DNMT3L stimulates the DNA methylation activity of Dnmt3a and Dnmt3b through a direct interaction.
In mammals, the resetting of DNA methylation patterns in early embryos and germ cells is crucial for development. Two DNA methyltransferases, Dnmt3a and Dnmt3b, are responsible for the creation of DNA methylation patterns. Dnmt3L, a member of the Dnmt3 family, has been reported to be necessary for maternal methylation imprinting, possibly by interacting with Dnmt3a and/or Dnmt3b (Hata, K., Okano, M., Lei, H., and Li, E. (2002) Development 129, 1983-1993). In the present study, the effect of DNMT3L, a human homologue of Dnmt3L, on the DNA methylation activity of mouse Dnmt3a and Dnmt3b was examined in vitro. DNMT3L enhanced the DNA methylation activity of Dnmt3a and Dnmt3b about 1.5-3-fold in a dose-dependent manner but did not enhance the DNA methylation activity of Dnmt1. Although the extents of stimulation were different, a stimulatory effect on the DNA methylation activity was observed for all of the substrate DNA sequences examined, such as those of the maternally methylated SNRPN and Lit-1 imprinting genes, the paternally methylated H19 imprinting gene, the CpG island of the myoD gene, the 5 S ribosomal RNA gene, an artificial 28-bp DNA, poly(dG-dC)-poly(dG-dC), and poly(dI-dC)-poly(dI-dC). DNMT3L could not bind to DNA but could bind to Dnmt3a and Dnmt3b, indicating that the stimulatory effect of DNMT3L on the DNA methylation activity may not be due to the guiding of Dnmt3a and Dnmt3b to the targeting DNA sequence but may comprise a direct effect on their catalytic activity. The carboxyl-terminal half of DNMT3L was found to be responsible for the enhancement of the enzyme activity.


Noncanonical E2 variant-independent function of UBC13 in promoting checkpoint protein assembly.
The E2 ubiquitin-conjugating enzyme UBC13 plays pivotal roles in diverse biological processes. Recent studies have elucidated that UBC13, in concert with the E3 ubiquitin ligase RNF8, propagates the DNA damage signal via a ubiquitylation-dependent signaling pathway. However, mechanistically how UBC13 mediates its role in promoting checkpoint protein assembly and its genetic requirement for E2 variants remain elusive. Here we provide evidence to support the idea that the E3 ubiquitin ligase complex RNF8-UBC13 functions independently of E2 variants and is sufficient in facilitating ubiquitin conjugations and accumulation of DNA damage mediator 53BP1 at DNA breaks. The RNF8 RING domain serves as the molecular platform to anchor UBC13 at the damaged chromatin, where localized ubiquitylation events allow sustained accumulation of checkpoint proteins. Intriguingly, we found that only a group of RING domains derived from E3 ubiquitin ligases, which have been shown to interact with UBC13, enabled UBC13-mediated FK2 and 53BP1 focus formation at DNA breaks. We propose that the RNF8 RING domain selects and loads a subset of UBC13 molecules, distinct from those that exist as heterodimers, onto sites of double-strand breaks, which facilitates the amplification of DNA damage signals.


Characterization of minimum error linear coding with sensory and neural noise.
Robust coding has been proposed as a solution to the problem of minimizing decoding error in the presence of neural noise. Many real-world problems, however, have degradation in the input signal, not just in neural representations. This generalized problem is more relevant to biological sensory coding where internal noise arises from limited neural precision and external noise from distortion of sensory signal such as blurring and phototransduction noise. In this note, we show that the optimal linear encoder for this problem can be decomposed exactly into two serial processes that can be optimized separately. One is Wiener filtering, which optimally compensates for input degradation. The other is robust coding, which best uses the available representational capacity for signal transmission with a noisy population of linear neurons. We also present spectral analysis of the decomposition that characterizes how the reconstruction error is minimized under different input signal spectra, types and amounts of degradation, degrees of neural precision, and neural population sizes.


Molecular basis of androgen resistance in a family with a qualitative abnormality of the androgen receptor and responsive to high-dose androgen therapy.
We have examined the nature of the mutant androgen receptor in a family with a severe defect in virilization associated with a qualitative defect in receptor function. The androgen receptor gene in this family contains two structural alterations: a single nucleotide substitution at position 2444 in exon 5 (adenosine----guanosine) that converts tyrosine 761 to a cysteine residue and a shortened glutamine homopolymeric segment in exon 1 that encodes 12 rather than the usual 20-22 glutamines. A family study was performed using polymerase chain reaction amplification of the glutamine-rich segment, and it was shown that the sister of the proband does not carry the mutant allele. The effects of these two mutations on the function of the androgen receptor were studied by introducing the changes, individually and in combination, into cDNAs encoding the normal human androgen receptor and analyzing the receptor protein produced after transfection of the cDNAs into eukaryotic cells. The presence of a cysteine residue at position 761 causes rapid dissociation of dihydrotestosterone from the receptor protein. Marked thermolability of the transfected receptor protein, however, was demonstrable only upon introduction of an androgen receptor cDNA containing both the partial deletion of the glutamine homopolymeric segment and a cysteine residue at position 761. Likewise, the ability of the receptor to stimulate a reporter gene is strikingly diminished only when both alterations are present, suggesting that the shortened glutamine homopolymeric segment amplifies the impairment of receptor function caused by the tyrosine to cysteine substitution.


Chemopreventive efficacy of Andrographis paniculata on azoxymethane-induced aberrant colon crypt foci in vivo.
Andrographis paniculata is a grass-shaped medicinal herb, traditionally used in Southeast Asia. The aim of this study was to evaluate the chemoprotective effects of A. paniculata on colorectal cancer. A. paniculata ethanol extract was tested on azoxymethane (AOM)-induced aberrant crypt foci (ACF) in vivo and in vitro. A. paniculata treated groups showed a significant reduction in the number of ACF of the treated rats. Microscopically, ACF showed remarkably elongated and stratified cells, and depletion of the submucosal glands of AOM group compared to the treated groups. Histologically, staining showed slightly elevated masses above the surrounding mucosa with oval or slit-like orifices. Immunohistochemically, expression of proliferating cell nuclear antigen (PCNA) and β-catenin protein were down-regulated in the A. paniculata treated groups compared to the AOM group. When colon tissue was homogenized, malondialdehyde (MDA) and nitric oxide (NO) levels were significantly decreased, whereas superoxide dismutase (SOD) activity was increased in the treated groups compared to the AOM group. A. paniculata ethanol extract showed antioxidant and free radical scavenging activity, as elucidated by the measure of oxidative stress markers. Further, the active fractions were assessed against cell lines of CCD841 and HT29 colon cancer cells.


Protective effects of asiatic acid on rotenone- or H2O2-induced injury in SH-SY5Y cells.
Parkinson's disease (PD) is a progressive neurodegenerative disorder with a prevalence of 1-2% in people over the age of 50. Mitochondrial dysfunction occurred in PD patients showing a 15-30% loss of activity in complex I. Asiatic acid (AA), a triterpenoid, is an antioxidant and used for depression treatment, but the effect of AA against PD-like damage has never been reported. In the present study, we investigated the protective effects of AA against H(2)O(2) or rotenone-induced cellular injury and mitochondrial dysfunction in SH-SY5Y cells. Mitochondrial membrane potential (MMP) and the expression of voltage-dependent anion channel (VDAC) were detected with or without AA pretreatment following cellular injury to address the possible mechanisms of AA neuroprotection. The results showed that pre-treatment of AA (0.01-100 nM) protected cells against the toxicity induced by rotenone or H(2)O(2). In addition, MMP dissipation occurred following the exposure of rotenone, which could be prevented by AA treatment. More interestingly, pre-administration of AA inhibited the elevation of VDAC mRNA and protein levels induced by rotenone(100 nM) or H(2)O(2) (300 microM).These data indicate that AA could protect neuronal cells against mitochondrial dysfunctional injury and suggest that AA might be developed as an agent for PD prevention or therapy.


Students Don't Know What They Don't Know: Dental and Oral Health Students' Perspectives on Developing Cultural Competence Regarding Indigenous Peoples.
Indigenous Australians experience poorer oral health than non-Indigenous Australians despite closing-the-gap initiatives. Cultural competence is an important skill in the delivery of oral health care. The need for academic institutions to incorporate Indigenous culture more widely into their curricula to improve educational outcomes for Indigenous peoples and to increase cultural competence for all students has been recognized. The aims of this study were to identify students' perceptions of Indigenous content in current dental and oral health curricula; perceived barriers and supports for developing students' Indigenous cultural competence; and recommended strategies to inform future education in Indigenous culture. Students in the Doctor of Dental Medicine (DMD) and Bachelor of Oral Health (BOH) programs at the University of Sydney participated in semi-structured interviews to explore barriers and supports to students' becoming more competent in Indigenous culture. Thematic analysis was used to synthesize the students' responses. Fifteen students participated in interviews. In analysis of the data, five key themes emerged: defining Indigenous cultural competence; current Indigenous cultural content; barriers to incorporating Indigenous education; future Indigenous curricular content and strategies; and diversity within student cohorts. These findings suggest that increasing Indigenous cultural competence among dental and oral health students requires an informed history of Indigenous Australians, engagement with Indigenous communities, and reflection on these experiences. Additionally, recruitment of Indigenous staff and students in the school will facilitate culturally appropriate ways to redress Indigenous health disparities and increase the overall health of Indigenous peoples.


EAnnot: a genome annotation tool using experimental evidence.
The sequence of any genome becomes most useful for biological experimentation when a complete and accurate gene set is available. Gene prediction programs offer an efficient way to generate an automated gene set. Manual annotation, when performed by experienced annotators, is more accurate and complete than automated annotation. However, it is a laborious and expensive process, and by its nature, introduces a degree of variability not found with automated annotation. EAnnot (Electronic Annotation) is a program originally developed for manually annotating the human genome. It combines the latest bioinformatics tools to extract and analyze a wide range of publicly available data in order to achieve fast and reliable automatic gene prediction and annotation. EAnnot builds gene models based on mRNA, EST, and protein alignments to genomic sequence, attaches supporting evidence to the corresponding genes, identifies pseudogenes, and locates poly(A) sites and signals. Here, we compare manual annotation of human chromosome 6 with annotation performed by EAnnot in order to assess the latter's accuracy. EAnnot can readily be applied to manual annotation of other eukaryotic genomes and can be used to rapidly obtain an automated gene set.


