Estimation of anisotropic permeability in trabecular bone based on microCT imaging and pore-scale fluid dynamics simulations.

In this paper, a comprehensive framework is proposed to estimate the anisotropic permeability matrix in trabecular bone specimens based on micro-computed tomography (microCT) imaging combined with pore-scale fluid dynamics simulations. Two essential steps in the proposed methodology are the selection of (i) a representative volume element (RVE) for calculation of trabecular bone permeability and (ii) a converged mesh for accurate calculation of pore fluid flow properties. Accurate estimates of trabecular bone porosities are obtained using a microCT image resolution of approximately 10 µm. We show that a trabecular bone RVE in the order of 2 × 2 × 2 mm3 is most suitable. Mesh convergence studies show that accurate fluid flow properties are obtained for a mesh size above 125,000 elements. Volume averaging of the pore-scale fluid flow properties allows calculation of the apparent permeability matrix of trabecular bone specimens. For the four specimens chosen, our numerical results show that the so obtained permeability coefficients are in excellent agreement with previously reported experimental data for both human and bovine trabecular bone samples. We also identified that bone samples taken from long bones generally exhibit a larger permeability in the longitudinal direction. The fact that all coefficients of the permeability matrix were different from zero indicates that bone samples are generally not harvested in the principal flow directions. The full permeability matrix was diagonalized by calculating the eigenvalues, while the eigenvectors showed how strongly the bone sample's orientations deviated from the principal flow directions. Porosity values of the four bone specimens range from 0.83 to 0.86, with a low standard deviation of ± 0.016, principal permeability values range from 0.22 to 1.45 ⋅ 10 -8 m2, with a high standard deviation of ± 0.33. Also, the anisotropic ratio ranged from 0.27 to 0.83, with high standard deviation. These results indicate that while the four specimens are quite similar in terms of average porosity, large variability exists with respect to permeability and specimen anisotropy. The utilized computational approach compares well with semi-analytical models based on homogenization theory. This methodology can be applied in bone tissue engineering applications for generating accurate pore morphologies of bone replacement materials and to consistently select similar bone specimens in bone bioreactor studies.

The minimal leak test technique for endotracheal cuff maintenance.

Endotracheal tube (ETT) cuff pressure management is an essential part of airway management in intubated and mechanically ventilated patients. Both under- and over-inflation of the ETT cuff can lead to patient complications, with an ideal pressure range of 20-30 cmH2O defined. A range of techniques are employed to ensure adequate ETT cuff inflation, with little comparative data. We performed an observational cross-sectional study in a tertiary metropolitan ICU, assessing the relationship between the minimal leak test and cuff manometry. Forty-five mechanically ventilated patients, over a three-month period, had ETT cuff manometry performed at the same time as their routine cuff maintenance (minimal leak test). Bedside nurse measurements were compared with investigator measurements. At the endpoint of cuff inflation, 20 of 45 patients (44%) had cuff pressures between 20 and 30 cmH2O; 11 of 45 patients (24%) had cuff pressures <20 cmH2O; 14 of 45 patients (31%) had cuff pressures ≥30 cmH2O. Univariate analysis demonstrated an association between both patient obesity and female gender requiring less ETT cuff volume (P=0.008 and P <0.001 respectively), though this association was lost on multivariate analysis. No association was demonstrated between any measured variables and cuff pressures. Inter-operator reliability in performing the minimal leak test showed no evidence of bias between nurse and investigators (Pearson coefficient = 0.897). We conclude the minimal leak test for maintenance of ETT cuffs leads to both over- and under-inflation, and alternative techniques, such as cuff manometry, should be employed.

Electrolytic drops in an electric field: A numerical study of drop deformation and breakup.

