pmid
stringlengths 8
8
| title
stringlengths 9
273
| text
stringlengths 241
3.26k
|
|---|---|---|
23260079 | Hepatitis E virus genotype 4 outbreak, Italy, 2011. | During 2011, 5 persons in the area of Lazio, Italy were infected with a monophyletic strain of hepatitis E virus that showed high sequence homology with isolates from swine in China. Detection of this genotype in Italy parallels findings in other countries in Europe, signaling the possible spread of strains new to Western countries. |
23260059 | Seroepidemiologic effects of influenza A(H1N1)pdm09 in Australia, New Zealand, and Singapore. | To estimate population attack rates of influenza A(H1N1)pdm2009 in the Southern Hemisphere during June-August 2009, we conducted several serologic studies. We pooled individual-level data from studies using hemagglutination inhibition assays performed in Australia, New Zealand, and Singapore. We determined seropositive proportions (titer ≥40) for each study region by age-group and sex in pre- and postpandemic phases, as defined by jurisdictional notification data. After exclusions, the pooled database consisted of, 4,414 prepandemic assays and 7,715 postpandemic assays. In the prepandemic phase, older age groups showed greater seropositive proportions, with age-standardized, community-based proportions ranging from 3.5% in Singapore to 11.9% in New Zealand. In the postpandemic phase, seropositive proportions ranged from 17.5% in Singapore to 30.8% in New Zealand, with highest proportions seen in school-aged children. Pregnancy and residential care were associated with lower postpandemic seropositivity, whereas Aboriginal and Torres Strait Islander Australians and Pacific Peoples of New Zealand had greater postpandemic seropositivity. |
23260060 | Staphylococcal infections in children, California, USA, 1985-2009. | We conducted a retrospective, observational, population-based study to investigate the effect of staphylococcal infections on the hospitalization of children in California during 1985-2009. Hospitalized children with staphylococcal infections were identified through the California Office of Statewide Health Planning and Development discharge database. Infections were categorized as community onset, community onset health care-associated, or hospital onset. Infection incidence was calculated relative to all children and to those hospitalized in acute-care facilities. A total of 140,265 records were analyzed. Overall incidence increased from 49/100,000 population in 1985 to a peak of 83/100,000 in 2006 and dropped to 73/100,000 in 2009. Staphylococcal infections were associated with longer hospital stays and higher risk for death relative to all-cause hospitalizations of children. The number of methicillin-resistant Staphylococcus aureus infections increased, and the number of methicillin-susceptible S. aureus infections remained unchanged. Children <3 years of age, Blacks, and those without private insurance were at higher risk for hospitalization. |
23260058 | Effects of familial mutations on the monomer structure of Aβ₄₂. | Amyloid beta (Aβ) peptide plays an important role in Alzheimer's disease. A number of mutations in the Aβ sequence lead to familial Alzheimer's disease, congophilic amyloid angiopathy, or hereditary cerebral hemorrhage with amyloid. Using molecular dynamics simulations of ∼200 μs for each system, we characterize and contrast the consequences of four pathogenic mutations (Italian, Dutch, Arctic, and Iowa) for the structural ensemble of the Aβ monomer. The four familial mutations are found to have distinct consequences for the monomer structure. |
23260057 | Mathematical study of the role of Delta/Notch lateral inhibition during primary branching of Drosophila trachea development. | A wide range of cellular developmental processes employ intercellular signaling via the Delta/Notch lateral inhibitory pathway to achieve stable spatial patterning. Recent genetic experiments have shown the importance of Delta/Notch lateral inhibition for regulating the number of tip cells in the tracheal primary branching of Drosophila. To examine the role of Delta/Notch regulation in the tip-cell selection, we analyzed a mathematical model of a simple lateral inhibitory system having input signals. Mathematical and numerical analyses revealed that the lateral inhibition did not amplify the signal difference between neighboring cells over the parameter ranges in which the spatial pattern of tip selection was realized. We also show that the number of tip cells becomes less affected by a fluctuation of the input gradient signal as the lateral inhibition becomes stronger. In addition, we demonstrate that the lateral inhibitory regulation enhances the robustness of the tip-cell selection compared with a system regulated by self-inhibition, an alternative means of inhibitory regulation. These results suggest that the lateral inhibition promotes the robustness of tip-cell selection in the tracheal development of Drosophila. |
23260056 | Single-molecule observation of the induction of k-turn RNA structure on binding L7Ae protein. | The k-turn is a commonly occurring structural motif that introduces a tight kink into duplex RNA. In free solution, it can exist in an extended form, or by folding into the kinked structure. Binding of proteins including the L7Ae family can induce the formation of the kinked geometry, raising the question of whether this occurs by passive selection of the kinked structure, or a more active process in which the protein manipulates the RNA structure. We have devised a single-molecule experiment whereby immobilized L7Ae protein binds Cy3-Cy5-labeled RNA from free solution. We find that all bound RNA is in the kinked geometry, with no evidence for transitions to an extended form at the millisecond timescale of the camera. Furthermore, real-time binding experiments provide no evidence for a more extended intermediate even at the earliest times, at a time resolution of 16 ms. The data support a passive conformational selection model by which the protein selects a fraction of RNA that is already in the kinked conformation, thereby drawing the equilibrium into this form. |
23260054 | Mechanistic insights into reversible photoactivation in proteins of the GFP family. | Light-controlled modification of the fluorescence emission properties of proteins of the GFP family is of crucial importance for many imaging applications including superresolution microscopy. Here, we have studied the reversibly photoswitchable fluorescent protein mIrisGFP using optical spectroscopy. By analyzing the pH dependence of isomerization and protonation equilibria and the isomerization kinetics, we have obtained insight into the coupling of the chromophore to the surrounding protein moiety and a better understanding of the photoswitching mechanism. A different acid-base environment of the chromophore's protonating group in its two isomeric forms, which can be inferred from the x-ray structures of IrisFP, is key to the photoswitching function and ensures that isomerization and protonation are correlated. Amino acids near the chromophore, especially Glu212, rearrange upon isomerization, and Glu212 protonation modulates the chromophore pK(a). In mIrisGFP, the cis chromophore protonates in two steps, with pK(cis) of 5.3 and 6, which is much lower than pK(trans) (>10). Based on these results, we have put forward a mechanistic scheme that explains how the combination of isomeric and acid-base properties of the chromophore in its protein environment can produce negative and positive photoswitching modes. |
23260055 | Limitations of time-resolved fluorescence suggested by molecular simulations: assessing the dynamics of T cell receptor binding loops. | Time-resolved fluorescence anisotropy (TRFA) has a rich history in evaluating protein dynamics. Yet as often employed, TRFA assumes that the motional properties of a covalently tethered fluorescent probe accurately portray the motional properties of the protein backbone at the probe attachment site. In an extensive survey using TRFA to study the dynamics of the binding loops of a αβ T cell receptor, we observed multiple discrepancies between the TRFA data and previously published results that led us to question this assumption. We thus simulated several of the experimentally probed systems using a protocol that permitted accurate determination of probe and protein time correlation functions. We found excellent agreement in the decays of the experimental and simulated correlation functions. However, the motional properties of the probe were poorly correlated with those of the backbone of both the labeled and unlabeled protein. Our results warrant caution in the interpretation of TRFA data and suggest further studies to ascertain the extent to which probe dynamics reflect those of the protein backbone. Meanwhile, the agreement between experiment and computation validates the use of molecular dynamics simulations as an accurate tool for exploring the molecular motion of T cell receptors and their binding loops. |
23260053 | Conformational free-energy landscapes for a peptide in saline environments. | The conformations that proteins adopt in solution are a function of both their primary structure and surrounding aqueous environment. Recent experimental and computational work on small peptides, e.g., polyK, polyE, and polyR, have highlighted an interesting and unusual behavior in the presence of aqueous ions such as ClO₄⁻, Na⁺, and K⁺. Notwithstanding the aforementioned studies, as of this writing, the nature of the driving force induced by the presence of ions and its role on the conformational stability of peptides remains only partially understood. Molecular-dynamics simulations have been performed on the heptapeptide AEAAAEA in NaCl and KCl solutions at concentrations of 0.5, 1.0, and 2.0 M. Metadynamics in conjunction with a three-dimensional model reaction coordinate was used to sample the conformational space of the peptide. All simulations were run for 2 μs. Free-energy landscapes were computed over the model reaction coordinate for the peptide in each saline assay as well as in the absence of ions. Circular dichroism spectra were also calculated from each trajectory. In the presence of Na⁺ and K⁺ ions, no increase in helicity is observed with respect to the conformation in pure water. |
23260052 | The effect of DNA CpG methylation on the dynamic conformation of a nucleosome. | DNA methylation is an important epigenetic mark that is known to induce chromatin condensation and gene silencing. We used a time-domain fluorescence lifetime measurement to quantify the effects of DNA hypermethylation on the conformation and dynamics of a nucleosome. Nucleosomes reconstituted on an unmethylated and a methylated DNA both exhibit dynamic conformations under physiological conditions. The DNA end breathing motion and the H2A-H2B dimer destabilization dominate the dynamic behavior of nucleosomes at low to medium ionic strength. Extensive DNA CpG methylation, surprisingly, does not help to restrain the DNA breathing motion, but facilitates the formation of a more open nucleosome conformation. The presence of the divalent cation, Mg(2+), essential for chromatin compaction, and the methyl donor molecule SAM, required for DNA methyltransferase reaction, facilitate the compaction of both types of nucleosomes. The difference between the unmethylated and the methylated nucleosome persists within a broad range of salt concentrations, but vanishes under high magnesium concentrations. Reduced DNA backbone rigidity due to the presence of methyl groups is believed to contribute to the observed structural and dynamic differences. The observation of this study suggests that DNA methylation alone does not compact chromatin at the nucleosomal level and provides molecular details to understand the regulatory role of DNA methylation in gene expression. |
23260051 | Kinetics of the triplex-duplex transition in DNA. | The kinetics of triplex folding/unfolding is investigated by the single-molecule fluorescence resonance energy transfer (FRET) technique. In neutral pH conditions, the average dwell times in both high-FRET (folded) and low-FRET (unfolded) states are comparable, meaning that the triplex is marginally stable. The dwell-time distributions are qualitatively different: while the dwell-time distribution of the high-FRET state should be fit with at least a double-exponential function, the dwell-time distribution of the low-FRET state can be fit with a single-exponential function. We propose a model where the folding can be trapped in metastable states, which is consistent with the FRET data. Our model also accounts for the fact that the relevant timescales of triplex folding/unfolding are macroscopic. |
23260050 | Role of nonspecific interactions in molecular chaperones through model-based bioinformatics. | Molecular chaperones are large proteins or protein complexes from which many proteins require assistance in order to fold. One unique property of molecular chaperones is the cavity they provide in which proteins fold. The interior surface residues which make up the cavities of molecular chaperone complexes from different organisms has recently been identified, including the well-studied GroEL-GroES chaperonin complex found in Escherichia coli. It was found that the interior of these protein complexes is significantly different than other protein surfaces and that the residues found on the protein surface are able to resist protein adsorption when immobilized on a surface. Yet it remains unknown if these residues passively resist protein binding inside GroEL-GroEs (as demonstrated by experiments that created synthetic mimics of the interior cavity) or if the interior also actively stabilizes protein folding. To answer this question, we have extended entropic models of substrate protein folding inside GroEL-GroES to include interaction energies between substrate proteins and the GroEL-GroES chaperone complex. This model was tested on a set of 528 proteins and the results qualitatively match experimental observations. The interior residues were found to strongly discourage the exposure of any hydrophobic residues, providing an enhanced hydrophobic effect inside the cavity that actively influences protein folding. This work provides both a mechanism for active protein stabilization in GroEL-GroES and a model that matches contemporary understanding of the chaperone protein. |
23260049 | Red blood cell membrane dynamics during malaria parasite egress. | Precisely how malaria parasites exit from infected red blood cells to further spread the disease remains poorly understood. It has been shown recently, however, that these parasites exploit the elasticity of the cell membrane to enable their egress. Based on this work, showing that parasites modify the membrane's spontaneous curvature, initiating pore opening and outward membrane curling, we develop a model of the dynamics of the red blood cell membrane leading to complete parasite egress. As a result of the three-dimensional, axisymmetric nature of the problem, we find that the membrane dynamics involve two modes of elastic-energy release: 1), at short times after pore opening, the free edge of the membrane curls into a toroidal rim attached to a membrane cap of roughly fixed radius; and 2), at longer times, the rim radius is fixed, and lipids in the cap flow into the rim. We compare our model with the experimental data of Abkarian and co-workers and obtain an estimate of the induced spontaneous curvature and the membrane viscosity, which control the timescale of parasite release. Finally, eversion of the membrane cap, which liberates the remaining parasites, is driven by the spontaneous curvature and is found to be associated with a breaking of the axisymmetry of the membrane. |
23260048 | Insights into sphingolipid miscibility: separate observation of sphingomyelin and ceramide N-acyl chain melting. | Ceramide produced from sphingomyelin in the plasma membrane is purported to affect signaling through changes in the membrane's physical properties. Thermal behavior of N-palmitoyl sphingomyelin (PSM) and N-palmitoyl ceramide (PCer) mixtures in excess water has been monitored by ²H NMR spectroscopy and compared to differential scanning calorimetry (DSC) data. The alternate use of either perdeuterated or proton-based N-acyl chain PSM and PCer in our ²H NMR studies has allowed the separate observation of gel-fluid transitions in each lipid in the presence of the other one, and this in turn has provided direct information on the lipids' miscibility over a wide temperature range. The results provide further evidence of the stabilization of the PSM gel state by PCer. Moreover, overlapping NMR and DSC data reveal that the DSC-signals parallel the melting of the major component (PSM) except at intermediate (20 and 30 mol %) fractions of PCer. In such cases, the DSC endotherm reports on the presumably highly cooperative melting of PCer. Up to at least 50 mol % PCer, PSM and PCer mix ideally in the liquid crystalline phase; in the gel phase, PCer becomes incorporated into PSM:PCer membranes with no evidence of pure solid PCer. |
