structures will contribute significantly to the prediction of membrane performance. It will not only provide an insight into the new characterization methods but also the development of novel materials.

Examination of the role of intestinal Fatty acid-binding protein in drug absorption using a parallel artificial membrane permeability assay

Transcellular diffusion across the absorptive epithelial cells (enterocytes) of the small intestine is the main route of absorption for most orally administered drugs. The process by which lipophilic compounds transverse the aqueous environment of the cytoplasm, however, remains poorly defined. In the present study, we have identified a structurally diverse group of lipophilic drugs that display low micromolar binding affinities for a cytosolic lipid-binding protein—intestinal fatty acid-binding protein (I-FABP). Binding to I-FABP significantly enhanced the transport of lipophilic drug molecules across a model membrane, and the degree of transport enhancement was related to both drug lipophilicity and I-FABP binding affinity. These data suggest that intracellular lipid-binding proteins such as I-FABP may enhance the membrane transport of lipophilic xenobiotics and facilitate drug access to the enterocyte cytoplasm and cytoplasmic organelles.

3-dimensional characterization of membrane with nanoporous structure using TEM tomography and image analysis

The nanoporous structure of membrane varies in 3-dimensional (3-D) space and has remarkable influences on the filtration or desalination achieved, fouling potentials and therefore, the quality of yielded water. Knowledge of the 3-D nanoporous structure is thus vital to understanding and predicting its performance. A novel method by incorporating transmission electronic microtomography, image processing and 3-D reconstruction is introduced to characterize membranes with nano structures. The reconstruction algorithm allows for the visualization of 3-D nanoporous structure in a non-destructive way from any directions. This novel technique Ieads to in-depth understanding and accurate prediction of filtration performance.