The molecular clockwork of the fire ant Solenopsis invicta.
The circadian clock is a core molecular mechanism that allows organisms to anticipate daily environmental changes and adapt the timing of behaviors to maximize efficiency. In social insects, the ability to maintain the appropriate temporal order is thought to improve colony efficiency and fitness. We used the newly sequenced fire ant (Solenopsis invicta) genome to characterize the first ant circadian clock. Our results reveal that the fire ant clock is similar to the clock of the honeybee, a social insect with an independent evolutionary origin of sociality. Gene trees for the eight core clock genes, period, cycle, clock, cryptochrome-m, timeout, vrille, par domain protein 1 & clockwork orange, show ant species grouping closely with honeybees and Nasonia wasps as an outgroup to the social Hymenoptera. Expression patterns for these genes suggest that the ant clock functions similar to the honeybee clock, with period and cry-m mRNA levels increasing during the night and cycle and clockwork orange mRNAs cycling approximately anti-phase to period. Gene models for five of these genes also parallel honeybee models. In particular, the single ant cryptochrome is an ortholog of the mammalian-type (cry-m), rather than Drosophila-like protein (cry-d). Additionally, we find a conserved VPIFAL C-tail region in clockwork orange shared by insects but absent in vertebrates. Overall, our characterization of the ant clock demonstrates that two social insect lineages, ants and bees, share a similar, mammalian-like circadian clock. This study represents the first characterization of clock genes in an ant and is a key step towards understanding socially-regulated plasticity in circadian rhythms by facilitating comparative studies on the organization of circadian clockwork.


Cytosolic triacylglycerol biosynthetic pathway in oilseeds. Molecular cloning and expression of peanut cytosolic diacylglycerol acyltransferase.
Triacylglycerols (TAGs) are the most important storage form of energy for eukaryotic cells. TAG biosynthetic activity was identified in the cytosolic fraction of developing peanut (Arachis hypogaea) cotyledons. This activity was NaF insensitive and acyl-coenzyme A (CoA) dependent. Acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyzes the final step in TAG biosynthesis that acylates diacylglycerol to TAG. Soluble DGAT was identified from immature peanuts and purified by conventional column chromatographic procedures. The enzyme has a molecular mass of 41 +/- 1.0 kD. Based on the partial peptide sequence, a degenerate probe was used to obtain the full-length cDNA. The isolated gene shared less than 10% identity with the previously identified DGAT1 and 2 families, but has 13% identity with the bacterial bifunctional wax ester/DGAT. To differentiate the unrelated families, we designate the peanut gene as AhDGAT. Expression of peanut cDNA in Escherichia coli resulted in the formation of labeled TAG and wax ester from [14C]acetate. The recombinant E. coli showed high levels of DGAT activity but no wax ester synthase activity. TAGs were localized in transformed cells with Nile blue A and oil red O staining. The recombinant and native DGAT was specific for 1,2-diacylglycerol and did not utilize hexadecanol, glycerol-3-phosphate, monoacylglycerol, lysophosphatidic acid, and lysophosphatidylcholine. Oleoyl-CoA was the preferred acyl donor as compared to palmitoyl- and stearoyl-CoAs. These data suggest that the cytosol is one of the sites for TAG biosynthesis in oilseeds. The identified pathway may present opportunities of bioengineering oil-yielding plants for increased oil production.


North Atlantic right whales (Eubalaena glacialis) ignore ships but respond to alerting stimuli.
North Atlantic right whales were extensively hunted during the whaling era and have not recovered. One of the primary factors inhibiting their recovery is anthropogenic mortality caused by ship strikes. To assess risk factors involved in ship strikes, we used a multi-sensor acoustic recording tag to measure the responses of whales to passing ships and experimentally tested their responses to controlled sound exposures, which included recordings of ship noise, the social sounds of conspecifics and a signal designed to alert the whales. The whales reacted strongly to the alert signal, they reacted mildly to the social sounds of conspecifics, but they showed no such responses to the sounds of approaching vessels as well as actual vessels. Whales responded to the alert by swimming strongly to the surface, a response likely to increase rather than decrease the risk of collision.


Ecological modelling in an evolutionary context.
I argue that one of the strong features in disciplines like molecular biology and cosmology is the extent ot which they use a powerful theoretical framework to generate and test quantitative predictions. Studies of biological evolution can exploit a similar advantage by integrating our current understanding of physiological and sociobiological processes to generate models of much greater sophistication than has commonly been the practice hitherto. I illustrate this with a number of examples drawn from the evolutionary biology of human and nonhuman primates.


Intrinsic Folding Properties of the HLA-B27 Heavy Chain Revealed by Single Chain Trimer Versions of Peptide-Loaded Class I Major Histocompatibility Complex Molecules.
Peptide-loaded Major Histocompatibility Complex (pMHC) class I molecules can be expressed in a single chain trimeric (SCT) format, composed of a specific peptide fused to the light chain beta-2 microglobulin (β2m) and MHC class I heavy chain (HC) by flexible linker peptides. pMHC SCTs have been used as effective molecular tools to investigate cellular immunity and represent a promising vaccine platform technology, due to their intracellular folding and assembly which is apparently independent of host cell folding pathways and chaperones. However, certain MHC class I HC molecules, such as the Human Leukocyte Antigen B27 (HLA-B27) allele, present a challenge due to their tendency to form HC aggregates. We constructed a series of single chain trimeric molecules to determine the behaviour of the HLA-B27 HC in a scenario that usually allows for efficient MHC class I molecule folding. When stably expressed, a pMHC SCT incorporating HLA-B27 HC formed chaperone-bound homodimers within the endoplasmic reticulum (ER). A series of HLA-B27 SCT substitution mutations revealed that the F pocket and antigen binding groove regions of the HLA-B27 HC defined the folding and dimerisation of the single chain complex, independently of the peptide sequence. Furthermore, pMHC SCTs can demonstrate variability in their association with the intracellular antigen processing machinery.


Liver-primed memory T cells generated under noninflammatory conditions provide anti-infectious immunity.
Development of CD8(+) T cell (CTL) immunity or tolerance is linked to the conditions during T cell priming. Dendritic cells (DCs) matured during inflammation generate effector/memory T cells, whereas immature DCs cause T cell deletion/anergy. We identify a third outcome of T cell priming in absence of inflammation enabled by cross-presenting liver sinusoidal endothelial cells. Such priming generated memory T cells that were spared from deletion by immature DCs. Similar to central memory T cells, liver-primed T cells differentiated into effector CTLs upon antigen re-encounter on matured DCs even after prolonged absence of antigen. Their reactivation required combinatorial signaling through the TCR, CD28, and IL-12R and controlled bacterial and viral infections. Gene expression profiling identified liver-primed T cells as a distinct Neuropilin-1(+) memory population. Generation of liver-primed memory T cells may prevent pathogens that avoid DC maturation by innate immune escape from also escaping adaptive immunity through attrition of the T cell repertoire.


Multiple Minor QTLs Are Responsible for Fusarium Head Blight Resistance in Chinese Wheat Landrace Haiyanzhong.
Fusarium head blight (FHB), caused by Fusarium graminearum Schwabe, is a devastating disease in wheat (Triticum aestivum L.). Use of host resistance is one of the most effective strategies to minimize the disease damage. Haiyanzhong (HYZ) is a Chinese wheat landrace that shows a high level of resistance to FHB spread within a spike (type II resistance). To map the quantitative trait loci (QTLs) in HYZ and identify markers tightly linked to the QTLs for FHB resistance, a population of 172 recombinant inbred lines (RILs) from a cross between HYZ and Wheaton (FHB susceptible) was genotyped using simple sequence repeats (SSRs) and single-nucleotide polymorphisms (SNPs) derived from genotyping-by-sequencing (GBS), and evaluated for percentage of symptomatic spikelets (PSSs) per spike in three greenhouse experiments. Six QTLs for type II resistance were identified in HYZ, indicating that multiple minor QTLs together can provide a high level of FHB resistance in wheat. The QTL with the largest effect on FHB resistance was mapped on the chromosome arm 5AS, and the other five from HYZ were mapped on the chromosomes 6B, 7D, 3B, 4B and 4D. In addition, two QTLs from Wheaton were mapped on 2B. Critical SNPs linked to the QTLs on chromosomes 5A, 6B, and 2B were converted into KBioscience competitive allele-specific PCR (KASP) assays, which can be used for marker-assisted selection (MAS) to pyramid these QTLs in wheat.


Targeting the RAS pathway by mitogen-activated protein kinase inhibitors.
Targeting of oncogenic driver mutations with small-molecule inhibitors resulted in powerful treatment options for cancer patients in recent years. The RAS (rat sarcoma) pathway is among the most frequently mutated pathways in human cancer. Whereas targeting mutant Kirsten RAS (KRAS) remains difficult, mutant B rapidly accelerated fibrosarcoma (BRAF) kinase is an established drug target in cancer. Now data show that neuroblastoma RAS (NRAS) and even Harvey RAS (HRAS) mutations could be predictive markers for treatment with mitogen-activated protein kinase (MEK) inhibitors. This review discusses recent preclinical and clinical studies of MEK inhibitors in BRAF and RAS mutant cancer.


Early Blood Pressure Reduction in Acute Ischemic Stroke with Various Severities: A Subgroup Analysis of the CATIS Trial.
Clinical trials have generally showed a neutral effect of blood pressure (BP) reduction on clinical outcomes among acute ischemic stroke patients. We conducted a prespecified subgroup analysis to assess whether disease severity modifies the effect of early antihypertensive treatment on death and disability among patients with acute ischemic stroke. In the China Antihypertensive Trial in Acute Ischemic Stroke, 4,071 patients with acute ischemic stroke and elevated BP were randomly assigned to receive antihypertensive treatment or to discontinue all hypertension medications within 48 h of symptom onset. The primary outcome was a combination of death and major disability at 14 days or hospital discharge. In this subgroup analysis, participants were categorized into 3 groups according to their baseline NIH Stroke Scale (NIHSS) scores (0-4, 5-15, or ≥16). At 24 h after randomization, mean systolic BP differences (95% CIs) were -8.5 (-10.0 to -7.1), -9.8 (-11.4 to -8.3), and -9.1 (-14.4 to -3.8) mm Hg between the treatment and control groups (all p values <0.001) for patients with a baseline NIHSS score of 0-4, 5-15, and ≥16, respectively. At day 7 after randomization, the corresponding mean systolic BP differences were -9.3 (-10.5 to -8.2), -9.1 (-10.3 to -7.8), and -10.1 (-15.1 to -5.1) mm Hg between the treatment and control groups (all p values <0.001). The primary outcome was not significantly different between the treatment and control groups at day 14 or hospital discharge among all NIHSS subgroups (p value for homogeneity = 0.66). ORs (95% CI) associated with treatment were 1.14 (0.87-1.49, p = 0.33), 1.04 (0.86-1.25, p = 0.70), and 0.67 (0.18-2.44, p = 0.54) for patients with a baseline NIHSS score of 0-4, 5-15, and ≥16, respectively. The composite outcome of death and major disability at 3-month follow-up did not differ between the 2 comparison groups for all NIHSS subgroups. In addition, vascular events and recurrent stroke were not significantly different between the 2 comparison groups at the 3-month follow-up visit among all NIHSS subgroups except that there was a suggestive risk reduction for recurrent stroke among those with an NIHSS score of 5-15 (OR 0.45, 95% CI 0.20-0.99, p = 0.05). Early BP reduction with antihypertensive medications did not reduce or increase the risk of death, major disabilities, recurrent instances of stroke, and vascular events in acute ischemic stroke patients with a variety of disease severities.


