Interactions of lipids and proteins: Some general principles

Methods to describe the binding of phospholipids to membrane proteins are described. It is shown that it is difficult to obtain estimates of the number of phospholipids bound to the surface of a membrane protein from ESR experiments in which plots of free to bound spin label (y) vs. molar ratio of lipid to protein are extrapolated toy=0. The relative advantages and disadvantages of ESR and fluorescence methods for measuring relative binding constants of phospholipids to membrane proteins are discussed. The particular problems associated with comparing binding constants of molecules of very different sizes (e.g., fatty acids and cardiolipin) are described and equations are presented to account for these problems. The possible effects of membrane viscosity and thickness on activity of membrane proteins are discussed, but it is concluded that effects of phospholipid structure on activity can only be understood in terms of a reasonably complete kinetic model for the protein.     

Some regularities of the regeneration of the musculature of internal organs]

The paper summarizes the data of the author and his co-workers on regulation of the musculature of vessels, digestive tract, uterus, ureters and urinary bladder. The smooth muscle cells have different degrees of differentiation. The muscular tissue of the vessels is most differentiated. The uterus musculature is very plastical. After injury mature myocytes are the first to undergo destruction. The intermedial substance is more stable. Myoblasts, young elements of the fibroblast row and the subendothelial layer cells are the origin of muscular regeneration. Figures of mitosis and amitosis are noted. Mature myocytes and intercellular substance are formed in the process of differentiation of the regeneration. The content of RNP in the regeneration cells is high, but in the process of differentiation of its elements it becomes lower. The DNP level has inconsiderable fluctuations. In early experiments PAS-positive substances are revealed in greater degree than in later ones. The content of acid mucopolysaccharides decreases in the process of fibrillogenesis. In all internal organs under study the muscular tissue regenerated. The degree of differentiation, severity of the lesion and functional peculiarities of the organ determine the completeness of the tissue reparation. The musculature of the intestinal tract and vessels regenerates more completely. Mighty layers of connective tissue with de novo formed blood vessels are disposed among the bundles of the repaired muscular tissue of the uterus and urine bladder wall. Simultaneously a part of regeneration cells are destroyed. These are two sides of a single process of development.     

Differential Candida albicans lipase gene expression during alimentary tract colonization and infection

The human pathogenic fungus Candida albicans, which can reside as a benign commensal of the gut, possesses a large family of lipase encoding genes whose extracellular activity may be important for colonization and subsequent infection. The expression of the C. albicans lipase gene family (LIP1-10) was investigated using a mouse model of mucosal candidiasis during alimentary tract colonization (cecum contents) and orogastric infection. LIPs4-8 were expressed in nearly every sample prepared from the cecum contents and infected mucosal tissues (stomach, hard palate, esophagus and tongue) suggesting a maintenance function for these gene products. In contrast, LIPs1, 3, and 9, which were detected consistently in infected gastric tissues, were essentially undetectable in infected oral tissues. In addition, LIP2 was expressed consistently in cecum contents but was undetectable in infected oral tissues suggesting LIP2 may be important for alimentary tract colonization, but not oral infection. The host responded to a C. albicans infection by significantly increasing expression of the chemokines MIP-2 and KC at the site of infection. Therefore, differential LIP gene expression was observed during colonization, infection and at different infected mucosal sites.     

Upregulation of telomerase function during tissue regeneration

Telomerase plays a primary role in the maintenance of telomeres in immortal, germ, and tumor cells in humans but is lacking in most somatic cells and tissues. However, many species, including fish and inbred mice, express telomerase in most cells and tissues. Little is known about the expression of telomerase in aquatic species, although the importance of telomerase for longevity has been suggested. We compared telomerase activity and telomere lengths among a broad range of tissues from aquatic species and found telomerase at significant levels in both long- and short-lived aquatic species, suggesting constitutive telomerase expression has an alternative function. Telomere lengths in these aquatic species were comparable to those observed in normal human tissues and cell strains. Given that a host of aquatic species with short life spans have telomerase and a tremendous capacity to regenerate, we tested the hypothesis that telomerase upregulation is important for tissue regeneration. During regeneration, telomerase activity was upregulated and telomere lengths are maintained with the shortest telomeres being elongated, indicating the importance for maintaining telomere length and integrity during tissue regeneration. Thus, the expression of telomerase in aquatic animals is likely not related to longevity but to their ability to regenerate injured tissue.     

