Dependence of the apparent thickness of the unstirred layer at the intestinal mucosa on nutrient concentration

The thickness of the intestinal unstirred layer determined with glycine is shown to increase from 247 ± 12 to 319 ± 14 μm (p < 0.001) as its concentration is raised from 5 to 40 mM; with glucose raised from 10 to 40 mM, the apparent value increases from 316 ± 13 to 380 ± 26 μm, because at higher concentrations the nutrients penetrate deeper to the bases of intestinal villi. The weak positive correlation between the apparent layer thickness (assayed with glucose, maltose, sucrose, alanine, leucine, and Gly-Ala in rats) and the parameter characterizing the rate of nutrient uptake (maximal short-circuit current response to nutrient) contradicts the assertion that the transporters are distributed uniformly along the villus height but because of high absorption in the upper part the nutrients do not reach the deeper transporters. It appears that the transporter distribution along the villi is nonuniform. The described method may serve to study the topography of nutrient transporters.     

Measurement of sodium ion concentration in the unstirred layer of rat small intestine by polymer Na+-sensitive electrodes

[14C]Formate is incorporated into the C-2 of the pyrimidine moiety of thiamin by Escherichia coli and Salmonella typhimurium. In Saccharomyces cerevisiae, it is incorporated into C-4. Radioactive carbons of [1-14C]glycine and [2-14C]glycine are incorporated by S. typhimurium into the C-4 and C-6 of the pyrimidine, respectively, but not by S. cerevisiae. These facts suggest that procaryotes and eucaryotes have different biosynthetic pathways for pyrimidine. In this study, the procaryotes tested incorporated [14C]formate into the C-2 and the eucaryotes incorporated it into the C-4 of the pyrimidine.     

Effects of azide on gastric mucose

Sodium azide, a classical inhibitor of cytochrome oxidase, is an effective inhibitor of gastric acid secretion in bullfrog and skate gastric mucosae at low concentrations. While a portion of the oxygen uptake in these tissues is sensitive to azide (KI less than 2 mM), there remains a large fraction (25-60%) with a KI more than 10 times this value, suggesting the presence of a second oxidase. The spectra of cytochromes c and b change with oxygen-nitrogen alternation in the presence of high azide concentrations which essentially eliminate the reactivity of cytochrome oxidase. In both species two additional components are observed in the spectra. The first has a peak at 590 nm, is not the cytochrome oxidase-CO complex, is fully reactive in the presence of azide and accounts for the asymmetry of the oxidase peak. The second is a component at 557 nm which can only be separated from cytochromes c and b by spectral deconvolution, and seems to react in a manner similar to cytochrome c. It is suggested that the 590 compound may be the alternate cytochrome oxidase.     

Derivation of the equations that describe the effects of unstirred water layers on the kinetic parameters of active transport processes in the intestine

Unidirectional flux of solutes into the intestinal mucosal cells is determined by the rate of movement of these molecules across both an unstirred water layer and the microvillus membrane of the epithelial cell. Therefore, an equation is derived in this paper that describes the velocity of active transport as a function of the characteristics of both the transport carrier in the membrane and the resistance of the overlying unstirred water layer. Using this equation a series of curves are presented that depict the effect on the kinetics of active transport of varying the thickness (d) or surface area (S_w) of the unstirred water layer, the free diffusion coefficient (D) of the solute, the distribution of active transport sites along the villus ($\text{?}{}_{\mathrm{<mtext>n</mtext>}}$), the maximal transport velocity (J^m_d) and the true Michaelis constant (K_m). These theoretical curves illustrate the serious limitations inherent in interpretation of previously published data dealing with active transport processes in the intestine.     

Effects of unstirred layers or transport number discontinuities on the transient and steady-state current-voltage relationships of membranes

