A quantitative assessment of the content of hematopoietic stem cells in mouse and human endosteal-bone marrow: a simple and rapid method for the isolation of mouse central bone marrow.

BACKGROUND: Isolation of bone marrow cells, including hematopoietic stem cells, is a commonly used technique in both the research and clinical settings. A quantitative and qualitative assessment of cell populations isolated from mouse and human bone marrow was undertaken with a focus on the distribution of hematopoietic cells between the central bone marrow (cBM) and endosteal bone marrow (eBM). METHODS: Two approaches to cBM isolation from the hind legs were compared using the C57BL/6J and BALB/cJ strains of laboratory mice. The content of hematopoietic stem cells in eBM was compared to cBM from mice and human fetal bone marrow using flow cytometry. Enzymatic digestion was used to isolate eBM and its effects on antigen expression was evaluated using flow cytometry. Humanized immunodeficient mice were used to evaluate the engraftment of human precursors in the cBM and eBM and the effects of in vivo maturation on the fetal stem cell phenotype were determined. RESULTS: The two methods of mouse cBM isolation yielded similar numbers of cells from the femur, but the faster single-cut method recovered more cells from the tibia. Isolation of eBM increased the yield of mouse and human stem cells. Enzymatic digestion used to isolate eBM did, however, have a detrimental effect on detecting the expression of the human HSC-antigens CD4, CD90 and CD93, whereas CD34, CD38, CD133 and HLA-DR were unaffected. Human fetal HSCs were capable of engrafting the eBM of immunodeficient mice and their pattern of CD13, CD33 and HLA-DR expression partially changed to an adult pattern of expression about 1 year after transplantation. CONCLUSIONS: A simple, rapid and efficient method for the isolation of cBM from the femora and tibiae of mice is detailed. Harvest of tibial cBM yielded about half as many cells as from the femora, representing 6.4 % and 13 %, respectively, of the total cBM of a mouse based on our analysis and a review of the literature. HSC populations were enriched within the eBM and the yield of HSCs from the mouse and human long bones was increased notably by harvest of eBM.

Macroscale adhesion of gecko setae reflects nanoscale differences in subsurface composition.

Surface energies are commonly used to determine the adhesion forces between materials. However, the component of surface energy derived from long-range forces, such as van der Waals forces, depends on the material's structure below the outermost atomic layers. Previous theoretical results and indirect experimental evidence suggest that the van der Waals energies of subsurface layers will influence interfacial adhesion forces. We discovered that nanometre-scale differences in the oxide layer thickness of silicon wafers result in significant macroscale differences in the adhesion of isolated gecko setal arrays. Si/SiO(2) bilayer materials exhibited stronger adhesion when the SiO(2) layer is thin (approx. 2 nm). To further explore how layered materials influence adhesion, we functionalized similar substrates with an octadecyltrichlorosilane monolayer and again identified a significant influence of the SiO(2) layer thickness on adhesion. Our theoretical calculations describe how variation in the SiO(2) layer thickness produces differences in the van der Waals interaction potential, and these differences are reflected in the adhesion mechanics. Setal arrays used as tribological probes provide the first empirical evidence that the 'subsurface energy' of inhomogeneous materials influences the macroscopic surface forces.

Utility of portable spirometry in a pediatric emergency department in children with acute exacerbation of asthma.

OBJECTIVES: The primary purpose of this study was to determine if portable spirometers can be successfully used in an emergency department (ED) in children with an acute exacerbation of asthma. The secondary purpose of this study was to determine if a validated clinical asthma score (CAS) correlates with the spirometry results in children with an acute exacerbation of asthma. METHODS: Children between the ages of 6 and 17 years who presented to an urban free-standing children's hospital ED with an acute exacerbation of asthma were enrolled in our study. On arrival, the CAS was recorded and then portable spirometry was performed. Attempts were continued until acceptable and reproducible flow loop measurements were obtained or until the patient was unable to perform further attempts. Outcomes included success at spirometry and correlation of spirometry with the CAS. RESULTS: A total of 101 patients were enrolled in this study. Of those patients, only 35 (35%) were able to successfully perform portable spirometry. Successful spirometry attempts were associated with older age (10.4 vs. 8.9, p = .01), lower respiratory rates (24.8 vs. 30.2, p = .001), lower heart rates (110 vs. 124, p = .004), and lower CASs (8.4 vs. 9.7, p = .001). Increasing asthma severity correlated with a decreased likelihood of successfully obtaining a useful forced expiratory volume in 1 second (FEV(1)) measurement (p = .013). Compared with cases of mild asthma, a patient with moderate asthma is 33% less likely to be able to perform spirometry, and a patient with severe asthma 93% less likely to perform spirometry. The CAS correlated poorly with the more objective measure of FEV(1)% predicted in those with mild asthma. CONCLUSION: Many children are incapable of using portable spirometry for the evaluation of acute exacerbations of asthma in the ED. The clinical asthma scoring system demonstrated poor correlation with portable spirometry measurements in terms of severity classification.

