In vitro fermentation characteristics of selected glucose-based polymers by canine and human fecal bacteria

This research evaluated fermentation characteristics (short-chain fatty acid [SCFA] production, pH, and gas production) resulting from fermentation of glucose-based carbohydrates using canine (n = 3) and human (n = 3) fecal inoculum. Substrates included lyophilized canine ileal digesta containing maltodextrin, gamma-cyclodextrin, high molecular weight (MW) pullulan (MW 100000), or low MW pullulan (MW 6300) obtained from an in vivo experiment. Fermentation for 6 and 10 h with human fecal microflora resulted in higher gas and SCFA production than did canine fecal microflora. High MW pullulan fermentation resulted in the highest (p < 0.05) gas production and lowest (p < 0.05) pH for both dogs and humans. Total SCFA production was highest (p < 0.05) for low MW pullulan fermented by canine microflora, and for gamma-cyclodextrin, high MW pullulan, and low MW pullulan fermented by human microflora. Differences were noted in fermentation characteristics of substrates present in ileal digesta.     

Combined effect of follicle-follicle interactions and declining follicle-stimulating hormone on murine follicle health in vitro

Follicle selection occurs throughout an adult female's reproductive life, with selected, dominant follicle(s) developing to the preovulatory stage whereas the remaining, subordinate follicles within the growing cohort instead undergo atresia and die. To date, most research into follicle dominance has concentrated on its endocrine regulation, although it seems likely that intraovarian mechanisms are also involved in its regulation. We demonstrate here that the response of singly cultured murine follicles to declining concentrations of FSH depends on their developmental stage, with follicles at an earlier stage of development being much more susceptible than mature follicles to a lowering of FSH levels. We then extrapolate this information to follicle cocultures, in which a large dominant follicle was grown with a small subordinate follicle in a manner that maintained a dominant/subordinate relationship, with follicle health assessed by a terminal transferase-mediated 2'-deoxyuracil 5'-triphosphate nick end-labeled reaction on whole-follicle mounts. Our investigations show a combined negative effect of coculture and FSH withdrawal on small subordinate follicles, such that subordinate follicles cocultured with dominant follicles and subjected to a lowering of FSH levels during the culture period exhibit a greatly increased incidence of apoptosis in the granulosa cells (750% increase) compared with that exhibited by the dominant follicles (97% increase). We suggest that a similar interaction between endocrine and intraovarian factors regulates follicular dominance in vivo, such that dominant follicles, in addition to bringing about a fall in FSH levels via the hypothalamic-pituitary axis, exert local, direct effects on subordinate follicles, with both of these influences combining to induce atresia in subordinate follicles.     

Exposure of yeast cells to anoxia induces transient oxidative stress. Implications for the induction of hypoxic genes

The mitochondrial respiratory chain is required for the induction of some yeast hypoxic nuclear genes. Because the respiratory chain produces reactive oxygen species (ROS), which can mediate intracellular signal cascades, we addressed the possibility that ROS are involved in hypoxic gene induction. Recent studies with mammalian cells have produced conflicting results concerning this question. These studies have relied almost exclusively on fluorescent dyes to measure ROS levels. Insofar as ROS are very reactive and inherently unstable, a more reliable method for measuring changes in their intracellular levels is to measure their damage (e.g. the accumulation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in DNA, and oxidative protein carbonylation) or to measure the expression of an oxidative stress-induced gene, e.g. SOD1. Here we used these approaches as well as a fluorescent dye, carboxy-H(2)-dichloro-dihydrofluorescein diacetate (carboxy-H(2)-DCFDA), to determine whether ROS levels change in yeast cells exposed to anoxia. These studies reveal that the level of mitochondrial and cytosolic protein carbonylation, the level of 8-OH-dG in mitochondrial and nuclear DNA, and the expression of SOD1 all increase transiently during a shift to anoxia. These studies also reveal that carboxy-H(2)-DCFDA is an unreliable reporter of ROS levels in yeast cells shifted to anoxia. By using two-dimensional electrophoresis and mass spectrometry (matrix-assisted laser desorption ionization time-of-flight), we have found that specific proteins become carbonylated during a shift to anoxia and that some of these proteins are the same proteins that become carbonylated during peroxidative stress. The mitochondrial respiratory chain is responsible for much of this carbonylation. Together, these findings indicate that yeast cells exposed to anoxia experience transient oxidative stress and raise the possibility that this initiates the induction of hypoxic genes.     

