3D printed structures for modeling the Young's modulus of bamboo parenchyma.

Bamboo is a sustainable, lightweight material that is widely used in structural applications. To fully develop micromechanical models for plants, such as bamboo, the mechanical properties of each individual type of tissue are needed. However, separating individual tissues and testing them mechanically is challenging. Here, we report an alternative approach in which micro X-ray computed tomography (µ-CT) is used to image moso bamboo (Phyllostachys pubescens). The acquired images, which correspond to the 3D structure of the parenchyma, are then transformed into physical, albeit larger scale, structures by 3D printing, and their mechanical properties are characterized. The normalized longitudinal Young's moduli of the fabricated structures depend on relative density raised to a power between 2 and 3, suggesting that elastic deformation of the parenchyma cellular structure involves considerable cell wall bending. The mechanical behavior of other biological tissues may also be elucidated using this approach. STATEMENT OF SIGNIFICANCE: Bamboo is a lightweight, sustainable engineering material widely used in structural applications. By combining micro X-ray computed tomography and 3D printing, we have produced bamboo parenchyma mimics and characterized their stiffness. Using this approach, we gained insight into bamboo parenchyma tissue mechanics, specifically the cellular geometry's role in longitudinal elasticity.

Tactic-specific benefits of polyandry in Chinook salmon Oncorhynchus tshawytscha.

This study examined whether polyandrous female Chinook salmon Oncorhynchus tshawytscha obtain benefits compared with monandrous females through an increase in hatching success. Both of the alternative reproductive tactics present in male O. tshawytscha (large hooknoses and small, precocious jacks) were used, such that eggs were either fertilized by a single male (from each tactic) or multiple males (using two males from the same or different tactics). The results show that fertilized eggs from the polyandrous treatments had a significantly higher hatching success than those from the monandrous treatments. It is also shown that sperm speed was positively related with offspring hatching success. Finally, there were tactic-specific effects on the benefits females received. The inclusion of jacks in any cross resulted in offspring with higher hatching success, with the cross that involved a male from each tactic providing offspring with the highest hatching success than any other cross. This study has important implications for the evolution of multiple mating and why it is so prevalent across taxa, while also providing knowledge on the evolution of mating systems, specifically those with alternative reproductive tactics.

Evaluation of the Pulmonary Toxicity of Ambient Particulate Matter From Camp Victory, Iraq.

Anecdotal reports in the press and epidemiological studies suggest that deployment to Iraq and Afghanistan may be associated with respiratory diseases and symptoms in U.S. military personnel and veterans. Exposures during military operations were complex, but virtually all service members were exposed to high levels of respirable, geogenic dust. Inhalation of other dusts has been shown to be associated with adverse health effects, but the pulmonary toxicity of ambient dust from Iraq has not been previously studied. The relative toxicity of Camp Victory dust was evaluated by comparing it to particulate matter from northern Kuwait, a standard U.S. urban dust, and crystalline silica using a single intratracheal instillation in rats. Lung histology, protein levels, and cell counts were evaluated in the bronchoalveolar lavage fluid 1-150 d later. The Iraq dust provoked an early significant, acute inflammatory response. However, the level of inflammation in response to the Iraq dust, U.S. urban dust, and Kuwait dust rapidly declined and was nearly at control levels by the end of the study At later times, animals exposed to the Iraq, U.S. urban, or Kuwait dusts showed increased small airway remodeling and emphysema compared to silica-exposed and control animals without evidence of fibrosis or premalignant changes. The severity and persistence of pulmonary toxicity of these three dusts from the Middle East resemble those of a U.S. urban dust and are less than those of silica. Therefore, Iraq dust exposure is not highly toxic, but similar to other poorly soluble low-toxicity dusts.

Chromosomal ampC mutations in cefpodoxime-resistant, ESBL-negative uropathogenic Escherichia coli.

