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Polypropylene hollow fiber for cells isolation: methods for evaluation of diffusive transport and quality of cells encapsulation.

Formulation of membrane properties is important prior the successful implantation of encapsulated cells producing therapeutically relevant compounds. The purpose of our study was to specify the methods allowing preliminary evaluation of hollow fibers (HF) chosen for immunoisolation. We have selected as estimates (1) diffusive permeability for small and large solutes, and HF cut off (in vitro), (2) histological evaluation of tissue overgrowth after sc. implantation into mice. It was found that diffusive coefficients were linearly dependent on the particle diameter except that of albumin (2-3 times higher than theoretically estimated). This discrepancy imply that for certain particles the interaction with membrane material may be significant. The histological evaluation showed that siliconized HF implanted for 105 days were accepted (there was thin fibrotic layer on the external surface of the HF, no surrounding haemopoietic cells were found). It is concluded that proposed methods for preliminary evaluation of hollow fibers chosen for immunoisolation seems to be reliable and suitable for testing diffusive permeability of each relevant cell product.

Blood Flow Velocity↗

Single muscle fiber gene expression in human skeletal muscle: validation of internal control with exercise.

Reverse transcription and real-time PCR have become the method of choice for the detection of low-abundance mRNA transcripts obtained from small human muscle biopsy samples. GAPDH, beta-actin, beta-2M, and 18S rRNA are widely employed as endogenous control genes, with the assumption that their expression is unregulated and constant for given experimental conditions. The aim of this study was to determine if mRNA transcripts could be performed on isolated human single muscle fibers and to determine reliable housekeeping genes (HKGs) using quantitative gene expression protocols at rest and in response to an acute exercise bout. Muscle biopsies were obtained from the gastrocnemius of three adult males before, immediately after, and 4 h following 30 min of treadmill running at 70% of VO2max. A total of 40 single fibers (MHC I and IIa) were examined for GAPDH, beta-actin, beta-2M, and 18S rRNA using quantitative RT-PCR and SYBR Green detection. All analyzed single fiber segments showed ribosomal RNA (28S/18S). No degradation or additional bands below ribosomal were detected (rRNA ratio 1.5-1.8). Also, no high or low-molecular weight genomic DNA contamination was observed. For each housekeeping gene the duplicate average SD was +/-0.13 with a CV of 0.58%. Stable expression of GAPDH was observed at all time points for each fiber type (MHC I and IIa). Inconsistent expression of beta-actin, beta-2M, and 18S rRNA was observed during the post-exercise time points for each fiber type. These data indicate that successful extraction of high quality RNA from human single muscle fibers along with quantification of mRNA of selected genes can be performed. Furthermore, exercise does influence the expression of certain HKGs with GAPDH being the most stable.

Adaptation, Physiological↗

Chemical vapor deposition fabrication and characterization of silica-coated carbon fiber ultramicroelectrodes.

Carbon fiber disk ultramicroelectrodes (UMEs) with well-defined geometries were prepared by chemical vapor deposition techniques. Transparent silica films with thicknesses from 1 to 600 microns were deposited on the cylindrical length of 5 and 10 microns carbon fibers from a SiCl4, H2, and O2 ternary precursor system at 850-1150 degrees C or sequential deposition from Si(OEt)4 as a single source precursor at 700 degrees C followed by the SiCl4, H2, and O2 precursor system. Film thickness, film adhesion to the fiber substrate, and the overall dimensions of the silica-coated carbon fiber were studied and found to be a function of the precursor system, precursor concentrations, fiber diameter, deposition time, and fiber temperature. The silica films were found to be free of microcracks and characterized by a quality seal between the carbon fiber and the coating. As a result, the silica-coated disk UME exhibits an excellent electrochemical response without the need to use an epoxy sealant at the electrode tip. Furthermore, the deposition of hard and inert ceramic materials imparts durability to fragile carbon fibers and facilitates the handling of UMEs in microenvironments. Finally, the advantage of concentric deposition about the fibers to produce a disk UME in the center of an insulating plane was used to examine the effect of the thickness of the insulating coating on the limiting current response.

Carbon↗

Water sorption and dimensional stability of three glass fiber-reinforced composites.

