The sex chromosomes of frogs: variability and tolerance offer clues to genome evolution and function.
Frog sex chromosomes offer an ideal system for advancing our understanding of genome evolution and function because of the variety of sex determination systems in the group, the diversity of sex chromosome maturation states, the ease of experimental manipulation during early development. After briefly reviewing sex chromosome biology generally, we focus on what is known about frog sex determination, sex chromosome evolution, and recent, genomics-facilitated advances in the field. In closing we highlight gaps in our current knowledge of frog sex chromosomes, and suggest priorities for future research that can advance broad knowledge of gene dose and sex chromosome evolution.


Association between MRI-defined osteoarthritis, pain, function and strength 3-10 years following knee joint injury in youth sport.
Youth and young adults who participate in sport have an increased risk of knee injury and subsequent osteoarthritis. Improved understanding of the relationship between structural and clinical outcomes postinjury could inform targeted osteoarthritis prevention interventions. This secondary analysis examines the association between MRI-defined osteoarthritis and self-reported and functional outcomes, 3-10 years following youth sport-related knee injury in comparison to healthy controls. Participants included a subsample (n=146) of the Alberta Youth Prevention of Early Osteoarthritis cohort: specifically, 73 individuals with 3-10years history of sport-related intra-articular knee injury and 73 age-matched, sex-matched and sport-matched controls with completed MRI studies. Outcomes included: MRI-defined osteoarthritis, radiographic osteoarthritis, Knee Injury and Osteoarthritis Outcome Score, Intermittent and Constant Osteoarthritis Pain, knee extensor/flexor strength, triple-hop and Y-balance test. Descriptive statistics and univariate logistic regression were used to compare those with and without MRI-defined osteoarthritis. Associations between MRI-defined osteoarthritis and each outcome were assessed using multivariable linear regression considering the influence of injury history, sex, body mass index and time since injury. Participant median age was 23 years (range 15-27), and 63% were female. MRI-defined osteoarthritis varied by injury history, injury type and surgical history and was not isolated to participants with ACL and/or meniscal injuries. Those with a previous knee injury had 10-fold (95% CI 2.3 to 42.8) greater odds of MRI-defined osteoarthritis than uninjured participants. MRI-defined osteoarthritis was independently significantly associated with quality of life, but not symptoms, strength or function. MRI-detected structural changes 3- 10 years following youth sport-related knee injury may not dictate clinical symptomatology, strength or function but may influence quality of life.


Guardianship and End-of-Life Decision Making.
As the population ages, more adults will develop impaired decision-making capacity and have no family members or friends available to make medical decisions on their behalf. In such situations, a professional guardian is often appointed by the court. This official has no preexisting relationship with the impaired individual but is paid to serve as a surrogate decision maker. When a professional guardian is faced with decisions concerning life-sustaining treatment, substituted judgment may be impossible, and reports have repeatedly suggested that guardians are reluctant to make the decision to limit care. Physicians are well positioned to assist guardians with these decisions and safeguard the rights of the vulnerable persons they represent. Doing so effectively requires knowledge of the laws governing end-of-life decisions by guardians. However, physicians are often uncertain about whether guardians are empowered to withhold treatment and when their decisions require judicial review. To address this issue, we analyzed state guardianship statutes and reviewed recent legal cases to characterize the authority of a guardian over choices about end-of-life treatment. We found that most state guardianship statutes have no language about end-of-life decisions. We identified 5 legal cases during the past decade that addressed a guardian's authority over these decisions, and only 1 case provided a broad framework applicable to clinical practice. Work to improve end-of-life decision making by guardians may benefit from a multidisciplinary effort to develop comprehensive standards to guide clinicians and guardians when treatment decisions need to be made.


Ubc9p and the conjugation of SUMO-1 to RanGAP1 and RanBP2.
The yeast UBC9 gene encodes a protein with homology to the E2 ubiquitin-conjugating enzymes that mediate the attachment of ubiquitin to substrate proteins [1]. Depletion of Ubc9p arrests cells in G2 or early M phase and stabilizes B-type cyclins [1]. p18(Ubc9), the Xenopus homolog of Ubc9p, associates specifically with p88(RanGAP1) and p340(RanBP2) [2]. Ran-binding protein 2 (p340(RanBP2)) is a nuclear pore protein [3] [4], and p88(RanGAP1) is a modified form of RanGAP1, a GTPase-activating protein for the small GTPase Ran [2]. It has recently been shown that mammalian RanGAP1 can be conjugated with SUMO-1, a small ubiquitin-related modifier [5-7], and that SUMO-1 conjugation promotes RanGAP1's interaction with RanBP2 [2,5,6]. Here we show that p18(Ubc9) acts as an E2-like enzyme for SUMO-1 conjugation, but not for ubiquitin conjugation. This suggests that the SUMO-1 conjugation pathway is biochemically similar to the ubiquitin conjugation pathway but uses a distinct set of enzymes and regulatory mechanisms. We also show that p18(Ubc9) interacts specifically with the internal repeat domain of RanBP2, which is a substrate for SUMO-1 conjugation in Xenopus egg extracts.


Is Motor Performance in 5.5-Year-Old Children Associated with the Presence of Generalized Joint Hypermobility?
To determine the prevalence of generalized joint hypermobility (GJH) in Dutch children aged 5.5 years, and to examine the association between GJH and motor performance and development over time. A prospective cohort of 249 children was recruited. GJH was assessed with the Beighton test at age 5.5 years. Motor performance was evaluated at age 2.0 years using the Bayley Scales of Infant Development, Second Edition and at age 5.5 years using the Movement Assessment Battery for Children-Second Edition (subscore categories: manual dexterity, aiming and catching, and static and dynamic balance). In 249 children, the prevalence of GJH, defined by the Beighton test score, was 34.1% for a score ≥ 4, 22.5% for a score ≥ 5, and 16.5% for a score ≥ 6. No significant association was found between GJH and total motor performance. Manual dexterity in girls (Beighton score ≥ 4) was positively associated with higher level of motor performance (β [SE] = 0.38 [0.17]; P = .028), ranging from +0.04 SD to +0.72 SD, even after correction for covariates. A significant interaction between GJH and body mass index (BMI) growth was found, indicating that the effect of GJH on the rate of development of motor performance declines with increasing BMI growth (β = 0.05 [0.02]; P = .031). In this healthy pediatric cohort, GJH was present in one-third of the sample, and no significant association was found between GJH and total motor performance. The effect of GJH on the rate of development of motor performance appears to decline with increasing BMI growth. Longitudinal prospective studies are recommended to detect influences of GJH on motor performance over time, as well as the influence of body composition and Beighton cutoff points.


Long-term dietary restriction differentially affects the expression of BDNF and its receptors in the cortex and hippocampus of middle-aged and aged male rats.
Dietary restriction (DR) exerts significant beneficial effects in terms of aging and age-related diseases in many organisms including humans. The present study aimed to examine the influence of long-term DR on the BDNF system at the transcriptional and translational levels in the cortex and hippocampus of middle-aged (12-month-old) and aged (24-month-old) male Wistar rats. The obtained results revealed that the DR upregulated the expression of exon-specific BDNF transcripts in both regions, followed by elevated levels of mBDNF only in the cortex in middle-aged animals. In aged animals, DR modulated BDNF protein levels by increasing proBDNF and by declining mBDNF levels. Additionally, elevated levels of the full-length TrkB accompanied by a decreased level of the less-glycosylated TrkB protein were observed in middle-aged rats following DR, while in aged rats, DR amplified only the expression of the less-glycosylated form of TrkB. The levels of phosphorylated TrkB(Y816) were stable during aging regardless of feeding. Reduced levels of p75(NTR) were detected in both regions of middle-aged DR-fed animals, while a significant increase was measured in the cortex of aged DR-fed rats. These findings shed additional light on DR as a modulator of BDNF system revealing its disparate effects in middle-aged and aged animals. Given the importance of the proBDNF/BDNF circuit-level expression in different brain functions and various aspects of behavior, it is necessary to further elucidate the optimal duration of the applied dietary regimen with regard to the animal age in order to achieve its most favorable effects.


Identification of priorities for improvement of medication safety in primary care: a PRIORITIZE study.
Medication error is a frequent, harmful and costly patient safety incident. Research to date has mostly focused on medication errors in hospitals. In this study, we aimed to identify the main causes of, and solutions to, medication error in primary care. We used a novel priority-setting method for identifying and ranking patient safety problems and solutions called PRIORITIZE. We invited 500 North West London primary care clinicians to complete an open-ended questionnaire to identify three main problems and solutions relating to medication error in primary care. 113 clinicians submitted responses, which we thematically synthesized into a composite list of 48 distinct problems and 45 solutions. A group of 57 clinicians randomly selected from the initial cohort scored these and an overall ranking was derived. The agreement between the clinicians' scores was presented using the average expert agreement (AEA). The study was conducted between September 2013 and November 2014. The top three problems were incomplete reconciliation of medication during patient 'hand-overs', inadequate patient education about their medication use and poor discharge summaries. The highest ranked solutions included development of a standardized discharge summary template, reduction of unnecessary prescribing, and minimisation of polypharmacy. Overall, better communication between the healthcare provider and patient, quality assurance approaches during medication prescribing and monitoring, and patient education on how to use their medication were considered the top priorities. The highest ranked suggestions received the strongest agreement among the clinicians, i.e. the highest AEA score. Clinicians identified a range of suggestions for better medication management, quality assurance procedures and patient education. According to clinicians, medication errors can be largely prevented with feasible and affordable interventions. PRIORITIZE is a new, convenient, systematic, and replicable method, and merits further exploration with a view to becoming a part of a routine preventative patient safety monitoring mechanism.


Altered lipid metabolism in parvovirus-infected cells.
A broad spectrum of cell lipid alterations are known to occur as a consequence of various viral infections. These changes include inhibition of lipid synthesis, stimulation of lipid synthesis and changes in the proportions of various lipids. The current study examined the effects of two parvoviruses on lipids of rat kidney (NRK) cells. Cells were infected with H-1 or Kilham rat virus (KRV) and the effects on 14C-acetate incorporation determined. Results showed that H-1 virus rapidly inhibited lipid formation (in 1 h) while KRV produced a similar effect beginning around 8 h. Pretreatment of the cells with cycloheximide did not alter this response. Fatty acid analysis by gas chromatography did not reveal major alterations in this component of total cell lipids although some fatty acids became undetectable by 18 h post-infection. The data suggest that these parvoviruses, especially H-1 virus, are able to rapidly alter lipid formation following infection and that this effect may be mediated by a virion component.