23260047 | Detergent properties influence the stability of the glycophorin A transmembrane helix dimer in lysophosphatidylcholine micelles. | Detergents might affect membrane protein structures by promoting intramolecular interactions that are different from those found in native membrane bilayers, and fine-tuning detergent properties can be crucial for obtaining structural information of intact and functional transmembrane proteins. To systematically investigate the influence of the detergent concentration and acyl-chain length on the stability of a transmembrane protein structure, the stability of the human glycophorin A transmembrane helix dimer has been analyzed in lyso-phosphatidylcholine micelles of different acyl-chain length. While our results indicate that the transmembrane protein is destabilized in detergents with increasing chain-length, the diameter of the hydrophobic micelle core was found to be less crucial. Thus, hydrophobic mismatch appears to be less important in detergent micelles than in lipid bilayers and individual detergent molecules appear to be able to stretch within a micelle to match the hydrophobic thickness of the peptide. However, the stability of the GpA TM helix dimer linearly depends on the aggregation number of the lyso-PC detergents, indicating that not only is the chemistry of the detergent headgroup and acyl-chain region central for classifying a detergent as harsh or mild, but the detergent aggregation number might also be important. |
23260046 | Hippocalcin and KCNQ channels contribute to the kinetics of the slow afterhyperpolarization. | The calcium-activated slow afterhyperpolarization (sAHP) is a potassium conductance implicated in many physiological functions of the brain including memory, aging, and epilepsy. In large part, the sAHP's importance stems from its exceedingly long-lasting time-course, which integrates action potential-induced calcium signals and allows the sAHP to control neuronal excitability and prevent runaway firing. Despite its role in neuronal physiology, the molecular mechanisms that give rise to its unique kinetics are, to our knowledge, still unknown. Recently, we identified KCNQ channels as a candidate potassium channel family that can contribute to the sAHP. Here, we test whether KCNQ channels shape the sAHP rise and decay kinetics in wild-type mice and mice lacking Hippocalcin, the putative sAHP calcium sensor. Application of retigabine to speed KCNQ channel activation accelerated the rise of the CA3 pyramidal neuron sAHP current in both wild-type and Hippocalcin knockout mice, indicating that the gating of KCNQ channels limits the sAHP activation. Interestingly, we found that the decay of the sAHP was prolonged in Hippocalcin knockout mice, and that the decay was sensitive to retigabine modulation, unlike in wild-type mice. Together, our results demonstrate that sAHP activation in CA3 pyramidal neurons is critically dependent on KCNQ channel kinetics whereas the identity of the sAHP calcium sensor determines whether KCNQ channel kinetics also limit the sAHP decay. |
23260045 | Spatial and temporal sensing limits of microtubule polarization in neuronal growth cones by intracellular gradients and forces. | Neuronal growth cones are the most sensitive among eukaryotic cells in responding to directional chemical cues. Although a dynamic microtubule cytoskeleton has been shown to be essential for growth-cone turning, the precise nature of coupling of the spatial cue with microtubule polarization is less understood. Here we present a computational model of microtubule polarization in a turning neuronal growth cone. We explore the limits of directional cues in modifying the spatial polarization of microtubules by testing the role of microtubule dynamics, gradients of regulators, and retrograde forces along filopodia. We analyze the steady state and transition behavior of microtubules on being presented with a directional stimulus. Our model makes novel, to our knowledge, predictions about the minimal angular spread of the chemical signal at the growth cone and the fastest polarization times. A regulatory reaction-diffusion network based on the cyclic phosphorylation-dephosphorylation of a regulator predicts that the receptor-signal magnitude can generate the maximal polarization of microtubules and not feedback loops or amplifications in the network. Using both the phenomenological and network models, we have demonstrated some of the physical limits within which the microtubule polarization system works in turning the neuron. |
23260043 | Wanted: a positive control for anomalous subdiffusion. | Anomalous subdiffusion in cells and model systems is an active area of research. The main questions are whether diffusion is anomalous or normal, and if it is anomalous, its mechanism. The subject is controversial, especially the hypothesis that crowding causes anomalous subdiffusion. Anomalous subdiffusion measurements would be strengthened by an experimental standard, particularly one able to cross-calibrate the different types of measurements. Criteria for a calibration standard are proposed. First, diffusion must be anomalous over the length and timescales of the different measurements. The length-scale is fundamental; the time scale can be adjusted through the viscosity of the medium. Second, the standard must be theoretically well understood, with a known anomalous subdiffusion exponent, ideally readily tunable. Third, the standard must be simple, reproducible, and independently characterizable (by, for example, electron microscopy for nanostructures). Candidate experimental standards are evaluated, including obstructed lipid bilayers; aqueous systems obstructed by nanopillars; a continuum percolation system in which a prescribed fraction of randomly chosen obstacles in a regular array is ablated; single-file diffusion in pores; transient anomalous subdiffusion due to binding of particles in arrays such as transcription factors in randomized DNA arrays; and computer-generated physical trajectories. |
23260044 | Force-induced changes in subnuclear movement and rheology. | Extracellular mechanical forces result in changes in gene expression, but it is unclear how cells are able to permanently adapt to new mechanical environments because chemical signaling pathways are short-lived. We visualize force-induced changes in nuclear rheology to examine short- and long-time genome organization and movements. Punctate labels in the nuclear interior of HeLa, human umbilical vein endothelial, and osteosarcoma (Saos-2) cells allow tracking of nuclear movements in cells under varying levels of shear and compressive force. Under adequate shear stress two distinct regimes develop in cells under mechanical stimulation: an initial event of increased intranuclear movement followed by a regime of intranuclear movements that reflect the dose of applied force. At early times there is a nondirectionally oriented response with a small increase in nuclear translocations. After 30 min, there is a significant increase in nuclear movements, which scales with the amount of shear or compressive stress. The similarities in the nuclear response to shear and compressive stress suggest that the nucleus is a mechanosensitive element within the cell. Thus, applied extracellular forces stimulate intranuclear movements, resulting in repositioning of nuclear bodies and the associated chromatin within the nucleus. |
23260040 | Schmallenberg virus in Culicoides spp. biting midges, the Netherlands, 2011. | To determine which species of Culicoides biting midges carry Schmallenberg virus (SBV), we assayed midges collected in the Netherlands during autumn 2011. SBV RNA was found in C. scoticus, C. obsoletus sensu stricto, and C. chiopterus. The high proportion of infected midges might explain the rapid spread of SBV throughout Europe. |
23260039 | Novel framework for assessing epidemiologic effects of influenza epidemics and pandemics. | The effects of influenza on a population are attributable to the clinical severity of illness and the number of persons infected, which can vary greatly between seasons or pandemics. To create a systematic framework for assessing the public health effects of an emerging pandemic, we reviewed data from past influenza seasons and pandemics to characterize severity and transmissibility (based on ranges of these measures in the United States) and outlined a formal assessment of the potential effects of a novel virus. The assessment was divided into 2 periods. Because early in a pandemic, measurement of severity and transmissibility is uncertain, we used a broad dichotomous scale in the initial assessment to divide the range of historic values. In the refined assessment, as more data became available, we categorized those values more precisely. By organizing and prioritizing data collection, this approach may inform an evidence-based assessment of pandemic effects and guide decision making. |
23260038 | The effect of telephone support on depressive symptoms among HIV-infected pregnant women in Thailand: an embedded mixed methods study. | Depressive symptoms negatively impact the lives of HIV-infected individuals and are correlated with faster progression to AIDS. Our embedded mixed methods study examined and described the effects of telephone support on depressive symptoms in a sample of HIV-infected pregnant Thai women. HIV-infected pregnant Thai women (n = 40) were randomly assigned to either the control or the intervention group. A registered nurse provided telephone support to the intervention group. Depressive symptoms were measured at three points in both groups. In-depth interviews were conducted at Time 2 and Time 3. Results show that depressive symptoms in the intervention group decreased over time. Qualitative results describe how telephone support can work, but also reveal that telephone support did not work for everyone. We recommend that a larger mixed methods study be conducted to examine the effects of telephone support on depressive symptoms among HIV-infected women, including the costs and benefits of such support. |
23260037 | Using heteroaryl-lithium reagents as hydroxycarbonyl anion equivalents in conjugate addition reactions with (S,S)-(+)-pseudoephedrine as chiral auxiliary; enantioselective synthesis of 3-substituted pyrrolidines. | We have developed an efficient protocol for carrying out the stereocontrolled formal conjugate addition of hydroxycarbonyl anion equivalents to α,β-unsaturated carboxylic acid derivatives using (S,S)-(+)-pseudoephedrine as chiral auxiliary, making use of the synthetic equivalence between the heteroaryl moieties and the carboxylate group. This protocol has been applied as key step in the enantioselective synthesis of 3-substituted pyrrolidines in which, after removing the chiral auxiliary, the heteroaryl moiety is converted into a carboxylate group followed by reduction and double nucleophilic displacement. Alternatively, the access to the same type of heterocyclic scaffold but with opposite absolute configuration has also been accomplished by making use of the regio- and diastereoselective conjugate addition of organolithium reagents to α,β,γ,δ-unsaturated amides derived from the same chiral auxiliary followed by chiral auxiliary removal, ozonolysis, and reductive amination/intramolecular nucleophilic displacement sequence. |
23260036 | Folic acid supplementation changes the fate of neural progenitors in mouse embryos of hyperglycemic and diabetic pregnancy. | Folic acid has been shown to decrease the incidence of neural tube defects (NTDs) in normal and hyperglycemic conditions, but the influence of folic acid on the development of central nervous system is not fully understood. Here, we aimed to explore the effects of folic acid, especially high dose of folic acid, on the characteristics of neural progenitors in embryos of hyperglycemic and diabetic mouse. Hyperglycemic and diabetic pregnant mice were given 3 mg/kg or 15 mg/kg folic acid from embryonic day 0.5 (E0.5) and were euthanased on E11.5, E13.5 or E18.5. The incidence of NTDs at E13.5 was counted. The proliferation, apoptosis and differentiation of neural progenitors and neuronal migration were determined using BrdU incorporation assay, TUNEL assay, immunofluorescence, Western blot and real-time reverse transcriptase polymerase chain reaction. Both normal and high doses of folic acid decreased the incidence of NTDs, promoted proliferation and reduced apoptosis of neuroepithelial cells in embryos of hyperglycemic and diabetic mice. Importantly, folic acid, especially at high dose, might affect the premature differentiation of neural progenitors in embryos of hyperglycemic and diabetic pregnancy. This may be attributed to changes of messenger RNA expression levels of some basic-helix-loop-helix transcription factors. In addition, folic acid might be involved in regulating neuronal migration in embryos of hyperglycemic and diabetic pregnancy. These findings suggest that periconceptional supplementation of folic acid, especially at high dose, may be a double-edged sword because it may result in undesirable outcomes affecting both the neuronal and glial differentiation in hyperglycemic and diabetic pregnancy. |
23260035 | Stability of AMH measurement in blood and avoidance of proteolytic changes. | The new Gen II assay for anti-Müllerian hormone (AMH) shows good stability and reliability in serum, but analyses of stability in whole blood are lacking. Testing the effects of storage of whole-blood samples at room temperature revealed significant increases in the measured value of AMH of 31% over 4 days (P<0.001). The effect is temperature dependent, with storage at 4°C showing markedly reduced increments. Further, samples collected into serum tubes with gel separators and centrifuged within 5h (blood cells and serum physically separated within the collection tube) showed reliable stability over a period of more than 5 days. |
23260034 | Infertility in resource-constrained settings: moving towards amelioration. | It is often presumed that infertility is not a problem in resource-poor areas where fertility rates are high. This is challenged by consistent evidence that the consequences of childlessness are very severe in low-income countries, particularly for women. In these settings, childless women are frequently stigmatized, isolated, ostracized, disinherited and neglected by the family and local community. This may result in physical and psychological abuse, polygamy and even suicide. Attitudes among people in high-income countries towards provision of infertility care in low-income countries have mostly been either dismissive or indifferent as it is argued that scarce healthcare resources should be directed towards reducing fertility and restricting population growth. However, recognition of the plight of infertile couples in low-income settings is growing. One of the United Nation's Millennium Development Goals was for universal access to reproductive health care by 2015, and WHO has recommended that infertility be considered a global health problem and stated the need for adaptation of assisted reproductive technology in low-resource countries. This paper challenges the construct that infertility is not a serious problem in resource-constrained settings and argues that there is a need for infertility care, including affordable assisted reproduction treatment, in these settings. It is often presumed that infertility is not a problem in densely populated, resource-poor areas where fertility rates are high. This presumption is challenged by consistent evidence that the consequences of childlessness are very severe in low-income countries, particularly for women. In these settings, childless women are frequently stigmatized, isolated, ostracized, disinherited and neglected by the family and local community. This may result in physical and psychological abuse, polygamy and even suicide. Because many families in low-income countries depend on children for economic survival, childlessness and having fewer children than the number identified as appropriate are social and public health matters, not only medical problems. Attitudes among people in high-income countries towards provision of infertility care in low-income countries have mostly been either dismissive or indifferent as it is argued that scarce healthcare resources and family planning activities should be directed towards reducing fertility and restricting population growth. However, recognition of the plight of infertile couples in low-income settings is growing. One of the United Nation's Millennium Development Goals was for universal access to reproductive health care by 2015, and WHO has recommended that infertility be considered a global health problem and stated the need for adaptation of assisted reproduction technology in low-resource countries. In this paper, we challenge the construct that infertility is not a serious problem in resource-constrained settings and argue that there is a need for infertility care, including affordable assisted reproduction treatment, in these settings. |