Fluorescence techniques: shedding light on ligand-receptor interactions.
The ability of organisms, or individual cells, to react to external chemical signals, which are detected and transduced by cell-surface receptors, is crucial for their survival. These receptors are the targets of the majority of clinically used medicines. Combinatorial genetics can provide almost unlimited numbers of mutant receptor proteins and combinatorial chemistry can produce large libraries of potential therapeutic compounds that act on these membrane receptors. What is missing for the fundamental understanding of receptor function and for the discovery of new medicines are efficient procedures to screen both ligand-receptor interactions and the subsequent functional consequences. Ultrasensitive fluorescence spectroscopic approaches, in combination with efficient labelling protocols, offer enormous possibilities for highly parallel functional bioanalytics at the micro- and nanometer level.


Tissue Transglutaminase contributes to experimental multiple sclerosis pathogenesis and clinical outcome by promoting macrophage migration.
Multiple sclerosis is a serious neurological disorder, resulting in e.g., sensory, motor and cognitive deficits. A critical pathological aspect of multiple sclerosis (MS) is the influx of immunomodulatory cells into the central nervous system (CNS). Identification of key players that regulate cellular trafficking into the CNS may lead to the development of more selective treatment to halt this process. The multifunctional enzyme tissue Transglutaminase (TG2) can participate in various inflammation-related processes, and is known to be expressed in the CNS. In the present study, we question whether TG2 activity contributes to the pathogenesis of experimental MS, and could be a novel therapeutic target. In human post-mortem material, we showed the appearance of TG2 immunoreactivity in leukocytes in MS lesions, and particular in macrophages in rat chronic-relapsing experimental autoimmune encephalomyelitis (cr-EAE), an experimental MS model. Clinical deficits as observed in mouse EAE were reduced in TG2 knock-out mice compared to littermate wild-type mice, supporting a role of TG2 in EAE pathogenesis. To establish if the enzyme TG2 represents an attractive therapeutic target, cr-EAE rats were treated with TG2 activity inhibitors during ongoing disease. Reduction of TG2 activity in cr-EAE animals dramatically attenuated clinical deficits and demyelination. The mechanism underlying these beneficial effects pointed toward a reduction in macrophage migration into the CNS due to attenuated cytoskeletal flexibility and RhoA GTPase activity. Moreover, iNOS and TNFα levels were selectively reduced in the CNS of cr-EAE rats treated with a TG2 activity inhibitor, whereas other relevant inflammatory mediators were not affected in CNS or spleen by reducing TG2 activity. We conclude that modulating TG2 activity opens new avenues for therapeutic intervention in MS which does not affect peripheral levels of inflammatory mediators.


Synthetic control of mammalian-cell motility by engineering chemotaxis to an orthogonal bioinert chemical signal.
Directed migration of diverse cell types plays a critical role in biological processes ranging from development and morphogenesis to immune response, wound healing, and regeneration. However, techniques to direct, manipulate, and study cell migration in vitro and in vivo in a specific and facile manner are currently limited. We conceived of a strategy to achieve direct control over cell migration to arbitrary user-defined locations, independent of native chemotaxis receptors. Here, we show that genetic modification of cells with an engineered G protein-coupled receptor allows us to redirect their migration to a bioinert drug-like small molecule, clozapine-N-oxide (CNO). The engineered receptor and small-molecule ligand form an orthogonal pair: The receptor does not respond to native ligands, and the inert drug does not bind to native cells. CNO-responsive migration can be engineered into a variety of cell types, including neutrophils, T lymphocytes, keratinocytes, and endothelial cells. The engineered cells migrate up a gradient of the drug CNO and transmigrate through endothelial monolayers. Finally, we demonstrate that T lymphocytes modified with the engineered receptor can specifically migrate in vivo to CNO-releasing beads implanted in a live mouse. This technology provides a generalizable genetic tool to systematically perturb and control cell migration both in vitro and in vivo. In the future, this type of migration control could be a valuable module for engineering therapeutic cellular devices.


The effects of platelet-derived growth factor-BB on periodontal cells in an in vitro wound model.
Platelet-derived growth factor (PDGF-BB) has been shown to enhance periodontal regeneration. Principles of guided tissue regeneration dictate that one of the goals of therapy is to modulate the wound healing processes to favor repopulation of the wound with cells derived from the periodontal ligament rather than from the gingival tissues. Using an in vitro wound model, gingival fibroblasts (GF) have been shown to fill a wound space significantly faster than periodontal ligament cells (PDL). There are no data reported directly comparing the response of these 2 cell types to PDGF-BB within such a wound model. Therefore, the aims of this research were: 1) to characterize both the proliferative and wound fill (WF) effects of PDGF-BB within an in vitro model and 2) to compare specific growth factor effects between GF and PDL. Primary cultures of both human PDL and GF were derived from explanted tissues and passaged to 12-well tissue culture plates. Triplicate cultures of both cell types were grown to confluence and in vitro wounds were mechanically created, removing a 3 mm wide band of the cell layer across the diameter of the wells. The wells were then incubated for 2, 6, and 9 days in media containing 0.1% fetal bovine serum (FBS) and 1 of 5 concentrations of PDGF-BB. At each time point, cells were pulsed with 5-bromo, 2-deoxyuridine (BrdU) fixed, and nuclei were stained to measure BrdU incorporation (as a measure for proliferation). Cells were counter-stained with cytoplasmic stain to measure cell number. Quantitative analyses within the wound boundaries, marginally (area of interest [AOI] 1) and centrally (AOI 2), were accomplished using computer-assisted histomorphometry. PDL exhibited a significantly greater proliferative response to PDGF-BB in both AOI when compared to GF (P <0.0001). The PDL exhibited increased levels of proliferation at concentrations of PDGF-BB greater than or equal to 10 ng/ml. By contrast, GF displayed no increase in proliferation in response to stimulation with PDGF-BB at any of the concentrations tested when compared to negative controls. The wound fill (WF) responses to PDGF-BB were similar between PDL and GF, with both cell types responding in an all or none fashion when measured at day 2, and in a concentration-dependent manner at later time points. The only significant difference in WF between PDL and GF occurred in AOI 2 in negative control medium (0 ng/ml of PDGF-BB), with GFs having greater (P <0.01) levels of WF over the 9 days. The findings from this study demonstrate differing effects of PDGF-BB on the proliferation of PDL and GF in this in vitro model. These results suggest that there may be cell-specific differences critical to periodontal wound healing that may be exploited in the development of new therapies.


Research into the chronodisruption-cancer theory: the imperative for causal clarification and the danger of causal reductionism.
A cascade of investigations into the validity of the chronodisruption-cancer theory (CD-CT), proposed 2008 in Neuroendocrinology Letters as a synthesis of abundant experimental and tantalizing - but still limited - epidemiological data, can be expected in coming years. In view of the possible public health relevance of the suggested links between chronodisruption, defined as the disruption of the temporal organization or order of biological rhythmicity over days and seasons, and the development of disease, including cancer, we provide practical aspects which should be considered for the planning, conduct and the very interpretation of such studies. The interrelated considerations address two facets: first, the critical necessity of causal clarification can be evinced appropriately by Poole's earlier contribution to the Journal. Researchers into the validity of the CD-CT should certainly consider Poole's 2002 illustration via causal graphs for shift work, light at night and breast cancer of the subtleties that can arise in the use of exposure surrogates of different kinds. Second, we emphasize the importance of considering the Zeitgeber multiplicity to avoid causal reductionism in epidemiologic - but also other - chronodisruption research.


Artificial neural network classification of pharyngeal high-resolution manometry with impedance data.
To use classification algorithms to classify swallows as safe, penetration, or aspiration based on measurements obtained from pharyngeal high-resolution manometry (HRM) with impedance. Case series evaluating new method of data analysis. Multilayer perceptron, an artificial neural network (ANN), was evaluated for its ability to classify swallows as safe, penetration, or aspiration. Data were collected from 25 disordered subjects swallowing 5- or 10-mL boluses. Following extraction of relevant parameters, a subset of the data was used to train the models, and the remaining swallows were then independently classified by the ANN. A classification accuracy of 89.4 ± 2.4% was achieved when including all parameters. Including only manometry-related parameters yielded a classification accuracy of 85.0 ± 6.0%, whereas including only impedance-related parameters yielded a classification accuracy of 76.0 ± 4.9%. Receiver operating characteristic analysis yielded areas under the curve of 0.8912 for safe, 0.8187 for aspiration, and 0.8014 for penetration. Classification models show high accuracy in classifying swallows from dysphagic patients as safe or unsafe. HRM-impedance with ANN represents one method that could be used clinically to screen for patients at risk for penetration or aspiration.


Computational approaches to understand cardiac electrophysiology and arrhythmias.
Cardiac rhythms arise from electrical activity generated by precisely timed opening and closing of ion channels in individual cardiac myocytes. These impulses spread throughout the cardiac muscle to manifest as electrical waves in the whole heart. Regularity of electrical waves is critically important since they signal the heart muscle to contract, driving the primary function of the heart to act as a pump and deliver blood to the brain and vital organs. When electrical activity goes awry during a cardiac arrhythmia, the pump does not function, the brain does not receive oxygenated blood, and death ensues. For more than 50 years, mathematically based models of cardiac electrical activity have been used to improve understanding of basic mechanisms of normal and abnormal cardiac electrical function. Computer-based modeling approaches to understand cardiac activity are uniquely helpful because they allow for distillation of complex emergent behaviors into the key contributing components underlying them. Here we review the latest advances and novel concepts in the field as they relate to understanding the complex interplay between electrical, mechanical, structural, and genetic mechanisms during arrhythmia development at the level of ion channels, cells, and tissues. We also discuss the latest computational approaches to guiding arrhythmia therapy.


