Viscometric and static light scattering studies on an exopolysaccharide produced by Lactobacillus delbrueckii subspecies bulgaricus NCFB 2483

The rheological properties and molecular parameters of a purified exopolysaccharide (EPS) produced by a ropy strain of Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483 were investigated. Using capillary viscometry, an intrinsic viscosity of 2,013 mL/g was obtained. The flow curves were fitted by both the Carreau and the Cross equations for shear-thinning fluids, with the Carreau equation giving a better fit. The Cross equation fitted fairly well the plot of reduced viscosity as a function of reduced shear rate with an exponent value (1 - n) of approximately 0.76, typical of random coil polymers. Furthermore, the concentration dependence of the viscosity plot showed a gradient of approximately 1.1 in the dilute regime and 3.3 in the semidilute regime. Molecular parameters were obtained using a multiangle laser light scattering technique. The 2483 EPS molecules had a weight-average molar mass of approximately 2 x 10(6) Da and a z-average root mean square radius (RMS) of approximately 151 nm. From the light scattering data, the bacterial EPS was also found to have a low polydispersity index (approximately 1.15) and adopt a random coil conformation.     

Periodic fluid extrusion and models of digesta mixing in the intestine of a herbivore,the common brushtail possum (<Emphasis Type="Italic">Trichosurus vulpecula</Emphasis>)

The digesta in four gut compartments (proximal and distal halves of small intestine, caecum, and proximal colon) of a wild hindgut fermenting herbivore, the common brushtail possum (Trichosurus vulpecula), were investigated by rheometry and permeametry. Digesta from all compartments were highly viscous and exhibited shear-thinning. Apparent viscosity was positively related to dry matter content, and increased from proximal small intestine to colon. Dynamic rheological measurements showed that in small intestinal digesta the elastic modulus was greater than the viscous modulus and their ratios were characteristic of weak gels, indicating that digesta could sustain compression. The apparent viscosity of distal small intestinal digesta was markedly lower when measured by capillary viscometry than by rotatory viscometry, indicating that plug flow was likely to be facilitated by lubrication from a peripheral layer of less viscous fluid; i.e., there was an augmented plug flow. Permeametry showed that fluid was extruded from all digesta on compression at physiological pressures, that there was significant permeability of proximal and distal small intestinal digesta, but that digesta became progressively compacted during permeation, with a concomitant reduction in permeability as dry matter content increased. It is proposed that conditions within the small intestine differ from those of an ideal plug flow reactor as radial mixing and turbulence cannot occur. Instead, we suggest that segmentation and peristalsis aid radial mixing of the fluid phase by compressing the solid phase, with extrusion of fluid through the digesta plug. This extrusion may be followed by resorption of fluid back into the plug when the elasticity of the solid phase of digesta is Hookean, thus aiding the mixing of secreted enzymes with insoluble substrates within the plug.     

Assessing the in vitro fitness of an oseltamivir-resistant seasonal A/H1N1 influenza strain using a mathematical model

In 2007, the A/Brisbane/59/2007 (H1N1) seasonal influenza virus strain acquired the oseltamivir-resistance mutation H275Y in its neuraminidase (NA) gene. Although previous studies had demonstrated that this mutation impaired the replication capacity of the influenza virus in vitro and in vivo, the A/Brisbane/59/2007 H275Y oseltamivir-resistant mutant completely out-competed the wild-type (WT) strain and was, in the 2008-2009 influenza season, the primary A/H1N1 circulating strain. Using a combination of plaque and viral yield assays, and a simple mathematical model, approximate values were extracted for two basic viral kinetics parameters of the in vitro infection. In the ST6GalI-MDCK cell line, the latent infection period (i.e., the time for a newly infected cell to start releasing virions) was found to be 1-3 h for the WT strain and more than 7 h for the H275Y mutant. The infecting time (i.e., the time for a single infectious cell to cause the infection of another one) was between 30 and 80 min for the WT, and less than 5 min for the H275Y mutant. Single-cycle viral yield experiments have provided qualitative confirmation of these findings. These results, though preliminary, suggest that the increased fitness success of the A/Brisbane/59/2007 H275Y mutant may be due to increased infectivity compensating for an impaired or delayed viral release, and are consistent with recent evidence for the mechanistic origins of fitness reduction and recovery in NA expression. The method applied here can reconcile seemingly contradictory results from the plaque and yield assays as two complementary views of replication kinetics, with both required to fully capture a strain's fitness.     

TNF deficiency fails to protect BAFF transgenic mice against autoimmunity and reveals a predisposition to B cell lymphoma

TNF is well characterized as a mediator of inflammatory responses. TNF also facilitates organization of secondary lymphoid organs, particularly B cell follicles and germinal centers, a hallmark of T-dependent Ab responses. TNF also mediates defense against tumors. We examined the role of TNF in the development of inflammatory autoimmune disorders resembling systemic lupus erythematosus and Sj?gren's syndrome induced by excess B cell-activating factor belonging to the TNF family (BAFF), by generating BAFF-transgenic (Tg) mice lacking TNF. TNF(-/-) BAFF-Tg mice resembled TNF(-/-) mice, in that they lacked B cell follicles, follicular dendritic cells, and germinal centers, and have impaired responses to T-dependent Ags. Nevertheless, TNF(-/-) BAFF-Tg mice developed autoimmune disorders similar to that of BAFF-Tg mice. Disease in TNF(-/-) BAFF-Tg mice correlates with the expansion of transitional type 2 and marginal zone B cell populations and enhanced T-independent immune responses. TNF deficiency in BAFF-Tg mice also led to a surprisingly high incidence of B cell lymphomas (>35%), which most likely resulted from the combined effects of BAFF promotion of neoplastic B cell survival, coupled with lack of protective antitumor defense by TNF. Thus, TNF appears to be dispensable for BAFF-mediated autoimmune disorders and may, in fact, counter any proneoplastic effects of high levels of BAFF in diseases such as Sj?gren's syndrome, systemic lupus erythematosus, and rheumatoid arthritis.     

Regional distribution of thermal sensitivity to cold at rest and during mild exercise in males

Although several studies have compared thermal sensitivity between body segments, little is known on regional variations within body segments. Furthermore, the effects of exercise on the thermal sensation resulting from a cold stimulus remain unclear. The current experiment therefore aimed to explore inter- and intra-segmental differences in thermal sensitivity to cold, at rest and during light exercise. Fourteen male participants (22.3±3.1 years; 181.6±6.2 cm; 73.7±10.3 kg) were tested at rest and whilst cycling at 30% VO_2 max. Sixteen body sites (front torso=6; back=6; arms=4) were stimulated in a balanced order, using a 20 °C thermal probe (25 cm²) applied onto the skin. Thermal sensations resulting from the stimuli were assessed using an 11-point cold sensation scale (0=not cold; 10=extremely cold). Variations were found within body segments, particularly at the abdomen and mid-back where the lateral regions were significantly more sensitive than the medial areas. Furthermore, thermal sensations were significantly colder at rest compared to exercise in 12 of the 16 body sites tested. Neural and hormonal factors were considered as potential mechanisms behind this reduction in thermal sensitivity. Interestingly, the distribution of cold sensations was more homogenous during exercise. The present data provides evidence that thermal sensitivity to cold varies within body segments, and it is significantly reduced in most areas during exercise.