Incidence of paradoxical tuberculosis-associated immune reconstitution inflammatory syndrome and impact on patient outcome.
We used data from a randomized trial of HIV-tuberculosis co-infected patients in Mozambique to determine the incidence and predictors of paradoxical tuberculosis-associated immune reconstitution inflammatory syndrome (IRIS) occurring within 12 weeks of starting antiretroviral therapy, and to evaluate its association with patient outcome at 48 weeks. HIV-tuberculosis co-infected and antiretroviral therapy-naïve adults with less than 250 CD4/mm3 were randomized to a nevirapine or efavirenz-based antiretroviral therapy initiated 4 to 6 weeks after starting tuberculosis treatment, and were then followed for 48 weeks. Tuberculosis cases were diagnosed using WHO guidelines, and tuberculosis-IRIS by case definitions of the International Network for the Study of HIV-associated IRIS. The 573 HIV-tuberculosis co-infected patients who initiated antiretroviral therapy had a median CD4 count of 92 cells/mm(3) and HIV-1 RNA of 5.6 log10 copies/mL. Mortality at week 48 was 6.1% (35/573). Fifty-three (9.2%) patients presented a tuberculosis-IRIS within 12 weeks of starting antiretroviral therapy. Being female and having a low CD4 count, high HIV-1 RNA load, low body mass index and smear-positive pulmonary tuberculosis were independently associated with tuberculosis-IRIS. After adjustment for baseline body mass index, CD4 count and hemoglobin, occurrence of tuberculosis-IRIS was independently associated with 48-week mortality (aOR 2.72 95%CI 1.14-6.54). Immunological and HIV-1 virological responses and tuberculosis treatment outcomes were not different between patients with and without tuberculosis-IRIS. In this large prospective cohort, tuberculosis-IRIS occurrence within 12 weeks of starting antiretroviral therapy was independently associated with the mortality of HIV-tuberculosis co-infected patients at 48 weeks post antiretroviral therapy initiation.


A compact VEGF signature associated with distant metastases and poor outcomes.
Tumor metastases pose the greatest threat to a patient's survival, and thus, understanding the biology of disseminated cancer cells is critical for developing effective therapies. Microarrays and immunohistochemistry were used to analyze primary breast tumors, regional (lymph node) metastases, and distant metastases in order to identify biological features associated with distant metastases. When compared with each other, primary tumors and regional metastases showed statistically indistinguishable gene expression patterns. Supervised analyses comparing patients with distant metastases versus primary tumors or regional metastases showed that the distant metastases were distinct and distinguished by the lack of expression of fibroblast/mesenchymal genes, and by the high expression of a 13-gene profile (that is, the 'vascular endothelial growth factor (VEGF) profile') that included VEGF, ANGPTL4, ADM and the monocarboxylic acid transporter SLC16A3. At least 8 out of 13 of these genes contained HIF1alpha binding sites, many are known to be HIF1alpha-regulated, and expression of the VEGF profile correlated with HIF1alpha IHC positivity. The VEGF profile also showed prognostic significance on tests of sets of patients with breast and lung cancer and glioblastomas, and was an independent predictor of outcomes in primary breast cancers when tested in models that contained other prognostic gene expression profiles and clinical variables. These data identify a compact in vivo hypoxia signature that tends to be present in distant metastasis samples, and which portends a poor outcome in multiple tumor types.This signature suggests that the response to hypoxia includes the ability to promote new blood and lymphatic vessel formation, and that the dual targeting of multiple cell types and pathways will be needed to prevent metastatic spread.


The effects of sampling strategy on the quality of reconstruction of viral population dynamics using Bayesian skyline family coalescent methods: A simulation study.
The ongoing large-scale increase in the total amount of genetic data for viruses and other pathogens has led to a situation in which it is often not possible to include every available sequence in a phylogenetic analysis and expect the procedure to complete in reasonable computational time. This raises questions about how a set of sequences should be selected for analysis, particularly if the data are used to infer more than just the phylogenetic tree itself. The design of sampling strategies for molecular epidemiology has been a neglected field of research. This article describes a large-scale simulation exercise that was undertaken to select an appropriate strategy when using the GMRF skygrid, one of the Bayesian skyline family of coalescent methods, in order to reconstruct past population dynamics. The simulated scenarios were intended to represent sampling for the population of an endemic virus across multiple geographical locations. Large phylogenies were simulated under a coalescent or structured coalescent model and sequences simulated from these trees; the resulting datasets were then downsampled for analyses according to a variety of schemes. Variation in results between different replicates of the same scheme was not insignificant, and as a result, we recommend that where possible analyses are repeated with different datasets in order to establish that elements of a reconstruction are not simply the result of the particular set of samples selected. We show that an individual stochastic choice of sequences can introduce spurious behaviour in the median line of the skygrid plot and that even marginal likelihood estimation can suggest complicated dynamics that were not in fact present. We recommend that the median line should not be used to infer historical events on its own. Sampling sequences with uniform probability with respect to both time and spatial location (deme) never performed worse than sampling with probability proportional to the effective population size at that time and in that location and frequently was superior. As a result, we recommend this approach in the design of future studies. We also confirm that the inclusion of many recent sequences from a single geographical location in an analysis tends to result in a spurious bottleneck effect in the reconstruction and caution against interpreting this as genuine.


Amino acid digestibility and energy concentration in a novel source of high-protein distillers dried grains and their effects on growth performance of pigs.
Three experiments were conducted to evaluate a novel source of high-protein distillers dried grains produced by Buhler Inc. (HP-DDG(Buhler)) and fed to growing pigs. The standardized ileal digestibility (SID) of AA in HP-DDG(Buhler) and in soybean meal (SBM) was determined in Exp. 1. Nine pigs (109.8 +/- 2.78 kg of BW) were fitted with a T-cannula in the distal ileum and allotted to a triplicated 3 x 3 Latin square design with 3 diets and 3 periods per square. Diets containing HP-DDG(Buhler) or SBM as the only source of AA and an N-free diet were formulated. The SID of indispensable AA was less (P < 0.01) in HP-DDG(Buhler) than in SBM (Arg, 87.5 vs. 93.9%; His, 76.7 vs. 88.7%; Ile, 76.4 vs. 87.5%; Leu, 77.8 vs. 86.8%; Lys, 75.4 vs. 88.4%; Met, 82.8 vs. 88.4%; Phe, 77.9 vs. 87.3%; Thr, 72.5 vs. 83.5%; Trp, 85.1 vs. 91.0%; Val, 73.3 vs. 84.3%). The DE and ME in HP-DDG(Buhler) and in corn were measured in Exp. 2 using 16 growing barrows (24.6 +/- 1.66 kg of BW). A corn-based diet and a diet containing 50% corn and 48.2% HP-DDG(Buhler) were formulated. The total collection method and the difference procedures were used. The concentrations of DE and ME in HP-DDG(Buhler) were greater (P < 0.001) than in corn (5,043 vs. 4,002 kcal/kg of DM and 4,690 vs. 3,921 kcal/kg of DM, respectively). Experiment 3 was a 9-wk growth assay using 40 pigs (initial BW: 58.2 +/- 2.28 kg) allotted to 5 dietary treatments, with 8 replicates of individually housed pigs per treatment. Treatments included a control diet based on corn and SBM and 4 diets in which HP-DDG(Buhler) replaced 33, 66, 66, or 100% of the SBM in the control diet. All HP-DDG(Buhler) diets contained supplemental Lys and Thr to provide similar concentrations of SID Lys and Thr in all diets; one of the diets in which HP-DDG(Buhler) replaced 66% of the SBM and the diet in which HP-DDG(Buhler) replaced 100% of the SBM also contained crystalline Trp. Dietary treatments had no effect on ADG (1.15, 1.13, 1.16, 1.12, and 1.14 kg), ADFI (3.33, 3.35, 3.39, 3.30, and 3.33 kg), or G:F (0.35, 0.34, 0.34, 0.34, and 0.34 kg/kg). Carcass traits of pigs fed the diet in which HP-DDG(Buhler) replaced 100% of the SBM were not different from those of pigs fed the control diet. In conclusion, HP-DDG(Buhler) contains more DE and ME than corn, but has decreased SID values for AA compared with SBM. Soybean meal can be replaced by HP-DDG(Buhler) in diets fed to finishing pigs without any effect on growth performance or carcass characteristics, provided that diets are adequate in indispensable AA.


Alu repeats as transcriptional regulatory platforms in macrophage responses to M. tuberculosis infection.
To understand the epigenetic regulation of transcriptional response of macrophages during early-stage M. tuberculosis (Mtb) infection, we performed ChIPseq analysis of H3K4 monomethylation (H3K4me1), a marker of poised or active enhancers. De novo H3K4me1 peaks in infected cells were associated with genes implicated in host defenses and apoptosis. Our analysis revealed that 40% of de novo regions contained human/primate-specific Alu transposable elements, enriched in the AluJ and S subtypes. These contained several transcription factor binding sites, including those for members of the MEF2 and ATF families, and LXR and RAR nuclear receptors, all of which have been implicated in macrophage differentiation, survival, and responses to stress and infection. Combining bioinformatics, molecular genetics, and biochemical approaches, we linked genes adjacent to H3K4me1-associated Alu repeats to macrophage metabolic responses against Mtb infection. In particular, we show that LXRα signaling, which reduced Mtb viability 18-fold by altering cholesterol metabolism and enhancing macrophage apoptosis, can be initiated at response elements present in Alu repeats. These studies decipher the mechanism of early macrophage transcriptional responses to Mtb, highlighting the role of Alu element transposition in shaping human transcription programs during innate immunity.


Hormone therapy and cerebrovascular events: a population-based nested case-control study.
The relationship between postmenopausal hormone therapy (HT) and cerebrovascular disease has been examined in several epidemiological studies and clinical trials with conflicting results. The authors aimed to evaluate the association between the use of HT and the incidence of first cerebrovascular event. The study cohort comprised 158,031 women 50 to 69 years old registered in the U.K. General Practice Research Database between 1991 and 1997. The authors conducted a nested case-control analysis using all 920 confirmed cases of cerebrovascular events identified during the follow-up (536 of transient ischemic attack [TIA]; 259 of ischemic stroke; 125 of hemorrhagic stroke) and 10,000 controls. The odds ratios of TIA, ischemic stroke, and hemorrhagic stroke among women currently using HT were 1.48 (95% CI, 1.17-1.87), 1.12 (95% CI, 0.78-1.59) and 1.21 (95% CI, 0.76-1.93), respectively, compared to never users. The overall risk estimate for having a cerebrovascular event was 1.34 (95% CI, 1.11-1.61). The risk of TIA was greater (1.96) among women using high doses of estrogen (95% CI, 1.34-2.87). Overall, a small increased risk of stroke associated with HT use of comparable magnitude to the one observed in recent clinical trials was found. The increased risk was more apparent for TIA than for stroke and was greater at higher doses.


