Thermal injury in the elderly: when is comfort care the right choice?
The factors contributing to a higher mortality rate in elderly thermal injury victims are not well delineated. The purpose of this study is to determine the impact of the initial injury, medical comorbidities, and burn size on patient outcome and to determine a level of injury in this population when comfort care is an appropriate first choice. Individual medical records of patients over 65 years of age admitted to our burn center over a 10-year interval were reviewed for patient demographics, mechanism of injury, total body surface area (TBSA) burned, medical comorbidities, use of Swan-Ganz catheters, evidence of inhalation injury, level of support, and patient outcome. The mechanisms of thermal injury were flame (68%), scald (21%) and electrical or chemical contact (11%). Twenty-six preventable bathing, cooking, and smoking-related injuries were seen (33%). The average TBSA was 25 per cent. Average length of stay varied depending on outcome. The overall mortality rate for this group was 45 per cent. Patients older than 80 years with 40 per cent or greater TBSA burned had a 100 per cent mortality rate despite aggressive treatment. Burn wound size correlated better with probability of poor outcome than age. Thermal injuries in the elderly are becoming more important with the aging of our population. Underlying medical problems--specifically chronic obstructive pulmonary disease--do play a role in increased patient morbidity and mortality. This study shows that age greater than 80 years in combination with burns greater than 40 per cent TBSA are uniformly fatal despite aggressive therapy. We believe that delaying the start of comfort-only measures in this situation only prolongs the pain and suffering for the patient, the family, and the physician.


In vitro denervation of the portal vein and caudal artery of the rat.
The effects of 6-hydroxydopamine (6-OHDA) on the isolated portal vein and caudal artery of the rat were studied to investigate the possibility of producing in vitro adrenergic denervation of these blood vessels. Loss of nerve function was determined by field stimulation of the nerves, using short pulses, and by 3H-norepinephrine (NE) uptake. Addition of 6-OHDA produced contractions of both veins and arteries. Two hours after treatment with 6-OHDA, the contractile responses of the caudal arteries and portal veins to field stimulation were reduced to undetectable levels. At this point, the vessels were unable to take up 3H-NE and incubation of the preparations with l-NE failed to restore the contractile responses to nerve stimulation. Prejunctional supersensitivity to l-NE was observed in the portal vein after 6-OHDA but hot in helically cut strips of caudal artery. Prejunctional supersensitivity of the caudal artery to NE was seen, however, if the vessel geometry was kept intact, suggesting that the uptake mechanism for catecholamines only plays a major role in the termination of action of exgenous NE when norepinephrine is applied through the nerve plexus. We conclude that in vitro treatment with 6-OHDA rapidly produces functional adrenergic denervation of both portal vein and caudal artery of the rat and provides an accurate assessment of the importance of the neuronal uptake mechanism to NE sensitivity.


Phosphorylation at S87 is enhanced in synucleinopathies, inhibits alpha-synuclein oligomerization, and influences synuclein-membrane interactions.
Increasing evidence suggests that phosphorylation may play an important role in the oligomerization, fibrillogenesis, Lewy body (LB) formation, and neurotoxicity of alpha-synuclein (alpha-syn) in Parkinson disease. Herein we demonstrate that alpha-syn is phosphorylated at S87 in vivo and within LBs. The levels of S87-P are increased in brains of transgenic (TG) models of synucleinopathies and human brains from Alzheimer disease (AD), LB disease (LBD), and multiple system atrophy (MSA) patients. Using antibodies against phosphorylated alpha-syn (S129-P and S87-P), a significant amount of immunoreactivity was detected in the membrane in the LBD, MSA, and AD cases but not in normal controls. In brain homogenates from diseased human brains and TG animals, the majority of S87-P alpha-syn was detected in the membrane fractions. A battery of biophysical methods were used to dissect the effect of S87 phosphorylation on the structure, aggregation, and membrane-binding properties of monomeric alpha-syn. These studies demonstrated that phosphorylation at S87 expands the structure of alpha-syn, increases its conformational flexibility, and blocks its fibrillization in vitro. Furthermore, phosphorylation at S87, but not S129, results in significant reduction of alpha-syn binding to membranes. Together, our findings provide novel mechanistic insight into the role of phosphorylation at S87 and S129 in the pathogenesis of synucleinopathies and potential roles of phosphorylation in alpha-syn normal biology.


