Mortality and life expectancy in relation to long-term cigarette, cigar and pipe smoking: the Zutphen Study.
To study the effect of long-term smoking on all-cause and cause-specific mortality, and to estimate the effects of cigarette and cigar or pipe smoking on life expectancy. A long-term prospective cohort study. Zutphen, The Netherlands. 1373 men from the Zutphen Study, born between 1900 and 1920 and studied between 1960 and 2000. Hazard ratios for the type of smoking, amount and duration of cigarette smoking, obtained from a time-dependent Cox regression model. Absolute health effects of smoking are expressed as differences in life expectancy and the number of disease-free years of life. Duration of cigarette smoking was strongly associated with mortality from cardiovascular disease, lung cancer and chronic obstructive pulmonary disease, whereas both the number of cigarettes smoked as well as duration of cigarette smoking were strongly associated with all-cause mortality. Average cigarette smoking reduced the total life expectancy by 6.8 years, whereas heavy cigarette smoking reduced the total life expectancy by 8.8 years. The number of total life-years lost due to cigar or pipe smoking was 4.7 years. Moreover, cigarette smoking reduced the number of disease-free life-years by 5.8 years, and cigar or pipe smoking by 5.2 years. Stopping cigarette smoking at age 40 increased the life expectancy by 4.6 years, while the number of disease-free life-years was increased by 3.0 years. Cigar or pipe smoking reduces life expectancy to a lesser extent than cigarette smoking. Both the number of cigarettes smoked and duration of smoking are strongly associated with mortality risk and the number of life-years lost. Stopping smoking after age 40 has major health benefits.


Expression of factor VIII in recombinant and transgenic systems.
Deficiency in a coagulation factor VIII (FVIII) causes a genetic disorder hemophilia A, which is treated by repeated infusions of expensive FVIII products. Recombinant FVIII (rFVIII), the culmination of years of extensive international research, is an important alternative to plasma-derived FVIII (pdFVIII) and is considered to have a higher margin of safety. Advances in biotechnology allowed production of rFVIII at industrial scale, which significantly improved treatment of hemophilia A patients. We review the contemporary methods used for FVIII expression in mammalian cell culture systems and discuss the factors responsible for insufficient recoveries of rFVIII, such as inefficient accumulation of FVIII mRNA in the cell, complexity of the mechanisms of FVIII secretion, and instability of secreted FVIII. The approaches to improve the yield of rFVIII in cell culture systems include genetic engineering of B-domain-deleted FVIII, introduction of introns into FVIII cDNA constructs for more efficient processing and accumulation of FVIII mRNA, and introduction of mutations into chaperone-binding sites of FVIII to improve its secretion. Design of FVIII with prolonged half-life in vivo is considered as another promising direction in improving rFVIII protein and efficiency of hemophilia A therapy. As an alternative to expression of rFVIII in cell culture systems, we discuss production of rFVIII in transgenic animals, where high levels of rFVIII have been successfully secreted into milk. We also pay attention to the major limitations of this approach, such as safety issues associated with potential transmission of animal pathogens. Finally, we present a brief characterization of commercial recombinant FVIII products currently available on the market for hemophilia A treatment.


Regional and remote occupational therapy: a preliminary exploration of private occupational therapy practice.
Private providers of Medicare funded services are an integral part of the Australian primary health-care system. Evidence on private occupational therapy practice in rural and remote Australian settings is sparse. Providers of private occupational therapy services in outer regional and remote areas were surveyed regarding location and type of services provided, practice models and demographics. Descriptive statistics were used to summarise the findings. Thirty-seven respondents completed the survey. In remote areas a number of occupational therapy services are not provided and no practices are based in very remote towns. One quarter of respondents visit at least five towns each week and one third had other paid employment. Nearly half indicated they will leave private practice within the next five years and nearly one third believed they could not sustain their practice. Medicare Chronic Disease Management was the main income source of only around half the respondents. There is a potential market failure of private occupational therapy in rural and remote areas, impacting on people who need these services and on workforce replacement. Contributing factors include population imbalance between cities and regional/remote areas, financial implications where only face-to-face contact is paid for and low incomes and levels of health insurance in regional/remote areas. Potential strategies include addressing the lack of reimbursement for travel, enabling private providers to overcome barriers to providing student placements and recognising rural practice as a specialist field.


The Listeria transcriptional landscape from saprophytism to virulence.
The bacterium Listeria monocytogenes is ubiquitous in the environment and can lead to severe food-borne infections. It has recently emerged as a multifaceted model in pathogenesis. However, how this bacterium switches from a saprophyte to a pathogen is largely unknown. Here, using tiling arrays and RNAs from wild-type and mutant bacteria grown in vitro, ex vivo and in vivo, we have analysed the transcription of its entire genome. We provide the complete Listeria operon map and have uncovered far more diverse types of RNAs than expected: in addition to 50 small RNAs (<500 nucleotides), at least two of which are involved in virulence in mice, we have identified antisense RNAs covering several open-reading frames and long overlapping 5' and 3' untranslated regions. We discovered that riboswitches can act as terminators for upstream genes. When Listeria reaches the host intestinal lumen, an extensive transcriptional reshaping occurs with a SigB-mediated activation of virulence genes. In contrast, in the blood, PrfA controls transcription of virulence genes. Remarkably, several non-coding RNAs absent in the non-pathogenic species Listeria innocua exhibit the same expression patterns as the virulence genes. Together, our data unravel successive and coordinated global transcriptional changes during infection and point to previously unknown regulatory mechanisms in bacteria.


Coupling dark metabolism to electricity generation using photosynthetic cocultures.
We investigated the role of green sulfur bacteria inlight-responsive electricity generation in microbial electrochemical cells (MXCs). We operated MXCs containing either monocultures or defined cocultures of previously enriched phototrophic Chlorobium and anode-respiring Geobacter under anaerobic conditions in the absence of electron donor. Monoculture control MXCs containing Geobacter or Chlorobium neither responded to light nor produced current, respectively. Instead, light-responsive current generation occurred only in coculture MXCs. Current increased above background levels only in the dark and declined slowly over 96 h. This pattern suggested that Chlorobium exhausted intracellular glycogen reserves via dark fermentation to supply an electron donor, presumably acetate, to Geobacter. With medium containing sulfide as the sole photosynthetic electron donor, current generation had a similar and reproducible negative light response. To investigate whether this metabolic interaction also occurred without an electrode, we performed coculture experiments in batch serum bottles. In this setup, sulfide served as the sole electron donor, whose oxidation by Chlorobium was required to provide S(0) as the electron acceptor to Geobacter. Copies of Geobacter 16S rDNA increased approximately 14-fold in batch bottle cocultures containing sulfide compared to those lacking sulfide, and did not decline after termination of sulfide feeding. These results suggest that products of both photosynthesis and dark fermentation by Chlorobium were sufficient both to yield an electrochemical response by Geobacter biofilms, and to promote Geobacter growthin batch cocultures. Our work expands upon the fusion of MXCs with coculture techniques and reinforces the utility of microbial electrochemistry for sensitive, real-time monitoring of microbial interactions in which a metabolic intermediate can be converted to electrical current.


Isolation of cells for selective treatment and analysis using a magnetic microfluidic chip.
This study describes the development and testing of a magnetic microfluidic chip (MMC) for trapping and isolating cells tagged with superparamagnetic beads (SPBs) in a microfluidic environment for selective treatment and analysis. The trapping and isolation are done in two separate steps; first, the trapping of the tagged cells in a main channel is achieved by soft ferromagnetic disks and second, the transportation of the cells into side chambers for isolation is executed by tapered conductive paths made of Gold (Au). Numerical simulations were performed to analyze the magnetic flux and force distributions of the disks and conducting paths, for trapping and transporting SPBs. The MMC was fabricated using standard microfabrication processes. Experiments were performed with E. coli (K12 strand) tagged with 2.8 μm SPBs. The results showed that E. coli can be separated from a sample solution by trapping them at the disk sites, and then isolated into chambers by transporting them along the tapered conducting paths. Once the E. coli was trapped inside the side chambers, two selective treatments were performed. In one chamber, a solution with minimal nutrition content was added and, in another chamber, a solution with essential nutrition was added. The results showed that the growth of bacteria cultured in the second chamber containing nutrient was significantly higher, demonstrating that the E. coli was not affected by the magnetically driven transportation and the feasibility of performing different treatments on selectively isolated cells on a single microfluidic platform.


Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11.
The purpose of the research was to study the purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. The enzyme was purified in a 2-step procedure involving ammonium sulfate precipitation and Sephadex G-200 gel permeation chromatography. The enzyme was shown to have a relative low molecular weight of 15 kd by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and was purified 21-fold with a yield of 7.5%. It was most active at 60 degrees C, pH 10, with casein as substrate. It was stable between pH 8 and 10. This enzyme was almost 100% stable at 60 degrees C even after 350 minutes of incubation. It was strongly activated by metal ions such as Ca+2, Mg+2, and Mn+2. Enzyme activity was inhibited strongly by phenylmethyl sulphonyl fluoride (PMSF) and diisopropyl fluorophosphates (DFP) but was not inhibited by ethylene diamine tetra acetic acid (EDTA), while a slight inhibition was observed with iodoacetate, p-chloromercuric benzoate (pCMB), and beta-mercaptoethanol (beta-ME). The compatibility of the enzyme was studied with commercial and local detergents in the presence of 10mM CaCl2 and 1M glycine. The addition of 10mM CaCl2 and 1M glycine, individually and in combination, was found to be very effective in improving the enzyme stability where it retained 52% activity even after 3 hours. This enzyme improved the cleansing power of various detergents. It removed blood stains completely when used with detergents in the presence of 10mM CaCl2 and 1M glycine.