ARF6-dependent regulation of P2Y receptor traffic and function in human platelets.
Adenosine diphosphate (ADP) is a critical regulator of platelet activation, mediating its actions through two G protein-coupled receptors, the P2Y(1) and P2Y(12) purinoceptors. Recently, we demonstrated that P2Y(1) and P2Y(12) purinoceptor activities are rapidly and reversibly modulated in human platelets, revealing that the underlying mechanism requires receptor internalization and subsequent trafficking as an essential part of this process. In this study we investigated the role of the small GTP-binding protein ADP ribosylation factor 6 (ARF6) in the internalization and function of P2Y(1) and P2Y(12) purinoceptors in human platelets. ARF6 has been implicated in the internalization of a number of GPCRs, although its precise molecular mechanism in this process remains unclear. In this study we show that activation of either P2Y(1) or P2Y(12) purinoceptors can stimulate ARF6 activity. Further blockade of ARF6 function either in cell lines or human platelets blocks P2Y purinoceptor internalization. This blockade of receptor internalization attenuates receptor resensitization. Furthermore, we demonstrate that Nm23-H1, a nucleoside diphosphate (NDP) kinase regulated by ARF6 which facilitates dynamin-dependent fission of coated vesicles during endocytosis, is also required for P2Y purinoceptor internalization. These data describe a novel function of ARF6 in the internalization of P2Y purinoceptors and demonstrate the integral importance of this small GTPase upon platelet ADP receptor function.


Outcome of children with low-grade cerebellar astrocytoma: long-term complications and quality of life.
To study the long-term outcome of surgically treated low-grade cerebellar astrocytomas in children. We followed 31 consecutive patients under 16 years of age who were diagnosed between 1980 and 2005 in a single institution. In 21 of 31 survivors (median follow-up time 7.9 years; range 5.6-27.4 years) who agreed to participate, tumor control, neurological and cognitive complications, and their impact on behavioral and emotional adjustment and health-related quality of life (HRQoL) were comprehensively assessed qualitatively and quantitatively. Neurological sequelae were found in 43%. However, age-appropriate ability to perform daily life activities was normal in all patients. Remarkably, cognitive deficits leading to significant school problems occurred in 19% and behavioral and emotional adjustment disturbances in 27%. In comparison with healthy controls, the survivors rated their HRQoL similarly or even higher. Childhood low-grade cerebellar astrocytomas have an excellent cure rate by tumor surgery alone. When compared with other pediatric brain tumors, the risk of neurological, cognitive, emotional, and behavioral complications is relatively small. HRQoL is similar to that of healthy controls.


Graphene-based passively Q-switched dual-wavelength erbium-doped fiber laser.
We demonstrate a compact Q-switched dual-wavelength erbium-doped fiber (EDF) laser based on graphene as a saturable absorber (SA). By optically driven deposition of graphene on a fiber core, the SA is constructed and inserted into a diode-pumped EDF laser cavity. Also benefiting from the strong third-order optical nonlinearity of graphene to suppress the mode competition of EDF, a stable dual-wavelength Q-switching operation has been achieved using a two-reflection peak fiber Bragg grating as the external cavity mirror. The Q-switched EDF laser has a low pump threshold of 6.5 mW at 974 nm and a wide range of pulse-repetition rate from 3.3 to 65.9 kHz. The pulse duration and the pulse energy have been characterized. This is, to the best of our knowledge, the first demonstration of a graphene-based Q-switched laser.


Microbial adhesion on novel yttria-stabilized tetragonal zirconia (Y-TZP) implant surfaces with nitrogen-doped hydrogenated amorphous carbon (a-C:H:N) coatings.
Biomaterial surfaces are at high risk for initial microbial colonization, persistence, and concomitant infection. The rationale of this study was to assess the initial adhesion on novel implant surfaces of Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans upon incubation. The tested samples were 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP) samples with nitrogen-doped hydrogenated amorphous carbon (a-C:H:N) coating (A) and 3Y-TZP samples coated with ceria-stabilized zirconia-based (Ce-TZP) composite and a-C:H:N (B). Uncoated 3Y-TZP samples (C) and bovine enamel slabs (BES) served as controls. Once the surface was characterized, the adherent microorganisms were quantified by estimating the colony-forming units (CFUs). Microbial vitality was assessed by live/dead staining, and microbial-biomaterial surface topography was visualized by scanning electron microscopy (SEM). Overall, A and B presented the lowest CFU values for all microorganisms, while C sheltered significantly less E. faecalis, P. aeruginosa, and C. albicans than BES. Compared to the controls, B demonstrated the lowest vitality values for E. coli (54.12 %) and C. albicans (67.99 %). Interestingly, A (29.24 %) exhibited higher eradication rates for S. aureus than B (13.95 %). Within the limitations of this study, a-C:H:N-coated 3Y-TZP surfaces tended to harbor less initially adherent microorganisms and selectively interfered with their vitality. This could enable further investigation of the new multi-functional zirconia surfaces to confirm their favorable antimicrobial properties in vivo.


Early symbiotic responses induced by Sinorhizobium meliloti iIvC mutants in alfalfa.
A mutation in the ilvC gene of Sinorhizobium meliloti 1021 determines a symbiotically defective phenotype. ilvC mutants obtained from different S. meliloti wild-type strains are able to induce root hair deformation on alfalfa roots and show variable activation of the common nodulation genes nodABC. All of these mutants are noninfective. The presence of extra copies of nodD3-syrM in an IlvC- background does not promote nod expression but allows the detection of low levels of Nod factor production. The sulphation of the Nod factor metabolites, however, is not affected. Furthermore, IlvC- strains induce a specific pattern of starch accumulation on alfalfa roots as well as of early nodulin expression. Hence, the pleiotropic action of the ilvC gene in S. meliloti may reveal novel complexities involved in the symbiotic interaction.


Translating research into practice: transitional care for older adults.
Over the last decade, in order to close the safety and health care quality chasm, there has been a growing imperative to translate evidence-based research into practice. This study examines the major facilitators and barriers of implementing in a large US insurance organization - Aetna Corporation - an evidence-based model of care, the Transitional Care Model, which has been rigorously tested over the past twenty years by a multidisciplinary team at the University of Pennsylvania. Semi-structured interviews of 19 project leaders, case managers, and transitional care nurses were conducted during two phases of translation - start-up and roll out. Qualitative analysis was used to identify more than a dozen key barriers to and facilitators of translation in these two critical phases. Results Six facilitators and seven barriers that are consistent with the literature were identified during and categorized as either start-up or roll-out. The combined results have important practical implications for other, subsequent translational efforts and for assisting providers, policy makers, payers, and other change agents in integrating evidence-based practice with "real world" management.


Prenatal counseling on seat belt use and crash-related medical care.
Describe the prevalence of and characteristics associated with being hurt in a car accident during pregnancy; the resulting medical care needed; and the association between counseling and the medical care needed after a car accident. Secondary data analysis of PRAMS (Pregnancy Risk Assessment Monitoring System) data, a population based survey of women with a recent live birth. Two percent of women were hurt in a car accident during their pregnancy. Only 57% of them had received counseling on seat belt use. Most women (87%) who were hurt in an accident needed medical care. Being counseled on seat belt use was not associated with the level of care needed. Most pregnant women who are hurt in a car accident require medical care or bed rest. Public health action and research is needed to reduce the burden of motor-vehicle related injuries among pregnant women.


Urothelial tumor initiation requires deregulation of multiple signaling pathways: implications in target-based therapies.
Although formation of urothelial carcinoma of the bladder (UCB) requires multiple steps and proceeds along divergent pathways, the underlying genetic and molecular determinants for each step and pathway remain undefined. By developing transgenic mice expressing single or combinatorial genetic alterations in urothelium, we demonstrated here that overcoming oncogene-induced compensatory tumor barriers was critical for urothelial tumor initiation. Constitutively active Ha-ras (Ras*) elicited urothelial hyperplasia that was persistent and did not progress to tumors over a 10 months period. This resistance to tumorigenesis coincided with increased expression of p53 and all pRb family proteins. Expression of a Simian virus 40 T antigen (SV40T), which disables p53 and pRb family proteins, in urothelial cells expressing Ras* triggered early-onset, rapidly-growing and high-grade papillary UCB that strongly resembled the human counterpart (pTaG3). Urothelial cells expressing both Ras* and SV40T had defective G(1)/S checkpoint, elevated Ras-GTPase and hyperactivated AKT-mTOR signaling. Inhibition of the AKT-mTOR pathway with rapamycin significantly reduced the size of high-grade papillary UCB but hyperactivated mitogen-activated protein kinase (MAPK). Inhibition of AKT-mTOR, MAPK and STAT3 altogether resulted in much greater tumor reduction and longer survival than did inhibition of AKT-mTOR pathway alone. Our studies provide the first experimental evidence delineating the combinatorial genetic events required for initiating high-grade papillary UCB, a poorly defined and highly challenging clinical entity. Furthermore, they suggest that targeted therapy using a single agent such as rapamycin may not be highly effective in controlling high-grade UCB and that combination therapy employing inhibitors against multiple targets are more likely to achieve desirable therapeutic outcomes.


Mac-1 deficiency induces respiratory failure by affecting type I alveolar epithelial cells.
As a β2 integrin family member, Mac-1 plays an important role in the inflammatory response. Inflammation and lung injury are closely associated, but the involvement of Mac-1 in the occurrence and development of such pathologies remains poorly understood. We aimed to investigate the relationship between Mac-1 deficiency and respiratory failure in Mac-1 knockout {Mac-1-/-} mice, using C57BL/6J mice as a control. The newborn survival rate of Mac-1-/- mice was calculated, and mouse lung tissue was treated with hematoxylin and eosin and subjected to immunofluorescent staining. Moreover, western blotting and immunohistochemistry were used to detect the expression of molecules specific to type I and type II alveolar epithelial cells, as well as alveolar surfactant proteins secreted by the latter. Survival of Mac-1-/- pups was significantly lower than that of newborn C57BL/6J mice. In a float test, lung tissues from C57BL/6J mice were buoyant, whereas those of Mac-1-/- mice were not. Compared with C57BL/6J mice, expression of proSP-C {specific to type II alveolar epithelial cells} and alveolar surfactant proteins in Mac-1-/- mice was not significantly different, implying that type II cell function was unaltered. However, western blotting revealed expression of T1α, Aqp5, and Snx5 {type I alveolar epithelial cell markers} in Mac-1-/- mice to be significantly decreased {P < 0.05}. In conclusion, Mac-1 may play an important role in respiratory failure. Its absence leads to this condition not by influencing type II alveolar epithelial cells or their secreted surfactant proteins, but rather by reducing type I alveolar cell numbers.