Hypercholesterolemia increases vasospasm resulting from basilar artery subarachnoid hemorrhage in rabbits which is attenuated by Vitamin E.
Aneurysm rupture results in subarachnoid hemorrhage (SAH) with subsequent vasospasm in the cerebral and cerebellar major arteries. In recent years, there has been increasing evidence that hypercholesterolemia plays a role in the pathology of SAH. It is known that hypercholesterolemia is one of the major risk factors for the development of atherosclerosis. Among the factors that have been found to retard the development of atherosclerosis is the intake of a sufficient amount of Vitamin E. An inverse association between serum Vitamin E and coronary heart disease mortality has been demonstrated in epidemiologic studies. Therefore, we tested, in an established model of enhanced cholesterol feed in rabbits, the effects of hypercholesterolemia on vasospasm after SAH by using computed tomography (CT) angiograms of the rabbit basilar artery; in addition, we tested the effects of Vitamin E on these conditions, which have not been studied up to now. In this study rabbits were divided into 3 major groups: control, cholesterol fed, and cholesterol + Vitamin E fed. Hypercholesterolemia was induced by a 2% cholesterol-containing diet. Three rabbit groups were fed rabbit diet; one group was fed a diet that also contained 2% cholesterol and another group was fed a diet containing 2% cholesterol and they received i.m. injections of 50 mg/kg of Vitamin E. After 8 weeks, SAH was induced by the double-hemorrhage method and distilled water was injected into cisterna magna. Blood was taken to measure serum cholesterol and Vitamin E levels. Basilar artery samples were taken for microscopic examination. CT angiography and measurement of basilar artery diameter were performed at days 0 and 3 after SAH. Two percent cholesterol diet supplementation for 8 weeks resulted in a significant increase in serum cholesterol levels. Light microscopic analysis of basilar artery of hypercholesterolemic rabbits showed disturbances in the subendothelial and medial layers, degeneration of elastic fibers in the medial layer from endothelial cell desquamation, and a reduction of waves in the endothelial layer. However, the cholesterol + Vitamin E group did not exhibit these changes. The mean diameter of the basilar artery after SAH induction in the cholesterol-treated group was decreased 47% compared with the mean diameter of the control group. This value was less affected in cholesterol + Vitamin E-treated rabbits, which decreased 18% compared with the mean diameter of the control group. Hypercholesterolemia-related changes in the basilar artery aggravate vasospasm after SAH. Adding Vitamin E to cholesterol-treated rabbits decreased the degree of vasospasm following SAH in the rabbit basilar artery SAH model. We suggest that Vitamin E supplements and a low cholesterol diet may potentially diminish SAH complicated by vasospasm in high-risk patients.


High-throughput screening for industrial enzyme production hosts by droplet microfluidics.
A high-throughput method for single cell screening by microfluidic droplet sorting is applied to a whole-genome mutated yeast cell library yielding improved production hosts of secreted industrial enzymes. The sorting method is validated by enriching a yeast strain 14 times based on its α-amylase production, close to the theoretical maximum enrichment. Furthermore, a 10(5) member yeast cell library is screened yielding a clone with a more than 2-fold increase in α-amylase production. The increase in enzyme production results from an improvement of the cellular functions of the production host in contrast to previous droplet-based directed evolution that has focused on improving enzyme protein structure. In the workflow presented, enzyme producing single cells are encapsulated in 20 pL droplets with a fluorogenic reporter substrate. The coupling of a desired phenotype (secreted enzyme concentration) with the genotype (contained in the cell) inside a droplet enables selection of single cells with improved enzyme production capacity by droplet sorting. The platform has a throughput over 300 times higher than that of the current industry standard, an automated microtiter plate screening system. At the same time, reagent consumption for a screening experiment is decreased a million fold, greatly reducing the costs of evolutionary engineering of production strains.


Relationships among multifocal electroretinogram amplitude, visual field sensitivity, and SD-OCT receptor layer thicknesses in patients with retinitis pigmentosa.
To compare local functional measures, the multifocal electroretinogram (mfERG) and visual field sensitivity, with a local structural measure, spectral domain (SD) optical coherence tomography (OCT), of receptor damage in patients with retinitis pigmentosa (RP). MfERGs, visual fields, and SD-OCT scans were obtained from 10 patients with RP, ranging in age from 23 to 59 years. Average amplitudes, average linear sensitivities, and average layer thicknesses were measured from within the central 3° and from three concentric annuli located between 3° and 8°, 8° and 15°, and 15° and 24°. A computer program aided manual segmentation and calculated OCT thickness in the scans. Within each patient with RP, mfERG amplitude for each circle/annulus was highly correlated with corresponding layer thicknesses in the outer retina (r = 0.88 to 0.99), but not at all correlated with thickness of the inner nuclear layer or total retina. Across all ring eccentricities, relative mfERG amplitude and relative visual field sensitivity were correlated with relative SD-OCT outer retinal thickness. In patients with RP, preserved cone photoreceptor function measured by mfERG amplitude and visual field sensitivity correlate well with the remaining thickness of the photoreceptor layer. All three measures show comparable relative loss beyond 3° eccentricity. In the fovea, SD-OCT outer retina thickness showed less relative loss than either mfERG or visual field sensitivity.


Three cadherin alleles associated with resistance to Bacillus thuringiensis in pink bollworm.
Evolution of resistance by pests is the main threat to long-term insect control by transgenic crops that produce Bacillus thuringiensis (Bt) toxins. Because inheritance of resistance to the Bt toxins in transgenic crops is typically recessive, DNA-based screening for resistance alleles in heterozygotes is potentially much more efficient than detection of resistant homozygotes with bioassays. Such screening, however, requires knowledge of the resistance alleles in field populations of pests that are associated with survival on Bt crops. Here we report that field populations of pink bollworm (Pectinophora gossypiella), a major cotton pest, harbored three mutant alleles of a cadherin-encoding gene linked with resistance to Bt toxin Cry1Ac and survival on transgenic Bt cotton. Each of the three resistance alleles has a deletion expected to eliminate at least eight amino acids upstream of the putative toxin-binding region of the cadherin protein. Larvae with two resistance alleles in any combination were resistant, whereas those with one or none were susceptible to Cry1Ac. Together with previous evidence, the results reported here identify the cadherin gene as a leading target for DNA-based screening of resistance to Bt crops in lepidopteran pests.


Pre-eclampsia is associated with, and preceded by, hypertriglyceridaemia: a meta-analysis.
Elevated triglycerides are a feature of the metabolic syndrome, maternal obesity, maternal vasculitis (i.e. systemic lupus erythematosus) and diabetes mellitus. These conditions are all known risk factors for pre-eclampsia. Hypertriglyceridaemia therefore may be associated with pre-eclampsia and indeed this may precede the presence of overt disease. In this study we determine the association between hypertriglyceridaemia and pre-eclampsia in pregnant women. We searched MEDLINE, EMBASE, Web of Science, Excerpta Medica Database, ISI Web of Knowledge, Cumulative Index to Nursing and Allied Health Literature, Cochrane Library from inception until June 2012 and reference lists of relevant studies. Two reviewers independently selected studies on pregnant women where triglycerides were measured and women were followed up until the development of pre-eclampsia or selected on the basis of presence of pre-eclampsia and compared with controls. We collected and meta-analysed the weighted mean differences (WMDs) of triglyceride levels from individual studies using a random effects model. We found strong evidence from meta-analysis of 24 case-control studies (2720 women) that pre-eclampsia is associated with higher levels of serum triglycerides (WMD 0.78 mmol/l, 95% confidence interval 0.6-0.96, P < 0.00001). This finding is also confirmed in five cohort studies, that recruited 3147 women in the second trimester before the onset of pre-eclampsia, which proves that hypertriglyceridaemia precedes the onset of pre-eclampsia (WMD 0.24 mmol/l, 95% confidence interval 0.13-0.34, P < 0.0001). Hypertriglyceridaemia is associated with and precedes the onset of pre-eclampsia. Further research should focus on defining the prognostic accuracy of this test to identify women at risk and the beneficial effect of triglyceride-lowering therapies in pregnancy.


Update on disease-modifying therapies for multiple sclerosis.
Multiple sclerosis (MS) is an autoimmune, demyelinating disease of the central nervous system (CNS). It predominantly affects young women and is one of the most common causes of disability in young adults. MS is characterized by formation of white matter lesions in the CNS as a result of inflammation, demyelination, and axonal loss. Treatment has been a focus of neurological research for over 60 years. A number of disease-modifying therapies (DMTs) have become available making MS a treatable disease. These compounds target the inflammatory response in MS. They work by decreasing the chances of relapse, decreasing the chances of new lesion formation seen on MRI of the CNS and slowing the accumulation of disability. The first drugs for MS to be available were interferon-β and glatiramer acetate. These work by modulating the inflammatory response via different mechanisms that are briefly discussed. Newer agents have since become available and have significantly changed the dynamics of MS treatment. These include fingolimod, dimethyl fumarate and teriflunomide, which are oral agents. Other second-line and third-line Food and Drug Administration (FDA) approved medications include natalizumab and alemtuzumab. Natalizumab is considered one of the most potent treatments for relapse prevention. However, the high risk of progressive multifocal leukoencephalopathy (PML), which is caused by JC virus infection in the brain, tempers the more widespread use of this agent; nevertheless, JC virus antibody tests have helped to stratify the risk of PML. Alemtuzumab, which also has a considerable side effect profile, is likewise highly efficacious. Ocrelizumab, a monoclonal antibody to CD20 on B cells, is a highly effective agent for MS that is likely to be approved soon by the FDA. MS is a major contributor to healthcare costs and it is critical that healthcare providers be aware of the availability and benefits of DMTs. It is imperative that prompt and adequate treatment be established on diagnosis. Changes in therapy should be considered when there is evidence of disease activity as well as accumulation of disability or safety or tolerability concerns.