The estimation of body height from ulna length in healthy adults from different ethnic groups.
Assessments of nutritional status frequently incorporate a measure of height to evaluate a person's relative thinness or fatness. Because height is often difficult to quantify, it may be predicted from alternative anthropometric measurements, including ulna length. Little information is available about the accuracy of these predictions in an ethnically diverse population. The present study aimed to evaluate published equations for predicting height from ulna length in adults from different ethnic groups. Ulna length and standing height were measured in a gender-stratified sample of 60 Asian, 69 Black and 65 White healthy volunteers, aged 21-65 years. Height was predicted from ulna length using the Malnutrition Universal Screening Tool (MUST) equations and compared against the measured values. Linear regression analysis was used to develop equations to estimate height from ulna length and to explore the relationship between height and ulna length in subgroups. The mean (SD) age for Asian, Black and White in men was 31.7 (11.0), 32.0 (10.3) and 38.6 (12.5) years and in women was 26.2 (5.4), 32.6 (8.9) and 35.7 (11.7); the mean (SD) height in men was 170.9 (5.2), 178.1 (7.3) and 176.3 (7.7) cm and in women was 157.7 (4.7), 164.0 (5.9) and 163.7 (6.2) cm. Ulna length and measured height were significantly correlated among all subgroups, except Asian women (r=0.11, P=0.57). The mean (SD) difference between predicted and measured height showed significant overestimates for Asian and Black men [4.0 (4.8) and 6.7 (5.3) cm] and Asian and Black women [6.4 (4.9) and 4.4 (4.9) cm] but not for White men and women. The MUST equations for predicting height from ulna length in healthy adults should be used with some caution among ethnically diverse populations, particularly in Asian women.


Refined Data Analysis Provides Clinical Evidence for Central Nervous System Control of Chronic Glaucomatous Neurodegeneration.
Refined data analysis was performed to assess binocular visual field conservation in patients with bilateral glaucomatous damage to determine whether unilateral visual field loss is random, anatomically symmetric, or nonrandom in relation to the fellow eye. This was a case-control study of 47 consecutive patients with bilaterally severe glaucoma; each right eye visual field locus was paired with randomly selected coisopteric left eye loci, with 760,000 (10,000 complete sets of 76 loci) such iterations performed per subject. The potential role of anatomic symmetry in bilateral visual field conservation was also assessed by pairing mirror-image loci of the paired fields. The mean values of the random coisopteric and the symmetric mirror pairings were compared with natural point-for-point pairings of the two eyes by paired t-test. Mean unilateral thresholds across the entire visual field were 18.9 dB left and 19.9 dB right (average 19.4), 4 dB lower than the better of the naturally paired concomitant loci of 23.4 dB (P < 10-15). A remarkable natural tendency for conservation of the binocular visual field was confirmed, far stronger than explicable by random chance or anatomic symmetry (P < 0.0001), and reaffirmed by subsequent prospective simultaneous binocular visual field retesting of an arbitrary subset (n = 16) of the study population (P < 0.0001). Refined data analysis of paired visual fields confirms the existence of a natural optimization of binocular visual function in severe bilateral glaucoma via interlocking fields that could be created only by central nervous system (CNS) involvement. Integrated bilateral visual field analysis should better define actual visual disability and more accurately reflect the functional efficacy of current ocular and future CNS-oriented therapeutic approaches to the treatment of glaucoma. Glaucomatous eyes provide a highly accessible paired-organ study model for developing therapeutics to optimize conservation of function in neurodegenerative disorders.


Identification and characterization of two novel brain-derived immunoglobulin superfamily members with a unique structural organization.
We recently used a differential display PCR screen to identify secreted and transmembrane proteins that are highly expressed in the developing rat basilar pons, a prominent ventral hindbrain nucleus used as a model for studies of neuronal migration, axon outgrowth, and axon-target recognition. Here we describe cloning and characterization of one of these molecules, now called MDGA1, and a closely related homologue, MDGA2. Analyses of the full-length coding region of MDGA1 and MDGA2 indicate that they encode proteins that comprise a novel subgroup of the Ig superfamily and have a unique structural organization consisting of six immunoglobulin (Ig)-like domains followed by a single MAM domain. Biochemical characterization demonstrates that MDGA1 and MDGA2 proteins are highly glycosylated, and that MDGA1 is tethered to the cell membrane by a GPI anchor. The MDGAs are differentially expressed by subpopulations of neurons in both the central and peripheral nervous systems, including neurons of the basilar pons, inferior olive, cerebellum, cerebral cortex, olfactory bulb, spinal cord, and dorsal root and trigeminal ganglia. Little or no MDGA expression is detected outside of the nervous system of developing rats. The similarity of MDGAs to other Ig-containing molecules and their temporal-spatial patterns of expression within restricted neuronal populations, for example migrating pontine neurons and D1 spinal interneurons, suggest a role for these novel proteins in regulating neuronal migration, as well as other aspects of neural development, including axon guidance.


Optical-CT 3D Dosimetry Using Fresnel Lenses with Minimal Refractive-Index Matching Fluid.
Telecentric optical computed tomography (optical-CT) is a state-of-the-art method for visualizing and quantifying 3-dimensional dose distributions in radiochromic dosimeters. In this work a prototype telecentric system (DFOS-Duke Fresnel Optical-CT Scanner) is evaluated which incorporates two substantial design changes: the use of Fresnel lenses (reducing lens costs from $10-30K t0 $1-3K) and the use of a 'solid tank' (which reduces noise, and the volume of refractively matched fluid from 1 ltr to 10 cc). The efficacy of DFOS was evaluated by direct comparison against commissioned scanners in our lab. Measured dose distributions from all systems were compared against the predicted dose distributions from a commissioned treatment planning system (TPS). Three treatment plans were investigated including a simple four-field box treatment, a multiple small field delivery, and a complex IMRT treatment. Dosimeters were imaged within 2 h post irradiation, using consistent scanning techniques (360 projections acquired at 1 degree intervals, reconstruction at 2mm). DFOS efficacy was evaluated through inspection of dose line-profiles, and 2D and 3D dose and gamma maps. DFOS/TPS gamma pass rates with 3%/3mm dose difference/distance-to-agreement criteria ranged from 89.3% to 92.2%, compared to from 95.6% to 99.0% obtained with the commissioned system. The 3D gamma pass rate between the commissioned system and DFOS was 98.2%. The typical noise rates in DFOS reconstructions were up to 3%, compared to under 2% for the commissioned system. In conclusion, while the introduction of a solid tank proved advantageous with regards to cost and convenience, further work is required to improve the image quality and dose reconstruction accuracy of the new DFOS optical-CT system.


Behavioural assessment of the effects of tumour growth in rats and the influence of the analgesics carprofen and meloxicam.
Very little is known concerning the occurrence of pain in cancer research models. We wished to establish whether a behaviour-based approach, originally developed to assess postoperative pain, could be used to determine positive effects of the analgesics carprofen and meloxicam in rats that might be experiencing pain during tumour development in an orthotopic model of bladder cancer. An invasive but non-metastatic rat bladder cancer cell line was surgically implanted into the bladder wall of 57 inbred Fisher344 rats. The rats underwent daily clinical assessments. When clinical signs consistent with chronic pain were apparent, behavioural data were collected from 44 animals during 2 x 10 min periods, immediately before and one hour after a subcutaneous injection of either physiological saline (0.9%; 0.2 ml/100 g), carprofen (5 mg/kg) or meloxicam (2 mg/kg). Treatment-associated behaviour changes were then compared between groups. The lack of active behaviour, both before and after each treatment, was consistent with established clinical signs of pain. The rats were so inactive following the treatment that the behavioural technique we had previously developed was of comparatively little use in determining either pain severity or analgesic efficacy. One very prominent effect, however, was an increase in ventral abdominal licking in the control (saline) group. As this was absent in rats given meloxicam or carprofen, and has previously been considered to indicate pain emanating from damaged tissue, it was concluded that the analgesic-treated rats gained at least some benefit from the drug treatments, but it was not possible to gauge the extent of this. Handling for examination or treatment may have intensified pain in rats in the control group, and so this should be avoided whenever possible. It is likely that post-surgical pain differs markedly from cancer pain, so a different set of behavioural markers may be needed to assess it effectively. More intensive behaviour monitoring may help to develop a suitable technique for detecting the onset of, and assess the severity of pain that may occur during tumour development.


Global Biobank Engine: enabling genotype-phenotype browsing for biobank summary statistics.
Large biobanks linking phenotype to genotype have led to an explosion of genetic association studies across a wide range of phenotypes. Sharing the knowledge generated by these resources with the scientific community remains a challenge due to patient privacy and the vast amount of data. Here, we present Global Biobank Engine (GBE), a web-based tool that enables exploration of the relationship between genotype and phenotype in biobank cohorts, such as the UK Biobank. GBE supports browsing for results from genome-wide association studies, phenome-wide association studies, gene-based tests and genetic correlation between phenotypes. We envision GBE as a platform that facilitates the dissemination of summary statistics from biobanks to the scientific and clinical communities. GBE currently hosts data from the UK Biobank and can be found freely available at biobankengine.stanford.edu.


Obsessive-compulsive scale of the child behavior checklist: specificity, sensitivity, and predictive power.
To create an obsessive-compulsive disorder subscale (OCS) of the Child Behavior Checklist (CBCL) and to determine its internal consistency, sensitivity, specificity, and positive and negative predictive power to identify obsessive-compulsive disorder (OCD) in children and adolescents. Three samples of equal size (n = 73) of children and adolescents, matched for age, gender, and race, were selected for these analyses: 1) a clinically ascertained OCD group, 2) a psychiatrically treated group whose records revealed no evidence of OCD, and 3) a general population control group. An OCS was created by applying factor analysis to 11 CBCL items. Examinations of internal consistency, sensitivity, specificity, and positive and negative predictive value were undertaken. Of 11 items hypothesized to predict OCD, 8 items were retained after factor analyses (smallest factor loading: 0.49) and used to calculate OCS scores. The retained items displayed excellent internal consistency (Cronbach's alpha coefficient = 0.84). OCD participants had significantly higher OCS scores than either psychiatrically treated or general population control groups. With the use of the 2 cutoff scores closest to the true rate of OCD in the overall sample, sensitivity was 75.3% to 84.9%, specificity was 82.2% to 92.5%, positive predictive value was 70.5% to 83.3%, and negative predictive value was 88.2% to 91.6%. The performance of the proposed CBCL OCS compares favorably with that of the only previously studied screening instrument for OCD, the Leyton Obsessional Inventory-Child Version. Unlike the Leyton Obsessional Inventory-Child Version, the CBCL is already in widespread use as a screen for most other forms of psychopathology. As the performance of the CBCL OCS will need to be replicated in other sample populations, data with various cutoff levels are provided to enable investigators and clinicians to tailor its use to specific study populations.