Influence of air-abrasion on zirconia ceramic bonding using an adhesive composite resin.
Air-abrasion as bonding conditioning method for zirconia ceramic might compromise the mechanical strength of zirconia restorations. The purpose of this study was to evaluate the influence of surface conditioning parameters, i.e., air-abrasion with reduced pressure or no air-abrasion and priming with adhesive primers on the long-term resin bond strength to zirconia ceramic. Zirconia ceramic disks were polished with 600 grit abrasive paper. Plexiglas tubes filled with composite resin were bonded with RelyX Unicem luting composite resin to the conditioned zirconia disks. Three surface conditions (unconditioned, air-born particle abrasion at 0.05 or 0.25 MPa) and four priming conditions (no priming, priming with Metal/Zirconia Primer, priming with Alloy Primer, priming with Clearfil Ceramic Primer) were tested. Sixteen specimens of each combination were bonded. Subgroups of eight bonded samples were stored in water either for 3 days or 150 days with 37,500 thermocycling. Tensile bond strengths (TBSs) were determined with a universal testing machine at a crosshead speed of 2mm/min. Without priming, RelyX Unicem showed durable bond strength to 0.25 MPa airborne-particle abraded ceramic. When combined with 10-methacryloyloxy-decyl dihydrogenphosphate containing primers, air-abrasion resulted in a durable TBS to zirconia ceramic even at a reduced abrasion pressure. However, combined with Metal/Zirconia Primer air-abrasion did not provide a durable TBS to zirconia ceramic. Using a self-adhesive luting resin composite (RelyX Unicem), air-abrasion at 0.25 MPa or the combination of low pressure air-abrasion and priming with MDP-containing primers seems to be useful to achieve durable long-term bonding to zirconia ceramic.


Medieval European medicine and Asian spices.
This article aimed to explain the reasons why Asian spices including pepper, ginger, and cinnamon were considered as special and valuable drugs with curative powers in the Medieval Europe. Among these spices, pepper was most widely and frequently used as medicine according to medieval medical textbooks. We analyzed three main pharmacology books written during the Middle Ages. One of the main reasons that oriental spices were widely used as medicine was due to the particular medieval medical system fundamentally based on the humoral theory invented by Hippocrates and Galen. This theory was modified by Arab physicians and imported to Europe during the Middle Ages. According to this theory, health is determined by the balance of the following four humors which compose the human body: blood, yellow bile, black bile, and phlegm. Each humor has its own qualities such as cold, hot, wet, and dry. Humoral imbalance was one of the main causes of disease, so it was important to have humoral equilibrium. Asian spices with hot and dry qualities were used to balance the cold and wet European diet. The analysis of several major medical textbooks of the Middle Ages proves that most of the oriental spices with hot and dry qualities were employed to cure diverse diseases, particularly those caused by coldness and humidity. However, it should be noted that the oriental spices were considered to be much more valuable and effective as medicines than the local medicinal ingredients, which were not only easily procured but also were relatively cheap. Europeans mystified oriental spices, with the belief that they have marvelous and mysterious healing powers. Such mystification was related to the terrestrial Paradise. They believed that the oriental spices were grown in Paradise which was located in the Far East and were brought to the Earthly world along the four rivers flowing from the Paradise.


Validation of a larval zebrafish locomotor assay for assessing the seizure liability of early-stage development drugs.
Seizure liability is an adverse property of new candidate drugs typically detected only in later stage pre-clinical safety studies. Consequently, pharmaceutical discovery needs small scale (microplate-based), rapid throughput screens to 'front-load' such adverse endpoints in order to reduce associated attrition rates. Of the potential methods available, previously published studies have shown that the quantification of seizure-associated locomotion in the larval zebrafish (Danio rerio) offers high potential for development into such a screen. Here, we present methodology and validation data (on 25 compounds) from a larval zebrafish (Zf) convulsant assay, based on the quantification of high speed locomotion after exposure to a range of test compounds. All assays were undertaken in 7 days post fertilization (dpf), WIK-strain Zf larvae, at 27+/-1 degrees C. The blinded validation test set consisted of 17 positive and 8 negative controls, based on literature evidence for seizure liability. Initially, a Maximum Tolerated Concentration (MTC) assay was undertaken on each compound to identify the maximum concentration not causing general toxicity, sedation or overt neuromuscular effects. Next, the convulsant assay was undertaken on 5 concentrations from the MTC down, plus a dilution water control. Exposed larvae were videotracked for 1 h, using the Viewpoint Videotrack for Zebrafish system, and high speed movements, typically associated with seizure-like locomotor activity, were quantified. According to classification criteria proposed by the European Centre for the Validation of Alternative Methods (ECVAM), the data generated appeared to offer "sufficient" predictivity (72% overall), particularly considering the potential for throughput and likely positioning within a safety pharmacology front-loading screening cascade. Possible reasons for the misclassifications are discussed, and potential improvements to increase sensitivity and specificity outlined. In all, these initial validation data suggest that this assay offers potential as a medium throughput screen aimed at the early drug discovery detection of this complex safety pharmacological endpoint.


New and emerging HDAC inhibitors for cancer treatment.
Epigenetic enzymes are often dysregulated in human tumors through mutation, altered expression, or inappropriate recruitment to certain loci. The identification of these enzymes and their partner proteins has driven the rapid development of small-molecule inhibitors that target the cancer epigenome. Herein, we discuss the influence of aberrantly regulated histone deacetylases (HDACs) in tumorigenesis. We examine HDAC inhibitors (HDACis) targeting class I, II, and IV HDACs that are currently under development for use as anticancer agents following the FDA approval of two HDACis, vorinostat and romidepsin.


Smoking prevalence in neighborhood and hospital controls: implications for hospital-based case-control studies.
It is widely believed that the prevalence of smoking among hospital patients is greater than that of the general population because many conditions for which patients are hospitalized are caused by or associated with smoking, and that this increased prevalence may bias results of case-control studies of tobacco-related diseases. For this reason, many authors have suggested excluding from the control series patients hospitalized for tobacco-related illnesses. The present study investigated potential selection bias for hospital compared to neighborhood controls in studying tobacco-related diseases. The 709 cases from six U.S. cities had tobacco-related cancers or myocardial infarction. They were individually matched to one hospital control and to one neighbor. After excluding patients with tobacco-related diseases, hospital controls were less often current smokers and more often former smokers than neighborhood controls. Among male ever smokers, hospital controls tended to smoke more cigarettes per day than neighborhood controls. Compared with the U.S. population, there was an overrepresentation of smokers in neighborhood controls rather than an under-representation of smokers in hospital controls. Relative risk estimates varied according to type of control. Choosing between hospital and neighborhood controls in case-control studies should be dictated by criteria related to the study hypothesis, participation, or cost. In particular, exclusion of hospital controls with diseases known to be tobacco-related seems to be a successful strategy for reducing selection bias.


The vomeronasal organ of Lemur catta.
The vomeronasal organ (VNO), also known as the Jacobson's organ, is a bilateral chemosensory organ found at the base of the nasal cavity specialized for the detection of higher-molecular weight (non-volatile) chemostimuli. It has been linked to pheromone detection. The VNO has been well studied in nocturnal lemurs and lorises, but poorly studied in diurnal/cathemeral species despite the large repertoire of olfactory behaviors noted in species such as Lemur catta. Here, the VNO and associated structures were studied microanatomically in one adult female and one adult male L. catta. Traditional and immunohistochemical procedures demonstrate the VNO epithelium consists of multiple rows of sensory neurons. Immunoreactivity to Growth-associated protein 43 (GAP43) indicates the VNO is postnatally neurogenic. In volume, the VNO neuroepithelium scales similarly to palatal length compared to nocturnal strepsirrhines. Numerous taste buds present at the oral opening to the nasopalatine duct, with which the VNO communicates, provide an additional (or alternative) explanation for the flehmen behavior that has been observed in this species. The VNO of L. catta is shown to be microanatomically comparable to that of nocturnal strepsirrhines. Like nocturnal strepsirrhines, the VNO of L. catta may be functional in the reception of high-molecular weight secretions.


Gas disks to gas giants: simulating the birth of planetary systems.
The ensemble of now more than 250 discovered planetary systems displays a wide range of masses, orbits and, in multiple systems, dynamical interactions. These represent the end point of a complex sequence of events, wherein an entire protostellar disk converts itself into a small number of planetary bodies. Here, we present self-consistent numerical simulations of this process, which produce results in agreement with some of the key trends observed in the properties of the exoplanets. Analogs to our own solar system do not appear to be common, originating from disks near the boundary between barren and (giant) planet-forming.


The Clinical Services Redesign Program in New South Wales: perceptions of senior health managers.
This study explores the views of senior managers regarding their experience of participating in the Clinical Services Redesign Program (CSRP) in New South Wales and the impact of that Program. Semi-structured interviews were conducted in 2007 with 42 senior managers working in the NSW health system. Managers reported being increasingly oriented towards efficiency, achieving results and using data to support decision-making. The increased focus on managing performance was accompanied by concerns about the narrowness of the indicators being used to manage performance and how these are applied. The value placed by interviewees on the use of 'competition' as a lever for improving services varied. Leadership was repeatedly identified as important for long-term success and sustainability. No one was confident that the CSRP had yet been sufficiently embedded in day to day practice in order for it to keep going on its own. Our findings are generally consistent with the extensive literature on change management, performance management and leadership. Some cultural change has taken place in terms of observed patterns of behaviour but it is unrealistic to think that CSRP can on its own deliver the desired deeper cultural changes in the values and assumptions underpinning the NSW Health system. There is some evidence of dysfunctional aspects of performance management but no call for the focus on performance or redesign to be abandoned.


