Weekly self-monitoring and treatment adjustment benefit patients with partly controlled and uncontrolled asthma: an analysis of the SMASHING study

Background

Internet-based self-management has shown to improve asthma control and asthma related quality of life, but the improvements were only marginally clinically relevant for the group as a whole. We hypothesized that self-management guided by weekly monitoring of asthma control tailors pharmacological therapy to individual needs and improves asthma control for patients with partly controlled or uncontrolled asthma.

Methods

In a 1-year randomised controlled trial involving 200 adults (18-50 years) with mild to moderate persistent asthma we evaluated the adherence with weekly monitoring and effect on asthma control and pharmacological treatment of a self-management algorithm based on the Asthma Control Questionnaire (ACQ). Participants were assigned either to the Internet group (n = 101) that monitored asthma control weekly with the ACQ on the Internet and adjusted treatment using a self-management algorithm supervised by an asthma nurse specialist or to the usual care group (UC) (n = 99). We analysed 3 subgroups: patients with well controlled (ACQ ≤ 0.75), partly controlled (0.75>ACQ ≤ 1.5) or uncontrolled (ACQ>1.5) asthma at baseline.

Results

Overall monitoring adherence was 67% (95% CI, 60% to 74%). Improvements in ACQ score after 12 months were -0.14 (p = 0.23), -0.52 (p < 0.001) and -0.82 (p < 0.001) in the Internet group compared to usual care for patients with well, partly and uncontrolled asthma at baseline, respectively. Daily inhaled corticosteroid dose significantly increased in the Internet group compared to usual care in the first 3 months in patients with uncontrolled asthma (+278 μg, p = 0.001), but not in patients with well or partly controlled asthma. After one year there were no differences in daily inhaled corticosteroid use or long-acting β₂-agonists between the Internet group and usual care.

Conclusions

Weekly self-monitoring and subsequent treatment adjustment leads to improved asthma control in patients with partly and uncontrolled asthma at baseline and tailors asthma medication to individual patients'' needs.

Trial registration

Current Controlled Trials ISRCTN79864465     

Engineering the cell surface display of cohesins for assembly of cellulosome-inspired enzyme complexes on <Emphasis Type="Italic">Lactococcus lactis</Emphasis>

Background  

The assembly and spatial organization of enzymes in naturally occurring multi-protein complexes is of paramount importance for the efficient degradation of complex polymers and biosynthesis of valuable products. The degradation of cellulose into fermentable sugars by Clostridium thermocellum is achieved by means of a multi-protein "cellulosome" complex. Assembled via dockerin-cohesin interactions, the cellulosome is associated with the cell surface during cellulose hydrolysis, forming ternary cellulose-enzyme-microbe complexes for enhanced activity and synergy. The assembly of recombinant cell surface displayed cellulosome-inspired complexes in surrogate microbes is highly desirable. The model organism Lactococcus lactis is of particular interest as it has been metabolically engineered to produce a variety of commodity chemicals including lactic acid and bioactive compounds, and can efficiently secrete an array of recombinant proteins and enzymes of varying sizes.     

Steroid structural requirements for stabilizing or disrupting lipid domains

In artificial membrane bilayers, saturated long acyl chain-containing phospholipids and cholesterol (Chol) interact to form more ordered domains than those in phospholipids with unsaturated or short fatty acyl chains. We have extended the fluorescence techniques of London et al. [Xu, X., and London, E. (2000) Biochemistry 39, 843-849; Xu, X., Bittman, R., Duportail, G., Heissler, D., Vilchezes, C., and London, E. (2001) J. Biol. Chem. 276, 33540-33546] to study the propensity of several steroids to form or disrupt such ordered lipid domains. Temperature-dependent fluorescence quenching and steady-state polarization of the extrinsic fluorescent probe diphenylhexatriene (DPH) in model membranes composed of dipalmitoylphosphatidylcholine (or sphingomyelin), a nitroxide spin-labeled phosphatidylcholine (12-SLPC), and a given steroid were combined to study the influence of the latter on (a) ordered lipid domain formation, (b) stabilization, and (c) the extension of the ordered lipid assemblies. The results of the two totally independent methods, fluorescence quenching by 12-SLPC and fluorescence polarization of DPH, show that all steroids examined, except for Chol and 25-hydroycholesterol, behave as lipid domain-disrupting compounds. Additionally, we found a positive correlation between the hydrophobicity of steroids and their ordered lipid domain-promoting activity. Comparison of the chemical structures disclosed some distinctive traits of ordered lipid domain-promoting steroids: (i) the presence of an isooctyl side chain bond at C17; (ii) the absence of carbons attached to C23 (i.e., C24-C27) in any of the other (domain-disrupting) steroids; (iii) the presence of a small polar group at position C3; and (iv) the absence of polar groups in the fused rings, with the exception of substitutions at position C3 in the A ring.     