23260032 | Exploring the toxicity of a bismuth-asparagine coordination polymer on the early development of zebrafish embryos. | Nanoparticles are widely used in nanomedicine, raising concerns about their toxicity. In this study, the toxicity of bismuth-asparagine coordination polymer spheres (BACP-2) was assessed in zebrafish embryos. Injection of 1-4 cell stage embryos with BACP-2 resulted in smaller head size (particularly smaller eye size), shorter body length, and pericardial edemas. The severity and occurrence of the resulting phenotype were concentration-dependent. The expression of genes such as krox20, orthodenticle homeobox 2 (otx2), and cardiac myosin light chain-2 (cmlc2) indicates that the effects of BACP-2 on the head and heart were related to changes in gene expression patterns. A delay in epiboly was observed, and the expression levels of the no tail (ntl) gene indicated that the delay in epiboly resulted both from the effect of BACP-2 on cell migration during epiboly and from slow growth. These findings indicate that BACP-2 exhibits concentration-dependent developmental toxicity, providing insight into the nanotoxicity of bismuth derivatives, which must be rigorously evaluated with respect to toxicity before their application in nanomedicine. |
23260031 | Sheep-to-human transmission of Orf virus during Eid al-Adha religious practices, France. | Five persons in France were infected with Orf virus after skin wounds were exposed to infected sheep tissues during Eid al-Adha, the Muslim Feast of Sacrifice. Infections were confirmed by electron microscopy, PCR, and sequence analysis. Prevention and control of this underdiagnosed disease can be achieved by educating physicians, slaughterhouse workers, and persons participating in Eid al-Adha. |
23260030 | Abundance, distribution and potential impact of transposable elements in the genome of Mycosphaerella fijiensis. | Mycosphaerella fijiensis is a ascomycete that causes Black Sigatoka in bananas. Recently, the M. fijiensis genome was sequenced. Repetitive sequences are ubiquitous components of fungal genomes. In most genomic analyses, repetitive sequences are associated with transposable elements (TEs). TEs are dispersed repetitive DNA sequences found in a host genome. These elements have the ability to move from one location to another within the genome, and their insertion can cause a wide spectrum of mutations in their hosts. Some of the deleterious effects of TEs may be due to ectopic recombination among TEs of the same family. In addition, some transposons are physically linked to genes and can control their expression. To prevent possible damage caused by the presence of TEs in the genome, some fungi possess TE-silencing mechanisms, such as RIP (Repeat Induced Point mutation). In this study, the abundance, distribution and potential impact of TEs in the genome of M. fijiensis were investigated. A total of 613 LTR-Gypsy and 27 LTR-Copia complete elements of the class I were detected. Among the class II elements, a total of 28 Mariner, five Mutator and one Harbinger complete elements were identified. The results of this study indicate that transposons were and are important ectopic recombination sites. A distribution analysis of a transposable element from each class of the M. fijiensis isolates revealed variable hybridization profiles, indicating the activity of these elements. Several genes encoding proteins involved in important metabolic pathways and with potential correlation to pathogenicity systems were identified upstream and downstream of transposable elements. A comparison of the sequences from different transposon groups suggested the action of the RIP silencing mechanism in the genome of this microorganism. The analysis of TEs in M. fijiensis suggests that TEs play an important role in the evolution of this organism because the activity of these elements, as well as the rearrangements caused by ectopic recombination, can result in deletion, duplication, inversion and translocation. Some of these changes can potentially modify gene structure or expression and, thus, facilitate the emergence of new strains of this pathogen. |
23260029 | Novel polyomavirus associated with brain tumors in free-ranging raccoons, western United States. | Tumors of any type are exceedingly rare in raccoons. High-grade brain tumors, consistently located in the frontal lobes and olfactory tracts, were detected in 10 raccoons during March 2010-May 2012 in California and Oregon, suggesting an emerging, infectious origin. We have identified a candidate etiologic agent, dubbed raccoon polyomavirus, that was present in the tumor tissue of all affected animals but not in tissues from 20 unaffected animals. Southern blot hybridization and rolling circle amplification showed the episomal viral genome in the tumors. The multifunctional nuclear protein large T-antigen was detectable by immunohistochemical analyses in a subset of neoplastic cells. Raccoon polyomavirus may contribute to the development of malignant brain tumors of raccoons. |
23260028 | Improvement of conception rate in postpartum flaxseed supplemented buffalo with Ovsynch+CIDR protocol. | The present study was conducted on lactating Murrah buffalo to assess the effect of crushed flaxseed (a source of omega-3 fatty acids) supplementation (300g/100kg bwt/day for 60 days), over and above the routine feed, on luteolytic signal (PGF2α), luteal function (progesterone) and conception rate. In first experiment, on day 50 post-calving, six non-supplemented buffalo were treated to synchronize time of ovulation using an Ovsynch+Controlled Internal Drug Release (CIDR) protocol followed by intravenous oxytocin treatment (OT; 100IU) on day 15 post-ovulation. Blood samples were collected at 15min interval, 1h before to 4h after OT challenge. Thereafter, the same buffalo were supplemented with flaxseed, treated to synchronize time of ovulation starting on day 35 post-supplementation using the same protocol and subjected to OT treatment and blood sampling on day 15 post-ovulation. The PGF2α response was measured as the venous concentration of 13,14-dihydro-15-keto PGF2α (PGFM). The mean hourly concentration of PGFM subsequent to flaxseed supplemented was less (P<0.05) than in the pre-supplementation period at all the occasions. Flaxseed supplementation did not affect plasma fatty acids and other plasma metabolites except for an increase (P<0.05) in plasma cholesterol and plasma alanine transaminase. In the second experiment, 31 buffalo were randomly assigned to a control (n=16) and flaxseed supplemented (n=15) group. The latter group was supplemented with flaxseed starting from day 15 post-calving. On day 50-post-calving, buffalo of both groups were treated to synchronize time of ovulation among animals as described for the first experiment followed by artificial insemination (AI). Post-AI luteal phase plasma progesterone was greater (P<0.05) in the supplemented group compared to controls. Conception rate on day 63 post-AI was 66.7% in supplemented and 31.2% in controls (P<0.05). The present study indicated the beneficial impact of dietary supplementation of crushed flaxseed on conception rate through attenuation of luteolytic signal and improvement in post-breeding luteal profile. |
23260026 | Approaches to virtual screening and screening library selection. | The ease of access to virtual screening (VS) software in recent years has resulted in a large increase in literature reports. Over 300 publications in the last year report the use of virtual screening techniques to identify new chemical matter or present the development of new virtual screening techniques. The increased use is accompanied by a corresponding increase in misuse and misinterpretation of virtual screening results. This review aims to identify many of the common difficulties associated with virtual screening and allow researchers to better assess the reliability of their virtual screening effort. |
23260025 | Advances in binding free energies calculations: QM/MM-based free energy perturbation method for drug design. | Multiple approaches have been devised and evaluated to computationally estimate binding free energies. Results using a recently developed Quantum Mechanics (QM)/Molecular Mechanics (MM) based Free Energy Perturbation (FEP) method suggest that this method has the potential to provide the most accurate estimation of binding affinities to date. The method treats ligands/inhibitors using QM while using MM for the rest of the system. The method has been applied and validated for a structurally diverse set of fructose 1,6- bisphosphatase (FBPase) inhibitors suggesting that the approach has the potential to be used as an integral part of drug discovery for both lead identification lead optimization, where there is a structure available. In addition, this QM/MM-based FEP method was shown to accurately replicate the anomalous hydration behavior exhibited by simple amines and amides suggesting that the method may also prove useful in predicting physical properties of molecules. While the method is about 5-fold more computationally demanding than conventional FEP, it has the potential to be less demanding on the end user since it avoids development of MM force field parameters for novel ligands and thereby eliminates this time-consuming step that often contributes significantly to the inaccuracy of binding affinity predictions using conventional FEP methods. The QM/MM-based FEP method has been extensively tested with respect to important considerations such as the length of the simulation required to obtain satisfactory convergence in the calculated relative solvation and binding free energies for both small and large structural changes between ligands. Future automation of the method and parallelization of the code is expected to enhance the speed and increase its use for drug design and lead optimization. |
23260024 | Design of glycogen synthase kinase-3 inhibitors: an overview on recent advancements. | Glycogen Synthase Kinase-3 (GSK-3) is a constitutively acting multifunctional serine/threonine kinase, a role of which has been marked in several physiological pathways, making it a potential target for the treatment of many diseases, including Type-II diabetes and Alzheimer's. Design of GSK-3β selective inhibitor was the key challenge which led to the use of rational approaches like structure based methods (molecular docking), and ligand based methods (QSAR, pharmacophore mapping) studies. These methods provide insights into the enzyme-ligand interactions and structure activity relationship of different sets of compounds for the design of promising GSK-3 inhibitors. Molecular dynamic simulation studies have additionally been performed to address key issues like the unique requirement of prime phosphorylation of its substrate at P+4 by GSK-3β. An allosteric site has also been reported, where the binding of the peptide leads to the stabilization of the activation loop, resulting in the enhancement of the catalysis of enzymes. These studies are becoming useful in the design of therapeutically active discriminatory GSK-3 inhibitors. In this article, we present a review of recent efforts and future opportunities for the design of selective GSK-3β inhibitors. |
23260023 | Selectivity, binding affinity, and ionization state of matrix metalloproteinase inhibitors. | This review highlights some recent advances in the design and development of matrix metalloproteinase inhibitors, especially those targeting MMP-2, MMP-9, and MMP-13. Various zinc-binding groups and non-zinc-binding groups are discussed. Interactions between residues in the critical S1' specificity pocket and MMP inhibitors are given special attention. The influence of ionization states of hydroxamates and retrohydroxamates on the docking outcome and the presence of zinc ions in the active site are explored in light of enhancing enrichment factors for docking studies. Details are given to structural factors for the development of more selective and more potent MMP inhibitors. |
23260020 | Genomes to hits in silico - a country path today, a highway tomorrow: a case study of chikungunya. | These are exciting times for bioinformaticians, computational biologists and drug designers with the genome and proteome sequences and related structural databases growing at an accelerated pace. The post-genomic era has triggered high expectations for a rapid and successful treatment of diseases. However, in this biological information rich and functional knowledge poor scenario, the challenges are indeed grand, no less than the assembly of the genome of the whole organism. These include functional annotation of genes, identification of druggable targets, prediction of three-dimensional structures of protein targets from their amino acid sequences, arriving at lead compounds for these targets followed by a transition from bench to bedside. We propose here a "Genome to Hits In Silico" strategy (called Dhanvantari) and illustrate it on Chikungunya virus (CHIKV). "Genome to hits" is a novel pathway incorporating a series of steps such as gene prediction, protein tertiary structure determination, active site identification, hit molecule generation, docking and scoring of hits to arrive at lead compounds. The current state of the art for each of the steps in the pathway is high-lighted and the feasibility of creating an automated genome to hits assembly line is discussed. |
23260022 | Rational approaches towards lead optimization of kinase inhibitors: the issue of specificity. | Kinases are one of the most popular classes of drug targets as they are involved in signal transduction pathways, which are wired through a phosphotransfer cascade and elicit a number of important and essential physiological responses. Kinase specificity has emerged as one of the major issues to be addressed in drug discovery approaches. In most kinases the active site is the ATP binding site and finding suitable hits which maximize the affinity of binding has been traditionally important to obtain the type I inhibitors. While type I inhibitors have effective binding affinity more often than not they encounter side-effects usually associated with specificity. Therefore in recent times it has become indispensable to optimize specificity for developing effective kinase inhibitors. The review presents an overview of kinase drug discovery and the different strategies used to date for the design of kinase leads accounting for their success and failure. A number of strategies exploiting different aspects of kinases like allosteric site, size of the gatekeeper residue, DFG-loop, chemotype selectivity, non-covalent interactions, salt-bridge, solvation, etc. have been explored to circumvent the specificity problem in kinases. The probable hot-spots in kinases having a propensity to bring in specificity have been delineated with special emphasis on the design of type II inhibitors with increased specificity from existing type I using fragment tailoring approach. In this review we illustrate the current strategies by taking p38 MAP kinase as a model and expect that such strategies are general and can be extended to the other members of the kinase family. |
23260021 | Drug design for protein kinases and phosphatases: flexible-receptor docking, binding affinity and specificity, and drug-binding kinetics. | This article reviews some of our experiences on applying computational techniques to aid the design of drugs targeting protein kinases and phosphatases. It is not a comprehensive review. Rather, it focuses on several less explored approaches or ideas that we have experiences on. It reviews some recent improvements on the Poisson-Boltzmann/Surface Area model for calculating binding affinity and discusses ways to perform calculations that are more tolerant to statistical and systematic errors. Several new ways to incorporate protein flexibility in molecular docking and estimating binding affinity are also discussed. Its discussions also go beyond binding affinity to considering drug-binding kinetics, not only on investigating protein-ligand interactions in isolation, but also on accounting for upstream and downstream influences that can occur in cells, through kinetic modeling of cell signaling. This review also describes a quick molecular simulation method for understanding drug-binding kinetics at the molecular level, with the hope of generating guiding principles for designing drugs with the desired kinetic properties. Sources of drug-binding selectivity that appear obvious but often overlooked are also discussed. |
23260019 | HIV-1 associated Topoisomerase IIβ kinase: a potential pharmacological target for viral replication. | Viruses have been found to exhibit protein kinase activity associated with their purified viral particles. HIV-1 virus particles possess a novel 72 kD protein, Topoisomerase II beta kinase (Topo IIβKHIV) activity. The enzyme, isolated and purified from PEGprecipitated HIV-1 particles, is insensitive against a diverse set of known kinase inhibitors. The pyridine derivatives were found to be active against both Topo IIβKHIV activity and HIV-1 replication. For both kinase antagonism and anti-HIV-1 activity the Comparative Molecular Field Analysis (CoMFA) models were proposed. The CoMFA model was also evaluated independently with a set of test molecules for their anti-viral activity. The kinase inhibition and anti-viral activities for these inhibitors, tested in an in vitro kinase agree with the CoMFA model (cross-validated r2 (q2) value of 0.642 with six principal components), lower acceptable results are obtained with anti- HIV-1 activity (cross-validated r2 (q2) value of 0.358 with four principal components) and also correlate with relative solvation free energy calculations. The predictive power of the models was evaluated with 2 test molecules each and tends to lie within 1 log unit. An in cell validation of the model with a representative inhibitor, 2-methoxypyridine shows its ability to inhibit Topo IIβ phosphorylation during acute HIV-1 infection. Close correlation of molecular fields of inhibitory domains of kinase and HIV-1 inhibitors suggests specificity of action of pyridine derivatives in affecting HIV-1 replication through inhibition of Kinase activity. These investigations suggest that Topo IIβKHIV is a potential target for an effective control of HIV-1 replication that would help in developing new anti-retroviral molecules. |