The effects of current-induced electrolyte accumulation and depletion on the electrical properties of a two-layered membrane system have been examined. The membrane consisted of a charged, ion permselective layer and an uncharged, non-selective layer. The model was designed to reveal the properties of membranes possessing long pores with ionic charges at one end or of ion-selective membranes bounded by highly unstirred aqueous layers. Electrolyte concentration profiles in the inert layer and their time-dependent changes were obtained from solutions of the diffusion equation under the condition of constant current. The profiles were then used to calculate the voltage developed across the membrane at various times after the current is switched on. The theoretical results are presented in the form of $\text{i-V}$ curves with reduced coordinates that can be used to obtain time-current-voltage relationships for membranes of the type considered having any thickness of the non-selective layer and bathed in any concentration of any 1:1 electrolyte. Experimental results on a model composite membrane were in good agreement with calculations that assume that ion transport occurs only under the influence of electrical potential and concentration gradients, suggesting that in such systems, the combined effects of convection, osmosis, electro-osmosis, and concentration-dependence of diffusion coefficients, activity coefficients, and transference numbers are small. Voltage fluctuations in the form of periodic spikes were observed experimentally at the limiting current density (the current density at which the electrolyte concentration at one surface of the selective layer goes to 0). These phenomena were not seen when the current was in the direction leading to accumulation of electrolyte in the non-selective (unstirred) layer. Such composite membranes can exhibit S-shaped and N-shaped $\text{i-V}$ curves under ramp-voltage and ramp-current clamps, respectively.     

Micromolar concentration of kynurenic acid in rat small intestine

Kynurenic acid is an antagonist of glutamate and alpha-7 nicotinic acetylcholine receptors and an agonist of the G: -protein-coupled receptor GPR35, which is predominantly expressed in immune and gastrointestinal tissues. In this study, we report that kynurenic acid is present in the lumen of rat small intestine in micromolar concentration sufficient to affect the GPR35 receptor. Moreover, we show that kynurenic acid can be produced by Escherichia coli. We suggest that kynurenic acid may modulate gastrointestinal function and integrity.     

Characterization of macrophage-like cells in the external layers of human small and large intestine

Summary In the external layers of human small and large intestine macrophage-like cells were characterized by immunohistochemical, histochemical and electronmicroscopical methods. Using immunohistochemistry and a number of monoclonal antibodies, the presence and distribution of phenotypic subpopulations of macrophages were evaluated. In all locations macrophage-like cells were identified with antibody EBM11, which recognizes CD68 antigen, C3bi which recognizes CD11b, and partly with an antibody which recognizes protein 150,95 (CD11c). Macrophage-like cells in the external muscle layer were HLA-DR-positive (expressing the MHC class-II antigen), in contrast to macrophage-like cells in the subserosa and submucosa. Macrophage-like cells in the external muscle layer were mostly acid phosphatase-negative, and at the electron-microscopic level they were found to have features of macrophages: primary lysosomes, coated vesicles and pits. However, very few secondary lysosomes were present. Birbeck granules were not observed. It is concluded that in the external muscle layer of human small and large intestine numerous macrophages of a special type are present. It is discussed whether this cell type plays a role in gastrointestinal motility and/or has an immunological function.     

Effects of serotonin on the physiology of the rabbit small intestine

Serotonin has been shown to alter the intestinal transport of ions and intestinal motility. These effects may interfere with each other, modulating the whole physiology of the intestine. We have previously shown that serotonin also alters the transport of nutrients. Thus, the aims of the present work were to determine the possible interference between the secretagogue effect of serotonin and the mechanism by which serotonin inhibits the absorption of nutrients, and to study the effect of serotonin on the digestive activity of nutrients of the brush border membrane jejunum enterocyte in the rabbit. The results show that the secretagogue effect of serotonin neither affects the inhibitory effect of serotonin on the intestinal absorption of the nutrients, nor affects the activity of Na+/K+-ATPase. The activity of sucrase and aminopeptidase N was also not affected by serotonin in the rabbit jejunum. Finally, we also studied different parameters of the motility in the rabbit small intestine. Serotonin seemed to stimulate the motility of the rabbit small intestine by increasing integrated mechanical activity and tone of muscle fibers in duodenum, jejunum, and ileum. In conclusion, serotonin might alter or modulate the whole intestinal physiology.     

Alterations in electrophysiology of isolated amphibian small intestine produced by removing the muscle layers

Isolated segments of Amphiuma small intestine bathed in chloride or sulfate buffer generate a greater short-circuit current and a larger change in current in response to galactose when the serosal muscle layers are stripped from the mucosa. Intact (unstripped) segments are not apparently anoxic since stripped segments exposed to serosal N₂ for 3 h display normal short-circuit currents but a reduced potential response to galactose, while the presence of muscle layers tends to reduce the short-circuit current but does not alter the potential response to galactose. Bullfrog small intestine also generates greater short-circuit current following removal of the muscle layers. The enhancing effect of stripping appears to be related to removal of a resistance to ion flow across the tissue.     

