Scaling of claw sharpness: mechanical constraints reduce attachment performance in larger insects.
Claws are the most widespread attachment devices in animals, but comparatively little is known about the mechanics of claw attachment. A key morphological parameter in determining attachment ability is claw sharpness; however, there is a conflict between sharpness and fracture resistance. Sharper claws can interlock on more surfaces but are more likely to break. Body size interacts with this conflict such that larger animals should have much blunter claws and consequently poorer attachment ability than smaller animals. This expected size-induced reduction in attachment performance has not previously been investigated, and it is unclear how animals deal with this effect, and whether it indeed exists. We explored the scaling of claw sharpness with body size using four insect species (Nauphoeta cinerea, Gromphadorhina portentosa, Atta cephalotes and Carausius morosus) each covering a large size range. The scaling of claw sharpness varied significantly between species, suggesting that they face different pressures regarding claw function. Attachment forces were measured for A. cephalotes and G. portentosa (which had different scaling of claw sharpness) on several rough surfaces using a centrifuge setup. As expected, attachment performance was poorer in larger animals. Firstly, larger animals were more likely to slip, although this effect depended on the scaling of claw sharpness. Secondly, when they gripped, they attached with smaller forces relative to their weight. This size-induced reduction in attachment performance has significant implications for the attachment ability of larger animals on rough surfaces.


IL-20 is epigenetically regulated in NSCLC and down regulates the expression of VEGF.
IL-20 is a pleiotrophic member of the IL-10 family and plays a role in skin biology and the development of haematopoietic cells. Recently, IL-20 has been demonstrated to have potential anti-angiogenic effects in non-small cell lung cancer (NSCLC) by down regulating COX-2. The expression of IL-20 and its cognate receptors (IL-20RA/B and IL-22R1) was examined in a series of resected fresh frozen NSCLC tumours. Additionally, the expression and epigenetic regulation of this family was examined in normal bronchial epithelial and NSCLC cell lines. Furthermore, the effect of IL-20 on VEGF family members was examined. The expression of IL-20 and its receptors are frequently dysregulated in NSCLC. IL-20RB mRNA was significantly elevated in NSCLC tumours (p<0.01). Protein levels of the receptors, IL-20RB and IL-22R1, were significantly increased (p<0.01) in the tumours of NSCLC patients. IL-20 and its receptors were found to be epigenetically regulated through histone post-translational modifications and DNA CpG residue methylation. In addition, treatment with recombinant IL-20 resulted in decreased expression of the VEGF family members at the mRNA level. This family of genes are dysregulated in NSCLC and are subject to epigenetic regulation. Whilst the anti-angiogenic properties of IL-20 require further clarification, targeting this family via epigenetic means may be a viable therapeutic option in lung cancer treatment.


Role of SUMOylation in full antiestrogenicity.
The selective estrogen receptor downregulator (SERD) fulvestrant can be used as second-line treatment for patients relapsing after treatment with tamoxifen, a selective estrogen receptor modulator (SERM). Unlike tamoxifen, SERDs are devoid of partial agonist activity. While the full antiestrogenicity of SERDs may result in part from their capacity to downregulate levels of estrogen receptor alpha (ERα) through proteasome-mediated degradation, SERDs are also fully antiestrogenic in the absence of increased receptor turnover in HepG2 cells. Here we report that SERDs induce the rapid and strong SUMOylation of ERα in ERα-positive and -negative cell lines, including HepG2 cells. Four sites of SUMOylation were identified by mass spectrometry analysis. In derivatives of the SERD ICI164,384, SUMOylation was dependent on the length of the side chain and correlated with full antiestrogenicity. Preventing SUMOylation by the overexpression of a SUMO-specific protease (SENP) deSUMOylase partially derepressed transcription in the presence of full antiestrogens in HepG2 cells without a corresponding increase in activity in the presence of agonists or of the SERM tamoxifen. Mutations increasing transcriptional activity in the presence of full antiestrogens reduced SUMOylation levels and suppressed stimulation by SENP1. Our results indicate that ERα SUMOylation contributes to full antiestrogenicity in the absence of accelerated receptor turnover.


Cerenkov Luminescence Imaging for Accurate Placement of Radioactive Plaques in Episcleral Brachytherapy of Intraocular Tumors.
The purpose of this study was to determine the feasibility of using Cerenkov luminescence imaging (CLI) to facilitate plaque placement during episcleral brachytherapy of intraocular tumors. Ruthenium-106 (Ru-106) decays to rhodium-106, which in turn emits high-energy beta particles. When the electrons propagate through the eyewall, the so-called Cerenkov effect leads to emission of weak light, which can be captured by high-sensitivity charge-couple device (CCD) cameras. Enucleated porcine eyes were prepared with tumor phantoms made of melanin-containing gelatin. The anterior portion of the globe was removed, and different Ru-106 plaque types (designated CCA, CCB, COB, and CIA) with activities ranging from 6.8 to 16.7 MBq were sutured to the sclera overlying the tumor phantom. The globe was placed in a transparent container with saline. CLI was performed through the anterior opening of the eye using a cooled electron-multiplying CCD camera. Exposure times between 5 and 60 seconds produced good quality images of the Cerenkov light. There was a linear relationship between plaque activity and Cerenkov radiance. The perimeters of the CCA and CCB plaques could be seen clearly as circles of light symmetrically surrounding the tumor phantoms. Notched COB and CIA plaques led to images revealing their actual positions in relation to the optic disc and ciliary body, respectively. Simulated plaque tilting resulted in diffuse demarcation of the light. The study indicates that CLI is a feasible method to ensure accurate placement of Ru-106 plaques in brachytherapy of intraocular tumors. CLI may offer a new tool to improve and document plaque placement, both perioperatively and postoperatively.


Local effects in the X-ray absorption spectrum of salt water.
Both first-principles molecular dynamics and theoretical X-ray absorption spectroscopy have been used to investigate the aqueous solvation of cations in 0.5 M MgCl(2), CaCl(2), and NaCl solutions. We focus here on the species-specific effects that Mg(2+), Ca(2+), and Na(+) have on the X-ray absorption spectrum of the respective solutions. For the divalent cations, we find that the hydrogen-bonding characteristics of the more rigid magnesium first-shell water molecules differ from those in the more flexible solvation shell surrounding calcium. In particular, the first solvation shell water molecules of calcium are able to form acceptor hydrogen bonds, and this results in an enhancement of a post-edge peak near 540 eV. The absence of acceptor hydrogen bonds for magnesium first shell water molecules provides an explanation for the experimental and theoretical observation of a lack of enhancement at the post-main-edge peak. For the sodium monovalent cation we find that the broad tilt angle distribution results in a broadening of postedge features, despite populations in donor-and-acceptor configurations consistent with calcium. We also present the reaveraged spectra of the MgCl(2), CaCl(2), and NaCl solutions and show that trends apparent with increasing concentration (0.5, 2.0, 4.0 M) are consistent with experiment. Finally, we examine more closely both the effect that cation coordination number has on the hydrogen-bonding network and the relative perturbation strength of the cations on lone pair oxygen orbitals.


Heterogeneity of action of mechanisms among antimycotic imidazoles.
The three imidazole antimycotics clotrimazole, miconazole, and ketoconazole all inhibit the demethylation of lanosterol to ergosterol, resulting in inhibition of growth of Saccharomyces cerevisiae; this is a fungistatic action. At higher concentrations clotrimazole and miconazole are fungicidal, whereas ketoconazole is not. The fungicidal action reflects direct membrane damage by the imidazoles. Evidence for this is that ketoconazole is markedly less active than the other imidazoles in its ability to allow methylene blue entry into cells and to disrupt liposome model membranes. The possible clinical significance of these findings is discussed.


Additional cash incentive within a conditional cash transfer scheme: a 'controlled before and during' design evaluation study from India.
Conditional Cash Transfer (CCT) schemes have shown largely favorable changes in the health seeking behavior. This evaluation study assesses the process and performance of an Additional Cash Incentive (ACI) scheme within an ongoing CCT scheme in India, and document lessons. A controlled before and during design study was conducted in Madhya Pradesh state of India, from August 2007 to March 2008, with increased in institutional deliveries as a primary outcome. In depth interviews, focus group discussions and household surveys were done for data collection. Lack of awareness about ACI scheme amongst general population and beneficiaries, cumbersome cash disbursement procedure, intricate eligibility criteria, extensive paper work, and insufficient focus on community involvement were the major implementation challenges. There were anecdotal reports of political interference and possible scope for corruption. At the end of implementation period, overall rate of institutional deliveries had increased in both target and control populations; however, the differences were not statistically significant. No cause and effect association could be proven by this study. Poor planning and coordination, and lack of public awareness about the scheme resulted in low utilization. Thus, proper IEC and training, detailed implementation plan, orientation training for implementer, sufficient budgetary allocation, and community participation should be an integral part for successful implementation of any such scheme. The lesson learned this evaluation study may be useful in any developing country setting and may be utilized for planning and implementation of any ACI scheme in future.


End-of-life decision making in Europe and Australia: a physician survey.
The frequencies with which physicians make different medical end-of-life decisions (ELDs) may differ between countries, but comparison between countries has been difficult owing to the use of dissimilar research methods. A written questionnaire was sent to a random sample of physicians from 9 specialties in 6 European countries and Australia to investigate possible differences in the frequencies of physicians' willingness to perform ELDs and to identify predicting factors. Response rates ranged from 39% to 68% (N = 10 139). Using hypothetical cases, physicians were asked whether they would (probably) make each of 4 ELDs. In all the countries, 75% to 99% of physicians would withhold chemotherapy or intensify symptom treatment at the request of a patient with terminal cancer. In most cases, more than half of all physicians would also be willing to deeply sedate such a patient until death. However, there was generally less willingness to administer drugs with the explicit intention of hastening death at the request of the patient. The most important predictor of ELDs was a request from a patient with decisional capacity (odds ratio, 2.1-140.0). Shorter patient life expectancy and uncontrollable pain were weaker predictors but were more stable across countries and across the various ELDs (odds ratios, 1.1-2.4 and 0.9-2.4, respectively). Cultural and legal factors seem to influence the frequencies of different ELDs and the strength of their determinants across countries, but they do not change the essence of decision making.


