Macrophages in wound healing: activation and plasticity.

Macrophages are critically involved in wound healing, from dampening inflammation to clearing cell debris and coordinating tissue repair. Within the wound, the complexity of macrophage function is increasingly recognized, with adverse outcomes when macrophages are inappropriately activated, such as in fibrosis or chronic non-healing wounds. Recent advances in in vivo and translational wound models, macrophage-specific deletions and new technologies to distinguish macrophage subsets, have uncovered the vast spectrum of macrophage activation and effector functions. Here, we summarize the main players in wound-healing macrophage activation and function, including cytokines, apoptotic cells, nucleotides and mechanical stimuli. We highlight recent studies demonstrating cooperation between these factors for optimal wound healing. Next, we describe recent technologies such as cell tracking and single-cell RNA-seq, which have uncovered remarkable plasticity and heterogeneity in blood-derived or tissue-resident macrophages and discuss the implications for wound healing. Lastly, we evaluate macrophage dysfunction in aberrant wound healing that occurs in aging, diabetes and fibrosis. A better understanding of the longevity and plasticity of wound-healing macrophages, and identification of unique macrophage subsets or specific effector molecules in wound healing, would shed light on the therapeutic potential of manipulating macrophage function for optimal wound healing

Evaluation for food packaging potential and environmental compatibility of novel degradable starch-polyethylene plastics

Linear low-density polyethylene films were prepared that contained native corn starch (7, 14, or 28%), low- or high-molecular weight oxidized- polyethylene (15%) and a pro-oxidant mixture (manganese 55 ppm and vegetable oil). Each plastic blend was first mixed in a twin-screw extruder at 195°C, pelletized, then cast into films using a single-screw extruder at 205°C. Oxidized-polyethylene addition to starch-polyethylene films did not impair transparency and thickness of the films, and relatively small reductions in film mechanical properties were observed;Thermal and photodegradation properties of each film were evaluated by 70°C forced-air oven treatment (20 days), by high-temperature high-humidity treatment in a steam chamber (20 days), and by exposure to ultraviolet light (365 nm) (4 weeks). Changes were determined in the mechanical properties, carbonyl index and molecular weight distribution of the films. The addition of oxidized-polyethylene, especially high-molecular weight oxidized-polyethylene, and up to 14% starch to the films, significantly increased the rate of thermal and photodegradation;Biodegradability was determined by using a pure-culture assay with ligninolytic Streptomyces badius 252, S. setonii 75Vi2, and S. viridosporus T7A and by using an extracellular-enzyme assay prepared from S. setonii 75Vi2 culture concentrate. The results from the pure-culture assay were inconclusive because of biomass accumulation on the film surface which inhibited chemical and biological degradation of the films. The extracellular-enzyme assay illustrated biodegradation for all 14 and 28% starch-polyethylene films, which suggests that the starch-polyethylene films could degrade in a biologically active compost environment;The food packaging potential of each film was evaluated. The films were stable in paraffin oil and alkali but were not stable in strong acids. Starch in the films did not impair the heat sealing property. Water vapor transmission of the films increased with increasing starch content, whereas oxygen permeability was not affected by the addition of oxidized-polyethylene nor starch. Starch in the films did not accelerate microbial growth in wrapped ground beef and most films showed relatively inert mechanical properties after use. Consequently, degradable films containing 7 to 14% starch, oxidized-polyethylene and pro-oxidant illustrated good degradability and food packaging potential

Integrated Systems Health Management (ISHM) Toolkit

A framework of software components has been implemented to facilitate the development of ISHM systems according to a methodology based on Reliability Centered Maintenance (RCM). This framework is collectively referred to as the Toolkit and was developed using General Atomics' Health MAP (TM) technology. The toolkit is intended to provide assistance to software developers of mission-critical system health monitoring applications in the specification, implementation, configuration, and deployment of such applications. In addition to software tools designed to facilitate these objectives, the toolkit also provides direction to software developers in accordance with an ISHM specification and development methodology. The development tools are based on an RCM approach for the development of ISHM systems. This approach focuses on defining, detecting, and predicting the likelihood of system functional failures and their undesirable consequences

Mutagenesis of Trichoderma reesei endoglucanase I: impact of expression host on activity and stability at elevated temperatures.