Paths of convergence for agriculture, health, and wealth.
This special feature calls for forward thinking around paths of convergence for agriculture, health, and wealth. Such convergence aims for a richer integration of smallholder farmers into national and global agricultural and food systems, health systems, value chains, and markets. The articles identify analytical innovation, where disciplines intersect, and cross-sectoral action where single, linear, and siloed approaches have traditionally dominated. The issues addressed are framed by three main themes: (i) lessons related to agricultural and food market growth since the 1960s; (ii) experiences related to the integration of smallholder agriculture into national and global business agendas; and (iii) insights into convergence-building institutional design and policy, including a review of complexity science methods that can inform such processes. In this introductory article, we first discuss the perspectives generated for more impactful policy and action when these three themes converge. We then push thematic boundaries to elaborate a roadmap for a broader, solution-oriented, and transdisciplinary approach to science, policies, and actions. As the global urban population crosses the 50% mark, both smallholder and nonsmallholder agriculture are keys in forging rural-urban links, where both farm and nonfarm activities contribute to sustainable nutrition security. The roadmaps would harness the power of business to reduce hunger and poverty for millions of families, contribute to a better alignment between human biology and modern lifestyles, and stem the spread of noncommunicable chronic diseases.


Donor bone marrow infusion in deceased and living donor renal transplantation.
The infusion and persistence in a transplant recipient of donor-derived bone marrow cells (DBMC) of multi-lineage can lead to a state of permanent chimerism. In solid vascular organ transplantation, the donor bone marrow lineage cells can even be derived from the transplant organ, and these cells can be detected in very small numbers in the recipient. This has been called microchimerism. Much controversy has developed with respect to the function of chimeric cells in organ transplantation. One idea is that the occurrence of these donor cells found in microchimerism in the recipient are coincidental and have no long-term beneficial effect on engraftment. A second and opposing view, is that these donor cells have immunoregulatory function that affect both the acute and chronic phases of the recipient anti-donor responses. It follows that detecting quantitative changes in chimerism might serve as an indication of the donor-specific alloimmune or regulatory response that could occur in concert with or independent of other adaptive immune responses. The latter, including autoimmune native disease, need to be controlled in the transplant organ. The safety and immune tolerance potential of DBMC infusion with deceased and living donor renal transplants was evaluated in a non-randomized trial at this center and compared with non-infused controls given identical immunosuppression. Overall DBMC infusions were well tolerated by the recipients. There were no complications from the infusion(s), no episodes of graft-vs-host disease (GVHD) and no increase infections or other complications. In the deceased DBMC-kidney trial, actuarial graft survival at 5 years was superior especially when graft survival was censored for recipient death. Acute rejections were significant reduced in patients given two DBMC infusions, and chronic rejection was dramatically reduced in all DBMC treated patients. The most interesting finding was that the degree of microchimerism slowly increased over the years the DBMC group that had exhibited no rejection episodes. In the DBMC-living related trial, the incidence of acute rejection did not differ between groups. However, DBMC chimerism in recipient iliac crest marrow had increased more rapidly than might be predicted from results previously seen in the cadaver group, despite four times fewer DBMC infused, with the generation of T- regulartory cells in-vitro assays.


Crosses between sexual and apomictic dandelions (Taraxacum). II. The breakdown of apomixis.
Some dandelions are diplosporous gametophytic apomicts. In order to study the inheritance and breakdown of apomixis, crosses were made between diploid sexuals and triploid apomicts. To investigate their breeding system, four nonapomictic diploid and 10 nonapomictic triploid hybrids were pollinated with diploids and the progenies were analysed. Seed fertility was significantly reduced in two diploid hybrids. Nine triploid hybrids were fertile and could be classified into three types, with respect to the composition of their progenies. Type A produced n+n hybrids. Type B produced either a mixture of n + n and 2n + n hybrids, or a mixture of pseudogamous 2n + 0 apomicts and 2n + n hybrids. Type C produced exclusively 2n + n hybrids. Inheritance of a microsatellite marker strongly suggested that 2n egg cells in type C plants were produced by a first division restitution mechanism. As in apomicts, microsporogenesis in type C plants was reductional. This suggests that type C plants are diplosporous plants that lack parthenogenesis. Such plants are very rare in other apomictic plant species. It is concluded that 'elements of apomixis', diplospory and parthenogenesis, can be uncoupled. This is inconsistent with the single-locus model for apomixis in Taraxacum as suggested by Mogie (1992). Instead, our results suggest that several loci are involved in the genetic control of apomixis in Taraxacum.


High-efficiency multipass Ti:sapphire amplifiers for a continuous-wave single-mode laser.
We present the amplification of a continuous-wave single-mode ring dye laser in Ti:sapphire. A peak gain of 2 x 10(6) has been obtained in a passive multipass amplifier, which yielded 20-nsec pulses of 0.7-mJ energy at 780 nm. We discuss the advantages of this passive multipass amplifier in comparison with a regenerative amplifier that we have also developed. By second-harmonic generation we obtained high-peak-power UV pulses from the amplified single-mode laser.


Perceptions of cardiovascular health in underserved communities.
Cardiovascular disease is the leading cause of deaths and illnesses in US adults, and the prevalence is disproportionately high in underserved populations. In this study, we assessed respondents' understanding of context-specific differences in knowledge and perceptions of disease, risk, and prevention in 6 underserved communities, with the longer-term goal of developing appropriate interventions. Thirty-nine small-group sessions and 14 interviews yielded data from 318 adults. Each site's researchers coded, analyzed, and extracted key themes from local data. Investigators from all sites synthesized results and identified common themes and differences. Themes clustered in 3 areas (barriers to cardiovascular health, constraints related to multiple roles, and suggestions for effective communications and programs). Barriers spanned individual, social and cultural, and environmental levels; women in particular cited multiple roles (eg, competing demands, lack of self-care). Programmatic suggestions included the following: personal, interactive, social context; information in language that people use; activities built around cultural values and interests; and community orientation. In addition, respondents preferred health-related information from trusted groups (eg, AARP), health care providers (but with noticeable differences of opinion), family and friends, and printed materials. Interventions to decrease barriers to cardiovascular health are needed; these strategies should include family and community context, small groups, interactive methods, culturally sensitive materials, and trusted information sources. New-immigrant communities need culturally and linguistically tailored education before receiving more substantive interventions.


Is there a place for serum laminin determination in patients with liver disease and cancer?
Laminin is a glycoprotein which has an important role in the mechanism of fibrogenesis and is, thus, related to hepatic fibrosis in addition to presenting increased levels in several types of neoplasias. However, its determination is not routinely considered in the study of hepatic fibrosis. In this review, the authors critically comment on the role of this glycoprotein compared to other markers of fibrosis through non-invasive procedures (Fibroscan). They also consider its clinical investigational potential and believe that the continuation of these investigations might contribute to a better understanding of the fibrogenic mechanism, which could in turn either lead to the identification of patients at risk of developing fibrosis non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) or at least be used as an indicator for hepatic biopsy in such patients. Finally, the authors believe that serum laminin determination might contribute to the diagnosis of epithelial tumor metastasis and peritoneal carcinomatosis.


Differentiation and growth of rat egg-cylinders cultured in vitro in a serum-free and protein-free medium.
Modified organ cultures of rat egg-cylinders were grown for 2 weeks in Eagle's minimal essential medium (MEM) without serum. Differentiation of epidermis and cartilage in the cultures deprived of serum was comparable to that in fully serum-supplemented medium, whereas other differentiated tissues were rare or absent. The purpose of the experiment was to determine whether terminal tissue differentiation is modified by various added factors. The factors used affected the growth and/or differentiation of explants as follows: bovine serum albumin and human transferrin had a positive permissive influence on the appearance of neuroblasts; human transferrin alone stimulated the formation of lentoids, a relatively rare tissue. Retinoic acid inhibited cartilage formation and stimulated the differentiation of cylindrical epithelium; neural growth factor inhibited the growth of explants; and 5-azacytidine impeded the survival of explants. One can conclude that these factors influenced the growth and differentiation of the early rat embryos cultured in a chemically defined medium.


Human intestinal sarcosporidiosis: report of six cases.
Specimens of resected small intestine from six patients aged 3 to 70 years with acute enteritis contained sexual forms of sarcosporidia. Histopathologically, the diagnoses were either segmental eosinophilic enteritis or segmental necrotizing enteritis. The presence of sarcosporidia in market beef (Bos indicus), and the patients' habit of eating the beef uncooked in the form of chili-hot dishes, suggest that the species is an ox-man parasite similar to Sarcocystis hominis (Railliet and Lucet, 1891) Dubey, 1976. Presence of numerous Gram-positive bacilli in segmental necrotizing enteritis suggests an interplay between two etiological agents in producing the hosts' inflammatory responses. Five patients recovered after resection, but one died due to extensive necrosis of the intestinal wall and leakage at the site of anastomosis. Only conventional antibiotics were given after the operations. None of the five surviving patients has had recurrent enteritis for at least 1 year.


Beat-frequency adjustable Er3+-doped DBR fiber laser for ultrasound detection.
A compact low beat-frequency dual-polarization distributed Bragg reflector (DBR) fiber laser whose beat frequency can be varied, for high-frequency ultrasound detection has been proposed and experimentally demonstrated. The laser was fabricated in small birefringent commercial erbium-doped fiber. It operated in a robust single-longitude mode with output power of more than 1 mW and high signal-to-noise ratio better than 60 dB. Induced birefringence to the fiber during the UV inscription process is small (~10(-7)) and consequently the laser beats at a low frequency of ~20 MHz which is at least one order of magnitude smaller than previously reported results, making frequency down-conversion unnecessary. The beat frequency can be adjusted by controlling the side-exposure time of the UV light irradiating the gain cavity, providing a simple approach to multiplex a large number of DBR fiber lasers of different frequencies in series using frequency division multiplexing (FDM) technique. The proposed DBR fiber laser is also temperature insensitive, making it a good candidate for hydrophone applications.