Comparative evaluation of efficacy of 0.2% Chlorhexidine, Listerine and Tulsi extract mouth rinses on salivary Streptococcus mutans count of high school children--RCT.
To assess and compare the effect of 0.2% Chlorhexidine mouth rinse, Listerine mouth rinse and 4% Tulsi extract mouth rinse on salivary Streptococcus mutans level. The present study is an experimental study of cross over type, employing Latin square design. 45 school children aged 14-15 years were divided into three groups/blocks. The baseline unstimulated saliva samples were obtained from each group and assessed for Streptococcus mutans counts. The study was divided in to three phases, each phase lasted for 8 days separated by a washout period of 15 days in between them. Groups A, B and C were treated with 0.2% Chlorhexidine, Listerine and 4% Tulsi extract mouth rinses respectively in the phase I. The study subjects were instructed to use the assigned mouth rinse twice daily for 1 min for 7 days. On day 8th the subjects were instructed to use the mouth rinse only once in the morning. The follow up unstimulated saliva samples were collected 1h after the use of the assigned mouth rinse and assessed for salivary Streptococcus mutans counts. After phase I, mouth rinses were crossed over as dictated by the Latin square design in phase II and III. All the three mouth rinses have individually shown a statistically significant reduction in the salivary Streptococcus mutans counts. When the three mouth rinses were compared the difference did not reach statistical significance. Tulsi has stood the test and is as effective as Chlorhexidine and Listerine in reducing the salivary S. mutans levels.


Yin and yang, or peas in a pod? Individual-sport versus team-sport athletes and altitude training.
The question of whether altitude training can enhance subsequent sea-level performance has been well investigated over many decades. However, research on this topic has focused on athletes from individual or endurance sports, with scant number of studies on team-sport athletes. Questions that need to be answered include whether this type of training may enhance team-sport athlete performance, when success in team-sport is often more based on technical and tactical ability rather than physical capacity per se. This review will contrast and compare athletes from two sports representative of endurance (cycling) and team-sports (soccer). Specifically, we draw on the respective competition schedules, physiological capacities, activity profiles and energetics of each sport to compare the similarities between athletes from these sports and discuss the relative merits of altitude training for these athletes. The application of conventional live-high, train-high; live-high, train-low; and intermittent hypoxic training for team-sport athletes in the context of the above will be presented. When the above points are considered, we will conclude that dependent on resources and training objectives, altitude training can be seen as an attractive proposition to enhance the physical performance of team-sport athletes without the need for an obvious increase in training load.


Natural Products and Chemical Biology Tools: Alternatives to Target Epigenetic Mechanisms in Cancers.
DNA methylation and histone acetylation are widely studied epigenetic modifications. They are involved in numerous pathologies such as cancer, neurological disease, inflammation, obesity, etc. Since the discovery of the epigenome, numerous compounds have been developed to reverse DNA methylation and histone acetylation aberrant profile in diseases. Among them several were inspired by Nature and have a great interest as therapeutic molecules. In the quest of finding new ways to target epigenetic mechanisms, the use of chemical tools is a powerful strategy to better understand epigenetic mechanisms in biological systems. In this review we will present natural products reported as DNMT or HDAC inhibitors for anticancer treatments. We will then discuss the use of chemical tools that have been used in order to explore the epigenome.


Intrinsic information carriers in combinatorial dynamical systems.
Many proteins are composed of structural and chemical features--"sites" for short--characterized by definite interaction capabilities, such as noncovalent binding or covalent modification of other proteins. This modularity allows for varying degrees of independence, as the behavior of a site might be controlled by the state of some but not all sites of the ambient protein. Independence quickly generates a startling combinatorial complexity that shapes most biological networks, such as mammalian signaling systems, and effectively prevents their study in terms of kinetic equations-unless the complexity is radically trimmed. Yet, if combinatorial complexity is key to the system's behavior, eliminating it will prevent, not facilitate, understanding. A more adequate representation of a combinatorial system is provided by a graph-based framework of rewrite rules where each rule specifies only the information that an interaction mechanism depends on. Unlike reactions, which deal with molecular species, rules deal with patterns, i.e., multisets of molecular species. Although the stochastic dynamics induced by a collection of rules on a mixture of molecules can be simulated, it appears useful to capture the system's average or deterministic behavior by means of differential equations. However, expansion of the rules into kinetic equations at the level of molecular species is not only impractical, but conceptually indefensible. If rules describe bona fide patterns of interaction, molecular species are unlikely to constitute appropriate units of dynamics. Rather, we must seek aggregate variables reflective of the causal structure laid down by the rules. We call these variables "fragments" and the process of identifying them "fragmentation." Ideally, fragments are aspects of the system's microscopic population that the set of rules can actually distinguish on average; in practice, it may only be feasible to identify an approximation to this. Most importantly, fragments are self-consistent descriptors of system dynamics in that their time-evolution is governed by a closed system of kinetic equations. Taken together, fragments are endogenous distinctions that matter for the dynamics of a system, which warrants viewing them as the carriers of information. Although fragments can be thought of as multisets of molecular species (an extensional view), their self-consistency suggests treating them as autonomous aspects cut off from their microscopic realization (an intensional view). Fragmentation is a seeded process that depends on the choice of observables whose dynamics one insists to describe. Different observables can cause distinct fragmentations, in effect altering the set of information carriers that govern the behavior of a system, even though nothing has changed in its microscopic constitution. In this contribution, we present a mathematical specification of fragments, but not an algorithmic implementation. We have described the latter elsewhere in rather technical terms that, although effective, were lacking an embedding into a more general conceptual framework, which we here provide.


Complication and Reoperation Rates Following Surgical Management of Cervical Spondylotic Myelopathy in Medicare Beneficiaries.
Retrospective database review. To compare complication and reoperation rates after anterior cervical discectomy and fusion (ACDF), posterior cervical fusion (PCFs), and anterior cervical corpectomy and fusion (ACCF) for cervical spondylotic myelopathy (CSM) using a large national database of Medicare beneficiaries. CSM is the most common cause of myelopathy in patients over 55 years and is considered the most common cause of spinal cord dysfunction in the world. Surgical treatment includes ACDF, PCF, or ACCF procedures. The PearlDiver database (2005-2012) was utilized to determine revision rates after surgical treatment of CSM by one of the aforementioned surgical treatments. Specifically, 1 to 2 level ACDF, ACCF, and PCF and 3+ level PCF cohorts were included. Each cohort was stratified by the age of 65 years. Survivorship curves were graphed and compared. Of the patients younger than 65 years of age, there were 10,557 patients treated with 1 to 2 level ACDF procedures, 1319 patients with 1 to 2 level PCF procedures, 1203 patients with 1 to 2 level ACCF procedures, and 2312 patients treated with 3+ level PCF procedures. Of the elderly patients, 24,310 patients were treated with 1 to 2 level ACDFs, 4776 with 1 to 2 level PCF procedures, 3109 with 1 to 2 level ACCFs, and 7760 with 3+ level PCFs. Patients younger than 65 years of age were significantly more likely to have a reoperation procedure, than those 65 years or older when analyzing ACCF, ACDF, and 3+ level PCF procedures. ACCFs were significantly more likely than ACDFs to require reoperation. Patients treated with PCF were consistently more likely to have nondysphagia-related complications than those treated with ACDF. Rates of transfusion, dysphagia, and hematoma/seroma formation were significantly increased with ACCF compared with ACDF within the elderly population. The elderly are significantly less likely to have a revision surgery after surgical treatment for CSM. Patients treated with ACCF are more likely to need a revision than those treated with ACDF. 3.


Cannabidiol Is a Novel Modulator of Bacterial Membrane Vesicles.
Membrane vesicles (MVs) released from bacteria participate in cell communication and host-pathogen interactions. Roles for MVs in antibiotic resistance are gaining increased attention and in this study we investigated if known anti-bacterial effects of cannabidiol (CBD), a phytocannabinoid from Cannabis sativa, could be in part attributed to effects on bacterial MV profile and MV release. We found that CBD is a strong inhibitor of MV release from Gram-negative bacteria (E. coli VCS257), while inhibitory effect on MV release from Gram-positive bacteria (S. aureus subsp. aureus Rosenbach) was negligible. When used in combination with selected antibiotics, CBD significantly increased the bactericidal action of several antibiotics in the Gram-negative bacteria. In addition, CBD increased antibiotic effects of kanamycin in the Gram-positive bacteria, without affecting MV release. CBD furthermore changed protein profiles of MVs released from E. coli after 1 h CBD treatment. Our findings indicate that CBD may pose as a putative adjuvant agent for tailored co-application with selected antibiotics, depending on bacterial species, to increase antibiotic activity, including via MV inhibition, and help reduce antibiotic resistance.


Chlorfenapyr: a pyrrole insecticide for the control of pyrethroid or DDT resistant Anopheles gambiae (Diptera: Culicidae) mosquitoes.
Owing to the development and spread of pyrethroid resistance in Anopheles gambiae in Africa there is an urgent need to develop alternative insecticides to supplement the pyrethroids. Chlorfenapyr is a pyrrole insecticide first commercialized for the control of agricultural pests and termites. Performance against An. gambiae bearing kdr (pyrethroid and DDT resistance) or Ace-1(R) insensitive acetylcholinesterase (organophosphate and carbamate resistance) mechanisms was studied using a variety of adult bioassay tests including a simulated-experimental hut system (tunnel tests) that allows uninhibited mosquito behaviour/insecticide interactions. Strains resistant to pyrethroids and organophosphates showed no cross resistance to chlorfenapyr. In cone bioassays on treated netting the mortality of adult mosquitoes showed an unexpected curvilinear response, with highest mortality occurring at intermediate dosages. Adults expressed irritability to chlorfenapyr at higher dosages, which might explain the dosage-mortality trend. Toxic activity of chlorfenapyr was slow compared to conventional neurotoxic insecticides and additional mortality occurred between 24h and 72 h. In tunnel tests, the dosage-mortality trend showed a more typical sigmoid response and most mortality occurred during the first 24h. Mosquito penetration through the holed, treated netting showed only limited inhibition and blood-feeding was not inhibited. Mortality rates in the kdr strain exposed to chlorfenapyr treated netting in tunnel tests were much higher than with permethrin treated netting over the same 100-500 mg/m(2) dosage range. Chlorfenapyr has potential for malaria control in treated-net or residual spraying applications in areas where mosquitoes are pyrethroid resistant. For treated-net applications chlorfenapyr might be combined with pyrethroid as a mixture to provide personal protection as well as to give control of resistant mosquitoes.