AmpC ß-lactamase is an enzyme commonly produced by Escherichia coli that causes resistance to cephalosporins and penicillins. Enzyme production is controlled by the strength of the promoter encoded by the chromosomal ampC gene, with the level of production affected by the presence of certain mutations in this region. This study sets out to determine the prevalence of ampC promoter mutations present in a group of uropathogenic E. coli strains. A total of 50 clinical strains of E. coli were collected from urine samples between June 2011 and November 2011. Strains were investigated for the presence of mutations in the chromosomal ampC promoter region by amplification and sequencing of a 271 bp product. The presence of ampC-carrying plasmids derived from other species was also determined, to exclude these from further analysis. ampC-carrying plasmids were found in 10 of the 50 strains, all of which were of the CIT-type. Analysis of the chromosomal ampC promoter region in the 40 remaining strains showed mutations at 16 different positions, with 18 different genotype patterns detected overall. The most common ampC chromosomal mutation, present in 25 of 40 strains, was a T --> A transition at position -32. This mutation has been shown by others to increase enzyme production by up to 46-fold. Altogether, three separate mutations (-32, -42 and -13ins) were present in 90% of the 40 non-plasmid strains, indicating a strong association with the resistance observed. It appears, therefore, that the majority of AmpC-mediated resistance in E. coli can be accounted for by just three point mutations in the chromosome.

The de morton mobility index: normative data for a clinically useful mobility instrument.

Determining mobility status is an important component of any health assessment for older adults. In order for a mobility measure to be relevant and meaningful, normative data are required for comparison to a healthy reference population. The DEMMI is the first mobility instrument to measure mobility across the spectrum from bed bound to functional levels of independent mobility. In this cross-sectional observational study, normative data were obtained for the DEMMI from a population of 183 healthy, community-dwelling adults age 60+ who resided in Vancouver, Canada and Melbourne, Australia. Older age categories had significantly lower DEMMI mobility mean scores (P < 0.05), as did individuals who walked with a mobility aid or lived in semi-independent living (assisted living or retirement village), whereas DEMMI scores did not differ by sex (P = 0.49) or reported falls history (P = 0.21). Normative data for the DEMMI mobility instrument provides vital reference scores to facilitate its use across the mobility spectrum in clinical, research, and policymaking settings.

Neural correlates of attentional expertise in long-term meditation practitioners.

Meditation refers to a family of mental training practices that are designed to familiarize the practitioner with specific types of mental processes. One of the most basic forms of meditation is concentration meditation, in which sustained attention is focused on an object such as a small visual stimulus or the breath. In age-matched participants, using functional MRI, we found that activation in a network of brain regions typically involved in sustained attention showed an inverted u-shaped curve in which expert meditators (EMs) with an average of 19,000 h of practice had more activation than novices, but EMs with an average of 44,000 h had less activation. In response to distracter sounds used to probe the meditation, EMs vs. novices had less brain activation in regions related to discursive thoughts and emotions and more activation in regions related to response inhibition and attention. Correlation with hours of practice suggests possible plasticity in these mechanisms.

Approaches to the release of a master cell bank of PER.C6 cells; a novel cell substrate for the manufacture of human vaccines.

At Merck and Co. we have developed a recombinant E1 deficient adenovirus type 5 vaccine vector for HIV-1 and have adopted the PER.C6 cell line as a cell substrate for the manufacture of this vector for Phase I and II clinical trials. The PER.C6 cell line was developed at Crucell by the transfection of human primary embryonic retinoblasts with a transgene of E1 constructed with a minimum of E1 coding sequences to preclude homologous recombination generating replication-competent adenovirus, between E1 sequences in PER.C6 and adenovirus vectors with E1 deletions of the same molecular coordinates. We have developed a Master Cell Bank (MCB) of PER.C6 cells under serum-free conditions of suspension culture from a vial of PER.C6 cells obtained from Crucell. This MCB has been released according to an extensive panel of testing for the detection of adventitious viral agents, including assays for sterility and mycoplasma, in vivo and in vitro assays for the detection of viruses of human, bovine and porcine origin, replication competent adenoviruses, sensitive PERT assays for the detection of RT in supernatants of co-cultivations, electron microscopy and a panel of PCR-based assays for specific human and animal viruses. This MCB has been used for the manufacture of vaccine vector supporting a number of IND submissions for Phase I clinical trials over a three-year period during which the panel of PCR testing applied to the MCB has been judiciously expanded. Advances in QPCR technology, liquid handling systems, and more recently mass spectrometry offer the possibility that very broad panels of primers and probes capable of the detection of all known human viruses can be applied routinely to support the release of biologicals for human clinical trials. The impact of this breadth of testing on the continued reliance of classical in vivo and in vitro assays for adventitious viruses is clearly an emerging issue worthy of serious debate.

