Skeletal muscle dispersion (400‐1000 nm) and kinetics at optical clearing

Skeletal muscle dispersion and optical clearing (OC) kinetics were studied experimentally to prove the existence of the refractive index (RI) matching mechanism of OC. Sample thickness and collimated transmittance spectra were measured during treatments with glucose (40%) and ethylene glycol (EG; 99%) solutions and used to obtain the time dependence of the RI of tissue fluids based on the proposed theoretical model. Calculated results demonstrated an increase of RI of tissue fluids and consequently proved the occurrence of the RI matching mechanism. The RI increase was observed for the wavelength range between 400 and 1000 nm and for the 2 probing molecules explored. We found that for 30 min treatment with 40% glucose and 99% EG, RI of sarcoplasm plus interstitial fluid was increased at 800 nm from 1.328 to 1.348 and from 1.328 to 1.369, respectively.      

Intracellular hypertonicity is responsible for water flux associated with Na+/glucose cotransport

Detection of a significant transmembrane water flux immediately after cotransporter stimulation is the experimental basis for the controversial hypothesis of secondary active water transport involving a proposed stoichiometry for the human Na(+)/glucose cotransporter (SGLT1) of two Na(+), one glucose, and 264 water molecules. Volumetric measurements of Xenopus laevis oocytes coexpressing human SGLT1 and aquaporin can be used to detect osmotic gradients with high sensitivity. Adding 2 mM of the substrate alpha-methyl-glucose (alphaMG) created mild extracellular hypertonicity and generated a large cotransport current with minimal cell volume changes. After 20, 40, and 60 s of cotransport, the return to sugar-free, isotonic conditions was accompanied by measurable cell swelling averaging 0.051, 0.061, and 0.077 nl/s, respectively. These water fluxes are consistent with internal hypertonicities of 1.5, 1.7, and 2.2 mOsm for these cotransport periods. In the absence of aquaporin, the measured hypertonicites were 4.6, 5.0, and 5.3 mOsm for the same cotransport periods Cotransport-dependent water fluxes, previously assumed to be water cotransport, could be largely explained by hypertonicities of such amplitudes. Using intracellular Na(+) injection and Na(+)-selective electrode, the intracellular diffusion coefficient for Na(+) was estimated at 0.29 +/- 0.03 x 10(-5) cm(2) s(-1). Using the effect of intracellular alphaMG injection on the SGLT1-mediated outward current, the intracellular diffusion coefficient of alphaMG was estimated at 0.15 +/- 0.01 x 10(-5) cm(2) s(-1). Although these intracellular diffusion coefficients are much lower than in free aqueous solution, a diffusion model for a single solute in an oocyte would require a diffusion coefficient three times lower than estimated to explain the local osmolyte accumulation that was experimentally detected. This suggests that either the diffusion coefficients were overestimated, possibly due to the presence of convection, or the diffusion in cytosol of an oocyte is more complex than depicted by a simple model.     

Simultaneous measurements of oxygen diffusion coefficients and solubilities in fermentation media with polarographic oxygen electrodes

A membrane-covered oxygen electrode was used to measure oxygen diffusion coefficients and solubilities in aqueous glucose solutions and various fermentation media following a newly developed methodology. The fermentation media studied were tryptic soy broth and those for fermentations of Penicillium chrysogenum, Saccharomyces cerevisiae, and Micrococcus glutamicus. The experimental results of oxygen diffusion coefficients and solubilities in glucose solutions were in good accord with the literature data. As for the fermentation media, both oxygen diffusion coefficients and solubilities were found to decrease with an increased fractional composition of these media, and log-additive behaviors of the oxygen diffusion coefficients and solubilities in fermentation media were observed.     

Measuring oxygen diffusion coefficients with polarographic oxygen electrodes. II. Fermentation Media

Fermentation media consist of a large number of chemicals which composition undergoes alteration during the course of fermentations. In consequence, the conventional methods and correlations for gas diffusion coefficient measurement and prediction cannot be easily applied to such systems. Oxygen diffusion coefficients have been measured in simulated chemical systems as well as in complex solutions of nutrient broth, using the polarographic technique introduced in a previous article. It is identified that sugars and salts are the major factors influencing oxygen diffusion coefficients in these aqueous fermentation media. The effect of salts on oxygen diffusion coefficients in electrolyte solutions has been found to be well correlated with the square root of total ionic strength of electrolyte solutions. The individual effect of glucose and its combined effect with salts are explored in order to reach rational correlations capable of predicting oxygen diffusion coefficients in synthetic fermentation media. For aqueous solutions of glucose plus salts, it is observed that the log-additive relationship can be used to account for the combined effect. Finally, a linear correlation has been established in measuring oxygen diffusion coefficients in aqueous solutions having different concentrations of nutrient broth.     