BMP4 inhibits PDGF-induced proliferation and collagen synthesis via PKA-mediated inhibition of calpain-2 in pulmonary artery smooth muscle cells.
In the present study, we investigated the effect of bone morphogenetic protein 4 (BMP4) on PDGF-induced cell proliferation and collagen synthesis in pulmonary artery smooth muscle cells (PASMCs). Normal human PASMCs were incubated with and without PDGF-BB in the absence and presence of BMP4 for 0.5 to 24 h. The protein levels of collagen-I, p-Smad2/3, p-Smad1/5, and intracellular active TGF-β1, calpain activity, and cell proliferation were then measured. The results showed that BMP4 induced an increase in p-Smad1/5 but had no effect on the protein levels of collagen-I, p-Smad2/3, and intracellular active TGF-β1 and calpain activity in control PASMCs. Nevertheless, BMP4 attenuated increases in cell proliferation and protein levels of collagen-I, p-Smad2/3, and intracellular active TGF-β1 and calpain activity in PASMCs exposed to PDGF-BB. Moreover, BMP4 increased PKA activity and inhibition of PKA prevented the inhibitory effects of BMP4 on PDGF-BB-induced calpain activation in normal PASMCs. The PKA activator forskolin recapitulated the suppressive effect of BMP4 on PDGF-induced calpain activation. Furthermore, BMP4 prevented a PDGF-induced decrease in calpain-2 phosphorylation at serine-369 in normal PASMCs. Finally, BMP4 did not attenuate PDGF-induced increases in cell proliferation, collagen-I protein levels, and calpain activation and did not induce PKA activation and did not prevent a PDGF-induced decrease in calpain-2 phosphorylation at serine-369 in PASMCs from idiopathic pulmonary arterial hypertension (PAH) patients. These data demonstrate that BMP4 inhibits PDGF-induced cell proliferation and collagen synthesis via PKA-mediated inhibition of calpain-2 in normal PASMCs. The inhibitory effects of BMP4 on PDGF-induced cell proliferation, collagen synthesis, and calpain-2 activation are impaired in PASMCs from PAH patients, which may contribute to pulmonary vascular remodeling in PAH.


Value-based Total Hip and Knee Arthroplasty: A Framework for Understanding the Literature.
Since passage of the Patient Protection and Affordable Care Act of 2010, the current decade has witnessed an explosion of the value-based total hip and knee arthroplasty literature. Total hip arthroplasty and total knee arthroplasty are the most common inpatient surgeries for Medicare beneficiaries, and thus, it is no surprise that total joint arthroplasty is currently a prime target of efforts toward cost reduction and quality improvement. The purpose of this review was to provide a framework for understanding the rapidly growing quality and cost literature. Research efforts toward quality improvement are likely to be effective when they address the structure, process, and most importantly outcomes of total joint arthroplasty. Similarly, cost savings should be evaluated with an understanding of existing accounting methods, relationships to the entire cycle of osteoarthritis care, and the direct effect on the quality of care provided.


Pre- and post-bronchodilator airway obstruction are associated with similar clinical characteristics but different prognosis - report from a population-based study.
According to guidelines, the diagnosis of COPD should be confirmed by post-bronchodilator (post-BD) airway obstruction on spirometry; however, in clinical practice, this is not always performed. The aim of this population-based study was to compare clinical characteristics and prognosis, assessed as mortality, between subjects with airway obstruction divided into pre- but not post-BD obstruction, post-BD airway obstruction (COPD), and subjects without airway obstruction. In 2002-2004, four adult population-based cohorts were reexamined with spirometry and structured interview. Subjects with airway obstruction, with a ratio of forced expiratory volume in 1 s to (forced) vital capacity <0.70 (n=993), were identified together with sex- and age-matched referents (n=993). These subjects were further divided into subjects with pre- but not post-BD airway obstruction (pre- not post-BD obstruction) and subjects with post-BD airway obstruction (COPD). Mortality data were collected until December 31, 2014. Out of 993 subjects with airway obstruction, 736 (74%) had COPD and 257 (26%) pre- not post-BD obstruction. Any respiratory symptoms, allergic rhinitis, asthma, exacerbations, and comorbidities were equally common among subjects with COPD and pre- not post-BD obstruction, but less common among nonobstructive subjects. Mortality was highest among subjects with COPD and higher in men than in women. In both sexes, COPD, but not pre- not post-BD obstruction, was associated with an increased risk for death compared to those without airway obstruction. When COPD was divided into Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages, GOLD 2 and 3-4 had an increased risk for death when compared to the nonobstructive group, also when adjusted for common confounders and comorbidities such as heart disease, diabetes, and anxiety/depression. Even though subjects with COPD and pre- not post-BD obstruction had fairly similar presentation of clinical characteristics, only those with COPD, specifically GOLD stage ≥2, had increased risk for death when compared with nonobstructive subjects.


Different cognitive processes underlie human mate choices and mate preferences.
Based on undergraduates' self-reports of mate preferences for various traits and self-perceptions of their own levels on those traits, Buston and Emlen [Buston PM, Emlen ST (2003) Proc Natl Acad Sci USA 100:8805-8810] concluded that modern human mate choices do not reflect predictions of tradeoffs from evolutionary theory but instead follow a "likes-attract" pattern, where people choose mates who match their self-perceptions. However, reported preferences need not correspond to actual mate choices, which are more relevant from an evolutionary perspective. In a study of 46 adults participating in a speed-dating event, we were largely able to replicate Buston and Emlen's self-report results in a pre-event questionnaire, but we found that the stated preferences did not predict actual choices made during the speed-dates. Instead, men chose women based on their physical attractiveness, whereas women, who were generally much more discriminating than men, chose men whose overall desirability as a mate matched the women's self-perceived physical attractiveness. Unlike the cognitive processes that Buston and Emlen inferred from self-reports, this pattern of results from actual mate choices is very much in line with the evolutionary predictions of parental investment theory.


In silico designing and molecular docking of a potent analog against Staphylococcus aureus porphobilinogen synthase.
The emergence of multidrug-resistant strains of Staphylococcus aureus, there is an urgent need for the development of new antimicrobials which are narrow and pathogen specific. In this context, the present study is aimed to have a control on the staphylococcal infections by targeting the unique and essential enzyme; porphobilinogen synthase (PBGS) catalyzes the condensation of two molecules of δ-aminolevulinic acid, an essential step in the tetrapyrrole biosynthesis. Hence developing therapeutics targeting PBGS will be the promising choice to control and manage the staphylococcal infections. 4,5-dioxovalerate (DV) is known to inhibit PBGS. In view of this, in this study, novel dioxovalerate derivatives (DVDs) molecules were designed so as to inhibit PBGS, a potential target of S. aureus and their inhibitory activity was predicted using molecular docking studies by molecular operating environment. The 3D model of PBGS was constructed using Chlorobium vibrioform (Protein Data Bank 1W1Z) as a template by homology modeling method. The built structure was close to the crystal structure with Z score - 8.97. Molecular docking of DVDs into the S. aureus PBGS active site revealed that they are showing strong interaction forming H-bonds with the active sites of K248 and R217. The ligand-receptor complex of DVD13 showed a best docking score of - 14.4555 kcal/mol among DV and all its analogs while the substrate showed docking score of - 13.0392 kcal/mol showing interactions with S199, K217 indicating that DVD13 can influence structural variations on the enzyme and thereby inhibiting the enzyme. The substrate analog DVD13 is showing significant interactions with active site of PBGS and it may be used as a potent inhibitor to control S. aureus infections.


Halogenated hydrocarbons in New Orleans drinking water and blood plasma.
Volatile organics from New Orleans drinking water and pooled plasma were collected on a solid phenyl ether polymer and analyzed by gas chromatographic and mass spectrometric techniques. Thirteen halogenated hydrocarbons were identified in the drinking water. Five halogenated compounds were found in the plasma. Tetrachloroethylene and carbon tetrachloride were found in both the plasma and the drinking water. Considerable variation in the relative concentrations of the halogenated hydrocarbons was noted from day to day in the drinking water.


A chondroitin sulfate proteoglycan may influence the direction of retinal ganglion cell outgrowth.
In the developing retina, retinal ganglion cell (RGC) axons elongate toward the optic fissure, even though no obvious directional restrictions exist. Previous studies indicate that axon-matrix interactions are important for retinal ganglion cell axon elongation, but the factors that direct elongation are unknown. Chondroitin sulfate proteoglycan (CS-PG), a component of the extracellular matrix, repels elongating dorsal root ganglion (DRG) axons in vitro and is present in vivo in the roof plate of the spinal cord, a structure that acts as a barrier to DRG axons during development. In this study, we examined whether CS-PG may regulate the pattern of retinal ganglion cell outgrowth in the developing retina. Immunocytochemical analysis showed that CS-PG was present in the innermost layers of the developing rat retina. The expression of CS-PG moved peripherally with retinal development, always remaining at the outer edge of the front of the developing axons. CS-PG was no longer detectable with immunocytochemical techniques when RGC axon elongation in the retina is complete. Results of studies in vitro showed that CS-PG, isolated from bovine nasal cartilage and chick limb, was inhibitory to elongating RGC axons and that RGC growth cones were more sensitive to CS-PG than were DRG neurites tested at the same concentrations of CS-PG. The behavior of retinal growth cones as they encounter CS-PG was characterized using time-lapse video microscopy. Filopodia of the RGC growth cones extended to and sampled the CS-PG repeatedly. With time, the growth cones turned to avoid outgrowth on the CS-PG and grew only on laminin. While numerous studies have shown the presence of positive factors within the retina that may guide developing RGC axons, this is the first demonstration of an inhibitory or repelling molecule in the retina that may regulate axon elongation. Taken together, these data suggest that the direction of RGC outgrowth in the retina may be regulated by the proper ratio of growth-promoting molecules, such as laminin, to growth-inhibiting molecules, like CS-PG, present in the correct pattern and concentrations along the retinal ganglion cell pathway.