The deformation and breakup of an axisymmetric, conducting drop suspended in a nonconducting medium and subjected to an external electric field is numerically investigated here using an electrokinetic model. This model uses a combined level set-volume of fluid formulation of the deformable surfaces, along with a multiphase implementation of the Nernst-Planck equation for transport of ions, that allows for varying conductivity inside the drop. A phase diagram, based on a parametric study, is used to characterize the stability conditions. Stable drops with lower ion concentration are characterized by longer drop shapes than those achieved at higher ion concentrations. For higher drop ion concentration, greater charge accumulation is observed at drop tips. Consequently, such drops break up by pinching off rather than tip streaming. The charge contained in droplets released from unstable drops is shown to increase with drop ion concentration. These dynamic drop behaviors depend on the strength of the electric field and the concentration of ions in the drop and result from the interplay between the electric forces arising from the permittivity jump at the drop interface and the ions in the bulk.

Electroviscous resistance of nanofluidic bends.

Analysis tools that quantify the pressure and potential changes occurring over pressure-driven electrokinetic device elements are necessary for the design of optimal laboratory-on-a-chip devices. In this study, the resistance of a nanofluidic silica channel with negatively charged walls containing a 90^{∘} bend to the electroviscous flow of a potassium chloride salt solution is quantified in terms of two equivalent lengths using numerical analysis. One equivalent length is based on the excess pressure drop and the other on the excess potential rise. Over the entire range of simulations conducted, these equivalent lengths are relatively independent of salt concentration, flow velocity, channel size, and surface charge, remaining within the approximate ranges of 1.3-1.5 for the pressure equivalent length and 0.8-1.05 for the potential equivalent length.

Effect of wall permittivity on electroviscous flow through a contraction.

The electroviscous flow at low Reynolds number through a two-dimensional slit contraction with electric double-layer overlap is investigated numerically for cases where the permittivity of the wall material is significant in comparison with the permittivity of the liquid. The liquid-solid interface is assumed to have uniform surface-charge density. It is demonstrated that a finite wall permittivity has a marked effect on the distribution of ions in and around the contraction, with a significant build-up of counter-ions observed at the back-step. The development length of the flow increases substantially as the wall permittivity becomes significant, meaning that the electric double-layers require a longer distance to develop within the contraction. Consequently, there is a corresponding decrease in the hydrodynamic and electro-potential resistance caused by the contraction. The effect of wall-region width on the flow characteristics is also quantified, demonstrating that the development length increases with increasing wall-region width for widths up to 5 channel widths.

Identification and characterization of an ATP binding cassette L-carnitine transporter in Listeria monocytogenes.

We identified an operon in Listeria monocytogenes EGD with high levels of sequence similarity to the operons encoding the OpuC and OpuB compatible solute transporters from Bacillus subtilis, which are members of the ATP binding cassette (ABC) substrate binding protein-dependent transporter superfamily. The operon, designated opuC, consists of four genes which are predicted to encode an ATP binding protein (OpuCA), an extracellular substrate binding protein (OpuCC), and two membrane-associated proteins presumed to form the permease (OpuCB and OpuCD). The operon is preceded by a potential SigB-dependent promoter. An opuC-defective mutant was generated by the insertional inactivation of the opuCA gene. The mutant was impaired for growth at high osmolarity in brain heart infusion broth and failed to grow in a defined medium. Supplementation of the defined medium with peptone restored the growth of the mutant in this medium. The mutant was found to accumulate the compatible solutes glycine betaine and choline to same extent as the parent strain but was defective in the uptake of L-carnitine. We conclude that the opuC operon in L. monocytogenes encodes an ABC compatible solute transporter which is capable of transporting L-carnitine and which plays an important role in osmoregulation in this pathogen.

The radium century.

Last year, the UK Government agreed that a potentially dangerous legacy of radiation-contaminated land should be dealt with, after decades of ignorance and inaction on the part of the authorities. The cost of this could run to many hundreds of millions of pounds, assuming that the hundreds of sites potentially involved, which date from the birth of the nuclear industry, can actually be identified. This very practical problem cannot be tackled without an understanding of the lost history of the production and use of radium--a history that is now precisely a century in duration.

Two peptides derived from the nerve growth factor precursor are biologically active.