Estimation of thickness of concentration boundary layers by osmotic volume flux determination

The estimation method of the concentration boundary layers thicknesses (δ) in a single-membrane system containing non-electrolytic binary or ternary solutions was devised using the Kedem-Katchalsky formalism. A square equation used in this method contains membrane transport (L(p), σ, ω) and solution (D, C) parameters as well as a volume osmotic flux (J(v)). These values can be determined in a series of independent experiments. Calculated values δ are nonlinearly dependent on the concentrations of investigated solutions and the membrane system configuration. These nonlinearities are the effect of a competition between spontaneously occurring diffusion and natural convection. The mathematical model based on Kedem-Katchalsky equations and a concentration Rayleigh number (R(C)) was presented. On the basis of this model we introduce the dimensionless parameter, called by us a Katchalsky number (Ka), modifies R(C) of membrane transport. The critical value of this number well describes a moment of transition from the state of diffusion into convective diffusion membrane transport.     

Effects of ketohexosemia on the ketohexose transport in the small intestine of rats

It was observed previously (Csáky, T.Z. and Fischer, E. (1981) Diabetes 30, 568–574), that sustained hyperglycemia enhances the intestinal transport of aldohexoses; on the other hand, hyperfructosemia affects primarily the transport of fructose. The present study examines in detail the hyperketosemia-induced intestinal ketose transport. Intravenously infused $\text{3-O-}\text{methylfructose}$ produces marked $\text{3-O-}\text{methylfructosemia}$ without concomitant hyperglycemia; in such animals the intestinal transport of both fructose and $\text{3-O-}\text{methylfructose}$ increased. The hyperketosemia-induced increased ketose transport was inhibited by phloretin but only if placed on the serosal compartment. Phlorizin affects neither the basal nor the induced intestinal ketohexose transport. The enhancement of the intestinal ketohexose transport is not sodium-dependent and is not inhibited by ouabain.     

Direct measurement of the thickness of the unstirred diffusion layer outside immobilized biocatalysts

The concentration gradient of O₂ outside cells and oxygen-consuming particles and fibres with immobilized glucose oxidase (EC 1.1.3.4) has been directly measured with oxygen microelectrodes. Measurements have been performed in systems with different relative velocities between particles and solvent. In systems with a relative velocity equal to zero (unstirred systems), the thickness of the diffusion layer was found to be approximately the radius of the particles, and much greater than the radius of the fibres. This is in agreement with the results predicted by theoretical analysis. The thickness of the diffusion layer was not negligible, even at the highest relative velocity (1.5 x 10⁻⁴ ms⁻¹ used in this study.     

Effect of unstirred layers on binding and reaction kinetics at a membrane surface

The effects of solution unstirred layers on the time course of chemical reactions and transport processes at a membrane surface are determined. A set of equations which describes non-steady-state diffusion through an unstirred layer coupled with chemical reaction at a membrane surface or transport through a membrane is developed. A numerical solution to the equations is obtained by uncoupling diffusive and chemical processes in an iterative manner. The diffusive process is solved by the Crank-Nicolson method; the chemical process is solved by integrating the differential equations describing the kinetics. Diffusive processes in one dimension, in three dimensions, and in the presence of an arbitrary potential near the membrane surface are solved. General characteristics of the calculated reaction time course are discussed using surface binding and membrane transport examples. Small, neglected, unstirred layers are shown to sometimes yield erroneous values of rate parameters for a surface reaction and to simulate competitive reaction kinetics. Experimental approaches for measuring unstirred layer thickness are reviewed.     

Effects of whole-stream nutrient enrichment on the concentration and abundance of aquatic hyphomycete conidia in transport

The concentrations and relative abundances of aquatic hyphomycete conidia in water were followed during a three-year study in two headwater streams at Coweeta Hydrologic Laboratory, North Carolina, using the membrane-filtration technique. After a one-year pretreatment period, one of the streams was enriched continuously with inorganic nutrients (N+P) for two years while the other stream served as the reference. This ecosystem-level nutrient manipulation resulted in concentrations of aquatic hyphomycete conidia in the water of the treated stream that were 4.5-6.9 times higher than the concentrations observed during the pretreatment period and in the reference stream. Nutrient enrichment led to an increase in the number of fungal species detected on each sampling date. Changes in dominance patterns and relative abundances of individual species also were detected after treatment. Nutrient addition stimulates the reproductive activity of aquatic hyphomycetes, their colonization success and fungal-mediated leaf-litter decomposition. Such changes in the activity of the fungal community might affect higher trophic levels in lotic ecosystems.