A critical review of the postulated role of the non-essential amino acid, β-N-methylamino-L-alanine, in neurodegenerative disease in humans.
The compound BMAA (β-N-methylamino-L-alanine) has been postulated to play a significant role in four serious neurological human diseases: Amyotrophic Lateral Sclerosis/Parkinsonism Dementia Complex (ALS/PDC) found on Guam, and ALS, Parkinsonism, and dementia that occur globally. ALS/PDC with symptoms of all three diseases first came to the attention of the scientific community during and after World War II. It was initially associated with cycad flour used for food because BMAA is a product of symbiotic cycad root-dwelling cyanobacteria. Human consumption of flying foxes that fed on cycad seeds was later suggested as a source of BMAA on Guam and a cause of ALS/PDC. Subsequently, the hypothesis was expanded to include a causative role for BMAA in other neurodegenerative diseases including Alzheimer's disease (AD) through exposures attributed to proximity to freshwaters and/or consumption of seafood due to its purported production by most species of cyanobacteria. The hypothesis that BMAA is the critical factor in the genesis of these neurodegenerative diseases received considerable attention in the medical, scientific, and public arenas. This review examines the history of ALS/PDC and the BMAA-human disease hypotheses; similarities and differences between ALS/PDC and the other diseases with similar symptomologies; the relationship of ALS/PDC to other similar diseases, studies of BMAA-mediated effects in lab animals, inconsistencies and data gaps in the hypothesis; and other compounds and agents that were suggested as the cause of ALS/PDC on Guam. The review concludes that the hypothesis of a causal BMAA neurodegenerative disease relationship is not supported by existing data.


Hormesis does not make sense except in the light of TOR-driven aging.
Weak stresses (including weak oxidative stress, cytostatic agents, heat shock, hypoxia, calorie restriction) may extend lifespan. Known as hormesis, this is the most controversial notion in gerontology. For one, it is believed that aging is caused by accumulation of molecular damage. If so, hormetic stresses (by causing damage) must shorten lifespan. To solve the paradox, it was suggested that, by activating repair, hormetic stresses eventually decrease damage. Similarly, Baron Munchausen escaped from a swamp by pulling himself up by his own hair. Instead, I discuss that aging is not caused by accumulation of molecular damage. Although molecular damage accumulates, organisms do not live long enough to age from this accumulation. Instead, aging is driven by overactivated signal-transduction pathways including the TOR (Target of Rapamycin) pathway. A diverse group of hormetic conditions can be divided into two groups. "Hormesis A" inhibits the TOR pathway. "Hormesis B" increases aging-tolerance, defined as the ability to survive catastrophic complications of aging. Hormesis A includes calorie restriction, resveratrol, rapamycin, p53-inducing agents and, in part, physical exercise, heat shock and hypoxia. Hormesis B includes ischemic preconditioning and, in part, physical exercise, heat shock, hypoxia and medical interventions.


3D printing of silk fibroin-based hybrid scaffold treated with platelet rich plasma for bone tissue engineering.
3D printing/bioprinting are promising techniques to fabricate scaffolds with well controlled and patient-specific structures and architectures for bone tissue engineering. In this study, we developed a composite bioink consisting of silk fibroin (SF), gelatin (GEL), hyaluronic acid (HA), and tricalcium phosphate (TCP) and 3D bioprinted the silk fibroin-based hybrid scaffolds. The 3D bioprinted scaffolds with dual crosslinking were further treated with human platelet-rich plasma (PRP) to generate PRP coated scaffolds. Live/Dead and MTT assays demonstrated that PRP treatment could obviously promote the cell growth and proliferation of human adipose derived mesenchymal stem cells (HADMSC). In addition, the treatment of PRP did not significantly affect alkaline phosphatase (ALP) activity and expression, but significantly upregulated the gene expression levels of late osteogenic markers. This study demonstrated that the 3D printing of silk fibroin-based hybrid scaffolds, in combination with PRP post-treatment, might be a more efficient strategy to promote osteogenic differentiation of adult stem cells and has significant potential to be used for bone tissue engineering.


Using reduced catalysts for oxidation reactions: mechanistic studies of the "Periana-Catalytica" system for CH4 oxidation.
Designing oxidation catalysts based on CH activation with reduced, low oxidation state species is a seeming dilemma given the proclivity for catalyst deactivation by overoxidation. This dilemma has been recognized in the Shilov system where reduced Pt(II) is used to catalyze methane functionalization. Thus, it is generally accepted that key to replacing Pt(IV) in that system with more practical oxidants is ensuring that the oxidant does not over-oxidize the reduced Pt(II) species. The "Periana-Catalytica" system, which utilizes (bpym)Pt(II)Cl2 in concentrated sulfuric acid solvent at 200 °C, is a highly stable catalyst for the selective, high yield oxy-functionalization of methane. In lieu of the over-oxidation dilemma, the high stability and observed rapid oxidation of (bpym)Pt(II)Cl2 to Pt(IV) in the absence of methane would seem to contradict the originally proposed mechanism involving CH activation by a reduced Pt(II) species. Mechanistic studies show that the originally proposed mechanism is incomplete and that while CH activation does proceed with Pt(II) there is a solution to the over-oxidation dilemma. Importantly, contrary to the accepted view to minimize Pt(II) overoxidation, these studies also show that increasing that rate could increase the rate of catalysis and catalyst stability. The mechanistic basis for this counterintuitive prediction could help to guide the design of new catalysts for alkane oxidation that operate by CH activation.


Work-related Helicobacter pylori infection among sewage workers in municipal wastewater treatment plants in Belgium.
Most studies among sewage workers of wastewater treatment plants (WWTPs) have found a higher prevalence of gastrointestinal symptoms than among non-sewage exposed workers. Waterborne transmission of Helicobacter pylori (H pylori) has been hypothesised, as the bacteria can survive into an aqueous environment and has been detected in sewage. A health and hygiene questionnaire has demonstrated a higher prevalence of peptic ulcers among Belgian WWTP operators and maintenance workers than among non-sewage exposed colleagues. To assess the seroprevalence of H pylori infection in Belgian sewage workers at municipal WWTPs, and to determine whether sewage exposure is an important risk factor for acquisition of H pylori and the possible association with gastrointestinal symptoms. A seroprevalence study of H pylori antibodies was conducted among 317 WWTP employees (operators, maintenance workers, laboratory personnel, other job). Information about demographic variables, possible H pylori risk factors, working history, and history of current gastrointestinal symptoms during last 3 months was obtained by a questionnaire. The presence of H pylori IgG was investigated with an ELISA. The results were compared with those of 250 employees of a pharmaceutical company (operators, maintenance workers, laboratory personnel). The prevalence of H pylori IgG antibodies among sewage workers was 16.7% (95% CI 12.6 to 20.8%) compared to 13.6% (95% CI 9.4 to 17.8%) among the control group. In a logistic regression model with controlling for age and educational levels, OR study/control group was 1.02 (95% CI 0.58 to 1.80 with p=0.93). No significant associations were found between the H pylori status and gastrointestinal symptoms, occupational exposures in different tasks, nor with hygienic practices. Our results do not suggest that H pylori infection is a probable cause of part of gastrointestinal symptoms among Flemish municipal sewage workers of WWTPs.


Electron beam computed tomography for the quantification of aortic valvular calcification.
Calcific aortic stenosis is common in the elderly; indeed, 30-60% of patients with mild 'senile' aortic stenosis will progress to severe obstruction. Nonetheless, predictors of progression are incompletely defined, and non-invasive technologies capable of quantifying aortic valve calcium are needed. The reliability of electron beam computed tomography (EBCT) was evaluated for quantification of aortic valve calcium content. Nineteen patients with and without restrictive valve calcification underwent EBCT scanning. Separate calcium scores, 30 s apart, were obtained in all patients, and the Spearman correlation coefficient was calculated between measurements. The relationship between dichotomized mean calcium score and aortic valve area was also investigated. There was excellent correlation between calcium scores (R = 0.99, p = 0.0001), as well as a significant inverse relationship between calcium scores in the upper and lower ranges and aortic valve area (p = 0.002). EBCT can be used for reproducible quantitation of aortic valve calcification. While at their extremes, calcium scores are inversely related to aortic valve area, further evaluation is needed to define the precise nature of this relationship throughout the spectrum of stenosis severity. EBCT holds promise in the longitudinal assessment of valvular calcification progression and its response to potential medical therapies.


On the effects of signal acuity in a multi-alternative model of decision making.
We consider the effects of signal sharpness or acuity on the performance of neural models of decision making. In these models, a vector of signals is presented, and the subject must decide which of the elements of the vector is the largest. McMillen and Holmes ( 2006 ) derived asymptotically optimal tests under the assumption that the elements of the signal vector were all equal except one. In this letter, we consider the case of signals spread around a peak. The acuity is a measure of how strongly peaked the signal is. We find that the optimal test is one in which the detectors are passed through an output layer that encodes knowledge of the possible shapes of the incoming signals. The incorporation of such an output layer can lead to significant improvements in decision-making tasks.


Neural Variability Quenching Predicts Individual Perceptual Abilities.
Neural activity during repeated presentations of a sensory stimulus exhibits considerable trial-by-trial variability. Previous studies have reported that trial-by-trial neural variability is reduced (quenched) by the presentation of a stimulus. However, the functional significance and behavioral relevance of variability quenching and the potential physiological mechanisms that may drive it have been studied only rarely. Here, we recorded neural activity with EEG as subjects performed a two-interval forced-choice contrast discrimination task. Trial-by-trial neural variability was quenched by ∼40% after the presentation of the stimulus relative to the variability apparent before stimulus presentation, yet there were large differences in the magnitude of variability quenching across subjects. Individual magnitudes of quenching predicted individual discrimination capabilities such that subjects who exhibited larger quenching had smaller contrast discrimination thresholds and steeper psychometric function slopes. Furthermore, the magnitude of variability quenching was strongly correlated with a reduction in broadband EEG power after stimulus presentation. Our results suggest that neural variability quenching is achieved by reducing the amplitude of broadband neural oscillations after sensory input, which yields relatively more reproducible cortical activity across trials and enables superior perceptual abilities in individuals who quench more. Variability quenching is a phenomenon in which neural variability across trials is reduced by the presentation of a stimulus. Although this phenomenon has been reported across a variety of animal and human studies, its functional significance and behavioral relevance have been examined only rarely. Here, we report novel empirical evidence from humans revealing that variability quenching differs dramatically across individual subjects and explains to a certain degree why some individuals exhibit better perceptual abilities than others. In addition, we found a strong relationship between variability quenching and suppression of broadband neural oscillations. Together, our results reveal the importance of reproducible cortical activity for enabling better perceptual abilities and suggest a potential underlying mechanism that may explain why variability quenching occurs.