Tumorigenicity assessments of Per.C6 cells and of an Ad5-vectored HIV-1 vaccine produced on this continuous cell line.

PER.C6, a cell line derived from human embryonic retinal cells transformed with the Adenovirus Type 5 (Ad5) E1A and E1B genes, is used to produce E1-deleted Ad5 vectors such as the MRKAd5 HIV-1 gag vaccine. While whole, live PER.C6 cells are capable of growing as tumours when transplanted subcutaneously into immunodeficient nude mice at a high dosage, the process for vaccine production includes filtration steps and other methods which effectively preclude contamination by intact viable substrate cells. However, because of the neoplastic nature of this cell line, we carried out a series of investigations to assess the tumorigenic risk posed by residuals from the cell substrate in a vaccine. To address concerns about transmission of oncogenic DNA, we demonstrated that purified PER.C6 cellular DNA does not induce tumours in newborn hamsters or nude mice. To address concerns about other potential residuals, including hypothetical adventitious tumour viruses, we demonstrated that a PER.C6 cell lysate and a MRKAd5 HIV-1 gag vaccine produced on PER.C6 cells do not induce tumours in newborn hamsters or newborn rats. These results, in conjunction with the wide panel of viral safety tests performed on these cells, support the safety of the PER.C6 as a cell substrate for vaccine production.

Clinical efficacy assessment in photodamaged skin of 0.5% and 1.0% idebenone.

Idebenone is an antioxidant lower molecular weight analogue of coenzyme Q10. Previously, idebenone was shown to be a very effective antioxidant in its ability to protect against cell damage from oxidative stress in a variety of biochemical, cell biological, and in vivo methods, including its ability to suppress sunburn cell (SBC) formation in living skin. However, no clinical studies have been previously conducted to establish the efficacy of idebenone in a topical skincare formulation for the treatment of photodamaged skin. In this nonvehicle control study, 0.5% and 1.0% idebenone commercial formulations were evaluated in a clinical trial for topical safety and efficacy in photodamaged skin. Forty-one female subjects, aged 30-65, with moderate photodamaged skin were randomized to use a blind labelled (either 0.5% or 1.0% idebenone in otherwise identical lotion bases) skincare preparation twice daily for six weeks. Blinded expert grader assessments for skin roughness/dryness, fine lines/wrinkles, and global improvement in photodamage were performed at baseline, three weeks and six weeks. Electrical conductance readings for skin surface hydration and 35 mm digital photography were made at baseline after six weeks. Punch biopsies were taken from randomly selected subjects, baseline and after six weeks, and stained for certain antibodies (interleukin IL-6, interleukin IL-1b, matrixmetalloproteinase MMP-1, collagen I) using immunofluorescence microscopy. After six weeks' use of the 1.0% idebenone formula, a 26% reduction in skin roughness/dryness was observed, a 37% increase in skin hydration, a 29% reduction in fine lines/wrinkles, and a 33% improvement in overall global assessment of photodamaged skin. For the 0.5% idebenone formulation, a 23% reduction in skin roughness/dryness was observed, a 37% increase in skin hydration, a 27% reduction in fine lines/wrinkles, and a 30% improvement in overall global assessment of photodamaged skin. The immunofluorescence staining revealed a decrease in IL-1b, IL-6, and MMP-1 and an increase in collagen I for both concentrations.

Idebenone: a new antioxidant - Part I. Relative assessment of oxidative stress protection capacity compared to commonly known antioxidants.