Development of a Generic Physiologically-Based Pharmacokinetic Model for Lactation and Prediction of Maternal and Infant Exposure to Ondansetron via Breast Milk.
Ondansetron is commonly used in breastfeeding mothers to treat nausea and vomiting. There is limited information in humans regarding safety of ondansetron exposure to nursing infants and no adequate study looking at ondansetron pharmacokinetics during lactation. We developed a generic physiologically-based pharmacokinetic lactation model for small molecule drugs and applied this model to predict ondansetron transfer into breast milk and characterize infant exposure. Drug-specific model inputs were parameterized using data from the literature. Population-specific inputs were derived from a previously conducted systematic literature review of anatomic and physiologic changes in postpartum women. Model predictions were evaluated using ondansetron plasma and breast milk concentration data collected prospectively from 78 women in the Commonly Used Drugs During Lactation and infant Exposure (CUDDLE) study. The final model predicted breast milk and plasma exposures following a single 4 mg dose of intravenous ondansetron in 1,000 simulated women who were 2 days postpartum. Model predictions showed good agreement with observed data. Breast milk median prediction error (MPE) was 18.4% and median absolute prediction error (MAPE) was 53.0%. Plasma MPE was 32.5% and MAPE was 43.2%. The model-predicted daily and relative infant doses were 0.005 mg/kg/day and 3.0%, respectively. This model adequately predicted ondansetron passage into breast milk. The calculated low relative infant dose indicates that mothers receiving ondansetron can safely breastfeed. The model building blocks and population database are open-source and can be adapted to other drugs.


Relating inter-individual differences in metacognitive performance on different perceptual tasks.
Human behavior depends on the ability to effectively introspect about our performance. For simple perceptual decisions, this introspective or metacognitive ability varies substantially across individuals and is correlated with the structure of focal areas in prefrontal cortex. This raises the possibility that the ability to introspect about different perceptual decisions might be mediated by a common cognitive process. To test this hypothesis, we examined whether inter-individual differences in metacognitive ability were correlated across two different perceptual tasks where individuals made judgments about different and unrelated visual stimulus properties. We found that inter-individual differences were strongly correlated between the two tasks for metacognitive ability but not objective performance. Such stability of an individual's metacognitive ability across different perceptual tasks indicates a general mechanism supporting metacognition independent of the specific task.


A Stimulator ASIC Featuring Versatile Management for Vestibular Prostheses.
This paper presents a multichannel stimulator ASIC for an implantable vestibular prosthesis. The system features versatile stimulation management which allows fine setting of the parameters for biphasic stimulation pulses. To address the problem of charge imbalance due to rounding errors, the digital processor can calculate and provide accurate charge correction. A technique to reduce the data rate to the stimulator is described. The stimulator ASIC was implemented in 0.6-μ m high-voltage CMOS technology occupying an area of 2.27 mm(2). The measured performance of the ASIC has been verified using vestibular electrodes in saline.


Secreted α-Klotho maintains cartilage tissue homeostasis by repressing NOS2 and ZIP8-MMP13 catabolic axis.
Progressive loss of tissue homeostasis is a hallmark of numerous age-related pathologies, including osteoarthritis (OA). Accumulation of senescent chondrocytes in joints contributes to the age-dependent cartilage loss of functions through the production of hypertrophy-associated catabolic matrix-remodeling enzymes and pro-inflammatory cytokines. Here, we evaluated the effects of the secreted variant of the anti-aging hormone α-Klotho on cartilage homeostasis during both cartilage formation and OA development. First, we found that α-Klotho expression was detected during mouse limb development, and transiently expressed during in vitro chondrogenic differentiation of bone marrow-derived mesenchymal stem cells. Genome-wide gene array analysis of chondrocytes from OA patients revealed that incubation with recombinant secreted α-Klotho repressed expression of the NOS2 and ZIP8/MMP13 catabolic remodeling axis. Accordingly, α-Klotho expression was reduced in chronically IL1β-treated chondrocytes and in cartilage of an OA mouse model. Finally, in vivo intra-articular secreted α-Kotho gene transfer delays cartilage degradation in the OA mouse model. Altogether, our results reveal a new tissue homeostatic function for this anti-aging hormone in protecting against OA onset and progression.


Anesthetics rapidly promote synaptogenesis during a critical period of brain development.
Experience-driven activity plays an essential role in the development of brain circuitry during critical periods of early postnatal life, a process that depends upon a dynamic balance between excitatory and inhibitory signals. Since general anesthetics are powerful pharmacological modulators of neuronal activity, an important question is whether and how these drugs can affect the development of synaptic networks. To address this issue, we examined here the impact of anesthetics on synapse growth and dynamics. We show that exposure of young rodents to anesthetics that either enhance GABAergic inhibition or block NMDA receptors rapidly induce a significant increase in dendritic spine density in the somatosensory cortex and hippocampus. This effect is developmentally regulated; it is transient but lasts for several days and is also reproduced by selective antagonists of excitatory receptors. Analyses of spine dynamics in hippocampal slice cultures reveals that this effect is mediated through an increased rate of protrusions formation, a better stabilization of newly formed spines, and leads to the formation of functional synapses. Altogether, these findings point to anesthesia as an important modulator of spine dynamics in the developing brain and suggest the existence of a homeostatic process regulating spine formation as a function of neural activity. Importantly, they also raise concern about the potential impact of these drugs on human practice, when applied during critical periods of development in infants.


Molecular layers in thin supported films exhibit the same scaling as the bulk between slow relaxation and vibrational dynamics.
We perform molecular-dynamics simulations of a supported molecular thin film. By varying thickness and temperature, we observe anisotropic mobility as well as strong gradients of both the vibrational motion and the structural relaxation through film layers with monomer-size thickness. We show that the gradients of the fast and the slow dynamics across the layers (except the adherent layer to the substrate) comply, without any adjustment, with the same scaling between the structural relaxation time and the Debye-Waller factor originally observed in the bulk [Larini et al., Nat. Phys., 2008, 4, 42]. The scaling is not observed if the average dynamics of the film is inspected. Our results suggest that the solidification process of each layer may be tracked by knowing solely the vibrational properties of the layer and the bulk.


Oxidation of iodide and iodine on birnessite (delta-MnO2) in the pH range 4-8.
The oxidation of iodide by synthetic birnessite (delta-MnO(2)) was studied in perchlorate media in the pH range 4-8. Iodine (I(2)) was detected as an oxidation product that was subsequently further oxidized to iodate (IO(3)(-)). The third order rate constants, second order on iodide and first order on manganese oxide, determined by extraction of iodine in benzene decreased with increasing pH (6.3-7.5) from 1790 to 3.1M(-2) s(-1). Both iodine and iodate were found to adsorb significantly on birnessite with an adsorption capacity of 12.7 microM/g for iodate at pH 5.7. The rate of iodine oxidation by birnessite decreased with increasing ionic strength, which resulted in a lower rate of iodate formation. The production of iodine in iodide-containing waters in contact with manganese oxides may result in the formation of undesired iodinated organic compounds (taste and odor, toxicity) in natural and technical systems. The probability of the formation of such compounds is highest in the pH range 5-7.5. For pH <5 iodine is quickly oxidized to iodate, a non-toxic and stable sink for iodine. At pH >7.5, iodide is not oxidized to a significant extent.


A position statement on mental health in the post-2015 development agenda.
The Millennium Development Goals have guided development co-operation in the 15 years up to 2015, achieving some significant progress in the priorities on which they focused. As the framework for the post-2015 development agenda is negotiated, this article reviews the evidence for the place of mental health in broader development issues that have already been outlined in the run-up to 2015. If mental health is going to be recognised as having an essential role in development, there needs to be a consensus on priorities for advocacy. Various key issues emerged from a survey of stakeholders in the Movement for Global Mental Health (MGMH), leading to a Position Statement, which is now available for use by advocates. The priorities that emerged were increasing access to mental health services, and addressing human rights abuse, stigma, and exclusion. Mental health is a cross-cutting issue, and including it in frameworks for action will increase the likelihood of achieving global priorities for development such as poverty reduction, economic development, improved health, and ensuring the most vulnerable in society are not left behind.


Clinical Management of Pneumonitis in Patients Receiving Anti-PD-1/PD-L1 Therapy.
CASE STUDY A 48-year-old gentleman with metastatic melanoma currently receiving the cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, ipilimumab (Yervoy), and the programmed cell death protein 1 (PD-1) inhibitor, nivolumab (Opdivo), returned for evaluation prior to receiving cycle 2. The patient presented with new onset dyspnea and a non-productive cough over the past week, with a temperature of 100.6°F at home on one occasion. He was placed on observation for fever, cough, and shortness of breath. The patient had no previous history of lung disease and was a nonsmoker. Diminished breath sounds were noted on auscultation. However, the patient was without fever or chills, with a heart rate of 101 beats per minute and a blood pressure of 110/75 mm Hg. We obtained a computed tomography (CT) of his chest. The CT demonstrated diffuse ground-glass opacities in his bilateral lower lobes and some minor interstitial thickening of his right middle lobe, possibly suggestive of inflammation or cryptogenic organizing pneumonia. Based on his presentation and CT findings, the patient was initially treated empirically with antibiotics for suspected pneumonia vs. pneumonitis. During the first 12 hours in observation, the patient experienced increasing dyspnea and cough and was admitted to the hospital. Nebulizer treatments were administered with no improvement, so the patient was started on high-dose corticosteroids at 1 mg/kg, and pulmonary and infectious disease consults were ordered. After the administration of steroids, the patient's cough and breathing improved and he remained afebrile, eliciting a high suspicion for immune-related pneumonitis. The patient then underwent bronchoscopy to rule out other etiologies. Bronchoalveolar lavage was performed, which yielded no pathogenic organisms. The patient was placed on a 3-week course of a high-dose steroid taper, following which immunotherapy was reinstated. Within 4 days he again presented with similar symptoms, was restarted on high-dose steroids, and immunotherapy was permanently discontinued.