Sodium-activated potassium current in sensory neurons: a comparison of cell-attached and cell-free single-channel activities.
Single-channel currents from Na(+)-dependent K+ channels (KNa) were recorded from cell-attached and inside-out membrane patches of cultured avian trigeminal ganglion neurons by means of the patch-clamp technique. Single-channel properties, such as the high elementary conductance and the occurrence of sub-conductance levels, were unchanged after the patches had been excised from the cells, indicating that they are not under the control of soluble cytoplasmic factors. In cell-attached recordings at the cell resting potential the degree of KNa activity, measured as the probability of the channel being open, Po, was low in most cases (around 0.01) and similar to that observed in the inside-out configuration when the bath solution contained concentrations of Na+ around 30 mM and of K+ close to the physiological intracellular levels. However, in some cell-attached patches Po was high (around 0.2) and comparable to the values measured in cell-free recordings with high Na+ concentrations in the bath (100 mM). The excision of a high-activity patch in the presence of 30 mM Na+ resulted in a fall of Po in about 20 s, which is consistent with the wash-out of a soluble cytoplasmic molecule. After the excision, all KNa displayed a similar Na+ sensitivity, irrespective of the degree of activation observed in the cell-attached mode. In inside-out patches the Po values observed in the presence of either low or high concentrations of Na+ in bath solutions were not modified by internal Ca2+ (0.8-8.5 microM).(ABSTRACT TRUNCATED AT 250 WORDS)


Age-related impairment of declarative memory: linking memorization of temporal associations to GluN2B redistribution in dorsal CA1.
GluN2B subunits of NMDA receptors have been proposed as a target for treating age-related memory decline. They are indeed considered as crucial for hippocampal synaptic plasticity and hippocampus-dependent memory formation, which are both altered in aging. Because a synaptic enrichment in GluN2B is associated with hippocampal LTP in vitro, a similar mechanism is expected to occur during memory formation. We show instead that a reduction of GluN2B synaptic localization induced by a single-session learning in dorsal CA1 apical dendrites is predictive of efficient memorization of a temporal association. Furthermore, synaptic accumulation of GluN2B, rather than insufficient synaptic localization of these subunits, is causally involved in the age-related impairment of memory. These challenging data identify extra-synaptic redistribution of GluN2B-containing NMDAR induced by learning as a molecular signature of memory formation and indicate that modulating GluN2B synaptic localization might represent a useful therapeutic strategy in cognitive aging.


Possible long-lasting inhibition of converting enzyme by enalapril in human cerebrospinal fluid.
1. Converting enzyme and neutral endopeptidase activities were both measured every 3 h by a fluorimetric method in plasma and cerebrospinal fluid of patients diagnosed as migraineurs with aura after lumbar puncture, which was performed 9 h after an acute oral dose of enalapril or placebo. 2. A reduced converting enzyme activity, as compared with placebo, was observed in patients who were given enalapril. On the other hand, neutral endopeptidase activity detected after enalapril did not differ from that measured after placebo. 3. The results seem to indicate that enalapril penetrates the blood-brain barrier in sufficient amounts to reduce converting enzyme activity. Moreover, neutral endopeptidase was not affected by enalapril. Therefore, those clinical effects of the drug which have been attributed to the involvement of central opioid mechanisms may depend on the inhibition of brain converting enzyme but not the inhibition of brain neutral endopeptidase.


Investigation of the porous structure of cellulosic substrates through confocal laser scanning microscopy.
At the most fundamental level, saccharification occurs when cell wall degrading enzymes (CWDEs) diffuse, bind to and react on readily accessible cellulose fibrils. Thus, the study of the diffusive behavior of solutes into and out of cellulosic substrates is important for understanding how biomass pore size distribution affects enzyme transport, binding, and catalysis. In this study, fluorescently labeled dextrans with molecular weights of 20, 70, and 150 kDa were used as probes to assess their diffusion into the porous structure of filter paper. Fluorescence microscopy with high numerical aperture objectives was used to generate high temporal and spatial resolution datasets of probe concentrations versus time. In addition, two diffusion models, including a simple transient diffusion and a pore grouping diffusion models, were developed. These models and the experimental datasets were used to investigate solute diffusion in macro- and micro-pores. Nonlinear least squares fitting of the datasets to the simple transient model yielded diffusion coefficient estimates that were inadequate for describing the initial fast diffusion and the later slow diffusion rates observed; on the other hand, nonlinear least squares fitting of the datasets to the pore grouping diffusion model yielded estimations of the micro-pore diffusion coefficient that described the inherently porous structure of plant-derived cellulose. In addition, modeling results show that on average 75% of the accessible pore volume is available for fast diffusion without any significant pore hindrance. The method developed can be applied to study the porous structure of plant-derived biomass and help assess the diffusion process for enzymes with known sizes.