Is a woman's preference for chest hair in men influenced by parasite threat?
Humans (Homo sapiens) are unique primates due to a lack of a thermally insulating fur covering, typical of all other primates. Our primary goal was to examine the "ectoparasite avoidance mediated by mate choice hypothesis" suggesting that women prefer men lacking chest hair in order to avoid ectoparasite loads. We predicted that women living in areas with high prevalence of pathogens (n = 161) would be less likely to show a preference for a male with chest hair in comparison with women living in areas with low pathogen prevalence (n = 183). We found that overall preference for man chest hair was low, but there were no significant associations between perceived vulnerability to diseases or disgust sensitivity and preference of men who have had experimentally removed chest hair. Women who lived in an environment with a high parasite prevalence rate (Turkey) showed a similar preference for man chest hair as did women from an environment with low parasite prevalence (Slovakia). The participants biological fathers' chest hair had no significant effect on their preference for men with chest hair. Women living in a high-parasite-prevalence environment reported a higher disgust score in the sexual domain and more recent experiences with illnesses, suggesting that parasites influence sensitivity to sexual disgust. These results provide no support for the ectoparasite avoidance hypothesis mediated by mate choice and suggest that shaved men bodies are preferred more by women.


Androgen receptor CAGn repeat length influences phenotype of 47,XXY (Klinefelter) syndrome.
Klinefelter syndrome (KS; 47,XXY karyotype and variants) is characterized by tall stature and testicular failure, with marked variation in severity of the phenotype. Previous studies have proposed that genetic factors including mosaicism, parental origin of the supernumerary X-chromosome, skewed X inactivation, and androgen receptor (AR) polyglutamine repeat length may contribute to phenotypic variability in KS. The objective of this study was to investigate the roles of these genetic factors in the variability of the KS phenotype. This was a cross-sectional study. The study was performed at a pediatric endocrinology referral clinic. Thirty-five KS boys and men, aged 0.1-39 yr, were studied. There were no interventions. Auxological measurements, biological indices of testicular function, and clinical assessment of muscle tone were the main outcome measures. Genetic studies included karyotyping to detect mosaicism, genotyping of microsatellite markers to determine parental origin of the supernumerary X-chromosome, and genotyping and methylation studies to measure AR polyglutamine (AR CAGn) repeat length and X inactivation ratio. The only genetic factor that significantly influenced the KS phenotype was the AR CAGn repeat length, which was inversely correlated with penile length, a biological indicator of early androgen action. Mosaicism, imprinting, and skewed X inactivation did not account for the variability of the KS phenotype. Normal genetic variation in the AR coding sequence may be clinically significant in the setting of early testicular failure and subnormal circulating testosterone levels, as occur in KS.


Lipoprotein lipase expression in undifferentiated hepatoma cells is regulated by progesterone and protein kinase A.
Recently, it was shown that lipoprotein lipase (LPL) was produced in neonatal but not in adult rat liver. In an attempt to further define the mechanism involved in liver LPL expression, we identified a neonatal mouse hepatoma cell line, BWTG3, capable of producing LPL. The regulation of LPL expression by various extracellular stimuli was investigated in this cell line. Progesterone caused a rise in LPL production by BWTG3 cells. Other hormones tested, such as insulin, glucagon, adrenalin, testosterone, and thyroid hormone, had no effect on LPL production. The effects of progesterone on LPL production showed slow kinetics reaching a maximum 24 h after addition. Cotransfection of a progesterone receptor expression vector with a 5'-LPL-CAT reporter construct resulted in an induction of CAT activity, suggesting that the increase in LPL accumulation after progesterone was linked to transcriptional induction of the LPL gene. Stimuli causing an elevation of protein kinase A activity in the cells also increased LPL production. Three agents capable of elevating intracellular cAMP levels, i.e., forskolin, dBcAMP, and choleratoxin, caused an elevation of LPL production. The increase in LPL activity caused by forskolin and choleratoxin was paralleled by an elevation of LPL mRNA levels, while dBcAMP only induced a small elevation of LPL mRNA levels. The increase in LPL production was shown to be linked to the stimulation of the PKA signal transduction pathway and was apparently transmitted via the transcription factor CREB. No effect of the stimulation of protein kinase C or calcium/calmodulin-dependent kinase on LPL production was detected.(ABSTRACT TRUNCATED AT 250 WORDS)


A Compact Whole-Eye Perfusion System to Evaluate Pharmacologic Responses of Outflow Facility.
To discover novel therapies that lower IOP by increasing aqueous humor outflow facility, ex vivo ocular perfusion systems provide a valuable tool. However, currently available designs are limited by their throughput. Here we report the development of a compact, scalable perfusion system with improved throughput and its validation using bovine and porcine eyes. At a fixed IOP of 6 mm Hg, flow rate was measured by flow sensors. We validated the system by measuring the outflow responses to Y-39983 (a Rho kinase inhibitor), endothelin-1 (ET-1), ambrisentan (an antagonist for endothelin receptor A [ETA]), sphigosine-1-phosphate (S1P), JTE-013 (antagonist for S1P receptor 2 [S1P2]), S-nitroso-N-acetylpenicillamine (SNAP, a nitric oxide [NO] donor), and 3-Morpholino-sydnonimine (SIN-1, another NO donor). The instrument design enabled simultaneous measurements of 20 eyes with a footprint of 1 m2. Relative to vehicle control, Y-39983 increased outflow by up to 31% in calf eyes. On the contrary, ET-1 decreased outflow by up to 79%, a response that could be blocked by pretreatment with ambrisentan, indicating a role for ETA receptors. Interestingly, the effect of ET-1 was also inhibited by up to 70% to 80% by pretreatment with NO donors, SNAP and SIN-1. In addition to testing in calf eyes, similar effects of ET-1 and ambrisentan were observed in adult bovine and porcine eyes. The compact eye perfusion platform provides an opportunity to efficiently identify compounds that influence outflow facility and may lead to the discovery of new glaucoma therapies.


Small heat shock protein as a multifunctional scaffold: integrated tumor targeting and caspase imaging within a single cage.
Protein cages have the potential to serve as biomaterials for the targeted therapeutic and imaging systems. As an effort to exploit small heat shock protein (Hsp) cages as multifunctional biomaterials, we demonstrate that chemically and genetically modified Hsp cages permeate the cells via cancer cell binding and subsequent endocytic internalization and can image caspase activity in the live cells. Moreover, we report here that these functional Hsp cages can be specifically accumulated to tumor tissues of tumor-bearing mice when administered intravenously through the lateral tail vein. These tumor-targeting properties could be explained by the prolonged in vivo circulation and enhanced permeability and retention (EPR) effect as well as the ligand-mediated binding to cancer cells. Furthermore, when combined with the caspase sensing ability, our Hsp cage allows us to monitor the therapeutic evaluation after anticancer drug treatment by imaging the caspase activity within tumors. Therefore, we demonstrate that the Hsp cages have multifunctional scaffolds amenable to genetic and chemical modifications without loss of the cagelike architecture and can be exploited as biomedical materials including drug or imaging agent carriers.


Patterns of lesion development in multiple sclerosis: longitudinal observations with T1-weighted spin-echo and magnetization transfer MR.
We evaluated the appearance of enhancing multiple sclerosis (MS) lesions on unenhanced T1-weighted MR images and the natural course of enhancing MS lesions on serial unenhanced T1-weighted and magnetization transfer (MT) MR images. One hundred twenty-six enhancing lesions were followed up monthly for 6 to 12 months to determine their signal intensity on unenhanced T1-weighted and MT MR images. At the time of initial enhancement, the size of the lesion and the contrast ratio of enhancement were calculated for each enhancing lesion. During follow-up, the contrast ratio on the corresponding unenhanced T1-weighted image was measured, and an MT ratio (MTR) was calculated. Twenty-five enhancing lesions (20%) appeared isointense and 101 lesions (80%) appeared hypointense relative to normal-appearing white matter on unenhanced T1-weighted images. During 6 months of follow-up, four MR patterns of active lesions were detected: initially isointense lesions remained isointense (15%); initially isointense lesions became hypointense (5%, most of which reenhanced); initially hypointense lesions became isointense (44%); and initially hypointense lesions remained hypointense (36%). MTR was significantly lower for hypointense lesions as compared with isointense lesions at the time of initial enhancement. For lesions that changed from hypointense to isointense, MTR increased significantly during 6 months of follow-up. Multiple regression analysis showed that strongly decreased MTR at the time of initial enhancement and enhancement duration of more than one scan were predictive of a hypointense appearance on unenhanced T1-weighted images at 6 months' follow-up. Ring enhancement was found to be the only (weak) predictor of persistently hypointense signal intensity. Most enhancing lesions appear slightly to significantly hypointense on unenhanced T1-weighted images. Although most hypointensities are reversible, only those lesions that fail to recover on unenhanced T1-weighted and MT images may have considerable irreversible structural changes.


Comparative value of clinical, cytological, and histopathological features in feline mammary gland tumors; an experimental model for the study of human breast cancer.
The diagnosis of breast lesions is usually confirmed by fine-needle aspiration cytology (FNAC) or histological biopsy. Although there is increasing literature regarding the advantages and limitations of both modalities, there is no literature regarding the accuracy of these modalities for diagnosing breast lesions in high-risk patients, who usually have lesions detected by screening. Moreover, few studies have been published regarding the cytopathology of mammary tumors in cats despite widespread use of the animal model for breast cancer formation and inhibition. The objective of the present study was to evaluate the diagnostic interest of cytological and histopathological analysis in feline mammary tumours (FMTs), in order to evaluate its possible value as an animal model. The study was performed in 3 female cats submitted to surgical resections of mammary tumours. The mammary tumours were excised by simple mastectomy or regional mastectomy, with or without the superficial inguinal lymph nodes. Female cats were of different breeds (1 siamese and 2 persians). Before surgical excision of the tumour, FNA cytology was performed using a 0.4 mm diameter needle attached to a 8 ml syringe held in a standard metal syringe holder. The cytological sample was smeared onto a glass slide and either air-dried for May-Grünwald-stain and masses were surgically removed, the tumours were grossly examined and tissue samples were fixed in 10%-buffered-formalin and embedded in paraffin. Sections 4 μm thick were obtained from each sample and H&E stained. Cytologically, atypical epithelial cells coupled to giant nucleus, chromatin anomalies, mitotic figures, spindle shape cells, anisocytosis with anisokaryosis and hyperchromasia were found. Histologically, these tumors are characterized by pleomorphic and polygonal cell population together with mitotic figures, necrotic foci and various numbers inflammatory foci. Also, spindle shaped cells, haemorrhage localized in the different regions, local invasiveness and enlarged nuclei were observed. The samples included 3 tumors of mammary glands mammary tumors were complex carcinomas (n = 2) and adenocarcinoma (n = 1). The histological grades of the 3 cases were as follows: grade II, (1/3); grade III, (2/3) with high mitotic index. The preferential localization of mammary neoplasms was in the inguinal lobe (1/3 case) and abdominal lobes (2/3 cases). Furthermore, 1case of the inguinal mass affected the left caudo-inguinal lobe and 2cases right cranio and caudo abdominal lobes. The study concluded that cytology could be used as a quick, rapid, field diagnostic technique in combination with histopathology for the diagnosis of feline mammary tumors (FMTs). Our findings in feline MTs indicate that FMTs could be useful as an animal model of human breast cancer. Moreover, because of the similarity of the cytohistopathological findings in the human and feline mammary gland tumours, it is possible to use the same cytopathological criteria applied in human pathology for the diagnosis of feline mammary gland tumours. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2047361423103295.