Integrated ultrafast saturable absorber.
An ultrafast saturable absorber was demonstrated experimentally in AlGaAs, operated with a photon energy below half the band gap, near 1555 nm. Both the saturation intensity and the linear transmission can be independently designed in the structure. The device is based on spatial soliton emission from a tapered channel waveguide with a corresponding increase in transmission of 50%. The experimental data compare favorably with numerically simulated results.


Selective recognition of the di/trimethylammonium motif by an artificial carboxycalixarene receptor.
Chemical tools that recognise post-translational modifications have promising applications in biochemistry and in therapy. We report a simple carboxycalixarene that selectively binds molecules containing di/trimethylammonium moieties in isolation, in cell lysates and when incorporated in histone peptides. Our findings reveal the potential of using carboxycalixarene-based receptors to study epigenetic regulation.


Delivering services to incarcerated teen fathers: a pilot intervention to increase the quality of father-infant interactions during visitation.
The absence of a father figure has been linked to very poor developmental outcomes for the child. During incarceration, there are limited opportunities for visitation between fathers and their children. The Baby Elmo Program provides incarcerated teen fathers with parenting training and visitation with their children with the stated goal of enhancing father-child interactional quality. Forty-one incarcerated teen fathers and their infants ranging from 1 to 15 months of age participated in the present study. During individual sessions, a trained facilitator prepared fathers for visits with their children by introducing key concepts such as following the child's lead, using developmentally appropriate media to illustrate those concepts. After each training session, the incarcerated teen father interacted with his infant and the visit was video recorded. Analysis of the visit sessions focused on father's time use on different activities, the quality of father-infant interactions, and father's integration of target skills introduced in the intervention. The time-use analysis revealed that time use changed as a function of infant age. Growth linear modeling indicated that there were significant positive increases in the amount of parent support and infant engagement as a function of the number of sessions. Follow-up analyses indicated that changes between specific sessions mapped onto the target skills discussed during specific training sessions. This study's preliminary findings suggest that an intervention integrating visitation and appropriate media may be effective for incarcerated teen fathers. Due to the lack of a randomized control group, the present findings are exploratory and are discussed with a focus on further program development.


The branchial chloride pump in the goldfish Carassius auratus: relationship between Cl-/HCO3- and Cl-/Cl-- exchanges and the effect of thiocyanate.
1. The effect of thiocyanate on chloride and sodium fluxes across the gill was studied in the goldfish Carassius auratus. At low external chloride concentrations, addition of SCN- to the bathing solution markedly inhibited chloride influx and efflux, the net flux being reversed, SCN- injection was without effect. SCN- had no effect on sodium fluxes when injected or added to the external medium. 2. The inhibition of chloride influx by SCN- was of a mixed type involving simultaneous modifications of the affinity constant of the carrier for Cl- and of the maximal Cl- influx. The affinity constant of the carrier for SCN- was 10 times lower than that for Cl-. 3. The gill of the goldfish was found to be practically impermeable to SCN-. 4. In the presence of external SCN-, the Cl-/HCO3- exchange was reversed: Cl- was lost against HCO3- which is absorbed. This suggests an obligatory exchange. 5. Exchange diffusion for chloride was also demonstrated. 6. A kinetic model is proposed to explain chloride and bicarbonate transport across the gill of Carassius auratus.


Minimally important difference estimates and methods: a protocol.
Patient-reported outcomes (PROs) are often the outcomes of greatest importance to patients. The minimally important difference (MID) provides a measure of the smallest change in the PRO that patients perceive as important. An anchor-based approach is the most appropriate method for MID determination. No study or database currently exists that provides all anchor-based MIDs associated with PRO instruments; nor are there any accepted standards for appraising the credibility of MID estimates. Our objectives are to complete a systematic survey of the literature to collect and characterise published anchor-based MIDs associated with PRO instruments used in evaluating the effects of interventions on chronic medical and psychiatric conditions and to assess their credibility. We will search MEDLINE, EMBASE and PsycINFO (1989 to present) to identify studies addressing methods to estimate anchor-based MIDs of target PRO instruments or reporting empirical ascertainment of anchor-based MIDs. Teams of two reviewers will screen titles and abstracts, review full texts of citations, and extract relevant data. On the basis of findings from studies addressing methods to estimate anchor-based MIDs, we will summarise the available methods and develop an instrument addressing the credibility of empirically ascertained MIDs. We will evaluate the credibility of all studies reporting on the empirical ascertainment of anchor-based MIDs using the credibility instrument, and assess the instrument's inter-rater reliability. We will separately present reports for adult and paediatric populations. No research ethics approval was required as we will be using aggregate data from published studies. Our work will summarise anchor-based methods available to establish MIDs, provide an instrument to assess the credibility of available MIDs, determine the reliability of that instrument, and provide a comprehensive compendium of published anchor-based MIDs associated with PRO instruments which will help improve the interpretability of outcome effects in systematic reviews and practice guidelines.


Structure and crystallization of SiO2 and B2O3 doped lithium disilicate glasses from theory and experiment.
Solid solutions of SiO2 and B2O3 in Li2O·2SiO2 are synthesized and characterized for the first time. Their structure and crystallization mechanisms are investigated employing a combination of simulations at the density functional theory level and experiments on the crystallization of SiO2 and B2O3 doped lithium disilicate glasses. The remarkable agreement of calculated and experimentally determined cell parameters reveals the preferential, kinetically controlled incorporation of [SiO4] and [BO4] at the Li+ lattice sites of the Li2O·2SiO2 crystal structure. While the addition of SiO2 increases the glass viscosity resulting in lower crystal growth velocities, glasses containing B2O3 show a reduction of both viscosities and crystal growth velocities. These observations could be rationalized by a change of the chemical composition of the glass matrix surrounding the precipitated crystal phase during the course of crystallization, which leads to a deceleration of the attachment of building units required for further crystal growth at the liquid-crystal interface.


Morphogen interpretation: concentration, time, competence, and signaling dynamics.
Tissue patterning during animal development is orchestrated by a handful of inductive signals. Most of these developmental cues act as morphogens, meaning they are locally produced secreted molecules that act at a distance to govern tissue patterning. The iterative use of the same signaling molecules in different developmental contexts demands that signal interpretation occurs in a highly context-dependent manner. Hence the interpretation of signal depends on the specific competence of the receiving cells. Moreover, it has become clear that the differential interpretation of morphogens depends not only on the level of signaling but also the signaling dynamics, particularly the duration of signaling. In this review, we outline molecular mechanisms proposed in recent studies that explain how the response to morphogens is determined by differential competence, pathway intrinsic feedback, and the interpretation of signaling dynamics by gene regulatory networks. WIREs Dev Biol 2017, 6:e271. doi: 10.1002/wdev.271 For further resources related to this article, please visit the WIREs website.


Apples and Oranges: Divergent Meanings of Parents' and Adolescents' Perceptions of Parental Influence.
This study examines the hypothesis that effective parental influence stems from the qualities of the parent-adolescent relationship rather than from explicit efforts to alter adolescents' behaviors. Adolescents' versus parents' perceptions of parental influence as predictors of parent-adolescent relationship quality and of adolescents' social functioning are examined using observational and multireporter data obtained from a sample of 167 adolescents (90 female, 77 male; age M = 13.34 years, SD = 0.65), their parents, and their same-sex peers. Analyses revealed that adolescents' and parents' perceptions of parental influence were uncorrelated with one another and were differentially related to qualities of adolescents' relationships with parents and friends. Adolescents' perceptions of high parental influence were linked to observations and self-reports of warm, supportive relationships with parents (particularly mothers). In contrast, parents' reports of high influence were linked to lower levels of adolescent autonomy with parents and friends and less relatedness with mothers and friends.


Wave-making by whirligig beetles (gyrinidae).
Swimming whirligig beetles (Dineutes carolinus) either make no waves at all or make conspicuous circular or vee-shaped patterns of capillary waves. The beetle's swimming speed can be determined from these wave patterns (or lack of them). Capillary waves precede the beetle for several body lengths, and their reflections may help the beetle avoid solid objects by echolocation. The gravity waves produced by a beetle are always longer than the beetle's hull length. Hence the waves do not interact with the hull to impose an upper limit on speed as they do with conventional ships. Although the beetles swim at high speeds, they apparently do not hydroplane.


Perioperative high-dose-rate brachytherapy (PHDRB) in previously irradiated head and neck cancer: Initial results of a Phase I/II reirradiation study.
This study was undertaken to determine the feasibility of salvage surgery and perioperative high-dose-rate brachytherapy (PHDRB) at the dose/fractionation schedule proposed in patients with previously irradiated, recurrent head and neck cancer or second primary tumors arising in a previously irradiated field. Twenty-five patients were treated with surgical resection and PHDRB. The PHDRB dose was 4 Gy b.i.d. x 8 (32 Gy) for R0 resections and 4 Gy b.i.d. x 10 (40 Gy) for R1 resections. Further external beam radiotherapy or chemotherapy was not given. Resections were categorized as R0 (negative margins of at least 10 mm) in 3 patients (12.0%) and R1 (negative margins of less than 10 mm or microscopically positive margins) in 22 (88.0%). Twelve patients with R1 resections had microscopically positive margins (48%), and 10 patients had close margins (40%), with a median of 2.0 mm. Ten patients (40.0%) developed Radiation Therapy Oncology Group Grade 3 or greater toxicity. Seven patients (28%) presented complications requiring a major surgical procedure. Four of these complications appeared in the immediate postoperative period and were surgical in nature (flap failure, n = 2; fistula, n = 2), and the other three were mainly related to the brachytherapy procedure (n = 2) or the radiation dose delivered (n = 1). One patient died on postoperative day 11 due to bleeding. After a median followup of 14 months, the 4-year local control rate and overall survival were 85.6% and 46.4%, respectively. Surgical salvage and PHDRB at the dose/fractionation proposed are feasible in this high-risk population. Toxicity is high, but not substantially different from other reirradiation series. Four-year local control results are encouraging taking into account that 22 of 25 patients (88%) had either close or microscopically positive margins.