Environmental effects of ozone depletion and its interactions with climate change: progress report, 2009.
The parties to the Montreal Protocol are informed by three panels of experts. One of these is the Environmental Effects Assessment Panel (EEAP), which deals with UV radiation and its effects on human health, animals, plants, biogeochemistry, air quality and materials. Since 2000, the analyses and interpretation of these effects have included interactions between UV radiation and global climate change. When considering the effects of climate change, it has become clear that processes resulting in changes in stratospheric ozone are more complex than believed previously. As a result of this, human health and environmental problems will likely be longer-lasting and more regionally variable. Like the other panels, the EEAP produces a detailed report every four years; the most recent was that for 2006 (Photochem. Photobiol. Sci., 2007, 6, 201-332). In the years in between, the EEAP produces a less detailed and shorter progress report, as is the case for this present one for 2009. A full quadrennial report will follow for 2010.


Mistreatment of women in public health facilities of Ethiopia.
Recent evidence suggests that mistreatment of women during childbirth is a global challenge facing health care systems. This study seeks to explore the prevalence of mistreatment of women in public health facilities of Ethiopia, and identify associated factors. A two-stage cross sectional sampling design was used to select institutions and women. The study was conducted in hospitals and health centers across four Ethiopian regions. Quantitative data were collected from postpartum women. Mistreatment was measured using four domains: (1) physical abuse, (2) verbal abuse, (3) failure to meet professional standards of care, and (4) poor rapport between women and providers. Percentages of mistreatment and odds ratios for the association between its presence and institutional and socio demographic characteristics of women were calculated using bivariate and multivariable logistic regression modeling. A total of 379 women were interviewed, of whom 281 (74%) reported any mistreatment. Physical and verbal abuse were reported by 7 (2%) and 31 (8%) women interviewed respectively. Failure to meet professional standards of care and poor rapport between women and providers were reported by 111 (29%) and 274 (72%) women interviewed respectively. Multivariable logistic regression analysis revealed that the odds of reporting mistreatment were higher among women with four or more previous births (aOR = 3.36 95%CI 1.22,9.23, p = 0.019) compared to women with no previous childbirth, Muslim women (aOR = 3.30 95%CI 1.4,7.77, p = 0.006) and women interviewed in facilities with less than 17 births per MNH staff in a month (aOR = 3.63 95%CI 1.9,6.93, p < 0.001). However, the odds of reporting mistreatment were lower among women aged 35 and older (aOR = 0.22 95%CI 0.06, 0.73, p = 0.014) and among women interviewed between 8 and 42 days after childbirth (aOR = 0.37 95%CI 0.15, 0.9, p = 0.028). Mistreatment during childbirth in Ethiopia is commonly reported. Health workers need to consider provision of individualized care for women and monitor their experiences in order to adjust quality of their services.


Normal limits of the electrocardiogram in Chinese subjects.
Accurate normal limits of the electrocardiogram (ECG) are the basis on which diagnostic criteria are developed. The ECG, however, is subject to age- and sex-variations and may also be racially determined. Studies into normal ECG limits for the Chinese, comprising one fifth of the world population, are few and have their limitations. We have undertaken to establish normal limits of the ECG from a large sample of healthy Chinese subjects. Standard simultaneous 12-lead ECGs from 5360 apparently healthy Chinese subjects (3614 men and 1746 women, ages ranging from 18 to 84 years) were collected with a modern digital recorder and processed with a well-validated ECG computer program. The medians, lower limits (2nd percentile) and upper limits (98th percentile) of various ECG measurements were calculated and age and sex differences examined. Significant age trends were present in, for example, P-wave duration, QTc interval, and frontal QRS axis, with concomitant changes of R amplitudes in the extremity leads. Sex differences existed for heart rate, interval durations, the Sokolow and Cornell indices, and QRS and ST-T amplitudes in different leads. Notably, left-precordial R-wave amplitudes in women increased with age; the Sokolow index showed a clearer age trend for men than for women, the reverse being true for the Cornell index. Some of these findings are at odds with established diagnostic ECG criteria. Normal ECG limits of Chinese subjects show marked age and sex differences. This merits the definition and use of age- and sex-specific ECG criteria for a Chinese population.