Morphogenesis and cell fate determination within the adaxial cell equivalence group of the zebrafish myotome.
One of the central questions of developmental biology is how cells of equivalent potential-an equivalence group-come to adopt specific cellular fates. In this study we have used a combination of live imaging, single cell lineage analyses, and perturbation of specific signaling pathways to dissect the specification of the adaxial cells of the zebrafish embryo. We show that the adaxial cells are myogenic precursors that form a cell fate equivalence group of approximately 20 cells that consequently give rise to two distinct sub-types of muscle fibers: the superficial slow muscle fibers (SSFs) and muscle pioneer cells (MPs), distinguished by specific gene expression and cell behaviors. Using a combination of live imaging, retrospective and indicative fate mapping, and genetic studies, we show that MP and SSF precursors segregate at the beginning of segmentation and that they arise from distinct regions along the anterior-posterior (AP) and dorsal-ventral (DV) axes of the adaxial cell compartment. FGF signaling restricts MP cell fate in the anterior-most adaxial cells in each somite, while BMP signaling restricts this fate to the middle of the DV axis. Thus our results reveal that the synergistic actions of HH, FGF, and BMP signaling independently create a three-dimensional (3D) signaling milieu that coordinates cell fate within the adaxial cell equivalence group.


Oxidant-induced modifications in the mucosal transcriptome and circulating metabolome of Atlantic salmon.
Here we report the molecular networks associated with the mucosal and systemic responses to peracetic acid (PAA), a candidate oxidative chemotherapeutic in Atlantic salmon (Salmo salar). Smolts were exposed to different therapeutic doses (0, 0.6 and 2.4 mg/L) of PAA for 5 min, followed by a re-exposure to the same concentrations for 30 min 2 weeks later. PAA-exposed groups have higher external welfare score alterations, especially 2 weeks after the re-exposure. Cases of fin damage and scale loss were prevalent in the PAA-exposed groups. Transcriptomic profiling of mucosal tissues revealed that the skin had 12.5 % more differentially regulated genes (DEGs) than the gills following PAA exposure. The largest cluster of DEGs, both in the skin and gills, were involved in tissue extracellular matrix and metabolism. There were 22 DEGs common to both mucosal tissues, which were represented primarily by genes involved in the biophysical integrity of the mucosal barrier, including cadherin, collagen I α 2 chain, mucin-2 and spondin 1a. The absence of significant clustering in the plasma metabolomes amongst the three treatment groups indicates that PAA treatment did not induce any global metabolomic disturbances. Nonetheless, five metabolites with known functions during oxidative stress were remarkably affected by PAA treatments such as citrulline, histidine, tryptophan, methionine and trans-4-hydroxyproline. Collectively, these results indicate that salmon were able to mount mucosal and systemic adaptive responses to therapeutic doses of PAA and that the molecules identified are potential markers for assessing the health and welfare consequences of oxidant exposure.


Interference between storage and processing in working memory: Feature overwriting, not similarity-based competition.
Eight experiments with the complex span paradigm are presented to investigate why concurrent processing disrupts short-term retention. Increasing the pace of the processing task led to worse recall, supporting the hypothesis that the processing task distracts attention from maintenance operations. Neither phonological nor semantic similarity between memory items and processing-task material impaired memory. In contrast, the degree of phonological overlap between memory items and processing-task material affected recall negatively, supporting feature overwriting as one source of interference in the complex span paradigm. When compared directly, phonological overlap impaired memory, but similarity had a beneficial effect. These findings rule out response competition or confusion as a mechanism of interference between storage and processing.


Prognostic factors and survival in primary malignant astrocytomas of the spinal cord: a population-based analysis from 1973 to 2007.
Observational cross-sectional study. Using data from the population-based cancer registries of the Surveillance, Epidemiology and End Results (SEER) program, we analyzed demographic features, tumor and treatment characteristics, as well as survival rates in patients with primary malignant astrocytomas of the spinal cord (PMASC). PMASC is a rare neoplasm and is considered to carry the same dismal outcome as their cerebral counterparts. Our current knowledge is incomplete, and understanding the epidemiology, diagnosis, and optimal treatment still poses challenges. The SEER data from 1973 to 2007 were reviewed for pathologically confirmed primary anaplastic astrocytomas (AA) and glioblastomas of the spinal cord (C72.0). We compared the clinical features and outcomes of the cohort in uni- and multivariate fashion. Survival was calculated and compared using Kaplan-Meier curves and log-rank analysis. Our search criteria retrieved 135 patients diagnosed with PMASC. The median survival for PMASC was 13 months with 1-, 2-, and 5-year survival rates of 51.8%, 32.2%, and 18.7%. Patient diagnosed with AA had a median survival time of 17 months versus 10 months in patients diagnosed with glioblastomas. Adult patients observed markedly prolonged survival compared with the pediatric group, with a 16-month versus 9-month median survival, respectively. Multivariate analysis revealed age at diagnosis, pediatric and adult age groups, sex, tumor histology, and extent of resection as significant predictors of survival. Interestingly, outcomes did not significantly change throughout the last decades or by receiving radiotherapy. Outcome for patients diagnosed with PMASC remains poor and presents an ongoing challenge for professionals in the field of neurospinal medicine and surgery. In our analyses of AA, adult patients, males, and patients undergoing radical resections were associated with increased survival. However, incidence of these lesions is low; hence, building strong collaborative, interdisciplinary, and multi-institutional study groups is necessary to define the optimal treatment of PMASC.


Comparing antimicrobial exposure based on sales data.
This paper explores the possibilities of making meaningful comparisons of the veterinary use of antimicrobial agents among countries, based on national total sales data. Veterinary antimicrobial sales data on country level and animal census data in both Denmark and the Netherlands were combined with information about estimated average dosages, to make model calculations of the average number of treatment days per average animal per year, at first based on the assumption that the treatment incidence is the same in all species and production types. Secondly, the exposure in respectively animals for meat production and dairy and other cattle (excluding veal and young beef) was estimated, assuming zero use in the dairy and other cattle, and thirdly by assuming respectively 100% oral and 100% parenteral administration. Subsequently, the outcomes of these model calculations were compared with treatment incidences calculated from detailed use data per animal species from the national surveillance programmes in these two countries, to assess their accuracy and relevancy. In Denmark and in the Netherlands, although the computed antimicrobial exposure would seem to be a reasonable estimation of the exposure for all animals as a whole, it differs significantly from the measured exposure for most species. The differences in exposure among animal species were much higher than the overall difference between the two countries. For example, the overall model estimate of 9 treatment days per year for Denmark is a severe overestimation of the true use in poultry (i.e. 3 days), and the overall model estimate of 13 treatment days per year for the Netherlands is a severe underestimation of the true use in veal calves (i.e. 66 days). The conclusion is that simple country comparisons, based on total sales figures, entail the risk of serious misinterpretations, especially if expressed in mg per kg. The use of more precise model calculations for making such comparisons, taking into account differences in dosages and in farm animal demographics, only slightly reduces this risk. Overall model estimates are strongly influenced by animal demographics and a very inaccurate indication of the true differences in exposure, per animal species. To get an appropriate certainty about the true differences in antimicrobial exposure between countries it is an absolute necessity to have reliable information about the use per animal species.


An acute psychiatric episode following transvaginal oocyte retrieval.
Psychological disorders of infertile patients are traditionally thought to be chronic, to advance gradually, and to be long-term problems. We describe a patient in whom an acute psychiatric episode developed immediately after transvaginal ultrasound-guided oocyte retrieval. A 34 year old women without history of psychiatric disturbance or adverse reaction to drugs suffered an acute psychiatric episode immediately after oocyte retrieval. She exhibited tachycardia, tachypnoea, transient hypertension and limb rigidity, as well as alterations to stupor and posture. Her vital signs stabilized and she opened her eyes 6 h later, but she persistently raised her head to the left and stared blankly without response to external stimuli. Nine hours later, she was able to look around but remained unresponsive to stimuli. Aphasia was noted in the next morning and a wishful thinking of having delivered a baby was noted in the afternoon. Memory loss was noted on the third day. The patient was diagnosed as having dissociative amnesia and was discharged after three courses of supportive psychotherapy. Assisted reproductive technology-related acute psychiatric episodes, which may initially mimic brainstem stroke, are rare; however, attention should be paid to high-risk patients, and they should be offered elective psychological counselling.


A 1-year study of brimonidine twice daily in glaucoma and ocular hypertension. A controlled, randomized, multicenter clinical trial. Chronic Brimonidine Study Group.
Brimonidin tartrate is a highly selective alpha 2-agonist. This study investigates the safety and efficacy of 0.2% brimonidine administered twice daily for 1 year in patients with glaucoma or ocular hypertension. The study design was a multicenter, double-masked, randomized, parallel-group, active-controlled comparison clinical trial. Subjects instilled 0.2% brimonidine or 0.5% timolol maleate twice daily for 12 months. Subjects were examined at baseline, week 1, and months 1, 2, 3, 6, 9, and 12. A subset of subjects was examined at week 2. Of 443 subjects enrolled in this study, 374 met the entry criteria; 186 received brimonidine and 188 received timolol. Brimonidine-treated subjects showed an overall mean peak reduction in intraocular pressure (IOP) of 6.5 mm Hg; timolol-treated subjects had a mean peak reduction in IOP of 6.1 mm Hg. Brimonidine lowered mean peak IOP significantly more than timolol at week 2 and month 3 (P < .03); no significant difference was observed between the groups for this variable at other visits throughout the 1-year course of the study. No evidence of tachyphylaxis was seen in either group. Allergy was seen in 9% of subjects treated with brimonidine. Dry mouth was more common in the brimonidine-treated group than in the timolol-treated group (33.0% vs 19.4%), but complaints of burning and stinging were more common in the timolol-treated group (41.9%) than in the brimonidine-treated patients (28.1%). Headache, fatigue, and drowsiness were similar in the 2 groups. In general, the tolerance to medication was acceptable. Brimonidine is safe and effective in lowering IOP in glaucomatous eyes. Brimonidine provides a sustained long-term ocular hypotensive effect, is well tolerated, and has a low rate of allergic response.