Topical applications of skin care products containing antioxidants have become increasingly popular. Numerous studies have elucidated the biological effects of these substances. General antiaging effects, anti-inflammatory properties, photoprotective properties, and prevention of ultraviolet (UV) immunosuppression have been documented. However, a standardized method to characterize and compare the properties and oxidative stress protection capacity of antioxidants was lacking. A multistep in vitro process utilizing a variety of biochemical and cell biological methods combined with in vivo studies was designed to compare the oxidative stress protective capacity of commonly used antioxidants. Data were presented for L-ascorbic acid, dl-alpha-tocopherol, kinetin, dl-alpha lipoic acid, ubiquinone, and idebenone. Methods included using UV-induced radical trapping/scavenging capacity measured by photochemiluminescence, pro-oxidative systems (LDL-CuSO(4), microsome-NADPH/ADP/Fe(3+)) with measurement of primary and secondary oxidation products, UVB irradiation of human keratinocytes, and in vivo evaluation, using the human sunburn cell (SBC) assay. Correlation and trends between in vitro and in vivo results were established, and the standardized test protocol was used to quantify oxidative stress protection capacity of antioxidants. Summarizing and totaling the data equally weighted for each oxidative stress study, the overall oxidative protection capacity scores of 95, 80, 68, 55, 52, and 41 were obtained for idebenone, dl-alpha tocopherol, kinetin, ubiquinone, L-ascorbic acid, and dl-alpha lipoic acid, respectively. The higher the score, the more effective the overall oxidative stress protection capacity of the antioxidant became. This multistep protocol may serve as a standard in investigating and comparing new putative antioxidants for topical use as well as a valuable tool to assess the anti-inflammatory properties, photoprotective properties, and prevention of UV immunosuppression of topical antioxidants.

An evaluation of the efficacy of Aqualox for microbiological control of industrial cooling tower systems.

A comprehensive sampling protocol was employed to evaluate the efficacy of Aqualox, a biocide based on electrochemically activated water, against legionellae and heterotrophic bacteria in two industrial cooling tower systems. Both of the towers in the study remained free from evidence of Legionella spp. contamination throughout a five-month evaluation period, despite the previously demonstrated presence of legionellae in one of the test towers, and in two other towers on the same site, at levels well in excess of UK Health and Safety Commission (HSC) Approved Code of Practice and Guidance (ACOP) upper action limits. Levels of heterotrophic bacteria were controlled below 10(4) cfu/mL in both towers throughout most of the trial. Results also provided indirect evidence of significant activity against biofilm bacteria, with biofilm removal beginning almost immediately after commissioning of the Aqualox treatment systems. The results were particularly encouraging as the two towers studied had a long history of poor microbiological control using conventional bromine-based biocide products. Significant differences were observed between laboratory measurements of total viable counts on frequent liquid samples and those obtained from dip slides following HSC recommendations.

The human genome and public policy: a nursing perspective.

The Human Genome Project has completed a rough draft of the sequence that comprises human DNA. For the first time, the scientific discoveries have been conducted in tandem with research exploring the social, legal, and ethical implications of the research. Several committees have studied the policy implications of the genetics information explosion. The Secretary's Advisory Committee on Genetic Testing's report was received by the Secretary of the U.S. Department of Health and Human Services in November 2000 and contains recommendations for the oversight of genetic testing. Public policy issues that affect nursing practice in the area of genetics must be explored by individual nurses and professional nursing organizations.

Social support, intrusive thoughts, and quality of life in breast cancer survivors.

This study explores the moderating effect of social support on the relationship between cancer-related intrusive thoughts and quality of life. Sixty-four breast cancer survivors completed self-report measures of appraisal social support (the disclosure of thoughts and feelings to significant others), cancer-related intrusive thoughts, and quality of life. Controlling for demographic and treatment variables, the negative impact of cancer-related intrusive thoughts on both physical and mental quality of life measures was moderated by appraisal social support. For women with high levels of appraisal support, cancer-related intrusive thoughts had no significant relationship with quality of life. However, for women with low levels of appraisal support, the relationship between cancer-related intrusive thoughts and quality of life was significant and negative. These results suggest that appraisal social support can mitigate the impact of traumatic life events.

Nanoparticle halos: a new colloid stabilization mechanism.