Sensing of minute airflow motions near walls using pappus-type nature-inspired sensors.
This work describes the development and use of pappus-like structures as sensitive sensors to detect minute air-flow motions. We made such sensors from pappi taken from nature-grown seed, whose filiform hairs' length-scale is suitable for the study of large-scale turbulent convection flows. The stem with the pappus on top is fixated on an elastic membrane on the wall and tilts under wind-load proportional to the velocity magnitude in direction of the wind, similar as the biological sensory hairs found in spiders, however herein the sensory hair has multiple filiform protrusions at the tip. As the sensor response is proportional to the drag on the tip and a low mass ensures a larger bandwidth, lightweight pappus structures similar as those found in nature with documented large drag are useful to improve the response of artificial sensors. The pappus of a Dandelion represents such a structure which has evolved to maximize wind-driven dispersion, therefore it is used herein as the head of our sensor. Because of its multiple hairs arranged radially around the stem it generates uniform drag for all wind directions. While still being permeable to the flow, the hundreds of individual hairs on the tip of the sensor head maximize the drag and minimize influence of pressure gradients or shear-induced lift forces on the sensor response as they occur in non-permeable protrusions. In addition, the flow disturbance by the sensor itself is limited. The optical recording of the head-motion allows continuously remote-distance monitoring of the flow fluctuations in direction and magnitude. Application is shown for the measurement of a reference flow under isothermal conditions to detect the early occurrence of instabilities.


Recursive model for dose-time responses in pharmacological studies.
Clinical studies often track dose-response curves of subjects over time. One can easily model the dose-response curve at each time point with Hill equation, but such a model fails to capture the temporal evolution of the curves. On the other hand, one can use Gompertz equation to model the temporal behaviors at each dose without capturing the evolution of time curves across dosage. In this article, we propose a parametric model for dose-time responses that follows Gompertz law in time and Hill equation across dose approximately. We derive a recursion relation for dose-response curves over time capturing the temporal evolution and then specify a regression model connecting the parameters controlling the dose-time responses with individual level proteomic data. The resultant joint model allows us to predict the dose-response curves over time for new individuals. We have compared the efficacy of our proposed Recursive Hybrid model with individual dose-response predictive models at desired time points. We note that our proposed model exhibits a superior performance compared to the individual ones for both synthetic data and actual pharmacological data. For the desired dose-time varying genetic characterization and drug response values, we have used the HMS-LINCS database and demonstrated the effectiveness of our model for all available anticancer compounds.


Dynamics of linear polarization conversion in uniaxial crystals.
We report on the experimental and theoretical investigation of polarization conversion of linearly polarized Gaussian beam propagating in perpendicularly cut homogeneous uniaxial crystals. We derive analytical expressions, in good agreement with experimental data, for power transfer between components at normal incidence accompanied by the generation of a topological quadrupole. We extend the results to the oblique incidence case and confirm experimentally the optimal parameters for generation of a single charge on-axis optical vortex, including spectrally resolved measurements for the white-light beams.


Resolution of metabolic syndrome after following a gluten free diet in an adult woman diagnosed with celiac disease.
Adult celiac disease (CD) presents with very diverse symptoms that are clearly different from those typically seen in pediatric patients, including ferropenic anemia, dyspepsia, endocrine alterations and elevated transaminase concentration. We present the case of a 51-year-old overweight woman with altered basal blood glucose, hypercholesterolemia, hypertriglyceridemia and persisting elevated transaminase levels, who showed all the symptoms for a diagnosis of metabolic syndrome. Because she presented iron deficiency anemia, she was referred to the gastroenterology department and subsequently diagnosed with celiac disease after duodenal biopsies and detection of a compatible HLA haplotype. Gluten-free diet (GFD) was prescribed and after 6 mo the patient showed resolution of laboratory abnormalities (including recovering anemia and iron reserves, normalization of altered lipid and liver function parameters and decrease of glucose blood levels). No changes in weight or waist circumference were observed and no significant changes in diet were documented apart from the GFD. The present case study is the first reported description of an association between CD and metabolic syndrome, and invites investigation of the metabolic changes induced by gluten in celiac patients.


