Long-term analysis of diesel fuel consumption in a co-culture of Acinetobacter venetianus,Pseudomonas putida and Alcaligenes faecalis

The dynamics of a microbial population isolated from superficial waters of Venice Lagoon and the ability to utilise diesel fuel (n-alkanes mixture C₁₂-C₂₈) as the sole carbon and energy source were studied in a long-term reconstruction experiment. The reconstructed microbial population consisted of three bacterial strains belonging to the species Acinetobacter venetianus, Pseudomonas putida, and Alcaligenes faecalis, which were able to oxidise n-alkanes to alkanoates, n-alkanols to alkanoates, or only n-alkanoates, respectively. Three different approaches: plate counting, cell counting by epifluorescence microscopy with DAPI staining, and by fluorescence in situ hybridisation (FISH) by using a probe conjugate with fluoresceine isothiocyanate specifically targeted towards the 16S rRNA of bacteria belonging to the genus Acinetobacter were used to monitor the growth of the bacterial population. The growth of A. venetianus was stimulated by the presence of other strains, suggesting a beneficial interaction. After the first week of growth A. venetianus cells formed aggregates, as confirmed by confocal microscopy (CLSM), which allowed them to be distinguished from free cells. A relationship between cell number and measured areas (μm²) per aggregate was found. Each cell presented an average surface of 1.21 μm². Each aggregate was formed by a cellular monolayer biofilm consisting of up to several thousands of cells. The A. venetianus aggregates increased in number and size over time, but after two weeks fragmentation events, which had a beneficial effect on the growth of P. putida and A. faecalis, occurred. This revised version was published online in June 2006 with corrections to the Cover Date.     

Prevalence of HSP47 antigen and autoantibodies to HSP47 in the sera of patients with mixed connective tissue disease

The 47-kDa heat shock protein (HSP47) is an endoplasmic reticulum molecular chaperone that assists in the maturation of collagen molecules and whose expression is known to be upregulated in lesions of fibrotic diseases. We examined the levels of HSP47 protein and autoantibodies to HSP47 in the sera of patients with rheumatic autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, Sj?gren's syndrome, and mixed connective tissue disease (MCTD) by enzyme-linked immunosorbent assay and immunoblot analysis. Patients with idiopathic pulmonary fibrosis (IPF) were assessed as an example of non-autoimmune fibrotic disease. HSP47 antigen and autoantibody levels are significantly elevated in the sera of the rheumatic autoimmune disease patients, but not in the sera of the IPF patients. The sera of the MCTD patients showed particularly high levels of HSP47 antigen relative to healthy controls (1.99+/-0.22 vs 0.41+/-0.07 ng/ml). Autoantibodies to HSP47 were also in high levels in the sera of MCTD patients. These results suggest that simultaneous occurrence of systemic inflammation and upregulation of HSP47 caused leakage of HSP47 from fibrotic lesions into the peripheral blood, and the leaked antigen induced high titer of autoantibodies to HSP47. The high levels of HSP47 antigen and autoantibody may be useful blood markers of MCTD.     

Fisher Lecture: the 2002 R. A. Fisher lecture: dedicated to the memory of Shanti S. Gupta. Variances are not always nuisance parameters

In classical problems, e.g., comparing two populations, fitting a regression surface, etc., variability is a nuisance parameter. The term "nuisance parameter" is meant here in both the technical and the practical sense. However, there are many instances where understanding the structure of variability is just as central as understanding the mean structure. The purpose of this article is to review a few of these problems. I focus in particular on two issues: (a) the determination of the validity of an assay; and (b) the issue of the power for detecting health effects from nutrient intakes when the latter are measured by food frequency questionnaires. I will also briefly mention the problems of variance structure in generalized linear mixed models, robust parameter design in quality technology, and the signal in microarrays. In these and other problems, treating variance structure as a nuisance instead of a central part of the modeling effort not only leads to inefficient estimation of means, but also to misleading conclusions.     

Adsorption of actin at the air-water interface: a monolayer study

The intrinsic surface activity of the contractile protein actin has been determined from surface tension measurements using the Wilhelmy hanging-plate method. Actin, a very soluble protein, moves from the subphase to the air-water interface to make a film. In the absence of magnesium, actin is monomeric and is known as G-actin. During the compression the monomers change their conformation or orientation at the interface and they are then pushed reversibly into the subphase upon further compression. No collapse occurs. Actin monomers in the presence of magnesium become activated; at concentrations greater than some critical value, actin polymerizes to form filaments of F-actin. The actin filaments have a higher surface activity than the actin monomers either because they are more hydrophobic or because F-actin, a rigid polymer, is much more efficient at creating excluded volume. The actin filaments then form a rigid film at the interface that collapses when the surface area is decreased. At less than the critical concentration, the actin monomers are present in the subphase in their activated form. However, their concentration increases at the interface during film compression until the critical concentration is reached. The surface pressure isotherm in this case has the characteristics of a G-actin film at the beginning of the compression and of an F-actin film at the end of the compression process.     