Perspectives on assistive technology among older Norwegian adults receiving community health services.
The western world is seeking increased implementation of assistive technology (AT) to meet the challenges of an ageing population. The objective of this study is to explore perspectives on AT use among home-dwelling older adults with or without cognitive impairment. This study combines findings from a cross-sectional study with a questionnaire package (n = 83) and from qualitative individual interviews (n = 7) and is part of a larger study, the Assisted Living Project. Combining methods promotes complementary inquiries into a phenomenon. The participants already use ATs: TVs, social alarms, mobile phones, stove timers, electronic medical dispensers, PCs and tablet computers. They were both optimistic and skeptical of AT, and expressed different perspectives and expressed different perspectives on ATs in relation to usability, privacy and fear of losing personal face-to-face care. This study reveals that older adults' perspectives on AT are multifaceted and complex, and can partly be explained by the interacting factors in the HAAT model: person, technology, environment, and context. Further exploration in relation to older adults with health challenges, as well as ethical perspectives on AT implementation, is required for this group. The Norwegian Research Council, Number 47996, funds the Assisted Living Project (ALP).IMPLICATIONS FOR REHABILITATIONThe study was useful in order to inform the health care services about older adults "perspectives on assistive technology".This study reveals the complexity of understanding perspectives towards and the use of assistive technology among older adults with or without cognitive impairment.This study contributes to the understanding of the interactions between the four components: humans, activities, technology and the context.


S9511: a Southwest Oncology Group phase II study of trimetrexate, 5-fluorouracil, and leucovorin in unresectable or metastatic adenocarcinoma of the stomach.
The primary objective of this trial was to evaluate the response rate for trimetrexate in conjunction with 5-FU and leucovorin (LV) (= TFL) in the treatment of advanced gastric cancer in a phase II, cooperative group setting. Patients with locally advanced, unresectable, or metastatic adenocarcinoma of the stomach received trimetrexate 110 mg/m IV over 60 minutes day 1, followed by 5-FU 500 mg/m IV bolus and LV 200 mg/m IV over 60 minutes day 2, followed by oral LV 15 mg every 6 hours x 7 doses, all weekly for 6 weeks followed by 2 weeks of rest, continued until progression. Characteristics for 37 eligible patients: median age 63 (range: 23-83); male/female: 69% of 31%; performance status 0/1/2 15/20/1. The confirmed response rate was 19%, and median overall survival was 6 months. Two patients died as a result of therapy, 1 because of infection without significant neutropenia, and 1 due to perforation of a responding gastric lesion. Seventy-two percent experienced grades 3 and 4 toxicity, most commonly diarrhea, fatigue, and lymphopenia. This regimen achieves response rates comparable to other 5-FU-based regimens, when used in treatment of incurable gastric cancer. Toxicity appears manageable.


ICP-MS/MS-Based Ionomics: A Validated Methodology to Investigate the Biological Variability of the Human Ionome.
We here describe the development, validation and application of a quantitative methodology for the simultaneous determination of 29 elements in human serum using state-of-the-art inductively coupled plasma triple quadrupole mass spectrometry (ICP-MS/MS). This new methodology offers high-throughput elemental profiling using simple dilution of minimal quantity of serum samples. We report the outcomes of the validation procedure including limits of detection/quantification, linearity of calibration curves, precision, recovery and measurement uncertainty. ICP-MS/MS-based ionomics was used to analyze human serum of 120 older adults. Following a metabolomic data mining approach, the generated ionome profiles were subjected to principal component analysis revealing gender and age-specific differences. The ionome of female individuals was marked by higher levels of calcium, phosphorus, copper and copper to zinc ratio, while iron concentration was lower with respect to male subjects. Age was associated with lower concentrations of zinc. These findings were complemented with additional readouts to interpret micronutrient status including ceruloplasmin, ferritin and inorganic phosphate. Our data supports a gender-specific compartmentalization of the ionome that may reflect different bone remodelling in female individuals. Our ICP-MS/MS methodology enriches the panel of validated "Omics" approaches to study molecular relationships between the exposome and the ionome in relation with nutrition and health.


Presence of Broad-Spectrum Beta-Lactamase-Producing Enterobacteriaceae in Zoo Mammals.
Broad-spectrum beta-lactamase (BSBL)-producing Enterobacteriaceae impose public health threats. With increased popularity of zoos, exotic animals are brought in close proximity of humans, making them important BSBL reservoirs. However, not much is known on the presence of BSBLs in zoos in Western Europe. Fecal carriage of BSBL-producing Enterobacteriaceae was investigated in 38 zoo mammals from two Belgian zoos. Presence of bla-genes was investigated using PCR, followed by whole-genome sequencing and Fourier-transform infrared spectroscopy to cluster acquired resistance encoding genes and clonality of BSBL-producing isolates. Thirty-five putatively ceftiofur-resistant isolates were obtained from 52.6% of the zoo mammals. Most isolates were identified as E. coli (25/35), of which 64.0% showed multidrug resistance (MDR). Most frequently detected bla-genes were CTX-M-1 (17/25) and TEM-1 (4/25). Phylogenetic trees confirmed clustering of almost all E. coli isolates obtained from the same animal species. Clustering of five isolates from an Amur tiger, an Amur leopard, and a spectacled bear was observed in Zoo 1, as well as for five isolates from a spotted hyena and an African lion in Zoo 2. This might indicate clonal expansion of an E. coli strain in both zoos. In conclusion, MDR BSBL-producing bacteria were shown to be present in the fecal microbiota of zoo mammals in two zoos in Belgium. Further research is necessary to investigate if these bacteria pose zoonotic and health risks.


Analysis of spinocerebellar ataxia type 1 (SCA1)-related CAG trinucleotide expansion in Japan.
Spinocerebellar ataxia type 1 (SCA1) is a neurodegenerative disorder caused by expansion of a CAG trinucleotide repeat. We analyzed CAG repeat expansion in 25 families in the northeast of Japan with hereditary ataxia of Menzel type. Twenty of 38 patients in 12 families had expanded allele for SCA1. The number of CAG repeats correlated with the age at onset. Although the relationship between anticipation and the number of CAG repeats in successive generations was not ascertainable, there was a tendency to paternal bias for the accelerated age at onset. Study of the number of CAG repeats in various tissues showed no differences in the repeat length in lymphocytes, muscle, or brain; sperm, however, showed an obvious expansion. This may be a clue to a possible mechanism for the molecular basis of paternal anticipation of the disease. The SCA1 gene was transcribed from both wild and mutated alleles in muscles of affected individuals, but the repeat length was the same for both the muscle cDNA and the lymphocyte genomic DNA. These results suggest that, in the area of Japan where SCA1 is prevalent, 48% of families with spinocerebellar degeneration have SCA1 mutation.


Pyrosequencing the transcriptome of the greenhouse whitefly, Trialeurodes vaporariorum reveals multiple transcripts encoding insecticide targets and detoxifying enzymes.
The whitefly Trialeurodes vaporariorum is an economically important crop pest in temperate regions that has developed resistance to most classes of insecticides. However, the molecular mechanisms underlying resistance have not been characterised and, to date, progress has been hampered by a lack of nucleotide sequence data for this species. Here, we use pyrosequencing on the Roche 454-FLX platform to produce a substantial and annotated EST dataset. This 'unigene set' will form a critical reference point for quantitation of over-expressed messages via digital transcriptomics. Pyrosequencing produced around a million sequencing reads that assembled into 54,748 contigs, with an average length of 965 bp, representing a dramatic expansion of existing cDNA sequences available for T. vaporariorum (only 43 entries in GenBank at the time of this publication). BLAST searching of non-redundant databases returned 20,333 significant matches and those gene families potentially encoding gene products involved in insecticide resistance were manually curated and annotated. These include, enzymes potentially involved in the detoxification of xenobiotics and those encoding the targets of the major chemical classes of insecticides. A total of 57 P450s, 17 GSTs and 27 CCEs were identified along with 30 contigs encoding the target proteins of six different insecticide classes. Here, we have developed new transcriptomic resources for T. vaporariorum. These include a substantial and annotated EST dataset that will serve the community studying this important crop pest and will elucidate further the molecular mechanisms underlying insecticide resistance.


Melanoma Metastases to the Adrenal Gland Are Highly Resistant to Immune Checkpoint Inhibitors.
Adrenal gland metastases (AGMs) are common in advanced-stage melanoma, occurring in up to 50% of patients. The introduction of immune checkpoint inhibitors (ICIs) has markedly altered the outcome of patients with melanoma. However, despite significant successes, anecdotal evidence has suggested that treatment responses in AGMs are significantly lower than in other metastatic sites. We sought to investigate whether having an AGM is associated with altered outcomes and whether ICI responses are dampened in the adrenal glands. We retrospectively compared ICI responses and overall survival (OS) in 68 patients with melanoma who were diagnosed with an AGM and a control group of 100 patients without AGMs at a single institution. Response was determined using RECIST 1.1. OS was calculated from time of ICI initiation, anti-PD-1 initiation, initial melanoma diagnosis, and stage IV disease diagnosis. Tumor-infiltrating immune cells were characterized in 9 resected AGMs using immunohistochemical analysis. Response rates of AGMs were significantly lower compared with other metastatic sites in patients with AGMs (16% vs 22%) and compared with those without AGMs (55%). Patients with AGMs also had significantly lower median OS compared with those without AGMs (3.1 years vs not reached, respectively). We further observed that despite this, AGMs exhibited high levels of tumor-infiltrating immune cells. In this cohort of patients with melanoma, those diagnosed with an AGM had lower ICI response rates and OS. These results suggest that tissue-specific microenvironments of AGMs present unique challenges that may require novel, adrenal gland-directed therapies or surgical resection.