Two dimensional mapping of the perfusion of the retina and optic nerve head.
To present a new non-invasive method of performing a high definition topography of perfused vessels of the retina and the optic nerve head with simultaneous evaluation of blood flow. By a combination of a laser Doppler flowmeter with a scanning laser system the perfusion of the retina and the optic nerve head is visualised. The principles of measuring blood flow by laser Doppler flowmetry are based on the optical Doppler effect: laser light scattered by a moving particle is shifted in frequency by an amount delta f. Our data acquisition and evaluation system is a modified laser scanning tomograph. The technical data are retinal area of measurement 2.7 mm x 0.7 mm, 10 degrees field with 256 points x 64 lines, measurement accuracy 10 microns, wavelength 670 nm and 790 nm, light power 100 microW and 200 microW, data acquisition time 2.048 s. Every line is scanned 128 times by a line sampling rate of 4000 Hz. By performing a discrete fast Fourier transformation over 128 intensities of each retinal point the laser Doppler shift is calculated for each retinal point. With these data a two dimensional map with 256 x 64 points of the retinal perfusion is created. The brightness of the pixel is coded by the value of the Doppler shift. Offline capillary blood flow is estimated in arbitrary units according to the theory of laser Doppler flowmetry in every region of interest of the perfusion picture. We estimated the reliability and the validity of the method. Retinal blood flow was measured by scanning laser Doppler flowmetry (SLDF) while varying intraocular pressure by a suction cup of three healthy volunteers. Measurements of retinal blood flow performed in 47 eyes by the presented method (SLDF) were correlated with data gained by a commercially available laser Doppler flowmeter. Perfusion pictures of the superficial retinal layer and of deep prelaminar layers in the optic nerve head are presented. The reliability coefficients r1 of 'flow', 'volume', and 'velocity' were 0.84, 0.85, and 0.84 respectively. We found a significant linear relation between SLDF flow and the ocular perfusion pressure (r = 0.84, p < 0.001). Comparative measurements of the retinal blood flow by SLDF and a commercially available laser Doppler flowmeter showed a linear and significant relation (flow r = 0.6, p < 0.0001, volume r = 0.4, p < 0.01). Capillaries of the retinal superficial vasculature or deep ciliary sourced capillaries of the optic nerve head became visible with a high resolution by the confocal technique dependent on the focus. Offline, the blood flow variables of areas of 100 microns x 100 microns were calculated. SLDF enables the visualisation of perfused capillaries and vessels of the retina and the optic nerve head in high resolution by two dimensional mapping of perfusion variables which are encoded by the Doppler signal. This method achieves simultaneously qualitative and quantitative evaluation of capillary blood flow of distinct areas of the capillary meshwork.


Preoperative effects of mifepristone for dilation and evacuation after 19 weeks of gestation: a randomised controlled trial.
To evaluate mifepristone as an adjunct to, or replacement for, osmotic dilators for cervical preparation in surgical abortion after 19 weeks of gestation. Site-stratified, double-blinded randomised controlled trial. Two tertiary care teaching hospitals. Women undergoing dilation and evacuation at 19-236/7 weeks of gestation from November 2013 through November 2015. Participants were randomised to receive (1) mifepristone alone (n = 27), (2) osmotic dilators with mifepristone (n = 27) or (3) osmotic dilators with placebo (n = 21) with all receiving pre-procedure misoprostol. Operative time, preoperative cervical dilation and complications. We enrolled 75 participants; mean gestation 21 weeks. Pre-procedure cervical dilation was ≥3 cm in 4, 52, and 57% of participants in groups 1, 2, and 3, respectively (P < 0.005). Mifepristone with misoprostol for cervical preparation resulted in longer procedure times compared with osmotic dilators, with median total procedure times of (1) 18.5 (8-52), (2) 12 (7-25), and (3) 13 (6-26) minutes (P ≤ 0.005). Excluding time required for manual dilation, procedure times were similar: median times from dilation complete to evacuation complete were (1) 10.5 (4-23), (2) 8.5 (5-24), and (3) 10 (4-20) minutes (P = 0.10). Complications occurred in seven cases, six with trainees and one with an attending physician (P = 0.03), with difference by study group not reaching statistical significance (P = 0.12). Elimination of osmotic dilators has the potential to decrease burden and opportunity cost of cervical preparation. The longer procedure time, related to manual dilation, is offset by decreasing dilator-related preoperative time and discomfort. Provider experience may impact risk when eliminating dilators. Mifepristone and misoprostol for cervical preparation prior to D&E has potential to reduce barriers to care.


Fast Robust Dejitter and Interslice Discontinuity Removal in MRI Phase Acquisitions: Application to Magnetic Resonance Elastography.
MRI phase contrast imaging methods that assemble slice-wise acquisitions into volumes can contain interslice phase discontinuities (IPDs) over the course of the scan from sources, including unavoidable physiological activity. In magnetic resonance elastography (MRE), this can alter wavelength and tissue stiffness estimates, invalidating the analysis. We first model this behavior as jitter along the z-axis of the phase of 3D complex-valued wave volumes. A two-step image processing pipeline is then proposed that removes IPDs. First, constant slicewise phase shift is removed with a novel, non-convex dejittering algorithm. Then, regional physiological noise artifacts are removed with novel filtering of 3D wavelet coefficients. Calibration of two pipeline coefficients, the dejitter parameter α and the wavelet band high-pass coefficient ωc , was first performed on a finite-element method brain phantom. A comparative investigation was then performed, on a cohort of 48 brain acquisitions, of four approaches to IPDs: 1) the proposed method; 2) a "control" condition of neglect of IPDs; 3) an anisotropic wavelet-based method; and 4) a method of in-plane (2D) processing. The present method showed medians of [Formula: see text] Pa for a multifrequency wave inversion centered at 40 Hz which was within 6% of methods 3) and 4), while neglect produced [Formula: see text] estimates a mean of 17% lower. The proposed method reduced the value range of the cohort against methods 3) and 4) by 29% and 31%, respectively. Such reduction in variance enhances the ability of brain MRE to predict subtler physiological changes. Our theoretical approach further enables more powerful applications of fundamental findings in noise and denoising to MRE.


AtlA Mediates Extracellular DNA Release, Which Contributes to Streptococcus mutans Biofilm Formation in an Experimental Rat Model of Infective Endocarditis.
Host factors, such as platelets, have been shown to enhance biofilm formation by oral commensal streptococci, inducing infective endocarditis (IE), but how bacterial components contribute to biofilm formation in vivo is still not clear. We demonstrated previously that an isogenic mutant strain of Streptococcus mutans deficient in autolysin AtlA (ΔatlA) showed a reduced ability to cause vegetation in a rat model of bacterial endocarditis. However, the role of AtlA in bacterial biofilm formation is unclear. In this study, confocal laser scanning microscopy analysis showed that extracellular DNA (eDNA) was embedded in S. mutans GS5 floes during biofilm formation on damaged heart valves, but an ΔatlA strain could not form bacterial aggregates. Semiquantification of eDNA by PCR with bacterial 16S rRNA primers demonstrated that the ΔatlA mutant strain produced dramatically less eDNA than the wild type. Similar results were observed with in vitro biofilm models. The addition of polyanethol sulfonate, a chemical lysis inhibitor, revealed that eDNA release mediated by bacterial cell lysis is required for biofilm initiation and maturation in the wild-type strain. Supplementation of cultures with calcium ions reduced wild-type growth but increased eDNA release and biofilm mass. The effect of calcium ions on biofilm formation was abolished in ΔatlA cultures and by the addition of polyanethol sulfonate. The VicK sensor, but not CiaH, was found to be required for the induction of eDNA release or the stimulation of biofilm formation by calcium ions. These data suggest that calcium ion-regulated AtlA maturation mediates the release of eDNA by S. mutans, which contributes to biofilm formation in infective endocarditis.


The retinoblastoma binding protein RBP2 is an H3K4 demethylase.
Changes in histone methylation status regulate chromatin structure and DNA-dependent processes such as transcription. Recent studies indicate that, analogous to other histone modifications, histone methylation is reversible. Retinoblastoma binding protein 2 (RBP2), a nuclear protein implicated in the regulation of transcription and differentiation by the retinoblastoma tumor suppressor protein, contains a JmjC domain recently defined as a histone demethylase signature motif. Here we report that RBP2 is a demethylase that specifically catalyzes demethylation on H3K4, whose methylation is normally associated with transcriptionally active genes. RBP2-/- mouse cells displayed enhanced transcription of certain cytokine genes, which, in the case of SDF1, was associated with increased H3K4 trimethylation. Furthermore, RBP2 specifically demethylated H3K4 in biochemical and cell-based assays. These studies provide mechanistic insights into transcriptional regulation by RBP2 and provide the first example of a mammalian enzyme capable of erasing trimethylated H3K4.


Failure to detect immune deficiency in rats after prenatal or early postnatal irradiation.
We have looked for medium-term sequelae in the immune system of rats that had been X-irradiated (0-2 Gy whole-body irradiation) during prenatal or early postnatal life. At an age of 8 weeks the histology of the spleen was normal, and so was the distribution of B and T lymphocytes. The serum immunoglobulin levels were not significantly altered, even when the different isotypes were considered. At an age of 10 weeks, the rats were immunized with a T-dependent or a T-independent dinitrophenylated-carrier antigen. Normal levels of specific antibodies were generated in all groups of animals injected with the T-independent antigen. The T-dependent response, in contrast, was higher in animals irradiated between day 6 and day 20 of gestation (but not in rats irradiated early after birth). This increase, however, was significant only for the IgM and IgG1 responses of some irradiated groups. Thus no medium-term immunodeficiency could be documented with the methods used. The alteration in a T-dependent response, however, points to a radiosensitive T regulatory mechanism.