A robust ex vivo method to evaluate the diffusion properties of agents in biological tissues

A robust method is presented for evaluating the diffusion properties of chemicals in ex vivo biological tissues. Using this method that relies only on thickness and collimated transmittance measurements, the diffusion properties of glycerol, fructose, polypropylene glycol and water in muscle tissues were evaluated. Amongst other results, the diffusion coefficient of glycerol in colorectal muscle was estimated with a value of 3.3 × 10^?7 cm²/s. Due to the robustness and simplicity of the method, it can be used in other fields of biomedical engineering, namely in organ cryoprotection and food industry.      

Research on the Interstitial Fluid Load Support Characteristics and Start-Up Friction Mechanisms of PVA-HA-Silk Composite Hydrogel

Hydrogel has been extensively studied as an articular cartilage repair and replacement material. PVA-HA-Silk composite hydrogel was prepared by freezing-thawing method in this paper. Mechanical properties were determined by experiments and the friction coefficient of PVA-HA-Silk composite hydrogel against steel ball was verified using micro-tribometer. Finite Element Method （FEM） was used to study the lubrication mechanism of PVA-HA-Silk composite hydrogel and the relation between the interstitial fluid load support and the start-up friction resistance. The results show that the elastic modulus and the permeability are 2.07 MPa and 10^-15m^4N^-1s^-1, respectively, and the start-up friction coefficients of PVA-HA-Silk composite hydrogel are in the range of 0.154）.2 at different contact loads, contact time and sliding speeds. The start-up friction resistance of PVA-HA-Silk composite hydrogel increases with the contact load and contact time. With the increase in sliding speed, the start-up friction resistance of PVA-HA-Silk composite hydrogel decreases. There is an inverse relation between the start-up friction resistance and the interstitial fluid load support. The change of fluid flow with the increase in sliding displacement has an important effect on the interstitial fluid load support and friction resistance. The interstitial fluid load support decreases with the increase in contact load and contact time, while the interstitial fluid load support reinforces with the increase in sliding speed. Moreover, PVA-HA-Silk composite hydrogel has mechanical properties of recovery and self-lubricating.     

Relationship between Recrystallization Rate of Ice Crystals in Sugar Solutions and Water Mobility in Freeze-Concentrated Matrix

To better understand the relation between recrystallization rate and water mobility in freeze-concentrated matrix, isothermal ice recrystallization rates in several sugar aqueous solutions and self-diffusion coefficients of water component in corresponding freeze-concentrated matrix were measured. The sugars used were fructose, glucose, maltose, and sucrose. The sugar concentrations and temperature were varied so that ice contents for all samples were almost equal. Neither recrystallization rates nor diffusion coefficients depended uniformly on temperature. The recrystallization rates increased with increasing the diffusion coefficients, and a direct relationship was found between recrystallization rate and diffusion coefficient. This indicated that self-diffusion coefficient of water component in freeze-concentrated matrix is a useful parameter for predicting and controlling recrystallization rate in sugar solutions relevant to frozen desserts.     

Novel micromachined silicon sensor for continuous glucose monitoring

The construction and the application properties of a micro-machined silicon sensor for continuous glucose monitoring are presented. The sensor uses the conventional enzymatic conversion of glucose with amperometric detection of H(2)O(2). The innovation is the precise diffusion control of the analyte through a porous silicon membrane into a silicon etched cavity containing the immobilised enzyme. A variation of the number and size of the membrane pores allows to adjust the linear range of the sensor to the respective requirement. The sensor was tested in vitro as well as in clinical studies, being supplied with interstitial fluid. The cavity sensor was designed for a linear range between 0.5 and 20 mM. A signal response time of below 30 s and a signal stability exceeding 1 week is shown. By using a double cavity sensor falsification of the glucose signal by interfering substances can be compensated. In clinical trials the sensor measured continuously in interstitial fluid for up to 18 h without any signal drift and with good correlation to blood glucose reference values.     