PURPOSE: Water sorption and dimensional stability of three fiber-reinforced composites were studied. Two composites (Vectris, FibreKor) were resin impregnated industrially, and one composite (Stick) was polymer preimpregnated but required further manual impregnation. MATERIALS AND METHODS: Bar-shaped specimens of each material were prepared according to manufacturers' instructions. The water sorption and dimensional change of each specimen were calculated according to the change in its weight and dimension before and after immersion. Specimens were immersed in distilled water for 1, 7, 60, and 180 days. SEMs were taken to examine the quality of the fiber-matrix interface. The volume percentage of fiber content of each fiber-reinforced composite was experimentally estimated. RESULTS: A general trend of increasing water sorption for each immersion period according to the material type was: Vectris < FibreKor < Stick. There were no significant differences in dimensional change among the materials and immersion periods. CONCLUSION: The preimpregnated fiber-reinforced composite (Stick) showed higher water sorption than the industrially impregnated fiber-reinforced composites (Vectris, FibreKor). Despite a variation in the water sorption of the fiber-reinforced composites studied, all were within a 32 microg/mm3 criterion established by the ISO. The magnitude of dimensional change was small enough that it should not raise any significant clinical concern.

Absorption↗

Patterns of sensory recovery in the lingual nerve after surgical trauma.

OBJECTIVE: To examine the pattern of sensory recovery in the lingual nerve following injury, with an emphasis on qualitative changes. METHODS: Using data from one patient with prolonged dysaesthesia after surgical trauma to the lingual nerve, we correlated changes in perceived sensory function during recovery with results from qualitative and quantitative taste tests, and patient-reported quality-of-life measures. RESULTS: Recovery of quantitative taste testing as measured by automated electrogustometry, and qualitative whole of mouth taste testing, preceded perceptual recovery of taste. On the other hand, taste acuity as measured by regional qualitative taste testing tended to lag behind perceptual recovery. The pattern of recovery in the different qualitative taste sensations suggests variation in sensitivity of the nerve fibres responsible for carrying taste sensation, with the fibres relating to bitter and sour taste being the last to recover. CONCLUSION: To our knowledge, taking into account the limitations of this study, this picture of differential recovery of taste function in the lingual nerve has not been previously reported.

Electrodiagnosis↗

Removal of aqueous-phase polynuclear aromatic hydrocarbons using aspen wood fibers.

Roadway runoff derived polynuclear aromatic hydrocarbons (PAHs) impact the quality of surface and ground water. Inexpensive aspen wood fibers have been investigated as a means to remove dissolved PAH under laboratory conditions. Our isotherm experiments demonstrated that the uptake of naphthalene, fluorene, anthracene, and pyrene required up to 12.5 days to reach equilibrium. Aspen wood-water sorption coefficients, Kww, were linearly correlated to octanol-water partition coefficients and the molecular weight of the studied PAH compounds. The correlation between Kww and molecular weight was the most significant. Column experiments were carried out to study the sorption and desorption of fluorene, anthracene, and pyrene under dynamic conditions. The results indicate linear sorption, but non-linear desorption behavior. The degree of desorption was inversely correlated to a compound's hydrophobicity. Flow interruption experiments showed that sorption and desorption was rate limited. A mass balance of the sorption and desorption tests indicated that sorptive uptake exceeded desorptive release over a given number of pore volumes. Further, absolute mass-removal efficiency increased with the molecular weight and hydrophobicity of the PAH compound. Batch and column studies demonstrated that aspen wood has the potential to become an effective remedial agent for PAH in stormwater runoff or other PAH contaminated waters.

Adsorption↗

Fiber-optic probes for in vivo Raman spectroscopy in the high-wavenumber region.

In vivo Raman spectroscopy, using fiber-optic probes is hindered by the intense background signal, which is generated in the fused-silica fibers, in the fingerprint region of the Raman spectrum (approximately 0-2000 cm(-1)). Optical filtering is necessary to obtain tissue spectra of sufficient quality. The complexity of fiber-optic probes for fingerprint Raman spectroscopy, in combination with size constraints and flexibility requirements for in vivo use have been a major obstacle in the development of in vivo diagnostic tools based on Raman spectroscopy. A setup for remote Raman spectroscopic tissue characterization in the high-wavenumber region ( approximately 2400-3800 cm(-1)) is presented. It makes use of a single, unfiltered, optical fiber for guiding laser light to the sample and for collecting scattered light and guiding it back to a spectrometer. Such a simple configuration is possible because the fused-silica core and cladding of the fiber present almost no Raman background signal at these wavenumbers. Several commercially available optical fibers were tested with respect to Raman signal background, to determine their suitability for in vivo Raman spectroscopy measurements in the high-wavenumber region. Different fiber core, cladding, and coating materials were tested. Silica core-silica clad fibers, with an acrylate coating and a black nylon jacket, proved to be one of the best candidates. In vitro measurements on brain tissue of a 6-month-old pig were obtained with a remote high-wavenumber Raman setup. They illustrate the low background signal generated in the setup and the signal quality obtained with a collection time of 1 s.