UltraRapid communications : vascular superoxide production by NAD(P)H OxidaseAssociation with endothelial dysfunction and clinical risk factors
Superoxide anion plays important roles in vascular disease states. Increased superoxide production contributes to reduced nitric oxide (NO) bioactivity and endothelial dysfunction in experimental models of vascular disease. We measured superoxide production by NAD(P)H oxidase in human blood vessels and examined the relationships between NAD(P)H oxidase activity, NO-mediated endothelial function, and clinical risk factors for atherosclerosis. Endothelium-dependent vasorelaxations and direct measurements of vascular superoxide production were determined in human saphenous veins obtained from 133 patients with coronary artery disease and identified risk factors. The predominant source of vascular superoxide production was an NAD(P)H-dependent oxidase. Increased vascular NAD(P)H oxidase activity was associated with reduced NO-mediated vasorelaxation. Furthermore, reduced endothelial vasorelaxations and increased vascular NAD(P)H oxidase activity were both associated with increased clinical risk factors for atherosclerosis. Diabetes and hypercholesterolemia were independently associated with increased NADH-dependent superoxide production. The association of increased vascular NAD(P)H oxidase activity with endothelial dysfunction and with clinical risk factors suggests an important role for NAD(P)H oxidase-mediated superoxide production in human atherosclerosis. The full text of this article is available at http://www.circresaha.org. Key Words:atherosclerosis endothelium superoxide nitric oxide diabetes Two Distinct Congenital Arrhythmias Evoked by a Multidysfunctional Na(+) Channel Marieke W. Veldkamp, Prakash C. Viswanathan, Connie Bezzina, Antonius Baartscheer, Arthur A.M. Wilde, Jeffrey R. Balser Abstract-The congenital long-QT syndrome (LQT3) and the Brugada syndrome are distinct, life-threatening rhythm disorders linked to autosomal dominant mutations in SCN5A, the gene encoding the human cardiac Na(+) channel. It is believed that these two syndromes result from opposite molecular effects: LQT3 mutations induce a gain of function, whereas Brugada syndrome mutations reduce Na(+) channel function. Paradoxically, an inherited C-terminal SCN5A mutation causes affected individuals to manifest electrocardiographic features of both syndromes: QT-interval prolongation (LQT3) at slow heart rates and distinctive ST-segment elevations (Brugada syndrome) with exercise. In the present study, we show that the insertion of the amino acid 1795insD has opposite effects on two distinct kinetic components of Na(+) channel gating (fast and slow inactivation) that render unique, simultaneous effects on cardiac excitability. The mutation disrupts fast inactivation, causing sustained Na(+) current throughout the action potential plateau and prolonging cardiac repolarization at slow heart rates. At the same time, 1795insD augments slow inactivation, delaying recovery of Na(+) channel availability between stimuli and reducing the Na(+) current at rapid heart rates. Our findings reveal a novel molecular mechanism for the Brugada syndrome and identify a new dual mechanism whereby single SCN5A mutations may evoke multiple cardiac arrhythmia syndromes by influencing diverse components of Na(+) channel gating function. The full text of this article is available at http://www.circresaha.org. Key Words: Na(+) channel inactivation long-QT syndrome Brugada syndrome


Disgust and Huntington's disease.
The disproportionate impairment for the recognition of facial expressions of disgust in patients with Huntington's disease (HD) forms a double dissociation with the impaired recognition of fear that has been reported in amygdala patients. The dissociation has generated discussion regarding the potential existence of neural substrates dedicated to the recognition of facial signals of specific emotions. The aim of this study was to establish whether the impairment for disgust in HD was restricted solely to the domain of facial perception, or whether HD patients also demonstrate impairment in other kinds of disgust. Fourteen HD patients and fourteen age and education matched healthy controls participated in seven disparate emotion processing tasks. (1) A measure of knowledge for the situational determinants of distinct emotions; (2) recognition of emotion expressed in nonverbal vocalisations; (3) recognition of the emotional content of explicit lexical stimuli; (4) recognition of emotional content in pictures of emotion scenes; (5) a disgust experience questionnaire; (6) a measure of olfactory hedonic responsiveness; (7) a measure of gustatory perception. While verbal aspects of disgust processing were preserved, parallel impairments were revealed for olfactory disgust, vocal disgust expressions, the classification of disgusting pictures, and declarative knowledge of disgust elicitors. The finding of impaired perception of disgust signalled through different input domains suggests that the inability to recognise the facial expression in this population reflects a fundamental problem with disgust processing.