Glucose and mannitol diffusion in human dura mater

An in vitro experimental study of the control of the human dura mater optical properties at administration of aqueous solutions of glucose and mannitol has been presented. The significant increase of the dura mater optical transmittance under action of immersion liquids has been demonstrated. Diffusion coefficients of glucose and mannitol in the human dura mater tissue at 20 degrees C have been estimated as (1.63 +/- 0.29) x 10(-6)cm(2)/s and as (1.31 +/- 0.41) x 10(-6) cm(2)/s, respectively. Experiments show that administration of immersion liquids allows for the effective control of tissue optical characteristics that make dura mater more transparent, thereby increasing the ability of light penetration through the tissue.     

Interstitial glucose concentration and glycemia: implications for continuous subcutaneous glucose monitoring

The changes in plasma glucose concentration and in interstitial glucose concentration, determined with a miniaturized subcutaneous glucose sensor, were investigated in anesthetized nondiabetic rats. Interstitial glucose was estimated through two different calibration procedures. First, after a glucose load, the magnitude of the increase in interstitial glucose, estimated through a one-point calibration procedure, was 70% of that in plasma glucose. We propose that this is due to the effect of endogenous insulin on peripheral glucose uptake. Second, during the spontaneous secondary decrease in plasma glucose after the glucose load, interstitial glucose decreased faster than plasma glucose, which may also be due to the effect of insulin on peripheral glucose uptake. Third, during insulin-induced hypoglycemia, the decrease in interstitial glucose was less marked than that of plasma glucose, suggesting that hypoglycemia suppressed transfer of glucose into the interstitial tissue; subsequently, interstitial glucose remained lower than plasma glucose during its return to basal value, suggesting that the stimulatory effect of insulin on peripheral glucose uptake was protracted. If these observations obtained in rats are relevant to human physiology, such discrepancies between plasma and interstitial glucose concentration may have major implications for the use of a subcutaneous glucose sensor in continuous blood glucose monitoring in diabetic patients.     

A theoretical study on the sucrose gap technique as applied to multicellular muscle preparations. I. Saline-sucrose interdiffusion.

Voltage-clamp analysis of membrane currents in multicellular muscle preparations by means of the sucrose gap method is complicated by diffusion of saline and sucrose in the interstitial fluid spaces. This paper is the first part of a theoretical study made to analyze electrical events related to this diffusion process. Concentration profiles of ions and sucrose (both axial and radial) were computed by solving diffusion equations with boundary conditions appropriate for the different types of preparations and experimental arrangements used. In addition to steady-state solutions, analytical expressions were derived that describe the time-course with which concentration profiles become established after a stepwise change of the solute concentration in one of the compartments of the sucrose gap apparatus. The model accounts for the presence of an endothelial surface layer, or endocardium, which acts as an external diffusion barrier and is important in determining concentration gradients of solutes within heart cell preparations. Results of numerical computations dealing with several cases of experimental interest are presented.     

Detection of glucose and related analytes by biosensors: a fractal analysis

A fractal analysis is used to model the binding and dissociation kinetics of connective tissue interstitial glucose, adipose tissue interstitial glucose, insulin, and other related analytes on biosensor surfaces. The analysis provides insights into diffusion-limited analyte-receptor reactions occurring on heterogeneous biosensor surfaces. Numerical values obtained for the binding and the dissociation rate coefficients are linked to the degree of heterogeneity or roughness (fractal dimension, Df) present on the biosensor chip surface. The binding and dissociation rate coefficients are sensitive to the degree of heterogeneity on the surface. For example, for the binding of plasma insulin, as the fractal dimension value increases by a factor of 2.47 from Df1 equal to 0.6827 to Df2 equal to 1.6852, the binding rate coefficient increases by a factor of 4.92 from k1 equal to 1.0232 to k2 equal to 5.0388. An increase in the degree of heterogeneity on the probe surface leads to an increase in the binding rate coefficient. A dual-fractal analysis is required to fit the binding kinetics in most of the cases presented. A single fractal analysis is adequate to describe the dissociation kinetics. Affinity (ratio of the binding to the dissociation rate coefficient) values are also presented. Interferents for glucose such as uric acid and ascorbic acid were also detected using glucose biosensors based on carbon nanotube (CNT) nanoelectrode ensembles (NEEs) (29) (Lin, Y.; Lu, F.; Tu, Y.; Ren, Z. Nano Lett. 2004, 4 (2), 191-195). Attempts are made to standardize biosensor properties in terms of diffusion characteristics on in vivo responsiveness.     

