Effects of Cholesterol on Water Permittivity of Biomimetic Ion Pair Amphiphile Bilayers: Interplay between Membrane Bending and Molecular Packing.

Ion pair amphiphile (IPA), a molecular complex composed of a pair of cationic and anionic amphiphiles, is an inexpensive phospholipid substitute to fabricate vesicles with various pharmaceutical applications. Modulating the physicochemical and permeation properties of IPA vesicles are important for carrier designs. Here, we applied molecular dynamics simulations to examine the cholesterol effects on the structures, mechanics, and water permittivity of hexadecyltrimethylammonium-dodecylsulfate (HTMA-DS) and dodecyltrimethylammonium- hexadecylsulfate (DTMA-HS) IPA bilayers. Structural and mechanical analyses indicate that both IPA systems are in gel phase at 298 K. Adding cholesterol induces alkyl chain ordering around the rigid sterol ring and increases the cavity density within the hydrophilic region of both IPA bilayers. Furthermore, the enhanced alkyl chain ordering and the membrane deformation energy induced by cholesterol increase the permeation free energy penalty. In contrast, cholesterol has minor effects on the water local diffusivities within IPA membranes. Overall, the cholesterol reduces the water permittivity of rigid IPA membranes due to the synergistic effects of increased alkyl chain ordering and enhanced membrane mechanical modulus. The results provide molecular insights into the effects of molecular packing and mechanical deformations on the water permittivity of biomimetic IPA membranes, which is critical for designing IPA vesicular carriers.

Characterising the compressive anisotropic properties of analogue bone using optical strain measurement.

The validity of conclusions drawn from pre-clinical tests on orthopaedic devices depends upon accurate characterisation of the support materials: frequently, polymer foam analogues. These materials often display anisotropic mechanical behaviour, which may considerably influence computational modelling predictions and interpretation of experiments. Therefore, this study sought to characterise the anisotropic mechanical properties of a range of commonly used analogue bone materials, using non-contact multi-point optical extensometry method to account for the effects of machine compliance and uneven loading. Testing was conducted on commercially available 'cellular', 'solid' and 'open-cell' Sawbone blocks with a range of densities. Solid foams behaved largely isotropically. However, across the available density range of cellular foams, the average Young's modulus was 23%-31% lower (p < 0.005) perpendicular to the foaming direction than parallel to it, indicating elongation of cells with foaming. The average Young's modulus of open-celled foams was 25%-59% higher (p < 0.05) perpendicular to the foaming direction than parallel to it. This is thought to result from solid planes of material that were observed perpendicular to the foaming direction, stiffening the bulk material. The presented data represent a reference to help researchers design, model and interpret tests using these materials.

Interplay between alkyl chain asymmetry and cholesterol addition in the rigid ion pair amphiphile bilayer systems.

Ion pair amphiphile (IPA), a molecular complex composed of a pair of cationic and anionic surfactants, has been proposed as a novel phospholipid substitute. Controlling the physical stability of IPA vesicles is important for its application developments such as cosmetic and drug deliveries. To investigate the effects of IPA alkyl chain combinations and the cholesterol additive on the structural and mechanical properties of IPA vesicular bilayers, we conducted a series of molecular dynamics studies on the hexadecyltrimethylammonium-dodecylsulfate (HTMA-DS) and dodecyltrimethylammonium-hexadecylsulfate (DTMA-HS) IPA bilayers with cholesterol. We found that both IPA bilayers are in the gel phase at 298 K, consistent with experimental observations. Compared with the HTMA-DS system, the DTMA-HS bilayer has more disordered alkyl chains in the hydrophobic region. When adding cholesterol, it induces alkyl chain ordering around its rigid sterol ring. Yet, cholesterol increases the molecular areas for all species and disturbs the molecular packing near the hydrophilic region and the bilayer core. Cholesterol also promotes the alkyl chain mismatch between the IPA moieties, especially for the DTMA-HS bilayer. The combined effects lead to non-monotonically enhancement of the membrane mechanical moduli for both IPA-cholesterol systems. Furthermore, cholesterol can form H-bonds with the alkylsulfate and thus enhance the contribution of alkylsulfate to the overall mechanical moduli. Combined results provide valuable molecular insights into the roles of each IPA component and the cholesterol on modulating the IPA bilayer properties.