Protocol for studying cough frequency in people with pulmonary tuberculosis.
Cough is a key symptom of tuberculosis (TB) as well as the main cause of transmission. However, a recent literature review found that cough frequency (number of coughs per hour) in patients with TB has only been studied once, in 1969. The main aim of this study is to describe cough frequency patterns before and after the start of TB treatment and to determine baseline factors that affect cough frequency in these patients. Secondarily, we will evaluate the correlation between cough frequency and TB microbiological resolution. This study will select participants with culture confirmed TB from 2 tertiary hospitals in Lima, Peru. We estimated that a sample size of 107 patients was sufficient to detect clinically significant changes in cough frequency. Participants will initially be evaluated through questionnaires, radiology, microscopic observation drug susceptibility broth TB-culture, auramine smear microscopy and cough recordings. This cohort will be followed for the initial 60 days of anti-TB treatment, and throughout the study several microbiological samples as well as 24 h recordings will be collected. We will describe the variability of cough episodes and determine its association with baseline laboratory parameters of pulmonary TB. In addition, we will analyse the reduction of cough frequency in predicting TB cure, adjusted for potential confounders. Ethical approval has been obtained from the ethics committees at each participating hospital in Lima, Peru, Asociación Benéfica PRISMA in Lima, Peru, the Universidad Peruana Cayetano Heredia in Lima, Peru and Johns Hopkins University in Baltimore, USA. We aim to publish and disseminate our findings in peer-reviewed journals. We also expect to create and maintain an online repository for TB cough sounds as well as the statistical analysis employed.


Dual Encoder Attention U-net for Nuclei Segmentation.
Nuclei segmentation in whole slide images (WSIs) stained with Hematoxylin and Eosin (H&E) dye, is a key step in computational pathology which aims to automate the laborious process of manual counting and segmentation. Nuclei segmentation is a challenging problem that involves challenges such as touching nuclei resolution, small-sized nuclei, size, and shape variations. With the advent of deep learning, convolution neural networks (CNNs) have shown a powerful ability to extract effective representations from microscopic H&E images. We propose a novel dual encoder Attention U-net (DEAU) deep learning architecture and pseudo hard attention gating mechanism, to enhance the attention to target instances. We added a new secondary encoder to the attention U-net to capture the best attention for a given input. Since H captures nuclei information, we propose a stain-separated H channel as input to the secondary encoder. The role of the secondary encoder is to transform attention prior to different spatial resolutions while learning significant attention information. The proposed DEAU performance was evaluated on three publicly available H&E data sets for nuclei segmentation from different research groups. Experimental results show that our approach outperforms other attention-based approaches for nuclei segmentation.


Living safely with epilepsy: a key learning review.
This review focusses on counselling to reduce the risks of seizures in epilepsy patients. Risk reduction, whilst maximising the independence of persons with epilepsy, is a core element of epilepsy practice. The importance of the issue is reflected by the inclusion of risk assessment and reduction in the recent International League Against Epilepsy Curriculum for epileptology. This article addresses the key elements of epilepsy risk and provides practical guidance for counselling and risk reduction. A review of key publications was performed focusing on: (1) the risk of injury associated with seizures; (2) safety issues related to seizures and lifestyle; (3) SUDEP risk and avoidance; and (4) assessing individual risks. Risk is common in epilepsy and is multifactorial. Clinicians should be aware of the risk factors associated with different epilepsy features, such as seizure type, as well as those related to lifestyle and common indoor and outdoor activities. In addition, transcultural differences should be considered in risk assessment and counselling, e.g. regarding SUDEP or even driving, for which regulations vary across nations. Assessment of individual risk is evolving through a better understanding of risk factors and methodologies which can improve communication and empower patients to help identify and manage their individual risks.


Association of the gene polymorphisms of platelet glycoprotein Ia and IIb/IIIa with myocardial infarction and extent of coronary artery disease in the Korean population.
Platelet membrane receptor glycoproteins (GP) are essential for the platelet activation process, and the genetic polymorphisms in the genes that encode platelet glycoproteins have been proposed to influence the risk of acute coronary syndrome and atherosclerosis. In this study, we investigated the role of GPIa, HPA-1 and HPA-3 polymorphisms as putative risk factors for myocardial infarction (MI) and the extent of coronary artery disease. We selected 1,073 subjects who underwent coronary angiography; 242 had normal or minimal coronary atherosclerosis, and 831 patients had significant coronary artery disease (CAD). The genotype was determined by the methods of single base extension for C807T/G873A polymorphisms of GPIa, and restriction fragment length polymorphism for HPA-1 and HPA-3. The C807T and G873A polymorphisms of GPIa showed complete linkage in the Korean population. For HPA-1 gene polymorphism, only the HPA-1a/a (PlA1/A1) genotype was observed in 192 selected subjects from our study population. The distribution of GPIa (C807T/G873A) and HPA-3 genotypes did not differ significantly between normal subjects and CAD subjects. No significant association between MI and both gene polymorphisms was present. However, for the subgroup analysis of young male patients whose age was less than 56 years, the genotype frequency of HPA-3b/b was significantly lower in patients with MI compared to patients without a history of MI (7.5% vs. 20.0%, p=0.04). The odds ratio for HPA-3 b homozygosity versus the HPA-3a carrier was 0.32 (95% CI, 0.10- 0.99, p=0.04). Conclusively, HPA-3 polymorphism was associated with MI in Korean individuals younger than 56 years of age, but other polymorphisms of GP, which we studied, were not associated with both the extent of coronary atherosclerosis or MI.


HDAC Inhibition Upregulates PD-1 Ligands in Melanoma and Augments Immunotherapy with PD-1 Blockade.
Expression of PD-1 ligands by tumors and interaction with PD-1-expressing T cells in the tumor microenvironment can result in tolerance. Therapies targeting this coinhibitory axis have proven clinically successful in the treatment of metastatic melanoma, non-small cell lung cancer, and other malignancies. Therapeutic agents targeting the epigenetic regulatory family of histone deacetylases (HDAC) have shown clinical success in the treatment of some hematologic malignancies. Beyond direct tumor cell cytotoxicity, HDAC inhibitors have also been shown to alter the immunogenicity and enhance antitumor immune responses. Here, we show that class I HDAC inhibitors upregulated the expression of PD-L1 and, to a lesser degree, PD-L2 in melanomas. Evaluation of human and murine cell lines and patient tumors treated with a variety of HDAC inhibitors in vitro displayed upregulation of these ligands. This upregulation was robust and durable, with enhanced expression lasting past 96 hours. These results were validated in vivo in a B16F10 syngeneic murine model. Mechanistically, HDAC inhibitor treatment resulted in rapid upregulation of histone acetylation of the PD-L1 gene leading to enhanced and durable gene expression. The efficacy of combining HDAC inhibition with PD-1 blockade for treatment of melanoma was also explored in a murine B16F10 model. Mice receiving combination therapy had a slower tumor progression and increased survival compared with control and single-agent treatments. These results highlight the ability of epigenetic modifiers to augment immunotherapies, providing a rationale for combining HDAC inhibitors with PD-1 blockade.


President Kennedy's death: a poison arrow-assisted homicide.
"President John F. Kennedy's death was a neurotoxin-assisted homicide" is the hypothesis of this study. A review of medical evidence demonstrates evidence of a neurotoxin-assisted homicide. The convergence of three independent actions, or the signature traits of a neurotoxin-assisted homicide- the emergence of neurological signs consistent with a neurotoxin-induced paralysis, the induction of a small neck wound consistent with a flechette-transported neurotoxin entry wound, and the execution of a coverup to eliminate neurotoxin evidence, supports this hypothesis. This review suggests, JFK's death had all the signature traits of a neurotoxin-assisted homicide.


Post-traumatic stress and self-disclosure.
Post-traumatic stress (PTS) is a significant clinical problem in the general population. However, only a portion of those exposed to trauma develop PTS. Patterns of emotional self-disclosure have the potential to explain some of the individual differences in the development and continuation of symptoms. In this study, the authors investigated the links between emotional self-disclosure, as measured by the Emotional Self-Disclosure Scale (ESDS; W. E. Snell, R. S. Miller, & S. S. Belk, 1988). and a post-trauma psychological state, as measured by the Trauma Symptom Inventory (TSI; J. Briere, 1995). Their results showed that, in general, men engaged in less emotional self-disclosure than did women, and as TSI scores increased, the men were significantly less willing to disclose emotions of happiness. For women, as TSI scores increased they were significantly more willing to engage in talk about emotions related to anxiety but less willing to talk about emotions related to fear. The authors considered these data within current understandings of the role of emotional self-disclosure in the processing of traumatic experiences.


Impact of Chlamydia pneumoniae infection on survival rate after heart transplantation.
As Chlamydia pneumoniae (Cp), a common cause of respiratory infection, is of vasotropic character, chronic infection may be associated with the development of coronary disease, although there have been few reports on the impact of Cp infection on the post-orthotopic heart transplantation (OHT) survival rate. A total of 41 patients (4 females) were followed up for one year after OHT. Serology investigations for IgM, IgG and IgA antibodies against Cp were performed using the enzyme immunoassay (EIA) method. Univariate and multivariate analyses were carried out with respect to IgA, IgG, gender and type of cardiomyopathy. The IgA-IgG joint effect was also studied. The one-year survival rate was reported for patients with IgA < 8 EIU to be 72.2%, whereas those with IgA >or= 8 EIU accounted for only 43.5% (Kaplan-Meier analysis, p = 0.0548). In multivariate analysis IgA /IgG status proved to be a highly significant factor in survival. IgA positive outcome combined with IgG negative outcome showed that the relative risk of death equaled 12.08 versus other combinations of IgA/IgG status. In the Cox multivariate model ischemic cardiomyopathy showed a relative risk of 2.79 (p=0.0594), although it was not significant in univariate Chronic Cp infection, as expressed through a high IgA level, seems to have adverse impact on the survival rate in one-year follow-up after OHT. IgA titers against Cp in heart transplant recipients should therefore be assessed, as the high values might be a predictive risk factor within the first post-operative year.