Effects of humidity on the mechanical properties of gecko setae.

We tested the hypothesis that an increase in relative humidity (RH) causes changes in the mechanical properties of the keratin of adhesive gecko foot hairs (setae). We measured the effect of RH on the tensile deformation properties, fracture, and dynamic mechanical response of single isolated tokay gecko setae and strips of the smooth lamellar epidermal layer. The mechanical properties of gecko setae were strongly affected by RH. The complex elastic modulus (measured at 5 Hz) of a single seta at 80% RH was 1.2 GPa, only 39% of the value when dry. An increase in RH reduced the stiffness and increased the strain to failure. The loss tangent increased significantly with humidity, suggesting that water absorption produces a transition to a more viscous type of deformation. The influence of RH on the properties of the smooth epidermal layer was comparable with that of isolated seta, with the exception of stress at rupture. These values were two to four times greater for the setae than for the smooth layer. The changes in mechanical properties of setal keratin were consistent with previously reported increases in contact forces, supporting the hypothesis that an increase in RH softens setal keratin, which increases adhesion and friction.

Changes in materials properties explain the effects of humidity on gecko adhesion.

Geckos owe their remarkable stickiness to millions of dry setae on their toes, and the mechanism of adhesion in gecko setae has been the topic of scientific scrutiny for over two centuries. Previously, we demonstrated that van der Waals forces are sufficient for strong adhesion and friction in gecko setae, and that water-based capillary adhesion is not required. However, recent studies demonstrated that adhesion increases with relative humidity (RH) and proposed that surface hydration and capillary water bridge formation is important or even necessary. In this study, we confirmed a significant effect of RH on gecko adhesion, but rejected the capillary adhesion hypothesis. While contact forces of isolated tokay gecko setal arrays increased with humidity, the increase was similar on hydrophobic and hydrophilic surfaces, inconsistent with a capillary mechanism. Contact forces increased with RH even at high shear rates, where capillary bridge formation is too slow to affect adhesion. How then can a humidity-related increase in adhesion and friction be explained? The effect of RH on the mechanical properties of setal ß-keratin has escaped consideration until now. We discovered that an increase in RH softens setae and increases viscoelastic damping, which increases adhesion. Changes in setal materials properties, not capillary forces, fully explain humidity-enhanced adhesion, and van der Waals forces remain the only empirically supported mechanism of adhesion in geckos.

Rate-dependent frictional adhesion in natural and synthetic gecko setae.

Geckos owe their remarkable stickiness to millions of dry, hard setae on their toes. In this study, we discovered that gecko setae stick more strongly the faster they slide, and do not wear out after 30,000 cycles. This is surprising because friction between dry, hard, macroscopic materials typically decreases at the onset of sliding, and as velocity increases, friction continues to decrease because of a reduction in the number of interfacial contacts, due in part to wear. Gecko setae did not exhibit the decrease in adhesion or friction characteristic of a transition from static to kinetic contact mechanics. Instead, friction and adhesion forces increased at the onset of sliding and continued to increase with shear speed from 500 nm s(-1) to 158 mm s(-1). To explain how apparently fluid-like, wear-free dynamic friction and adhesion occur macroscopically in a dry, hard solid, we proposed a model based on a population of nanoscopic stick-slip events. In the model, contact elements are either in static contact or in the process of slipping to a new static contact. If stick-slip events are uncorrelated, the model further predicted that contact forces should increase to a critical velocity (V*) and then decrease at velocities greater than V*. We hypothesized that, like natural gecko setae, but unlike any conventional adhesive, gecko-like synthetic adhesives (GSAs) could adhere while sliding. To test the generality of our results and the validity of our model, we fabricated a GSA using a hard silicone polymer. While sliding, the GSA exhibited steady-state adhesion and velocity dependence similar to that of gecko setae. Observations at the interface indicated that macroscopically smooth sliding of the GSA emerged from randomly occurring stick-slip events in the population of flexible fibrils, confirming our model predictions.

