Thermodynamics of polymer adsorption to a flexible membrane

We analyze the structural behavior of a single polymer chain grafted to an attractive, flexible surface. Our model is composed of a coarse-grained bead-and-spring polymer and a tethered membrane. By means of extensive parallel tempering Monte Carlo simulations it is shown that the system exhibits a rich phase behavior ranging from highly ordered, compact to extended random coil structures and from desorbed to completely adsorbed or even partially embedded conformations. These findings are summarized in a pseudophase diagram indicating the predominant class of conformations as a function of the external parameters temperature and polymer-membrane interaction strength. By comparison with adsorption to a stiff membrane surface it is shown that the flexibility of the membrane gives rise to qualitatively new behavior such as stretching of adsorbed conformations

Partial Characterization of the Major Lipooligosaccharide from a Strain of Haemophilus ducreyi, the Causative Agent of Chancroid, a Genital Ulcer Disease

The first preliminary structure of a surface lipooligosaccharide from Haemophilus ducreyi has been determined. The major oligosaccharide was released by mild acid hydrolysis and analyzed by liquid secondary ion and tandem mass spectrometry. The mass spectral data combined with composition and methylation analysis yielded the most probable structure; Gal1----4GlcNAc1----3Gal1----4Hep1----6Glc1----( Hep1----2Hep1----)3,4Hep1---- KDO, where the reducing terminal 3-deoxy-D-manno-octulosonic acid (or KDO) exists in an anhydro form. This anhydro species results from the elimination of a phosphate from C-4 of KDO during mild acid hydrolysis. The core heptose trisaccharide consists of L-glycero-D-manno-heptose, but analysis of the peracetylated sugars indicated that the 1,4-linked heptose is likely D-glycero-D-manno-heptose. The monoclonal antibody 3F11 generated against Neisseria gonorrhoeae also binds to this lipooligosaccharide and suggests that the terminal trisaccharide is Gal beta 1----4GlcNAc beta 1----3Gal beta 1----, an epitope found in the glycose moiety of the human erythrocyte glycosphingolipid lactoneotetraglycosylceramide. Mass spectrometric and composition analysis of the lipid A moiety shows that it is similar to the lipid A of Haemophilus influenzae strain I-69 Rd-/b+ proposed by Helander et al. (Helander, I. M., Lindner, B., Brade, H., Altmann, K., Lindberg, A. A., Rietschel, E. T., and Zähringer, U. (1988) Eur. J. Biochem. 177, 483-492). Electrospray mass spectrometric analysis of the intact O-deacylated lipooligosaccharides gave an average Mr of 2710, and supported an overall structure consisting of the above nonasaccharide linked directly to a diphosphorylated lipid A moiety through the single KDO which is phosphorylated. This structure should provide a framework to investigate the roles of lipooligosaccharides in the host immunochemical response and pathology of H. ducreyi infection, a leading cause of genital ulcer disease

Population-Sequencing as a Biomarker of Burkholderia mallei and Burkholderia pseudomallei Evolution through Microbial Forensic Analysis

Large-scale genomics projects are identifying biomarkers to detect human disease. B. pseudomallei and B. mallei are two closely related select agents that cause melioidosis and glanders. Accurate characterization of metagenomic samples is dependent on accurate measurements of genetic variation between isolates with resolution down to strain level. Often single biomarker sensitivity is augmented by use of multiple or panels of biomarkers. In parallel with single biomarker validation, advances in DNA sequencing enable analysis of entire genomes in a single run: population-sequencing. Potentially, direct sequencing could be used to analyze an entire genome to serve as the biomarker for genome identification. However, genome variation and population diversity complicate use of direct sequencing, as well as differences caused by sample preparation protocols including sequencing artifacts and mistakes. As part of a Department of Homeland Security program in bacterial forensics, we examined how to implement whole genome sequencing (WGS) analysis as a judicially defensible forensic method for attributing microbial sample relatedness; and also to determine the strengths and limitations of whole genome sequence analysis in a forensics context. Herein, we demonstrate use of sequencing to provide genetic characterization of populations: direct sequencing of populations

The Moraxella adhesin UspA1 binds to its human CEACAM1 receptor by a deformable trimeric coiled-coil