23260016 | Ca(2+) homeostasis and regulation of ER Ca(2+) in mammalian oocytes/eggs. | The activation of the developmental program in mammalian eggs relies on the initiation at the time of fertilization of repeated rises in the intracellular concentration of free calcium ([Ca(2+)](i)), also known as [Ca(2+)](i) oscillations. The ability to mount the full complement of oscillations is only achieved at the end of oocyte maturation, at the metaphase stage of meiosis II (MII). Over the last decades research has focused on addressing the mechanisms by which the sperm initiates the oscillations and identification of the channels that mediate intracellular Ca(2+) release. This review will describe the up-to-date knowledge of other aspects of Ca(2+) homeostasis in mouse oocytes, such as the mechanisms that transport Ca(2+) out of the cytosol into the endoplasmic reticulum (ER), the Ca(2+) store of the oocyte/egg, into other organelles and also those that extrude Ca(2+). Evidence pointing to channels in the plasma membrane that mediate Ca(2+) entry from the extracellular milieu, which is required for the persistence of the oscillations, is also discussed, along with the modifications that these mechanisms undergo during maturation. Lastly, we highlight areas where additional research is needed to obtain a better understating of the molecules and mechanisms that regulate Ca(2+) homeostasis in this unique Ca(2+) signaling system. |
23260015 | Individual differences in recovery from traumatic fear. | Although exposure to major psychological trauma is unfortunately common, risk for related neuropsychiatric conditions, such as post-traumatic stress disorder (PTSD), varies greatly among individuals. Fear extinction offers a tractable and translatable behavioral readout of individual differences in learned recovery from trauma. Studies in rodent substrains and subpopulations are providing new insights into neural system dysfunctions associated with impaired fear extinction. Rapid progress is also being made in identifying key molecular circuits, epigenetic mechanisms, and gene variants associated with differences in fear extinction. Here, we discuss how this research is informing understanding of the etiology and pathophysiology of individual differences in risk for trauma-related anxiety disorders, and how future work can help identify novel diagnostic biomarkers and pharmacotherapeutics for these disorders. |
23260017 | Genetic associations of test-day fat:protein ratio with milk yield, fertility, and udder health traits in Nordic Red cattle. | Interest is growing in finding indicator traits for the evaluation of nutritional or tissue energy status of animals directly at the individual animal level. The development and subsequent use of such traits in practice demands a clear understanding of the genetic and phenotypic associations with the various production and functional traits. In this study, the relationships during lactation between milk fat:protein ratio (FPR) and production and functional traits were estimated for Nordic Red cattle, in which published information is scarce. The objectives of this study were to estimate genetic associations of FPR with milk yield (MY), fertility, and udder health traits during different stages of lactation. Traits included in the analyses were MY, 4 fertility traits-days from calving to insemination (DFI), days open (DO), number of inseminations (NI), and nonreturn rate to 56 d (NRR)-and 2 udder health traits-test-day somatic cell score (SCS) and clinical mastitis (CM). Data were from a total of 22,422 first-lactation cows. Random regression models were used to estimate genetic parameters and associations between traits. The mean FPR in first-lactation cows was 1.28 and ranged from 1.25 to 1.45. During first lactation, the heritability of FPR ranged from 0.14 to 0.25. Genetic correlations between FPR and MY in early lactation (until 50 d in milk) were positive and ranged from 0.05 to 0.22; later in lactation, they were close to zero or negative, indicating that cows may have come out of the negative state of energy balance. The strength of genetic associations between FPR and fertility traits varied during lactation. In early lactation, correlations between FPR and the interval fertility traits DFI and DO were positive and ranged from 0.14 to 0.28. Genetic correlations between FPR and the udder health traits SCS and CM in early lactation ranged from 0.09 to 0.20. Milk fat:protein ratio is a heritable trait and easily available from routine milk-recording schemes. It can be used as a low-cost monitoring tool of poor health and fertility in the most critical phases of lactation and as an important indicator trait to improve robustness in dairy cows through selection. |
23260014 | Microglia: actively surveying and shaping neuronal circuit structure and function. | The traditional role of microglia has been in brain infection and disease, phagocytosing debris and secreting factors to modify disease progression. Recent evidence extends their role to healthy brain homeostasis, including the regulation of cell death, synapse elimination, neurogenesis, and neuronal surveillance. These actions contribute to the maturation and plasticity of neural circuits that ultimately shape behavior. Here we review microglial contributions to the development, plasticity, and maintenance of neural circuits with a focus on interactions with synapses. We introduce this topic by reviewing recent studies on the migration and proliferation of microglia within the brain, and conclude with the proposal that microglia dysfunction may adversely affect brain function, and thereby contribute to the development of psychiatric and neurological disorders. |
23260013 | Determining adult type 2 diabetes-related health care needs in an indigenous population from rural Guatemala: a mixed-methods preliminary study. | In Guatemala, diabetes is an emerging public health concern. Guatemala has one of the largest indigenous populations in Latin America, and this population frequently does not access the formal health care system. Therefore, knowledge about the emergence of diabetes in this population is limited. Interview participants (n=23) were recruited from a convenience sample of indigenous adults with type 2 diabetes at one rural diabetes clinic in Guatemala. A structured interview was used to assess knowledge about diabetes and its complications; access to diabetes-related health care and treatment; dietary and lifestyle changes; and family and social supports for individuals living with diabetes. Interviews were supplemented with two group interviews with community leaders and health care providers. Thematic analysis was used to produce insights into diabetes knowledge, attitudes, and practices. In addition, a chart review of the clinic's electronic medical record identified all adult patients (n=80) presenting in one calendar year for a first-time diabetic consultation. Sociodemographic and clinical variables were extracted and summarized from these records. Salient demographic factors in both the structured interview and chart review samples included low educational levels and high indigenous language preference. In the interview sample, major gaps in biomedical knowledge about diabetes included understanding the causes, chronicity, and long-term end-organ complications of diabetes. Medication costs, medical pluralism, and limited social supports for dietary and lifestyles changes were major practical barriers to disease management. Quantitative data from medical records review revealed high rates of poor glycemic control, overweight and obesity, and medication prescription. This study provides a preliminary sketch of type 2 diabetes in an indigenous Guatemalan population. Combined qualitative and quantitative data point towards particular needs for implementation and future research, including the need to address gaps in diabetes knowledge, to improve social support systems, and to address the cost barriers associated with disease treatment. |
23260012 | Structural analysis of the genome of breast cancer cell line ZR-75-30 identifies twelve expressed fusion genes. | It has recently emerged that common epithelial cancers such as breast cancers have fusion genes like those in leukaemias. In a representative breast cancer cell line, ZR-75-30, we searched for fusion genes, by analysing genome rearrangements. We first analysed rearrangements of the ZR-75-30 genome, to around 10kb resolution, by molecular cytogenetic approaches, combining array painting and array CGH. We then compared this map with genomic junctions determined by paired-end sequencing. Most of the breakpoints found by array painting and array CGH were identified in the paired end sequencing-55% of the unamplified breakpoints and 97% of the amplified breakpoints (as these are represented by more sequence reads). From this analysis we identified 9 expressed fusion genes: APPBP2-PHF20L1, BCAS3-HOXB9, COL14A1-SKAP1, TAOK1-PCGF2, TIAM1-NRIP1, TIMM23-ARHGAP32, TRPS1-LASP1, USP32-CCDC49 and ZMYM4-OPRD1. We also determined the genomic junctions of a further three expressed fusion genes that had been described by others, BCAS3-ERBB2, DDX5-DEPDC6/DEPTOR and PLEC1-ENPP2. Of this total of 12 expressed fusion genes, 9 were in the coamplification. Due to the sensitivity of the technologies used, we estimate these 12 fusion genes to be around two-thirds of the true total. Many of the fusions seem likely to be driver mutations. For example, PHF20L1, BCAS3, TAOK1, PCGF2, and TRPS1 are fused in other breast cancers. HOXB9 and PHF20L1 are members of gene families that are fused in other neoplasms. Several of the other genes are relevant to cancer-in addition to ERBB2, SKAP1 is an adaptor for Src, DEPTOR regulates the mTOR pathway and NRIP1 is an estrogen-receptor coregulator. This is the first structural analysis of a breast cancer genome that combines classical molecular cytogenetic approaches with sequencing. Paired-end sequencing was able to detect almost all breakpoints, where there was adequate read depth. It supports the view that gene breakage and gene fusion are important classes of mutation in breast cancer, with a typical breast cancer expressing many fusion genes. |
23259992 | Impact of intestinal PepT1 on the kinetics and dynamics of N-formyl-methionyl-leucyl-phenylalanine, a bacterially-produced chemotactic peptide. | The primary purpose of this study was to evaluate the intestinal permeability (P(eff)) of N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe), a bacterially derived chemotactic tripeptide, in the duodenum, jejunum, ileum, and colon of wild-type and PepT1 knockout mice. A secondary purpose was to determine if the presence of intestinal PepT1 translated into fMet-Leu-Phe directed neutrophil migration in these animals. Using an in situ single pass perfusion technique, the P(eff) of [(3)H]fMet-Leu-Phe was substantially reduced in the duodenum, jejunum, and ileum of PepT1 knockout mice as compared to wild-type animals. In contrast, the P(eff) of [(3)H]fMet-Leu-Phe in colon was unchanged between genotypes and about 5% of that in small intestine. Jejunal uptake of [(3)H]fMet-Leu-Phe was specific for PepT1 and saturable with an intrinsic K(0.5) of 1.6 mM. The peptide/histidine transporters PhT1 and PhT2 were not involved in [(3)H]fMet-Leu-Phe uptake. Myeloperoxidase activity (a measure of neutrophil migration) was significantly increased following 4 h perfusions of 10 μM fMet-Leu-Phe in the jejunum of wild-type mice and was abolished by 50 mM glycylglycine; no change was observed in the jejunum of PepT1 knockout mice. Likewise, fMet-Leu-Phe perfusions had no effect on myeloperoxidase activity in the colon of either genotype. In conclusion, these findings demonstrated that PepT1 had a major influence on the permeability of fMet-Leu-Phe in duodenum, jejunum, and ileum in wild-type mice and on inflammatory response in intestinal regions that expressed PepT1. |
23259991 | Making "ethical safe space" in the translation of contested knowledge: the role of community debate in defining end-of-life decision ethics. | The objectives of this article are, first, to document a unique process of research knowledge translation (KT), which the authors describe as the creation of "ethical safe space," and, second, to document the narratives of forum participants and describe their interaction in a dialogue about vulnerability, the authority of physicians, and the perspective of people with disabilities on the policy. Narrative data from qualitative interviews with individual key informants and focus groups were used to identify speakers with specific expertise on policy, disability perspectives, and bioethical issues, who were invited to participate in the Forum on Ethical Safe Space. The planning workgroup adopted a model for enabling representative participation in the public forum designed to reduce the impact of physical, sensory, financial, language, and professional status barriers. Using the transcripts and keynote speakers' printed texts, primary themes and patterns of interaction were identified reflecting the alternative perspectives. Through the development of a workshop on ethical, legal, and disability-related implications of professional policy guidelines developed by the College of Physicians and Surgeons of Manitoba, we provided a qualitative analysis of the discourse involving experts and disability community members supporting alternative positions on the impact of the policy statement, and discuss ethical, legal, and disability rights issues identified in the public debate. Contested policy and ethical frameworks for making decisions about withdrawing and withholding life supporting treatment may influence both the perspectives of palliative care providers and patients referred to palliative care facilities. An innovative model for KT using a public forum that enabled stakeholders with conflicting perspectives to engage with ethical and professional policy issues asserting the physician's authority in contested decisions involving withdrawing or withholding life-supporting treatment, was a successful way to engage stakeholders supporting alternative positions on the impact of the policy statement and to discuss ethical, legal, and disability rights issues identified in the public debate. Discussion during the forum revealed several benefits of creating ethical safe space. This model of workshop allows space for participation of stakeholders, who might not otherwise be able to interact in the same forum, to articulate their perspectives and debate with other presenters and audience members. Participants at the forum spoke of the creation of ethical safe space as a starting point for more dialogue on the issues raised by the policy statement. The forum was, therefore, seen as a potential starting point for building conversation that would facilitate revising the policy with broader consultation on its legal and ethical validity. |
23259989 | Iron overload impairs proliferation of erythroid progenitors cells (BFU-E) from patients with myelodysplastic syndromes. | In patients with myelodysplastic syndromes (MDS) iron overload caused by long-term red blood cell transfusions is an important factor for comorbidity especially in low-risk MDS. In this report we present the results of a comparative study based on colony formation assays of hematopoietic cells in MDS patients with and without iron overload. We demonstrate that iron overload suppresses the proliferation of erythroid progenitors cells (BFU-E), while the myeloid compartment (CFU-GM) was not found to be affected. Even patients with slightly elevated ferritin values show an impaired proliferation capacity in comparison to patients with normal ferritin levels. Furthermore, we show that this negative impact is reversible by sufficient iron chelation therapy. |
23259988 | Influence of time to complete remission and duration of all-trans retinoic acid therapy on the relapse risk in patients with acute promyelocytic leukemia receiving AIDA protocols. | Despite the impressive results obtained with standard chemotherapy, approximately 20% of acute promyelocytic leukemia (APL) patients undergo disease relapse thereby requiring salvage therapy. Few data is available on long-term prognosis in relation to time to complete remission (CR): we reviewed 142 patients treated with AIDA protocols and we found that 42 out of 142 (29.6%) patients achieved CR after 35 days (median time, 42 days). No significant differences in presenting features, including FAB subtype, type of PML/RARA transcript and relapse risk at presentation between the two patient groups achieving CR > or <35 days were revealed, except for male sex and older age that were significantly associated with delayed CR. Rate of relapse was 31% in patients with delayed CR compared to 17% in the group of patients who achieved CR<35 days (p=0.001), with a 5-year CIR of 29.6% compared to 12% (p=0.03). APL patients with delayed CR should be more closely monitored during follow-up for early identification of relapse and prompt administration of pre-emptive salvage therapy. |