Retrotransposon Proliferation Coincident with the Evolution of Dioecy in Asparagus.
Current phylogenetic sampling reveals that dioecy and an XY sex chromosome pair evolved once, or possibly twice, in the genus Asparagus Although there appear to be some lineage-specific polyploidization events, the base chromosome number of 2n = 2× = 20 is relatively conserved across the Asparagus genus. Regardless, dioecious species tend to have larger genomes than hermaphroditic species. Here, we test whether this genome size expansion in dioecious species is related to a polyploidization and subsequent chromosome fusion, or to retrotransposon proliferation in dioecious species. We first estimate genome sizes, or use published values, for four hermaphrodites and four dioecious species distributed across the phylogeny, and show that dioecious species typically have larger genomes than hermaphroditic species. Utilizing a phylogenomic approach, we find no evidence for ancient polyploidization contributing to increased genome sizes of sampled dioecious species. We do find support for an ancient whole genome duplication (WGD) event predating the diversification of the Asparagus genus. Repetitive DNA content of the four hermaphroditic and four dioecious species was characterized based on randomly sampled whole genome shotgun sequencing, and common elements were annotated. Across our broad phylogenetic sampling, Ty-1 Copia retroelements, in particular, have undergone a marked proliferation in dioecious species. In the absence of a detectable WGD event, retrotransposon proliferation is the most likely explanation for the precipitous increase in genome size in dioecious Asparagus species.


Inhibitory effect of liposomal quercetin on acute hepatitis and hepatic fibrosis induced by concanavalin A.
Immune response plays an important role in the development of hepatic fibrosis. In the present study, we investigated the effects of quercetin on hepatitis and hepatic fibrosis induced by immunological mechanism. In the acute hepatitis model, quercetin (2.5 mg/kg) was injected iv into mice 30 min after concanavalin A (Con A) challenge. Mice were sacrificed 4 or 24 h after Con A injection, and aminotransferase tests and histopathological sections were performed. Treatment with quercetin significantly decreased the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Consistent with this observation, treatment with quercetin markedly attenuated the pathologic changes in the liver. A hepatic fibrosis model was also generated in mice by Con A challenge once a week for 6 consecutive weeks. Mice in the experimental group were treated with daily iv injections of quercetin (0.5 mg/kg). Histopathological analyses revealed that treatment with quercetin markedly decreased collagen deposition, pseudolobuli development, and hepatic stellate cells activation. We also examined the effects of quercetin on the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and transforming growth factor beta (TGF-β) pathways by immunohistochemistry and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). NF-κB and TGF-β production was decreased after treatment with quercetin, indicating that the antifibrotic effect of quercetin is associated with its ability to modulate NF-κB and TGF-β production. These results suggest that quercetin may be an effective therapeutic strategy in the treatment of patients with liver damage and fibrosis.


Large Residual Volume, Not Low Packing Density, Is the Most Influential Risk Factor for Recanalization after Coil Embolization of Cerebral Aneurysms.
Tight coil packing with density of at least 20%-25% is known to be important for preventing recanalization after embolization of cerebral aneurysms. However, large aneurysms sometimes recanalize regardless of the packing density, suggesting that the absolute residual volume which is determined by aneurysm volume and packing density may be more important risk factor for recanalization. To validate this hypothesis, we analyzed the factors affecting the outcomes of treated aneurysms at our institute. We included 355 small and large aneurysms. The following six factors were obtained from every case: aneurysm volume (mL), neck size (mm), packing density (%), residual volume (mL), rupture status at presentation, and stent assistance (with or without stent). The data were then subjected to multivariate logistic regression analysis to identify significant risk factors for recanalization. Recanalization occurred in 61 aneurysms (17.2%). Significant predictors for recanalization were aneurysm volume (odds ratio, 15.3; P < 0.001) and residual volume (odds ratio, 30.9; P < 0.001), but not packing density (odds ratio, 0.98; P = 0.341). These results showed that for each 0.1-mL increase in aneurysm volume and residual volume, the risk of recanalization increased by 1.3 times and 1.4 times, respectively. The most influential risk factor for recanalization after coil embolization was residual volume, not packing density. The larger the aneurysm volume, the greater the packing density has to be to minimize the residual volume and risk of recanalization. Since tight coil packing has already been aimed, further innovation of coil property or embolization technique may be needed. Otherwise, different treatment modality such as flow diverter or parent artery occlusion may have to be considered.


Protection of children restrained in child safety seats in side impact crashes.
The performance of child restraint systems (CRS) in side impact motor vehicle crashes has been under study due to the injury and fatality burden of these events. Although previous research has quantified injury risk or described injured body regions, safety advances require an understanding of injury causation. Therefore, the objective was to delineate injury causation scenarios for CRS-restrained children in side impacts and document probable contact points in the vehicle interior. Two in-depth crash investigation databases, the Crash Injury Research and Engineering Network and the Partners for Child Passenger Safety Study, were queried for rear-seated, CRS-restrained children in side impact crashes who sustained Abbreviated Injury Scale 2+ injury. These cases were reviewed by a multidisciplinary team of physicians and engineers to describe injury patterns, injury causation, and vehicle components that contributed to the injuries. Forty-one occupants (average age, 2.6 years) met the inclusion criteria. Twenty-four were near side to the crash, 7 were far side, and 10 were center seated. The most common injuries were to the skull and brain with an increasing proportion of skull fracture as age increased. Head and spine injuries without evidence of head contact were rare but present. All thoracic injuries were lung contusions and no rib fractures occurred. Near-side head and face contacts points were along the rear vertical plane of the window and the horizontal plane of the window sill. Head and face contact points for center- and far-side occupants were along the edges of the front seat back and front seat head restraint. Head injuries are the target for injury prevention for children in CRS in side impact crashes. Most of these injuries are due to the contact; for near-side occupants, contact with the CRS structure and the door interior, for far- or center-seated occupants, contact with the front seat back. These data are useful in developing both educational and technological interventions to reduce the burden of injury to these children.


The contribution of mutant amino acids to alloantigenicity.
The I-Abm12 mutation has been used extensively to study the relationship between structure and function of murine class II major histocompatibility molecules. I-Abm12 differs from I-Ab by three amino acid replacements in the A beta chain, and the proposed structural model of the I-Abm12 molecule places these three amino acid substitutions along one of the alpha-helices where they may affect both antigen and TCR binding. Two of the substitutions, Ile----Phe67 and Thr----Lys71, are thought to point into the binding site, whereas the third substitution, Arg----Gln70, is thought to point up and hence, be available for binding to the TCR. These predicted orientations are consistent with serologic analysis of the bm12 molecule, which demonstrates that residue 70 is uniquely accessible to mAbs distinguishing I-Ab from I-Abm12. In this study we have determined the influence of each of these amino acid substitutions on the ability of the resulting molecules to stimulate a panel of I-Abm12 (allo) reactive T cell hybridomas. Our experiments indicate that reversion of the amino acid at position 70 from Gln (I-Abm12) to Arg (I-Ab) interferes with allorecognition by 33 of 35 I-Abm12-reactive hybridomas. On the other hand, many hybrids can tolerate amino acid substitutions at positions 67 or 71. Single amino acid substitutions at position 67, 70, or 71 are recognized by only a minority of I-Abm12-specific hybrids and usually the reactivity is greatly diminished. These data are most consistent with the idea that the amino acid at position 70 directly interacts with the TCR during allorecognition. The additional effects of residues 67 and 71 are consistent with a contribution by bound peptide to the allorecognition process.


A Twin Study of Perthes Disease.
Legg-Calvé-Perthes disease (LCPD) is an idiopathic avascular necrosis of the femoral head. Its etiology is poorly understood, although previous studies have implicated low birth weight and possible genetic determinants. The aim of this study was to identify potential birth weight and genetic associations with LCPD. We extracted all twin pairs from the Danish Twin Registry (DTR) in which at least 1 individual had LCPD. The DTR captures every twin pair born alive in Denmark, and those with LCPD were identified by using health record linkage. Probanwise concordance was calculated to describe the likelihood that any given individual had LCPD if their co-twin was also diagnosed. There were 81 twin pairs: 10 monozygotic, 51 dizygotic, and 20 unclassified (unknown zygosity [UZ]). There was no association between birth weight and being the affected co-twin. Four pairs (2 dizygotic and 2 UZ) were concordant for LCPD, which is greater than would be expected assuming no familial aggregation. There were no concordant monozygotic twin pairs. The overall probandwise concordance was 0.09 (95% confidence interval [CI]: 0.01-0.18): 0.00 for the monozygotic, 0.08 (95% CI: 0.00-0.18) for the dizygotic, and 0.18 (95% CI: 0.00-0.40) for the UZ twin pairs. This study found evidence of familial clustering in LCPD but did not show a genetic component. The absolute risk that a co-twin of an affected individual will develop LCPD is low, even in the case of monozygotic twin pairs.


Significance of isolated hepatitis B core antibody in blood donors.
About 25% of blood donors who test positive for antibody to hepatitis B core antigen (anti-HBc) have no other positive hepatitis B serologic results. Because of the potential importance and diagnostic uncertainty of this test result, we studied its significance by assessing the serologic response to hepatitis B vaccine in donors with an isolated anti-HBc pattern. Specimens from 300 blood donors that were positive for anti-HBc by enzyme immunoassay were tested for anti-HBc by radioimmunoassay and for antibody to hepatitis B surface antigen (anti-HBs). A subgroup of 37 were further studied after administration of hepatitis B vaccine and compared with 34 similarly vaccinated age- and sex-matched seronegative controls. Measurements of anti-HBs were made at vaccination and 1, 2, 4, 8, 25, and 30 weeks after initial vaccination. Among 300 donors who tested positive for anti-HBc by enzyme immunoassay, the radioimmunoassay for anti-HBc was negative in 76 (25.3%) and the test for anti-HBs was negative in 104 (34.7%). Significant differences were observed for radioimmunoassay anti-HBc and anti-HBs titers, alanine aminotransferase, and male-female ratios between four distinct serogroups (A through D) defined by the combination (positive/negative) of radioimmunoassay anti-HBc and anti-HBs results. No significant differences between the study and control groups were observed in the magnitude of anti-HBs responses at any of the six postvaccine testing periods. Isolated anti-HBc in US blood donors is usually a false-positive result, regardless of the titer.