In vivo muscle amino acid transport involves two distinct processes

We have tested the hypothesis that transit through the interstitial fluid, rather than across cell membranes, is rate limiting for amino acid uptake from blood into muscle in human subjects. To quantify muscle transmembrane transport of naturally occurring amino acids, we developed a novel 4-pool model that distinguishes between the interstitial and intracellular fluid compartments. Transport kinetics of phenylalanine, leucine, lysine, and alanine were quantified using tracers labeled with stable isotopes. The results indicate that interstitial fluid is a functional compartment insofar as amino acid kinetics are concerned. In the case of leucine and alanine, transit between blood and interstitial fluid was potentially rate limiting for muscle amino acid uptake and release in the postabsorptive state. For example, in the case of leucine, the rate of transport between blood and interstitial fluid compared with the corresponding rate between interstitial fluid and muscle was 247 +/- 36 vs. 610 +/- 95 nmol.min(-1).100 ml leg(-1), respectively (P < 0.05). Our results are consistent with the process of diffusion governing transit from blood to interstitial fluid without selectivity, and of specific amino acid transport systems with varying degrees of efficiency governing transit from interstitial fluid to muscle. These results imply that changes in factors that affect the transit of amino acids from blood through interstitial fluid, such as muscle blood flow or edema, could play a major role in controlling the rate of muscle amino acid uptake.     

A continuous flow method for the determination of glucose uptake by excised rat diaphragm

A technique is described for the determination of the rate of uptake of glucose by the excised rat diaphragm preparation at intervals as short as 20 seconds. The rate is estimated from the fall in glucose concentration which occurs when a suitable medium flows over the muscle at constant rate. When insulin is added to the perfusion fluid, the rate of glucose uptake rises to a value about 50 per cent higher than that established before the introduction of insulin. Under the conditions described the change in uptake rate requires about 10 to 12 minutes to reach completion, being three-quarters complete in about 7 minutes. Although the major part of this delay must be ascribed to the time required for the washing out of insulin-free medium from the apparatus, and for the diffusion of insulin into the interstitial space, it is suggested that some of the delay may be due to processes occurring between the arrival of insulin at the fibre surface and the exertion of its characteristic effect on glucose uptake, although neither the time-course nor the nature of this induction phase is established.     

Brain interstitial fluid drainage and extracellular space affected by inhalational isoflurane: in comparison with intravenous sedative dexmedetomidine and pentobarbital sodium

Brain interstitial fluid drainage and extracellular space are closely related to waste clearance from the brain. Different anesthetics may cause different changes of brain interstitial fluid drainage and extracellular space but these still remain unknown. Herein,effects of the inhalational isoflurane, intravenous sedative dexmedetomidine and pentobarbital sodium on deep brain matters' interstitial fluid drainage and extracellular space and underlying mechanisms were investigated. When compared to intravenous anesthetic dexmedetomidine or pentobarbital sodium, inhalational isoflurane induced a restricted diffusion of extracellular space, a decreased extracellular space volume fraction, and an increased norepinephrine level in the caudate nucleus or thalamus with the slowdown of brain interstitial fluid drainage. A local administration of norepinephrine receptor antagonists, propranolol,atipamezole and prazosin into extracellular space increased diffusion of extracellular space and interstitial fluid drainage whilst norepinephrine decreased diffusion of extracellular space and interstitial fluid drainage. These findings suggested that restricted diffusion in brain extracellular space can cause slowdown of interstitial fluid drainage, which may contribute to the neurotoxicity following the waste accumulation in extracellular space under inhaled anesthesia per se.     

Determination of glucose diffusion coefficients in biofilms with micro-electrodes

A glucose micro-electrode was developed for direct measurements inside biofilms, and applied for the determination of effective diffusion coefficients in a model system of agar beads containing immobilized yeast cells. Two methods were used, one based on concentration gradients present at the liquid/solid interface of an active biofilm under steady-state conditions, the other based on the rate of glucose redistribution in an inactivated biofilm under transient-state conditions. Additional measurements with pH and oxygen micro-electrodes were performed and thus allowed for in-situ correction of the glucose electrode signal. From the micro-electrode measurements in the model system it was concluded that the glucose micro-sensor is a useful tool with which to obtain effective diffusion coefficients in biofilms.     

