Cellular polarization

Electron microscopic evaluation of infected Caco-2 monolayers grown on membranes, under conditions that induced cellular polarization, prompted the conclusion that the larvae occupy the cytoplasm of cells they invade (ManWarren et al., 1997). Apical and basal plasma membranes appeared to be preserved in infected cells (Fig. 6.3). These findings reproduced the observations of Wright (1979) in his examination of intestinal tissues from infected mice. In contrast, when Li et al. (1998) performed similar experiments in HT29 monolayers grown on plastic, they concluded... 

Ohno, S. 2001. Intercellular junctions and cellular polarity the PAR-aPKC complex, a conserved core cassette playing fundamental roles in cell polarity. Curr. Opin. Cell Biol. 13 641-648. 

When Na influx is blocked by TTX, activation of the AHP is preceded by a regenerative Ca potential and thus the sensitivity or threshold of the AHP to Ca determines the extent to which Ca " contributes to cellular polarization. If for instance, the Ca " threshold is very low, Ca mediated depolarizations would be followed immediately by rapid repolarization. 

Another approach to the study of membrane asymmetry has been based on the morphological and functional asymmetry of cells from organized tissues. Cellular polarity is expressed in the diverse biochemistry of the various membrane portions and refinements in plasma membrane isolation procedures have met with considerable success in ascribing membrane elements to topographical aspects of the cell. For instance, hepatocyte bile front has been separated from the sinus front and the contiguous membrane (EVANS 6e GURD,... 

Another feature of tumor cells, particularly those with high malignancy, is the loss of cellular polarity. Normal, non-transformed cells, with the exception of blood cells, have a clearly polarized structure i) surface membrane structure involved in cell-to-cell or cell-to-extracellular matrix adhesion ii) surface structure open to the external environment and involved in endocytosis or exocytosis. Structure (i) in epithelial cells, termed basolateral surface, is rich in adhesion receptors including integrins (ii), termed apical surface, has much higher level of sphingolipids,... 

A process in which a substance gains entry into a cell. Endocytic mechanisms are crucial for a variety of cellular functions such as the uptake of nutrients, regulation of cell surface expression of receptors, maintenance of cell polarity, and more. Receptor-mediated endocytosis via clathrin-coated pits is the most studied endocytic process, which is important for regulation of the time and magnitude of signals generated by a variety of cell-surface receptors. 

Microtubules are universally present in eukaryotes from protozoa to the cells of higher animals and plants (Porter, 1966 Hardham and Gunning, 1978 Lloyd, 1987), but they are absent in mammalian erythrocytes and in prokaryotes. Microtubules participate in a number of cellular functions including the maintenance of cell shape and polarity, mitosis, cytokinesis, the positioning of organelles, intracellular transport to specific domains, axoplasmic transport, and cell locomotion. The diversity of microtubule fimctions suggests that not all microtubules are identical and that different classes of microtubules are present in different cell types or are localized in distinct domains in the same cell type (Ginzburg et al., 1989). 

In this section we describe a cellular automata model of a semipermeable membrane separating two compartments [5]. A solute in one compartment has varied parameters to reflect its relative polarity or lipophilicity. The passage of this solute into and through the membrane is observed, as this property is varied. 

As with urine, saliva (spumm) is easy to collect. The levels of protein and lipids in saliva or spumm are low (compared to blood samples). These matrices are viscous, which is why extraction efficiency of xenobioties amoimts to only 5 to 9%. By acidifying the samples, extraction efficiencies are improved as the samples are clarified, and proteinaceous material and cellular debris are precipitated and removed. Some xenobioties and their metabohtes are expressed in hair. Hair is an ideal matrix for extraction of analytes to nonpolar phases, especially when the parent xenobioties are extensively metabolized and often nondetectable in other tissues (parent molecules of xenobioties are usually less polar than metabolites). Hair is a popular target for forensic purposes and to monitor drug compliance and abuse. Human milk may be an indicator of exposure of a newborn to compounds to which the mother has been previously exposed. The main components of human milk are water (88%), proteins (3%), lipids (3%), and carbohydrates in the form of lactose (6%). At present, increasing attention is devoted to the determination of xenobioties in breath. This matrix, however, contains only volatile substances, whose analysis is not related to PLC applications. 

While many biological molecules may be targets for oxidant stress and free radicals, it is clear that the cell membrane and its associated proteins may be particularly vulnerable. The ability of the cell to control its intracellular ionic environment as well as its ability to maintain a polarized membrane potential and electrical excitability depends on the activity of ion-translocating proteins such as channels, pumps and exchangers. Either direct or indirect disturbances of the activity of these ion translocators must ultimately underlie reperfiision and oxidant stress-induced arrhythmias in the heart. A number of studies have therefore investigated the effects of free radicals and oxidant stress on cellular electrophysiology and the activity of key membrane-bound ion translocating proteins. 

Several extraction techniques have also been described that use enzymatic or chemical reactions to improve extraction efficiency. A technique that has been used to increase the overall recovery of the marker residue is enzymatic hydrolysis to convert specific phase II metabolites (glucuronides or sulfates) back into the parent residue. Cooper etal used a glucuronidase to increase 10-fold the concentration of chloramphenicol residues in incurred tissue. As an example of a chemical reaction, Moghaddam et al. used Raney nickel to reduce thioether bonds between benomyl and polar cellular components, and as a result achieved a substantially improved recovery over conventional solvent extraction. In choosing to use either of these approaches, thorough characterization of the metabolism in the tissue sample must be available. 

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