BackgroundTrichoderma reesei is a key cellulase source for economically saccharifying cellulosic biomass for the production of biofuels. Lignocellulose hydrolysis at temperatures above the optimum temperature of T. reesei cellulases (~50°C) could provide many significant advantages, including reduced viscosity at high-solids loadings, lower risk of microbial contamination during saccharification, greater compatibility with high-temperature biomass pretreatment, and faster rates of hydrolysis. These potential advantages motivate efforts to engineer T. reesei cellulases that can hydrolyze lignocellulose at temperatures ranging from 60-70°C.ResultsA B-factor guided approach for improving thermostability was used to engineer variants of endoglucanase I (Cel7B) from T. reesei (TrEGI) that are able to hydrolyze cellulosic substrates more rapidly than the recombinant wild-type TrEGI at temperatures ranging from 50-70°C. When expressed in T. reesei, TrEGI variant G230A/D113S/D115T (G230A/D113S/D115T Tr_TrEGI) had a higher apparent melting temperature (3°C increase in Tm) and improved half-life at 60°C (t1/2 = 161 hr) than the recombinant (T. reesei host) wild-type TrEGI (t1/2 = 74 hr at 60°C, Tr_TrEGI). Furthermore, G230A/D113S/D115T Tr_TrEGI showed 2-fold improved activity compared to Tr_TrEGI at 65°C on solid cellulosic substrates, and was as efficient in hydrolyzing cellulose at 60°C as Tr_TrEGI was at 50°C. The activities and stabilities of the recombinant TrEGI enzymes followed similar trends but differed significantly in magnitude depending on the expression host (Escherichia coli cell-free, Saccharomyces cerevisiae, Neurospora crassa, or T. reesei). Compared to N.crassa-expressed TrEGI, S. cerevisiae-expressed TrEGI showed inferior activity and stability, which was attributed to the lack of cyclization of the N-terminal glutamine in Sc_TrEGI and not to differences in glycosylation. N-terminal pyroglutamate formation in TrEGI expressed in S. cerevisiae was found to be essential in elevating its activity and stability to levels similar to the T. reesei or N. crassa-expressed enzyme, highlighting the importance of this ubiquitous modification in GH7 enzymes.ConclusionStructure-guided evolution of T. reesei EGI was used to engineer enzymes with increased thermal stability and activity on solid cellulosic substrates. Production of TrEGI enzymes in four hosts highlighted the impact of the expression host and the role of N-terminal pyroglutamate formation on the activity and stability of TrEGI enzymes

Bone Marrow-Derived Multipotent Stromal Cells Attenuate Inflammation in Obliterative Airway Disease in Mouse Tracheal Allografts

Obliterative bronchiolitis (OB) remains the most significant cause of death in long-term survival of lung transplantation. Using an established murine heterotopic tracheal allograft model, the effects of different routes of administration of bone marrow-derived multipotent stromal cells (MSCs) on the development of OB were evaluated. Tracheas from BALB/c mice were implanted into the subcutaneous tissue of major histocompatibility complex- (MHC-) disparate C57BL/6 mice. At the time of transplant, bone marrow-derived MSCs were administered either systemically or locally or via a combination of the two routes. The allografts were explanted at various time points after transplantation and were evaluated for epithelial integrity, inflammatory cell infiltration, fibrosis, and luminal obliteration. We found that the most effective route of bone marrow-derived MSC administration is the combination of systemic and local delivery. Treatment of recipient mice with MSCs suppressed neutrophil, macrophage, and T-cell infiltration and reduced fibrosis. These beneficial effects were observed despite lack of significant MSC epithelial engraftment or new epithelial cell generation. Our study suggests that optimal combination of systemic and local delivery of MSCs may ameliorate the development of obliterative airway disease through modulation of immune response

Photoluminescence of short-period GaAs/AlAs superlattices: A hydrostatic pressure and temperature study

URL:http://link.aps.org/doi/10.1103/PhysRevB.58.7222 DOI:10.1103/PhysRevB.58.7222The temperature and pressure dependence of type-I and -II transitions from photoluminescence (PL) spectra in a series of (GaAs)m/(AlAs)m superlattices show that the temperature dependence of energy bands can be described very well with a Bose-Einstein-type equation. From these measurements the parameters that describe the temperature dependence of excitonic transition energies and the corresponding broadening of the PL line are deduced. The pressure dependence of the PL linewidths of the type-I exciton as a function of pressure and temperature yield the intervalley deformation potential. Beyond the type-I-type-II crossover, the PL linewidth increases as a function of both pressure and temperature. The electron-phonon deformation potential for Γ-X scattering is found to be temperature dependent.We thank S. Satpathy and S. Zollner for valuable discussions. One of us ~H.R.C.! acknowledges support by the NSF under Grant No. DMR-9633107. M.C. thanks the U.S. Army for support through Grant No. DAAL03-92-0381. The work at Purdue University was supported by the National Science Foundation: Materials Research Science and Engineering Center Grant Nos. DMR 94-00415 and DMR 93-03186