Episodic release of LH in gonadectomized male Japanese quail.
Blood samples were taken every 15 min (for 4.75 h) from six castrated quail on three separate occasions. The birds were first bled while under short days (comprising 8 h light : 16 h darkness per 24 h; 8L : 16D) and, subsequently, after 1 and 20 days under long days of 16L : 8D. The photoperiodic alteration produced a marked increase in the mean plasma LH concentration of each bird and in four instances the rise was evident after only 1 long day. Pulsatile release patterns were not detected in the plasma LH profiles obtained during exposure to short days or after 1 long day but were pronounced in all of the birds after 20 long days. The peaks (n = 19) occurred on average every 83 +/- 14 (S.E.M.) min and had an amplitude of 12.3 +/- 1.2 micrograms/l. After 35 days under 16L : 8D three of the quail were bled more frequently (every 8 min for 2.5 h), allowing the LH pulses to be measured more precisely. Each pulse was composed of a sudden increase in secretion followed by a slower decrease which lasted for approximately 30 min. The development and functional significance of episodic LH release in the quail is discussed.


Multi-institutional study of GRE scores as predictors of STEM PhD degree completion: GRE gets a low mark.
The process of selecting students likely to complete science, technology, engineering and mathematics (STEM) doctoral programs has not changed greatly over the last few decades and still relies heavily on Graduate Record Examination (GRE) scores in most U.S. universities. It has been long debated whether the GRE is an appropriate selection tool and whether overreliance on GRE scores may compromise admission of students historically underrepresented in STEM. Despite many concerns about the test, there are few studies examining the efficacy of the GRE in predicting PhD completion and even fewer examining this question in STEM fields. For the present study, we took advantage of a long-lived collaboration among institutions in the Northeast Alliance for Graduate Education and the Professoriate (NEAGEP) to gather comparable data on GRE scores and PhD completion for 1805 U.S./Permanent Resident STEM doctoral students in four state flagship institutions. We found that GRE Verbal (GRE V) and GRE Quantitative (GRE Q) scores were similar for women who completed STEM PhD degrees and those who left programs. Remarkably, GRE scores were significantly higher for men who left than counterparts who completed STEM PhD degrees. In fact, men in the lower quartiles of GRE V or Q scores finished degrees more often than those in the highest quartile. This pattern held for each of the four institutions in the study and for the cohort of male engineering students across institutions. GRE scores also failed to predict time to degree or to identify students who would leave during the first year of their programs. Our results suggests that GRE scores are not an effective tool for identifying students who will be successful in completing STEM doctoral programs. Considering the high cost of attrition from PhD programs and its impact on future leadership for the U.S. STEM workforce, we suggest that it is time to develop more effective and inclusive admissions strategies.


Central role of the adipocyte in the metabolic syndrome.
Insulin resistance is associated with a plethora of chronic illnesses, including Type 2 diabetes, dyslipidemia, clotting dysfunction, and colon cancer. The relationship between obesity and insulin resistance is well established, and an increase in obesity in Western countries is implicated in increased incidence of diabetes and other diseases. Central, or visceral, adiposity has been particularly associated with insulin resistance; however, the mechanisms responsible for this association are unclear. Our laboratory has been studying the physiological mechanisms relating visceral adiposity and insulin resistance. Moderate fat feeding of the dog yields a model reminiscent of the metabolic syndrome, including visceral adiposity, hyperinsulinemia, and insulin resistance. We propose that insulin resistance of the liver derives from a relative increase in the delivery of free fatty acids (FFA) from the omental fat depot to the liver (via the portal vein). Increased delivery results from 1) more stored lipids in omental depot, 2) severe insulin resistance of the central fat depot, and 3) possible regulation of visceral lipolysis by the central nervous system. The significance of portal FFA delivery results from the importance of FFA in the control of liver glucose production. Insulin regulates liver glucose output primarily via control of adipocyte lipolysis. Thus, because FFA regulate the liver, it is expected that visceral adiposity will enhance delivery of FFA to the liver and make the liver relatively insulin resistant. It is of interest how the intact organism compensates for insulin resistance secondary to visceral fat deposition. While part of the compensation is enhanced B-cell sensitivity to glucose, an equally important component is reduced liver insulin clearance, which allows for a greater fraction of B-cell insulin secretion to bypass liver degradation, to enter the systemic circulation, and to result in hyperinsulinemic compensation. The signal(s) resulting in B-cell up-regulation and reduced liver insulin clearance with visceral adiposity is (are) unknown, but it appears that the glucagon-like peptide (GLP-1) hormone plays an important role. The integrated response of the organism to central adiposity is complex, involving several organs and tissue beds. An investigation into the integrated response may help to explain the features of the metabolic syndrome.


Homeostasis model assessment of insulin resistance does not seem to predict response to telaprevir in chronic hepatitis C in the REALIZE trial.
Baseline homeostasis model assessment-estimated insulin resistance (HOMA-IR), a marker for insulin resistance, has been associated with poor virologic response to peginterferon alpha/ribavirin (PR) in chronic hepatitis C. We evaluated the association between baseline HOMA-IR and pretreatment factors on sustained virologic response (SVR) to telaprevir (TVR) in genotype 1 patients with hepatitis C and prior peginterferon/ribavirin (PR) treatment failure. Patients were randomized to 12 weeks of TVR (750 mg q8h) plus peginterferon (180 μg/week) and ribavirin (1,000-1,200 mg/day) (with or without a 4-week lead-in) followed by PR, or PR alone (PR48), for 48 weeks. Univariate and multiple logistic regression analyses explored the prognostic significance of baseline HOMA-IR alone and adjusted for other pretreatment factors and SVR. The TVR arms were pooled for the purposes of this analysis. In all, 662 patients were randomized; 578 had baseline HOMA-IR and other prognostic data and were included in this analysis. Median baseline HOMA-IR was 2.6 (interquartile range [IQR] 1.7-4.3); 207 (36%), 206 (36%), and 165 (29%) patients had baseline HOMA-IR <2, 2 to <4, and ≥ 4, respectively. Male gender, higher body mass index, triglycerides, gamma-glutamyl transpeptidase, maximum alanine aminotransferase/aspartate aminotransferase, and fibrosis stage were associated with higher baseline HOMA-IR. Baseline HOMA-IR was associated with SVR in univariate analysis, but not after adjustment for other baseline prognostic factors (TVR: OR = 0.95, 95% confidence interval [CI]: 0.71,1.29; PR48: 0.60; 95% CI: 0.25,1.43). In patients with prior PR treatment failure, baseline HOMA-IR correlated with SVR in univariate but not multivariate analyses, suggesting other factors have a more direct causal relationship with virologic response to TVR-based therapy than HOMA-IR.


Next generation multilocus sequence typing (NGMLST) and the analytical software program MLSTEZ enable efficient, cost-effective, high-throughput, multilocus sequencing typing.
Multilocus sequence typing (MLST) has become the preferred method for genotyping many biological species, and it is especially useful for analyzing haploid eukaryotes. MLST is rigorous, reproducible, and informative, and MLST genotyping has been shown to identify major phylogenetic clades, molecular groups, or subpopulations of a species, as well as individual strains or clones. MLST molecular types often correlate with important phenotypes. Conventional MLST involves the extraction of genomic DNA and the amplification by PCR of several conserved, unlinked gene sequences from a sample of isolates of the taxon under investigation. In some cases, as few as three loci are sufficient to yield definitive results. The amplicons are sequenced, aligned, and compared by phylogenetic methods to distinguish statistically significant differences among individuals and clades. Although MLST is simpler, faster, and less expensive than whole genome sequencing, it is more costly and time-consuming than less reliable genotyping methods (e.g. amplified fragment length polymorphisms). Here, we describe a new MLST method that uses next-generation sequencing, a multiplexing protocol, and appropriate analytical software to provide accurate, rapid, and economical MLST genotyping of 96 or more isolates in single assay. We demonstrate this methodology by genotyping isolates of the well-characterized, human pathogenic yeast Cryptococcus neoformans.


Pairing of homologous regions in the mouse genome is associated with transcription but not imprinting status.
Although somatic homologous pairing is common in Drosophila it is not generally observed in mammalian cells. However, a number of regions have recently been shown to come into close proximity with their homologous allele, and it has been proposed that pairing might be involved in the establishment or maintenance of monoallelic expression. Here, we investigate the pairing properties of various imprinted and non-imprinted regions in mouse tissues and ES cells. We find by allele-specific 4C-Seq and DNA FISH that the Kcnq1 imprinted region displays frequent pairing but that this is not dependent on monoallelic expression. We demonstrate that pairing involves larger chromosomal regions and that the two chromosome territories come close together. Frequent pairing is not associated with imprinted status or DNA repair, but is influenced by chromosomal location and transcription. We propose that homologous pairing is not exclusive to specialised regions or specific functional events, and speculate that it provides the cell with the opportunity of trans-allelic effects on gene regulation.


Pre-B cell colony-enhancing factor inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis.
Pre-B cell colony-enhancing factor (PBEF) is a highly conserved 52-kDa protein, originally identified as a growth factor for early stage B cells. We show here that PBEF is also upregulated in neutrophils by IL-1beta and functions as a novel inhibitor of apoptosis in response to a variety of inflammatory stimuli. Induction of PBEF occurs 5-10 hours after LPS exposure. Prevention of PBEF translation with an antisense oligonucleotide completely abrogates the inhibitory effects of LPS, IL-1, GM-CSF, IL-8, and TNF-alpha on neutrophil apoptosis. Immunoreactive PBEF is detectable in culture supernatants from LPS-stimulated neutrophils, and a recombinant PBEF fusion protein inhibits neutrophil apoptosis. PBEF is also expressed in neutrophils from critically ill patients with sepsis in whom rates of apoptosis are profoundly delayed. Expression occurs at higher levels than those seen in experimental inflammation, and a PBEF antisense oligonucleotide significantly restores the normal kinetics of apoptosis in septic polymorphonuclear neutrophils. Inhibition of apoptosis by PBEF is associated with reduced activity of caspases-8 and -3, but not caspase-9. These data identify PBEF as a novel inflammatory cytokine that plays a requisite role in the delayed neutrophil apoptosis of clinical and experimental sepsis.