Animals↗

Dietary glycemic index of human immunodeficiency virus-positive men with and without fat deposition.

This study focused on dietary glycemic index because insulin resistance can be important in the pathogenesis of fat deposition in human immunodeficiency virus (HIV). We evaluated differences in past dietary glycemic intake between men with HIV who developed fat deposition and those who did not. This was a nested case-control study consisting of 37 cases and 37 controls from the Nutrition for Healthy Living cohort. Food records from 6 to 24 months prior to development of fat deposition in cases were analyzed and compared with controls. Cases were defined as men with a waist-to-hip ratio >0.95 and body mass index (calculated as kg/m(2)) between 23 and 26. Controls were matched by age, race, body mass index, highly active antiretroviral therapy use, and CD4 count. Food records were analyzed using t tests for normally distributed nutrients and Wilcoxon rank-sum tests for nutrients with skewed distributions. Glycemic index was calculated for each meal and day. There was no significant difference in glycemic index for meals and day between participants with or without fat deposition. Both groups had a moderate dietary glycemic index intake. This study showed no association between dietary glycemic index and development of fat deposition in HIV. Instead, results of this study depict the potential benefits associated with eating high-quality diets, primarily adequate fiber and protein intake. Diet can be important in preventing development of fat deposition in patients with HIV.

Adiposity↗

[General principles of lip surgery].

A successful lip reconstruction gives a shape illusion and re-establishes physiological function. We must be able to respect static and dynamic function and the axiom "use of lip to repair lip defect". Local flaps provide optimal tissue to replace the lip in the surrounding structures, to respect aesthetic subunits and commissural function, to give a neuro-muscular continuity respecting fibers direction and mucosal quality. If extra-labial tissue is necessary, the use of the same principles enables the best compromise in shape and function as possible.

Esthetics↗

QTL mapping of seed coat color for yellow seeded Brassica napus.

The development of yellow-seeded varieties of Brassica napus for improving the oilseed quality characteristics of lower fiber content and higher protein and oil content has been a major focus of breeding researches worldwide in recent years. With the black-seeded 'Youyan 2' as male and the yellow-seeded GH06 as female parents respectively, F2 population of 132 individuals were obtained. A linkage map was constructed with 164 markers including 125 AFLP, 37 SSR, 1 RAPD and 1 SCAR markers distributed over 19 linkage groups covering approximately 2 549.8 cM with an average spacing of 15.55 cM. Two loci located on the 5th and 19th group were detected for the trait of seed coat color based on the linkage group using multiple interval mapping method and explained 46% and 30.9% of the phenotypic variation, respectively.

Amplified Fragment Length Polymorphism Analysis↗

Characteristics and use of okara, the soybean residue from soy milk production--a review.

Large quantities of okara produced annually pose a significant disposal problem. It contains mostly crude fiber composed of cellulose, hemicellulose, and lignin, about 25% protein, 10-15% oil, but little starch or simple carbohydrates. It is a suitable dietary additive in biscuits and snacks because it reduces calorie intake and increases dietary fiber. The high-quality protein fraction has good water holding and emulsifying qualities and contains a peptide with anti-hypertension effects. The pectic polysaccharides fraction is suitable for thickening acid milk products. Okara fermented with Actinomucor elegans (meitauza), Aspergillus oryzae (koji), Neurospora intermedia (ontjom), and Rhizopus oligosporus (tempe), on consumption, reduces cholesterol level and contains substances that counteract dietary free radicals. Unique and useful products produced by Bacillus subtilis and Penicillium simplicissimum on okara include surfactin and iturin A (fungicidal), okaramines A, B, D-F (D is insecticidal), an oleanane triterpene, and two dihydroquinolinones (one toxic for Artemia salina). Okara has been used as silkworm food and in the production of ceramics.