Clinicopathological correlates and prognostic significance of KRAS mutation status in a pooled prospective cohort of epithelial ovarian cancer.
Activating KRAS mutations are common in ovarian carcinomas of low histological grade, less advanced clinical stage and mucinous histological subtype, and form part of the distinct molecular alterations associated with type I tumors in the dualistic model of ovarian carcinogenesis. Here, we investigated the occurrence, clinicopathological correlates and prognostic significance of specific KRAS mutations in tumours from 153 epithelial ovarian cancer (EOC) cases from a pooled, prospective cohort. KRAS codon 12,13 and 61 mutations were analysed by pyrosequencing in tumours from 163 incident EOC cases in the Malmö Diet and Cancer Study and Malmö Preventive Project. Associations of mutational status with clinicopathological and molecular characteristics were assessed by Pearson Chi Square test. Ovarian cancer-specific survival (OCSS) according to mutational status was explored by Kaplan-Meier analysis and Cox proportional hazards modelling. KRAS-mutation status was also analysed in 28 concomitantly sampled benign-appearing fallopian tubes. Seventeen (11.1%) EOC cases harboured mutations in the KRAS gene, all but one in codon 12, and one in codon 13. No KRAS mutations were found in codon 61 and all examined fallopian tubes were KRAS wild-type. KRAS mutation was significantly associated with lower grade (p = 0.001), mucinous histological subtype (p = < 0.001) and progesterone receptor expression (p = 0.035). Kaplan-Meier analysis revealed a significantly improved OCSS for patients with KRAS-mutated compared to KRAS wild-type tumours (p = 0.015). These associations were confirmed in unadjusted Cox regression analysis (HR = 2.51; 95% CI 1.17-5.42) but did not remain significant after adjustment for age, grade and clinical stage. The beneficial prognostic impact of KRAS mutation was ony evident in tumours of low-intermediate differentiation grade (p = 0.023), and in a less advanced clinical stage (p = 0.014). Moreover, KRAS mutation was associated with a significantly improved OCSS in the subgroup of endometroid carcinomas (p = 0.012). The results from this study confirm previously demonstrated associations of KRAS mutations with well-differentiated and mucinous ovarian carcinomas. Moreover, KRAS-mutated tumours had a significantly improved survival in unadjusted, but not adjusted, analysis. A finding that merits further study is the significant prognostic impact of KRAS mutation in endometroid carcinomas, potentially indicating that response to Ras/Raf/MEK/ERK-targeting therapies may differ by histological subtype. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1788330379100147.


Effect of Preoperative Inspiratory Muscle Training on Alveolar-Arterial Oxygen Gradients After Coronary Artery Bypass Surgery.
Gas exchange abnormalities and inspiratory muscle dysfunction are common postoperative pulmonary complications after cardiac surgery. The aim of this study was to investigate the changes in the alveolar arterial oxygen gradients and inspiratory muscle power after preoperative inspiratory muscle training (IMT) followed by postoperative IMT and early mobilization after coronary artery bypass graft surgery. Patients were randomly assigned to the intervention group (n = 20) or the usual care group (n = 20). The intervention group received preoperative IMT by a threshold load inspiratory muscle trainer and education followed by postoperative IMT and early mobilization. The usual care group received preoperative education and postoperative routine physical therapy care. Alveolar-arterial oxygen gradients and oxygen saturation were measured before induction of anesthesia and postoperatively immediately after extubation, at 24 hours, 48 hours, and on day 8 postsurgery. Inspiratory muscle power was measured before surgery, after 48 hours, and on day 8. Forty male patients underwent coronary artery bypass graft surgery. The intervention group showed significant (P < .05) improvement of alveolar arterial-oxygen gradients and inspiratory muscle power at all measurement points. The mean of inspiratory muscle power at discharge was significantly higher than at baseline but not statistically significantly different from the preoperative measurement. Pre- and postoperative IMT by the threshold load inspiratory muscle trainer resulted in improvement of inspiratory muscle power, alveolar-arterial gradient, and oxygen saturation.


Organic amendment additions to rangelands: A meta-analysis of multiple ecosystem outcomes.
Interest in land application of organic amendments-such as biosolids, composts, and manures-is growing due to their potential to increase soil carbon and help mitigate climate change, as well as to support soil health and regenerative agriculture. While organic amendments are predominantly applied to croplands, their application is increasingly proposed on relatively arid rangelands that do not typically receive fertilizers or other inputs, creating unique concerns for outcomes such as native plant diversity and water quality. To maximize environmental benefits and minimize potential harms, we must understand how soil, water, and plant communities respond to particular amendments and site conditions. We conducted a global meta-analysis of 92 studies in which organic amendments had been added to arid, semiarid, or Mediterranean rangelands. We found that organic amendments, on average, provide some environmental benefits (increased soil carbon, soil water holding capacity, aboveground net primary productivity, and plant tissue nitrogen; decreased runoff quantity), as well as some environmental harms (increased concentrations of soil lead, runoff nitrate, and runoff phosphorus; increased soil CO2 emissions). Published data were inadequate to fully assess impacts to native plant communities. In our models, adding higher amounts of amendment benefitted four outcomes and harmed two outcomes, whereas adding amendments with higher nitrogen concentrations benefitted two outcomes and harmed four outcomes. This suggests that trade-offs among outcomes are inevitable; however, applying low-N amendments was consistent with both maximizing benefits and minimizing harms. Short study time frames (median 1-2 years), limited geographic scope, and, for some outcomes, few published studies limit longer-term inferences from these models. Nevertheless, they provide a starting point to develop site-specific amendment application strategies aimed toward realizing the potential of this practice to contribute to climate change mitigation while minimizing negative impacts on other environmental goals.


[Regional differences in adults' tea drinking pattern: findings from China Kadoorie Biobank study in 10 areas in China].
To describe the gender and regional differences in adults' tea drinking pattern found in China Kadoorie Biobank (CKB) survey, involving half million adults, in 10 areas in China. The baseline survey of CKB was conducted in 5 urban areas and 5 rural areas in China during 2004-2008. The information on tea drinking of 512 891 adults aged 30-79 years was collected and analyzed. The overall prevalence of regular tea drinking (i.e., weekly) was 50.7% in males and 21.3% in females. Among 10 areas, Hunan reported the highest regular tea drinking prevalence (74.3% in males, 76.6% in females), while Henan reported the lowest regular tea drinking prevalence (5.5% in males, 1.0% in females). Most regular tea drinkers consumed green tea except those from Zhejiang and Haikou. Hunan reported the highest tea consumption per time (3.8 g in males, 3.1 g in females), per day (6.2 g in males, 4.1 g in females) and per week (38.4 g in males, 25.0 g in females). The adults' tea drinking pattern i.e. frequency, type, amount of tea and preferred concentration of tea water, varied greatly among the 10 regions covered by CKB study.


Neural systems for vocal learning in birds and humans: a synopsis.
I present here a synopsis on a hypothesis that I derived on the similarities and differences of vocal learning systems in vocal learning birds for learned song and in humans for spoken language. This hypothesis states that vocal learning birds-songbirds, parrots, and hummingbirds-and humans have comparable specialized forebrain regions that are not found in their close vocal non-learning relatives. In vocal learning birds, these forebrain regions appear to be divided into two sub-pathways, a vocal motor pathway mainly used to produce learned vocalizations and a pallial-basal-ganglia-thalamic loop mainly used to learn and modify the vocalizations. I propose that humans have analogous forebrain pathways within and adjacent to the motor and pre-motor cortices, respectively, used to produce and learn speech. Recent advances have supported the existence of the seven cerebral vocal nuclei in the vocal learning birds and the proposed brain regions in humans. The results in birds suggest that the reason why the forebrain regions are similar across distantly related vocal learners is that the vocal pathways may have evolved out of a pre-existing motor pathway that predates the ancient split from the common ancestor of birds and mammals. Although this hypothesis will require the development of novel technologies to be fully tested, the existing evidence suggest that there are strong genetic constraints on how vocal learning neural systems can evolve.


Nickel-catalyzed cross-coupling of aldehydes with aryl halides via hydrazone intermediates.
Traditional cross-couplings require stoichiometric organometallic reagents. A novel nickel-catalyzed cross-coupling reaction between aldehydes and aryl halides via hydrazone intermediates has been developed, merging the Wolff-Kishner reduction and the classical cross-coupling reactions. Aromatic aldehydes, aryl iodides and aryl bromides are especially effective in this new cross-coupling chemistry.


Hyperuricemia induces a primary renal arteriolopathy in rats by a blood pressure-independent mechanism.
Hyperuricemia is associated with hypertension and vascular disease, but whether this represents a causal relationship or an epiphenomenon remains unknown. We recently reported a model of mild hyperuricemia in rats that results in increased blood pressure and mild renal fibrosis. In this study, we examined the effect of hyperuricemia on the renal vasculature. Rats fed 2% oxonic acid and a low-salt diet for 7 wk developed mild hyperuricemia (1.8 vs. 1.4 mg/dl, P < 0.05), hypertension [147 vs. 127 mmHg systolic blood pressure (SBP), P < 0.05], and afferent arteriolar thickening, with a 35% increase in medial area (P < 0.05). Allopurinol or benziodarone prevented the hyperuricemia, hypertension, and arteriolopathy. Hydrochlorothiazide treatment did not prevent the hyperuricemia or arteriolopathy despite controlling blood pressure. In contrast, the arteriolopathy and hypertension were prevented by both enalapril and losartan. Uric acid also directly stimulated vascular smooth muscle cell proliferation in vitro, and this was partially inhibited by losartan. Thus hyperuricemia induces a renal arteriolopathy in rats that is blood pressure independent and involves the renin-angiotensin system.


Malaria infection and severe disease risks in Africa.
The relationship between community prevalence of Plasmodium falciparum and the burden of severe, life-threatening disease remains poorly defined. To examine the three most common severe malaria phenotypes from catchment populations across East Africa, we assembled a dataset of 6506 hospital admissions for malaria in children aged 3 months to 9 years from 2006 to 2020. Admissions were paired with data from community parasite infection surveys. A Bayesian procedure was used to calibrate uncertainties in exposure (parasite prevalence) and outcomes (severe malaria phenotypes). Each 25% increase in prevalence conferred a doubling of severe malaria admission rates. Severe malaria remains a burden predominantly among young children (3 to 59 months) across a wide range of community prevalence typical of East Africa. This study offers a quantitative framework for linking malaria parasite prevalence and severe disease outcomes in children.


The Relationship Between Zinc and Quality of Life in Patients With Upper GI Cancer on Chemotherapy.
This is a pilot study aimed at evaluating the prevalence of zinc deficiency and how zinc levels affect the quality of life (QOL) of patients with upper gastrointestinal (GI) cancers receiving systemic chemotherapy. The data collection was completed on 40 patients. Although the primary objective of a positive prevalence of zinc deficiency in upper GI cancer patients at diagnosis and after receiving chemotherapy is not statistically significant, we found a statistically significant association between zinc level and certain QOL factors. There is a significantly positive association with satisfaction of social contact at baseline only, sexual pleasure at baseline and at 2 months, QOL at baseline only, and troublesome sweating at baseline, and from baseline to 2 months corresponding with change in other skin problems. Conversely, there is a significantly negative association corresponding changes in enjoyment of physical activities, how the patient usually feels, sexual pleasure, the way in which the patient approaches food, QOL, rashes on the face, and other skin problems. Neutropenia grades were reflective of decreased zinc at baseline but did not show decreased zinc correlating with a weakened immune system.