23259987 | Correlation between cytokine levels and changes in fatigue and quality of life in patients with acute myeloid leukemia. | Cancer-related fatigue (CRF) is a major problem in patients with acute myeloid leukemia (AML) and may be mediated by circulating cytokines. We examined this relationship in 74 adult AML patients before and after the first cycle of induction chemotherapy. Plasma levels of 13 cytokines were measured via electrochemiluminescence. At baseline, potentially clinically important (r>0.30) correlations were seen between tumor necrosis factor (TNF)-α and fatigue (r=-0.336, p=0.017). Over time, correlations with fatigue were noted with TNF-α (r=-0.341, p=0.006) and interferon-inducible protein (IP)-10 (r=0.353, p=0.005). The link between IP-10 and fatigue is novel, implicating CXC chemokine pathways for CRF in hematologic malignancies. |
23259986 | Endogenous IL-8 acts as a CD16 co-activator for natural killer-mediated anti-CD20 B cell depletion in chronic lymphocytic leukemia. | Rituximab (RTX, anti-CD20 antibody) combined with chemotherapy is currently standard treatment for chronic lymphocytic leukemia (CLL). Serum level of IL-8 is a prognostic factor for CLL that correlates with disease stage. We investigated whether endogenous IL-8 affects RTX or Obinutuzumab (GA101) B-leukemic depletion mediated by natural killers (NK). Using whole peripheral blood lymphocytes from untreated CLL patients, RTX, but most significantly GA101, were effective in B-cell depletion and NK activation. IL-8 inhibition completely inhibited B-cell depletion by RTX and reduced GA101-induced B-cell depletion. Altogether results underline IL-8 as an endogenous NK co-activator and confirm GA101 therapeutic potential for CLL treatment. |
23259985 | Bioreducible polymers as a determining factor for polyplex decomplexation rate and transfection. | Polyplex formation (complexation) and gene release from the polyplexes (decomplexation) are major events in polymeric gene delivery; however, the effect of the decomplexation rate on transfection has been rarely investigated. This study employed mixed polymers of poly((L)-lysine) (PLL: MW ~7.4 kDa) and reducible PLL (RPLL) (MW ~6.7 kDa) to design decomplexation rate-controllable PLL(100-x)RPLL(x)/pDNA complexes (PRL(x) polyplexes). The transfection efficiency of a model gene (luciferase) in MCF7 and HEK293 cell lines increased with increasing x (RPLL content) in the PRL(x) polyplexes until peaking at x = 2.5 and 10, respectively, after which point transfection efficiency declined rapidly. In MCF7 cells, PRL(2.5) polyplex produced 3 or 223 times higher gene expression than PLL or RPLL polyplexes, respectively. Similarly, the transfection efficiency of PRL(10) polyplex-transfected HEK293 cells was 3.8 or 67 times higher than that of PLL or RPLL polyplexes, respectively. The transfection results were not apparently related to the particle size, surface charge, complexation/compactness, cellular uptake, or cytotoxicity of the tested polyplexes. However, the decomplexation rate varied by RPLL content in the polyplexes, which in turn influenced the gene transfection. The nuclear localization of pDNA delivered by PRL(x) polyplexes showed a similar trend to their transfection efficiencies. This study suggests that an optimum decomplexation rate may result in high nuclear localization of pDNA and transfection. Understanding in decomplexation and intracellular localization of pDNA may help develop more effective polyplexes. |
23259984 | Vaccination and tick-borne encephalitis, central Europe. | Tick-borne encephalitis (TBE) is a substantial public health problem in many parts of Europe and Asia. To assess the effect of increasing TBE vaccination coverage in Austria, we compared incidence rates over 40 years for highly TBE-endemic countries of central Europe (Czech Republic, Slovenia, and Austria). For all 3 countries we found extensive annual and longer range fluctuations and shifts in distribution of patient ages, suggesting major variations in the complex interplay of factors influencing risk for exposure to TBE virus. The most distinctive effect was found for Austria, where mass vaccination decreased incidence to ≈16% of that of the prevaccination era. Incidence rates remained high for the nonvaccinated population. The vaccine was effective for persons in all age groups. During 2000-2011 in Austria, ≈4,000 cases of TBE were prevented by vaccination. |
23259983 | On multiple scattering in acoustic media: a deterministic Ray Tracing method for random structures. | The paper is devoted to computer and experimental simulation of US (ultrasonic) signal propagation in acoustic solids with micro-structure. Any change in the percentage of flaws or pores influences considerably the value of the ultrasonic wave speed. The theoretical analysis is based upon the Ray Tracing algorithm. We calculate numerically the full path of each ray from the transmitter to the receiver, in its multiple reflections between the surfaces of the internal obstacles. The natural experiments are performed in a water basin with some arrays of equal metallic round rods. This simulates the US evaluation of the mechanical properties of concrete. The computer modeling allows us to construct the envelope of the US signal registered at the receiving transducer. Then we simulate the dependence of the wave speed versus porosity. There is a sufficiently good accordance between numerical and experimental results. |
23259981 | Comparison of cardiac displacement and strain imaging using ultrasound radiofrequency and envelope signals. | Echocardiographic strain imaging is a promising new method for quantifying and displaying the health of cardiac muscle. Accurate regional myocardial function analysis requires high spatial and temporal resolution in addition to fidelity to the underlying deformation. However, all current clinical approaches use speckle-tracking algorithms applied to B-mode images derived from envelope signals. Such approaches are inherently of lower spatial resolution, since they require larger data blocks for deformation tracking due to the absence of phase information. In this paper, we compare the strain estimation performance using B-mode, envelope and radiofrequency signals, utilizing data acquired from a uniformly elastic tissue mimicking phantom, cardiac simulation, and clinical in vivo data. Signal-to-noise ratio improvements using radiofrequency signals for linear and phased array geometries were 5.80 dB and 9.48 dB over that obtained with envelope signals (at peak strain) in phantom studies, respectively. Cardiac simulation studies demonstrate that when averaged over the two cardiac cycles, the mean standard deviation of estimated strain using envelope signals from two of the six segments for a short-axes view (anterior and anterolateral) were 48% and 44% higher than that obtained using radiofrequency signals. These segments were chosen since one was along while the other was situated lateral to the beam propagation direction. In a similar manner, in vivo analysis on a volunteer also indicate that the standard deviation of the estimated strain using B-mode and envelope signals were 16% and 42% higher than that obtained using radiofrequency signals in the anteroseptal segment, and 45% and 27% in the anterior segment. These results demonstrate the significant reduction in the variability of strain estimated along with improvements in the spatial resolution and signal-to-noise ratios obtained using radiofrequency signals. |
23259982 | Examination of the epicentral waveform for laser ultrasound in the melting regime. | A laser ultrasonic source just below the ablation regime is examined by recording an epicentral waveform in a high purity tungsten sample. Using pulse energy as a parameter, a slight delay in the shear wave arrival time is observed upon transition to the melting regime. This phenomenon is attributed to a change in character of the ultrasonic source. In the thermoelastic regime, shear waves are generated by mode conversion at the sample surface of longitudinal waves emanating from subsurface sources. Just above the melting threshold, a molten pool forms in the center of the generation volume. Shear waves are not supported by the molten pool. As a result, shear waves generated from off-axis thermoelastic sources are weighted more heavily. This results in a delay of the shear wave arrival time. |
23259980 | Study of the resonances of periodic plane media immersed in water: theory and experiment. | The paper deals with the study of the resonances of 1D periodic media composed of N elementary cells formed with two perfectly bonded layers which exhibit a high acoustic impedance contrast. In the case of a periodic bilayer structure constituted of a fluid layer and an elastic plate, it was shown in previous theoretical works that additional modes appear compared to those of a single plate. These are called structure modes. At low frequency, the so-called vertical modes are found. Approximate expressions of their cut-off frequencies are given and their numerical values match with the exact ones. At high frequency, the Lamb type modes are degenerated and modes in the fluid layers are also observed. Preliminary experimental results have already proved the existence of such phenomena for one and two periods. In our work, an experimental validation has been performed in the case of N periods made with a glass isotropic elastic plate and a water fluid layer, where the number N ranges from two to five. A good agreement is shown compared to theoretical works. |
23259979 | Computational modeling and experimental studies of the dynamic performance of ultrasonic horn profiles used in plastic welding. | Ultrasonic horns are tuned components designed to vibrate in a longitudinal mode at ultrasonic frequencies. Reliable performance of such horns is normally decided by the uniformity of vibration amplitude at the working surface and the stress developed during loading condition. The horn design engineer must pay particular attention to designing a tool that will produce the desired amplitude without fracturing. The present work discusses horn configurations which satisfy these criteria and investigates the design requirements of horns in ultrasonic system. Different horn profiles for ultrasonic welding of thermoplastics have been characterized in terms of displacement amplitude and von-Mises stresses using modal and harmonic analysis. To validate the simulated results, five different horns are fabricated from Aluminum, tested and tuned to the operating frequency. Standard ABS plastic parts are welded using these horns. Temperature developed during the welding of ABS test parts using different horns is recorded using sensors and National Instruments (NIs) data acquisition system. The recorded values are compared with the predicted values. Experimental results show that welding using a Bezier horn has a high interface temperature and the welded joints had higher strength as compared to the other horn profiles. |
23259978 | Effects of viscous liquid on SH-SAW in layered magnetoelectric structures. | We investigate analytically shear horizontal surface acoustic wave (SH-SAW) propagation in layered magnetoelectric structures loaded with viscous liquid, which involves a thin piezomagnetic layer bonded perfectly to an unbounded piezoelectric substrate. The dispersive relations are obtained and the effects of liquid viscosity on the phase velocity and attenuation of the waves are analyzed and discussed. From the results we can find that the effects of the liquid viscosity on the properties of SH-SAW are remarkable. The phase velocity decreases with increase of the viscous coefficient, or with increase of the frequency, and the attenuation increases with the frequency of the waves and the liquid viscosity, respectively. The relationship between attenuation and frequency or viscosity is nonlinear, but the former is a concave curve, whereas the latter is a convex curve. The attenuation decreases with the piezomagnetic coefficient, and increases obviously with the thickness of the layer. The analytical method and the results are useful for the design of acoustic wave devices based on magnetoelectric materials for liquid phase application, which could be resonated by either magnetic or electric fields. |
23259976 | Multidetector-row computed tomography for evaluating the branching angle of the celiac artery: a descriptive study. | We performed this study in order to investigate the shape of the origin of the celiac artery in maximum intensity projection (MIP) using routine 64 multidetector-row computed tomography (MDCT) data in order to plan for the implantation of an intra-arterial hepatic port system. A total of 1,104 patients with hepatocellular carcinoma were assessed with MDCT. In the definition of the branching angle, the anterior side of the abdominal aorta was considered the baseline, and the cranial and caudal sides were designated as 0 and 180 degrees, respectively. The angles between 0 and 90 degrees and between 90 and 180 degrees from the cranial side were considered upward and downward, respectively, and the branching angle of the celiac artery was classified every 30 degrees. The subclavian arterial route was used for the implantation of an intra-arterial hepatic port system in patients with branching angles of 150 degrees or more (sharp downward). The median branching angle was (median ± standard deviation) 135 ± 23 (range, 51-174) degrees. The branching was upward in 77 patients (7%) and downward in 1,027 patients (93%). The branching was downward with an angle of 120 to 150 degrees in most patients (n = 613). The branching was sharply downward with an angle of 150 degrees or more in 177 patients (16%). A total of 10 patients were referred for interventional placement of an intra-arterial hepatic port system. The subclavian arterial route was used for implantation of an intra-arterial hepatic port system in 2 patients with sharp downward branching. The branching angle of the celiac artery can be easily determined by the preparation of MIP images from routine MDCT data. MIP may provide useful information for the selection of the catheter insertion route in order to avoid a sharp branching angle of the celiac artery. |
23259975 | Communicating with the public following radiological terrorism: results from a series of focus groups and national surveys in Britain and Germany. | Incidents involving the exposure of large numbers of people to radiological material can have serious consequences for those affected, their community and wider society. In many instances, the psychological effects of these incidents have the greatest impact. People fear radiation and even incidents which result in little or no actual exposure have the potential to cause widespread anxiety and behavior change. The aim of this study was to assess public intentions, beliefs and information needs in the UK and Germany in response to a hidden radiological exposure device. By assessing how the public is likely to react to such events, strategies for more effective crisis and risk communication can be developed and designed to address any knowledge gaps, misperceptions and behavioral responses that are contrary to public health advice. This study had three stages. The first stage consisted of focus groups which identified perceptions of and reactions to a covert radiological device. The incident was introduced to participants using a series of mock newspaper and broadcast injects to convey the evolving scenario. The outcomes of these focus groups were used to inform national telephone surveys, which quantified intended behaviors and assessed what perceptions were correlated with these behaviors. Focus group and survey results were used to develop video and leaflet communication interventions, which were then evaluated in a second round of focus groups. In the first two stages, misperceptions about the likelihood and routes of exposure were associated with higher levels of worry and greater likelihood of engaging in behaviors that might be detrimental to ongoing public health efforts. The final focus groups demonstrated that both types of misunderstanding are amenable to change following targeted communication. Should terrorists succeed in placing a hidden radiological device in a public location, then health agencies may find that it is easier to communicate effectively with the public if they explicitly and clearly discuss the mechanisms through which someone could be affected by the radiation and the known geographical spread of any risk. Messages which explain how the risk from a hidden radiological device "works" should be prepared and tested in advance so that they can be rapidly deployed if the need arises. |