Relationships between post operative pain management and short term functional mobility in total knee arthroplasty patients with a femoral nerve catheter: a preliminary study.
Effective pain management following total knee arthroplasty (TKA) is fundamental in achieving positive rehabilitation outcomes. The purpose of our study was to investigate post operative pain management in relation to short term functional mobility in an intervention group receiving concomitant use of an IV narcotic PCA and a continuous infusion of local anaesthetic via a femoral nerve catheter (CFNC), compared to a group receiving narcotic PCA alone. This was a preliminary study conducted to establish an appropriate design for a larger investigative study. A prospective design was used to measure the effect of a CFNC on post operative pain management and functional mobility prior to hospital discharge. The amount of fentanyl used, pain and nausea scores, timed up and go (TUG) tests and active range of knee movement (AROM) were used to compare a CFNC and supplemental narcotic patient controlled analgesia (PCA) group (n = 27) with a PCA only group (n = 25). The CFNC group used significantly less fentanyl than the PCA only group (p < .001) but there was no significant difference in TUG times between the two groups. There was however a significantly lower AROM reported for both extension (p < .04) and flexion (p < .006,) in the FNC group. Women had significantly slower TUG times (p < .005,) and there were moderate to strong positive correlations between post operative TUG times and the preoperative TUG time (r(s) = .505 p < .001), the time since oral analgesia (r(s) = .529 p < .014), and pain scores (r(s) = .328, p = .034) In this small preliminary study improved TUG performance at Day 4 post op was not influenced by the use of a CFNC but was positively correlated with male gender, preoperative performance, time elapsed since last oral analgesia and pain score. However AROM was decreased in the CFNC group suggesting further research on the relationship between CFNCs, local anaesthetic concentration and quadriceps strength should be incorporated in the follow up study's design.


Ypt32 recruits the Sec4p guanine nucleotide exchange factor, Sec2p, to secretory vesicles; evidence for a Rab cascade in yeast.
SEC2 is an essential gene required for polarized growth of the yeast Saccharomyces cerevisiae. It encodes a protein of 759 amino acids that functions as a guanine nucleotide exchange factor for the small GTPase Sec4p, a regulator of Golgi to plasma membrane transport. Activation of Sec4p by Sec2p is needed for polarized transport of vesicles to exocytic sites. Temperature-sensitive (ts) mutations in sec2 and sec4 result in a tight block in secretion and the accumulation of secretory vesicles randomly distributed in the cell. The proper localization of Sec2p to secretory vesicles is essential for its function and is largely independent of Sec4p. Although the ts mutation sec2-78 does not affect nucleotide exchange activity, the protein is mislocalized. Here we present evidence that Ypt31/32p, members of Rab family of GTPases, regulate Sec2p function. First, YPT31/YPT32 suppress the sec2-78 mutation. Second, overexpression of Ypt31/32p restores localization of Sec2-78p. Third, Ypt32p and Sec2p interact biochemically, but Sec2p has no exchange activity on Ypt32p. We propose that Ypt32p and Sec4p act as part of a signaling cascade in which Ypt32p recruits Sec2p to secretory vesicles; once on the vesicle, Sec2p activates Sec4p, enabling the polarized transport of vesicles to the plasma membrane.


Administration of palmitoylethanolamide (PEA) protects the neurovascular unit and reduces secondary injury after traumatic brain injury in mice.
Traumatic brain injury (TBI) is a major cause of preventable death and morbidity in young adults. This complex condition is characterized by significant blood brain barrier leakage that stems from cerebral ischemia, inflammation, and redox imbalances in the traumatic penumbra of the injured brain. Recovery of function after TBI is partly through neuronal plasticity. In order to test whether treatments that enhance plasticity might improve functional recovery, a controlled cortical impact (CCI) in adult mice, as a model of TBI, in which a controlled cortical impactor produced full thickness lesions of the forelimb region of the sensorimotor cortex, was performed. Once trauma has occurred, combating these exacerbations is the keystone of an effective TBI therapy. The endogenous fatty acid palmitoylethanolamide (PEA) is one of the members of N-acyl-ethanolamines family that maintain not only redox balance but also inhibit the mechanisms of secondary injury. Therefore, we tested whether PEA shows efficacy in a mice model of experimental TBI. PEA treatment is able to reduced edema and brain infractions as evidenced by decreased 2,3,5-triphenyltetrazolium chloride staining across brain sections. PEA-mediated improvements in tissues histology shown by reduction of lesion size and improvement in apoptosis level further support the efficacy of PEA therapy. The PEA treatment blocked infiltration of astrocytes and restored CCI-mediated reduced expression of PAR, nitrotyrosine, iNOS, chymase, tryptase, CD11b and GFAP. PEA inhibited the TBI-mediated decrease in the expression of pJNK and NF-κB. PEA-treated injured animals improved neurobehavioral functions as evaluated by behavioral tests.


Development and validation of the Day in the Life Questionnaire (DILQ) as a measure of fruit and vegetable questionnaire for 7-9 year olds.
The Day in the Life Questionnaire (DILQ) was developed as a supervised classroom exercise to measure children's consumption of fruit and vegetables. The DILQ uses words and pictures to encourage the child to recall and describe a range of activities from the previous day, including their entire food intake. This study tested the validity and reliability of the DILQ for children aged 7-9 years (n = 255) in four English schools. Reliability, validity and sensitivity to change were assessed through repeated rounds of data collection. Comparisons were made of observations during school breaks and classroom completion of the DILQ. Children enjoyed completing the DILQ and teachers thought it appropriate for the age group. The questionnaire performed either well or acceptably on all validity, reliability and sensitivity tests. The DILQ can be recommended as a method of collecting data for fruit and vegetable consumption from children aged 7-9 in the classroom. The validation study included comparison of schools with and without 'fruit only' breaktime policies, and sensitivity to a brief intervention in which free fruit was distributed at morning break. The results suggest that it would be a sensitive measure for descriptive studies, before and after studies and controlled trials.


2-MeOE2bisMATE and 2-EtE2bisMATE induce cell cycle arrest and apoptosis in breast cancer xenografts as shown by a novel ex vivo technique.
Breast cancer is the leading cause of cancer deaths among women worldwide. The theory of targeting both cancer cells directly and their blood supply has significant therapeutic potential. However, to date, there are few clinically successful single agents that meet these criteria. 2-Methoxyestradiol-3,17-O,O-bis-sulfamate (2-MeOE2bisMATE) and 2-ethylestradiol-3,17-O,O-bis-sulfamate (2-EtE2bisMATE) are potent inhibitors of proliferation in a range of cancer cells. The work presented here demonstrates the potent in vitro and in vivo effects of these compounds. They cause apoptosis via the intrinsic mitochondrial pathway in both MDA-MB-231 breast cancer cells and endothelial cells. Furthermore, they are potent anti-angiogenic inhibitors in vivo, as shown by their ability to reduce endothelial staining in MDA-MB-231 xenograft tumors. We have developed a novel, flow cytometry based, ex vivo method which shows in cells recovered from MDA-MB-231 tumors treated with 2-MeOE2bisMATE and 2-EtE2bisMATE an increase in intra-tumoral G(2)-M arrest and apoptosis. The degree of apoptosis inversely correlates to tumor volume. Further in vivo studies reveal that both 2-MeOE2bisMATE and 2-EtE2bisMATE are orally bioavailable and extremely efficacious when compared to clinically tested drugs. As these compounds are anti-proliferative against breast cancer and endothelial cells they have the potential to be potent, dual acting clinical drugs of the future.


Characterization of cellular and humoral immune responses after IBV infection in chicken lines differing in MBL serum concentration.
Chickens from two inbred lines selected for high (L10H) or low (L10L) mannose-binding lectin (MBL) serum concentrations were infected with infectious bronchitis virus (IBV), and innate as well as adaptive immunological parameters were measured throughout the experimental period. Chickens with high MBL serum concentrations were found to have less viral load in the trachea than chickens with low MBL serum concentrations indicating that these chickens were less severely affected by the infection. This study is the first to show that MBL expression is present in the lungs of healthy chickens and that the expression is upregulated at days 3 postinfection (p.i.) in L10H chickens. Furthermore, in the liver of infected chickens, the MBL expression was upregulated at day 7 p.i., despite the fact that the MBL serum concentrations were decreased below baseline at that time point. The number of TCRγδ+CD8α+ cells in the blood of noninfected chickens increased from week 0 to 3 p.i. However, the number of cells was higher in L10H chickens than in L10L chickens throughout the experiment. No increase was observed in the number of TCRγδ+CD8α+ cells in the blood of the infected L10H and L10L chickens. The numbers of B cells at week 3 p.i. were higher for noninfected L10L chickens than for the other chickens. No differences were observed between the infected and noninfected L10H chickens or between the infected L10H and L10L chickens. Furthermore, at week 3 p.i., the number of monocytes was higher in infected and noninfected L10H chickens than in the infected and noninfected L10L chickens. Thus, these results indicate that MBL is produced locally and may be involved in the regulation of the cellular immune response after an IBV infection. However, MBL did not appear to influence the humoral immune response after IBV infection in this study.


Essential role for centromeric factors following p53 loss and oncogenic transformation.
In mammals, centromere definition involves the histone variant CENP-A (centromere protein A), deposited by its chaperone, HJURP (Holliday junction recognition protein). Alterations in this process impair chromosome segregation and genome stability, which are also compromised by p53 inactivation in cancer. Here we found that CENP-A and HJURP are transcriptionally up-regulated in p53-null human tumors. Using an established mouse embryonic fibroblast (MEF) model combining p53 inactivation with E1A or HRas-V12 oncogene expression, we reproduced a similar up-regulation of HJURP and CENP-A. We delineate functional CDE/CHR motifs within the Hjurp and Cenpa promoters and demonstrate their roles in p53-mediated repression. To assess the importance of HJURP up-regulation in transformed murine and human cells, we used a CRISPR/Cas9 approach. Remarkably, depletion of HJURP leads to distinct outcomes depending on their p53 status. Functional p53 elicits a cell cycle arrest response, whereas, in p53-null transformed cells, the absence of arrest enables the loss of HJURP to induce severe aneuploidy and, ultimately, apoptotic cell death. We thus tested the impact of HJURP depletion in pre-established allograft tumors in mice and revealed a major block of tumor progression in vivo. We discuss a model in which an "epigenetic addiction" to the HJURP chaperone represents an Achilles' heel in p53-deficient transformed cells.