Improving syllable identification by a preprocessing method reducing overlap-masking in reverberant environments.
Overlap-masking degrades speech intelligibility in reverberation [R. H. Bolt and A. D. MacDonald, J. Acoust. Soc. Am. 21(6), 577-580 (1949)]. To reduce the effect of this degradation, steady-state suppression has been proposed as a preprocessing technique [Arai et al., Proc. Autumn Meet. Acoust. Soc. Jpn., 2001; Acoust. Sci. Tech. 23(8), 229-232 (2002)]. This technique automatically suppresses steady-state portions of speech that have more energy but are less crucial for speech perception. The present paper explores the effect of steady-state suppression on syllable identification preceded by /a/ under various reverberant conditions. In each of two perception experiments, stimuli were presented to 22 subjects with normal hearing. The stimuli consisted of mono-syllables in a carrier phrase with and without steady-state suppression and were presented under different reverberant conditions using artificial impulse responses. The results indicate that steady-state suppression statistically improves consonant identification for reverberation times of 0.7 to 1.2 s. Analysis of confusion matrices shows that identification of voiced consonants, stop and nasal consonants, and bilabial, alveolar, and velar consonants were especially improved by steady-state suppression. The steady-state suppression is demonstrated to be an effective preprocessing method for improving syllable identification by reducing the effect of overlap-masking under specific reverberant conditions.


Angiotensin II stimulates calcineurin activity in proximal tubule epithelia through AT-1 receptor-mediated tyrosine phosphorylation of the PLC-gamma1 isoform.
Angiotensin II (AngII) contributes to the maintenance of extracellular fluid volume by regulating sodium transport in the nephron. In nonepithelial cells, activation of phospholipase C (PLC) by AT-1 receptors stimulates the generation of 1,4,5-trisphosphate (IP(3)) and the release of intracellular calcium. Calcineurin, a serine-threonine phosphatase, is activated by calcium and calmodulin, and both PLC and calcineurin have been linked to sodium transport in the proximal tubule. An examination of whether AngII activates calcineurin in a model of proximal tubule epithelia (LLC-PK1 cells) was performed; AngII increased calcineurin activity within 30 s. An examination of whether AngII activates PLC in proximal tubule epithelia was also performed after first showing that all three families of PLC isoforms are present in LLC-PK1 cells. Application of AngII increased IP(3) generation by 60% within 15 s, which coincided with AngII-induced tyrosine phosphorylation of the PLC-gamma1 isoform also observed at 15 s. AngII-induced tyrosine phosphorylation was blocked by the AT-1 receptor antagonist, Losartan. Subsequently, an inhibitor of tyrosine phosphorylation blocked the AngII-induced activation of calcineurin, as did coincubation with an inhibitor of PLC activity and with an antagonist of the AT-1 receptor. It is therefore concluded that AngII stimulates calcineurin phosphatase activity in proximal tubule epithelial cells through a mechanism involving AT-1 receptor-mediated tyrosine phosphorylation of the PLC isoform.


Polyelectrolyte multilayers under compression: concurrent osmotic stress and colloidal probe atomic force microscopy.
Colloidal interactions have been characterised using both osmotic stress and surface forces. Here these methods are employed concurrently to measure the interaction forces of polyelectrolyte multilayers that when cross-linked form a dextran impermeable membrane. The force data, corrected for the thickness of the polyelectrolyte multilayer film, has been expressed as pressure versus separation enabling the interaction from osmotic stress measurements to be compared to the measured interaction from the colloid probe technique. The combined technique is valuable in evaluating the interaction forces associated with compression of polymer films at different rates and over a wide range of pressure and demonstrates features that are not revealed when just one technique is employed. The combination of the techniques allows both attractive forces and strongly repulsive forces to be measured and shows that the measured repulsion is greater in the force data than in the osmotic data. This is due to insufficient equilibration time in the AFM measurements, even at the slowest approach rates available, indicating that AFM force measurements between polyelectrolytes will always contain a dynamic component. That is we demonstrate that colloid probe measurements between polymer surfaces overestimate the equilibrium repulsive interaction due to the rate at which the measurement is performed.


Recent developments in the laboratory diagnosis of chlamydial infections.
There are two main approaches to diagnosing infections by Chlamydia and Chlamydophila spp. in mammals and birds. The first involves the direct detection of the agent in tissue or swab samples, while the second involves the serological screening of blood samples for the presence of anti-chlamydial antibodies. Ultimately, the test that is used is dependent on the types of samples that are submitted to the diagnostic laboratory for analysis. The present paper gives an overview on methodologies and technologies used currently in diagnosis of chlamydial infections with emphasis on recently developed tests. The performance characteristics of individual methods, such as the detection of antigen in smears and in pathological samples, the isolation of the pathogen, various antibody detection tests and DNA-based methods utilising conventional and real-time PCR, as well as DNA microarray technology are assessed, and specific advantages and drawbacks are discussed. Further, a combination of a specific real-time PCR assay and a microarray test for chlamydiae is proposed as an alternative reference standard to isolation by cell culture.


Combination chemotherapy and systemic irradiation consolidation for poor prognosis breast cancer.
Seventy patients with poor prognosis, metastatic breast cancer were treated with FUVAC induction chemotherapy (5-fluorouracil, vinblastine, Adriamycin [doxorubicin] and cyclophosphamide). Consolidation therapy was given to 30 of 48 responders (63%), of whom 23 received sequential hemibody irradiation (HBI) at 8 cGy, corrected in the upper half for lung transmission. Seven received high dose cyclophosphamide and total body irradiation (TBI) with subsequent infusion of stored, cryopreserved autologous bone marrow. The response rate to induction therapy was 71% (complete [CR] in 21%). The median survival for all patients entered in this study is 12 months. With consolidation, one CR patient who received cyclophosphamide and TBI is disease free at 20+ months, off all treatment, while HBI did not produce longterm remissions. Of 17 partial response (PR) patients, two of 12 improved to CR with HBI, and one of five improved with cyclophosphamide plus TBI, but all ultimately relapsed. The main toxicity of sequential HBI was myelosuppression, with prolonged thrombocytopenia in 13%; only one case of radiation pneumonitis occurred (3%). Cyclophosphamide and TBI produced temporary, reversible marrow aplasia without other major toxicity. We recommend further investigation of Cytoxan (Bristol Myers Oncology Division, Evansville, IN) and TBI for breast cancer patients in remission after chemotherapy.


Cross-species functional diversity within the PIN auxin efflux protein family.
In Arabidopsis, development during flowering is coordinated by transport of the hormone auxin mediated by polar-localized PIN-FORMED1 (AtPIN1). However Arabidopsis has lost a PIN clade sister to AtPIN1, Sister-of-PIN1 (SoPIN1), which is conserved in flowering plants. We previously proposed that the AtPIN1 organ initiation and vein patterning functions are split between the SoPIN1 and PIN1 clades in grasses. Here we show that in the grass Brachypodium sopin1 mutants have organ initiation defects similar to Arabidopsis atpin1, while loss of PIN1 function in Brachypodium has little effect on organ initiation but alters stem growth. Heterologous expression of Brachypodium SoPIN1 and PIN1b in Arabidopsis provides further evidence of functional specificity. SoPIN1 but not PIN1b can mediate flower formation in null atpin1 mutants, although both can complement a missense allele. The behavior of SoPIN1 and PIN1b in Arabidopsis illustrates how membrane and tissue-level accumulation, transport activity, and interaction contribute to PIN functional specificity.


Reduced fMRI activity predicts relapse in patients recovering from stimulant dependence.
Relapse presents a significant problem for patients recovering from stimulant dependence. Here we examined the hypothesis that patterns of brain function obtained at an early stage of abstinence differentiates patients who later relapse versus those who remain abstinent. Forty-five recently abstinent stimulant-dependent patients were tested using a randomized event-related functional MRI (ER-fMRI) design that was developed in order to replicate a previous ERP study of relapse using a selective attention task, and were then monitored until 6 months of verified abstinence or stimulant use occurred. SPM revealed smaller absolute blood oxygen level-dependent (BOLD) response amplitude in bilateral ventral posterior cingulate and right insular cortex in 23 patients positive for relapse to stimulant use compared with 22 who remained abstinent. ER-fMRI, psychiatric, neuropsychological, demographic, personal and family history of drug use were compared in order to form predictive models. ER-fMRI was found to predict abstinence with higher accuracy than any other single measure obtained in this study. Logistic regression using fMRI amplitude in right posterior cingulate and insular cortex predicted abstinence with 77.8% accuracy, which increased to 89.9% accuracy when history of mania was included. Using 10-fold cross-validation, Bayesian logistic regression and multilayer perceptron algorithms provided the highest accuracy of 84.4%. These results, combined with previous studies, suggest that the functional organization of paralimbic brain regions including ventral anterior and posterior cingulate and right insula are related to patients' ability to maintain abstinence. Novel therapies designed to target these paralimbic regions identified using ER-fMRI may improve treatment outcome.


A genetic variant in the IL-17 promoter is functionally associated with acute graft-versus-host disease after unrelated bone marrow transplantation.
Interleukin IL-17 is a proinflammatory cytokine that has been implicated in the pathogenesis of various autoimmune diseases. The single nucleotide polymorphism (SNP), rs2275913, in the promoter region of the IL-17 gene is associated with susceptibility to ulcerative colitis. When we examined the impact of rs2275913 in a cohort consisting of 438 pairs of patients and their unrelated donors transplanted through the Japan Marrow Donor Program, the donor IL-17 197A allele was found to be associated with a higher risk of acute graft-versus-host disease (GVHD; hazard ratio [HR], 1.46; 95% confidence interval [CI], 1.00 to 2.13; P = 0.05). Next, we investigated the functional relevance of the rs2275913 SNP. In vitro stimulated T cells from healthy individuals possessing the 197A allele produced significantly more IL-17 than those without the 197A allele. In a gene reporter assay, the 197A allele construct induced higher luciferase activity than the 197G allele, and the difference was higher in the presence of T cell receptor activation and was abrogated by cyclosporine treatment. Moreover, the 197A allele displayed a higher affinity for the nuclear factor activated T cells (NFAT), a critical transcription factor involved in IL-17 regulation. These findings substantiate the functional relevance of the rs2275913 polymorphism and indicate that the higher IL-17 secretion by individuals with the 197A allele likely accounts for their increased risk for acute GVHD and certain autoimmune diseases.