23259973 | Coordinated development of voltage-gated Na+ and K+ currents regulates functional maturation of forebrain neurons derived from human induced pluripotent stem cells. | Like embryonic stem (ES) cells, human induced pluripotent stem (hiPS) cells can differentiate into neuronal cells. However, it is unclear how their exquisite neuronal function is electrophysiologically coordinated during differentiation and whether they are functionally identical to human ES cell-derived neurons. In this study, we differentiated hiPS and ES cells into pyramidal-like neurons and conducted electrophysiological characterization over the 4-week terminal differentiation period. The human neuron-like cells express forebrain pyramidal cell markers NeuN, neurofilament, the microtubule-associated protein 2 (MAP2), the paired box protein Pax-6 (PAX6), Tuj1, and the forkhead box protein G1 (FoxG1). The size of developing neurons increased continuously during the 4-week culture, and cell-resting membrane potentials (RMPs) underwent a negative shift from -40 to -70 mV. Expression of the muscarinic receptor-modulated K(+) currents (IM) participated in the development of cell RMPs and controlled excitability. Immature neurons at week 1 could only fire abortive action potentials (APs) and the frequency of AP firing progressively increased with neuronal maturation. Interestingly, the developmental change of voltage-gated Na(+) current (INa) did not correlate with the change in the AP firing frequency. On the other hand, the transient outward K(+) current (IA), but not the delayed rectifier current (IK) contributed to the high frequency firing of APs. Synaptic activities were observed throughout the 4-week development. These morphological and electrophysiological features were almost identical between iPS and ES cell-derived neurons. This is the first systematic investigation showing functional evidence that hiPS cell-derived neurons possess similar neuronal activities as ES cell-derived neurons. These data support that iPS cell-derived neural progenitor cells have the potential for replacing lost neurons in cell-based therapy. |
23259974 | Characterization of wood plastic composites made from landfill-derived plastic and sawdust: volatile compounds and olfactometric analysis. | Application of wood plastic composites (WPCs) obtained from recycled materials initially intended for landfill is usually limited by their composition, mainly focused on release of volatile organic compounds (VOCs) which could affect quality or human safety. The study of the VOCs released by a material is a requirement for new composite materials. Characterization and quantification of VOCs of several WPC produced with low density polyethylene (LDPE) and polyethylene/ethylene vinyl acetate (PE/EVA) films and sawdust were carried out, in each stage of production, by solid phase microextraction in headspace mode (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). An odor profile was also obtained by HS-SPME and GC-MS coupled with olfactometry analysis. More than 140 compounds were observed in the raw materials and WPC samples. Some quantified compounds were considered WPC markers such as furfural, 2-methoxyphenol, N-methylphthalimide and 2,4-di-tert-butylphenol. Hexanoic acid, acetic acid, 2-methoxyphenol, acetylfuran, diacetyl, and aldehydes were the most important odorants. None of the VOCs were found to affect human safety for use of the WPC. |
23259972 | Isolation, structure, and biological activity of Phaeofungin, a cyclic lipodepsipeptide from a Phaeosphaeria sp. Using the Genome-Wide Candida albicans Fitness Test. | Phaeofungin (1), a new cyclic depsipeptide isolated from Phaeosphaeria sp., was discovered by application of reverse genetics technology, using the Candida albicans fitness test (CaFT). Phaeofungin is comprised of seven amino acids and a β,γ-dihydroxy-γ-methylhexadecanoic acid arranged in a 25-membered cyclic depsipeptide. Five of the amino acids were assigned with d-configurations. The structure was elucidated by 2D-NMR and HRMS-MS analysis of the natural product and its hydrolyzed linear peptide. The absolute configuration of the amino acids was determined by Marfey's method by complete and partial hydrolysis of 1. The CaFT profile of the phaeofungin-containing extract overlapped with that of phomafungin (3), another structurally different cyclic lipodepsipeptide isolated from a Phoma sp. using the same approach. Comparative biological characterization further demonstrated that these two fungal lipodepsipeptides are functionally distinct. While phomafungin was potentiated by cyclosporin A (an inhibitor of the calcineurin pathway), phaeofungin was synergized with aureobasidin A (2) (an inhibitor of the sphingolipid biosynthesis) and to some extent caspofungin (an inhibitor of glucan synthase). Furthermore, phaeofungin caused ATP release in wild-type C. albicans strains but phomafungin did not. It showed modest antifungal activity against C. albicans (MIC 16-32 μg/mL) and better activity against Aspergillus fumigatus (MIC 8-16 μg/mL) and Trichophyton mentagrophytes (MIC 4 μg/mL). The linear peptide was inactive, suggesting that the macrocyclic depsipeptide ring is essential for target engagement and antifungal activity. |
23259971 | Endotoxin markers in bronchoalveolar lavage fluid of patients with interstitial lung diseases. | Exposure to inhaled endotoxins (lipopolysaccharides, LPS) of Gram-negative bacteria commonly found in indoor environments and assessed in secondary tobacco smoke, has been associated with airway inflammation and asthma exacerbation. The bronchoalveolar lavage fluid (BALf) from patients with interstitial lung diseases (sarcoidosis, lung fibrosis, smoking-related ILD, eosinophilic disorders) was analyzed for the markers of lipopolysaccharide (LPS, endotoxin). BALf was obtained from patients with diffuse lung diseases: idiopathic pulmonary fibrosis (n = 42), sarcoidosis (n = 22), smoking-related-ILD (n = 11) and eosinophilic disorders (n = 8). Total cell count and differential cell count were performed. In addition, samples were analyzed for 3-hydroxy fatty acids (3-OHFAs) of 10-18 carbon chain lengths, as markers of LPS, by gas chromatography-tandem mass spectrometry. The highest LPS concentration was found in patients with eosinophilic disorders and the lowest in patients with sarcoidosis (p< 0.05) followed by the lung fibrosis and the sr-ILD patients. The difference between LPS in BALf with extremely high eosinophil proportion (> 25%) and those with lower proportion was also significant (p = 0.014). A significant correlation was found between LPS and eosinophils, but not between LPS and lymphocytes, neutrophils, or macrophages count. A positive relationship of LPS and eosinophilic pulmonary disorders may be linked to a persistent eosinophil activation mediated by Th2 pathway: chronic endotoxin exposure would intensify Th2 pathway resulting in fibrosis and, at the same time, eosinophil stimulation, and hence in eosinophilic pulmonary disorders. |
23259970 | Evolutionary relationships between miRNA genes and their activity. | The emergence of vertebrates is characterized by a strong increase in miRNA families. MicroRNAs interact broadly with many transcripts, and the evolution of such a system is intriguing. However, evolutionary questions concerning the origin of miRNA genes and their subsequent evolution remain unexplained. In order to systematically understand the evolutionary relationship between miRNAs gene and their function, we classified human known miRNAs into eight groups based on their evolutionary ages estimated by maximum parsimony method. New miRNA genes with new functional sequences accumulated more dynamically in vertebrates than that observed in Drosophila. Different levels of evolutionary selection were observed over miRNA gene sequences with different time of origin. Most genic miRNAs differ from their host genes in time of origin, there is no particular relationship between the age of a miRNA and the age of its host genes, genic miRNAs are mostly younger than the corresponding host genes. MicroRNAs originated over different time-scales are often predicted/verified to target the same or overlapping sets of genes, opening the possibility of substantial functional redundancy among miRNAs of different ages. Higher degree of tissue specificity and lower expression level was found in young miRNAs. Our data showed that compared with protein coding genes, miRNA genes are more dynamic in terms of emergence and decay. Evolution patterns are quite different between miRNAs of different ages. MicroRNAs activity is under tight control with well-regulated expression increased and targeting decreased over time. Our work calls attention to the study of miRNA activity with a consideration of their origin time. |
23259969 | Patient characteristics associated with hospitalisations for ambulatory care sensitive conditions in Victoria, Australia. | Ambulatory Care Sensitive Conditions (ACSCs) are those for which hospitalisation is thought to be avoidable with the application of preventive care and early disease management, usually delivered in a primary care setting. ACSCs are used extensively as indicators of accessibility and effectiveness of primary health care. We examined the association between patient characteristics and hospitalisation for ACSCs in the adult and paediatric population in Victoria, Australia, 2003/04. Hospital admissions data were merged with two area-level socioeconomic indexes: Index of Socio-Economic Disadvantage (IRSED) and Accessibility/Remoteness Index of Australia (ARIA). Univariate and multiple logistic regressions were performed for both adult (age 18+ years) and paediatric (age <18 years) groups, reporting odds ratios (OR) and 95% confidence intervals (CI) for a number of predictors of ACSCs admissions compared to non-ACSCs admissions. Predictors were much more strongly associated with ACSCs admissions compared to non-ACSCs admissions in the adult group than for the paediatric group with the exception of rurality. Significant adjusted ORs in the adult group were 1.06, 1.15, 1.13, 1.06 and 1.11 for sex, rurality, age, IRSED and ARIA variables, and 1.34, 1.04 and 1.09 in the paediatric group for rurality, IRSED and ARIA, respectively. Disadvantaged paediatric and adult population experience more need of hospital care for ACSCs. Access barriers to primary care are plausible causes for the observed disparities. Understanding the characteristics of individuals experiencing access barriers to primary care will be useful for developing targeted interventions meeting the unique ambulatory needs of the population. |
23259963 | Recruitment and retention of general practitioners in rural Canada and Australia: a review of the literature. | Both Canada and Australia are facing severe shortages of primary health workers, and these shortages are exacerbated in rural and remote communities. This literature review highlights similarities and explores the factors that serve to attract and retain family practitioners in underserved regions of Canada and Australia. We used MEDLINE on OvidSP to review the literature between Jan. 1, 2000, and June 30, 2012. We excluded sources if the primary objective did not consider recruitment or retention of general practitioners. We found a total of 114 sources, 28 of which were excluded, leaving 86 sources for review. We organized results according to 5 life stages of family physicians in rural practice and graded the literature according to the strength of the methodology and the relevance of the findings. We chronologically categorized Canadian and Australian literature that discussed recruitment and retention of family practitioners into rural practice. Various factors that pertain to each life stage of a family physician have been shown to positively correlate with the eventual decision to commence and remain practising in rural areas. Training programs should be better structured to attract candidates who are more likely to enter rural practice. Policy-makers should be mindful of these findings, because improvements in retention will deliver large financial savings. |
23259962 | Perceptions of medical school among high school students in southwestern Ontario. | Canadian medical students are more likely to come from urban and high-income areas and to have well-educated, professional parents. Physicians who grew up in rural areas are more likely to serve in rural and lower-income areas. We identify perceptions held by rural high school students regarding the affordability and attainability of a medical education. We distributed a survey to high school students who attended the MedQUEST Health Career Exploration Program in southwestern Ontario. The survey assessed socioeconomic background and perceived barriers to a medical education (including affordability as well as encouragement and discouragement from others). Of the 119 attendees, 106 (89.1%) completed the survey. Of the students who were interested in becoming physicians, most expected to fund their medical education through scholarships (56 [69.1%]), parental support (50 [61.7%]) or student employment (45 [55.6%]). However, less than half of all respondents (48 [45.3%]) provided reasonably correct estimates for annual medical tuition fees. If at least 1 parent had a postsecondary education, respondents were less likely to cite affordability as a barrier to attending medical school (p = 0.05). Although students interested in obtaining a medical education cited affordability as a potential barrier, many were not aware of the actual cost of attending medical school. We found an association between perceived affordability of medical school and parents' level of education. To define this relation further, research is needed to collect more accurate data on family income. Students may benefit from more information about funding opportunities for medical school. |
23259957 | Incorporation of monodisperse oligoethyleneglycol amino acids into anticonvulsant analogues of galanin and neuropeptide y provides peripherally acting analgesics. | Delivery of neuropeptides into the central and/or peripheral nervous systems supports development of novel neurotherapeutics for the treatment of pain, epilepsy and other neurological diseases. Our previous work showed that the combination of lipidization and cationization applied to anticonvulsant neuropeptides galanin (GAL) and neuropeptide Y (NPY) improved their penetration across the blood-brain barrier yielding potent antiepileptic lead compounds, such as Gal-B2 (NAX 5055) or NPY-B2. To dissect peripheral and central actions of anticonvulsant neuropeptides, we rationally designed, synthesized and characterized GAL and NPY analogues containing monodisperse (discrete) oligoethyleneglycol-lysine (dPEG-Lys). The dPEGylated analogues Gal-B2-dPEG(24), Gal-R2-dPEG(24) and NPY-dPEG(24) displayed analgesic activities following systemic administration, while avoiding penetration into the brain. Gal-B2-dPEG(24) was synthesized by a stepwise deprotection of orthogonal 4-methoxytrityl and allyloxycarbonyl groups, and subsequent on-resin conjugations of dPEG(24) and palmitic acids, respectively. All the dPEGylated analogues exhibited substantially decreased hydrophobicity (expressed as logD values), increased in vitro serum stabilities and pronounced analgesia in the formalin and carrageenan inflammatory pain assays following systemic administration, while lacking apparent antiseizure activities. These results suggest that discrete PEGylation of neuropeptides offers an attractive strategy for developing neurotherapeutics with restricted penetration into the central nervous system. |
23259956 | Fractionating human intelligence. | What makes one person more intellectually able than another? Can the entire distribution of human intelligence be accounted for by just one general factor? Is intelligence supported by a single neural system? Here, we provide a perspective on human intelligence that takes into account how general abilities or "factors" reflect the functional organization of the brain. By comparing factor models of individual differences in performance with factor models of brain functional organization, we demonstrate that different components of intelligence have their analogs in distinct brain networks. Using simulations based on neuroimaging data, we show that the higher-order factor "g" is accounted for by cognitive tasks corecruiting multiple networks. Finally, we confirm the independence of these components of intelligence by dissociating them using questionnaire variables. We propose that intelligence is an emergent property of anatomically distinct cognitive systems, each of which has its own capacity. |
23259955 | A continuous semantic space describes the representation of thousands of object and action categories across the human brain. | Humans can see and name thousands of distinct object and action categories, so it is unlikely that each category is represented in a distinct brain area. A more efficient scheme would be to represent categories as locations in a continuous semantic space mapped smoothly across the cortical surface. To search for such a space, we used fMRI to measure human brain activity evoked by natural movies. We then used voxelwise models to examine the cortical representation of 1,705 object and action categories. The first few dimensions of the underlying semantic space were recovered from the fit models by principal components analysis. Projection of the recovered semantic space onto cortical flat maps shows that semantic selectivity is organized into smooth gradients that cover much of visual and nonvisual cortex. Furthermore, both the recovered semantic space and the cortical organization of the space are shared across different individuals. |