The student-athletes coping strategy and sources of stress

The purpose of this study was to examine the sources of stress and coping strategies among university athletes. Fifteen athletes from a selected university Rugby Sport of Excellence Program in Malaysia aged between18 to 23 years old agreed to participate in this study. Starting from the top level, only 7% of the players have the experience playing at national level, a majority of 60% of them have the experience playing at the state level, 20% at the district level while 13% of the players only play at school level. Qualitative methodology was utilized in this investigation and the interview transcript was analyzed inductively. The results show four sources of stress: feeling nervous before the game, injuries, disruption from the audience and personal problems. Whereas, there are five coping strategies: social support, positive self-talk, mind set, remembering previous success and physical preparation. Recommendations were also suggested in the paper

Next-Generation Sequencing of HIV-1 Single Genome Amplicons

The analysis of HIV-1 sequences has helped understand the viral molecular epidemiology, monitor the development of antiretroviral drug resistance, and design candidate vaccines. The introduction of single genome amplification (SGA) has been a major advancement in the field, allowing for the characterization of multiple sequences per patient while preserving linkage among polymorphisms in the same viral genome copy. Sequencing of SGA amplicons is performed by capillary Sanger sequencing, which presents low throughput, requires a high amount of template, and is highly sensitive to template/primer mismatching. In order to meet the increasing demand for HIV-1 SGA amplicon sequencing, we have developed a platform based on benchtop next-generation sequencing (NGS) (IonTorrent) accompanied by a bioinformatics pipeline capable of running on computer resources commonly available at research laboratories. During assay validation, the NGS-based sequencing of 10 HIV-1 env SGA amplicons was fully concordant with Sanger sequencing. The field test was conducted on plasma samples from 10 US Navy and Marine service members with recent HIV-1 infection (sampling interval: 2005-2010; plasma viral load: 5,884-194,984 copies/ml). The NGS analysis of 101 SGA amplicons (median: 10 amplicons/individual) showed within-individual viral sequence profiles expected in individuals at this disease stage, including individuals with highly homogeneous quasispecies, individuals with two highly homogeneous viral lineages, and individuals with heterogeneous viral populations. In a scalability assessment using the Ion Chef automated system, 41/43 tested env SGA amplicons (95%) multiplexed on a single Ion 318 chip showed consistent gene-wide coverage \u3e50×. With lower sample requirements and higher throughput, this approach is suitable to support the increasing demand for high-quality and cost-effective HIV-1 sequences in fields such as molecular epidemiology, and development of preventive and therapeutic strategies

Free Fatty Acids Rewire Cancer Metabolism in Obesity-Associated Breast Cancer via Estrogen Receptor and mTOR Signaling

Obesity is a risk factor for postmenopausal estrogen receptor alpha (ERα)-positive (ER+) breast cancer. Molecular mechanisms underlying factors from plasma that contribute to this risk and how these mechanisms affect ERα signaling have yet to be elucidated. To identify such mechanisms, we performed whole metabolite and protein profiling in plasma samples from women at high risk for breast cancer, which led us to focus on factors that were differentially present in plasma of obese versus nonobese postmenopausal women. These studies, combined with in vitro assays, identified free fatty acids (FFA) as circulating plasma factors that correlated with increased proliferation and aggressiveness in ER+ breast cancer cells. FFAs activated both the ERα and mTOR pathways and rewired metabolism in breast cancer cells. Pathway preferential estrogen-1 (PaPE-1), which targets ERα and mTOR signaling, was able to block changes induced by FFA and was more effective in the presence of FFA. Collectively, these data suggest a role for obesity-associated gene and metabolic rewiring in providing new targetable vulnerabilities for ER+ breast cancer in postmenopausal women. Furthermore, they provide a basis for preclinical and clinical trials where the impact of agents that target ERα and mTOR signaling cross-talk would be tested to prevent ER+ breast cancers in obese postmenopausal women