Self-assembled monolayers of globotriaosylceramide (Gb3) mimics: surface-specific affinity with shiga toxins

Self-assembled monolayers (SAMs) of Gb3 mimics having different lengths of alkyl chains were prepared on gold surfaces, and their interactions with galactose-specific lectin (RCA(120)) and Shiga toxins (Stxs) were investigated by a quartz crystal microbalance (QCM) in aqueous solutions. Their interaction with RCA(120) was enhanced owing to the "cluster effect," regardless of the alkyl chain length of the SAMs. The interaction with Stxs was dependent on the alkyl chain length of Gb3 mimics. Stx-1 and Stx-2 showed a stronger affinity to the Gb3C2 SAM with ethyl disulfide and to the Gb3C10 with decyl disulfide, respectively. Gb3 glycoconjugate polymer with no alkyl spacer inhibited the adsorption of Stx-1 to Gb3C10 SAM but did not inhibit the adsorption of Stx-2 to Gb3C10 SAM. The results suggest that the alkyl chain of the glycolipid takes part in the binding to Stx-2 but not to Stx-1, which is also supported by the computer simulation of Stx-1 with a Gb3 model substance.     

Analyzing multi-environment variety trials using randomization-derived mixed models

Of interest is the analysis of results of a series of experiments repeated at several environments with the same set of plant varieties. Suppose that the experiments, multi-environment variety trials, are all conducted in resolvable incomplete block (IB) designs. Following the randomization approach adopted in Caliński and Kageyama (2000, Lecture Notes in Statistics, 150), two models for analyzing such trial data can be considered. One is derived under a complete additivity assumption, the other takes into account possible different responses of the varieties to variable environmental conditions. The analysis under the first, the standard model, does not provide answers to questions related to the performance of the individual varieties at different environments. These can be considered when using the more general second model. The purpose of this article is to devise interesting parameter estimation and hypothesis testing procedures under that more realistic model. Its application is illustrated by a thorough analysis of a set of data from a winter wheat series of trials.     

The evaluation of treatment when center-specific selection criteria vary with respect to patient risk

Many standards of medical care are based on the demonstrated effects of various treatment strategies or processes. Unlike pharmacological treatments, these strategies or processes are not necessarily subjected to rigorous clinical trials and their benefit is frequently assessed from observational data. For evaluating the influence of such medical processes on patient outcomes, not only is risk adjustment an issue, but also the "center effect" represents an important, often overlooked consideration. Both the quality of care and the tendency to use certain treatments or processes vary from one center to another. The induced similarity in outcomes within center, as well as the potential for confounding by center, needs to be addressed within the context of risk adjustment. In addition, center-specific selection criteria for a treatment strategy can vary with respect to patient risk. Because of these considerations, it is important to adequately separate the within-center effects of the treatment or strategy from the across-center effects, which relate more to center performance. The primary objective of this article is to explore and extend current methods of dealing with center confounding for dichotomous outcomes, primarily for the situation where selection on the basis of patient risk can vary from center to center. A simulation study compares results from several different analytic methods and provides evidence for the importance of considering confounding due to both risk and center when evaluating the effectiveness of a process. An example that examines the effect of early extubation after bypass surgery is also presented.     

Temperature and pressure dependent growth and morphology of DMPC/DSPC domains studied by Brewster angle microscopy

In this work, the temperature and pressure dependent growth of domains in DMPC/DSPC monolayers at various molar ratios was studied by Brewster angle microscopy. Upon compression, roughly discoidal domains with some branching are formed. Further compression leads to an increase in both the number and the average size of the domains, which range between ca. 5 and 20 microm. The isobaric heating of the monolayers results in a gradual decrease of the domain size until their disappearance. The size and morphology of the domains depend not only on equilibrium parameters such as temperature, pressure and composition, but appear to be also strongly dependent on non-equilibrium parameters such as the rate of perturbation. The comparison between our results and those previously published for bilayers allows us to infer that the growth behaviour in monolayers can be qualitatively but not quantitatively extrapolated to bilayers.     

Mesenchymal stem cells inhibit lymphocyte proliferation by mitogens and alloantigens by different mechanisms

Human mesenchymal stem cells (MSCs) have immuno-modulatory properties. They inhibit T-cell proliferation to mitogens and alloantigens in vitro and prolong skin graft survival in vivo. We found that MSCs inhibited the proliferation of peripheral blood lymphocytes (PBLs) to phorbol myristate acetate (PMA), suggesting that MSCs exert an inhibitory effect downstream of the receptor level. We analyzed cytokine profiles of PBLs co-cultured with MSCs. MSCs increased interleukin (IL)-2 and soluble IL-2 receptor in mixed lymphocyte cultures (MLCs), while IL-2 and IL-2R decreased in phytohemagglutinin (PHA)-stimulated PBL cultures. MSCs inhibited IL-2 induced proliferation, without absorbing IL-2. IL-10 levels increased in MLCs co-cultured with 10% MSCs, while the levels were not affected in PHA cultures. In MLCs inhibited by MSCs, antibodies against IL-10 further suppressed proliferation but had no effect in PHA cultures. Addition of indomethacin, an inhibitor of prostaglandin-synthesis, restored part of the inhibition by MSCs in PHA cultures. However, indomethacin did not affect MSC-induced inhibition in MLCs. To conclude, our data indicate that MSC-induced suppression is a complex mechanism affecting IL-2 and IL-10 signaling and may function differently, depending on T-cell stimuli. Prostaglandins are important in the inhibition by MSCs when the T cells were activated by PHA, but not alloantigens.