Electrochemical bacterial detection using poly(3-aminophenylboronic acid)-based imprinted polymer.
Biosensors can deliver the rapid bacterial detection that is needed in many fields including food safety, clinical diagnostics, biosafety and biosecurity. Whole-cell imprinted polymers have the potential to be applied as recognition elements in biosensors for selective bacterial detection. In this paper, we report on the use of 3-aminophenylboronic acid (3-APBA) for the electrochemical fabrication of a cell-imprinted polymer (CIP). The use of a monomer bearing a boronic acid group, with its ability to specifically interact with cis-diol, allowed the formation of a polymeric network presenting both morphological and chemical recognition abilities. A particularly beneficial feature of the proposed approach is the reversibility of the cis-diol-boronic group complex, which facilitates easy release of the captured bacterial cells and subsequent regeneration of the CIP. Staphylococcus epidermidis was used as the model target bacteria for the CIP and electrochemical impedance spectroscopy (EIS) was explored for the label-free detection of the target bacteria. The modified electrodes showed a linear response over the range of 103-107cfu/mL. A selectivity study also showed that the CIP could discriminate its target from non-target bacteria having similar shape. The CIPs had high affinity and specificity for bacterial detection and provided a switchable interface for easy removal of bacterial cell.


Transcription regulators potentially controlled by HPr kinase/phosphorylase in Gram-negative bacteria.
Phosphorylation and dephosphorylation at Ser-46 in HPr, a phosphocarrier protein of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) is controlled by the bifunctional HPr kinase/phosphorylase (HprK/P). In Gram-positive bacteria, P-Ser-HPr controls (1) sugar uptake via the PTS; (2) catabolite control protein A (CcpA)-mediated carbon catabolite repression, and (3) inducer exclusion. Genome sequencing revealed that HprK/P is absent from Gram-negative enteric bacteria, but present in many other proteobacteria. These organisms also possess (1) HPr, the substrate for HprK/P; (2) enzyme I, which phosphorylates HPr at His-15, and (3) one or several enzymes IIA, which receive the phosphoryl group from P approximately His-HPr. The genes encoding the PTS proteins are often organized in an operon with HPRK. However, most of these organisms miss CcpA and a functional PTS, as enzymes IIB and membrane-integrated enzymes IIC seem to be absent. HprK/P and the rudimentary PTS phosphorylation cascade in Gram-negative bacteria must therefore carry out functions different from those in Gram-positive organisms. The gene organization in many HprK/P-containing Gram-negative bacteria as well as some preliminary experiments suggest that HprK/P might control transcription regulators implicated in cell adhesion and virulence. In alpha-proteobacteria, HPRK is located downstream of genes encoding a two-component system of the EnvZ/OmpR family. In several other proteobacteria, HPRK is organized in an operon together with genes from the RPON region of ESCHERICHIA COLI (RPON encodes a sigma54). We propose that HprK/P might control the phosphorylation state of HPr and EIIAs, which in turn could control the transcription regulators.


Phosphorylation Mechanism of Phosphomevalonate Kinase: Implications for Rational Engineering of Isoprenoid Biosynthetic Pathway Enzymes.
The mevalonate pathway is of important clinical, pharmaceutical, and biotechnological relevance. However, lack of the understanding of the phosphorylation mechanism of the kinases in this pathway has limited rationally engineering the kinases in industry. Here the phosphorylation reaction mechanism of a representative kinase in the mevalonate pathway, phosphomevalonate kinase, was studied by using molecular dynamics and hybrid QM/MM methods. We find that a conserved residue (Ser106) is reorientated to anchor ATP via a stable H-bond interaction. In addition, Ser213 located on the α-helix at the catalytic site is repositioned to further approach the substrate, facilitating the proton transfer during the phosphorylation. Furthermore, we elucidate that Lys101 functions to neutralize the negative charge developed at the β-, γ-bridging oxygen atom of ATP during phosphoryl transfer. We demonstrate that the dissociative catalytic reaction occurs via a direct phosphorylation pathway. This is the first study on the phosphorylation mechanism of a mevalonate pathway kinase. The elucidation of the catalytic mechanism not only sheds light on the common catalytic mechanism of the GHMP kinase superfamily but also provides the structural basis for engineering the mevalonate pathway kinases to further exploit their applications in the production of a wide range of fine chemicals such as biofuels or pharmaceuticals.


Next generation sequencing reveals the expression of a unique miRNA profile in response to a gram-positive bacterial infection.
MicroRNAs (miRNAs) are short, non-coding RNAs, which post-transcriptionally regulate gene expression and are proposed to play a key role in the regulation of innate and adaptive immunity. Here, we report a next generation sequencing (NGS) approach profiling the expression of miRNAs in primary bovine mammary epithelial cells (BMEs) at 1, 2, 4 and 6 hours post-infection with Streptococcus uberis, a causative agent of bovine mastitis. Analysing over 450 million sequencing reads, we found that 20% of the approximately 1,300 currently known bovine miRNAs are expressed in unchallenged BMEs. We also identified the expression of more than 20 potentially novel bovine miRNAs. There is, however, a significant dynamic range in the expression of known miRNAs. The top 10 highly expressed miRNAs account for >80% of all aligned reads, with the remaining miRNAs showing much lower expression. Twenty-one miRNAs were identified as significantly differentially expressed post-infection with S. uberis. Several of these miRNAs have characterised roles in the immune systems of other species. This miRNA response to the Gram-positive S. uberis is markedly different, however, to lipopolysaccharide (LPS) induced miRNA expression. Of 145 miRNAs identified in the literature as being LPS responsive, only 9 were also differentially expressed in response to S. uberis. Computational analysis has also revealed that the predicted target genes of miRNAs, which are down-regulated in BMEs following S. uberis infection, are statistically enriched for roles in innate immunity. This suggests that miRNAs, which potentially act as central regulators of gene expression responses to a Gram-positive bacterial infection, may significantly regulate the sentinel capacity of mammary epithelial cells to mobilise the innate immune system.


Reactions to health-related social control in young adults with type 1 diabetes.
Health-related social control refers to individuals' attempts to influence another's health behavior. We describe social control experienced by 109 adults aged 18-35 with Type 1 diabetes, and examine the influence of different types of social control on behavioral and psychological outcomes. Using a self-administered questionnaire, telephone interview, and chart review, we assessed individuals' social control experiences, behavioral and psychological reactions, psychological adjustment, metabolic control, socio-demographics, and clinical factors at baseline, and psychological adjustment and metabolic control at 6-months follow-up. Most participants (85%) reported experiencing social control. Regression analyses revealed that more frequent negative control predicted less behavior change and more negative cognitive reactions concurrently, and decreases in psychological adjustment over time. More frequent reinforcement/modeling and structural changes predicted more positive emotional reactions, but were not associated with behavior change, psychological adjustment, or metabolic control. Use of direct persuasion was associated with more pretending of behavior change. These results suggest that negative social control attempts by social network members may be counter-productive.


Caregivers' contributions to heart failure self-care: a systematic review.
The purpose of this study was to conduct a systematic review answering the following questions: (a) what specific activities do caregivers (CGs) contribute to patients' self-care in heart failure (HF)?; and (b) how mature (or developed) is the science of the CG contribution to self-care? MEDLINE, EMBASE, Cumulative Index of Nursing and Allied Health Literature (CINAHL), the Cochrane Library and ClinicalTrials.gov were searched using the terms heart failure and caregiv* as well as the keywords 'careers', 'family members' and 'lay persons' for studies published between 1948 and September 2012. Inclusion criteria for studies were: informal CGs of adult HF patients-either as dependent/independent variable in quantitative studies or participant in qualitative studies; English language. Exclusion criteria for studies were: formal CGs; pediatric, adult congenital, or devices or transplant CGs; mixed diagnosis; non-empiric reports or reports publishing duplicate results. Each study was abstracted and confirmed by two authors. After CG activities were identified and theoretically categorized, an analysis across studies was conducted. Forty papers were reviewed from a pool of 283 papers. CGs contribute substantively to HF patients' self-care characterized from concrete (weighing the patient) to interpersonal (providing understanding). Only two studies attempted to quantify the impact of CGs' activities on patients' self-care reporting a positive impact. Our analysis provides evidence for a rapidly developing science that is based largely on observational research. To our knowledge, this is the first systematic review to examine CGs' contributions in depth. Informal caregivers play a major role in HF self-care. Longitudinal research is needed to examine the impact of CGs' contributions on patient self-care outcomes.


The association of maternal socialization in childhood and adolescence with adult offsprings' sympathy/caring.
The purpose of the study was to examine associations between mothers' socialization practices in childhood and adolescence and offsprings' (N = 32, 16 female) sympathy/concern in early adulthood. Mothers reported on their socialization practices and beliefs a total of 6 times using a Q-sort during their offsprings' childhood (between 7-8 and 11-12 years of age) and adolescence (between 13-14 and 17-18 years of age). Adult offsprings' sympathy/caring was assessed 3 times in early adulthood (at ages 19-20 to 23-24 years) and in their mid-20s to 30s (ages 25-26 to 31-32 years). In general, friends' reports of participants' sympathy/concern at ages 25-32 years related positively to mother-reported rational discipline (including inductions) and warmth and support during childhood and adolescence and negatively to mother-reported negative affect during adolescence. Self-reported sympathy/concern during early adulthood was positively related to maternal warmth and support during childhood and almost significantly negatively related to mother-reported negative affect during childhood and adolescence. Most of the relations held when the prior level of self-reported childhood empathy or adolescent sympathy was controlled.


Fractures of the greater tuberosity of the humerus.
Isolated fractures of the greater tuberosity of the humerus can occur in anterior shoulder dislocations or as the result of an impaction injury against the acromion or superior glenoid. Greater tuberosity fractures may be associated with partial-thickness rotator cuff tears and labral tears, which may be the cause of persistent pain after fracture healing. Nondisplaced and minimally displaced fractures are typically treated successfully nonsurgically. Surgical fixation is recommended for fractures with >5 mm of displacement in the general population or >3 mm of displacement in active patients involved in frequent overhead activity. Open surgical repair is performed with suture or screw fixation. Recently, arthroscopic techniques have produced promising results. Careful follow-up and supervised rehabilitation optimize results after both nonsurgical and surgical treatment.


Unraveling genetic predisposition to familial or early onset gastric cancer using germline whole-exome sequencing.
Recognition of individuals with a genetic predisposition to gastric cancer (GC) enables preventive measures. However, the underlying cause of genetic susceptibility to gastric cancer remains largely unexplained. We performed germline whole-exome sequencing on leukocyte DNA of 54 patients from 53 families with genetically unexplained diffuse-type and intestinal-type GC to identify novel GC-predisposing candidate genes. As young age at diagnosis and familial clustering are hallmarks of genetic tumor susceptibility, we selected patients that were diagnosed below the age of 35, patients from families with two cases of GC at or below age 60 and patients from families with three GC cases at or below age 70. All included individuals were tested negative for germline CDH1 mutations before or during the study. Variants that were possibly deleterious according to in silico predictions were filtered using several independent approaches that were based on gene function and gene mutation burden in controls. Despite a rigorous search, no obvious candidate GC predisposition genes were identified. This negative result stresses the importance of future research studies in large, homogeneous cohorts.