Hbo1 Links p53-dependent stress signaling to DNA replication licensing.
Hbo1 is a histone acetyltransferase (HAT) that is required for global histone H4 acetylation, steroid-dependent transcription, and chromatin loading of MCM2-7 during DNA replication licensing. It is the catalytic subunit of protein complexes that include ING and JADE proteins, growth regulatory factors and candidate tumor suppressors. These complexes are thought to act via tumor suppressor p53, but the molecular mechanisms and links between stress signaling and chromatin, are currently unknown. Here, we show that p53 physically interacts with Hbo1 and negatively regulates its HAT activity in vitro and in cells. Two physiological stresses that stabilize p53, hyperosmotic shock and DNA replication fork arrest, also inhibit Hbo1 HAT activity in a p53-dependent manner. Hyperosmotic stress during G(1) phase specifically inhibits the loading of the MCM2-7 complex, providing an example of the chromatin output of this pathway. These results reveal a direct regulatory connection between p53-responsive stress signaling and Hbo1-dependent chromatin pathways.


Toward an in vivo neurochemical profile: quantification of 18 metabolites in short-echo-time (1)H NMR spectra of the rat brain.
Localized in vivo (1)H NMR spectroscopy was performed with 2-ms echo time in the rat brain at 9.4 T. Frequency domain analysis with LCModel showed that the in vivo spectra can be explained by 18 metabolite model solution spectra and a highly structured background, which was attributed to resonances with fivefold shorter in vivo T(1) than metabolites. The high spectral resolution (full width at half maximum approximately 0.025 ppm) and sensitivity (signal-to-noise ratio approximately 45 from a 63-microL volume, 512 scans) was used for the simultaneous measurement of the concentrations of metabolites previously difficult to quantify in (1)H spectra. The strongly represented signals of N-acetylaspartate, glutamate, taurine, myo-inositol, creatine, phosphocreatine, glutamine, and lactate were quantified with Cramér-Rao lower bounds below 4%. Choline groups, phosphorylethanolamine, glucose, glutathione, gamma-aminobutyric acid, N-acetylaspartylglutamate, and alanine were below 13%, whereas aspartate and scyllo-inositol were below 22%. Intra-assay variation was assessed from a time series of 3-min spectra, and the coefficient of variation was similar to the calculated Cramér-Rao lower bounds. Interassay variation was determined from 31 pooled spectra, and the coefficient of variation for total creatine was 7%. Tissue concentrations were found to be in very good agreement with neurochemical data from the literature.


Twin concordance rates for rheumatoid arthritis: results from a nationwide study.
We report the concordance rate for RA in a nationwide study of 91 monozygotic (MZ) and 112 dizygotic (DZ) pairs. Twin pairs were recruited from both a national media campaign and a 2-month prospective inquiry of all UK rheumatologists. Disease status was established following a structured clinical and serological appraisal, together with radiological assessment where necessary. Zygosity was confirmed using DNA fingerprinting. In all, 14 (15.4%) of the MZ and four (3.6%) of the DZ pairs were disease concordant (risk ratio: 4.3 95% CI 1.5 to 12.6). There was no difference in the concordance between the media and clinical derived twins. Further the excess MZ concordance persisted after adjusting for age, age at disease onset, sex and rheumatoid factor status. Analysing the data in relation to the timing of disease onset in the first affected twin showed that subsequent disease risk in the initially unaffected co-twins of the MZ affected probands increased with increasing duration of follow-up. We conclude that the overall MZ concordance at 15% is lower than the 30% figure normally quoted from a study some 30 years ago and sets a ceiling at the potential genetic contribution to disease susceptibility.


Role of the cytoskeleton in signaling networks.
Intracellular signal transduction occurs through cascades of reactions involving dozens of proteins that transmit signals from the cell surface, through a crowded cellular environment filled with organelles and a filamentous cytoskeleton, to specific targets. Numerous signaling molecules are immobilized or transiently bound to the cytoskeleton, yet most models for signaling pathways have no specific role for this mesh, which is often presumed to function primarily as a scaffold that determines cell mechanics but not information flow. We combined analytical tools with several recently established large-scale protein-protein interaction maps for Saccharomyces cerevisiae to quantitatively address the role of the cytoskeleton in intracellular signaling. The results demonstrate that the network of signaling proteins is intimately linked to the cytoskeleton, suggesting that this interconnected filamentous structure plays a crucial and distinct functional role in signal transduction.


Tumor-derived macrophage migration inhibitory factor promotes an autocrine loop that enhances renal cell carcinoma.
The macrophage migration inhibitory factor (MIF) is a hypoxia regulated gene that has a variety of tumorigenic functions. In clear cell renal carcinoma (CCRC), hypoxic signaling is constitutively active because of the frequent loss of function of the von Hippel-Lindau tumor suppressor protein. We therefore sought to assess the expression of MIF in CCRC and its biological functions. We stained tumor tissue microarrays comprising sections of 128 CCRC tumors and found MIF to be moderately or highly expressed in >98%. MIF expression was further found to be dramatically elevated in blood plasma of individuals with CCRC compared with healthy controls, suggesting that measurement of MIF levels in the blood may have utility as a diagnostic marker in CCRC. At a functional level, MIF has been reported to engage the CD74 and CD44 receptors and induce signal transduction. In CCRC cell lines, depletion of MIF, CD74 or CD44 by small hairpin RNA led to a significant reduction in growth rate, and clonogenic survival, coinciding with the degree of knockdown. Interruption of the MIF pathway also decreased tumorigenic potential. Biochemically, we found that in CCRC cells MIF signaling leads to activation of the mitogen-activated protein kinase pathway and to Src phosphorylation, which is critical for regulation of p27. Together, our studies establish MIF as a protumorigenic signaling molecule that functions in an autocrine fashion to promote renal cell carcinoma and may be useful as a minimally invasive marker of disease status.


T cell-derived IL-17 mediates epithelial changes in the airway and drives pulmonary neutrophilia.
Th17 cells are a proinflammatory subset of effector T cells that have been implicated in the pathogenesis of asthma. Their production of the cytokine IL-17 is known to induce local recruitment of neutrophils, but the direct impact of IL-17 on the lung epithelium is poorly understood. In this study, we describe a novel mouse model of spontaneous IL-17-driven lung inflammation that exhibits many similarities to asthma in humans. We have found that STAT3 hyperactivity in T lymphocytes causes an expansion of Th17 cells, which home preferentially to the lungs. IL-17 secretion then leads to neutrophil infiltration and lung epithelial changes, in turn leading to a chronic inflammatory state with increased mucus production and decreased lung function. We used this model to investigate the effects of IL-17 activity on airway epithelium and identified CXCL5 and MIP-2 as important factors in neutrophil recruitment. The neutralization of IL-17 greatly reduces pulmonary neutrophilia, underscoring a key role for IL-17 in promoting chronic airway inflammation. These findings emphasize the role of IL-17 in mediating neutrophil-driven pulmonary inflammation and highlight a new mouse model that may be used for the development of novel therapies targeting Th17 cells in asthma and other chronic pulmonary diseases.


Single fraction radiosurgery, fractionated radiosurgery, and conventional radiotherapy for spinal oligometastasis (SAFFRON): A systematic review and meta-analysis.
To perform a systematic review/meta-analysis of outcomes for patients with spinal metastases treated with stereotactic radiosurgery (SRS) (either single-fraction (SF-SRS) or multiple-fraction (MF-SRS)) or conventional radiotherapy (RT). Thirty-seven studies were identified. Primary outcomes were 1-year local control (LC) and acute/late grade 3-5 toxicities (including vertebral compression fractures (VCF)). Weighted random effects meta-analyses using the DerSimonian and Laird methods and meta-regressions were conducted to characterize and compare effect sizes. Mixed effects regression models were used in dose analyses. A total of 3237 patients with 4911 lesions were included; 43.8%, 19.7%, and 36.5% of lesions received SF-SRS, MF-SRS, or RT, respectively. SF-SRS resulted in improved 1-year LC (92.9% (95% CI: 86.4-97.4%); p = 0.007) compared to RT (81.0% (95% CI: 69.2-90.5%)) with no difference between MF-SRS (82.1%; p = 0.86) and RT. On subgroup analysis of de novo metastases, superior 1-year LC following SF-SRS (95.5% (95% CI: 87.4-99.6%)) was maintained compared to RT (83.6% (95% CI: 70.4-93.5%); p = 0.007). A 4.7% increase in LC was noted for each 10 Gy10 increase in biologically effective dose (BED10, assuming an alpha/beta = 10) with SRS (p < 0.001). No difference in toxicities were found between SF-SRS (0.4%), MF-SRS (0.2%), or RT (0%). Higher VCF rates were noted following SF-SRS (19.5%) vs. MF-SRS (9.6%; p = 0.039)) with no correlation between dose and VCF rates. SF-SRS resulted in superior LC with a roughly 5% LC benefit for every 10 Gy10 increase in BED10 with higher VCF rates compared to MF-SRS. If LC is the goal of treatment, then SRS may be a preferred treatment modality. However, these results are hypothesis-generating, and prospective randomized clinical trials are indicated to definitively address the question of whether SRS results in improved LC compared to RT.


Cancer as the "perfect storm"? A qualitative study of public attitudes to health conditions.
Our aim is to identify important attributes of major diseases that shape how they are perceived by the public. Four focus groups among members of the public were recruited, in March and October 2016, and used semistructured discussion to explore important attributes of cancer, heart disease, stroke, dementia, mental illness, and infectious disease. Common themes were identified by using inductive thematic analysis.Five themes were identified: fear, impact on family and friends, hope, detection, and prevention. Fear of cancer includes not only fear of death but also of aggressive treatments. Loss of dignity is feared in dementia, while infectious disease raises fear of uncontrollable "plague"; in contrast, people with mental illness may themselves be seen as a potential threat. The impact of cancer and its treatment on family and friends was described as intense and all-consuming, even for those not involved directly in caring; with dementia and stroke, the family impact is taking on care, including funding, over the long term with little expectation of improvement. Hope is a major theme in cancer and stroke recovery, linked with the need to take action, often expressed in aggressive language of "fighting," but seen as futile in dementia. Detection difficulties for "silent" cancers mean that real treatment opportunities are missed; cardiovascular and infection risk, however, are seen as easy to identify and act on, whereas mental illness and dementia are seen as poorly diagnosed and with limited treatment options. Prevention awareness is high for cardiovascular disease and infection, lower for cancer, and limited for dementia and mental health. Although themes overlap across diseases, the specific concerns are different and each condition has a unique profile. Quantifying the relative importance of these themes could allow their incorporation in decision-making, not only when they occur as a named disease but also in any relevant condition.