Changing incidence of methicillin-resistant staphylococcus aureus skin abscesses in a pediatric emergency department.

OBJECTIVES: The primary objective of this study was to determine the etiology of skin abscesses in a pediatric emergency department (ED) during a 4-year period and to determine whether the incidence of methicillin-resistant Staphylococcus aureus (MRSA) skin abscesses has increased. The secondary objective was to characterize MRSA infections by antibiotic susceptibility during the same period. METHODS: A retrospective chart review examining all cases of skin abscess requiring an incision and drainage seen in a free-standing children's hospital ED from January 1, 2003, to December 30, 2006, was performed. Demographic and clinical data were abstracted from the medical records. Culture results, including sensitivities to antibiotics, were obtained to identify how many of these patients had MRSA. RESULTS: The charts of 442 children were analyzed, and 274 (62%) had MRSA isolated during the entire study period. In 2003, 36.2% of all drained abscesses were caused by MRSA, and by 2006, this increased to 66.5% (P < 0.001). Methicillin-resistant S. aureus isolates were sensitive to trimethoprim-sulfamethoxazole in 99% of the cases, but the sensitivity of MRSA to clindamycin decreased from 96% in 2003 to 87% by 2006. All isolates remain sensitive to rifampin, vancomycin, and gentamicin. CONCLUSIONS: The incidence of MRSA skin abscesses has increased in the pediatric ED population and now accounts for greater than 50% of all abscesses. If antimicrobial therapy is indicated for the treatment of these abscesses, cultures should be obtained, and antibiotics should be chosen to provide MRSA coverage.

A microfabricated wedge-shaped adhesive array displaying gecko-like dynamic adhesion, directionality and long lifetime.

Gecko adhesion has become a paradigmatic example of bio-inspired engineering, yet among the many gecko-like synthetic adhesives (GSAs), truly gecko-like performance remains elusive. Many GSAs have previously demonstrated one or two features of the gecko adhesive. We present a new wedge-shaped GSA that exhibits several gecko-like properties simultaneously: directional features; zero force at detachment; high ratio of detachment force to preload force; non-adhesive default state; and the ability to maintain performance while sliding, even after thousands of cycles. Individual wedges independently detach and reattach during sliding, resulting in high levels of shear and normal adhesion during drag. This behaviour provides a non-catastrophic failure mechanism that is desirable for applications such as climbing robots where sudden contact failure would result in serious falls. The effects of scaling patch sizes up to tens of square centimetres are also presented and discussed. Patches of 1 cm(2) had an adhesive pressure of 5.1 kPa while simultaneously supporting 17.0 kPa of shear. After 30 000 attachment/detachment cycles, a patch retained 67 per cent of its initial adhesion and 76 per cent of its initial shear without cleaning. Square-based wedges of 20 mum and 50 mum are manufactured in a moulding process where moulds are fabricated using a dual-side, dual-angle lithography process on quartz wafers with SU-8 photoresist as the mould material and polydimethylsiloxane as the cast material.

Frictional and elastic energy in gecko adhesive detachment.

Geckos use millions of adhesive setae on their toes to climb vertical surfaces at speeds of over 1 m s(-1). Climbing presents a significant challenge for an adhesive since it requires both strong attachment and easy, rapid removal. Conventional pressure-sensitive adhesives are either strong and difficult to remove (e.g. duct tape) or weak and easy to remove (e.g. sticky notes). We discovered that the energy required to detach adhering tokay gecko setae (W(d)) is modulated by the angle (theta) of a linear path of detachment. Gecko setae resist detachment when dragged towards the animal during detachment (theta = 30 degrees ) requiring W(d) = 5.0+/-0.86(s.e.) J m(-2) to detach, largely due to frictional losses. This external frictional loss is analogous to viscous internal frictional losses during detachment of pressure-sensitive adhesives. We found that, remarkably, setae possess a built-in release mechanism. Setae acted as springs when loaded in tension during attachment and returned elastic energy when detached along the optimal path (theta=130 degrees ), resulting in W(d) = -0.8+/-0.12 J m(-2). The release of elastic energy from the setal shaft probably causes spontaneous release, suggesting that curved shafts may enable easy detachment in natural, and synthetic, gecko adhesives.