23259954 | The postsaccadic unreliability of gain fields renders it unlikely that the motor system can use them to calculate target position in space. | Gain fields, the eye-position modulation of visual responses, are thought to provide a mechanism by which the motor system can accurately calculate target position in space despite a constantly moving eye. Current gain-field models assume that the modulation of visual responses by eye position is accurate at all times, even around the time of a saccade. Here, we show that for at least 150 ms after a saccade, gain fields in the lateral intraparietal area (LIP) are unreliable. The majority of LIP cells with steady-state gain fields reflect the presaccadic eye position. The remainder of the cells have responses that cannot be predicted by their steady-state gain fields. Nonetheless, a monkey's oculomotor performance is accurate during this time. These results suggest that current models built upon a simple gain-field algorithm cannot be used to calculate the position of a target in space that flashes briefly after a saccade. |
23259952 | Functional properties of cortical feedback projections to the olfactory bulb. | Sensory perception is not a simple feed-forward process, and higher brain areas can actively modulate information processing in "lower" areas. We used optogenetic methods to examine how cortical feedback projections affect circuits in the first olfactory processing stage, the olfactory bulb. Selective activation of back projections from the anterior olfactory nucleus/cortex (AON) revealed functional glutamatergic synaptic connections on several types of bulbar interneurons. Unexpectedly, AON axons also directly depolarized mitral cells (MCs), enough to elicit spikes reliably in a time window of a few milliseconds. MCs received strong disynaptic inhibition, a third of which arises in the glomerular layer. Activating feedback axons in vivo suppressed spontaneous as well as odor-evoked activity of MCs, sometimes preceded by a temporally precise increase in firing probability. Our study indicates that cortical feedback can shape the activity of bulbar output neurons by enabling precisely timed spikes and enforcing broad inhibition to suppress background activity. |
23259953 | Hilar mossy cell degeneration causes transient dentate granule cell hyperexcitability and impaired pattern separation. | Although excitatory mossy cells of the hippocampal hilar region are known to project both to dentate granule cells and to interneurons, it is as yet unclear whether mossy cell activity's net effect on granule cells is excitatory or inhibitory. To explore their influence on dentate excitability and hippocampal function, we generated a conditional transgenic mouse line, using the Cre/loxP system, in which diphtheria toxin receptor was selectively expressed in mossy cells. One week after injecting toxin into this line, mossy cells throughout the longitudinal axis were degenerated extensively, theta wave power of dentate local field potentials increased during exploration, and deficits occurred in contextual discrimination. By contrast, we detected no epileptiform activity, spontaneous behavioral seizures, or mossy-fiber sprouting 5-6 weeks after mossy cell degeneration. These results indicate that the net effect of mossy cell excitation is to inhibit granule cell activity and enable dentate pattern separation. |
23259951 | Cortical feedback control of olfactory bulb circuits. | Olfactory cortex pyramidal cells integrate sensory input from olfactory bulb mitral and tufted (M/T) cells and project axons back to the bulb. However, the impact of cortical feedback projections on olfactory bulb circuits is unclear. Here, we selectively express channelrhodopsin-2 in olfactory cortex pyramidal cells and show that cortical feedback projections excite diverse populations of bulb interneurons. Activation of cortical fibers directly excites GABAergic granule cells, which in turn inhibit M/T cells. However, we show that cortical inputs preferentially target short axon cells that drive feedforward inhibition of granule cells. In vivo, activation of olfactory cortex that only weakly affects spontaneous M/T cell firing strongly gates odor-evoked M/T cell responses: cortical activity suppresses odor-evoked excitation and enhances odor-evoked inhibition. Together, these results indicate that although cortical projections have diverse actions on olfactory bulb microcircuits, the net effect of cortical feedback on M/T cells is an amplification of odor-evoked inhibition. |
23259950 | Layer-specific targeting of direction-selective neurons in the zebrafish optic tectum. | Direction selectivity (DS) is an important neuronal property in the visual system, but how DS is generated beyond the retina remains controversial. Here, we report a close correspondence between the preferred direction (PD) and the morphology of DS cells in the optic tectum. Ca(2+) imaging in cells expressing the genetically encoded Ca(2+) indicator GCaMP3 and multiphoton-targeted patch-clamp recordings allowed us to compare structure and function in single neurons. The arbors of differently tuned cell types showed stereotypic differences in shape and laminar profile within the tectal neuropil. Excitatory synaptic inputs were directionally tuned and matched the PD of spike output in these cells, while inhibitory inputs were selective for nonpreferred directions. Functional Ca(2+) imaging in afferent axons showed a matching laminar distribution of DS presynaptic activity. Hence, different directions are represented in different layers, which suggests a simple mechanism for how tectal neurons acquire directional tuning in a nascent circuit. |
23259949 | Activity-dependent transcriptional regulation of M-Type (Kv7) K(+) channels by AKAP79/150-mediated NFAT actions. | M-type K(+) channels, encoded by KCNQ2-KCNQ5 genes, play key roles in regulation of neuronal excitability; however, less is known about the mechanisms controlling their transcriptional expression. Here, we discovered a mechanism regulating KCNQ2/3 transcriptional expression by neuronal activity in rodent neurons, involving activation of calcineurin and nuclear factor of activated T cell (NFAT) transcription factors, orchestrated by A kinase-anchoring protein (AKAP)79/150. The signal requires Ca(2+) influx through L-type Ca(2+) channels and both local and global Ca(2+) elevations. We postulate increased M-channel expression to act as a negative feedback to suppress neuronal hyperexcitability, demonstrated by profoundly upregulated KCNQ2/3 transcription in hippocampi from wild-type, but not AKAP150(-/-), mice after drug-induced seizures. Thus, we suggest a distinct role of AKAP79/150 and the complex it organizes in activity-dependent M-channel transcription, which may potentially serve throughout the nervous system to limit overexcitability associated with disease states such as epilepsy. |
23259948 | Genetic dissection of TAM receptor-ligand interaction in retinal pigment epithelial cell phagocytosis. | Although TAM receptor tyrosine kinases play key roles in immune regulation, cancer metastasis, and viral infection, the relative importance of the two TAM ligands-Gas6 and Protein S-has yet to be resolved in any setting in vivo. We have now performed a genetic dissection of ligand function in the retina, where the TAM receptor Mer is required for the circadian phagocytosis of photoreceptor outer segments by retinal pigment epithelial cells. This process is severely attenuated in Mer mutant mice, which leads to photoreceptor death. We find that retinal deletion of either Gas6 or Protein S alone yields retinae with a normal number of photoreceptors. However, concerted deletion of both ligands fully reproduces the photoreceptor death seen in Mer mutants. These results demonstrate that Protein S and Gas6 function as independent, bona fide Mer ligands, and are, to a first approximation, interchangeable with respect to Mer-driven phagocytosis in the retina. |
23259947 | Tangentially migrating neurons assemble a primary cilium that promotes their reorientation to the cortical plate. | In migrating neurons, the centrosome nucleates and anchors a polarized network of microtubules that directs organelle movements. We report here that the mother centriole of neurons migrating tangentially from the medial ganglionic eminence (MGE) assembles a short primary cilium and exposes this cilium to the cell surface by docking to the plasma membrane in the leading process. Primary cilia are built by intraflagellar transport (IFT), which is also required for Sonic hedgehog (Shh) signal transduction in vertebrates. We show that Shh pathway perturbations influenced the leading process morphology and dynamics of MGE cells. Whereas Shh favored the exit of MGE cells away from their tangential migratory paths in the developing cortex, cyclopamine or invalidation of IFT genes maintained MGE cells in the tangential paths. Our findings show that signals transmitted through the primary cilium promote the escape of future GABAergic interneurons from their tangential routes to colonize the cortical plate. |
23259945 | NMDA receptor regulation prevents regression of visual cortical function in the absence of Mecp2. | Brain function is shaped by postnatal experience and vulnerable to disruption of Methyl-CpG-binding protein, Mecp2, in multiple neurodevelopmental disorders. How Mecp2 contributes to the experience-dependent refinement of specific cortical circuits and their impairment remains unknown. We analyzed vision in gene-targeted mice and observed an initial normal development in the absence of Mecp2. Visual acuity then rapidly regressed after postnatal day P35-40 and cortical circuits largely fell silent by P55-60. Enhanced inhibitory gating and an excess of parvalbumin-positive, perisomatic input preceded the loss of vision. Both cortical function and inhibitory hyperconnectivity were strikingly rescued independent of Mecp2 by early sensory deprivation or genetic deletion of the excitatory NMDA receptor subunit, NR2A. Thus, vision is a sensitive biomarker of progressive cortical dysfunction and may guide novel, circuit-based therapies for Mecp2 deficiency. |
23259946 | ADF/cofilin-mediated actin retrograde flow directs neurite formation in the developing brain. | Neurites are the characteristic structural element of neurons that will initiate brain connectivity and elaborate information. Early in development, neurons are spherical cells but this symmetry is broken through the initial formation of neurites. This fundamental step is thought to rely on actin and microtubule dynamics. However, it is unclear which aspects of the complex actin behavior control neuritogenesis and which molecular mechanisms are involved. Here, we demonstrate that augmented actin retrograde flow and protrusion dynamics facilitate neurite formation. Our data indicate that a single family of actin regulatory proteins, ADF/Cofilin, provides the required control of actin retrograde flow and dynamics to form neurites. In particular, the F-actin severing activity of ADF/Cofilin organizes space for the protrusion and bundling of microtubules, the backbone of neurites. Our data reveal how ADF/Cofilin organizes the cytoskeleton to drive actin retrograde flow and thus break the spherical shape of neurons. |
23259943 | The role of medial prefrontal cortex in memory and decision making. | Some have claimed that the medial prefrontal cortex (mPFC) mediates decision making. Others suggest mPFC is selectively involved in the retrieval of remote long-term memory. Yet others suggests mPFC supports memory and consolidation on time scales ranging from seconds to days. How can all these roles be reconciled? We propose that the function of the mPFC is to learn associations between context, locations, events, and corresponding adaptive responses, particularly emotional responses. Thus, the ubiquitous involvement of mPFC in both memory and decision making may be due to the fact that almost all such tasks entail the ability to recall the best action or emotional response to specific events in a particular place and time. An interaction between multiple memory systems may explain the changing importance of mPFC to different types of memories over time. In particular, mPFC likely relies on the hippocampus to support rapid learning and memory consolidation. |
23259944 | Microstimulation activates a handful of muscle synergies. | Muscle synergies have been proposed as a mechanism to simplify movement control. Whether these coactivation patterns have any physiological reality within the nervous system remains unknown. Here we applied electrical microstimulation to motor cortical areas of rhesus macaques to evoke hand movements. Movements tended to converge toward particular postures, driven by synchronous bursts of muscle activity. Across stimulation sites, the muscle activations were reducible to linear sums of a few basic patterns-each corresponding to a muscle synergy evident in voluntary reach, grasp, and transport movements made by the animal. These synergies were represented nonuniformly over the cortical surface. We argue that the brain exploits these properties of synergies-postural equivalence, low dimensionality, and topographical representation-to simplify motor planning, even for complex hand movements. |
23259942 | The autism sequencing consortium: large-scale, high-throughput sequencing in autism spectrum disorders. | Research during the past decade has seen significant progress in the understanding of the genetic architecture of autism spectrum disorders (ASDs), with gene discovery accelerating as the characterization of genomic variation has become increasingly comprehensive. At the same time, this research has highlighted ongoing challenges. Here we address the enormous impact of high-throughput sequencing (HTS) on ASD gene discovery, outline a consensus view for leveraging this technology, and describe a large multisite collaboration developed to accomplish these goals. Similar approaches could prove effective for severe neurodevelopmental disorders more broadly. |
23259941 | The need for speed: eye-position signal dynamics in the parietal cortex. | Accurate eye-position signals are critically important for localizing targets in space when the eyes move. In this issue of Neuron, Xu et al. (2012) provide evidence that eye-position gain fields in area LIP remain spatially inaccurate for some time after a saccade, indicating they are not updated rapidly enough to play a role in the computation of target locations for upcoming saccades. |
23259940 | There and back again: the corticobulbar loop. | Feedback is a ubiquitous anatomical feature of sensory processing in vertebrates. In this issue of Neuron, two papers (Boyd et al., 2012, and Markopoulos et al., 2012) analyze the features of feedback from olfactory cortex to olfactory bulb. |
23259939 | Stimulating news about modular motor control. | How does the motor system efficiently control dexterous finger movements? A study by Overduin et al. (2012) shows that muscle activity patterns elicited by cortical microstimulation matched those extracted from natural movements and hence could constitute the building blocks for movement production. |
23259937 | Invasive pneumococcal disease after routine pneumococcal conjugate vaccination in children, England and Wales. | We assessed known risk factors, clinical presentation, and outcome of invasive pneumococcal disease (IPD) in children 3-59 months of age after introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) in England and Wales. During September 2006-March 2010, a total of 1,342 IPD episodes occurred in 1,332 children; 14.9% (198/1,332) had comorbidities. Compared with IPD caused by PCV7 serotypes (44/248; 17.7%), comorbidities were less common for the extra 3 serotypes in the 10-valent vaccine (15/299; 5.0%) but similar to the 3 additional PCV13 serotypes (45/336; 13.4%) and increased for the 11 extra serotypes in 23-valent polysaccharide vaccine (PPV23) (39/186; 21.0%) and non-PPV23 serotypes (38/138; 27.5%). Fifty-two (3.9%) cases resulted from PCV7 failure; 9 (0.7%) case-patients had recurrent IPD. Case-fatality rate was 4.4% (58/1,332) but higher for meningitis (11.0%) and children with comorbidities (9.1%). Thus, comorbidities were more prevalent in children with IPD caused by non-PCV13 serotypes and were associated with increased case fatality. |
23259936 | Systematic investigation of relationship between strength of NH···S hydrogen bond and reactivity of molybdoenzyme models. | A series of monooxomolybdenum(IV) and dioxomolybdenum(VI) complexes containing two intramolecular NH···S hydrogen bonds were synthesized and characterized by (1)H NMR, UV-visible, IR, and Raman spectroscopies, and electrochemical measurements. Elimination of the steric effects enabled the accurate and quantitative evaluation of NH···S hydrogen bonds. Strong correlations among the strength of the hydrogen bond, the strength of the Mo(VI)═O bond, and the redox potential were clearly shown. The hydrogen bonds stabilize the intermediate in the reaction between the monooxomolybdenum(IV) and Me(3)NO, resulting in acceleration of the reaction or retardation of trans-cis rearrangement. The proposed intermediate and the reaction mechanism, discussed on the basis of theoretical calculations, provided a unified explanation of the reaction. |
