Identification of biofilm proteins in non-typeable Haemophilus Influenzae

BACKGROUND: Non-typeable Haemophilus influenzae biofilm formation is implicated in a number of chronic infections including otitis media, sinusitis and bronchitis. Biofilm structure includes cells and secreted extracellular matrix that is "slimy" and believed to contribute to the antibiotic resistant properties of biofilm bacteria. Components of biofilm extracellular matrix are largely unknown. In order to identify such biofilm proteins an ex-vivo biofilm of a non-typeable Haemophilus influenzae isolate, originally from an otitis media patent, was produced by on-filter growth. Extracellular matrix fraction was subjected to proteomic analysis via LC-MS/MS to identify proteins. RESULTS: 265 proteins were identified in the extracellular matrix sample. The identified proteins were analyzed for COG grouping and predicted cellular location via the TMHMM and SignalP predictive algorithms. The most over-represented COG groups identified compared to their frequency in the Haemophilus influenzae genome were cell motility and secretion (group N) followed by ribosomal proteins of group J. A number of hypothetical or un-characterized proteins were observed, as well as proteins previously implicated in biofilm function. CONCLUSION: This study represents an initial approach to identifying and cataloguing numerous proteins associated with biofilm structure. The approach can be applied to biofilms of other bacteria to look for commonalities of expression and obtained information on biofilm protein expression can be used in multidisciplinary approaches to further understand biofilm structure and function

Microwave Reflection and Dielectric Properties of Mortar Subjected to Compression Force and Cyclically Exposed to Water and Sodium Chloride Solution

Corrosion of the reinforcing steel is a major cause of damage and deterioration in reinforced concrete structures such as concrete bridge decks and columns. Chloride intrusion into concrete can lead to depassivation of the steel and initiation of corrosion. Thus, it is very important to be able to nondestructively detect and evaluate the free chloride content in concrete. Near-field microwave nondestructive testing techniques, using open-ended rectangular waveguide probes, have shown great potential for evaluating various properties of concrete, including the successful detection of sodium chloride added to mortar mixing water. In this study, several mortar samples are cyclically soaked in distilled and salt water while also experiencing compression force. Compression force, simulating in-service loading, causes microcracking, which results in increased microcracking and permeability, promoting chloride ingress. The daily microwave reflection and dielectric properties of these samples were measured at 3 GHz. The results show the capability of these microwave measurements for detecting the increased level of chloride permeation and loading as a function of the increasing number of soaking cycles. The influence of salt ingress is shown to be more prominent in the loss factor, while the effect of loading is more evident in the permittivity of the samples

Identification of IbeR as a Stationary-Phase Regulator in Meningitic Escherichia coli K1 that Carries a Loss-of-Function Mutation in rpoS

IbeR is a regulator present in meningitic Escherichia coli strain E44 that carries a loss-of-function mutation in the stationary-phase (SP) regulatory gene rpoS. In order to determine whether IbeR is an SP regulator in E44, two-dimensional gel electrophoresis and LC-MS were used to compare the proteomes of a noninvasive ibeR deletion mutant BR2 and its parent strain E44 in the SP. Four up-regulated (TufB, GapA, OmpA, AhpC) and three down-regulated (LpdA, TnaA, OpmC) proteins in BR2 were identified when compared to E44. All these proteins contribute to energy metabolism or stress resistance, which is related to SP regulation. One of the down-regulated proteins, tryptophanase (TnaA), which is regulated by RpoS in other E. coli strains, is associated with SP regulation via production of a signal molecule indole. Our studies demonstrated that TnaA was required for E44 invasion, and that indole was able to restore the noninvasive phenotype of the tnaA mutant. The production of indole was significantly reduced in BR2, indicating that ibeR is required for the indole production via tnaA. Survival studies under different stress conditions indicated that IbeR contributed to bacteria stress resistance in the SP. Taken together, IbeR is a novel regulator contributing to the SP regulation

EPA/NMED/LANL 1998 water quality results: Statistical analysis and comparison to regulatory standards

Four governmental agencies conducted a round of groundwater, surface water, and spring water sampling at the Los Alamos National Laboratory during 1998. Samples were split among the four parties and sent to independent analytical laboratories. Results from three of the agencies were available for this study. Comparisons of analytical results that were paired by location and date were made between the various analytical laboratories. The results for over 50 split samples analyzed for inorganic chemicals, metals, and radionuclides were compared. Statistical analyses included non-parametric (sign test and signed-ranks test) and parametric (paired t-test and linear regression) methods. The data pairs were tested for statistically significant differences, defined by an observed significance level, or p-value, less than 0.05. The main conclusion is that the laboratories' performances are similar across most of the analytes that were measured. In some 95% of the laboratory measurements there was agreement on whether contaminant levels exceeded regulatory limits. The most significant differences in performance were noted for the radioactive suite, particularly for gross alpha particle activity and Sr-90

Microwave spectrum and structure of ethylene ozonide: Effects of large axes rotations in structure calculations

Rotational spectra for 14 isotopic species of ethylene ozonide have now been assigned. The consistency of the Kraitchman substitution structure was checked by calculating the Op---Op bond distance six ways; the values ranged from 1.458 to 1.502 A. This variation was attributed to an amplification of residual vibrational effects by large axes rotations upon isotopic substitution. Estimates of errors produced from this effect were made and a procedure was developed for choosing rs parameters in which the effect is minimized. This gave the following ring parameters: d(COe) = 1.416 A, d(COp) = 1.412 A, d(OO) = 1.461 A, eC = 104.8[deg], eCOp = 105.5[deg], pOp = 99.3[deg].Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/21905/1/0000312.pd

A direct path to dependable software

What would it take to make software more dependable? Until now, most approaches have been indirect: some practices – processes, tools or techniques – are used that are believed to yield dependable software, and the argument for dependability rests on the extent to which the developers have adhered to them. This article argues instead that developers should produce direct evidence that the software satisfies its dependability claims. The potential advantages of this approach are greater credibility (since the argument is not contingent on the effectiveness of the practices) and reduced cost (since development resources can be focused where they have the most impact)

Phase transitions in biological membranes

Native membranes of biological cells display melting transitions of their lipids at a temperature of 10-20 degrees below body temperature. Such transitions can be observed in various bacterial cells, in nerves, in cancer cells, but also in lung surfactant. It seems as if the presence of transitions slightly below physiological temperature is a generic property of most cells. They are important because they influence many physical properties of the membranes. At the transition temperature, membranes display a larger permeability that is accompanied by ion-channel-like phenomena even in the complete absence of proteins. Membranes are softer, which implies that phenomena such as endocytosis and exocytosis are facilitated. Mechanical signal propagation phenomena related to nerve pulses are strongly enhanced. The position of transitions can be affected by changes in temperature, pressure, pH and salt concentration or by the presence of anesthetics. Thus, even at physiological temperature, these transitions are of relevance. There position and thereby the physical properties of the membrane can be controlled by changes in the intensive thermodynamic variables. Here, we review some of the experimental findings and the thermodynamics that describes the control of the membrane function.Comment: 23 pages, 15 figure