This report provides evidence that the proregion of the NGF precursor protein contains two novel bioactive peptides. The presence of pairs of basic amino acid (aa) residues in the NGF proregion suggests that two or three peptides other than NGF may be generated by proteolytic cleavage. Synthetic peptides of 29 aa (LIP1) and 38aa (LIP2) corresponding to the sequences -71 to -43 and -40 to -3 of the proNGF, respectively, were used in this study. ELISA specific for these two peptides revealed their presence in the rat intestine. LIP1 was localized by immunohistochemistry in endocrine cells of the intestinal epithelium, and LIP2 was immunoprecipitated from an intestinal extract. We also provide evidence for the presence of specific receptors for LIP2 in several cell lines. Scatchard analysis indicated the presence of a low affinity binding site with a Kd of approximately 10(-7) M and a high affinity binding site of 10(-9) M. Cross-linking studies revealed receptor forms of about 140 kD and 93 kD in a prostatic adenocarcinoma cell line. LIP1 and LIP2 induced rapid F-actin redistribution in PC12 cells within 2 min of incubation, which suggests a role of LIP1 and LIP2 in the process of neurite outgrowth. Furthermore, both propeptides induced rapid tyrosine phosphorylation of the Trk protein in both prostatic adenocarcinoma cells and PC12 cells, thus implicating trk in their mechanism of action. These results support our hypothesis that two peptides within the NGF precursor protein are biologically active.

Regulation of NGF, BDNF and LNGFR gene expression in ROS 17/2.8 cells.

The secosteroid hormone 1.25-dihydroxyvitamin D3 (1,25(OH)2D3) has been recently shown to enhance the synthesis of NGF to mouse L929 fibroblasts. In view of the critical role of 1,25(OH)2D3 on bone metabolism, it has been investigated if ROS 17/2.8 osteoblastic cells were able to express the nerve growth factor (NGF) gene and if this process was responsive to 1,25(OH)2D3. Results indicate that these cells respond in a dose-dependent manner to the presence of 1,25(OH)2D3 by an increase in NGF mRNA levels. However, the phorbol ester PMA, previously reported to augment the synthesis of NGF via the recruitment of AP-1 complexes, depressed the expression of the NGF gene in ROS cells. In contrast, the mRNA levels of an NGF-related trophic factor, brain-derived neurotrophic factor (BDNF), was increased by PMA but not following 1,25(OH)2D3 treatment. Binding of 125I-NGF to ROS cells displayed the properties of a low affinity NGF receptor (dissociation constant Kd approximately 10(-9) M). In agreement with this result, the mRNA encoding the low affinity NGF receptor (LNGFR) was detected in ROS 17/2.8 cells, unlike trkA transcripts which encode the high affinity receptor. These data suggest that neurotrophins and their low affinity receptor could play an unsuspected role in bone tissue.

Fatty acid compositions of the major phosphoglycerides from fish neural tissues; (n-3) and (n-6) polyunsaturated fatty acids in rainbow trout (Salmo gairdneri) and cod (Gadus morhua) brains and retinas.

The fatty acid compositions of brain phosphoglycerides from a freshwater fish, the rainbow trout (Salmo gairdneri), and a marine fish, the cod (Gadus morhua), were determined and compared with those from a terrestrial mammal, the rat. Fish brain lipids were characterized by a higher degree of unsaturation encompassing increased percentages of (n-3)PUFA (22∶6 and 20∶5) and lower percentages of (n-6)PUFA (20∶4 and 22∶4). However the distribution of fatty acids and specific PUFA between different phosphoglycerides was essentially similar in rat and fish brain tissue. PE and PS contained the highest percentages of 22∶6(n-3), PI was characterized by higher 18∶0 and 20∶4(n-6)/20∶5(n-3), and PC had higher 16∶0 and the lowest percentage of PUFA in all species. A generally similar pattern was found in the fish retinal phosphoglycerides except that PC was also rich in 22∶6(n-3). Overall trout brain phosphoglycerides were slightly more unsaturated than the cod lipids but with lower (n-3)/(n-6) ratios whereas cod retinal lipids were more unsaturated than the trout retinal lipids.