Moraxella catarrhalis is a ubiquitous human-specific bacterium commonly associated with upper and lower respiratory tract infections, including otitis media, sinusitis and chronic obstructive pulmonary disease. The bacterium uses an autotransporter protein UspA1 to target an important human cellular receptor carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1). Using X-ray crystallography, we show that the CEACAM1 receptor-binding region of UspA1 unusually consists of an extended, rod-like left-handed trimeric coiled-coil. Mutagenesis and binding studies of UspA1 and the N-domain of CEACAM1 have been used to delineate the interacting surfaces between ligand and receptor and guide assembly of the complex. However, solution scattering, molecular modelling and electron microscopy analyses all indicate that significant bending of the UspA1 coiled-coil stalk also occurs. This explains how UspA1 can engage CEACAM1 at a site far distant from its head group, permitting closer proximity of the respective cell surfaces during infection

Entry of Yersinia pestis into the Viable but Nonculturable State in a Low-Temperature Tap Water Microcosm

Yersinia pestis, the causative agent of plague, has caused several pandemics throughout history and remains endemic in the rodent populations of the western United States. More recently, Y. pestis is one of several bacterial pathogens considered to be a potential agent of bioterrorism. Thus, elucidating potential mechanisms of survival and persistence in the environment would be important in the event of an intentional release of the organism. One such mechanism is entry into the viable but non-culturable (VBNC) state, as has been demonstrated for several other bacterial pathogens. In this study, we showed that Y. pestis became nonculturable by normal laboratory methods after 21 days in a low-temperature tap water microcosm. We further show evidence that, after the loss of culturability, the cells remained viable by using a variety of criteria, including cellular membrane integrity, uptake and incorporation of radiolabeled amino acids, and protection of genomic DNA from DNase I digestion. Additionally, we identified morphological and ultrastructural characteristics of Y. pestis VBNC cells, such as cell rounding and large periplasmic spaces, by electron microscopy, which are consistent with entry into the VBNC state in other bacteria. Finally, we demonstrated resuscitation of a small number of the non-culturable cells. This study provides compelling evidence that Y. pestis persists in a low-temperature tap water microcosm in a viable state yet is unable to be cultured under normal laboratory conditions, which may prove useful in risk assessment and remediation efforts, particularly in the event of an intentional release of this organism

Tribological properties of biodegradable polyurethanes of various structure and content of rigid elements

Poliuretany (PUR) o zróżnicowanej budowie chemicznej i różnej zawartości segmentów sztywnych zsyntetyzowano przy użyciu 4,4'-diizocyjanianu dicykloheksylenometanu (HMDI), polikaprolaktanodiolu (PCL) oraz glikolu etylenowego (GE) i glicerolu (G) jako przedłużaczy łańcucha. Otrzymano pięć różnych typów PUR. Ich właściwości fizykochemiczne oceniano metodami: spektroskopii w podczerwieni, różnicowej kalorymetrii skaningowej (DSC) i termograwimetrii (TG). Badano twardość Shore'a i wytrzymałość przy rozciąganiu wytworzonych PUR oraz określano ich charakterystyki tribologiczne. Wyniki badań potwierdziły wpływ budowy i zawartości segmentów sztywnych w makrocząsteczkach PUR na ich właściwości, zwłaszcza na charakterystyki tribologiczne.Five polyurethanes (PUR) of different chemical structure and variable content of rigid elements have been synthesized using 4,4'-dicyclohexylmethane diisocyanate (HMDI), polycaprolactone diol (PCL) as well as ethylene glycol (GE) and glycerol (G) as chain extenders. The physicochemical properties of the obtained PUR have been investigated with the aid of infrared spectroscopy, differential scanning calorimetry (DSC) and thermogravimetry (TG). The mechanical properties (Shore hardness and tensile strength) and tribological characteristics (pin-on-disc tribometer) of PUR have been determined. The results of the study confirm the influence of structure and rigid segment content on the properties of PUR macromolecules, particularly their tribological characteristics

Inactivation of the Moraxella catarrhalis Superoxide Dismutase SodA Induces Constitutive Expression of Iron-Repressible Outer Membrane Proteins