23259935 | Tailorable cell culture platforms from enzymatically cross-linked multifunctional poly(ethylene glycol)-based hydrogels. | As stem-cell-based therapies rapidly advance toward clinical applications, there is a need for cheap, easily manufactured, injectable gels that can be tailored to carry stem cells and impart function to such cells. Herein we describe a process for making hydrogels composed of hydroxyphenyl propionic acid (HPA) conjugated, branched poly(ethylene glycol) (PEG) via an enzyme mediated, oxidative cross-linking method. Functionalization of the branched PEG with HPA at varying degrees of substitution was confirmed via attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and (1)H NMR. The versatility of this hydrogel system was exemplified through variations in the degree of HPA substitution, polymer concentration, and the concentration of cross-linking reagents (horseradish peroxidase and H(2)O(2)), which resulted in a range of mechanical properties and gelation kinetics for these gels. Cross-linking of the PEG-HPA conjugate with a recombinantly produced Fibronectin fragment (Type III domains 7-10) encouraged attachment and spreading of human mesenchymal stem cells (hMSCs) when assessed in both two-dimensional and three-dimensional formats. Interestingly, when encapsulated in both nonfunctionalized and functionalized cross-linked PEG-HPA gels, MSCs showed good viability over all time periods assessed. With tunable gelation kinetics and mechanical properties, these hydrogels provide a flexible in vitro cell culture platform that will likely have significant utility in tissue engineering as an injectable delivery platform for cells to sites of tissue damage. |
23259934 | Doctors' and nurses' views on patient care for type 2 diabetes: an interview study in primary health care in Oman. | This study aimed at exploring the experiences of primary health-care providers of their encounters with patients with type 2 diabetes, and their preferences and suggestions for future improvement of diabetes care. Barriers to good diabetes care could be related to problems from health-care providers' side, patients' side or the health-care system of the country. Treatment of patients with type 2 diabetes has become a huge challenge in Oman, where the prevalence has increased to high levels. Semi-structured interviews were conducted with 26 health-care professionals, 19 doctors and seven nurses, who worked in primary health care in Oman. Qualitative content analysis was applied. Findings Organizational barriers and barriers related to patients and health-care providers were identified. These included workload and lack of teamwork approach. Poor patients' management adherence and influence of culture on their attitudes towards illness were identified. From the providers' side, language barriers, providers' frustration and aggressive attitudes towards the patients were reflected. Decreasing the workload, availability of competent teams with diabetes specialist nurses and continuity of care were suggested. Furthermore, changing professional behaviours towards a more patient-centred approach and need for health education to the patients, especially on self-management, were addressed. Appropriate training for health-care providers in communication skills with emphasis on self-care education and individualization of care according to each patient's needs are important for improvement of diabetes care in Oman. |
23259933 | Evaluating changes in radiation treatment volumes from post-operative to same-day planning MRI in High-grade gliomas. | Adjuvant radiation therapy (RT) with temozolomide (TMZ) is standard of care for high grade gliomas (HGG) patients. RT is commonly started 3 to 5 weeks after surgery. The deformation of the tumor bed and brain from surgery to RT is poorly studied. This study examined the magnitude of volume change in the postoperative tumor bed and the potential impact of RT planning. This study includes 24 patients with HGG who underwent craniotomy and adjuvant RT with TMZ at our institution. All patients had immediate postoperative MRI and repeat MRI during the day of RT simulation. Gross tumor volumes (GTV), clinical target volumes (CTV) of initial 46 Gy (CTV1) and boost to 60 Gy (CTV2) were contoured on both sets of MRIs according to RTOG (Radiation Therapy Oncology Group) guidelines. For patients who recurred after RT, the recurrence pattern was evaluated. An average of 17 days elapsed between immediate and delayed MRIs. GTV1 (FLAIR abnormality and tumor bed) decreased significantly on the delayed MRI as compared to immediate post-operative MRI (mean = 30.96cc, p = 0.0005), while GTV2 (contrast-enhanced T1 abnormality and tumor bed) underwent a non-significant increase (mean = 6.82cc, p = 0.07). Such changes lead to significant decrease of CTV1 (mean decrease is 113.9cc, p<0.01), and significant increase of CTV2 (mean increase is 32.5cc, p=0.05). At a median follow-up of 13 months, 16 patients (67%) progressed, recurred, or died, with a progression-free survival time of 13.7 months. Twelve patients failed within all CTVs based on immediate and delayed MRIs, while one patient recurred outside of CTV2 based on immediate post-operative MRI, but within the CTV2 defined on delayed MRI. The postoperative tumor bed of HGGs undergoes substantial volumetric changes after surgery. Treatment planning based on delayed MRI significantly reduces the volume of treated brain tissue without local control detriment. The marked reduction of volume treated to 46 Gy based on delayed MRI scan, could result in increased sparing of organs at risk. There may be a small risk of inadequate radiation field design if radiation planning is based on immediate post-operative MRI. |
23259930 | Lack of evidence to support the association of a single IL28B genotype SNP rs12979860 with the HTLV-1 clinical outcomes and proviral load. | The Interleukin 28B (IL28B) rs12979860 polymorphisms was recently reported to be associated with the human T-cell leukemia virus type 1 (HTLV-1) proviral load (PvL) and the development of the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). In an attempt to examine this hypothesis, we assessed the association of the rs12979860 genotypes with HTLV-1 PvL levels and clinical status in 112 unrelated Brazilian subjects (81 HTLV-1 asymptomatic carriers, 24 individuals with HAM/TSP and 7 with Adult T cell Leukemia/Lymphoma (ATLL)). All 112 samples were successfully genotyped and their PvLs compared. Neither the homozygote TT nor the heterozygote CT mutations nor the combination genotypes (TT/CT) were associated with a greater PvL. We also observed no significant difference in allele distribution between asymptomatic carriers and patients with HTLV-1 associated HAM/TSP. Our study failed to support the previously reported positive association between the IL28B rs12979860 polymorphisms and an increased risk of developing HAM/TSP in the Brazilian population. |
23259931 | Perampanel, a novel, non-competitive, selective AMPA receptor antagonist as adjunctive therapy for treatment-resistant partial-onset seizures. | In the search for new antiepileptic drugs (AEDs), AMPA-type receptor antagonists have a novel target and the potential to improve seizure control in patients with refractory seizures. This article reviews preclinical and clinical data for 2-(2-oxo-1-phenyl-5-pyridin-2-yl-1,2-dihydropyridin-3-yl)benzonitrile, perampanel , a new chemical entity developed for the treatment of partial-onset seizures. Perampanel is a selective, non-competitive AMPA receptor antagonist. The preclinical profile of perampanel and its clinical development are reviewed. Unlike many traditional AEDs, perampanel demonstrated efficacy in a broad spectrum of preclinical seizure models. Phase I and II clinical studies suggested perampanel had a favorable safety and tolerability profile and demonstrated proof of concept for its mechanism of action in patients with treatment-resistant partial-onset seizures. Three Phase III studies have additionally demonstrated that adjunctive perampanel 4 - 12 mg/day is well-tolerated and significantly improves seizure control in these patients. Median reductions in seizure frequency were 23.3% (4 mg), 26.3 - 30.8% (8 mg) and 17.6 - 34.5% (12 mg) versus 9.7 - 21.0% for placebo. Responder rates were 28.5% (4 mg), 33.3 - 37.6% (8 mg) and 33.9 - 36.1% (12 mg) versus 14.7 - 26.4% for placebo. Perampanel may offer an alternative treatment option in the management of patients with refractory partial-onset seizures. |
23259929 | The effect of glial fibrillary acidic protein expression on neurite outgrowth from retinal explants in a permissive environment. | Increased expression of glial fibrillary acidic protein (GFAP) within macroglia is commonly seen as a hallmark of glial activation after damage within the central nervous system, including the retina. The increased expression of GFAP in glia is also considered part of the pathologically inhibitory environment for regeneration of axons from damaged neurons. Recent studies have raised the possibility that reactive gliosis and increased GFAP cannot automatically be assumed to be negative events for the surrounding neurons and that the context of the reactive gliosis is critical to whether neurons benefit or suffer. We utilized transgenic mice expressing a range of Gfap to titrate the amount of GFAP in retinal explants to investigate the relationship between GFAP concentration and the regenerative potential of retinal ganglion cells. Explants from Gfap-/- and Gfap+/- mice did not have increased neurite outgrowth compared with Gfap+/+ or Gfap over-expressing mice as would be expected if GFAP was detrimental to axon regeneration. In fact, Gfap over-expressing explants had the most neurite outgrowth when treated with a neurite stimulatory media. Transmission electron microscopy revealed that neurites formed bundles, which were surrounded by larger cellular processes that were GFAP positive indicating a close association between growing axons and glial cells in this regeneration paradigm. We postulate that glial cells with increased Gfap expression support the elongation of new neurites from retinal ganglion cells possibly by providing a scaffold for outgrowth. |
23259928 | Notch signaling imparts and preserves neural stem characteristics in the adult brain. | Although the role of Notch has been studied extensively in the developing nervous system, the embryonic lethality of Notch pathway mutants has hindered studies in the adult brain. The creation of cre/lox-mediated conditional gain- and loss-of-function mice has allowed us to investigate the role of Notch signaling in adult neural stem and progenitor cells. We have determined that Notch signaling is important for conferring stem cell characteristics upon neural precursor cells. Knocking-out Notch signaling in vivo results in neural progenitors, leaving the subependymal niche and migrating along the rostral migratory stream to the olfactory bulb, while overexpressing Notch results in retention of cells in the subependyma. Further, increased Notch signaling in progenitor cells resulted in the expression of stem cell markers in vivo as well as conferring the characteristics of self-renewal and multipotentiality upon subsequent isolation in vitro. Similar to what has been reported from the embryonic brain, the overexpression of Notch in neural precursor cells in vitro increased the numbers of neurospheres from the adult brain. Finally, overexpression of Notch1 in pure populations of progenitor cells (excluding neural stem cells) isolated by fluorescence activated cell sorting led to the formation of multipotent, self-renewing neurospheres from the non-neurosphere forming fraction. Hence, Notch overexpression confers stem cell properties upon progenitor cells and demonstrates that Notch signaling not only preserves stem cell characteristics, but that it can confer stem cell characteristics upon a subset of progenitor cells. |
23259927 | Two-dimensional ultrathin gold film composed of steadily linked dense nanoparticle with surface plasmon resonance. | Noble metallic nanoparticles have prominent optical local-field enhancement and light trapping properties in the visible light region resulting from surface plasmon resonances. We investigate the optical spectral properties and the surface-enhanced Raman spectroscopy of two-dimensional distinctive continuous ultrathin gold nanofilms. Experimental results show that the one- or two-layer nanofilm obviously increases absorbance in PEDOT:PSS and P3HT:PCBM layers and the gold nanofilm acquires high Raman-enhancing capability. The fabricated novel structure of the continuous ultrathin gold nanofilms possesses high surface plasmon resonance properties and boasts a high surface-enhanced Raman scattering (SERS) enhancement factor, which can be a robust and cost-efficient SERS substrate. Interestingly, owing to the distinctive morphology and high light transmittance, the peculiar nanofilm can be used in multilayer photovoltaic devices to trap light without affecting the physical thickness of solar photovoltaic absorber layers and yielding new options for solar cell design. |
23259926 | Influence of birth order, birth weight, colostrum and serum immunoglobulin G on neonatal piglet survival. | Intake of colostrum after birth is essential to stimulate intestinal growth and function, and to provide systemic immunological protection via absorption of Immunoglobulin G (IgG). The birth order and weight of 745 piglets (from 75 litters) were recorded during a one-week period of farrowing. Only pigs weighing greater than 0.68 kg birth weight were chosen for the trial. Sow colostrum was collected during parturition, and piglets were bled between 48 and 72 hours post-birth. Piglet serum IgG and colostral IgG concentrations were determined by radial immunodiffusion. Sow parity had a significant (P < 0.001) effect on sow colostral IgG concentration, being 5% higher in multiparous females. Sow colostral IgG concentration explained 6% and piglet birth order accounted for another 4% of the variation observed in piglet serum IgG concentration (P < 0.05); however, birth weight had no detectable effect. Piglet serum IgG concentration had both a linear (P < 0.05) and quadratic effect (P < 0.05) on % survival. Piglets with 1,000 mg/dl serum IgG or less (n=24) had a 67% survival; whereas, piglets with IgG concentrations between 2250 to 2500 mg/dl (n=247) had a 91% survival. Birth order had no detectable effect on survival, but birth weight had a positive linear effect (P < 0.05). Piglets weighing 0.9 kg (n = 107) at birth had a 68% survival rate, and those weighing 1.6 kg (n = 158) had an 89% survival. We found that the combination of sow colostrum IgG concentration and birth order can account for 10% of the variation of piglet serum IgG concentration and that piglets with less than 1,000 mg/dl IgG serum concentration and weight of 0.9 kg at birth had low survival rate when compared to their larger siblings. The effective management of colostrum uptake in neonatal piglets in the first 24 hrs post-birth may potentially improve survival from birth to weaning. |
23259924 | Moderate prevalence of transmitted drug resistance mutations among antiretroviral-naive HIV-infected pregnant women in Rio de Janeiro, Brazil. | Transmission of drug-resistant HIV-1 strains has been gaining attention and is becoming a growing problem throughout the world. The aim of this study was to determine the prevalence of transmitted drug resistance mutations (TDRM) among antiretroviral (ARV)-naive HIV-infected pregnant women in Rio de Janeiro, Brazil. ARV-naive pregnant women were recruited at Hospital Geral de Nova Iguacu (HGNI), Rio de Janeiro, from 2005 to 2008. HIV genotyping was carried out using ViroSeq (Abbott v. 2.0). TDRM were detected using the Calibrated Population Resistance Tool-CPR v. 6.0.The prevalence of mutations associated with resistance in the protease and reverse transcriptase regions of the HIV genome were assessed in samples collected prior to initiation of ARV prophylaxis or treatment. Among 238 eligible specimens that were collected, 197 samples were successfully amplified using reverse transcription polymerase chain reaction. Eighty-one percent of women were infected with HIV subtype B, 10% with subtype F1 viruses, 1.0% with subtype C virus, and 8.0% with recombinant forms of the virus. The prevalence of HIV TDRM was 5.6% for nucleoside reverse transcriptase inhibitors, 2.0% for nonnucleoside reverse transcriptase inhibitors, and 3.0% for protease inhibitors. The overall prevalence of any drug resistance was 10.7%. There were no multiclass resistant strains identified in the analyzed samples. The prevalence of HIV TDRM among the pregnant women in our cohort was moderate. Resistance testing should be encouraged in Rio de Janeiro, among other locations, for all HIV-infected pregnant women prior to prevention of mother-to-child transmission of HIV. |