Estimation of relevant variables on high-dimensional biological patterns using iterated weighted kernel functions.
The analysis of complex proteomic and genomic profiles involves the identification of significant markers within a set of hundreds or even thousands of variables that represent a high-dimensional problem space. The occurrence of noise, redundancy or combinatorial interactions in the profile makes the selection of relevant variables harder. Here we propose a method to select variables based on estimated relevance to hidden patterns. Our method combines a weighted-kernel discriminant with an iterative stochastic probability estimation algorithm to discover the relevance distribution over the set of variables. We verified the ability of our method to select predefined relevant variables in synthetic proteome-like data and then assessed its performance on biological high-dimensional problems. Experiments were run on serum proteomic datasets of infectious diseases. The resulting variable subsets achieved classification accuracies of 99% on Human African Trypanosomiasis, 91% on Tuberculosis, and 91% on Malaria serum proteomic profiles with fewer than 20% of variables selected. Our method scaled-up to dimensionalities of much higher orders of magnitude as shown with gene expression microarray datasets in which we obtained classification accuracies close to 90% with fewer than 1% of the total number of variables. Our method consistently found relevant variables attaining high classification accuracies across synthetic and biological datasets. Notably, it yielded very compact subsets compared to the original number of variables, which should simplify downstream biological experimentation.


Rapid increases in bat activity and diversity after wetland construction in an urban ecosystem.
Wetland construction can mitigate the biodiversity and water quality losses associated with reduced natural wetland coverage. While beneficial effects of wetland construction for bats have been observed in natural and rural settings, the effects of wetland construction on bats in an urban ecosystem are less understood. We used passive acoustic monitoring to measure bat activity levels and diversity at two constructed wetlands and two control sites on the University of North Carolina Greensboro campus, in Greensboro, North Carolina, USA. We monitored all 4 sites before and after wetland construction. Pre-wetland construction, there were few differences in bat activity and community structure at our sites. After wetland construction, we observed greater activity, attributable to all species we recorded, at wetland sites compared to control sites. Species diversity and species richness were also higher at wetland sites compared to control sites. When comparing the same sites before and after wetland construction, both bat activity and species richness increased after construction, but the effects were seen in Winter and not Spring. Our results demonstrate that bats use constructed wetlands in urban ecosystems similarly to other habitat settings. Increases in bat activity, diversity, and species richness occurred within one year of wetland construction.


High throughput LSPR and SERS analysis of aminoglycoside antibiotics.
Aminoglycoside antibiotics are used in the treatment of infections caused by Gram-negative bacteria, and are often dispensed only in severe cases due to their adverse side effects. Patients undergoing treatment with these antibiotics are therefore commonly subjected to therapeutic drug monitoring (TDM) to ensure a safe and effective personalised dosage. The ability to detect these antibiotics in a rapid and sensitive manner in human fluids is therefore of the utmost importance in order to provide effective monitoring of these drugs, which could potentially allow for a more widespread use of this class of antibiotics. Herein, we report on the detection of various aminoglycosides, by exploiting their ability to aggregate gold nanoparticles. The number and position of the amino groups of aminoglycoside antibiotics controlled the aggregation process. We investigated the complementary techniques of surface enhanced Raman spectroscopy (SERS) and localized surface plasmon resonance (LSPR) for dual detection of these aminoglycoside antibiotics and performed an in-depth study of the feasibility of carrying out TDM of tobramycin using a platform amenable to high throughput analysis. Herein, we also demonstrate dual detection of tobramycin using both LSPR and SERS in a single platform and within the clinically relevant concentration range needed for TDM of this particular aminoglycoside. Additionally we provide evidence that tobramycin can be detected in spiked human serum using only functionalised nanoparticles and SERS analysis.


Nonparametric method for genomics-based prediction of performance of quantitative traits involving epistasis in plant breeding.
Genomic selection (GS) procedures have proven useful in estimating breeding value and predicting phenotype with genome-wide molecular marker information. However, issues of high dimensionality, multicollinearity, and the inability to deal effectively with epistasis can jeopardize accuracy and predictive ability. We, therefore, propose a new nonparametric method, pRKHS, which combines the features of supervised principal component analysis (SPCA) and reproducing kernel Hilbert spaces (RKHS) regression, with versions for traits with no/low epistasis, pRKHS-NE, to high epistasis, pRKHS-E. Instead of assigning a specific relationship to represent the underlying epistasis, the method maps genotype to phenotype in a nonparametric way, thus requiring fewer genetic assumptions. SPCA decreases the number of markers needed for prediction by filtering out low-signal markers with the optimal marker set determined by cross-validation. Principal components are computed from reduced marker matrix (called supervised principal components, SPC) and included in the smoothing spline ANOVA model as independent variables to fit the data. The new method was evaluated in comparison with current popular methods for practicing GS, specifically RR-BLUP, BayesA, BayesB, as well as a newer method by Crossa et al., RKHS-M, using both simulated and real data. Results demonstrate that pRKHS generally delivers greater predictive ability, particularly when epistasis impacts trait expression. Beyond prediction, the new method also facilitates inferences about the extent to which epistasis influences trait expression.


Reconfigurable optical null based on counter-rotating Zernike plates for test of aspheres.
In off-axis subapertures of most aspheres, astigmatism and coma dominate the aberrations with approximately quadratic and linear increase as the off-axis distance increases. A pair of counter-rotating Zernike plates is proposed to generate variable amount of Zernike terms Z4 and Z6, correcting most of the astigmatism and coma for subapertures located at different positions on surfaces of various aspheric shapes. The residual subaperture aberrations are then reduced within the vertical dynamic range of measurement of the interferometer. The plates are fabricated with computer generated holograms and the experimental results show the variable aberration correction effect without ghost fringes. The same plates are reconfigurable by counter-rotating to enable near-null test of various aspheres flexibly.


Birth cohort patterns of gastric cancer and peptic ulcer among non-whites in the USA.
The aim of this study was to follow the long-term trends of mortality from gastric cancer, gastric ulcer and duodenal ulcer in the USA. It was hypothesised that the time trends among different ethnic groups would be shaped by similar birth cohort patterns. The US ulcer and cancer data were available from 1923 to 2006 and 1930 to 2006, respectively. Age-specific death rates were calculated for consecutive 10-year periods and 10-year age groups, stratified by sex and race. They were plotted against the period of death as period-age contours and against the period of birth as cohort-age contours. The time trends were also summarised as standardised cohort mortality ratios. The occurrence of birth cohort patterns with an initial rise and a subsequent decline was observed in gastric cancer and both ulcer types. It occurred similarly in white and non-white population, as well as men and women. In all ethnic groups alike, the initial rise in mortality from gastric cancer and gastric ulcer preceded the rise of duodenal ulcer by 10-20 years. The two time lags between gastric cancer and gastric ulcer and between gastric and duodenal ulcer were most pronounced in white patients. In non-white patients, the rise of gastric cancer occurred later than in white patients and coincided with that of gastric ulcer. The initial rise in mortality from gastric cancer was more evident in non-white than in white patients. The birth cohort pattern has shaped the time trends of Helicobacter pylori-related disease similarly among different ethnic and socioeconomic groups. Its cause remains unknown and warrants further investigation.


Advances in the management of MS symptoms: real-life evidence.
Once the efficacy and tolerability of a new intervention have been demonstrated in clinical trials under ideal experimental conditions, observational studies within approved label conditions are performed to determine its effectiveness in real-world use and expand the safety/tolerability data. Several large observational studies have been conducted of Sativex(®) (THC:CBD) oromucosal spray as add-on therapy for moderate-to-severe treatment-resistant multiple sclerosis (MS) spasticity. Studies performed to date with a focus on tolerability have confirmed the absence of abuse, misuse, addiction and lack of impairment of cognition or driving ability. Open-label studies performed to date with a focus on effectiveness have indicated that about one-half to two-thirds of patients initiated on THC:CBD oromucosal spray continue to derive benefit after 3 months' treatment at a mean dosage of 5-7 sprays/day.


Microstructured optical fiber coated with thin films for gas and chemical sensing.
We propose the use of tapered microstructured fibers with collapsed air-holes coated with thin layers for gas sensing. The collapsing of the holes allows having access to the evanescent fields which can be absorbed or attenuated with gas-permeable thin films. On the other hand, a section of the holey fiber is transformed into a solid multimode fiber. The beating between the multiple modes of the latter makes the transmission spectra of the device to exhibit an oscillatory pattern. This evanescent-fields-plus-modal-interferometer structure may offer interesting properties for gas and chemical sensing. As an example we demonstrate a hydrogen sensor.


Skin microdialysis: methods, applications and future opportunities-an EAACI position paper.
Skin microdialysis (SMD) is a versatile sampling technique that can be used to recover soluble endogenous and exogenous molecules from the extracellular compartment of human skin. Due to its minimally invasive character, SMD can be applied in both clinical and preclinical settings. Despite being available since the 1990s, the technique has still not reached its full potential use as a tool to explore pathophysiological mechanisms of allergic and inflammatory reactions in the skin. Therefore, an EAACI Task Force on SMD was formed to disseminate knowledge about the technique and its many applications. This position paper from the task force provides an overview of the current use of SMD in the investigation of the pathogenesis of chronic inflammatory skin diseases, such as atopic dermatitis, chronic urticaria, psoriasis, and in studies of cutaneous events during type 1 hypersensitivity reactions. Furthermore, this paper covers drug hypersensitivity, UVB-induced- and neurogenic inflammation, and drug penetration investigated by SMD. The aim of this paper is to encourage the use of SMD and to make the technique easily accessible by providing an overview of methodology and applications, supported by standardized operating procedures for SMD in vivo and ex vivo.


Wealth distribution under the spread of infectious diseases.
We develop a mathematical framework to study the economic impact of infectious diseases by integrating epidemiological dynamics with a kinetic model of wealth exchange. The multiagent description leads to the study of the evolution over time of a system of kinetic equations for the wealth densities of susceptible, infectious, and recovered individuals, whose proportions are driven by a classical compartmental model in epidemiology. Explicit calculations show that the spread of the disease seriously affects the distribution of wealth, which, unlike the situation in the absence of epidemics, can converge toward a stationary state with a bimodal form. Furthermore, simulations confirm the ability of the model to describe different phenomenon characteristics of economic trends in situations compromised by the rapid spread of an epidemic, such as the unequal impact on the various wealth classes and the risk of a shrinking middle class.