Adult norms for a commercially available Nine Hole Peg Test for finger dexterity.
The Nine Hole Peg Test is commonly used by occupational therapists as a simple, quick assessment for finger dexterity. The purpose of this study was to evaluate the interrater and test-retest reliability of the commercially available Smith & Nephew Rehabilitation Division version of the Nine Hole Peg Test, and to establish new adult norms for the Nine Hole Peg Test for finger dexterity utilizing this particular version. Two of the researchers established interrater and test-retest reliability by evaluating 25 occupational therapy student volunteers. Seven hundred and three subjects, ranging in age from 21 to 71+ years, were tested to establish norms, using the standard protocol. Results showed high interrater reliability and only moderate test-retest reliability. Scores obtained by using the commercially available version were not statistically different from previously published norms (Mathiowetz, Weber, Kashman, & Volland, 1985). This study supports the original norms and further assists occupational therapists to evaluate dexterity accurately.


The degree of antenatal ventriculomegaly is related to pediatric neurological morbidity.
Our hypothesis was that the degree of antenatally diagnosed cerebral ventriculomegaly is related to aneuploidy, perinatal mortality and long-term neurological morbidity. Ninety-one cases of ventriculomegaly identified from 1 June 1994 to 1 July 1999 were examined for prenatal, intrapartum and neonatal complications. Pediatric follow-up was reviewed for infants with ventriculomegaly from birth up to as long as 4 years. Minor neurological morbidity was defined as a score of 70-80 on the clinical adaptive test/clinical linguistic and auditory milestone scale and included mild motor or language delay. Major morbidity included a score of < 70, evidence of cerebral palsy, or seizure disorder. The incidence of neurological complications was compared, on the basis of the degree of ventriculomegaly, with group 1 being > 10-15 mm and group 2 being > 15 mm. Twenty-seven cases (18 with neural tube defects and nine with holoprosencephaly) were excluded. Among the remaining 64 patients, 39 had a ventricular diameter of > 10-15 mm and comprised group 1. Five of the 39 cases (12.8%), all with other ultrasound anomalies, elected to terminate. The incidence of aneuploidy in group 1 was 14.2%. Among the 19 cases with isolated ventriculomegaly, 17 (89%) were normal and two (11%) had minor neurological morbidity. In group 1 there were two cases associated with cytomegalovirus (CMV) infection. Of the 25 cases in group 2, eight (32%), all with other ultrasound anomalies, elected to terminate. The incidence of aneuploidy in group 2 was 17.4%. For the nine cases with isolated ventriculomegaly of > 15 mm, one (11%) was normal (p < 0.001), five (56%) had minor neurological morbidity requiring a ventriculoperitoneal shunt (p = 0.035), and three (33%) had major neurological morbidity (p = 0.045) when compared to cases of isolated ventriculomegaly in group 1. There was one case of CMV infection in group 2. All perinatal deaths in both groups were associated with other anomalies. Amniocentesis to determine karyotype and the presence of CMV is warranted for all cases of ventriculomegaly of > 10 mm. The degree of antenatal ventriculomegaly is related to pediatric neurological morbidity and, when it is > 15 mm, it is associated with an increase in abnormal neurological development.


The epidemiology of fungal infections in patients with hematologic malignancies: the SEIFEM-2004 study.
The aim of this study was to evaluate the incidence and outcome of invasive fungal infections (IFI) in patients with hematologic malignancies. This was a retrospective cohort study of patients admitted between 1999 and 2003 to 18 hematology wards in Italy. Each participating center provided information on all patients with newly diagnosed hematologic malignancies admitted during the survery period and on all episodes of IFI experienced by these patients. The cohort was formed of 11,802 patients with hematologic malignacies: acute leukemia (myeloid 3012, lymphoid 1173), chronic leukemia (myeloid 596, lymphoid 1104), lymphoma (Hodgkin's 844, non-Hodgkin's 3457), or multiple myeloma (1616). There were 538 proven or probable IFI (4.6%); 373 (69%) occurred in patients with acute myeloid leukemia. Over half (346/538) were caused by molds (2.9%), in most cases Aspergillus spp. (310/346). The 192 yeast infections (1.6%) included 175 cases of candidemia. Overall and IFI-attributable mortality rates were 2% (209/11802) and 39% (209/538), respectively. The highest IFI-attributable mortality rates were associated with zygomycosis (64%) followed by fusariosis (53%), aspergillosis (42%), and candidemia (33%). Patients with hematologic malignancies are currently at higher risk of IFI caused by molds than by yeasts, and the incidence of IFI is highest among patients with acute myeloid leukemia. Aspergillus spp are still the most common pathogens, followed by Candida spp. Other agents are rare. The attributable mortality rate for aspergillosis has dropped from 60-70% to approximately 40%. Candidemia-related mortality remains within the 30-40% range reported in literature although the incidence has decreased.


Cyclic 3',5'-AMP relay in Dictyostelium discoideum IV. Recovery of the cAMP signaling response after adaptation to cAMP.
In dictyoselium discoideum, an increase in extracellular cAMP activates adenylate cyclase, leading to an increase in intracellular cAMP and the rate of cAMP secretion. Cells adapt to any constant cAMP stimulus after several minutes, but still respond to an increase in the concentration of the stimulus. We have now characterized the decay of adaptation (deadaptation) after the removal of cAMP stimuli. Levels of adaptation were established by the perfusion of [(3)H]adenosine-labeled amoebae with a defined cAMP stimulus. After a variable recovery period, the magnitude of the signaling response to a second stimulus was measured; its attenuation was taken as a measure of residual adaption to the first stimulus. The level of adaptation established by the first stimulus depended on both its magnitude and duration. Deadaptation began as soon as the first stimulus was removed. The magnitude of the response to the second stimulus increased with the recovery time in a first-order fashion, with a t(1/2)=3-4 min for stimuli of 10(-8) M to 10(-5) M cAMP. Responses to test stimuli, although reduced in magnitude, had an accelerated time-course when they closely followed a prior response that had not completely subsided. This effect is called priming; we believe it reveals a reversible, rate-limiting step that modulates the onset and termination of the signaling responses of amoebae that have not recently responded to a cAMP stimulus. We have suggested that the cAMP signaling response is controlled by two antagonistic cellular processes, excitation and adaptation. The data reported here imply that both the rate of rise in the adaptation process and the final level reached depend on the occupancy of cAMP surface receptors and that the decay of adaptation when external cAMP is removed proceeds with first-order kinetics.


A literature review on flow-rate variability in neonatal IV therapy.
To provide an overview of factors influencing the flow rate in intravenous (IV) therapy for newborns. We conducted a review of the literature from 1980 to 2011 in PubMed and Web of Knowledge. Articles focusing on flow-rate variability and possible complications due to flow-rate variability were included. Forty-one articles were selected for this review. IV therapy in (preterm) neonates is prone to significant start-up delays and flow-rate variability. The sudden changes in the volume delivered to (preterm) neonates may have serious consequences. Low preprogrammed flow rates, total compliance, and volume of the IV administration set, the presence or absence of antisiphon valves or inline filters and the vertical displacement of syringe pumps all contribute to flow-rate variability in IV therapy for neonates. Flow-rate variability in IV therapy and its clinical relevance are due to the preprogrammed flow rate, the hydrostatic pressure changes, the complete IV administration set compliance and the type of substances supplied to the patient. To improve IV therapy, the internal compliance of the complete IV administration set should be minimized and the highest possible preprogrammed flow rate should be used in combination with small syringes and low-resistance valves.


3D plasmonic nanoantennas integrated with MEA biosensors.
Neuronal signaling in brain circuits occurs at multiple scales ranging from molecules and cells to large neuronal assemblies. However, current sensing neurotechnologies are not designed for parallel access of signals at multiple scales. With the aim of combining nanoscale molecular sensing with electrical neural activity recordings within large neuronal assemblies, in this work three-dimensional (3D) plasmonic nanoantennas are integrated with multielectrode arrays (MEA). Nanoantennas are fabricated by fast ion beam milling on optical resist; gold is deposited on the nanoantennas in order to connect them electrically to the MEA microelectrodes and to obtain plasmonic behavior. The optical properties of these 3D nanostructures are studied through finite elements method (FEM) simulations that show a high electromagnetic field enhancement. This plasmonic enhancement is confirmed by surface enhancement Raman spectroscopy of a dye performed in liquid, which presents an enhancement of almost 100 times the incident field amplitude at resonant excitation. Finally, the reported MEA devices are tested on cultured rat hippocampal neurons. Neurons develop by extending branches on the nanostructured electrodes and extracellular action potentials are recorded over multiple days in vitro. Raman spectra of living neurons cultured on the nanoantennas are also acquired. These results highlight that these nanostructures could be potential candidates for combining electrophysiological measures of large networks with simultaneous spectroscopic investigations at the molecular level.


Systematic colocalization errors between acridine orange and EGFP in astrocyte vesicular organelles.
Dual-color imaging of acridine orange (AO) and EGFP fused to a vesicular glutamate transporter or the vesicle-associated membrane proteins 2 or 3 has been used to visualize a supposedly well-defined subpopulation of glutamatergic astrocytic secretory vesicles undergoing regulated exocytosis. However, AO metachromasy results in the concomitant emission of green and red fluorescence from AO-stained tissue. Therefore, the question arises whether AO and EGFP fluorescence can be distinguished reliably. We used evanescent-field imaging with spectral fluorescence detection as well as fluorescence lifetime imaging microscopy to demonstrate that green fluorescent AO monomers inevitably coexist with red fluorescing AO dimers, at the level of single astroglial vesicles. The green monomer emission spectrally overlaps with that of EGFP and produces a false apparent colocalization on dual-color images. On fluorophore abundance maps calculated from spectrally resolved and unmixed single-vesicle spectral image stacks, EGFP is obscured by the strong green monomer fluorescence, precluding the detection of EGFP. Hence, extreme caution is required when deriving quantitative colocalization information from images of dim fluorescing EGFP-tagged organelles colabeled with bright and broadly emitting dyes like AO. We finally introduce FM4-64/EGFP dual-color imaging as a remedy for imaging a distinct population of astroglial fusion-competent secretory vesicles.