Profiling in basketball: physical and physiological characteristics of elite players.
The purpose of this study was to describe structural and functional characteristics of elite Serbian basketball players and to evaluate whether players in different positional roles have different physical and physiological profiles. Five men's basketball teams participated in the study and competed in the professional First National League. Physiological measurements were taken of 60 players during the final week of their preparatory training for competition. According to positional roles, players were categorized as guards (n = 20), forwards (n = 20), and centers (n = 20). Guards were older (p < 0.01) and more experienced (p < 0.01) as compared with both forwards and centers. Centers were taller and heavier than guards and forwards (p < 0.01), whereas forwards had significantly higher height and weight than guards (p < 0.01). Centers had more body fat (p < 0.01) as compared with forwards and guards. Also, centers had significantly lower estimated VO(2)max values (p < 0.01) compared with forwards and guards. In addition, the highest heart rate frequencies during the last minute of the shuttle run test were lower in guards (p < 0.01) as compared with forwards and centers. Vertical jump power was significantly higher in centers (p < 0.01) as compared with guards. The results of the present study demonstrate that a strong relationship exists between body composition, aerobic fitness, anaerobic power, and positional roles in elite basketball.


Dichotomous "Good Outcome" Indicates Mobility More Than Cognitive or Social Quality of Life.
Worthwhile interventions for intracerebral hemorrhage or subarachnoid hemorrhage generally hinge on whether they improve the odds of good outcome. Although good outcome is correlated with mobility, correlations with other domains of health-related quality of life, such as cognitive function and social functioning, are not well described. We tested the hypothesis that good outcome is more closely associated with mobility than other domains. We defined "good outcome" as 0 through 3 (independent ambulation or better) versus 4 through 5 (dependent) on the modified Rankin Scale at 1, 3, and 12 months. We simultaneously assessed the modified Rankin Scale and health-related quality of life using web-based computer adaptive testing in the domains of mobility, cognitive function (executive function and general concerns), and satisfaction with social roles and activities. We compared the area under the curve between different health-related quality of life domains. Neurologic ICU with web-based follow-up. One hundred fourteen patients with subarachnoid hemorrhage or intracerebral hemorrhage. None. We longitudinally followed 114 survivors with data at 1 month, 62 patients at 3 months, and 58 patients at 12 months. At 1 month, area under the curve was highest for mobility (0.957; 95% CI, 0.904-0.98), higher than cognitive function-general concerns (0.819; 95% CI, 0.715-0.888; p = 0.003 compared with mobility), satisfaction with social roles and activities (0.85; 95% CI, 0.753-0.911; p = 0.01 compared with mobility), and cognitive function-executive function (0.879; 95% CI, 0.782-0.935; p = 0.058 compared with mobility). Optimal specificity and sensitivity for receiver operating characteristic analysis were approximately 1.5 SD below the U.S. population mean. Health-related quality of life assessments reliably distinguished between good and poor outcomes as determined by the modified Rankin Scale. Good outcome indicated health-related quality of life about 1.5 SD below the U.S. population mean. Associations were weaker for cognitive function and social function than mobility.


Electrophysiologic testing to identify patients with coronary artery disease who are at risk for sudden death. Multicenter Unsustained Tachycardia Trial Investigators.
The mortality rate among patients with coronary artery disease, abnormal ventricular function, and unsustained ventricular tachycardia is high. The usefulness of electrophysiologic testing for risk stratification in these patients is unclear. We performed electrophysiologic testing in patients who had coronary artery disease, a left ventricular ejection fraction of 40 percent or less, and asymptomatic, unsustained ventricular tachycardia. Patients in whom sustained ventricular tachyarrhythmias could be induced were randomly assigned to receive either antiarrhythmic therapy guided by electrophysiologic testing or no antiarrhythmic therapy. The primary end point was cardiac arrest or death from arrhythmia. Patients without inducible tachyarrhythmias were followed in a registry. We compared the outcomes of 1397 patients in the registry with those of 353 patients with inducible tachyarrhythmias who were randomly assigned to receive no antiarrhythmic therapy in order to assess the prognostic value of electrophysiologic testing. Patients were followed for a median of 39 months. In a Kaplan-Meier analysis, two-year and five-year rates of cardiac arrest or death due to arrhythmia were 12 and 24 percent, respectively, among the patients in the registry, as compared with 18 and 32 percent among the patients with inducible tachyarrhythmias who were assigned to no antiarrhythmic therapy (adjusted P<0.001). Overall mortality after five years was 48 percent among the patients with inducible tachyarrhythmias, as compared with 44 percent among the patients in the registry (adjusted P=0.005). Deaths among patients without inducible tachyarrhythmias were less likely to be classified as due to arrhythmia than those among patients with inducible tachyarrhythmias (45 and 54 percent, respectively; P=0.06). Patients with coronary artery disease, left ventricular dysfunction, and asymptomatic, unsustained ventricular tachycardia in whom sustained ventricular tachyarrhythmias cannot be induced have a significantly lower risk of sudden death or cardiac arrest and lower overall mortality than similar patients with inducible sustained tachyarrhythmias.


When the SWI/SNF complex remodels...the cell cycle.
Mammalian cells contain several chromatin-remodeling complexes associated with the Brm and Brg1 helicase-like proteins. These complexes likely represent the functional homologs of the SWI/SNF and RSC complexes found in Saccharomyces cerevisiae. The mammalian chromatin-remodeling complexes are involved in both activation and repression of a variety of genes. Several lines of evidence also indicate that they play a specific role in the regulation of cell growth. Brm is down-regulated by ras signaling and its forced re-expression suppresses transformation by this oncogene. Besides, the Brg1 gene is silenced or mutated in several tumors cell lines and a Brg1-associated complex was recently found to co-purify with BRCA1, involved in breast and ovarian cancers. Finally, the gene encoding SNF5/Ini1, a subunit common to all mammalian SWI/SNF complexes, is inactivated in rhabdoid sarcomas, a very aggressive form of pediatric cancer. The current review will address observations made upon inactivation of Brm, Brg1 and SNF5/Ini1 by homologous recombination in the mouse, as well as the possible implication of these factors in the regulation of the Retinoblastoma pRb-mediated repression of the transcription factor E2F.


Acute Stress Promotes Aggressive-Like Behavior in Rats Made Allergic to Tree Pollen.
It has been reported that allergies are associated with depression and possibly suicide in women. Aggression is an important behavioral component that predisposes depressed individuals to suicidal acts. In the present study we examined the relationship between allergies and aggression to determine a potential contribution of allergies in factors of risk for suicidal behavior. Because stress plays a critical role in the manifestation of clinical symptoms of allergies and also in suicidal behavior, we also studied the role of acute stress. Female inbred Brown Norway rats known for their susceptibility to respiratory allergies were sensitized and challenged with a mixture of tree pollen and evaluated in the resident-intruder test for detection of aggressive behaviors. They were also subjected to acute stress by sessions of inescapable forced swimming and re-evaluated in the resident intruder test. Animals made allergic to tree pollen and subjected to acute stress displayed increased aggressive-like behavior as compared with control-saline treated animals or to their own aggressive scores previous to the stress session. These results suggest that allergies and stress increases aggressive-like behavior, indicating that these conditions may be important factors promoting altered emotional reactivity with the potential to influence suicidal behavior.


RB and p53 cooperate to prevent liver tumorigenesis in response to tissue damage.
The tumor suppressors retinoblastoma (RB) and p53 are important regulators of the cell cycle. Although human cancer cells inactivate RB and p53 by many mechanisms, the cooperative roles of these proteins in tumorigenesis are complex and tissue specific. We analyzed the cooperation of RB and p53 in liver development and pathogenesis of hepatocellular carcinoma. Spontaneous and carcinogen-induced (diethylnitrosamine) tumorigenesis were studied in mice with liver-specific deletions of Rb and/or p53 (Rbf/f;albcre+, p53f/f;albcre+ and Rbf/f; p53f/f;albcre+ mice). Genotype, histologic, immunohistochemical, microarray, quantitative polymerase chain reaction, immunoblot, and comparative genomic hybridization analyses were performed using normal and tumor samples. Comparative microarray analyses were performed against publicly available human microarray data sets. Deletion of RB and p53 from livers of mice deregulated the transcriptional programs associated with human disease. These changes were not sufficient for spontaneous tumorigenesis; potent quiescence mechanisms compensated for loss of these tumor suppressors. In response to hepatocarcinogen-induced damage, distinct and cooperative roles of RB and p53 were revealed; their loss affected cell cycle control, checkpoint response, and genome stability. In damaged tissue, combined loss of RB and p53 resulted in early lesion formation, aggressive tumor progression, and gene expression signatures and histologic characteristics of advanced human hepatocellular carcinoma. The effects RB and p53 loss are determined by the tissue environment; cell stresses that promote aggressive disease reveal the functions of these tumor suppressors.


Perioperative analgesia: a factor in the development of heel pressure ulcers?
This article presents the incidence of heel pressure ulcers after an elective hip or a knee replacement. The majority of patients in the authors' institute receive either a neuraxial block (epidural and/or spinal) or peripheral nerve blocks (femoral and sciatic), depending on the anaesthetist's and surgeon's preference, and the patient's physical status. The past few years have seen increasing use of peripheral nerve blocks for knee replacement surgery. Patients with either the central or peripheral nerve blockade are at an increased risk of developing heel pressure ulcers. This article describes the experience at a tertiary referral centre and, in particular, highlights the risk of developing heel ulcers in patients receiving peripheral nerve blocks. Medical and nursing staff looking after these patients should be made aware of this complication and appropriate measures should be taken to prevent this avoidable complication.


Design, synthesis and pharmacological evaluation of new 2-oxo-quinoline derivatives containing α-aminophosphonates as potential antitumor agents.
A series of novel 2-oxo-quinoline derivatives containing α-aminophosphonates were designed and synthesized as antitumor agents. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay results demonstrated that some compounds exhibited moderate to high inhibitory activity against HepG2, SK-OV-3 and NCI-H460 tumor cell lines, and most compounds showed much lower cytotoxicity against HL-7702 normal cells than 5-FU and cisplatin. The action mechanism of representative compound 5b was investigated by fluorescence staining assay, flow cytometric analysis and western blot (WB) assay, which indicated that this compound induced apoptosis and G2/M phase arrest accompanied by an increase in the production of intracellular Ca2+ and reactive oxygen species (ROS) and affecting associated enzymes and genes.