Impact of nanoparticles on the CO2-brine interfacial tension at high pressure and temperature.
Nanofluid flooding has been identified as a promising method for enhanced oil recovery (EOR) and improved Carbon geo-sequestration (CGS). However, it is unclear how nanoparticles (NPs) influence the CO2-brine interfacial tension (γ), which is a key parameter in pore-to reservoirs-scale fluid dynamics, and consequently project success. The effects of pressure, temperature, salinity, and NPs concentration on CO2-silica (hydrophilic or hydrophobic) nanofluid γ was thus systematically investigated to understand the influence of nanofluid flooding on CO2 geo-storage. Pendant drop method was used to measure CO2/nanofluid γ at carbon storage conditions using high pressure-high temperature optical cell. CO2/nanofluid γ was increased with temperature and decreased with increased pressure which is consistent with CO2/water γ. The hydrophilicity of NPs was the major factor; hydrophobic silica NPs significantly reduced γ at all investigated pressures and temperatures while hydrophilic NPs showed only minor influence on γ. Further, increased salinity which increased γ can also eliminate the influence of NPs on CO2/nanofluid γ. Hence, CO2/brine γ has low, but, reasonable values (higher than 20 mN/m) at carbon storage conditions even with the presence of hydrophilic NPs, therefore, CO2 storage can be considered in oil reservoirs after flooding with hydrophilic nanofluid. The findings of this study provide new insights into nanofluids applications for enhanced oil recovery and carbon geosequestration projects.


Herpes zoster ophthalmicus: declining age at presentation.
To investigate changes in the age of occurrence of herpes zoster ophthalmicus (HZO) in patients presenting to the Massachusetts Eye and Ear Infirmary (MEEI) from 2007 through 2013. Retrospective chart review. Academic tertiary referral centre for ophthalmic conditions. 913 patients with acute HZO. A total of 1283 potential cases were identified by searching the MEEI electronic medical record for patient charts with International Classification of Diseases 9 codes for herpes zoster, shingles and varicella from 2007 through 2013. The cases were reviewed to confirm diagnosis of acute HZO, requiring documentation of a skin rash or pain in the V1 distribution, resulting in inclusion of 913 cases. Number of HZO cases each year, mean age of HZO cases each year, number of HZO cases with an immunodeficiency state. The number of patients with HZO presenting to MEEI increased from 71 cases in 2007 to 195 cases in 2013. The mean age of patients with acute HZO reduced significantly from 61.2 years in 2007 to 55.8 years in 2013 (p=0.0119). The number of patients with acute HZO in the setting of an immunodeficiency state did not change significantly over the study period. Ever since the introduction of varicella vaccination in children, there has been debate regarding its effect on zoster epidemiology, particularly regarding the potential to reduce population exposure and limit repeated immunological boosts against varicella zoster virus in adults. Patients presenting to MEEI with HZO were younger on average in 2013 than in 2007. Although a population-based study is necessary to test the hypothesis, our study suggests that varicella vaccination of children remains a possible explanation for the increased number of cases and reduction in mean age of newly diagnosed patients.


Tromp1, a putative rare outer membrane protein, is anchored by an uncleaved signal sequence to the Treponema pallidum cytoplasmic membrane.
Treponema pallidum rare outer membrane protein 1 (Tromp1) has extensive sequence homology with substrate-binding proteins of ATP-binding cassette transporters. Because such proteins typically are periplasmic or cytoplasmic membrane associated, experiments were conducted to clarify Tromp1's physicochemical properties and cellular location in T. pallidum. Comparison of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis mobilities of (i) native Tromp1 and Tromp1 synthesized by coupled in vitro transcription-translation and (ii) native Tromp1 and recombinant Tromp1 lacking the N-terminal signal sequence revealed that the native protein is not processed. Other studies demonstrated that recombinant Tromp1 lacks three basic porin-like properties: (i) the ability to form aqueous channels in liposomes which permit the influx of small hydrophilic solutes, (ii) an extensive beta-sheet secondary structure, and (iii) amphiphilicity. Subsurface localization of native Tromp1 was demonstrated by immunofluorescence analysis of treponemes encapsulated in gel microdroplets, while opsonization assays failed to detect surface-exposed Tromp1. Incubation of motile treponemes with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)-diazarine, a photoactivatable, lipophilic probe, also did not result in the detection of Tromp1 within the outer membranes of intact treponemes but, instead, resulted in the labeling of a basic 30.5-kDa presumptive outer membrane protein. Finally, analysis of fractionated treponemes revealed that native Tromp1 is associated predominantly with cell cylinders. These findings comprise a body of evidence that Tromp1 actually is anchored by an uncleaved signal sequence to the periplasmic face of the T. pallidum cytoplasmic membrane, where it likely subserves a transport-related function.


Temporal matching among diurnal photosynthetic patterns within the crown of the evergreen sclerophyll Olea europaea L.
Trees are modular organisms that adjust their within-crown morphology and physiology in response to within-crown light gradients. However, whether within-plant variation represents a strategy for optimizing light absorption has not been formally tested. We investigated the arrangement of the photosynthetic surface throughout one day and its effects on the photosynthetic process, at the most exposed and most sheltered crown layers of a wild olive tree (Olea europaea L.). Similar measurements were made for cuttings taken from this individual and grown in a greenhouse at contrasted irradiance-levels (100 and 20% full sunlight). Diurnal variations in light interception, carbon fixation and carbohydrate accumulation in sun leaves were negatively correlated with those in shade leaves under field conditions when light intensity was not limiting. Despite genetic identity, these complementary patterns were not found in plants grown in the greenhouse. The temporal disparity among crown positions derived from specialization of the photosynthetic behaviour at different functional and spatial scales: architectural structure (crown level) and carbon budget (leaf level). Our results suggest that the profitability of producing a new module may not only respond to construction costs or light availability, but also rely on its spatio-temporal integration within the productive processes at the whole-crown level.


The physician-scientists: rare species in Africa.
There is paucity of physician-scientists in Africa, resulting in overt dependence of clinical practice on research findings from advanced "first world" countries. Physician-scientists include individuals with a medical degree alone or combined with other advanced degrees (e.g. MD/MBChB and PhD) with a career path in biomedical/ translational and patient-oriented/evaluative science research. The paucity of clinically trained research scientists in Africa could result in dire consequences as exemplified in the recent Ebola virus epidemic in West Africa, where shortage of skilled clinical scientists, played a major role in disease progression and mortality. Here we contextualise the role of physician-scientist in health care management, highlight factors limiting the training of physician-scientist in Africa and proffer implementable recommendations to address these factors.


Paediatric single mitochondrial DNA deletion disorders: an overlapping spectrum of disease.
Single large-scale mitochondrial DNA (mtDNA) deletions (SLSMDs) are amongst the most frequently diagnosed mtDNA disorders in childhood, yet their natural history remains poorly understood. We report the natural history of a large multicentre cohort of such children. We reviewed case notes from three different UK centres to determine the clinical course of 34 patients (16 female, 18 male) with childhood-onset mitochondrial disease caused by SLSMDs. Kaplan-Meier analysis was used to compare survival of patients presenting with haematological features (Pearson syndrome) and those with nonhaematological presentations. The most frequent initial presentation was with isolated ptosis (16/34, 47%). Eleven (32%) patients presented with transfusion-dependent anaemia soon after birth and were diagnosed with Pearson syndrome, whilst ten were classified as having Kearns-Sayre syndrome, three as having progressive external ophthalmoplegia (PEO) and seven as having PEO-plus. Three patients did not conform to any specific mitochondrial syndrome. The most frequently affected organ during the disease course was the kidney, with documented tubular or glomerular dysfunction in 17 of 20 (85%) cases who had detailed investigations. SLSMDs were present in blood and/or urine cells in all cases tested, indicating that muscle biopsy is not necessary for diagnosis in the paediatric age range. Kaplan-Meier survival analysis revealed significantly worse mortality in patients with Pearson syndrome compared with the rest of the cohort. Mitochondrial disease caused by SLSMDs is clinically heterogeneous, and not all cases conform to a classical mitochondrial syndrome. Multisystem disease is the norm, with anaemia, renal impairment and endocrine disturbance being the most frequent extraneurological features. SLSMDs should be considered in the differential diagnosis of all children presenting with ptosis.


The deoxyribonucleoside phosphotransferase of Trichomonas vaginalis. A potential target for anti-trichomonial chemotherapy.
Trichomonas vaginalis, a human protozoan parasite known to lack the capability of synthesizing purine and pyrimidine nucleotides de novo, was found also incapable of converting its ribonucleotides to deoxyribonucleotides. The only apparent means of providing deoxyribonucleotides for DNA synthesis relies on salvaging exogenous deoxyribonucleosides by a deoxyribonucleoside phosphotransferase activity in the T. vaginalis 10(5) g pelletable fraction. The activity, constituted by at least two isozymes I and II, can be solubilized by Triton X-100, has a pH optimum of 5.0-6.0, and recognizes only thymidine, deoxyadenosine, deoxyguanosine, and deoxycytidine as the phosphate acceptor. TMP, dAMP, dGMP, dCMP, dUMP, FdUMP, and p-nitrophenylphosphate can serve as phosphate donors. Enzyme I has been purified 10-fold by DEAE-Sepharose chromatography and Sephacryl 200 filtration, and is totally freed of the acid phosphatase of T. vaginalis. It has an estimated molecular weight of 200,000 and Km values of 2-3 mM for the four deoxyribonucleosides, which act on each other as competitive inhibitors. It also possesses phosphatase activity capable of hydrolyzing p-nitrophenylphosphate with a Michaelis constant of 0.74 mM. The rates of hydrolysis are enhanced by thymidine, which suggests that the latter may be the preferred phosphate acceptor, and Enzyme I may be, thus, more a transferase than a phosphatase. This enzyme could be a potential target for antitrichomonial chemotherapy.