The hemomicrocirculatory bed in the wall of hollow organs of the gastrointestinal tract of dogs with portal hypertension

At portal hypertension, produced by means of experimental stenosis of the portal vein in the hemomicrocirculatory bed of hollow organs of the gastrointestinal tract, congestive phenomena and edema of walls in the organs are observed. Manifested dilatation is noted in the lumen of arterioles, venules, postcapillary venules and capillaries. At early stages after the operation average diameters of these vessels in the submucosal base of the small intestine become increased 3-7 times and they do not return to the initial size even at late stages. The precapillary sphincters are in the state of spasm. Overdistention of walls in microvessels of the venular part of the functional module results in their increased permeability, that is demonstrated as diapedesic hemorrhages. During formation of intraorganic and extraorganic peripheral pathways of the circulation, the congestive phenomena in the hemomicrocirculatory bed disappear gradually.     

Alterations to the subepithelial layers of the larval alimentary tract during metamorphosis in the sea lamprey Petromyzon marinus,with emphasis on tissue interactions

Light-microscopic histochemistry and conventional electron microscopy were used to study the changes to the subepithelial layers in the larval esophagus of the sea lamprey Petromyzon marinus during metamorphosis. During early stages of metamorphosis, smooth muscle cells of the muscularis mucosae and tunica muscularis dedifferentiate into myofibroblast-like cells, which make contact with the basal lamina of the overlying mucosal epithelium. During later stages, these myofibroblast-like cells redifferentiate into smooth muscle cells, reforming the muscularis mucosae and tunica muscularis. Alterations to the extracellular matrix occur concomitantly.     

Kinins and epithelial ion transport in the alimentary tract

Kinin effects on epithelial electrogenic ion transport are reviewed, with reference to the alimentary tract. The transported ion is usually chloride, but some epithelia also transport bicarbonate. The key components of the transport system are the sodium-potassium-chloride cotransporter, Na+-K+ ATPase (both located basolaterally) and the CFTR chloride channel (located apically). Activation of K+-channels in both membranes may secondarily affect the anion transport mechanism. The types of kinin receptors that cause chloride secretion, the second messengers involved and the possible functional responsibilities of the kinin-activated secretory mechanism are discussed.     

Serotonin and neurotensin-containing cells in the mucosa of the alimentary tract

Localization of serotonin (5-OT)- and neurotensin (NT)-containing cells in the tunica mucosa of the gastric antral part and the duodenum of the rat and chick has been revealed by means of the immunofluorescent method and the method with application of the peroxidase-antiperoxidase complex. For immune-histochemical revealing of biologically active substances in endocrine cells Sovient antisera are used for the first time. Experiments are performed on injection of antiserotonin serum of various dosage into the rat blood bed. The reaction of serotonin-containing cells of the gastric antral part tunical mucosa and those of the duodenum to changes in serotonin concentration in blood serum is demonstrated.     

Composite scaffolds for the engineering of hollow organs and tissues

Several types of synthetic and naturally derived biomaterials have been used for augmenting hollow organs and tissues. However, each has desirable traits which were exclusive of the other. We fabricated a composite scaffold and tested its potential for the engineering of hollow organs in a bladder tissue model. The composite scaffolds were configured to accommodate a large number of cells on one side and were designed to serve as a barrier on the opposite side. The scaffolds were fabricated by bonding a collagen matrix to PGA polymers with threaded collagen fiber stitches. Urothelial and bladder smooth muscle cells were seeded on the composite scaffolds, and implanted in mice for up to 4 weeks and analyzed. Both cell types readily attached and proliferated on the scaffolds and formed bladder tissue-like structures in vivo. These structures consisted of a luminal urothelial layer, a collagen rich compartment and a peripheral smooth muscle layer. Biomechanical studies demonstrated that the tissues were readily elastic while maintaining their pre-configured structures. This study demonstrates that a composite scaffold can be fabricated with two completely different polymer systems for the engineering of hollow organs. The composite scaffolds are biocompatible, possess adequate physical and structural characteristics for bladder tissue engineering, and are able to form tissues in vivo. This scaffold system may be useful in patients requiring hollow organ replacement.