The effect of grazing exclusion over time on structure, biodiversity, and regeneration of high nature value farmland ecosystems in Europe.
Climate change and increasing socio-economic pressure is placing many ecosystems of high ecological and economic value at risk. This is particularly urgent in dryland ecosystems, such as the montado, a multifunctional savannah-like system heavily modeled by grazing. There is still an ongoing debate about the trade-offs between livestock grazing and the potential for ecosystem regeneration. While it is consensual that overgrazing hinders the development of the shrubs and trees in this system, the effects of undergrazing or grazing exclusion are unclear. This study provides the unique opportunity to study the impact of grazing on compositional and structural biodiversity by examining the ecological chronosequence in a long-term ecological research site, located in Portugal, where grazing exclusion was controlled for over 15years. As the threat of intensification persists, even in areas where climate shifts are evident, there is a critical need to understand if and how the montado might recover by removing grazing pressure. We evaluate succession on structural and compositional diversity after grazing pressure is removed from the landscape at 5, 10, and 15years post-cattle exclusion and contrast it with currently grazed plots. A LiDAR-derived structural diversity index (LHDI), a surrogate of ecosystem structure and function first developed for the pine-grassland woodland systems, is used to quantify the impact of grazing exclusion on structure and natural regeneration. The distribution of the vegetation, particularly those of the herbaceous and shrub strata (>10≤150cm), presents statistically significant changes. The LHDI closely mimics the compositional biodiversity of the shrubs, with an increase in diversity with increased years without grazing. Under present climate conditions, both shrub regeneration and the establishment of tree saplings were strongly promoted by grazing exclusion, which has important management implications for the long-term sustainability of montado systems.


Evaluation of a radiographic caudolateral curvilinear osteophyte on the femoral neck and its relationship to degenerative joint disease and distraction index in dogs.
To determine prevalence of a radiographic caudolateral curvilinear osteophyte (CCO) on the femoral neck in various breeds and age groups of dogs and to evaluate its contemporaneous relationship with degenerative joint disease (DJD) and distraction index (DI). Cross-sectional prevalence study. 25,968 dogs, including 3,729 German Shepherd Dogs, 4,545 Golden Retrievers, 6,277 Labrador Retrievers, and 1,191 Rottweilers. Data from the University of Pennsylvania Hip Improvement Program database were analyzed, including ventrodorsal hip-extended, compression, and distraction radiographs. The CCO and radiographic signs of DJD were considered independent events and were interpreted as either present or absent. Statistical methods were used to evaluate the CCO as a possible risk factor for DJD and assess its association with DI, as measured by use of distraction radiography. When all breeds were pooled, DJD was detected in 8.6% of dogs, and the CCO was detected in 21.6% of dogs. Among dogs with a CCO, 25.1% had radiographic evidence of DJD. Among dogs without a CCO, only 4% had DJD. Dogs with a CCO were 7.9 times as likely to have DJD as were those without a CCO. Additionally, DI, weight, and age were significant risk factors for the CCO. Results confirm the contemporaneous association between the CCO and DJD and that passive hip laxity, as measured by use of the DI, is associated with both the CCO and DJD.


p38 MAP kinase activation mediates gamma-globin gene induction in erythroid progenitors.
Our goal was to determine the role of p38 mitogen-activated protein kinase (MAPK) signaling in fetal hemoglobin (HbF) induction. Two histone deacetylase inhibitors (HDAIs), sodium butyrate (NB), and trichostatin (TSA) and hemin were analyzed. In addition, the effect of direct activation of p38 MAPK on gamma-globin gene activity was studied. Primary erythroid progenitors derived from peripheral blood mononuclear cell and K562 erythroleukemia cells were analyzed. Cells were grown in NB, TSA, hemin, or anisomycin either alone or in the presence of the p38 MAPK inhibitor SB203580. The effects of the various treatments on gamma-globin RNA, HbF, and phosphorylated p38 MAPK levels were measured by RNase protection assay, alkaline denaturation, and Western blot analysis, respectively. A K562 stable line overexpressing constitutively active p38 MAPK was established using MAPK kinase kinase 3 (MKK3) and MKK6, the immediate upstream activators of p38. The direct effect of p38 MAPK overexpression on gamma-globin mRNA synthesis was analyzed. NB and TSA activated p38 MAPK and increased gamma-globin mRNA levels in K562 cells and primary erythroid progenitors. Pretreatment with SB203580 blocked p38 MAPK and gamma-globin gene activation. In contrast, no change in p38 activity was observed with hemin inductions. Direct activation of p38 by anisomycin or constitutive overexpression also increased gamma-globin mRNA in the absence of HbF inducers in wild-type K562 cells and in the MKK stable lines. This study supports a novel role for p38 MAPK in gamma-globin regulation in human erythroid progenitors.