Animals↗

Transcellular water transport and stability of expression in aquaporin 1-transfected LLC-PK1 cells in the development of a portable bioartificial renal tubule device.

We investigated a portable bioartificial renal tubule device (BRTD) consisting of renal tubule cells and hollow fibers, to improve the quality of life of patients. It is necessary for a BRTD system to be compact. A compact portable BRTB requires transfection of an appropriate water channel or electrical pump genes in tubular epithelial cells, which should be based on physiological similarities to human kidney function. LLC-PK(1) cells, into which rat kidney aquaporin 1 (AQP1) cDNA was stably transfected, were evaluated for water transport ability. The expression and localization of water AQP1 were examined by Western blotting, RT-PCR, and immunofluorescence. To measure transcellular water permeation, a simple method was applied, using phenol red as a cell-impermeant marker of concentration. In contrast to wild-type LLC-PK(1) cells, rat AQP1-transfected cells had high transcellular osmotic water permeability. The expression of rat AQP1 mRNA (ratio of AQP1 to beta-actin mRNA) and protein bands (a 28-kDa band and a broad, 35- to 45-kDa band) was confirmed to be stably maintained until a population doubling level of 24. In AQP1-transfected LLCPK(1) cells, the protein was localized mainly to the basolateral side, but also the apical side, of the plasma membrane. Wild-type LLC-PK(1) cells were not stained at the plasma membrane. It is possible that enough AQP1-transfected tubule epithelial cells were supplied for a bioartificial renal tubule device.

Animals↗

Analysis of efficiency of magnetic stimulation.

Magnetic stimulation can activate excitable tissues noninvasively. However, this method requires high energy to operate and can produce equipment heat that leads to inefficient stimulation. In this study, a comprehensive optimization of efficiency for magnetic stimulation has been conducted. A total of 16 781 coil designs were tested in order to determine the optimal coil geometry and inductance for neural excitation. Induced electric fields were calculated to find the optimal stimulation site (OSS) of a given coil. The threshold energy of a magnetic pulse for neural excitation was then calculated based on the transmembrane responses of a nerve model. Simulation results show that there exists an optimal inductance, as a consequence of an optimal pulse duration, corresponding to a minimum threshold energy. A longer pulse width is required to obtain the maximum efficiency for axons with slower membrane dynamics, a longer coil-to-fiber distance, and greater values of resistance (R) and capacitance (C) of the resistance-inductance-capacitance circuit. The optimal geometry features a minimum coil height, suggesting a flat coil design for optimal efficiency. The dimension of the optimal coil design increases with the coil-to-fiber distance. Moreover, the cloverleaf design achieves the highest efficiency for infinitely long fibers whereas the butterfly design is optimal for terminating or bending fibers.

Animals↗

Exercise-limiting factors in respiratory distress.

Exercise performance data, circulatory function and respiratory and leg muscle quality, expressed as muscle fiber composition, are reviewed and together with our own data discussed as possible limiting factors for physical performance in chronic obstructive pulmonary disease (COPD). COPD is regarded as synonymous with reduced physical performance, exaggerated breathlessness or dyspnea, muscle hypotrophy and/or wasting and, frequently, malnutrition. Impaired right ventricular circulatory function seems to be essential. The observed preponderance of fast twitch (FT), 'glycogenolytic' and capillary-poor muscle fiber type in the investigated muscles might reflect endowment, a 'hypoxic vasoconstriction'-related downregulation of the other main fiber type: the slow twitch (ST), capillary-rich, fatigue-resistant fiber, and/or selective muscle trauma to ST fibers. Ischemic heart disease (IHD) patients demonstrate a similar fiber type pattern in leg muscles. Both COPD and IHD patients have low leg muscle and plasma deposits of antioxidants such as coenzyme Q10 (CoQ10) and alpha-tocopherol. This could reflect a depressed resistance to radical induced cell trauma and/or malnutrition. The magnitude of the antioxidant reduction is less pronounced in patients rich in FT fibers indicating a ST fiber-related susceptibility to trauma. Treatment of other muscle disorders including heart muscle with, e.g., CoQ10 improves performance due to a causative enhanced antioxidant potential, reduced catabolism and/or an upregulated muscle anabolism, increased mitochondrial volume/function, etc. Such data are lacking in COPD.