Study of Sleep Disorders among Young Children Using Hindi Translated and Validated Version of Pediatric Sleep Questionnaire.
The objective of this study is to assess the prevalence of sleep disorders among children aging between 4 and 9 years using Hindi version of Pediatric Sleep Questionnaire (PSQ). This study had two parts first, translation and validation of PSQ into Hindi language, and second, assessment of the prevalence of sleep disorders using PSQ Hindi version. Hindi PSQ was distributed in randomly chosen primary schools in a semi-urban area. The children were requested to get them filled by their parents. When the questionnaires were returned, responses were analyzed. Most of the items of the Hindi version had perfect agreement with original questionnaire in a bilingual population (κ =1). Totally, 435 children were included in the field study having average age of 6.3 years. Obstructive sleep apnea was reported in 7.5% children; symptoms suggestive of restless legs syndrome were reported by 2%-3%; teeth grinding by 13.9% and sleep talking by 22.6% children. PSQ Hindi version is a validated tool to screen for sleep disorders among children. Sleep disorders are fairly prevalent among young children in India.


ProQ3D: improved model quality assessments using deep learning.
Protein quality assessment is a long-standing problem in bioinformatics. For more than a decade we have developed state-of-art predictors by carefully selecting and optimising inputs to a machine learning method. The correlation has increased from 0.60 in ProQ to 0.81 in ProQ2 and 0.85 in ProQ3 mainly by adding a large set of carefully tuned descriptions of a protein. Here, we show that a substantial improvement can be obtained using exactly the same inputs as in ProQ2 or ProQ3 but replacing the support vector machine by a deep neural network. This improves the Pearson correlation to 0.90 (0.85 using ProQ2 input features). ProQ3D is freely available both as a webserver and a stand-alone program at http://proq3.bioinfo.se/. firstname.lastname@example.com. Supplementary data are available at Bioinformatics online.


Subchondral bone marrow lesions are highly associated with, and predict subchondral bone attrition longitudinally: the MOST study.
Subchondral bone attrition (SBA) is defined as flattening or depression of the osseous articular surface. The causes of attrition are unknown, but remodeling processes due to chronic overload that are reflected as bone marrow edema-like lesions (BMLs) on magnetic resonance imaging (MRI) might predispose the subchondral bone to subsequent attrition. The aim of this study was to evaluate the cross-sectional and longitudinal association of BMLs with SBA in the same subregion of the knee. The Multicenter Osteoarthritis (MOST) study is a longitudinal observational study of individuals who have or are at high risk for knee osteoarthritis. Subjects with available baseline and 30-months follow-up MRI were included. Patients with a recent history of trauma or findings suggestive of post-traumatic bone marrow changes were excluded. Subchondral BMLs and SBA were scored semiquantitatively from 0 to 3 in 10 tibiofemoral subregions. We evaluated the association of prevalent BMLs at baseline with the presence of prevalent and incident SBA on a per-subregion basis using logistic regression. We also cross-sectionally evaluated the association of BML grade severity and presence of baseline SBA. One thousand and twenty-five knees were included. 8.9% of the analyzed knee subregions showed SBA present at baseline and 9.2% of subregions exhibited prevalent subchondral BMLs. The adjusted odds ratio (OR) for prevalent SBA for subregions with prevalent BMLs was 18.8 [95% confidence intervals (CI) 15.9-22.4]. A larger BML size was directly associated with an increased risk of prevalent SBA. 195 (2.2%) subregions exhibited incident SBA at follow-up. The adjusted OR for incident SBA was 5.3 [95% CI 3.6-7.7] when compared to subregions without BMLs as the reference. Prevalent and incident SBA is strongly associated with subchondral BMLs in the same subregion. One explanation for the presence and development of SBA is subchondral remodeling due to increased stress, which is reflected as BMLs on MRI.


Multi-cellular 3D human primary liver cell culture elevates metabolic activity under fluidic flow.
We have developed a low-cost liver cell culture device that creates fluidic flow over a 3D primary liver cell culture that consists of multiple liver cell types, including hepatocytes and non-parenchymal cells (fibroblasts, stellate cells, and Kupffer cells). We tested the performance of the cell culture under fluidic flow for 14 days, finding that hepatocytes produced albumin and urea at elevated levels compared to static cultures. Hepatocytes also responded with induction of P450 (CYP1A1 and CYP3A4) enzyme activity when challenged with P450 inducers, although we did not find significant differences between static and fluidic cultures. Non-parenchymal cells were similarly responsive, producing interleukin 8 (IL-8) when challenged with 10 μM bacterial lipoprotein (LPS). To create the fluidic flow in an inexpensive manner, we used a rocking platform that tilts the cell culture devices at angles between ±12°, resulting in a periodically changing hydrostatic pressure drop between reservoirs and the accompanying periodically changing fluidic flow (average flow rate of 650 μL min(-1), and a maximum shear stress of 0.64 dyne cm(-2)). The increase in metabolic activity is consistent with the hypothesis that, similar to unidirectional fluidic flow, primary liver cell cultures increase their metabolic activity in response to fluidic flow periodically changes direction. Since fluidic flow that changes direction periodically drastically changes the behavior of other cells types that are shear sensitive, our findings support the theory that the increase in hepatic metabolic activity associated with fluidic flow is either activated by mechanisms other than shear sensing (for example increased opportunities for gas and metabolite exchange), or that it follows a shear sensing mechanism that does not depend on the direction of shear. Our mode of device operation allows us to evaluate drugs under fluidic cell culture conditions and at low device manufacturing and operation costs.


Gene electrotransfer to skin; review of existing literature and clinical perspectives.
Gene electrotransfer, which designates the combination of gene transfer and electroporation, is a non-viral means for transfecting genes into cells and tissues. It is a safe and efficient method and reports regarding the use of this technique in a variety of animal models and organs have been published in the literature. We find that gene electrotransfer to skin is of particular interest; not only due to the easy accessibility of this organ, which renders both treatment and evaluation feasible, but also the capability of the skin to produce transgenes and elicit immunological responses. Up to now more than 40 papers have been published in which gene electrotransfer was the technique used for gene transfection to skin in vivo. The aim of this review is to summarize which plasmids were injected and the electrical parameters applied. Furthermore an overview of the clinical perspectives of gene electrotransfer to skin will be presented.


Impact of physical activity category on incidence of cardiovascular disease: Results from the 10-year follow-up of the ATTICA Study (2002-2012).
The aim of the study was to examine the effects of physical activity (PA) level on 10-year cardiovascular disease (CVD) incidence, taking into consideration several clinical and lifestyle risk factors along with the potential moderating role of gender. An analysis was undertaken on data from the ATTICA prospective cohort study (10-year follow-up, 2002-2012), which followed a Greek adult population (aged 18-89years). A total n=317 of fatal and nonfatal CVD events occurred among the 2020 participants. After adjusting for the lifestyle and clinical risk factors as potential confounders, odds ratio (ORs) of CVD risk of individuals who reported being sufficiently active and highly active were decreased by 58% (95% CI: 0.30, 0.58) and 70% (95% CI: 0.15, 0.56), when compared to those who were inactive/insufficiently active, respectively. Men had nearly two-fold increase in risk of CVD (95% CI: 1.62, 2.18) versus women. Stratified analysis by gender, revealed that sufficiently active men, had 52% (95% CI: 0.24, 0.97) reduced risk of CVD incidence when compared to inactive males, while, for women, the role of PA lost significance following adjusting for lifestyle factors. The current data suggests a beneficial effect of even moderate physical activity levels on 10-year incidence of CVD, reinforcing the importance of physically activity, especially for men.


The UDP-glucose ceramide glycosyltransferase (UGCG) and the link to multidrug resistance protein 1 (MDR1).
The UDP-glucose ceramide glycosyltransferase (UGCG) is a key enzyme in the sphingolipid metabolism by generating glucosylceramide (GlcCer), the precursor for all glycosphingolipids (GSL), which are essential for proper cell function. Interestingly, the UGCG is also overexpressed in several cancer types and correlates with multidrug resistance protein 1 (MDR1) gene expression. This membrane protein is responsible for efflux of toxic substances and protects cancer cells from cell damage through chemotherapeutic agents. Studies showed a connection between UGCG and MDR1 overexpression and multidrug resistance development, but the precise underlying mechanisms are unknown. Here, we give an overview about the UGCG and its connection to MDR1 in multidrug resistant cells. Furthermore, we focus on UGCG transcriptional regulation, the impact of UGCG on cellular signaling pathways and the effect of UGCG and MDR1 on the lipid composition of membranes and how this could influence multidrug resistance development. To our knowledge, this is the first review presenting an overview about UGCG with focus on the relationship to MDR1 in the process of multidrug resistance development.


Transcriptome characterization by RNA-seq unravels the mechanisms of butyrate-induced epigenomic regulation in bovine cells.
Short-chain fatty acids (SCFAs), especially butyrate, affect cell differentiation, proliferation, and motility. Butyrate also induces cell cycle arrest and apoptosis through its inhibition of histone deacetylases (HDACs). In addition, butyrate is a potent inducer of histone hyper-acetylation in cells. Therefore, this SCFA provides an excellent in vitro model for studying the epigenomic regulation of gene expression induced by histone acetylation. In this study, we analyzed the differential in vitro expression of genes induced by butyrate in bovine epithelial cells by using deep RNA-sequencing technology (RNA-seq). The number of sequences read, ranging from 57,303,693 to 78,933,744, were generated per sample. Approximately 11,408 genes were significantly impacted by butyrate, with a false discovery rate (FDR) <0.05. The predominant cellular processes affected by butyrate included cell morphological changes, cell cycle arrest, and apoptosis. Our results provided insight into the transcriptome alterations induced by butyrate, which will undoubtedly facilitate our understanding of the molecular mechanisms underlying butyrate-induced epigenomic regulation in bovine cells.