Voluntary counseling and testing (VCT) services and their contribution to access to HIV diagnosis in Brazil.
The Voluntary Counseling and Testing (VCT) Network was implemented in Brazil in the 1980s to promote anonymous and confidential access to HIV diagnosis. As a function of the population and dimensions of the local epidemic, the study assessed the network's coverage, using data from a self-applied questionnaire and data from the Information Technology Department of the Unified National Health System (SUS), UNDP, and National STD/AIDS Program. The Student t test was used for comparison of means and the chi-square test for proportions. Brazil has 383 VCT centers, covering 48.9% of the population and 69.2% of the AIDS cases. The network has been implemented predominantly in regions where the epidemic shows a relevant presence, but 85.3% of the cities with high HIV incidence lack VCT centers; absence of VCT was associated with more limited health infrastructure and worse social indicators. A slowdown in expansion of the network was observed, with VCT Centers implemented on average 16 years after the first AIDS case in the given municipality. The number of HIV tests performed under the SUS is 2.3 times higher in cities with VCT centers. The network's scope is limited, thus minimizing the contribution by these services to the supply of HIV diagnosis in Brazil.


Modulation of aflatoxin metabolism, aflatoxin-N7-guanine formation, and hepatic tumorigenesis in rats fed ethoxyquin: role of induction of glutathione S-transferases.
The effects of dietary administration of ethoxyquin (EQ) on aflatoxin B1 (AFB1) metabolism, DNA adduct formation and removal, and hepatic tumorigenesis were examined in male Fischer rats. Rats were fed a semipurified diet containing 0.4% EQ for 1 wk, gavaged with 250 micrograms of AFB1 per kg 5 times a wk during the next 2 wk, and, finally, restored to the control diet 1 wk after cessation of dosing. At 4 mo, focal areas of hepatocellular alteration were identified and quantitated by staining sections of liver for gamma-glutamyl transpeptidase. Treatment with EQ reduced by greater than 95% both area and volume of liver occupied by gamma-glutamyl transpeptidase-positive foci. Utilizing the same multiple dosing protocol, patterns of covalent modifications of DNA by AFB1 were determined. EQ produced a dramatic reduction in the binding of AFB1 to hepatic DNA: 18-fold initially and 3-fold at the end of the dosing period. Although binding was detectable at 3 and 4 mo postdosing, no effect of EQ was observed, suggesting that these persistent adducts are not of primary relevance to AFB1 carcinogenesis. Analysis of nucleic acid bases by high-performance liquid chromatography revealed no qualitative differences in adduct species between treatment groups. The inhibitory effect of EQ on AFB1 binding to DNA and tumorigenesis appears related to induction of detoxication enzymes. Rats fed 0.4% EQ for 7 days showed a 5-fold increase in hepatic cytosolic glutathione S-transferase (GST)-specific activities. Multiple molecular forms of GST were induced, and concomitant elevations in messenger RNA levels coding for the synthesis of GST subunits were observed. Correspondingly, biliary elimination of AFB1-glutathione conjugate was increased 4.5-fold in animals on the EQ diet during the first 2 h following p.o. administration of 250 micrograms of AFB1 per kg. Thus, induction by EQ of enzymes important to AFB1 detoxication, such as GST, can lead to enhanced carcinogen elimination, as well as reductions of AFB1-DNA adduct formation and subsequent expression of preneoplastic lesions, and, ultimately, neoplasia.


The trappin gene family: proteins defined by an N-terminal transglutaminase substrate domain and a C-terminal four-disulphide core.
Recently, several new genes have been discovered in various species which are homologous to the well-characterized human epithelial proteinase inhibitor elafin/SKALP (skin-derived anti-leukoproteinase). Because of the high degree of conservation and the similarities in genomic organization, we propose that these genes belong to a novel gene family. At the protein level, the family members are defined by: (1) an N-terminal domain consisting of a variable number of repeats with the consensus sequence Gly-Gln-Asp-Pro-Val-Lys that can act as an anchoring motif by transglutaminase cross-linking, and (2) a C-terminal four-disulphide core or whey acidic protein (WAP) domain, which harbours a functional motif involved in binding of proteinases and possibly other proteins. We have proposed the name trappin gene family as a unifying nomenclature for this group of proteins (trappin is an acronym for TRansglutaminase substrate and wAP domain containing ProteIN, and refers to its functional property of 'getting trapped' in tissues by covalent cross-linking). Analysis of the trappin family members shows extensive diversification in bovidae and suidae, whereas the number of primate trappins is probably limited. Recent biochemical and cell biological data on the human trappin family member elafin/SKALP suggest that this molecule is induced in epidermis by cellular stress. We hypothesize that trappins play an important role in the regulation of inflammation and in protection against tissue damage in stratified epithelia.


Associations of cannabis use disorder with cognition, brain structure, and brain function in African Americans.
Although previous studies have highlighted associations of cannabis use with cognition and brain morphometry, critical questions remain with regard to the association between cannabis use and brain structural and functional connectivity. In a cross-sectional community sample of 205 African Americans (age 18-70) we tested for associations of cannabis use disorder (CUD, n = 57) with multi-domain cognitive measures and structural, diffusion, and resting state brain-imaging phenotypes. Post hoc model evidence was computed with Bayes factors (BF) and posterior probabilities of association (PPA) to account for multiple testing. General cognitive functioning, verbal intelligence, verbal memory, working memory, and motor speed were lower in the CUD group compared with non-users (p < .011; 1.9 < BF < 3,217). CUD was associated with altered functional connectivity in a network comprising the motor-hand region in the superior parietal gyri and the anterior insula (p < .04). These differences were not explained by alcohol, other drug use, or education. No associations with CUD were observed in cortical thickness, cortical surface area, subcortical or cerebellar volumes (0.12 < BF < 1.5), or graph-theoretical metrics of resting state connectivity (PPA < 0.01). In a large sample collected irrespective of cannabis used to minimize recruitment bias, we confirm the literature on poorer cognitive functioning in CUD, and an absence of volumetric brain differences between CUD and non-CUD. We did not find evidence for or against a disruption of structural connectivity, whereas we did find localized resting state functional dysconnectivity in CUD. There was sufficient proof, however, that organization of functional connectivity as determined via graph metrics does not differ between CUD and non-user group.


What is "functional impairment"? Disentangling disability from clinical significance.
The formulation of disability (or "functional impairment") is currently not operationalized in either the ICD or the DSM. In the DSM system, making a diagnosis depends on a conjoint assessment of symptoms and functioning, whereas the ICD keeps the disability construct separate from the diagnosis of mental disorders. We need an internationally agreed conceptualization between ICD and DSM in terms of better operationalization of disease and disability components. No functioning or disability should appear as part of the threshold of the diagnosis in either system.


Challenges in the treatment of elderly patients with primary central nervous system lymphoma.
Approximately 50% of all patients with primary central nervous system lymphoma (PCNSL) are 60 years or older and may therefore be considered as elderly. Although the diagnostic work-up is basically the same in young and in elderly patients, therapeutic strategies vary considerably. Here, we review the characteristics of elderly PCNSL patients with a particular focus on advances in the optimization of treatment regimens. Age has been repeatedly confirmed as a major therapy-independent negative prognostic factor. Benefit from treatment and the tolerability of tumor-specific therapy, particularly whole-brain radiotherapy, are significantly lower in the elderly patients. Still, for patients with newly diagnosed PCNSL, several studies emphasized the indisputable role of high-dose methotrexate as backbone for any therapy regimen also in elderly patients. However, the durability of responses to primary chemotherapy is significantly shorter than in young patients. Recent data from a randomized phase II study for elderly PCNSL patients suggest that the combination of high-dose methotrexate, procarbazine, vincristine and cytarabine is superior to methotrexate in combination with temozolomide. Current efforts aim at treating elderly PCNSL patients within clinical trials that are specifically designed for this group of patients. Determining adapted consolidation and/or maintenance treatment to improve disease control in responding patients are the main challenges to be faced by future trials. Together with a better understanding of age-specific changes in the biology of PCNSL, this will pave the road for elderly tailored therapies.


Structure and Composition Define Immunorecognition of Nucleic Acid Nanoparticles.
Nucleic acid nanoparticles (NANPs) have evolved as a new class of therapeutics with the potential to detect and treat diseases. Despite tremendous advancements in NANP development, their immunotoxicity, one of the major impediments in clinical translation of traditional therapeutic nucleic acids (TNAs), has never been fully characterized. Here, we describe the first systematically studied immunological recognition of 25 representative RNA and DNA NANPs selected to have different design principles and physicochemical properties. We discover that, unlike traditional TNAs, NANPs used without a delivery carrier are immunoquiescent. We show that interferons (IFNs) are the key cytokines triggered by NANPs after their internalization by phagocytic cells, which agrees with predictions based on the experiences with TNAs. However, in addition to type I IFNs, type III IFNs also serve as reliable biomarkers of NANPs, which is usually not characteristic of TNAs. We show that overall immunostimulation relies on NANP shapes, connectivities, and compositions. We demonstrate that, like with traditional TNAs, plasmacytoid dendritic cells serve as the primary interferon producers among all peripheral blood mononuclear cells treated with NANPs, and scavenger receptor-mediated uptake and endosomal Toll-like receptor signaling are essential for NANP immunorecognition. The TLR involvement, however, is different from that expected for traditional TNA recognition. Based on these results, we suggest that NANP technology may serve as a prototype of auxiliary molecular language for communication with the immune system and the modulation of immune responses.


#PlasticSurgery.
Social media use is growing inexorably, and there is public appetite for evidence-based information. Little is known about engagement by plastic surgeons with social media. The aim of this study was to examine posting about plastic surgery on Twitter, to best inform how board-certified plastic surgeons could use the hashtag #PlasticSurgery as a tool to educate patients and the public. A prospective analysis of 2880 "tweets" containing the words "plastic surgery" was performed. The following were assessed: identity of author, use of the hashtag #PlasticSurgery, subject matter, whether link to study was provided, and whether posts by surgeons were self-promotional or educational. Social media posting about plastic surgery is dominated by the public, accounting for 70.6 percent of posts versus only 6.0 percent by plastic surgeons. Only 5.4 percent of all tweets contained the hashtag #PlasticSurgery, although almost half of those that did were by plastic surgeons. Of these, 61.3 percent of posts by plastic surgeons were about aesthetic surgery; additional posts were about basic science, patient safety, and reconstruction (13.9, 4.0, and 2.3 percent, respectively). Eighteen scientific articles were referenced, with a link to the Journal site posted in two tweets. Of posts by plastic surgeons, 37.0 percent were self-promotional. The American Society of Plastic Surgeons and its Journal have recognized that social media may be used to educate and engage. Board-certified plastic surgeons have a great opportunity to promote evidence-based plastic practice by means of #PlasticSurgery in the interests of supporting patients and the profession.