Structure of mitochondrial aldehyde dehydrogenase: the genetic component of ethanol aversion.
The single genetic factor most strongly correlated with reduced alcohol consumption and incidence of alcoholism is a naturally occurring variant of mitochondrial aldehyde dehydrogenase (ALDH2). This variant contains a glutamate to lysine substitution at position 487 (E487K). The E487K variant of ALDH2 is found in approximately 50% of the Asian population, and is associated with a phenotypic loss of ALDH2 activity in both heterozygotes and homozygotes. ALDH2-deficient individuals exhibit an averse response to ethanol consumption, which is probably caused by elevated levels of blood acetaldehyde. The structure of ALDH2 is important for the elucidation of its catalytic mechanism, to gain a clear understanding of the contribution of ALDH2 to the genetic component of alcoholism and for the development of specific ALDH2 inhibitors as potential drugs for use in the treatment of alcoholism. The X-ray structure of bovine ALDH2 has been solved to 2.65 A in its free form and to 2.75 A in a complex with NAD+. The enzyme structure contains three domains; two dinucleotide-binding domains and a small three-stranded beta-sheet domain, which is involved in subunit interactions in this tetrameric enzyme. The E487K mutation occurs in this small oligomerization domain and is located at a key interface between subunits immediately below the active site of another monomer. The active site of ALDH2 is divided into two halves by the nicotinamide ring of NAD+. Adjacent to the A-side (Pro-R) of the nicotinamide ring is a cluster of three cysteines (Cys301, Cys302 and Cys303) and adjacent to the B-side (Pro-S) are Thr244, Glu268, Glu476 and an ordered water molecule bound to Thr244 and Glu476. Although there is a recognizable Rossmann-type fold, the coenzyme-binding region of ALDH2 binds NAD+ in a manner not seen in other NAD+-binding enzymes. The positions of the residues near the nicotinamide ring of NAD+ suggest a chemical mechanism whereby Glu268 functions as a general base through a bound water molecule. The sidechain amide nitrogen of Asn169 and the peptide nitrogen of Cys302 are in position to stabilize the oxyanion present in the tetrahedral transition state prior to hydride transfer. The functional importance of residue Glu487 now appears to be due to indirect interactions of this residue with the substrate-binding site via Arg264 and Arg475.


Burnout and psychiatric disorder among cancer clinicians.
The prevalence and causes of 'burnout' and psychiatric disorder among senior oncologists and palliative care specialists have been measured in a national questionnaire-based survey. All consultant non-surgical oncologists in the UK were asked to participate. Sources of work-related stress and satisfaction were measured using study-specific questions which were aggregated into factors. Psychiatric disorder was estimated using the 12-item General Health Questionnaire. The three components of 'burnout'--emotional exhaustion, depersonalisation and low personal accomplishment--were assessed using the Maslach Burnout Inventory. Three hundred and ninety-three out of 476 (83%) consultants returned their questionnaires. The estimated prevalence of psychiatric disorder in cancer clinicians was 28%, and this is similar to the rate among British junior house officers. The study group had equivalent levels of emotional exhaustion and low personal accomplishment to those found in American doctors and nurses, but lower levels of depersonalisation. Among cancer clinicians, 'burnout' was more prevalent among clinical oncologists than among medical oncologists and palliative care specialists. Psychiatric disorder was independently associated with the stress of feeling overloaded (P < 0.0001), dealing with treatment toxicity/errors (P < 0.004) and deriving little satisfaction from professional status/esteem (P = 0.002). 'Burnout' was also related to these factors, and in addition was associated with high stress and low satisfaction from dealing with patients, and with low satisfaction from having adequate resources (each at a level of P < or = 0.002). Clinicians who felt insufficiently trained in communication and management skills had significantly higher levels of distress than those who felt sufficiently trained. If 'burnout' and psychiatric disorder among cancer clinicians are to be reduced, increased resources will be required to lessen overload and to improve training in communication and management skills.


Adipose tissue derived bacteria are associated with inflammation in obesity and type 2 diabetes.
Bacterial translocation to various organs including human adipose tissue (AT) due to increased intestinal permeability remains poorly understood. We hypothesised that: (1) bacterial presence is highly tissue specific and (2) related in composition and quantity to immune inflammatory and metabolic burden. We quantified and sequenced the bacterial 16S rRNA gene in blood and AT samples (omental, mesenteric and subcutaneous) of 75 subjects with obesity with or without type 2 diabetes (T2D) and used catalysed reporter deposition (CARD) - fluorescence in situ hybridisation (FISH) to detect bacteria in AT. Under stringent experimental and bioinformatic control for contaminants, bacterial DNA was detected in blood and omental, subcutaneous and mesenteric AT samples in the range of 0.1 to 5 pg/µg DNA isolate. Moreover, CARD-FISH allowed the detection of living, AT-borne bacteria. Proteobacteria and Firmicutes were the predominant phyla, and bacterial quantity was associated with immune cell infiltration, inflammatory and metabolic parameters in a tissue-specific manner. Bacterial composition differed between subjects with and without T2D and was associated with related clinical measures, including systemic and tissues-specific inflammatory markers. Finally, treatment of adipocytes with bacterial DNA in vitro stimulated the expression of TNFA and IL6. Our study provides contaminant aware evidence for the presence of bacteria and bacterial DNA in several ATs in obesity and T2D and suggests an important role of bacteria in initiating and sustaining local AT subclinical inflammation and therefore impacting metabolic sequelae of obesity.


High-resolution transcriptome and genome-wide dynamics of RNA polymerase and NusA in Mycobacterium tuberculosis.
To construct a regulatory map of the genome of the human pathogen, Mycobacterium tuberculosis, we applied two complementary high-resolution approaches: strand-specific RNA-seq, to survey the global transcriptome, and ChIP-seq, to monitor the genome-wide dynamics of RNA polymerase (RNAP) and the anti-terminator NusA. Although NusA does not bind directly to DNA, but rather to RNAP and/or to the nascent transcript, we demonstrate that NusA interacts with RNAP ubiquitously throughout the chromosome, and that its profile mirrors RNAP distribution in both the exponential and stationary phases of growth. Generally, promoter-proximal peaks for RNAP and NusA were observed, followed by a decrease in signal strength reflecting transcriptional polarity. Differential binding of RNAP and NusA in the two growth conditions correlated with transcriptional activity as reflected by RNA abundance. Indeed, a significant association between expression levels and the presence of NusA throughout the gene body was detected, confirming the peculiar transcription-promoting role of NusA. Integration of the data sets pinpointed transcriptional units, mapped promoters and uncovered new anti-sense and non-coding transcripts. Highly expressed transcriptional units were situated mainly on the leading strand, despite the relatively unbiased distribution of genes throughout the genome, thus helping the replicative and transcriptional complexes to align.


The Best of Both Worlds: The Role of Career Adaptability and Career Competencies in Students' Well-Being and Performance.
In addition to acquiring occupation-specific knowledge and skills, students need to develop a set of career self-management skills - or resources - that helps them to successfully maneuver the various career-related challenges they face and that stimulate their well-being, engagement, and performance in studying tasks. In the current study, we apply the Job Demands-Resources (JD-R) theory in an educational setting and suggest that career adaptability and career competencies are important career resources that predict both life satisfaction and academic performance via students' satisfaction with the choice of their major and study engagement. Undergraduate students (N = 672) from nine different colleges and universities in Lithuania participated in the study. The results revealed that career adaptability and career competencies were positively linked to students' life satisfaction, both directly and via study engagement. In addition, these career resources were positively, yet indirectly, related to academic performance via study engagement. Overall, the results suggest that career resources contribute to study engagement, life satisfaction, and academic performance. The results of our study further support JD-R theorizing and its applicability in student samples. Further theoretical and practical implications are discussed.


A family of cis-macrocyclic diphosphines: modular, stereoselective synthesis and application in catalytic CO2/ethylene coupling.
A family of cis-macrocyclic diphosphines was prepared in just three steps from white phosphorus and commercial materials using a modular synthetic approach. Alkylation of bicyclic diphosphane 3,4,8,9-tetramethyl-1,6-diphosphabicyclo(4.4.0)deca-3,8-diene, or P2(dmb)2, produced phosphino-phosphonium salts [R-P2(dmb)2]X, where R is methyl, benzyl and isobutyl, in yields of 90-96%. Treatment of these salts with organolithium or Grignard reagents yielded symmetric and unsymmetric macrocyclic diphosphines of the form cis-1-R-6-R'-3,4,8,9-tetramethyl-2,5,7,10-tetrahydro-1,6-DiPhospheCine, or R,R'-DPC, in which R' is methyl, cyclohexyl, phenyl or mesityl, in yields of 46-94%. Alternatively, symmetric diphosphine Cy2-DPC was synthesized in 74% yield from the dichlorodiphosphine Cl2P2(dmb)2. As a first application, these cis-macrocyclic diphosphines were used as ligands in the nickel-catalyzed synthesis of acrylate from CO2 and ethylene, for which they showed promising catalytic activity.


Non-target effects of clothianidin on monarch butterflies.
Monarch butterflies (Danaus plexippus) frequently consume milkweed in and near agroecosystems and consequently may be exposed to pesticides like neonicotinoids. We conducted a dose response study to determine lethal and sublethal doses of clothianidin using a 36-h exposure scenario. We then quantified clothianidin levels found in milkweed leaves adjacent to maize fields. Toxicity assays revealed LC10, LC50, and LC90 values of 7.72, 15.63, and 30.70 ppb, respectively. Sublethal effects (larval size) were observed at 1 ppb. Contaminated milkweed plants had an average of 1.14±0.10 ppb clothianidin, with a maximum of 4 ppb in a single plant. This research suggests that clothianidin could function as a stressor to monarch populations.