Distinct cellular and subcellular patterns of expression imply distinct functions for the Drosophila homologues of moesin and the neurofibromatosis 2 tumor suppressor, merlin.
Interest in members of the protein 4.1 super-family, which includes the ezrin-radixin-moesin (ERM) group, has been stimulated recently by the discovery that the human neurofibromatosis 2 (NF2) tumor suppressor gene encodes an ERM-like protein, merlin. Although many proteins in this family are thought to act by linking the actin-based cytoskeleton to transmembrane proteins, the cellular functions of merlin have not been defined. To investigate the cellular and developmental functions of these proteins, we have identified and characterized Drosophila homologues of moesin (Dmoesin) and of the NF2 tumor suppressor merlin (Dmerlin). Using specific antibodies, we show that although these proteins are frequently coexpressed in developing tissues, they display distinct subcellular localizations. While Dmoesin is observed in continuous association with the plasma membrane, as is typical for an ERM family protein, Dmerlin is found in punctuate structures at the membrane and in the cytoplasm. Investigation of Dmerlin cultured cells demonstrates that it is associated with endocytic compartments. As a result of these studies, we propose that the merlin protein has unique functions in the cell which differ from those of other ERM family members.


Chronic thromboembolic pulmonary hypertension: evolution in management.
Chronic thromboembolic pulmonary hypertension (CTEPH) is an important cause of pulmonary hypertension. Although surgery is potentially curative, some patients present with inoperable disease. In these patients, medical therapies for pulmonary arterial hypertension are increasingly being used. The pathobiology of CTEPH development remains incompletely understood; however, evidence supports both large and small vessel disorder in patients with the disease. Surgical thromboendarterectomy is an increasingly well tolerated and often curative procedure and is the management strategy of choice for most patients. Although excellent outcomes in surgical management have been noted, the role of medical management in selected patients with inoperable or recurrent or persistent disease after surgery is increasing. A recent large, randomized controlled clinical trial of riociguat in CTEPH demonstrated improvements in exercise capacity, functional class, and hemodynamics. A safe, effective angioplasty approach to CTEPH is being pursued in addition. The approach to CTEPH management in the operable patient remains surgical, without clear benefit to preoperative pulmonary arterial hypertension-specific therapy at this time. Patients with inoperable disease or pulmonary hypertension following thromboendarterectomy, however, should be considered for medical management, with riociguat currently having the strongest evidence specific to CTEPH.


Homecoming of Citizen Soldiers: Postdeployment Problems and Service Use Among Army National Guard Soldiers.
The present study described the types and amount of problems and services sought among returned deployed Army National Guard soldiers (4568 soldiers in 50 units). The study responds to gaps in the research literature to better understand community intervention needs of reservists. About half (48%) of the soldiers reported one or two problems, mostly those of psychological well-being, such as feelings of anger and frustration, upsetting memories, and troubled sleep (34% of the study sample), followed by problems of social support (18%), alcohol use (17%), feelings of isolation including suicidal thoughts (13%), and financial difficulties (11%). Having engaged in direct combat and having wounded or killed someone showed positive relationships with reported problems. One-third (35%) of soldiers who reported having used services went to one service and, generally, soldiers went to services related to their expressed problems. Variance in self-reported problems explained by service use was low, suggesting unsought postdeployment services.


RNA sequencing: advances, challenges and opportunities.
In the few years since its initial application, massively parallel cDNA sequencing, or RNA-seq, has allowed many advances in the characterization and quantification of transcriptomes. Recently, several developments in RNA-seq methods have provided an even more complete characterization of RNA transcripts. These developments include improvements in transcription start site mapping, strand-specific measurements, gene fusion detection, small RNA characterization and detection of alternative splicing events. Ongoing developments promise further advances in the application of RNA-seq, particularly direct RNA sequencing and approaches that allow RNA quantification from very small amounts of cellular materials.


Detection and strain differentiation of infectious bronchitis virus in tracheal tissues from experimentally infected chickens by reverse transcription-polymerase chain reaction. Comparison with an immunohistochemical technique.
Oligonucleotide pairs were constructed for priming the amplification of fragments of nucleocapsid (N) protein and spike glycoprotein (S) genes of avian infectious bronchitis virus (IBV) by reverse transcriptionpolymerase chain reaction (RT-PCR). One oligonucleotide pair amplified a common segment of the N-gene and could detect various strains of IBV in allantoic fluid from inoculated chicken embryos, and in tracheal tissue preparations from experimentally infected chickens. Four pairs of oligonucleotides selectively primed the amplification of the S1 gene of Massachusetts/Connecticut, D1466, D274/D3896 and 793B strains of IBV, respectively. Groups of specific pathogen free chickens were experimentally inoculated with the Massachusetts (H120, M41), the D1466 and the 793B strains of IBV, and tracheal tissue preparations were made from each bird for RT-PCR and for immunohistochemistry (IHC) up to 3 days post-inoculation. The N-gene RT-PCR detected IBV in 82% of the chickens, while IHC only detected IBV in 60%. This difference was significant (P<0.02). The detection rate by N-gene RT-PCR varied from 67 to 100% for the various strains of IBV inoculated. The S1 gene oligonucleotide pairs were applied to the same tissue preparations and they detected specifically the Massachusetts (M41 and H120), the D1466 and the 793B strains of IBV at rates varying between 58 and 92%. When the mixtures of the primers were applied, the detection rate in tissue preparations was reduced to the level of 50 to 67%. It is concluded that the direct detection of IBV in tracheal tissues by RT-PCR is more sensitive than IHC and that the RT-PCR technique is able to distinguish between types of IBV.


Exposure to an Extremely-Low-Frequency Magnetic Field Stimulates Adrenal Steroidogenesis via Inhibition of Phosphodiesterase Activity in a Mouse Adrenal Cell Line.
Extremely low-frequency magnetic fields (ELF-MFs) are generated by power lines and household electrical devices. In the last several decades, some evidence has shown an association between ELF-MF exposure and depression and/or anxiety in epidemiological and animal studies. The mechanism underlying ELF-MF-induced depression is considered to involve adrenal steroidogenesis, which is triggered by ELF-MF exposure. However, how ELF-MFs stimulate adrenal steroidogenesis is controversial. In the current study, we investigated the effect of ELF-MF exposure on the mouse adrenal cortex-derived Y-1 cell line and the human adrenal cortex-derived H295R cell line to clarify whether the ELF-MF stimulates adrenal steroidogenesis directly. ELF-MF exposure was found to significantly stimulate adrenal steroidogenesis (p < 0.01-0.05) and the expression of adrenal steroid synthetic enzymes (p < 0.05) in Y-1 cells, but the effect was weak in H295R cells. Y-1 cells exposed to an ELF-MF showed significant decreases in phosphodiesterase activity (p < 0.05) and intracellular Ca2+ concentration (p < 0.01) and significant increases in intracellular cyclic adenosine monophosphate (cAMP) concentration (p < 0.001-0.05) and cAMP response element-binding protein phosphorylation (p < 0.05). The increase in cAMP was not inhibited by treatment with NF449, an inhibitor of the Gs alpha subunit of G protein. Our results suggest that ELF-MF exposure stimulates adrenal steroidogenesis via an increase in intracellular cAMP caused by the inhibition of phosphodiesterase activity in Y-1 cells. The same mechanism may trigger the increase in adrenal steroid secretion in mice observed in our previous study.


Predictors of phylogeographic structure among codistributed taxa across the complex Australian monsoonal tropics.
Differences in the geographic scale and depth of phylogeographic structure across codistributed taxa can reveal how microevolutionary processes such as population isolation and persistence drive diversification. In turn, environmental heterogeneity, species' traits, and historical biogeographic barriers may influence the potential for isolation and persistence. Using extensive SNP data and a combination of population genetic summary statistics and landscape genomic analyses, we explored predictors of the scale and depth of phylogeographic structure in codistributed lizard taxa from the topographically and climatically complex monsoonal tropics (AMT) of Australia. We first resolved intraspecific lineages and then tested whether genetic divergence across space within lineages is related to isolation by distance, resistance and/or environment and whether these factors differ across genera or between rock-related versus habitat generalist taxa. We then tested whether microevolutionary processes within lineages explain differences in the geographic scale and depth of intraspecific phylogeographic lineages. The results indicated that landscape predictors of phylogeographic structure differ between taxa. Within lineages, there was prevalent isolation by distance, but the strength of isolation by distance is independent of the taxonomic family, habitat specialization, and climate. Isolation by environment is the strongest predictor of landscape-scale genetic divergence for all taxa, with both temperature and precipitation acting as limiting factors. The strength of isolation by distance does not predict the geographic scale of the phylogeographic structure. However, more localized lineages had higher mean individual heterozygosity and less negative Tajima's D. This result implies that finer-scale phylogeographic structuring within species is associated with larger and more stable populations and, hence, persistence.


Effect of an indwelling pleural catheter vs chest tube and talc pleurodesis for relieving dyspnea in patients with malignant pleural effusion: the TIME2 randomized controlled trial.
Malignant pleural effusion causes disabling dyspnea in patients with a short life expectancy. Palliation is achieved by fluid drainage, but the most effective first-line method has not been determined. To determine whether indwelling pleural catheters (IPCs) are more effective than chest tube and talc slurry pleurodesis (talc) at relieving dyspnea. Unblinded randomized controlled trial (Second Therapeutic Intervention in Malignant Effusion Trial [TIME2]) comparing IPC and talc (1:1) for which 106 patients with malignant pleural effusion who had not previously undergone pleurodesis were recruited from 143 patients who were treated at 7 UK hospitals. Patients were screened from April 2007-February 2011 and were followed up for a year. Indwelling pleural catheters were inserted on an outpatient basis, followed by initial large volume drainage, education, and subsequent home drainage. The talc group were admitted for chest tube insertion and talc for slurry pleurodesis. Patients completed daily 100-mm line visual analog scale (VAS) of dyspnea over 42 days after undergoing the intervention (0 mm represents no dyspnea and 100 mm represents maximum dyspnea; 10 mm represents minimum clinically significant difference). Mean difference was analyzed using a mixed-effects linear regression model adjusted for minimization variables. Dyspnea improved in both groups, with no significant difference in the first 42 days with a mean VAS dyspnea score of 24.7 in the IPC group (95% CI, 19.3-30.1 mm) and 24.4 mm (95% CI, 19.4-29.4 mm) in the talc group, with a difference of 0.16 mm (95% CI, −6.82 to 7.15; P = .96). There was a statistically significant improvement in dyspnea in the IPC group at 6 months, with a mean difference in VAS score between the IPC group and the talc group of −14.0 mm (95% CI, −25.2 to −2.8 mm; P = .01). Length of initial hospitalization was significantly shorter in the IPC group with a median of 0 days (interquartile range [IQR], 0-1 day) and 4 days (IQR, 2-6 days) for the talc group, with a difference of −3.5 days (95% CI, −4.8 to −1.5 days; P < .001). There was no significant difference in quality of life. Twelve patients (22%) in the talc group required further pleural procedures compared with 3 (6%) in the IPC group (odds ratio [OR], 0.21; 95% CI, 0.04-0.86; P = .03). Twenty-one of the 52 patients in the catheter group experienced adverse events vs 7 of 54 in the talc group (OR, 4.70; 95% CI, 1.75-12.60; P = .002). Among patients with malignant pleural effusion and no previous pleurodesis, there was no significant difference between IPCs and talc pleurodesis at relieving patient-reported dyspnea. isrctn.org Identifier: ISRCTN87514420.