Estimation of Water Diffusion Coefficients in Freeze-Concentrated Matrices of Sugar Solutions Using Molecular Dynamics: Correlation Between Estimated Diffusion Coefficients and Measured Ice-Crystal Recrystallization Rates

The diffusion coefficient of the water component in a freeze-concentrated matrix is a useful parameter for predicting and controlling the recrystallization rate of ice crystals in sugar solutions relevant to frozen desserts. Herein, application of molecular dynamics (MD) for estimating the water diffusion coefficient in a freeze-concentrated matrix of sugar solutions is described. Diffusion coefficients evaluated using MD with the optimized potentials for liquid simulations all atom force field and water models of three types (simple point charge, simple point charge extended, and transferable intermolecular potential-4 point) show a good positive linear relation with measured values, indicating that the MD methods used in this study are useful for predicting differences in water diffusion coefficients in a sugar freeze-concentrated matrix. Furthermore, similarly to measured values, the estimated diffusion coefficients show a good positive correlation with recrystallization rates of ice crystals, which suggests that MD is useful to predict differences in recrystallization rates of ice crystals in frozen sugar solutions.     

Differences in pH between interstitial fluid and arterial blood in water-breathing and air-breathing vertebrates.

Most cells are bathed by interstitial fluid, but extracellular pH measurements are mostly for arterial plasma. Whole-body mean pH differences between the two fluids have been estimated in terms of a simple model. This relates to the diffusive exchange of carbon dioxide and oxygen and utilizes literature data, for 22 vertebrate species, on arterial and mixed-venous tensions of both gases. Uncertainties arise because the carbon dioxide reaction in blood may sometimes be in disequilibrium and because carbon dioxide diffusion is facilitated to unknown degrees in the presence of buffers. Nevertheless, the model suggests that the pH difference should tend to vary inversely with arterial carbon dioxide tension. In some species, this may aid interstitial pH homeostasis, but a clearer implication is that the difference should be generally greater in water breathers than in air breathers. It has previously been found that arterial pH in water-breathing teleosts also tends to be higher than in air-breathing tetrapods (when allowance is made for temperature and plasma sodium concentration) and to a comparable extent. Thus, mean interstitial pH may be more nearly similar in the two groups than is arterial pH. Direct measurements of interstitial pH do not yet suffice to test the model.     

An approximation method for the determination of diffusion and permeability coefficients in nerve trunks

A recently introduced approximation method is applied in order to obtain an expression for the amount of a substance remaining within a nerve at any time, the nerve having been soaked for a long time in a solution containing the substance until the time zero when it is bathed in the same solution but without the substance. The case of a uniform nerve without a sheath leads to substantially the same results as previously obtained by A. V. Hill (1928) for this case. A solution is given for the case of a nerve without sheath but having fibers which are permeable. In this case it is shown how an effective diffusion coefficient for the interstitial fluid can be obtained, as well as the effective inward and outward fiber permeabilities. A solution is given for the case of a nerve with a sheath in which the substance considered does not penetrate the fibers, and it is shown how the effective diffusion coefficients of the sheath and the interstitial fluid can be obtained.     

Distribution of saccharides and salts on amphoteric ion-exchange resin

An amphoteric ion-exchange resin hardly shrank in 550 and 300 g/L glucose and sodium chloride solutions, respectively; however, the bed packed with a cation-exchange resin shrank considerably. From the distribution coefficients of some saccharides, the swelling pressure of the amphoteric ion-exchange resin was estimated to be 2.0 MPa at 25 °C. The distribution coefficients of glucose, galactose, fructose, and mannose were independent of their concentration and were about 0.621. On the other hand, the apparent distribution coefficients of NaF, NaCl, NaBr, NaI, LiCl, KCl, and CsCl largely depended on concentration. A model for the distribution of salts on the amphoteric resin was proposed, assuming an interaction between the anion of the salt and the positively charged fixed ions with binding constant B. The B values of the chloride salts were nearly the same (1.69–2.94 L/mol), while the values of the sodium salts were largely different depending on the anion.