Comparison of 3 T MRI and CT for the measurement of visceral and subcutaneous adipose tissue in humans.
CT is considered the gold standard imaging modality for measurement of visceral adipose tissue area. However, as CT imaging exposes subjects to ionising radiation, a comparable imaging technique without this exposure is desirable, such as MRI. Therefore, we compared the agreement of measures of visceral adipose tissue and subcutaneous adipose tissue area from single-slice images obtained at the umbilicus using a 3 T MRI scanner with single-slice images obtained via CT scan. 64 images were obtained from 27 subjects who underwent MRI and CT scanning on the same day, after 10-12 hours of fasting. Visceral and subcutaneous adipose tissue depots were manually separated and quantified using a multimodality image-processing software program. We found good agreement between CT and MRI for the measurement of both visceral adipose tissue and subcutaneous adipose tissue. Bland-Altman difference analysis demonstrated a mean bias of -2.9% (as a portion of total abdominal area) for visceral adipose tissue and +0.4% for subcutaneous adipose tissue, as measured by MRI compared with CT. MRI is a safe, accurate and precise imaging modality for measuring both visceral and subcutaneous adipose tissue, making it a favourable alternative to CT for quantification of these adipose depots.


Engineered ACE2 receptor traps potently neutralize SARS-CoV-2.
An essential mechanism for SARS-CoV-1 and -2 infection begins with the viral spike protein binding to the human receptor protein angiotensin-converting enzyme II (ACE2). Here we describe a stepwise engineering approach to generate a set of affinity optimized, enzymatically inactivated ACE2 variants that potently block SARS-CoV-2 infection of cells. These optimized receptor traps tightly bind the receptor binding domain (RBD) of the viral spike protein and prevent entry into host cells. We first computationally designed the ACE2-RBD interface using a two-stage flexible protein backbone design process that improved affinity for the RBD by up to 12-fold. These designed receptor variants were affinity matured an additional 14-fold by random mutagenesis and selection using yeast surface display. The highest affinity variant contained seven amino acid changes and bound to the RBD 170-fold more tightly than wild-type ACE2. With the addition of the natural ACE2 collectrin domain and fusion to a human Fc domain for increased stabilization and avidity, the most optimal ACE2 receptor traps neutralized SARS-CoV-2 pseudotyped lentivirus and authentic SARS-CoV-2 virus with half-maximal inhibitory concentrations (IC50) in the 10-100 ng/ml range. Engineered ACE2 receptor traps offer a promising route to fighting infections by SARS-CoV-2 and other ACE2-utilizing coronaviruses, with the key advantage that viral resistance would also likely impair viral entry. Moreover, such traps can be predesigned for viruses with known entry receptors for faster therapeutic response without the need for neutralizing antibodies isolated or generated from convalescent patients.


Proteomic analysis of the soybean symbiosome identifies new symbiotic proteins.
Legumes form a symbiosis with rhizobia in which the plant provides an energy source to the rhizobia bacteria that it uses to fix atmospheric nitrogen. This nitrogen is provided to the legume plant, allowing it to grow without the addition of nitrogen fertilizer. As part of the symbiosis, the bacteria in the infected cells of a new root organ, the nodule, are surrounded by a plant-derived membrane, the symbiosome membrane, which becomes the interface between the symbionts. Fractions containing the symbiosome membrane (SM) and material from the lumen of the symbiosome (peribacteroid space or PBS) were isolated from soybean root nodules and analyzed using nongel proteomic techniques. Bicarbonate stripping and chloroform-methanol extraction of isolated SM were used to reduce complexity of the samples and enrich for hydrophobic integral membrane proteins. One hundred and ninety-seven proteins were identified as components of the SM, with an additional fifteen proteins identified from peripheral membrane and PBS protein fractions. Proteins involved in a range of cellular processes such as metabolism, protein folding and degradation, membrane trafficking, and solute transport were identified. These included a number of proteins previously localized to the SM, such as aquaglyceroporin nodulin 26, sulfate transporters, remorin, and Rab7 homologs. Among the proteome were a number of putative transporters for compounds such as sulfate, calcium, hydrogen ions, peptide/dicarboxylate, and nitrate, as well as transporters for which the substrate is not easy to predict. Analysis of the promoter activity for six genes encoding putative SM proteins showed nodule specific expression, with five showing expression only in infected cells. Localization of two proteins was confirmed using GFP-fusion experiments. The data have been deposited to the ProteomeXchange with identifier PXD001132. This proteome will provide a rich resource for the study of the legume-rhizobium symbiosis.


Tofacitinib enhances delivery of antibody-based therapeutics to tumor cells through modulation of inflammatory cells.
The routes by which antibody-based therapeutics reach malignant cells are poorly defined. Tofacitinib, an FDA-approved JAK inhibitor, reduced tumor-associated inflammatory cells and allowed increased delivery of antibody-based agents to malignant cells. Alone, tofacitinib exhibited no antitumor activity, but combinations with immunotoxins or an antibody-drug conjugate resulted in increased antitumor responses. Quantification using flow cytometry revealed that antibody-based agents accumulated in malignant cells at higher percentages following tofacitinib treatment. Profiling of tofacitinib-treated tumor-bearing mice indicated that cytokine transcripts and various proteins involved in chemotaxis were reduced compared with vehicle-treated mice. Histological analysis revealed significant changes to the composition of the tumor microenvironment, with reductions in monocytes, macrophages, and neutrophils. Tumor-associated inflammatory cells contributed to non-target uptake of antibody-based therapeutics, with mice treated with tofacitinib showing decreased accumulation of therapeutics in intratumoral inflammatory cells and increased delivery to malignant cells. The present findings serve as a rationale for conducting trials where short-term treatments with tofacitinib could be administered in combination with antibody-based therapies.


Novel Trimodal MALDI Imaging Mass Spectrometry (IMS3) at 10 μm Reveals Spatial Lipid and Peptide Correlates Implicated in Aβ Plaque Pathology in Alzheimer's Disease.
Multimodal chemical imaging using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) can provide comprehensive molecular information in situ within the same tissue sections. This is of relevance for studying different brain pathologies such as Alzheimer's disease (AD), where recent data suggest a critical relevance of colocalizing Aβ peptides and neuronal lipids. We here developed a novel trimodal, high-resolution (10 μm) MALDI imaging MS (IMS) paradigm for negative and positive ion mode lipid analysis and subsequent protein ion imaging on the same tissue section. Matrix sublimation of 1,5-diaminonaphthalene (1,5-DAN) enabled dual polarity lipid MALDI IMS on the same pixel points at high spatial resolutions (10 μm) and with high spectral quality. This was followed by 10 μm resolution protein imaging on the same measurement area, which allowed correlation of lipid signals with protein distribution patterns within distinct cerebellar regions in mouse brain. The demonstrated trimodal imaging strategy (IMS3) was further shown to be an efficient approach for simultaneously probing Aβ plaque-associated lipids and Aβ peptides within the hippocampus of 18 month-old transgenic AD mice (tgArcSwe). Here, IMS3 revealed a strong colocalization of distinct lipid species including ceramides, phosphatidylinositols, sulfatides (Cer 18:0, PI 38:4, ST 24:0) and lysophosphatidylcholines (LPC 16:0, LPC 18:0) with plaque-associated Aβ isoforms (Aβ 1-37, Aβ 1-38, Aβ 1-40). This highlights the potential of IMS3 as an alternative, superior approach to consecutively performed immuno-based Aβ staining strategies. Furthermore, the IMS3 workflow allowed for multimodal in situ MS/MS analysis of both lipids and Aβ peptides. Altogether, the here presented IMS3 approach shows great potential for comprehensive, high-resolution molecular analysis of histological features at cellular length scales with high chemical specificity. It therefore represents a powerful approach for probing the complex molecular pathology of, e.g., neurodegenerative diseases that are characterized by neurotoxic protein aggregation.


Understanding the role of social factors in recovery after hip fractures: A structured scoping review.
Poor recovery among older adults with hip fractures can occur despite successful surgical repair and rehabilitation, suggesting other factors might play a role in recovery, such as social factors. The aim of this scoping review was to provide an overview of the literature on the role of social factors in older adult's recovery after hip fracture. This review followed the York Framework and its modifications and recent reporting guidelines. Two independent researchers searched main medical databases (CINAHL, EMBASE, Medline, PsycINFO and the Cochrane libraries) from inception to June 2017, for studies investigating social factors and recovery post hip fracture. Studies were excluded if they were qualitative, perspective papers or if participants were < 65 years or they were not living in the community. We screened 2,503 unique abstracts in total and 19 studies fulfilled the inclusion criteria. Social factors investigated in the included studies were social support, socioeconomic factors and living arrangement. We classified outcomes in the studies into three subgroups: physical functional recovery, mortality and other outcomes (pain, hospital length of stay and quality of life). We found evidence that social support and socioeconomic factors (e.g. socioeconomic status) were significantly associated with an increase in functional recovery, a decrease in mortality and other outcomes, but conflicting evidence was found for the effect of one's living arrangement. Only two included studies were randomised controlled trials. To conclude, social factors, such as social support and socioeconomic status, affect physical functional recovery and mortality in older adults with hip fractures. However, this is an under researched area that lacks rigorously designed studies and would benefit from more studies with rigorous designs.


Highly divergent mussel lineages in isolated Indonesian marine lakes.
Marine lakes, with populations in landlocked seawater and clearly delineated contours, have the potential to provide a unique model to study early stages of evolution in coastal marine taxa. Here we ask whether populations of the mussel Brachidontes from marine lakes in Berau, East Kalimantan (Indonesia) are isolated from each other and from the coastal mangrove systems. We analyzed sequence data of one mitochondrial marker (Cytochrome Oxidase I (COI)), and two nuclear markers (18S and 28S). In addition, we examined shell shape using a geometric morphometric approach. The Indonesian populations of Brachidontes spp. harbored four deeply diverged lineages (14-75% COI corrected net sequence divergence), two of which correspond to previously recorded lineages from marine lakes in Palau, 1,900 km away. These four lineages also showed significant differences in shell shape and constitute a species complex of at least four undescribed species. Each lake harbored a different lineage despite the fact that the lakes are separated from each other by only 2-6 km, while the two mangrove populations, at 20 km distance from each other, harbored the same lineage and shared haplotypes. Marine lakes thus represent isolated habitats. As each lake contained unique within lineage diversity (0.1-0.2%), we suggest that this may have resulted from in situdivergence due to isolation of founder populations after the formation of the lakes (6,000-12,000 years before present). Combined effects of stochastic processes, local adaptation and increased evolutionary rates could produce high levels of differentiation in small populations such as in marine lake environments. Such short-term isolation at small spatial scales may be an important contributing factor to the high marine biodiversity that is found in the Indo-Australian Archipelago.