Evaluation of donor cardiac function for heart transplantation: experience of a French academic hospital.
The diagnostic and therapeutic approaches for evaluation and management of cardiac function in brain-dead patients vary from country to country. The aim of the present study was to describe the results of the evaluation of brain-dead patients as potential cardiac donors in a French teaching hospital that manages the largest number of brain-dead patients in France. Demographic parameters, the causes of brain death, clinical evolution, hemodynamic parameters, doses of inotropic and/or vasopressive drugs, the results of echocardiographic examination, and several biochemical markers of myocardial cell injury were retrospectively collected. Seventy-one consecutive brain-dead patients admitted to the intensive care unit of the Academic Hospital of Nancy from October 1st, 1998 to September 30, 1999 were analyzed. Twenty-nine patients were considered as potential heart donors: 22 males and 7 females aged 33 +/- 3 years (Mean + SEM). The cause of brain death was head trauma in 17 cases (59%), cerebrovascular disease in 10 cases (34%), and cerebral anoxia related to cardiac arrest in 2 cases (7%). Eighteen hearts (18/29 or 66%) were harvested and transplanted with a favorable outcome at one month in 17 cases. In 11 cases, the heart was not harvested, nine (9/29 or 31%) because of myocardial dysfunction upon subsequent echocardiographic examination and 2 because of the lack of matched recipients. Comparison of these results with those of other groups suggests that hormonal substitution with insulin and triiodothyronine in the presence of myocardial dysfunction could be of potential interest to correct myocardial dysfunction and increase the number of donor hearts.


Designer photonic dynamics by using non-uniform electron temperature distribution for on-demand all-optical switching times.
While free electrons in metals respond to ultrafast excitation with refractive index changes on femtosecond time scales, typical relaxation mechanisms occur over several picoseconds, governed by electron-phonon energy exchange rates. Here, we propose tailoring these intrinsic rates by engineering a non-uniform electron temperature distribution through nanostructuring, thus, introducing an additional electron temperature relaxation channel. We experimentally demonstrate a sub-300 fs switching time due to the wavelength dependence of the induced hot electron distribution in the nanostructure. The speed of switching is determined by the rate of redistribution of the inhomogeneous electron temperature and not just the rate of heat exchange between electrons and phonons. This effect depends on both the spatial overlap between control and signal fields in the metamaterial and hot-electron diffusion effects. Thus, switching rates can be controlled in nanostructured systems by designing geometrical parameters and selecting wavelengths, which determine the control and signal mode distributions.


Interactions of sodium selenite, glutathione, arsenic species, and omega class human glutathione transferase.
Human monomethylarsenate reductase [MMA(V) reductase] and human glutathione S-transferase omega 1-1 (hGSTO1-1) [because MMA(V) reductase and hGSTO1-1 are identical proteins, the authors will utilize the designation "hGSTO1-1"] are identical proteins that catalyze the reduction of arsenate, monomethylarsenate [MMA(V)], and dimethylarsenate [DMA(V)]. Sodium selenite (selenite) inhibited the reduction of each of these substrates by the enzyme in a concentration-dependent manner. The kinetics indicated a noncompetitive inhibition of the MMA(V), DMA(V), or arsenate reducing activity of hGSTO1-1. The inhibition of the MMA(V) reducting activity of hGSTO1-1 by selenite was reversed by 1 mM DL-dithiothreitol (DTT) but not by reduced glutathione (GSH), which is a required substrate for the enzyme. Neither superoxide anion nor hydrogen peroxide was involved in the selenite inhibition of hGSTO1-1. MALDI-TOF and MS/MS analysis demonstrated that five molecules of GSH were bound to one monomer of hGSTO1-1. Four of the five cysteines of the monomer were glutathionylated. Cys-32 in the active center, however, exists mostly in the sulfhydryl form since it was alkylated consistently by iodoacetamide. MALDI-TOF mass spectra analysis of hGSTO1-1 after reaction with GSH and sodium selenite indicated that selenium was integrated into hGSTO1-1 molecules. Three selenium were found to be covalently bonded to the monomer of hGSTO1-1 with three molecules of GSH. It is proposed that the reaction products of the reduction of selenite inhibited the activity of hGSTO1-1 by reacting with disulfides of glutathionylated cysteines to form bis (S-cysteinyl)selenide and S-selanylcysteine and had little or no interaction with the sulfhydryl of Cys-32 in the active site of the enzyme.


Cardiovascular Disease Risk Assessment in Australian Community Pharmacy.
Population screening and monitoring of cardiovascular risk is suboptimal in Australian primary care. The role of community pharmacy has increased considerably, but without any policy framework for development. The aim of this study was to explore the nature of community pharmacy-based screening models in Australia, capacity to increase delivery of pharmacy screening, and barriers and enablers to increasing capacity. An online survey weblink was emailed to pharmacy managers at every quality-accredited pharmacy in Australia by the Quality Pharmacy Care Program. The 122-item survey explored the nature of screening services, pharmacy capacity to deliver services, and barriers and enablers to service delivery in considerable detail. Adaptive questioning was used extensively to reduce the participant burden. Pharmacy location details were requested to facilitate geo-coding and removal of duplicate entries. A descriptive analysis of responses was undertaken. There were 294 valid responses from 4890 emails, a 6% response rate. Most pharmacies (79%) had private counselling areas. Blood pressure assessment was nearly universal (96%), but other common risk factor assessments were offered by a minority. Most did not charge for assessments, and 59% indicated capacity to provide multiple risk factor assessments. Fewer than one in five (19%) reported any formal arrangements with general practice for care coordination. Financial viability was perceived as a key barrier to service expansion, amid concerns of patient willingness to pay. Support from government and non-governmental organisations for their role was seen as necessary. There appears to be a critical mass of pharmacies engaging in evidence-based and professional services. Considerable additional support appears required to optimise performance across the profession.


Are inland wolf-ungulate systems influenced by marine subsidies of Pacific salmon?
Wolves (Canis lupus) in North America are considered obligate predators of ungulates with other food resources playing little role in wolf population dynamics or wolf prey relations. However, spawning Pacific salmon (Oncorhyncus spp.) are common throughout wolf range in northwestern North America and may provide a marine subsidy affecting inland wolf-ungulate food webs far from the coast. We conducted stable-isotope analyses for nitrogen and carbon to evaluate the contribution of salmon to diets of wolves in Denali National Park and Preserve, 1200 river-km from tidewater in interior Alaska, USA. We analyzed bone collagen from 73 wolves equipped with radio collars during 1986-2002 and evaluated estimates of salmon in their diets relative to the availability of salmon and ungulates within their home ranges. We compared wolf densities and ungulate:wolf ratios among regions with differing salmon and ungulate availability to assess subsidizing effects of salmon on these wolf-ungulate systems. Wolves in the northwestern flats of the study area had access to spawning salmon but low ungulate availability and consumed more salmon (17% +/- 7% [mean +/- SD]) than in upland regions, where ungulates were sixfold more abundant and wolves did or did not have salmon spawning areas within their home ranges (8% +/- 6% and 3% +/- 3%, respectively). Wolves were only 17% less abundant on the northwestern flats compared to the remainder of the study area, even though ungulate densities were 78% lower. We estimated that biomass from fall runs of chum (O. keta) and coho (O. kisutch) salmon on the northwestern flats was comparable to the ungulate biomass there, and the contribution of salmon to wolf diets was similar to estimates reported for coastal wolves in southeast Alaska. Given the ubiquitous consumption of salmon by wolves on the northwestern flats and the abundance of salmon there, we conclude that wolf numbers in this region were enhanced by the allochthonous subsidy provided by salmon and discuss implications for wolf-ungulate relations.


Consumption experience, choice experience and the endowment effect.
We report experiments investigating how experience influences the endowment effect. Our experiments feature endowments which are bundles of unfamiliar consumption goods. We examine how a subject's willingness to swap items from their endowment is influenced by prior experiences of tasting the goods in question and by prior experiences of choosing between them. We do not find a statistically significant endowment effect in our baseline treatment and, because of this, we are unable to test for an effect of consumption experience. We do find an endowment effect when the endowment is acquired in two instalments and, in this setting, we find some evidence that choice experience increases trading. In a follow-up experiment, we find evidence that the absence of an endowment effect in our baseline treatment is due to subjects being more willing to swap when they do not have to give up the last unit of their endowment.