COVID-19 vaccination rates, intent, and hesitancy in patients with solid organ and blood cancers: A multicenter study.
People with cancer are at higher risk of serious illness and death from COVID-19 infection. We investigated COVID-19 vaccine uptake among patients with solid organ and blood cancers and explored factors related to hesitancy. Cross-sectional online survey of adults with a history of cancer at three health services across metropolitan and regional Victoria. Vaccine hesitancy was measured by the validated Oxford COVID-19 vaccine hesitancy scale. There were 1073 respondents: 56% female; median age 62 years (range 23 - 91). Commonest tumor types included breast 29%, gastrointestinal 19%, hematological 15%, genitourinary 15%, and lung 8%. Thirty-six percent had metastatic disease, and 54% were receiving active anticancer treatment. Eighty-four percent of respondents indicated positive intent toward COVID-19 vaccination, 10% were undecided, and 6% indicated negative attitudes. At least one vaccine dose had been received by 65% of respondents, leaving 35% unvaccinated. Fifty-eight percent of unvaccinated patients answered that they would "definitely" or "probably" take a vaccine. Higher vaccine uptake was significantly associated with older age, male gender, English as first language, longer time since cancer diagnosis, and not being on current anticancer treatment. Concerns regarding vaccine side effects, particularly thrombosis, and the desire for clear medical advice were prominent among unvaccinated respondents. Despite being eligible for COVID-19 vaccination since March 2021, a substantial minority of patients with cancer remained unvaccinated as of August 2021. Targeted communication and educational resources addressing vaccine safety in the context of cancer are key to promoting vaccine uptake in this vulnerable population.


The effects of walking and cycling computer workstations on keyboard and mouse performance.
This study aimed to determine the effects of active workstation designs on speed and error during typing, mouse pointing, and combined type and mouse-use tasks. Office ergonomics has focused on musculoskeletal disorder prevention; however, increasing computer-based work also increases health risks associated with inactivity. Workstations allowing computer users to walk or cycle while performing computer tasks have been shown to demand sufficient energy expenditure to result in significant health benefits. However the performance effects of being active while using a computer have not been documented. Thirty office workers (16 female, 15 touch typists) performed standardized computer tasks in six workstation conditions: sitting, standing, walking at 1.6 km/h and 3.2 km/h, and cycling at 5 and 30 watts. Performance, perceived performance, and heart rate were measured. Computer task performance was lower when walking and slightly lower when cycling, compared with chair sitting. Standing performance was not different from sitting performance. Mouse performance was more affected than typing performance. Performance decrements were equal for females and males and for touch typists and nontouch typists. Performance decrements maybe related to both biomechanical and cognitive processes. Active workstations may be less suitable for mouse-intensive work and susceptible users. Although active workstations may result in some decrement in performance, their ability to increase daily energy expenditure may make them a feasible solution for workplace inactivity.


Safety and efficacy of autologous hematopoietic stem-cell transplantation following natalizumab discontinuation in aggressive multiple sclerosis.
Natalizumab (NTZ) is a highly effective treatment for relapsing-remitting multiple sclerosis (MS), but its withdrawal is often followed by disease reactivation or rebound, even if other disease-modifying treatments (DMTs) are administered. In this study, for the first time, the safety and efficacy of autologous hematopoietic stem-cell transplantation (aHSCT) performed following NTZ discontinuation were retrospectively compared with conventional DMTs. Patients with relapsing-remitting MS treated with NTZ who discontinued the drug after at least six administrations and with at least 6 months of follow-up were included. Patients underwent aHSCT after a minimum of 6 months following NTZ withdrawal, receiving meanwhile cyclophosphamide or corticosteroids, or other DMTs approved for MS (control group) after an adequate wash-out period. Both hematological and neurological follow-up were assessed according to standard policies. A total of 52 patients were included, 11 who received aHSCT and 41 who received DMTs. Baseline clinical and demographic characteristics were similar between the two groups. No fatality or life-threatening complications, including progressive multifocal leukoencephalopathy, were observed. At 3 years following NTZ discontinuation, no evidence of disease activity was reported in 54.5% of the patients in the aHSCT group compared with 11.5% of those in the DMT group (P = 0.0212). Disease reactivation in the patients with aHSCT was observed only during wash-out/bridging therapy and 100% of the cases were free from disease activity after aHSCT. These data suggest that an aggressive therapy should be established after NTZ with the shortest possible wash-out period. aHSCT after 6 months from NTZ withdrawal appears to be safe.


Adaptation and validation of a bacteria-specific enzyme-linked immunosorbent assay for determination of farm-specific Lawsonia intracellularis seroprevalence in central Kentucky Thoroughbreds.
Lawsonia intracellularis is the causative agent of equine proliferative enteropathy (EPE), a disease for which no large-scale seroprevalence studies have been conducted. To validate and use an equine-specific enzyme-linked immunosorbent assay (ELISA) for L. intracellularis to determine the seroprevalence of L. intracellularis on numerous farms. An ELISA, in which purified antigen was used, was adapted from previous work in swine. A total of 337 Thoroughbreds from 25 central Kentucky farms were enrolled and monthly serum samples collected from August 2010 to January/February 2011. Samples were screened for L. intracellularis-specific antibodies using a modified ELISA. Farms were classified into one of 3 groups based on 3 year prior history with EPE. The ELISA intra-assay coefficient of variation (CV) was 6.73 and inter-assay CV was 9.60. An overall seroprevalence of 68% was obtained, with farm-specific seroprevalances ranging from 14 to 100%. A significant difference was found in the average seroprevalence (P<0.05) on farms with a confirmed recent history of EPE cases. Additionally, both lower average ELISA unit (EU) values (P = 0.079) and maximum EU values (P = 0.056) were detected on farms with no recent EPE history when compared to the other groups. A bimodal exposure distribution to L. intracellularis was detected in the fall and winter months. Recent history of EPE was associated with higher average seroprevalence indicating increased exposure on farms with prior cases of EPE. Seasonally bimodal exposure was also observed. The adapted ELISA appears to be useful for determination of L. intracellularis-specific antibody levels. The high farm-specific seroprevalences and bimodal distribution of exposure to L. intracellularis were unexpected and suggest that farms with a previous history of EPE remain at risk due to heightened exposure levels beyond early winter.


Neurasthenia and depression: a study of somatization and culture in China.
The author reviews conceptual and empirical issues regarding the interaction of neurasthenia, somatization and depression in Chinese culture and in the West. The historical background of neurasthenia and its current status are discussed, along with the epidemiology and phenomenology of somatization and depression. Findings are presented from a combined clinical and anthropological field study of 100 patients with neurasthenia in the Psychiatry Outpatient Clinic at the Hunan Medical College. Eighty-seven of these patients made the DSM-III criteria of Major Depressive Disorder; diagnoses of anxiety disorders were also frequent. Forty-four patients were suffering from chronic pain syndromes previously undiagnosed, and cases of culture-bound syndromes also were detected. For three-quarters of patients the social significances and uses of their illness behavior chiefly related to work. Although from the researcher's perspective 70% of patients with Major Depressive Disorder experienced substantial improvement and 87% some improvement in symptoms when treated with antidepressant medication, fewer experienced decreased help seeking, and a much smaller number perceived less social impairment and improvement in illness problems (the psychosocial accompaniment of disease including maladaptive coping and work, family and school problems). These findings are drawn on to advance medical anthropology and cultural psychiatry theory and research regarding somatization in Chinese culture, the United States and cross culturally. The author concludes that though neurasthenia can be understood in several distinctive ways, it is most clinically useful to regard it as bioculturally patterned illness experience (a special form of somatization) related to either depression and other diseases or to culturally sanctioned idioms of distress and psychosocial coping.


Comparative digestion studies in wild suids at Rotterdam Zoo.
Among the artiodactyla, the suids are a group whose digestive physiology has hardly been investigated. The apparent digestibilities (aD) of macronutrients were measured in captive specimens of warthog (Phacochoerus africanus), red river hog (Potamochoerus porcus), and Visayan warty pigs (Sus cebifrons), and compared with those reported for babirusa (Babyrousa babyrussa) from the same facility on a similar diet. The animals were fed mixed diets of pelleted feed, grains, fruits, and vegetables; dietary neutral detergent fiber (NDF) ranged from 17 to 26% dry matter. aD of organic matter and protein ranged from 72 to 89 and 70 to 82%, respectively. Although red river hogs and warty pigs achieved aD of NDF of 41-54%, this value was higher both in warthogs (63-66%) and babirusa (61-63%). aD of acid detergent fiber was comparatively low in red river hogs (4%), warty pigs (22%), and babirusa (13-25%) but high in warthog (59-62%). Comparison with additional literature data (including peccaries) indicates that in spite of differences in digestive anatomy, suids and peccaries are similar, and resemble other herbivores in fundamental characteristics, such as the negative influence of fiber on overall digestibility, or the positive influence of dietary protein on protein digestion. Although the existing data are equivocal as to a superior fiber digestion in peccaries as compared with other wild suids, the results suggest that warthogs are more efficient than other wild suids or peccaries in terms of fiber digestion. Zoo Biol 27:305-319, 2008. (c) 2008 Wiley-Liss, Inc.


Stimulation of MAP kinase by v-raf transformation of fibroblasts fails to induce hyperphosphorylation of transfected tau.
A proportion of the microtubule-associated protein, tau, is in an elevated state of phosphorylation in foetal and adult brain whereas all of the tau in paired helical filaments, which are characteristic of Alzheimer's disease is hyperphosphorylated; it is important therefore to elucidate the mechanisms that regulate tau phosphorylation. Here we describe results that show that although MAP kinase can hyperphosphorylate tau in vitro, activation of MAP kinase in transformed fibroblasts does not result in hyperphosphorylation of transfected tau, whereas glycogen synthase kinase-3 beta (GSK-3 beta) when co-transfected with tau does result in tau hyperphosphorylation. The findings imply that GSK-3 beta may be a stronger candidate than MAP kinase for inducing tau hyperphosphorylation in vivo.


Antibodies bound to lipid haptens in model membranes diffuse as rapidly as the lipids themselves.
A pattern photobleaching method has been used to measure the rates of lateral diffusion of fluorescent-labeled specific anti-nitroxide IgG bound with both combining sites to nitroxide-containing phospholipids in liposomal membranes composed of dimyristoyl phosphatidylcholine at 28 degrees C ("fluid"), dipalmitoyl phosphatidylcholine at 32 degrees C ("Solid"), and dipalmitoyl phosphatidylcholine containing 15 or 25 mol% cholesterol ("solid" or "fluid," respectively, at 32 degrees C). The diffusion coefficients of the bound immunoglobulin were found to be the same as those of fluorescent-labeled phospholipids in each case even though these diffusion coefficients range from 10(-11) to 10(-8) cm2/sec. Hapten-containing liposomal membranes of the type studied here have previously been shown to elicit a number of antibody-dependent immune responses. Therefore, this work indicates that membrane-bound but otherwise freely diffusing antibodies are sufficient for these reponses.