Standardization of criteria for adolescent idiopathic scoliosis brace studies: SRS Committee on Bracing and Nonoperative Management.
Literature review. To establish consistent parameters for future adolescent idiopathic scoliosis bracing studies so that valid and reliable comparisons can be made. Current bracing literature lacks consistency for both inclusion criteria and the definitions of brace effectiveness. A total of 32 brace treatment studies and the current bracing in adolescent idiopathic scoliosis proposal were analyzed to: (1) determine inclusion criteria that will best identify those patients most at risk for progression, (2) determine the most appropriate definitions for bracing effectiveness, and (3) identify additional variables that would provide valuable information. Early brace studies lacked clarity in their inclusion criteria. In more recent studies, inclusion criteria have narrowed considerably to include primarily those patients most at risk for curve progression who may benefit from the use of a brace. Brace effectiveness was usually defined by various degrees of curve progression at maturity. Less frequently, it was defined by the resultant curve magnitude at maturity, whether or not surgical intervention was needed, or if there was change to another brace. Optimal inclusion criteria for future adolescent idiopathic scoliosis brace studies consist of: age is 10 years or older when brace is prescribed, Risser 0-2, primary curve angles 25 degrees -40 degrees , no prior treatment, and, if female, either premenarchal or less than 1 year postmenarchal. Assessment of brace effectiveness should include: (1) the percentage of patients who have < or =5 degrees curve progression and the percentage of patients who have > or =6 degrees progression at maturity, (2) the percentage of patients with curves exceeding 45 degrees at maturity and the percentage who have had surgery recommended/undertaken, and (3) 2-year follow-up beyond maturity to determine the percentage of patients who subsequently undergo surgery. All patients, regardless of subjective reports on compliance, should be included in the results (intent to treat). Every study should provide results stratified by curve type and size grouping.


Asymmetric hybridization and gene flow between Joshua trees (Agavaceae: Yucca) reflect differences in pollinator host specificity.
The angiosperms are by far the largest group of terrestrial plants. Their spectacular diversity is often attributed to specialized pollination. Obligate pollination mutualisms where both a plant and its pollinator are dependent upon one another for reproduction are thought to be prone to rapid diversification through co-evolution and pollinator isolation. However, few studies have evaluated the degree to which pollinators actually mediate reproductive isolation in these systems. Here, we examine evidence for hybridization and gene flow between two subspecies of Joshua tree (Yucca brevifolia brevifolia and Yucca brevifolia jaegeriana) pollinated by two sister species of yucca moth. Previous work indicated that the pollinators differ in host specificity, and DNA sequence data suggested asymmetric introgression between the tree subspecies. Through intensive sampling in a zone of sympatry, a large number of morphologically intermediate trees were identified. These included trees with floral characters typical of Y. b. jaegeriana, but vegetative features typical of Y. b. brevifolia. The opposite combination-Y. b. brevifolia flowers with Y. b. jaegeriana vegetative morphology-never occurred. Microsatellite genotyping revealed a high frequency of genetically admixed, hybrid trees. Coalescent-based estimates of migration indicated significant gene flow between the subspecies and that the direction of gene flow matches differences in pollinator host fidelity. The data suggest that pollinator behaviour determines the magnitude and direction of gene flow between the two subspecies, but that specialized pollination alone is not sufficient to maintain species boundaries. Natural selection may be required to maintain phenotypic differences in the face of ongoing gene flow.


Characterization of embryonic stem-like cells derived from HEK293T cells through miR302/367 expression and their potentiality to differentiate into germ-like cells.
Human induced pluripotent stem (iPS) cells have great value for regenerative medicine, but are facing problems of low efficiency. MicroRNAs are a recently discovered class of 19-25 nt small RNAs that negatively target mRNAs. miR302/367 cluster has been demonstrated to reprogram mouse and human somatic cells to iPS cells without exogenous transcription factors, however, the repetition and differentiation potentiality of miR302/367-induced pluripotent stem (mirPS) cells need to be improved. Here, we showed overexpression of miR302/367 cluster reprogrammed human embryonic kidney 293T cells into mirPS cells in serum-free N2B27-based medium. The mirPS cells had similar morphology with embryonic stem cells, and expressed pluripotent markers including Oct4, Sox2, Klf4, and Nanog. In addition, through formation of embryoid bodies, various cells and tissues from three germ layers could be determined. Moreover, we examined the potential of mirPS cells differentiating into germ cells both in vitro and in vivo. Taken together, these data might provide a new source of cells and technique for the investigation of the mechanisms underlying reprogramming and pluripotency.


Perceptive fields of saliency.
Visual saliency plays an important role in early vision. Humans automatically orient to salient information, via covert attentional shifts and overt eye movements. Here, we measured saliency using a novel psychophysical method. The stimulus was a grid of colored oriented lines. Line luminance varied continuously over the image. Using a mouse, participants adjusted line luminance at locations in the image, until all lines appeared homogeneously luminant. Participants tended to increase (or decrease) luminance of lines where perceptually salient information was absent (or present), thus line luminance setting correlates with perceived saliency. Perceptually less salient image regions were more homogenous in color and orientation, consistent with iso-feature suppression. Perceptual fields of contextual modulation are derived, showing increased saliency surrounding color and/or orientation changes, increased saliency for collinear and end-stopping lines, and a nonlinear integration of saliencies across dimensions. It took 3 or more surround items identical to a target to generate a measurable inhibitory effect, beyond which every additional identical item had a monotonic effect. These novel findings allow a revision of current models of visual saliency. In particular, we found evidence of sustained saliency. Moreover, this new method is sensitive within the normal functioning range, unlike most current research methods.


Emotional Support, Depressive Symptoms, and Age-Related Alterations in Male Body Composition: Cross-Sectional Findings from the Men's Health 40+ Study.
More depressive symptoms and low emotional support have been related to worse body composition. Body composition significantly deteriorates in aging men. Therefore, we aimed to examine whether high emotional support and low depressive symptoms are associated with better body composition and a decelerated age-related deterioration of body composition in aging men. A cross-sectional analysis including 269 self-reporting healthy men aged between 40 and 75 years living in the German-speaking part of Switzerland was conducted. Participants completed questionnaires on emotional support and depressive symptoms. The depression screening instrument was used to form a group with low (N = 225) and moderate (N = 44) depressive symptoms. Body mass index (BMI) and waist-to-hip ratio (WHR) were measured, and cell proportion (CP), fat mass (FM), and water balance (WB) were obtained using bioelectrical impedance analysis. Age-related associations emerged for WHR, CP, FM, and WB, but not for BMI. Emotional support was negatively associated with BMI, WHR, and WB, and only trend-wise with CP and FM. Group comparisons revealed that more depressive symptoms were associated with lower levels of CP and higher levels of WB. Both emotional support and depressive symptoms were significant moderators of the association between age and specific measures of body composition such as CP, FM, and WB. However, after correction for multiple testing for moderation analyses only the moderation effects of depressive symptoms on the association between age and WB and CP remained significant. Low depressive symptoms were associated with a better body composition as well as a decelerated decline in body composition and the role of emotional support acting as a buffer against age-related deterioration of body composition merits further investigation.


Kinetic analysis of the triggered exocytosis/endocytosis secretory cycle in cultured bovine adrenal medullary cells.
Cultured bovine adrenal medullary cells are an excellent preparation for quantitative analysis of the secretory exocytosis/endocytosis cycle. In this paper we examine the kinetics of endocytosis after stimulation of secretion. Membrane retrieval was monitored by uptake of the fluid phase marker horseradish peroxidase. Horseradish peroxidase was found to be suitable because it can be washed off completely, assayed quantitatively, and its uptake increases linearly with concentration. If this marker is present during stimulation, the rate of uptake is initially slower than catecholamine secretion but faster at a later time, suggesting that the formation of endocytotic vesicles follows exocytosis. To monitor the time-dependent concentration of secretory vesicle-plasma membrane fusion product (omega-profiles), secretion was halted at various time intervals after stimulation and the excess membrane allowed to transform into endocytotic vesicles in the presence of horseradish peroxidase. By adding horseradish peroxidase at various times after inhibition of secretion, the time course of membrane retrieval could be measured directly. All our results are consistent with a two-step kinetic model in which exocytosis and membrane retrieval are consecutive events. The estimated volumes of the compartments involved are roughly equal. The rate of endocytosis is strongly temperature-dependent but unaffected by extracellular calcium in the range of 10(-8)-2.5 X 10(-3) M, suggesting that calcium is not required at the site of endocytotic membrane fusion. Membrane retrieval is also unaffected by Lanthanum (1 mM) but is slowed by hypertonic media.


Effects of Vitamin D Supplementation on Muscle Strength in Athletes: A Systematic Review.
Chiang, C-m, Ismaeel, A, Griffis, RB, and Weems, S. Effects of vitamin D supplementation on muscle strength in athletes: A systematic review. J Strength Cond Res 31(2): 566-574, 2017-The purpose of this systematic review of the literature was to investigate the effects of vitamin D supplementation on muscle strength in athletes. A computerized literature search of 3 databases (PubMed, MEDLINE, and Scopus) was performed. Included in the review were randomized controlled trials (RCTs), published in English, which measured serum vitamin D concentrations and muscle strength in healthy, athletic participants aged 18-45 years. Quality was assessed using the PEDro scale. Five RCTs and 1 controlled trial were identified, and quality assessment showed 5 trials were of "excellent quality" and 1 was of "good quality." Trials lasted from 4 weeks to 6 months and dosages ranged from 600 to 5,000 International Units (IU) per day. Vitamin D2 was found to be ineffective at impacting muscle strength in both studies wherein it was administered. In contrast, vitamin D3 was shown to have a positive impact on muscle strength. In 2 studies, strength outcome measures were significantly improved after supplementation (p ≤ 0.05). In the other 2 studies administering vitamin D3, there were trends for improved muscle strength. Specifically, improvements in strength ranged from 1.37 to 18.75%. Additional studies are needed to confirm these associations.