Uranium Redistribution Due to Water Table Fluctuations in Sandy Wetland Mesocosms.
To understand better the fate and stability of immobilized uranium (U) in wetland sediments, and how intermittent dry periods affect U stability, we dosed saturated sandy wetland mesocosms planted with Scirpus acutus with low levels of uranyl acetate for 4 months before imposing a short drying and rewetting period. Concentrations of U in mesocosm effluent increased after drying and rewetting, but the cumulative amount of U released following the dry period constituted less than 1% of the total U immobilized in the soil during the 4 months prior. This low level of remobilization suggests, and XANES analyses confirm, that microbial reduction was not the primary means of U immobilization, as the U immobilized in mesocosms was primarily U(VI) rather than U(IV). Drying followed by rewetting caused a redistribution of U downward in the soil profile and to root surfaces. Although the U on roots before drying was primarily associated with minerals, the U that relocated to the roots during drying and rewetting was bound diffusely. Results show that short periods of drought conditions in a sandy wetland, which expose reduced sediments to air, may impact U distribution without causing large releases of soil-bound U to surface waters.


Setting thresholds to varying blood pressure monitoring intervals differentially affects risk estimates associated with white-coat and masked hypertension in the population.
Outcome-driven recommendations about time intervals during which ambulatory blood pressure should be measured to diagnose white-coat or masked hypertension are lacking. We cross-classified 8237 untreated participants (mean age, 50.7 years; 48.4% women) enrolled in 12 population studies, using ≥140/≥90, ≥130/≥80, ≥135/≥85, and ≥120/≥70 mm Hg as hypertension thresholds for conventional, 24-hour, daytime, and nighttime blood pressure. White-coat hypertension was hypertension on conventional measurement with ambulatory normotension, the opposite condition being masked hypertension. Intervals used for classification of participants were daytime, nighttime, and 24 hours, first considered separately, and next combined as 24 hours plus daytime or plus nighttime, or plus both. Depending on time intervals chosen, white-coat and masked hypertension frequencies ranged from 6.3% to 12.5% and from 9.7% to 19.6%, respectively. During 91 046 person-years, 729 participants experienced a cardiovascular event. In multivariable analyses with normotension during all intervals of the day as reference, hazard ratios associated with white-coat hypertension progressively weakened considering daytime only (1.38; P=0.033), nighttime only (1.43; P=0.0074), 24 hours only (1.21; P=0.20), 24 hours plus daytime (1.24; P=0.18), 24 hours plus nighttime (1.15; P=0.39), and 24 hours plus daytime and nighttime (1.16; P=0.41). The hazard ratios comparing masked hypertension with normotension were all significant (P<0.0001), ranging from 1.76 to 2.03. In conclusion, identification of truly low-risk white-coat hypertension requires setting thresholds simultaneously to 24 hours, daytime, and nighttime blood pressure. Although any time interval suffices to diagnose masked hypertension, as proposed in current guidelines, full 24-hour recordings remain standard in clinical practice.


The effect of Gibbs ringing artifacts on measures derived from diffusion MRI.
Diffusion-weighted (DW) magnetic resonance imaging (MRI) is a unique method to investigate microstructural tissue properties noninvasively and is one of the most popular methods for studying the brain white matter in vivo. To obtain reliable statistical inferences with diffusion MRI, however, there are still many challenges, such as acquiring high-quality DW-MRI data (e.g., high SNR and high resolution), careful data preprocessing (e.g., correcting for subject motion and eddy current induced geometric distortions), choosing the appropriate diffusion approach (e.g., diffusion tensor imaging (DTI), diffusion kurtosis imaging (DKI), or diffusion spectrum MRI), and applying a robust analysis strategy (e.g., tractography based or voxel based analysis). Notwithstanding the numerous efforts to optimize many steps in this complex and lengthy diffusion analysis pipeline, to date, a well-known artifact in MRI--i.e., Gibbs ringing (GR)--has largely gone unnoticed or deemed insignificant as a potential confound in quantitative DW-MRI analysis. Considering the recent explosion of diffusion MRI applications in biomedical and clinical applications, a systematic and comprehensive investigation is necessary to understand the influence of GR on the estimation of diffusion measures. In this work, we demonstrate with simulations and experimental DW-MRI data that diffusion estimates are significantly affected by GR artifacts and we show that an off-the-shelf GR correction procedure based on total variation already can alleviate this issue substantially.


In vitro analysis of potency restriction during epiblast differentiation.
During mammalian gastrulation, posterior part of primitive ectoderm differentiates into subsets of mesoderm cells, the most proximal portion of which subsequently gives rise to vascular endothelial cells and hematopoietic cells. To analyze this process in which the potency of multipotent epiblast cells is restricted progressively into hematopoietic cell lineages, we have established a culture system that allows differentiation of epiblast cells as well as embryonic stem cells into hematopoietic cells. Using this culture system, we showed that all parts of epiblast tissues at early streak and late bud stages preserved hematogenic potency, while it was lost from anterior region at early head fold stages. However, this loss of hematogenic potency in the anterior region of the head fold stage embryo was reinduced by addition of activin in the culture. Moreover, in order to detect transitory stages of mesoderm cells that are the direct progeny of multipotent epiblast cells, we developed three monoclonal antibodies that are able to define distinct subsets of mesoderm cells in the gastrulating embryo. These results are discussed from a view of cell-mass based commitment.


Photoredox catalysis enabled alkylation of alkenyl carboxylic acids with N-(acyloxy)phthalimide via dual decarboxylation.
A ruthenium based photoredox catalyst in combination with a substoichiometric amount of 1,4-diazabicyclo[2.2.2]octane (DABCO) efficiently catalyzed dual decarboxylative couplings between alkenyl carboxylic acids and N-(acyloxy)phthalimides derived from aliphatic carboxylic acids, delivering alkylated styrene derivatives in a high regio- and stereo-selective manner under mild reaction conditions. Various types of secondary, tertiary, and quaternary aliphatic carboxylic acids as well as α-amino acids can be used as suitable substrates. Mechanistic analysis suggested that the reaction proceeds through a radical mechanism mediated by a Ru(i)/Ru(ii) catalytic cycle with DABCO acting both as the base and the co-catalyst for single electron transfer.


Comparing determinants of alien bird impacts across two continents: implications for risk assessment and management.
Invasive alien species can have serious adverse impacts on both the environment and the economy. Being able to predict the impacts of an alien species could assist in preventing or reducing these impacts. This study aimed to establish whether there are any life history traits consistently correlated with the impacts of alien birds across two continents, Europe and Australia, as a first step toward identifying life history traits that may have the potential to be adopted as predictors of alien bird impacts. A recently established impact scoring system was used in combination with a literature review to allocate impact scores to alien bird species with self-sustaining populations in Australia. These scores were then tested for correlation with a series of life history traits. The results were compared to data from a previous study in Europe, undertaken using the same methodology, in order to establish whether there are any life history traits consistently correlated with impact across both continents. Habitat generalism was the only life history trait found to be consistently correlated with impact in both Europe and Australia. This trait shows promise as a potential predictor of alien bird impacts. The results support the findings of previous studies in this field, and could be used to inform decisions regarding the prevention and management of future invasions.


Echocardiography and circulatory response to progressive endurance exercise.
Cardiac ultrasound techniques have provided an abundance of empirical information regarding normal circulatory responses to dynamic exercise. These data are consistent with a schema by which alterations in peripheral resistance, effected by arteriolar dilatation, facilitate blood flow to exercising muscle and match these responses to increasing metabolic demand. In this model, cardiac responses are governed by quantity of systemic venous return, matched by increases in heart rate to maintain a constant ventricular filling volume. In a progressive test in the upright position, stroke volume rises early, then plateaus as work intensity rises. The initial increase in stroke volume reflects refilling of the heart from the sequestration of blood in the dependent extremities by gravity that occurs when assuming the upright position. Well documented improvements in both inotropic and lusitropic function during progressive exercise therefore serve to maintain constancy of stroke volume and ventricular filling, respectively, with progressive shortening of systolic and diastolic time periods as work intensity rises. During exercise, then, the circulatory system appears to act like an arterial venous fistula, with peripheral resistance serving as the principal factor facilitating and controlling blood flow. Observations in subjects with altered circulatory dynamics during exercise (patients with cardiac disease, highly trained endurance athletes) can be understood within the context of this physiological model.


Gas sorption and luminescence properties of a terbium(III)-phosphine oxide coordination material with two-dimensional pore topology.
The structure, stability, gas sorption properties and luminescence behaviour of a new lanthanide-phosphine oxide coordination material are reported. The polymer PCM-15 is based on Tb(III) and tris(p-carboxylated) triphenylphosphine oxide and has a 5,5-connected net topology. It exhibits an infinite three-dimensional structure that incorporates an open, two-dimensional pore structure. The material is thermally robust and remains crystalline under high vacuum at 150 °C. When desolvated, the solid has a CO(2) BET surface area of 1187 m(2) g(-1) and shows the highest reported uptake of both O(2) and H(2) at 77 K and 1 bar for a lanthanide-based coordination polymer. Isolated Tb(III) centres in the as-synthesized polymer exhibit moderate photoluminescence. However, upon removal of coordinated OH(2) ligands, the luminescence intensity was found to approximately double; this process was reversible. Thus, the Tb(III) centre was used as a probe to detect directly the desolvation and resolvation of the polymer.


Three representations of the Ising model.
Statistical models that analyse (pairwise) relations between variables encompass assumptions about the underlying mechanism that generated the associations in the observed data. In the present paper we demonstrate that three Ising model representations exist that, although each proposes a distinct theoretical explanation for the observed associations, are mathematically equivalent. This equivalence allows the researcher to interpret the results of one model in three different ways. We illustrate the ramifications of this by discussing concepts that are conceived as problematic in their traditional explanation, yet when interpreted in the context of another explanation make immediate sense.