Comparative effects of castration and dehorning in series or concurrent castration and dehorning procedures on stress responses and production in Holstein calves.
The study objective was to compare serum cortisol as an acute stress measure, chute exit velocity as a behavioral measure, and ADG as an indicator of performance and well-being after castration, dehorning, or concurrent castration/dehorning of calves when performed in parallel and in series. Intact male Holstein calves, 3 to 4 mo, underwent sham handling before 2 procedures performed in series separated by 2 to 3 wk. In Period 1, calves were either dehorned by amputation, surgically castrated, concurrently castrated/dehorned, or served as nonsurgical controls (n = 10/treatment). In Period 2, calves that had been dehorned, castrated, or castrated/dehorned were then castrated, dehorned, or served as nonsurgical controls, respectively. Indicators of distress were measured after all procedures; ADG was assessed for 7 d after each procedure and over the 2 to 3 wk interim. Period 1 cortisol concentrations in dehorned calves were less than in castrated and castrated/dehorned calves at 120 min and from 50 to 240 min, respectively (P < 0.02). There was marginal evidence that cortisol concentrations were greater in castrated/dehorned than castrated calves at 60 min (P = 0.06). Period 2 cortisol concentrations were less in dehorned than castrated calves at 120 min (P = 0.005) but were greater from 360 to 480 min (P < 0.002). The Period 2 cortisol profile of control calves did not differ from the baseline obtained during sham handling, despite the intervening castration/dehorning in Period 1, suggesting that memory did not affect cortisol. The cortisol profile of castrated calves did not differ between periods except at 720 min, when Period 1 concentrations were greater than Period 2 (P = 0.02). Cortisol concentrations of calves dehorned in Period 2 were greater than those dehorned in Period 1 at 20 and 240 to 480 min (P < 0.05). In both periods, castrated calves exited the chute slower than dehorned calves (P < 0.05). The ADG did not differ between surgically treated calves in Period 1; in the interim, the ADG of castrated calves was greater than that of castrated/dehorned calves (difference ± SED, 1.4 ± 0.6 kg/d; P = 0.03), and in Period 2, the ADG of dehorned calves was less than castrated calves (1.8 ± 0.6 kg/d; P = 0.005). Our study supports both the common practice of concurrent castration/dehorning and the sequence of dehorning and castration. Delayed dehorning (vs. delayed castration) appeared to be more acutely stressful and more detrimental to ADG.


Meditation programs for psychological stress and well-being: a systematic review and meta-analysis.
Many people meditate to reduce psychological stress and stress-related health problems. To counsel people appropriately, clinicians need to know what the evidence says about the health benefits of meditation. To determine the efficacy of meditation programs in improving stress-related outcomes (anxiety, depression, stress/distress, positive mood, mental health-related quality of life, attention, substance use, eating habits, sleep, pain, and weight) in diverse adult clinical populations. We identified randomized clinical trials with active controls for placebo effects through November 2012 from MEDLINE, PsycINFO, EMBASE, PsycArticles, Scopus, CINAHL, AMED, the Cochrane Library, and hand searches. Two independent reviewers screened citations and extracted data. We graded the strength of evidence using 4 domains (risk of bias, precision, directness, and consistency) and determined the magnitude and direction of effect by calculating the relative difference between groups in change from baseline. When possible, we conducted meta-analyses using standardized mean differences to obtain aggregate estimates of effect size with 95% confidence intervals. After reviewing 18 753 citations, we included 47 trials with 3515 participants. Mindfulness meditation programs had moderate evidence of improved anxiety (effect size, 0.38 [95% CI, 0.12-0.64] at 8 weeks and 0.22 [0.02-0.43] at 3-6 months), depression (0.30 [0.00-0.59] at 8 weeks and 0.23 [0.05-0.42] at 3-6 months), and pain (0.33 [0.03- 0.62]) and low evidence of improved stress/distress and mental health-related quality of life. We found low evidence of no effect or insufficient evidence of any effect of meditation programs on positive mood, attention, substance use, eating habits, sleep, and weight. We found no evidence that meditation programs were better than any active treatment (ie, drugs, exercise, and other behavioral therapies). Clinicians should be aware that meditation programs can result in small to moderate reductions of multiple negative dimensions of psychological stress. Thus, clinicians should be prepared to talk with their patients about the role that a meditation program could have in addressing psychological stress. Stronger study designs are needed to determine the effects of meditation programs in improving the positive dimensions of mental health and stress-related behavior.


A cluster of negative charges at the amino terminal tail of CFTR regulates ATP-dependent channel gating.
1. The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is activated by protein kinase A (PKA) phosphorylation of its R domain and by ATP binding at its nucleotide-binding domains (NBDs). Here we investigated the functional role of a cluster of acidic residues in the amino terminal tail (N-tail) that also modulate CFTR channel gating by an unknown mechanism. 2. A disease-associated mutant that lacks one of these acidic residues (D58N CFTR) exhibited lower macroscopic currents in Xenopus oocytes and faster deactivation following washout of a cAMP -activating cocktail than wild-type CFTR. 3. In excised membrane patches D58N CFTR exhibited a two-fold reduction in single channel open probability due primarily to shortened open channel bursts. 4. Replacing this and two nearby acidic residues with alanines (D47A, E54A, D58A) also reduced channel activity, but had negligible effects on bulk PKA phosphorylation or on the ATP dependence of channel activation. 5. Conversely, the N-tail triple mutant exhibited a markedly inhibited response to AMP-PNP, a poorly hydrolysable ATP analogue that can nearly lock open the wild-type channel. The N-tail mutant had both a slower response to AMP-PNP (activation half-time of 140 +/- 20 s vs. 21 +/- 4 s for wild type) and a lower steady-state open probability following AMP-PNP addition (0.68 +/- 0.08 vs. 0.92 +/- 0.03 for wild type). 6. Introducing the N-tail mutations into K1250A CFTR, an NBD2 hydrolysis mutant that normally exhibits very long open channel bursts, destabilized the activity of this mutant as evidenced by decreased macroscopic currents and shortened open channel bursts. 7. We propose that this cluster of acidic residues modulates the stability of CFTR channel openings at a step that is downstream of ATP binding and upstream of ATP hydrolysis, probably at NBD2.


Heterogeneous muscle gene expression patterns in patients with massive rotator cuff tears.
Detrimental changes in the composition and function of rotator cuff (RC) muscles are hallmarks of RC disease progression. Previous studies have demonstrated both atrophic and degenerative muscle loss in advanced RC disease. However, the relationship between gene expression and RC muscle pathology remains poorly defined, in large part due to a lack of studies correlating gene expression to tissue composition. Therefore, the purpose of this study was to determine how tissue composition relates to gene expression in muscle biopsies from patients undergoing reverse shoulder arthroplasty (RSA). Gene expression related to myogenesis, atrophy and cell death, adipogenesis and metabolism, inflammation, and fibrosis was measured in 40 RC muscle biopsies, including 31 biopsies from reverse shoulder arthroplasty (RSA) cases that had available histology data and 9 control biopsies from patients with intact RC tendons. After normalization to reference genes, linear regression was used to identify relationships between gene expression and tissue composition. Hierarchical clustering and principal component analysis (PCA) identified unique clusters, and fold-change analysis was used to determine significant differences in expression between clusters. We found that gene expression profiles were largely dependent on muscle presence, with muscle fraction being the only histological parameter that was significantly correlated to gene expression by linear regression. Similarly, samples with histologically-confirmed muscle distinctly segregated from samples without muscle. However, two sub-groups within the muscle-containing RSA biopsies suggest distinct phases of disease, with one group expressing markers of both atrophy and regeneration, and another group not significantly different from either control biopsies or biopsies lacking muscle. In conclusion, this study provides context for the interpretation of gene expression in heterogeneous and degenerating muscle, and provides further evidence for distinct stages of RC disease in humans.


Response of cerebral arteries to sympathetic stimulation during acute hypertension.
Our goal was to determine whether sympathetic stimulation during acute hypertension constricts large cerebral arteries and attenuates increases in cerebral microvascular pressure. We measured cerebral blood flow with microspheres and pressure in small pial arteries with a servonull device in anesthetized cats. During moderate hypertension, sympathetic stimulation had little effect on resistance of large or small cerebral vessels. During severe hypertension, sympathetic stimulation prevented passive decreases in resistance of large cerebral arteries and allowed pronounced constriction of small vessels. During hypertension, there was a large increase in pressure in small pial arteries. Although sympathetic stimulation prevented decreases in resistance of large arteries during severe hypertension, it did not attenuate increases in pressure in pial arteries approximately 200 micron in diameter and only modestly attenuated increases in pressure in pial arteries approximately 100 micron in diameter. These findings indicate that sympathetic stimulation has important effects on resistance of both large and small cerebral vessels during severe hypertension. Thus, although stimulation produces dilation of small cerebral vessels during normotension, sympathetic stimulation allowed constriction of small vessels during severe hypertension. These results also indicate that sympathetic stimulation does not prevent increases in pressure in small pial arteries. Thus, protection of the blood-brain barrier by stimulation of sympathetic nerves during hypertension is not the result of attenuation of increases in pial artery pressure.


Magnetic iron oxide nanoparticles for the collection and direct measurement of adsorbed alpha-emitting radionuclides from environmental waters by liquid scintillation analysis.
Radioactive contamination, be it from accidental or intentional release, can create an urgent need to assess water and food supplies and the environment, and monitor human health. In the event of such an emergency, rapid and efficient methods may be needed to assess contamination levels in scores of samples within a short time frame. Internalized exposure to radionuclides that decay by alpha (α) emission can be especially hazardous, given the strongly ionizing nature of the α particle. Unfortunately, the determination of α-emitting radionuclides using traditional radioanalytical methods is typically labor and resource intensive and time consuming. In an effort to devise methods that are fast, require little labor and laboratory expendables, and minimize the use of toxic or corrosive reagents, researchers at PNNL have evaluated superparamagnetic nanoparticles as extracting agents for α-emitting radionuclides from chemically unmodified and acidified (pH 2) aqueous systems. It is demonstrated that bare magnetite nanoparticles exhibit strong affinity for two representative α-emitting radionuclides (241Am and 210Po) from two representative aqueous matrices (river and ground water). Furthermore, use of the superparamagnetic properties of these nanomaterials to concentrate the analyte-bearing solids from the bulk aqueous solution has been demonstrated. The nanoparticle concentrate can be either directly dispensed into a scintillation cocktail, or first dissolved and then added to a scintillation cocktail as a solution for an α-emission assay by liquid scintillation analysis. Despite the severe quenching caused by the metal oxide suspensions in the cocktail, the authors have demonstrated that modern liquid scintillation analyzers can report accurate α activity count rates; the upper limits of nanoparticle suspension concentrations in a cocktail are reported for cases wherein normal instrument count mode and a quench correction protocol are used. Discussions are provided on the presented sample processing and analysis method, the improvement (lowering) of minimum detectable activity concentrations using the nanoparticle-based assay method, and the quenching effects of nanoparticle suspensions in a scintillation cocktail.