Aged↗

A parallel comparison of age-related peripheral nerve changes in three different strains of mice.

Strain-specific differences contributing to spontaneous age-related peripheral nerve changes were examined in three different strains of 100-week-old female mice housed under the same conditions over the same period: inbred C57BL and C3H strains, and the hybrid B6C3F1 strain. A lower incidence of obesity and significantly lower body weight, grasping power of fore- and hind-limbs, blood lipid level, tail-flick latency and motor nerve conduction velocity were observed in C57BL mice; significantly lower body temperature, blood glucose and HbA1c levels were observed in C3H mice. Histological examination conducted on isolated sciatic nerves and brachial plexuses revealed peripheral nerve lesions, characterized by axonal degeneration and remyelination, in all strains. Although the extent of histopathologic change in nerve fibers was similar in quality to those observed in all three mouse strains, the incidence and severity of nerve lesions in B6C3F1 and C3H mice were significantly greater than those observed in C57BL mice.

Aging↗

An improved method for embedding brain tissue in albumin-gelatin.

A new modification of the Snodgrass-Dorsey (1963) albumin embedding method is described. Formalin fixed brains of various ages of rhesus monkey (Macaca mulatta) were sunk in 10% phosphate buffered formalin which contained 30% sucrose, and then embedded in a 3% gelatin, 30% egg albumin solution which had been centrifuged to ensure uniformity. The albumin-gelatin was hardened in formaldehyde fumes and blocks cut frozen at 10-40 micron. Sections thus prepared can be handled easily and mounted without damage to the tissue. Modifications of conventional cell and fiber stains produce high quality finished slides in which the stained brain tissue is surrounded by a colorless albumin-gelatin matrix.

Animals↗

Whole-herd optimization with the Cornell Net Carbohydrate and Protein System. III. Application of an optimization model to evaluate alternatives to reduce nitrogen and phosphorus mass balance.

The objectives of this paper were to use a linear programming model previously described to evaluate different alternatives for reducing excess nutrients that may influence water quality on a case study farm (300 lactating cows on 430 ha of cropland growing alfalfa, grass, and corn). Several alternatives perceived to influence farm nutrient balance were evaluated for their potential to reduce N and P mass balance. Dividing lactating cow diets into three groups according to their level of milk production versus a one-group total mixed ration decreased mass balance (tonne/yr) from 51.7 to 44.7 for N, from 6.7 to 6.1 for P and from 16.2 to 14.8 for K with little influence on return over feed costs. Increasing forage quality (lower neutral detergent fiber and higher crude protein) did not improve N balance because of the increased N fixation from the air to the soil, but it increased returns over feed costs by $31,385. Improving yields to the maximum potential for the farm reduced mass balance by 29, 51, and 100% for N, P, and K, respectively, and increased returns over feed costs by $70,579. Changing the crop hectare proportions to more corn and less alfalfa reduced N and K balances by 19 and 29%, respectively, and increased returns over feed costs $39,383. Increasing annual milk production 10% by increasing milk production per head 10% compared with increasing animal numbers at the current average milk production per cow until total milk increased 10% gave $34,132 more return over feed costs with less N, P, and K retained on the farm.

Agriculture↗

Performance of an in-vivo, continuous blood-gas monitor with disposable probe.

A fluorescence-based fiber optic measurement system has been developed for monitoring pH, pCO2, and pO2 through a 20-gauge radial artery catheter without compromising capabilities for monitoring arterial pressure or for blood withdrawal. The measuring probe consists of three optical fibers to which the sensing chemistries are attached, and a thermocouple that measures temperature. The probe is designed to meet in-vivo biocompatibility requirements for a one-time use of up to 72 h. The components that are in contact with the patient's blood are nontoxic, nonhemolytic, nonthrombogenic, and sterilizable. Blood compatibility is enhanced by including covalently bound heparin. The in-vitro accuracy of the system has been tested against commercial blood-gas measurement instruments; comparison with tonometry and blood gas values gave r greater than or equal to 0.98 for all three sensors. The standard error for all sensors was within the College of American Pathologists' accuracy guidelines for measuring blood gas. Instrumental drift was minimal, indicating that system performance characteristics should not be the limiting factor in obtaining clinically useful information for up to 72 h. The response times of the sensors in animal and in-vitro studies were less than 2 min, suitable for monitoring physiological changes in blood gas values.

Animals↗