Pandoraea sp. RB-44, a novel quorum sensing soil bacterium.

Proteobacteria are known to communicate via signaling molecules and this process is known as quorum sensing. The most commonly studied quorum sensing molecules are N-acylhomoserine lactones (AHLs) that consists of a homoserine lactone moiety and an N-acyl side chain with various chain lengths and degrees of saturation at the C-3 position. We have isolated a bacterium, RB-44, from a site which was formally a landfill dumping ground. Using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry analysis, this isolate was identified as a Pandoraea sp.which was then screened for AHL production using biosensors which indicated its quorum sensing properties. To identify the AHL profile of Pandoraea sp. RB-44, we used high resolution tandem mass spectrometry confirming that this isolate produced N-octanoylhomoserine lactone (C8-HSL). To the best of our knowledge, this is the first report that showed quorum sensing activity exhibited by Pandoraea sp. Our data add Pandoraea sp. to the growing number of bacteria that possess QS systems.

Aeromonas aquariorum clinical isolates: antimicrobial profiles, plasmids and genetic determinants.

The objective of this study was to investigate the antimicrobial resistance patterns of 47 clinical isolates of Aeromonas aquariorum and to identify the presence of plasmids and the relevant antibiotic resistance genes (ARGs). Antibiotic susceptibilities were determined by the standard disc diffusion method. The presence of plasmids and ARGs was detected by gel electrophoresis and monoplex PCR. Resistance to amoxicillin/clavulanic acid (98%), amoxicillin (91%), gentamicin (13%), trimethoprim/sulfamethoxazole (11%) and kanamycin (6%) was observed, whilst no ciprofloxacin- or amikacin-resistant strains were detected. All isolates harboured plasmids with sizes ranging from ca. 2 kb to 10 kb. PCR revealed that A. aquariorum carried three ß-lactam resistance genes (bla(TEM), bla(MOX) and bla(PSE)) and two sulphonamide resistance genes (sul1 and sul2). This study provides further understanding of the phenotypic and genotypic characteristics of multiresistant A. aquariorum clinical isolates.

A computational model of amoeboid cell migration.

We present a two-dimensional computational model of amoeboid cell migration characterised by cell shape changes due to the formation and extension of protrusions known as blebs. Using this model, we numerically study the deformation of the cell membrane during blebbing, as well as the effects of obstacles, such as protein fibres in the extracellular matrix, on the motion of the blebbing cell. The model is established in the framework of Stokes flow. Cell membrane deformation is coupled to membrane tension, membrane bending, membrane-cortex adhesion and cortical activities via the intracellular and extracellular fluid field described by the Stokes equation. By assuming that actin monomers move at constant speed towards the membrane and polymerise when they approach the membrane, our model shows that the cell movement in unconfined space can be sustained. We also study how a migrating cell interacts with obstacles hydrodynamically, allowing us to model cell migration in confined environments and to investigate the effects of confinement on the cell migration speed. Our model can be used to further study how tumour cells move through the extracellular matrix during cancer metastasis.

HepG2 cells infected with Klebsiella pneumoniae show DNA laddering at apoptotic and necrotic stages.

Virulent Klebasiella pneumoniae (KP) inflicts severe liver abscesses in infected patients. This study investigated how the bacterial infection affected cell survival at the molecular level, in a cultured cell model. A strain of KP highly virulent in mice was isolated from a patient with liver abscess, and was used to infect HepG2 cells. The infected cells were examined for their viability, DNA fragmentation, and proteins involved in apoptosis or necrosis. We found that the infection decreased the viability of HepG2 cells at 4 hours (h) to 12 h post infection (pi). DNA ladders appeared 6-16 h pi and flow cytometry analysis showed apoptosis at 3-5 h pi, secondary necrosis at 6-9 h pi and primary necrosis at 8-9 h pi. Cleavages of Caspase 7, Caspase 9, α-Fodrin, and PARP were evident at 2-4 h pi. At 7 h pi, we observed the following: increased nuclear AIF, the release from mitochondria of cytosolic Apaf-1 and Cyt c, increased DFF40 expression, decreased DFF45, decreased BcL-xL and the release of Endo G from mitochondria to nucleus. Cellular ATP concentration decreased at 4-8 h pi, accompanied by increased Calpain-2 expression. In summary, infected HepG2 cells underwent apoptosis early after infection and progressed to secondary necrosis and primary necrosis. Nuclear fragmentation corresponded to Caspase 7 activation and the appearance of Endo G and DFF40 in the nucleus, with a concomitant decrease in DFF45. Mitochondrial release of Cyt c together with activation of Caspase 9 and Apaf-1 in cytosol was also observed. Early-hour cleavage of poly(ATP-ribose) polymerase (PARP) followed by the later activation of PARP corresponded to the appearance of DNA laddering, and the depletion of cellular ATP was associated with the appearance of necrosis.