Many pathogens produce one or more superoxide dismutases (SODs), enzymes involved in the detoxification of endogenous and exogenous reactive oxygen species that are encountered during the infection process. One detectable cytoplasmic SOD was identified in the human mucosal pathogen Moraxella catarrhalis, and the gene responsible for the SOD activity, sodA, was isolated from a recent pediatric clinical isolate (strain 7169). Sequence analysis of the cloned M. catarrhalis 7169 DNA fragment revealed an open reading frame of 618 bp encoding a polypeptide of 205 amino acids with 48 to 67% identity to known bacterial manganese-cofactored SODs. An isogenic M. catarrhalis sodA mutant was constructed in strain 7169 by allelic exchange. In contrast to the wild-type 7169, the 7169::sodK20 mutant was severely attenuated for aerobic growth, even in rich medium containing supplemental amino acids, and exhibited extreme sensitivity to the redox-active agent methyl viologen. The ability of recombinant SodA to rescue the aerobic growth defects of E. coli QC774, a sodA sodB-deficient mutant, demonstrated the functional expression of SOD activity by cloned M. catarrhalis sodA. Indirect SOD detection assays were used to visualize both native and recombinant SodA activity in bacterial lysates. This study demonstrates that M. catarrhalis SodA plays a critical role in the detoxification of endogenous, metabolically produced oxygen radicals. In addition, the outer membrane protein (OMP) profile of 7169::sodK20 was consistent with iron starvation in spite of growth under iron-replete conditions. This novel observation indicates that M. catarrhalis strains lacking SodA constitutively express immunogenic OMPs previously described as iron repressible, and this potentially attenuated mutant strain may be an attractive vaccine candidate

Tribological properties of resorbable polylactide-based biomaterials

Celem pracy było przeprowadzenie badań tribologicznych i ocena zużycia biokompozytów polimerowych. Do badań przygotowano trzy rodzaje kompozytów z polilaktydu modyfikowanego dodatkami w postaci: włókien węglowych, fosforanu trójwapnia oraz hydroksyapatytu. Ich zachowanie w warunkach tarciowych porównywano z próbkami referencyjnymi (PLA). Badania tribologiczne realizowano w warunkach tarcia suchego oraz w obecności smaru – soli fizjologicznej (roztwór wodny 0,9% NaCl). Na podstawie badań stwierdzono, że wprowadzenie modyfikatorów ceramicznych do osnowy polilaktydowej poprawiło charakterystyki tribologiczne, zaś dodatek włókien węglowych wpłynął najkorzystniej na obniżenie współczynnika tarcia (μ). Stwierdzono istotny wpływ środka smarnego. O ile w obecności smaru wartości μ dla wszystkich badanych materiałów obniżyły się, to zużycie - wyznaczone na podstawie zmian masy - zmniejszyło się tylko dla czystego polilaktydu oraz polilaktydu modyfikowanego włóknem węglowym. W przypadku kompozytów z fosforanem trójwapniowym i hydroksyapatytem zarejestrowano intensyfikację zużycia. Uzupełnieniem przeprowadzonych badań były obserwacje powierzchni testowanych próbek przy pomocy elektronowego mikroskopu skaningowego (SEM). Niezależnie od warunków (ze smarem czy też bez) najmniejszą odpornością na zużycie tarciowe cechował się czysty polilaktyd. Kompozyty z dodatkami ceramicznymi wykazywały wyraźne ślady zużycia, zaś obserwacje powierzchni kompozytów wzmacnianych włóknami węglowymi ujawniły, że zewnętrzne warstwy poddawane oddziaływaniu tarcia narażone są na fragmentacje włókien węglowych. Zjawisko to może mieć niekorzystny wpływ na możliwość aplikacji takich materiałów w warunkach pracy, w których są narażone na ścieranie.The objective of this study was the observation and evaluation of tribological characteristics and behavior of biocomposites with polymer matrix under different friction conditions. Three types of polylactide composites were considered: with addition of carbon fibers (CF), tricalcium phosphate (TCP) and hydroxyapatite (HAP). As a reference pure polylactide (PLA) samples were prepared. Tribological tests were conducted under dry friction conditions and in the presence of a lubricant – physiological saline (0.9% NaCl aqueous solution). The results clearly show that introduction of ceramic modifiers to the polylactide matrix enhanced tribological resistance of the composite and carbon fibers addition reduced the value of friction coefficient (μ). It was observed that when the lubricant was present lower values of μ were recorded for all composites but when the wear process was analyzed it was revealed that the loss of mass was reduced only for pure PLA and PLA modified with carbon fibers. For ceramic modified composites the wear processes were more intense. Scanning electron microscopy was used for surface observations of tested samples. Regardless of friction conditions (with or without lubricant) pure polylactide samples, as the softest, wear the most. When surface of composites with ceramic additives were analyzed the biggest grooves and hollows ware observed. Samples with CF exposed to friction conditions reveal inclination to fibers fragmentation, thus application of such composites in kinematic joints in biomedical devices is limited