The effect of butanol challenge (0, 1.0, 1.5% [vol/vol]) and growth temperature (22, 37, 42°C) on the membrane composition and fluidity of Clostridium acetobutylicum ATCC 824 and a butanol-tolerant mutant, SA-2, was examined in chemically defined medium. Growth of strain ATCC 824 into the stationary phase coincided with a gradual increase in the percent saturated to percent unsaturated (SU) fatty acid ratio. When challenged with butanol at 22 and 37°C, ATCC 824 demonstrated an immediate (within 30 min) dose-response increase in the SU ratio. This strain showed little additional change over a 48-h fermentation. Compared with ATCC 824, growth of SA-2 into the late stationary phase at 22 or 37°C resulted in an overall greater increase in the SU ratio for both unchallenged and challenged cells. This effect was minimized when SA-2 was challenged at 42°C, probably due to the combination of the membrane fluidizing effect of butanol and the elevated temperature. Growth at 42°C resulted in an increase in longer acyl chain fatty acids at the expense of shorter acyl chains for both strains. The membrane fluidity exhibited by SA-2 remained essentially constant at various butanol challenge and temperature combinations, while that for the ATCC 824 strain increased with increasing butanol challenge. By synthesizing an increased amount of saturated fatty acids, the butanol-tolerant SA-2 strain has apparently developed a mechanism for maintaining a more stable membrane environment. Growth of the microorganism is necessary for butanol to fluidize the membrane. Incorporation of exogenous fatty acids (18:1) did not significantly improve the butanol tolerance of either strain. Since SA-2 was able to produce only trace amounts of either butanol or acetone, increased tolerance to butanol does not necessarily coincide with greater solvent yields in this strain.

Cardiometabolic impairments that begin early in life are particularly critical, as they often predict metabolic dysfunction into adulthood. Obesity, high-fat diet (HFD), and inactivity are all associated with adipose tissue (AT) inflammation and insulin resistance (IR), major predictors of metabolic dysfunction. Recent evidence also has associated the gut microbiome with cardiometabolic health. PURPOSE Compare equal energy deficits induced by exercise and caloric reduction on cardiometabolic disease risk parameters including AT inflammation, IR and gut microbiota changes during HFD consumption. METHODS Obesity-prone rats fed HFD were exercise trained (Ex, n=10) or weight-matched to Ex via caloric reduction while kept sedentary (WM, n=10), and compared to ad libitum HFD-fed (Sed, n=10) rats for IR, systemic energetics and spontaneous physical activity (SPA), adiposity, and fasting metabolic parameters. Visceral, subcutaneous, periaortic, and brown AT, liver, aorta, and cecal digesta were examined. RESULTS Despite identical reductions in adiposity, Ex, but not WM, improved IR, increased SPA by ~26% (p<0.05 compared to WM and Sed), and reduced LDL cholesterol (p<0.05 compared to Sed). WM and Ex both reduced inflammatory markers in all AT depots and aorta, while only Ex increased indicators of mitochondrial function in brown AT. Ex significantly increased the relative abundance of cecal Streptococcaceae and decreased S24-7 and one undefined genus in Rikenellacea; WM induced similar changes that did not reach statistical significance. CONCLUSIONS Both Ex and WM reduced AT inflammation across depots, while Ex caused more robust changes to gut microbial communities, improved IR, increased fat oxidation, increased SPA, and increased indices of brown AT mitochondrial function. Our findings add to the growing body of literature indicating that there are weight-loss independent metabolic benefits of exercise.

This study documents the discrete solute permeability mechanisms associated with physiologically high concentrations of human alpha-thrombin and bradykinin stimulation of bovine pulmonary artery endothelial cell (BPAEC) monolayers using fluorescein isothiocyanate-hydroxyethyl starch macromolecules. Agonist-induced alterations of intracellular free calcium ([Ca2+]i) using fura-2 acetoxymethyl ester were also measured. BPAEC monolayers showed restricted diffusion consistent with a small-pore (approximately 150 A) radius under baseline conditions. Thrombin produced a major increase in monolayer permeability that was greatest for solute molecular radii (ae) > 100 A. This effect was associated with the exposure of the large (approximately 2,000 A) pores of the filter support by 50- to 1,050-microns2 open areas between approximately 0.5% of the adjacent endothelial cells. This heterogeneous endothelial barrier of parallel large- and small-pore transport pathways permitted solute convection with free diffusion across a few large pores to dominate the restricted diffusion of most apparently unperturbed endothelial junctions. Bradykinin produced a small, transient elevation in monolayer permeability to ae < 35 A, consistent with an increase in the number of small pores or a decrease in path length of this transport pathway. The bradykinin- and thrombin-induced peak elevations in [Ca2+]i were inversely associated with the degree of increased monolayer solute permeability, and enzymatically inhibited thrombin produced none of these effects. These data show that bradykinin and human alpha-thrombin represent two distinct classes of endothelial cell agonists that initiate discrete solute permeability mechanisms.