Microfluidic chip for molecular amplification of influenza A RNA in human respiratory specimens.
A rapid, low cost, accurate point-of-care (POC) device to detect influenza virus is needed for effective treatment and control of both seasonal and pandemic strains. We developed a single-use microfluidic chip that integrates solid phase extraction (SPE) and molecular amplification via a reverse transcription polymerase chain reaction (RT-PCR) to amplify influenza virus type A RNA. We demonstrated the ability of the chip to amplify influenza A RNA in human nasopharyngeal aspirate (NPA) and nasopharyngeal swab (NPS) specimens collected at two clinical sites from 2008-2010. The microfluidic test was dramatically more sensitive than two currently used rapid immunoassays and had high specificity that was essentially equivalent to the rapid assays and direct fluorescent antigen (DFA) testing. We report 96% (CI 89%,99%) sensitivity and 100% (CI 95%,100%) specificity compared to conventional (bench top) RT-PCR based on the testing of n = 146 specimens (positive predictive value = 100%(CI 94%,100%) and negative predictive value = 96%(CI 88%,98%)). These results compare well with DFA performed on samples taken during the same time period (98% (CI 91%,100%) sensitivity and 96%(CI 86%,99%) specificity compared to our gold standard testing). Rapid immunoassay tests on samples taken during the enrollment period were less reliable (49%(CI 38%,61%) sensitivity and 98%(CI 98%,100%) specificity). The microfluidic test extracted and amplified influenza A RNA directly from clinical specimens with viral loads down to 10³ copies/ml in 3 h or less. The new test represents a major improvement over viral culture in terms of turn around time, over rapid immunoassay tests in terms of sensitivity, and over bench top RT-PCR and DFA in terms of ease of use and portability.


[Evaluation of the availability, utilization, and costs of computerized tomography in the state of Morelos, Mexico].
To assess the availability, utilization, and costs of computerized tomography (CT scan) in private and public hospitals in Morelos State, Mexico. From January to April 1999, a cross-sectional study was carried out in two private and two public hospitals in Morelos, Mexico. Individual face-to-face interviews were conducted with directors, managers, radiologists, and heads of maintenance at each hospital. Statistical analysis was performed to describe the variables measuring availability, utilization, and costs of CT scans. A comprehensive assessment of CT scans was also performed. Emphasis was made on the variability of observed patterns among the participating hospitals. CT scan technology has been used by hospitals in Morelos State for over ten years; programs for preventive or corrective maintenance of these equipments are available, although at high costs. No strategies for technology assessment are available for acquisition of CT scanners nor during their period of utilization. This study did not attempt to evaluate the appropriateness of the clinical use of CT nor its untoward effects. Findings from the present study showed that: 1) a lack of mechanisms for technology assessment and management of CT scans prevented hospitals from managing CT scanning technologies efficiently; 2) technology assessment regulation is not available, even though it is necessary for the adequate selection of the best technologies, on the basis of their efficiency, effectiveness, safety, and availability.


A user-centred home monitoring and self-management system for patients with heart failure: a multicentre cohort study.
Previous generations of home monitoring systems have had limited usability. We aimed to develop and evaluate a user-centred and adaptive system for health monitoring and self-management support in patients with heart failure. Patients with heart failure were recruited from three UK centres and provided with Internet-enabled tablet computers that were wirelessly linked with sensor devices for blood pressure, heart rate, and weight monitoring. Patient observations, interviews, and concurrent analyses of the automatically collected data from their monitoring devices were used to increase the usability of the system. Of the 52 participants (median age 77 years, median follow-up 6 months [interquartile range, IQR, 3.6-9.2]), 24 (46%) had no, or very limited prior, experience with digital technologies. It took participants about 1.5 min to complete the daily monitoring tasks, and the rate of failed attempts in completing tasks was <5%. After 45 weeks of observation, participants still used the system on 4.5 days per week (confidence interval 3.2-5.7 days). Of the 46 patients who could complete the final survey, 93% considered the monitoring system as easy to use and 38% asked to keep the system for self-management support after the study was completed. We developed a user-centred home monitoring system that enabled a wide range of heart failure patients, with differing degrees of IT literacy, to monitor their health status regularly. Despite no active medical intervention, patients felt that they benefited from the reassurance and sense of connectivity that the monitoring system provided.


Gap junctions regulate the activity of AgRP neurons and diet-induced obesity in male mice.
Recent studies indicated an important role of connexins, gap junction proteins, in the regulation of metabolism. However, most of these studies focused on the glial expression of connexins, whereas the actions of connexins in neurons are still poorly investigated. Thus, the present study had the objective to investigate the possible involvement of gap junctions, and in particular connexin 43 (CX43), for the central regulation of energy homeostasis. Initially, we demonstrated that hypothalamic CX43 expression was suppressed in fasted mice. Using whole-cell patch-clamp recordings, we showed that pharmacological blockade of gap junctions induced hyperpolarization and decreased the frequency of action potentials in 50-70% of agouti-related protein (AgRP)-expressing neurons, depending on the blocker used (carbenoxolone disodium, TAT-Gap19 or Gap 26). When recordings were performed with a biocytin-filled pipette, this intercellular tracer was detected in surrounding cells. Then, an AgRP-specific CX43 knockout (AgRPΔCX43) mouse was generated. AgRPΔCX43 mice exhibited no differences in body weight, adiposity, food intake, energy expenditure and glucose homeostasis. Metabolic responses to 24 h fasting or during refeeding were also not altered in AgRPΔCX43 mice. However, AgRPΔCX43 male, but not female mice, exhibited a partial protection against high-fat diet-induced obesity, even though no significant changes in energy intake or expenditure were detected. In summary, our findings indicate that gap junctions regulate the activity of AgRP neurons, and AgRP-specific CX43 ablation is sufficient to mildly prevent diet-induced obesity specifically in males.


Self-assembly of diphenylalanine with preclick components as capping groups.
Alkyne and azide, which are commonly used in the cycloaddition reaction recognized as "click chemistry", have been used as capping groups of two engineered diphenylalanine (FF) derivatives due to their ability to form weak intermolecular interactions (i.e. dipole-π and π-π stacking). In Poc-FF-N3, alkyne and azide act as N- and C-terminal capping groups, respectively, while such positions are exchanged in N3-FF-OPrp. The self-assembly of such two synthesized peptides has been extensively studied in their "pre-click" state, considering the influence of three different factors: the peptide concentration, the polarity of the medium, and the nature of the substrate. Poc-FF-N3 assembles into microfibers that, depending on the medium and the substrate, can aggregate hierarchically in supramolecular structures with different morphologies. The most distinctive one corresponds to very stable birefringent dendritic-like microstructures, which are derived from the ordered agglomeration of microfibers. These branched supramolecular structures, which are observed under a variety of conditions, are relatively uncommon in short FF sequences. At the molecular level, Poc-FF-N3 organizes in antiparallel β-sheets stabilized by N-HO intermolecular hydrogen bonds and re-enforced by weak interactions between the azide and alkyne groups of neighbouring molecules. In contrast, N3-FF-OPrp exhibits a very poor tendency to organize into structures with a well-defined morphology. Theoretical calculations on model complexes indicate that the tendency of the latter peptide to organize into small amorphous agglomerates is due to its poor ability to form specific intermolecular interactions in comparison with Poc-FF-N3. The implications of the weak interactions induced by the alkyne and azide groups, which strengthen peptidepeptide hydrogen bonds and π-ladders due to the stacked aromatic phenyl side groups, are discussed.


Nucleotide and amino acid changes in West Nile virus strains exhibiting renal tropism in hamsters.
Recent studies have shown that West Nile virus (WNV) can induce an asymptomatic persistent infection in the kidneys of experimentally infected hamsters. The chronically infected rodents shed virus in their urine for up to 8 months, despite the disappearance of viremia and the development of high levels of neutralizing antibodies. WNV, like most members of the Japanese encephalitis virus complex (Flavivirus; Flaviviridae), is assumed to be mainly neurotropic; little is known about the genetic basis for its renal tropism. In this study, complete sequence analyses were done to compare four WNV isolates from the urines of persistently infected hamsters with the wild-type parent virus (NY 385-99). Nucleotide changes, ranging from 0.05% to 0.09%, were identified in all of the WNV isolates from urine; most of the changes were in coding regions, causing amino acid substitutions in the E, NS1, NS2B, and NS5 proteins. The genetic changes associated with renal tropism were also accompanied by a loss of virulence for hamsters and a change in plaque morphology.


Explaining stasis: microevolutionary studies in natural populations.
Microevolution, defined as a change in the genetic constitution of a population over time, is considered to be of commonplace occurrence in nature. Its ubiquity can be inferred from the observation that quantitative genetic divergence among populations usually exceeds that to be expected due to genetic drift alone, and from numerous observations and experiments consistent with local adaptation. Experimental manipulations in natural populations have provided evidence that rapid evolutionary responses may occur in the wild. However, there are remarkably few cases where direct observations of natural populations have revealed microevolutionary changes occurring, despite the frequent demonstration of additive genetic variation and strong directional selection for particular traits. Those few cases where responses congruent with expectation have been demonstrated are restricted to changes over one generation. In this article we focus on possible explanations as to why heritable traits under apparently strong directional selection often fail to show the expected evolutionary response. To date, few of these explanations for apparent stasis have been amenable to empirical testing. We describe new methods, derived from procedures developed by animal breeding scientists, which can be used to address these explanations, and illustrate the approach with examples from long-term studies of collared flycatchers (Ficedula albicollis) and red deer (Cervus elaphus). Understanding why most intensively studied natural populations do not appear to be evolving is an important challenge for evolutionary biology.