The potential influence of the heel counter on internal stress during static standing: a combined finite element and positional MRI investigation

Confinement of the heel due to the counter of the shoe is believed to influence heel pad biomechanics. Using a two-dimensional finite element model of the heel pad and shoe during a simulation of static standing, the aim of this study was to quantify the potential effect of confinement on internal heel pad stress. Non-weightbearing MRI and weightbearing MRI with plantar pressure and ground reaction force data were recorded for a single subject. The non-weightbearing MRI was used to create two FE models of the heel pad, using either homogeneous or composite material properties. The composite model included a distinction in material properties between fat pad and skin. Vertical and medial-lateral forces, as measured on the subject's heel, were applied to the models and vertical compressive strains for both models were comparable with those observed by weightbearing MRI. However, only for the composite model was the predicted plantar pressure distribution comparable with measured data. The composite model was therefore used in further analyses. In this composite model, the internal stresses were located mainly in the skin and were predominantly tensile in nature, whereas the stress state in the fat pad approached hydrostatic conditions. A representation of a running shoe, including an insole, midsole and heel counter was then added to the composite heel pad to form the shod model. In order to investigate the counter effect, the load was applied to the shod model with and without the heel counter. The effect of the counter on peak stress was to elevate compression (0-50%), reduce tension (22-34%) and reduce shear (22-28%) in the skin. In addition, the counter reduced both compressive (20-40%) and shear (58-80%) stress in the fat pad and tension in the fat pad remained negligible. Taken together the results indicate that a well-fitted counter works in sympathy with the internal structure of the heel pad and could be an effective reducer of heel pad stress. However, further research needs to be undertaken to assess the long-term effects on the soft-tissues, practicalities of achieving good fit and behavior under dynamic events.     

Preferential vastus medialis oblique activation achieved as a treatment for knee disorders

The purpose of this study was to compare the effect of an open-stance cycling protocol (OSCP) with the traditional cycling foot position (TCFP) for preferential vastus medialis oblique (VMO) muscle activation, measured by surface electromyography (SEMG), and preferential VMO activation as defined by achieving significantly increased VMO/VL (vastus lateralis muscle) ratio values. Forty subjects of both sexes participated, 18 symptomatic with patellofemoral pain and 22 control subjects; ages ranged from 18 to 60 years (mean = 28.7 +/- 8 years). The OSCP and TCFP were ridden in randomized order while SEMG recordings were taken of the VMO and VL muscles, collecting the mean of peak amplitudes to calculate VMO/VL ratio values. The SEMG readings were taken 4 times per testing session with randomized resistance and a consistent cycling cadence of 85 rpm. The OSCP displayed preferential VMO activation for all subject groups (F = 40.47, p = 0.0001), and this study revealed a protocol that effectively treats patellofemoral pain.     

Effect of prism orientation and loading direction on contact stresses in prismatic enamel of primates: implications for interpreting wear patterns

The ability of prisms to effectively dissipate contact stress at the surface will influence wear rates in teeth. The aim of this investigation was to begin to quantify the effect of prism orientation on surface stresses. Seven finite element models of enamel microstructure were created, each model differing in the angulation of prism orientation with regard to the wear surface. For validation purposes, the mechanical behavior of the model was compared with published experimental data. In order to test the enamel under lateral loads, a compressed food particle was dragged across the surface from the dentino-enamel junction (DEJ) towards the outer enamel surface (OES). Under these conditions, tensile stresses in the enamel model increased with increases in the coefficient of friction. More importantly, stresses were found to be lowest in models in which the prisms approach the surface at lower angles (i.e., more obliquely cut prisms), and highest when the prisms approached the surface at 60 degrees (i.e., less obliquely cut). Finally, the direction of travel of the simulated food particle was reversed, allowing comparison of the difference in behavior between trailing and leading edge enamels (i.e., when the food particle was dragged either towards or away from the DEJ). Stresses at the trailing edge were usually lower than stresses at the leading edge. Taken together with what is known about prism orientation in primate teeth, such findings imply greater wear resistance at the intercuspal region and less wear resistance at the lateral enamel at midcrown. Such findings appear to be supported by archeological evidence.     

Photochemistry of dyes and fluorochromes used in biology and medicine: some physicochemical background and current applications

An overview of the basic principles of photochemistry is presented to facilitate discussion of fluorescence, quenching and quantum yields. These topics in turn provide the foundation for an account of fluorescence spectroscopy and its application to microscopy. A brief overview of light microscopy and the application of fluorescence microscopy is given. The influences of molecular features, such as aromatic character and substitution patterns, on color and fluorescence are described. The concept of color fading is considered with particular reference to its effect on microscopic preparations. A survey of representative fluorescent probes is provided, and their sensitivity, application, and limitations are described. The phototoxicity of fluorescent molecules is discussed using biomembranes and DNA as examples of targets of toxicity. Photodynamic therapy, a relatively new clinical application of phototoxicity, is described. Both anticancer and antimicrobial applications are noted, and an assessment is given of the current ideas on the ideal physicochemical properties of the sensitizing agents for such applications.     

Synthesis and antinephritic activities of quinoline-3-carboxamides and related compounds.

A series of linomide-related quinoline-3-carboxamides and their analogues was prepared and evaluated for antinephritic activities. The 6-MeS derivative 7a was highly effective in two nephritis models, namely chronic graft-versus-host disease and autoimmune MRL/l mice.