Alum interaction with dendritic cell membrane lipids is essential for its adjuvanticity.

As an approved vaccine adjuvant for use in humans, alum has vast health implications, but, as it is a crystal, questions remain regarding its mechanism. Furthermore, little is known about the target cells, receptors, and signaling pathways engaged by alum. Here we report that, independent of inflammasome and membrane proteins, alum binds dendritic cell (DC) plasma membrane lipids with substantial force. Subsequent lipid sorting activates an abortive phagocytic response that leads to antigen uptake. Such activated DCs, without further association with alum, show high affinity and stable binding with CD4(+) T cells via the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1). We propose that alum triggers DC responses by altering membrane lipid structures. This study therefore suggests an unexpected mechanism for how this crystalline structure interacts with the immune system and how the DC plasma membrane may behave as a general sensor for solid structures.

Antibiograms and molecular subtypes of methicillin-resistant Staphylococcus aureus in local teaching hospital, Malaysia.

The objectives of this study were to determine the antibiotypes, SCCmec subtypes, PVL carriage, and genetic diversity of MRSA strains from a tertiary hospital. Sixtysix MRSA strains were selected randomly (2003, 2004, and 2007) and tested for the Panton-Valentine leukocidin gene, mecA gene, and SCCmec type via a PCR. The antibiograms were determined using a standard disc diffusion method, and the genetic diversity of the isolates was determined by PFGE. Thirty-four antibiograms were obtained, with 55% of the 66 strains exhibiting resistance to more than 4 antimicrobials. All the isolates remained susceptible to vancomycin, and low resistance rates were noted for fusidic acid (11%), rifampicin (11%), and clindamycin acid (19%). The MRSA isolates that were multisensitive (n=12) were SCCmec type IV, whereas the rest (multiresistant) were SCCmec type III. Only two isolates (SCCmec type IV) tested positive for PVL, whereas all the isolates were mecA-positive. The PFGE was very discriminative and subtyped the 66 isolates into 55 pulsotypes (F=0.31-1.0). The multisensitive isolates were distinctly different from the multidrug-resistant MRSA. In conclusion, no vancomycin-resistant isolate was observed. The Malaysian MDR MRSA isolates were mostly SCCmec type III and negative for PVL. These strains were genetically distinct from the SCCmec type IV strains, which were sensitive to SXT, tetracycline, and erythromycin. Only two strains were SCCmec IV and PVL-positive. The infections in the hospital concerned were probably caused by multiple subtypes of MRSA.

Numerical methods for computing the ground state of spin-1 Bose-Einstein condensates in a uniform magnetic field.

We propose efficient and accurate numerical methods for computing the ground-state solution of spin-1 Bose-Einstein condensates subjected to a uniform magnetic field. The key idea in designing the numerical method is based on the normalized gradient flow with the introduction of a third normalization condition, together with two physical constraints on the conservation of total mass and conservation of total magnetization. Different treatments of the Zeeman energy terms are found to yield different numerical accuracies and stabilities. Numerical comparison between different numerical schemes is made, and the best scheme is identified. The numerical scheme is then applied to compute the condensate ground state in a harmonic plus optical lattice potential, and the effect of the periodic potential, in particular to the relative population of each hyperfine component, is investigated through comparison to the condensate ground state in a pure harmonic trap.