Reasons for outpatient referrals from generalists to specialists.
To determine the relative importance of medical and nonmedical factors influencing generalists' decisions to refer, and of the factors that might avert unnecessary referrals. Prospective survey of all referrals from generalists to subspecialists over a 5-month period. University hospital outpatient clinics. Fifty-seven staff physicians in general internal medicine, family medicine, dermatology, orthopedics, gastroenterology, and rheumatology. For each referral, the generalist rated a number of medical and nonmedical reasons for referral, as well as factors that may have helped avert the referral; the specialist seeing the patient then rated the appropriateness, timeliness, and complexity of the referral. Both physicians rated the potential avoidability of the referral by telephone consultation. Generalists were influenced by a combination of both medical and nonmedical reasons for 76% of the referrals, by only medical reasons in 20%, and by only nonmedical reasons in 3%. In 33% of all referrals, generalists felt that training in simple procedures or communication with a generalist or specialist colleague would have allowed them to avoid referral. Specialists felt that the vast majority of referrals were timely (as opposed to premature or delayed) and of average complexity. Although specialists rated most referrals as appropriate, 30% were rated as possibly appropriate or inappropriate. Generalists and specialists failed to agree on the avoidability of 34% of referrals. Generalists made most referrals for a combination of medical and nonmedical reasons, and many referrals were considered avoidable. Increasing procedural training for generalists and enhancing informal channels of communication between generalists and subspecialists might result in more appropriate referrals at lower cost.


Genome-wide association mapping for flowering and maturity in tropical soybean: implications for breeding strategies.
Knowledge of the genetic architecture of flowering and maturity is needed to develop effective breeding strategies in tropical soybean. The aim of this study was to identify haplotypes across multiple environments that contribute to flowering time and maturity, with the purpose of selecting desired alleles, but maintaining a minimal impact on yield-related traits. For this purpose, a genome-wide association study (GWAS) was undertaken to identify genomic regions that control days to flowering (DTF) and maturity (DTM) using a soybean association mapping panel genotyped for single nucleotide polymorphism (SNP) markers. Complementarily, yield-related traits were also assessed to discuss the implications for breeding strategies. To detect either stable or specific associations, the soybean cultivars (N = 141) were field-evaluated across eight tropical environments of Brazil. Seventy-two and forty associations were significant at the genome-wide level relating respectively to DTM and DTF, in two or more environments. Haplotype-based GWAS identified three haplotypes (Gm12_Hap12; Gm19_Hap42 and Gm20_Hap32) significantly co-associated with DTF, DTM and yield-related traits in single and multiple environments. These results indicate that these genomic regions may contain genes that have pleiotropic effects on time to flowering, maturity and yield-related traits, which are tightly linked with multiple other genes with high rates of linkage disequilibrium.


Protein disulfide isomerase knockdown-induced cell death is cell-line-dependent and involves apoptosis in MCF-7 cells.
Protein disulfide isomerase (PDI) is a multifunctional protein that catalyzes disulfide bond formation and assists protein folding, as well as being a structural subunit of microsomal triglyceride transfer protein (MTP) and prolyl 4-hydroxylase (P4HD), and an estrogen and thyroid hormone-binding protein. Previous reports indicate that some endocrine-disrupting chemicals (EDCs) bind to PDI and disturb its functions, and we executed PDI-knockdown to examine the effects of dysfunction of PDI. In this study, the effects of PDI-knockdown were compared among three cell lines: MCF-7, SH-SY5Y and HeLa. PDI-knockdown induced different levels of cytotoxicity among these cell lines. In MCF-7 cells, PDI-knockdown activated apoptotic signaling, causing cytochrome c release from mitochondria and activation of caspase-9, caspase-6, caspase-7 and poly[ADP-ribose]polymerase-1, and the cytotoxicity induced by PDI-knockdown was suppressed by a pan-caspase inhibitor, z-VAD-fmk. These data suggest that cell death induced by PDI-knockdown is caspase-dependent apoptosis in MCF-7 cells.


Restrictive eating across a spectrum from healthy to unhealthy: behavioral and neural mechanisms.
Restriction of food intake is a central feature of anorexia nervosa (AN) and other eating disorders, yet also occurs in the absence of psychopathology. The neural mechanisms of restrictive eating in health and disease are unclear. This study examined behavioral and neural mechanisms associated with restrictive eating among individuals with and without eating disorders. Dietary restriction was examined in four groups of women (n = 110): healthy controls, dieting healthy controls, patients with subthreshold (non-low weight) AN, and patients with AN. A Food Choice Task was administered during fMRI scanning to examine neural activation associated with food choices, and a laboratory meal was conducted. Behavioral findings distinguished between healthy and ill participants. Healthy individuals, both dieting and non-dieting, chose significantly more high-fat foods than patients with AN or subthreshold AN. Among healthy individuals, choice was primarily influenced by tastiness, whereas, among both patient groups, healthiness played a larger role. Dorsal striatal activation associated with choice was most pronounced among individuals with AN and was significantly associated with selecting fewer high-fat choices in the task and lower caloric intake in the meal the following day. A continuous spectrum of behavior was suggested by the increasing amount of weight loss across groups. Yet, data from this Food Choice Task with fMRI suggest there is a behavioral distinction between illness and health, and that the neural mechanisms underlying food choice in AN are distinct. These behavioral and neural mechanisms of restrictive eating may be useful targets for treatment development.


Genetic Landscape of Congenital Myasthenic Syndromes From Turkey: Novel Mutations and Clinical Insights.
Congenital myasthenic syndromes are clinically and genetically heterogeneous disorders of neuromuscular transmission. Most are treatable, but certain subtypes worsen with cholinesterase inhibitors. This underlines the importance of genetic diagnosis. Here, the authors report on cases with genetically proven congenital myasthenic syndromes from Turkey. The authors retrospectively reviewed their experience of all patients with congenital myasthenic syndromes, referred over a 5-year period (2011-2016) to the Child Neurology Department of Dokuz Eylül University, Izmir, Turkey. In addition, PubMed was searched for published cases of genetically proven congenital myasthenic syndromes originating from Turkey. In total, the authors identified 43 (8 new patients, 35 recently published patients) cases. Defects in the acetylcholine receptor (n = 15; 35%) were the most common type, followed by synaptic basal-lamina associated (n = 14; 33%) and presynaptic syndromes (n = 10; 23%). The authors had only 3 cases (7%) who had defects in endplate development. One patient had mutation GFPT1 gene (n = 1; 2%). Knowledge on congenital myasthenic syndromes and related genes in Turkey will lead to prompt diagnosis and treatment of these rare neuromuscular disorders.


The association of statin use and gonado-sexual function in women: a retrospective cohort analysis.
It has been hypothesized that statins reduce sex hormone biosynthesis through hepatic inhibition of cholesterol synthesis, which is a precursor of androstenedione and estradiol. Such a reduction has been associated with menstrual irregularities, menopausal disorders, infertility, and low libido, but studies are conflicting. Few studies have evaluated the clinical effects of statins on gonadal-sexual function in women. To compare the risk of gonado-sexual dysfunction in statin users vs. nonusers. This was a retrospective cohort study of all female, adult patients (30-85 years) enrolled in the Tricare Prime/Plus San Antonio catchment area. Using 79 baseline characteristics, we created a propensity score-matched cohort of statin users and nonusers. The study duration was divided into a baseline period (October 1, 2003 to September 30, 2005) to describe patient baseline characteristics and a follow-up period (October 1, 2005 to March 1, 2012) to determine patient outcomes. Statin users were defined as those prescribed a statin for ≥3 months between October 1, 2004 and September 30, 2005. Logistic regression was used to determine the association of statin use with patient outcomes. Outcomes included menstrual disorders, menopausal disorders, infertility, and ovarian/sexual dysfunction during the follow-up period. Outcomes were identified using inpatient or outpatient International Classification of Diseases, Ninth Revision, Clinical Modification codes as defined by the Agency for Healthcare Research and Quality's Clinical Classifications Software. Of 22,706 women who met study criteria, we propensity score-matched 2,890 statin users with 2,890 nonusers; mean age 58 ± 12 years. Statin use was not significantly associated with menstrual disorders (OR 0.97; 95% CI 0.81-1.16), menopausal disorders (OR 0.92; 95% CI 0.83-1.02), infertility (OR 0.79; 95% CI 0.36-1.73), or ovarian/sexual dysfunction (OR 1.18; 95% CI 0.83-1.70). Statin use was not associated with higher risk of gonado-sexual dysfunction in women.


A scoping review on the challenges, improvement programs, and relevant output metrics for neurotrauma services in major trauma centers.
For a neurotrauma unit to be defined as a structured neurotrauma service (NS) the following criteria must be satisfied: A dedicated neurointensive care unit, endovascular neuroradiology, in-hospital neurorehabilitation unit and helicopter platform within the context of a Level I trauma center. Designing an effective NS can be challenging, particularly when considering the different priorities and resources of countries across the globe. In addition the impact on clinical outcomes is not clearly established. A scoping review of the literature spanning from 2000 to 2020 meant to identify protocols, guidelines, and best practices for the management of traumatic brain injury (TBI) in NS was conducted on the US National Library of Medicine and National Institute of Health databases. Limited evidence is available regarding quantitative and qualitative metrics to assess the impact of NSs and specialist follow-up clinics on patients' outcome. Of note, the available literature used to lack detailed reports for: (a) Geographical clusters, such as low-to-middle income countries (LMIC); (b) clinical subgroups, such as mild TBI; and (c) long-term management, such as rehabilitation services. Only in the last few years more attention has been paid to those research topics. NSs can positively impact the management of the broad spectrum of TBI in different clinical settings; however more research on patients' outcomes and quality of life metrics is needed to establish their efficacy. The collaboration of global clinicians and the development of international guidelines applicable also to LMIC are warranted.