CD200-dependent and nonCD200-dependent pathways of NK cell suppression by human IVIG.
Intravenous immunoglobulin (IVIG) has been used to suppress autoimmune and inflammatory disorders by a variety of mechanisms. Recently, the CD200 tolerance-promoting signal has been found to play a role in IVIG suppression of blood natural killer (NK) cells. Further, different types of IVIG have been reported to differ in this activity, and that has been related to efficacy (and inefficacy) of treatment of women with pregnancy failure. CD200 acts by binding to CD200 receptors (C200R). The objective of this study was to determine if CD200-dependent NK suppression by IVIG involved direct binding of IVIG-associated CD200 molecules to CD200R on NK cells. Peripheral Blood Lymphocytes isolated from human blood were used as a source of NK cells to lyse Cr(51)-labelled K562 target cells in vitro in 18 and 4 h assays, and three different types of IVIG were tested for suppressive activity in the presence or absence of specific monoclonal anti-huCD200. In some experiments, CD56(+) NK cells were purified using anti-CD56 magnetic beads. Western blotting of IVIG using a specific anti-huCD200 antibody was done. Enzyme-Linked ImmunoSorbent Assays were used to measure cytokine production in NK assays. Different IVIGs showed significant differences in potency in suppressing NK cytolytic activity in vitro (mg/ml for 60% suppression, Gammagard 4.1, Gamunex 14.1, Gamimmune 20.2). For CD200-dependent suppression, Gammagard was twice as potent as Gamimmune, but equivalent to Gamunex. The presence of suppression in 4 hour assays indicated stimulation of cytokine synthesis was unlikely to explain CD200-dependent suppression. Purification of NK cells led to loss of the CD200-dependent component. Western blotting confirmed that material reactive with anti-CD200 antibody was present in Immunoglobulin G (IgG) preparations, and at a lower level in human serum that contains IgG. IVIGs are not all equipotent in suppressing NK cell cytolytic activity. CD200 associated with IVIG is an important component of suppression. CD200-dependent suppression appears to be mediated by a non-NK population that then acts on NK cells by direct contact rather than indirectly through release of immunosuppressive cytokines.


Justice from the victim's perspective.
What are the meanings of justice, as seen from the perspective of victims of violent crime? Are victims' visions of justice represented by the conventional legal system? Are they represented by restorative justice? The author engages these questions, drawing on in-depth interviews with 22 victims of violent crime. It is argued that survivors' views of justice do not fit well into either retributive or restorative models. This has implications for current efforts to use restorative models in cases of violence against women.


Age-Related Differences in Dynamic Interactions Among Default Mode, Frontoparietal Control, and Dorsal Attention Networks during Resting-State and Interference Resolution.
Resting-state fMRI (rs-fMRI) can identify large-scale brain networks, including the default mode (DMN), frontoparietal control (FPN) and dorsal attention (DAN) networks. Interactions among these networks are critical for supporting complex cognitive functions, yet the way in which they are modulated across states is not well understood. Moreover, it remains unclear whether these interactions are similarly affected in aging regardless of cognitive state. In this study, we investigated age-related differences in functional interactions among the DMN, FPN and DAN during rest and the Multi-Source Interference task (MSIT). Networks were identified using independent component analysis (ICA), and functional connectivity was measured during rest and task. We found that the FPN was more coupled with the DMN during rest and with the DAN during the MSIT. The degree of FPN-DMN connectivity was lower in older compared to younger adults, whereas no age-related differences were observed in FPN-DAN connectivity in either state. This suggests that dynamic interactions of the FPN are stable across cognitive states. The DMN and DAN were anti correlated and age-sensitive during the MSIT only, indicating variation in a task-dependent manner. Increased levels of anticorrelation from rest to task also predicted successful interference resolution. Additional analyses revealed that the degree of DMN-DAN anticorrelation during the MSIT was associated to resting cerebral blood flow (CBF) within the DMN. This suggests that reduced DMN neural activity during rest underlies an impaired ability to achieve higher levels of anticorrelation during a task. Taken together, our results suggest that only parts of age-related differences in connectivity are uncovered at rest and thus, should be studied in the functional connectome across multiple states for a more comprehensive picture.


Epidemiological Features and Outcomes of HTLV-1 Carriers Diagnosed With Cancer: A Retrospective Cohort Study in an Endemic Country.
Human T-lymphotropic virus type 1 (HTLV-1) is an endemic virus in Latin America that is directly linked to adult T-cell leukemia/lymphoma (ATL). Previous studies have suggested an oncogenic role of HTLV-1 in non-ATL neoplasms and have found higher mortality in HTLV-1 carriers without ATL. In this retrospective cohort study, HTLV-1 carriers were identified through screening at a tertiary cancer center between 2006 and 2019. We compared the overall survival (OS) outcomes of patients with ATL with those with other solid or hematologic malignancies by sex stratification. We identified 1,934 HTLV-1 carriers diagnosed with cancer. The median age at diagnosis was 62 (range 20-114) years, 76% were female, 60% had no or elementary school education, and 50% were born in the Andean highlands. The most common non-ATL neoplasm was cervical cancer (50%) among females and non-ATL non-Hodgkin lymphoma (26%) among males. With a median follow-up of 66 months, the 5-year OS of HTLV-1 carriers with non-ATL neoplasms (26%-47% for females and 22%-34% for males) was inferior to those reported in the general population. As expected, patients with ATL had a worse prognosis (5-year OS: 10% for females and 8% for males). HTLV-1 carriers with cancer were middle age and from underprivileged settings, suggesting an undetected transmission among vulnerable populations, especially females. Survival estimates of HTLV-1 carriers with non-ATL neoplasms were lower than the regional outcomes. Future research should ascertain how the biology of HTLV-1 and health care disparities affect the outcomes of HTLV-1 carriers, as well as determine the burden of HTLV-1 infection in the cancer population to recommend screening in the outpatient setting of endemic regions.


Theoretical determination of two-photon absorption in biologically relevant pterin derivatives.
Given the prevalence of fluorescence spectroscopy in biological systems, and the prevalence of pterin derivatives throughout biological systems, presented here is an assessment of the two-photon absorption spectroscopy as it applies to a range of the most commonly studied pterin derivatives. QR-CAMB3LYP//ccpVTZ calculations suggest that the use of two-photon spectroscopic methods would enable a more capable differentiation between closely related derivatives in comparison to the one-photon spectra, which show minimal qualitative deviation. Study of short tail derivatives shows that, in most cases, two-photon accessible states solely involve the π* LUMO as the particle orbital, with biopterin, neopterin, and 6-(hydroxymethyl)pterin presenting exceptional potential for targetting. Investigation of derivatives in which the tail contains an aromatic ring resulted in the observation of a series of two-photon accessible states involving charge transfer from the tail to the pterin moiety, the cross sections of which are highly dependent on the adoption of a planar geometry. The observation of these states presents a novel method for tracking the substitution of biologically important molecules such as folic acid and 5-methenyltetrahydrofolylpolyglutamate.


Lab-on-a-chip for ultrasensitive detection of carbofuran by enzymatic inhibition with replacement of enzyme using magnetic beads.
In this paper an ultrasensitive method to determine toxicity due to pesticides in a glass lab-on-a-chip by means of enzymatic inhibition of acetylcholinesterase immobilised on magnetic beads is described. The reproducible insertion of a controlled amount of enzyme-coupled magnetic beads inside the chip channel and their immobilisation in a capture region with the aid of a magnetic field has been optimised. This procedure enables the easy renewal of the biosensing material after each determination in a highly reproducible manner. Several operational parameters such as the working potential for the selective detection of thiocholine (TCh) on a platinum disc electrode, the TCh detection reproducibility and sensitivity, the electroosmotic flow driving voltage and the inhibition time were also evaluated or optimised. The detection of carbofuran (one of the most toxic carbamate pesticides) has been achieved down to the nanomolar level.


Trends in Patient Characteristics and COVID-19 In-Hospital Mortality in the United States During the COVID-19 Pandemic.
In-hospital mortality rates from COVID-19 are high but appear to be decreasing for selected locations in the United States. It is not known whether this is because of changes in the characteristics of patients being admitted. To describe changing in-hospital mortality rates over time after accounting for individual patient characteristics. This was a retrospective cohort study of 20 736 adults with a diagnosis of COVID-19 who were included in the US American Heart Association COVID-19 Cardiovascular Disease Registry and admitted to 107 acute care hospitals in 31 states from March through November 2020. A multiple mixed-effects logistic regression was then used to estimate the odds of in-hospital death adjusted for patient age, sex, body mass index, and medical history as well as vital signs, use of supplemental oxygen, presence of pulmonary infiltrates at admission, and hospital site. In-hospital death adjusted for exposures for 4 periods in 2020. The registry included 20 736 patients hospitalized with COVID-19 from March through November 2020 (9524 women [45.9%]; mean [SD] age, 61.2 [17.9] years); 3271 patients (15.8%) died in the hospital. Mortality rates were 19.1% in March and April, 11.9% in May and June, 11.0% in July and August, and 10.8% in September through November. Compared with March and April, the adjusted odds ratios for in-hospital death were significantly lower in May and June (odds ratio, 0.66; 95% CI, 0.58-0.76; P < .001), July and August (odds ratio, 0.58; 95% CI, 0.49-0.69; P < .001), and September through November (odds ratio, 0.59; 95% CI, 0.47-0.73). In this cohort study, high rates of in-hospital COVID-19 mortality among registry patients in March and April 2020 decreased by more than one-third by June and remained near that rate through November. This difference in mortality rates between the months of March and April and later months persisted even after adjusting for age, sex, medical history, and COVID-19 disease severity and did not appear to be associated with changes in the characteristics of patients being admitted.


Abnormal IL-4 gene expression by atopic dermatitis T lymphocytes is reflected in altered nuclear protein interactions with IL-4 transcriptional regulatory element.
Among the atopic disease, atopic dermatitis is characterized by the highest levels of serum IgE and by increased peripheral blood T-cell interleukin-4 (IL-4) production. IL-4 promotes IgE synthesis by B cells and stimulates the growth of IL-4-producing T cells and may contribute to the pathogenesis of this disease. In this study, in situ hybridization established that atopic dermatitis patients have a higher frequency of IL-4-producing peripheral blood T cell when compared to normal subjects. These in vivo-derived T cells were used to examine the signaling requirements of IL-4 production and the nuclear factors that associate with a critical IL-4 transcriptional regulatory element between -88 and -60 relative to the IL-4 transcription initiation site, the activation responsive element. We demonstrate that, as in T-cell lines, proteins belonging to the NF-AT and AP-1 family of transcription factors are present in stimulated cell extracts and specifically associate with the activation responsive element. Dysregulated IL-4 production is reflected in the nuclear proteins that associated this element. Using gel shift assays, we found that 12 of 12 nuclear extracts from stimulated atopic T cells formed the activation-dependent protein-DNA complex, compared to only 2 of 12 normal T-cell extracts. Activation complex formation correlated with the relative level of IL-4 mRNA and protein produced in stimulated T cells, suggesting that abnormal IL-4 gene expression in atopic disease may be linked to alterations in nuclear protein interactions with these promoter elements.