The AUG start codon of the Saccharomyces cerevisiae NFS1 gene can be substituted for by UUG without increased initiation of translation at downstream codons.

The selection of the site for initiation of translation for the Saccharomyces cerevisiae NFS1 gene was examined using mutated AUG1, AUG2 and AUG3 codons. When AUG1 of the yeast NFS1 gene was mutated to UUG and the resulting mRNA was translated in vitro using a reticulocyte system, initiation from the mutated codon was abolished and occurred instead at downstream codons at increased rates. When the same mRNA was translated using a yeast extract, translation initiated at the mutated codon, albeit at a reduced rate, and there was no increased translation at downstream AUG codons. The NFS1 gene in which AUG1 was replaced by UUG was also able to substitute for the wild-type gene in vivo in yeast. Western blots confirmed that the encoded protein was the same size as that encoded by the wild-type gene and that both the wild-type and mutated proteins localized to mitochondria. This is apparently the first example of a yeast protein where mutagenesis of AUG1 does not lead to alternate use of a downstream AUG.     

The abundance of additional N-acetyllactosamine units in N-linked tetraantennary oligosaccharides of human Tamm-Horsfall glycoprotein is a donor-specific feature

Previously, treatment of Tamm-Horsfall glycoprotein (THp) from different donors with endo-beta-galactosidase has been shown to liberate a tetra- and a Sd(a)-active pentasaccharide, concluding the presence of N-linked carbohydrate chains containing additional N - acetyllactosamine units. These type of oligosaccharides were not found in a detailed structure elucidation of the carbohydrate moiety of THp of one male donor, suggesting a donor-specific feature for these type of structures. Therefore, THp was isolated from four healthy male donors and each subjected to endo-beta-galactosidase treatment in order to release these tetra- and Sd(a)-active pentasaccharide. Differences were observed in the total amount of released tetra- and Sda-active pentasaccharide of the used donors (42, 470, 478, 718 microg/100 mg THp), indicating that the presence of repeating N-acetyllactosamine units incorporated into the N-glycan moiety of THp is donor specific. Furthermore, a higher expression of the Sd(a) determinant on antennae which display N-acetyllactosamine elongation was observed, suggesting a better accessibility for the beta-N-acetylgalactosaminyltransferase. In order to characterize the N-glycans containing repeating N- acetyllactosamine units, carbohydrate chains were enzymatically released from THp and isolated. The tetraantennary fraction, which accounts for more than 33% of the total carbohydrate moiety of THp, was used to isolate oligosaccharides containing additional N - acetyllactosamine units. Five N-linked tetraantennary oligosaccharides containing a repeating N-acetyllactosamine unit were identified, varying from structures bearing four Sd(a) determinants to structures containing no Sd(a) determinant (see below). One compound was used in order to specify the branch location of the additional N- acetyllactosamine unit, and it appeared that only the Gal-6' and Gal-8' residues were occupied by a repeating N -acetyllactosamine unit.      

The connection between ionophore-mediated Ca2+-movements and intermediary metabolism in human red cells. I. Relationships between Ca2+-loading,ATP-consumption and glycolytic flux

Human red cells (RBC) were loaded with moderate amounts of Ca²⁺ by the ionophore A23187. Quantitative relationships between Ca²⁺-loading, ATP consumption and glycolytic flux were established. 1. Ca²⁺-loading is accompanied by ATP depletion. A maximum ATP consumption of approximately 10 mmoles/l RBC/h was estimated. 2. There is a positive correlation between lactate formation and Ca²⁺-loading. This is linear from 1.4 to about 4 mmoles lactate/l RBC/h. 3. Ca²⁺-induced glycolytic stimulation seems not to be mediated by adenine nucleotides. A wide range of energy charges and very different adenine nucleotide patterns were associated with the same stimulation of lactate production. 4. The turnover of the (Ca²⁺-Mg²⁺)-ATPase and its share in the Ca²⁺-stimulated ATP consuming processes were estimated with inhibitors. 1 mM La³⁺ inhibited both Ca²⁺-outward transport and ATP consumption by 80%. The remaining 20% of the ATP consumption was accounted for by the (Na⁺-K⁺)-ATPase. 5. A Ca²⁺ extrusion to ATP consumption molar ratio of 2:1 was found. However, when ATP consumption was due to the breakdown of previously accumulated glycolytic intermediates, the ratio dropped to about 1.