IL-4 induces eotaxin production in corneal keratocytes but not in epithelial cells.
In severe allergic eye diseases, the breakdown of epithelial barrier function can lead to severe corneal damage such as erosions or ulcers which often resist treatment. Although eosinophils are thought to play a crucial role in corneal tissue damage in severe ocular allergy, the mechanisms of eosinophil recruitment to the cornea has not been fully clarified. Eotaxin has been found in tears of severe allergic patients with corneal ulcer. In this study, we investigated whether the Th2 cytokine interleukin-4 (IL-4) induces eotaxin production in human corneal epithelial cells and keratocytes. Primary cultures of human corneal epithelial cells and keratocytes were incubated with IL-4 and/or TNF-alpha for 48 h. Released eotaxin was measured by ELISA, and the eotaxin proteins were visualized by immunocytochemistry. Eotaxin mRNA expression in cultured cells was analyzed by RT-PCR. IL-4 induced eotaxin production in keratocytes in a dose- and time-dependent manner which was enhanced by TNF-alpha. There was no detectable eotaxin produced by corneal epithelial cells (<5 pg/ml). The cytoplasm of keratocytes incubated with IL-4 stained positively against anti-eotaxin antibodies, while eotaxin mRNA was detected in keratocytes incubated with IL-4. Human corneal keratocytes, but not epithelial cells, are capable of producing eotaxin by stimulation with IL-4. Our results suggest that eotaxin production in keratocytes induced by IL-4 may play an important role in eosinophil recruitment to corneal ulcers in allergic ocular disease. Eotaxin production by keratocytes may explain the severity of allergic disease involving the corneal stroma.


Physical Activity, Sedentary Behavior, and Physical Function in Older Adults With Multiple Sclerosis.
Older adults with multiple sclerosis (MS) experience the combined effects of aging and a chronic, disabling neurological disease on physical activity, sedentary behavior, and physical function. This study examined associations among light and moderate-to-vigorous physical activity (LPA and MVPA), sedentary behavior, and physical function in older adults with MS. Forty older adults with MS (median age = 60 years) who had a median Expanded Disability Status Scale score of 4.5 wore an accelerometer for a 7-day period and completed the Short Physical Performance Battery (SBBP), 6-minute walk (6MW), and timed 25-foot walk (T25FW). LPA was associated with SPPB (rs = .551, p < 0.01), 6MW (rs = .660, p < 0.01), and T25FW (rs = .623, p < 0.01) scores; MVPA was associated with 6MW (rs = .529, p < 0.01) and T25FW (rs = .403, p < 0.01) scores. There were significant associations between LPA, but not MVPA, with SPPB (β = .583, p < 0.01), 6MW (β = .613, p < 0.01), and T25FW (β = .627, p < 0.01) scores in linear regression analyses. Older adults with MS who engaged in more LPA demonstrated better physical function and therefore LPA might be a target of future behavioral interventions.


Molecular diversity of Epstein-Barr virus IgG and IgA antibody responses in nasopharyngeal carcinoma: a comparison of Indonesian, Chinese, and European subjects.
Epstein-Barr virus (EBV)-specific immunoblot analysis was used to reveal the molecular diversity of immunoglobulin (Ig) G and IgA antibody responses against Epstein-Barr nuclear antigen (EBNA), early antigen (EA), and viral capsid antigen (VCA) in serum samples from patients with nasopharyngeal carcinoma (NPC) and control subjects, by use of immunofluorescence assay (IFA). Control donors (n=150) showed IgG responses to few EBV proteins--VCA-p18, VCA-p40, EBNA1, and Zebra--and sporadically weak IgA reactivity to EBNA1 and VCA-p18. Patients with NPC stage 1 (n=6) had similar response patterns. Patients with NPC stage 2-4 (n=132) showed significantly more diverse IgG and IgA responses to EA and VCA proteins--VCA-p18/-p40, EBNA1, Z-encoded broadly reactive activator, and EAd-p47/54, -DNAse, -thymidine kinase, and -p138. No correlation was found between IFA titers and the number of EBV proteins recognized by IgG or IgA. Our results reveal dissimilarity between EBV polypeptides recognized by IgG and IgA antibodies, which suggests independent B cell triggering events.


Efficacy and Safety of UI05MSP015CT in Functional Dyspepsia: A Randomized, Controlled Trial.
To evaluate the efficacy and safety of a controlled release, once-daily formulation of mosapride (UI05MSP015CT) in patients with functional dyspepsia (FD). Patients with FD were randomly assigned (1:1) to receive either UI05MSP015CT (15 mg once a day, study group) or mosapride (5 mg three times a day, control group) and corresponding placebo for 4 weeks. The primary endpoint was a change in the gastrointestinal symptom score (GIS) evaluated at enrollment and after 4 weeks. Secondary endpoints were changes in the Nepean Dyspepsia Index-Korean version (NDI-K), rate of satisfactory symptom relief, and rate of adverse events. A total of 138 patients were enrolled (female, 73.9%; mean age, 44.0±15.4 years). After excluding patients who violated the study protocol, 59 and 58 patients from the study and control groups, respectively, were included in the per-protocol analysis. No difference was observed in drug compliance between the control and study groups (97.07%±4.52% vs 96.85%±6.05%, p=0.870). Changes in GIS scores were -9.69±6.44 and -10.01±5.92 in the study and control groups. The mean difference in GIS change between groups was 0.33 (95% confidence interval, -1.75 to 2.41), demonstrating non-inferiority of UI-05MSP015CT (p=0.755). The rate of satisfactory symptom relief was not different between the study and control groups (39.0% vs 56.9%, p=0.053). No differences in change in NDI-K score (14.3 vs 16.9, p=0.263) or rates of adverse events (12.9% vs. 4.4%, p=0.062) were observed between the study and control groups. Once-daily mosapride is not inferior to conventional mosapride in efficacy and is safe in patients with FD.


Refining perception-based farmer typologies with the analysis of past census data.
Perception-based typologies have been used to explore the decision making process of farmers and to inform policy design. These typologies have been criticised, however, for not fully capturing true farmer behaviour, and are consequently limited for supporting policy formulation. We present a method that develops a typology, using a social survey approach based on how farmers perceive their environment (e.g. birds and agri-environmental schemes). We then apply time-series census data on past farm strategies (i.e. land use allocation, management style and participation into agri-environmental schemes) to refine these typologies. Consequently, this offers an approach to improving the profiling of farmer types, and strengthens the validity of input into future agricultural policies. While the social survey highlights a certain degree of awareness towards birds with respect to farmer types, the analysis of past farm strategies indicated that farmers did not entirely follow their stated objectives. External factors such as input and output price signals and subsidy levels had a stronger influence on their strategies rather than stated environmental and social issues. Consequently, the refining of farmer types using this approach would aid the design of policy instruments, which integrate ecological issues within planning.


Awakening the Sleeping Carboxylase Function of Enzymes: Engineering the Natural CO2-Binding Potential of Reductases.
Developing new carbon dioxide (CO2) fixing enzymes is a prerequisite to create new biocatalysts for diverse applications in chemistry, biotechnology and synthetic biology. Here we used bioinformatics to identify a "sleeping carboxylase function" in the superfamily of medium-chain dehydrogenases/reductases (MDR), i.e. enzymes that possess a low carboxylation side activity next to their original enzyme reaction. We show that propionyl-CoA synthase from Erythrobacter sp. NAP1, as well as an acrylyl-CoA reductase from Nitrosopumilus maritimus possess carboxylation yields of 3 ± 1 and 4.5 ± 0.9%. We use rational design to engineer these enzymes further into carboxylases by increasing interactions of the proteins with CO2 and suppressing diffusion of water to the active site. The engineered carboxylases show improved CO2-binding and kinetic parameters comparable to naturally existing CO2-fixing enzymes. Our results provide a strategy to develop novel CO2-fixing enzymes and shed light on the emergence of natural carboxylases during evolution.


Quality of cellular attachment to various root-end filling materials.
This study investigated cellular attachment to 6 root-end filling materials as a measure of the biocompatibility of the materials. Class I retrograde cavities were prepared in root slices and these cavities were filled with the test materials, and incubated with Balb/C 3T3 fibroblasts for 24 h. Root slices with the cavities left empty served as the controls. The root slices were then processed for scanning electron microscopy, and were viewed to assess the quality of cellular attachment by observing the shape of cells, spread, and membrane outline. The best cellular attachment was observed at MTA and Geristore surfaces: cells exhibited characteristic elongated fibroblastic morphology, with projections of lamellipodia, filopodia, blebs, and microvilli from their surfaces, reflecting good attachment to the material. Fibroblasts attached poorly to the surfaces of IRM, Super EBA, KetacFil and Retroplast. Furthermore, the cells did not attach well to the tooth structure next to IRM and Super EBA. The present study demonstrated a variation in cellular attachment to different root-end filling materials with the best cellular attachment to the surfaces of MTA and Geristore. IRM and Super EBA, KetacFil and Retroplast rendered poor attachment.


Feral swine disturbance at important archaeological sites.
Feral swine are well known as environmentally destructive invasive animals in many areas around the world, where they degrade native habitats, harm rare plant and animal species, damage agricultural interests, and spread disease. We provide the first quantification of their potential as agents of disturbance at archaeological sites. Our study was conducted in south-central Florida at Avon Park Air Force Range, a base comprising over 40,000 ha and containing many archaeological sites. To determine the identifiable prevalence of feral swine disturbance, we examined 36 sites registered with the Florida State Historic Preservation Office and also eligible for inclusion in the National Register of Historic Places (NRHP). Moreover, we evaluated the extent of swine disturbance at a prehistoric site of extraordinary significance to Florida's prehistory, "Dead Cow." Fifteen of the 36 NRHP-eligible sites (42 %) had some level of swine disturbance, including 14 of 30 (47 %) sites known to have artifacts within 20 cm of the surface (well within swine rooting depths). At the Dead Cow site, we documented disturbance at 74 % of shovel test points. Sites with shallow artifact depositions appeared highly vulnerable to disturbance by feral swine, threatening destruction of artifact stratigraphy and provenience. Our observations likely are a minimal representation of accumulated damage. These irreplaceable sites tell the area's land use story across the millennia. That they are under threat from feral swine should serve broad notice of potential threats that feral swine may pose to archaeological sites globally, making effective swine management imperative for site protection.


Prospects for strongly coupled atom-photon quantum nodes.
We discuss the trapping of cold atoms within microscopic voids drilled perpendicularly through the axis of an optical waveguide. The dimensions of the voids considered are between 1 and 40 optical wavelengths. By simulating light transmission across the voids, we find that appropriate shaping of the voids can substantially reduce the associated loss of optical power. Our results demonstrate that the formation of an optical cavity around such a void could produce strong coupling between the atoms and the guided light. By bringing multiple atoms into a single void and exploiting collective enhancement, cooperativities ~400 or more should be achievable. The simulations are carried out using a finite difference time domain method. Methods for the production of such a void and the trapping of cold atoms within it are also discussed.