Novel 3D collagen scaffolds fabricated by indirect printing technique for tissue engineering.
This article reports the mechanical properties and in vitro evaluation of a collagen scaffold fabricated using an indirect 3D printing technique. Collagen scaffolds, featuring predefined internal channels and capillary networks, were manufactured using phase change printing. It was observed that the collagen scaffolds featured internal channels and a hierarchical structure that varied over length scales of 10-400 microm. In vitro evaluation using hMSCs demonstrated that the resultant collagen based scaffolds have the ability to support hMSC cell attachment and proliferation; cells can migrate and survive deep within the structure of the scaffold. The cell numbers increased 2.4 times over 28 days in culture for the lysine treated scaffolds. The cells were spread along the collagen fibers to form a 3D structure and extracellular matrix was detected on the surface of the scaffolds after 4 weeks in culture. The crosslinking treatment enhanced the biostability and dynamic properties of the collagen scaffolds significantly.


Effects of a Change from an Indoor-Based Total Mixed Ration to a Rotational Pasture System Combined with a Moderate Concentrate Feed Supply on Immunological Cell and Blood Parameters of Dairy Cows.
In spring, transition from a total mixed ration (TMR) to a full grazing ration with moderate concentrate supply influences cow's metabolism. It has been shown that feeding moderate amounts of concentrate during fulltime grazing did not prevent energy shortage and lipomobilization, alterations in energy metabolism, decreasing milk production and loss in body weight. As diet change and energy balance are closely related to immune reactivity, in this trial the effect of transition to pasture on specific immune parameters of cows was documented. Over a 12-week trial 43 dairy cows were observed during transition from confinement to pasture (PG; n = 22) and compared to cows fed TMR indoor (CG; n = 21). The CG stayed on a TMR based ration (35% corn silage, 35% grass silage, 30% concentrate; dry matter (DM) basis), whereas the PG slowly switched to a pasture -based ration (week 0 and 1 = TMR, week 2 = TMR and 3 h pasture·day-1, week 3 and 4 = TMR and 12 h pasture·day-1, and week 5 to 11 = pasture combined with 4.5 kg DM concentrate·cow-1·day-1). Inflammatory markers like blood haptoglobin or tryptophan to kynurenine ratio did not indicate acute phase reaction. Proportions of CD4+ (T-helper cells) and CD8+ cells (cytotoxic T-cells) remained uninfluenced as well. White blood cell concentration and its subpopulation of granulocytes increased over time in the PG. Stimulation ability of peripheral blood mononuclear cells to mount an oxidative burst significantly increased during the trial, too. The endogenous antioxidant state as characterized by glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity in blood of the PG did not change, whereas the vitamin E concentration reached the highest level at the end of the trial. The 25-CHO metabolites of vitamin D increased as soon as the PG had pasture access, whereas the other metabolite 25-ERG decreased. The results of this study indicate that transition to pasture affects immune related parameters. However, the consequences of the observed effects on health status of the pasture group need to be clarified in further studies with a defined concurrent immune challenge.


Tat expression led to increased histone 3 tri-methylation at lysine 27 and contributed to HIV latency in astrocytes through regulation of MeCP2 and Ezh2 expression.
Astrocytes are susceptible to HIV infection and potential latent HIV reservoirs. Tat is one of three abundantly expressed HIV early genes in HIV-infected astrocytes and has been shown to be a major pathogenic factor for HIV/neuroAIDS. In this study, we sought to determine if and how Tat expression would affect HIV infection and latency in astrocytes. Using the glycoprotein from vesicular stomatitis virus-pseudotyped red-green HIV (RGH) reporter viruses, we showed that HIV infection was capable of establishing HIV latency in astrocytes. We also found that Tat expression decreased the generation of latent HIV-infected cells. Activation of latent HIV-infected astrocytes showed that treatment of GSK126, a selective inhibitor of methyltransferase enhancer of zeste homolog 2 (Ezh2) that is specifically responsible for tri-methylation of histone 3 lysine 27 (H3K27me3), led to activation of significantly more latent HIV-infected Tat-expressing astrocytes. Molecular analysis showed that H3K27me3, Ezh2, MeCP2, and Tat all exhibited a similar bimodal expression kinetics in the course of HIV infection and latency in astrocytes, although H3K27me3, Ezh2, and MeCP2 were expressed higher in Tat-expressing astrocytes and their expression were peaked immediately preceding Tat expression. Subsequent studies showed that Tat expression alone was sufficient to induce H3K27me3 expression, likely through its regulation of Ezh2 and MeCP2 expression. Taken together, these results showed for the first time that Tat expression induced H3K27me3 expression and contributed to HIV latency in astrocytes and suggest a new role and novel mechanism for Tat in HIV latency.


Handheld office-based spirometry versus laboratory spirometry in low-risk patients undergoing lung resection.
To compare handheld office spirometry with laboratory spirometry in evaluation of low-risk patients undergoing pulmonary resection. Low-risk patients evaluated for lung resection were enrolled in a prospective study and underwent office and laboratory spirometry. Standard laboratory spirometry values were considered the gold standard. Values of forced expiratory volume in 1 second (FEV1) <1.5 L or FEV1% <60% were determined in advance to signal high risk. Thirty patients with mean age 64 ± 12 years and median Karnofsky performance status 100 (80-100) were evaluated. The mean FEV1-office spirometry and mean FEV1-laboratory spirometry were comparable (2.14 L vs 2.36 L) as were the mean FEV1%-office spirometry and FEV1%-laboratory spirometry (78% vs 85%). The differences were within limits established a priori. Laboratory spirometry identified 3 of 30 (10%) patients as high risk and office spirometry identified 5 of 30 (16%) patients as high risk, including the 3 patients identified by laboratory spirometry. The sensitivity of office spirometry for detecting high risk was 100% (3/3) and specificity was 93% (25/27). The negative predictive value of office spirometry was 100% (25/25). Office spirometry added a median of 1 minute to the office visit, whereas laboratory spirometry added 42 minutes. Eventually, 25 of 30 (83%) patients underwent lung resection. There were 12 lobar and 13 sublobar resections. One patient developed postoperative respiratory morbidity. This patient was high risk by office spirometry but not by laboratory spirometry. This patient was discharged on home oxygen. Office spirometry FEV1 and FEV1% are clinically comparable to formal laboratory spirometry values. Office Spirometry saves time and possibly expense in the preoperative workup of patients evaluated for lung resection.


A model approach to project the start of egg laying of Great Tit (Parus major L.) in response to climate change.
The aim of this study was to select a phenological model that is able to calculate the beginning of egg laying of Great Tit (Parus major) for both current and future climate conditions. Four models (M1-M4) were optimised on long-term phenological observations from the Ecological Research Centre Schlüchtern (Hessen/Germany). Model M1 was a common thermal time model that accumulates growing degree days (GDD) on an optimised starting date t (1). Since egg laying of Great Tit is influenced not only by air temperature but also by photoperiod, model M1 was extended by a daylength term to give M2. The other two models, M3 and M4, correspond to M1 and M2, but t (1) was intentionally set to 1 January, in order to consider already rising temperatures at the beginning of the year. A comparison of the four models led to following results: model M1 had a relatively high root mean square error at verification (RMSE(ver)) of more than 4 days and can be used only to calculate the start of egg laying for current climate conditions because of the relatively late starting date for GDD calculation. The model failed completely if the starting date was set to 1 January (M3). Consideration of a daylength term in models M2 and M4 improved the performance of both models strongly (RMSE(ver) of only 3 days or less), increased the credibility of parameter estimation, and was a precondition to calculate reliable projections in the timing of egg laying in birds for the future. These results confirm that the start of egg laying of Great Tit is influenced not only by air temperature, but also by photoperiod. Although models M2 and M4 both provide comparably good results for current climate conditions, we recommend model M4-with a starting date of temperature accumulation on 1 January-for calculating possible future shifts in the commencement of egg laying. Our regional projections in the start of egg laying, based on five regional climate models (RCMs: REMO-UBA, ECHAM5-CLM, HadCM3-CLM, WETTREG-0, WETTREG-1, GHG emission scenario A1B), indicate that in the near future (2011-2040) no significant change will take place. However, in the mid- (2041-2070) and long-term (2071-2100) range the beginning of egg laying could be advanced significantly by up to 11 days on average of all five RCMs. This result corresponds to the already observed shift in the timing of egg laying by about 1 week, due mainly to an abrupt increase in air temperature at the end of the 1980s by 1.2 K between April and May. The use of five regional climate scenarios additionally allowed to estimate uncertainties among the RCMs.


Induction of NR4A orphan nuclear receptor expression in macrophages in response to inflammatory stimuli.
Oxidized lipids and inflammatory cytokines are believed to play a causal role in atherosclerosis through the regulation of gene expression in macrophages and other cells. Previous work has implicated the nuclear receptors peroxisome proliferator-activated receptor and liver X receptor in the control of lipid-dependent gene expression and inflammation. Here we demonstrate that expression of a third group of nuclear receptors, the NR4A ligand-independent orphan receptors, is highly inducible in macrophages by diverse inflammatory stimuli. Treatment of macrophages with lipopolysaccharide (LPS), cytokines, or oxidized lipids triggers the transcriptional induction of Nur77 (NR4A1), Nurr1 (NR4A2), and NOR1 (NR4A3) expression. Several lines of evidence point to the NF-kappaB signaling pathway as a principal mediator of inducible NR4A expression in macrophages. Analysis of the murine and human Nur77 promoters revealed two highly conserved NF-kappaB response elements. Mutation of these elements inhibited LPS-dependent expression of the Nur77 promoter in transient transfection assays. Furthermore, induction of Nur77 expression by LPS was severely compromised in fibroblasts lacking the three NF-kappaB subunits, Nfkb1, c-Rel, and RelA. Consistent with its ability to be induced by oxidized lipids, Nur77 was expressed in macrophages within human atherosclerotic lesions. These results identified NR4A nuclear receptors as potential transcriptional mediators of inflammatory signals in activated macrophages.