A new mechanism for regulating the stability of colloidal particles has been discovered. Negligibly charged colloidal microspheres, which flocculate when suspended alone in aqueous solution, undergo a remarkable stabilizing transition upon the addition of a critical volume fraction of highly charged nanoparticle species. Zeta potential analysis revealed that these microspheres exhibited an effective charge buildup in the presence of such species. Scanning angle reflectometry measurements indicated, however, that these nanoparticle species did not adsorb on the microspheres under the experimental conditions of interest. It is therefore proposed that highly charged nanoparticles segregate to regions near negligibly charged microspheres because of their repulsive Coulombic interactions in solution. This type of nanoparticle haloing provides a previously unreported method for tailoring the behavior of complex fluids.

Control of directionality in integrase-mediated recombination: examination of recombination directionality factors (RDFs) including Xis and Cox proteins.

Similarity between the DNA substrates and products of integrase-mediated site-specific recombination reactions results in a single recombinase enzyme being able to catalyze both the integration and excision reactions. The control of directionality in these reactions is achieved through a class of small accessory factors that favor one reaction while interfering with the other. These proteins, which we will refer to collectively as recombination directionality factors (RDFs), play architectural roles in reactions catalyzed by their cognate recombinases and have been identified in conjunction with both tyrosine and serine integrases. Previously identified RDFs are typically small, basic and have diverse amino acid sequences. A subset of RDFs, the cox genes, also function as transcriptional regulators. We present here a compilation of all the known RDF proteins as well as those identified through database mining that we predict to be involved in conferring recombination directionality. Analysis of this group of proteins shows that they can be grouped into distinct sub-groups based on their sequence similarities and that they are likely to have arisen from several independent evolutionary lineages. This compilation will prove useful in recognizing new proteins that confer directionality upon site-specific recombination reactions encoded by plasmids, transposons, phages and prophages.

Vaccinia virus blocks gamma interferon signal transduction: viral VH1 phosphatase reverses Stat1 activation.

We have analyzed the effects of vaccinia virus (VV) on gamma interferon (IFN-gamma) signal transduction. Infection of cells with VV 1 to 2 h prior to treatment with IFN-gamma inhibits phosphorylation and nuclear translocation of Stat1 and consequently blocks accumulation of mRNAs normally induced by IFN-gamma. While phosphorylation of other proteins in the IFN-gamma pathway was not affected, activation of Stat1 by other ligand-receptor systems was also blocked by VV. This block of Stat1 activation was dose dependent, and although viral protein synthesis was not required, entry and uncoating of viral cores appear to be needed to block the accumulation of phosphorylated Stat1. These results suggest that a virion component is responsible for the effect. VV virions contain a phosphatase (VH1) that is sensitive to the phosphatase inhibitor Na(3)VO(4) but not to okadaic acid. Addition of Na(3)VO(4) but not okadaic acid restored normal Stat1 phosphorylation levels in VV-infected cells. Moreover, virions containing reduced levels of VH1 were unable to block the IFN-gamma signaling pathway. In vitro studies show that the phosphatase can bind and dephosphorylate Stat1, indicating that this transcription factor can be a substrate for VH1. Our results reveal a novel mechanism by which VV interferes with the onset of host immune responses by blocking the IFN-gamma signal cascade through the dephosphorylating activity of the viral phosphatase VH1.

Modulation of the multidrug-resistance phenotype by new tropane alkaloid aromatic esters from Erythroxylum pervillei.

Nine tropane alkaloid aromatic esters (1-9) were isolated from the roots of Erythroxylum pervillei by following their potential to reverse multidrug-resistance with vinblastine-resistant oral epidermoid carcinoma (KB-V1) cells. All isolates, including seven new structures (3-9), were evaluated against a panel of human cancer cell lines, and it was found that alkaloids 3 and 5-9 showed the greatest activity with KB-V1 cells assessed in the presence of vinblastine, suggesting that these new compounds are potent modulators of P-glycoprotein. Confirmatory results were obtained with human ovarian adenocarcinoma (SKVLB) cells evaluated in the presence of adriamycin and synergistic studies performed with several cell lines from the NCI tumor panel. The structures of the new compounds were determined using spectroscopic techniques. Single-crystal X-ray analysis was performed on the monoester, tropane-3 alpha,6 beta,7 beta-triol 3-phenylacetate (1).