Nascent polypeptide chains exit the ribosome in the same relative position in both eucaryotes and procaryotes.
We located the polypeptide nascent chain as it leaves cytoplasmic ribosomes from the plant Lemna gibba by immune electron microscopy using antibodies against the small subunit of the enzyme ribulose-1,5-bisphosphate carboxylase. Similar studies with Escherichia coli ribosomes, using antibodies directed against the enzyme beta-galactosidase, show that the polypeptide nascent chain emerges in the same relative position in plants and bacteria. The eucaryotic ribosomal exit site is on the large subunit, approximately 75 A from the interface between subunits and nearly 160 A from the central protuberance, the presumed site for peptidyl transfer. This is the first functional site on both the eucaryotic and procaryotic ribosomes to be determined.


Subsurface chemical nanoidentification by nano-FTIR spectroscopy.
Nano-FTIR spectroscopy based on Fourier transform infrared near-field spectroscopy allows for label-free chemical nanocharacterization of organic and inorganic composite surfaces. The potential capability for subsurface material analysis, however, is largely unexplored terrain. Here, we demonstrate nano-FTIR spectroscopy of subsurface organic layers, revealing that nano-FTIR spectra from thin surface layers differ from that of subsurface layers of the same organic material. Further, we study the correlation of various nano-FTIR peak characteristics and establish a simple and robust method for distinguishing surface from subsurface layers without the need of theoretical modeling or simulations (provided that chemically induced spectral modifications are not present). Our experimental findings are confirmed and explained by a semi-analytical model for calculating nano-FTIR spectra of multilayered organic samples. Our results are critically important for the interpretation of nano-FTIR spectra of multilayer samples, particularly to avoid that geometry-induced spectral peak shifts are explained by chemical effects.


Preparation of novel anionic polymeric ionic liquid materials and their potential application to protein adsorption.
A series of anionic polymeric ionic liquid (PIL) materials with high adsorption capacities toward proteins were developed using a 2-acrylamido-2-methylpropane sulfonic acid imidazole derivative ([AMPS][RIm]) (R: -H, -C6H5, or -(CH2)3CH3) as the functional monomer. An anionic polymeric ionic liquid monomer (ILM) containing imidazole cations modified with alkyl derivatives (butyl or phenyl groups) and sulfonic group-functionalized anions was chosen as the only monomer to prepare materials for protein adsorption. The resultant anionic PIL materials were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. The adsorption behaviors of the obtained anionic PIL materials toward proteins was evaluated. The results indicated that the anionic PILs material showed superior adsorption for different proteins at different pH values: ovotransferrin (OVT, 861.5 mg g-1) and bovine serum albumin (BSA, 842.2 mg g-1) at pH 4.0; bovine hemoglobin (BHb, 983.4 mg g-1) and cytochrome c (Cytc, 882.9 mg g-1) at pH 7.0; and casein (605.2 mg g-1) at pH 8.0. The feasibility of the resultant material for separating BHb from real bovine blood has also been demonstrated. The convenient preparation of anionic PIL materials and ultra-high adsorption capacity toward proteins predicted its attractive and potential broad applications in proteomics, medical diagnosis, and sensors.


Living is information processing: from molecules to global systems.
We extend the concept that life is an informational phenomenon, at every level of organisation, from molecules to the global ecological system. According to this thesis: (a) living is information processing, in which memory is maintained by both molecular states and ecological states as well as the more obvious nucleic acid coding; (b) this information processing has one overall function-to perpetuate itself; and (c) the processing method is filtration (cognition) of, and synthesis of, information at lower levels to appear at higher levels in complex systems (emergence). We show how information patterns, are united by the creation of mutual context, generating persistent consequences, to result in 'functional information'. This constructive process forms arbitrarily large complexes of information, the combined effects of which include the functions of life. Molecules and simple organisms have already been measured in terms of functional information content; we show how quantification may be extended to each level of organisation up to the ecological. In terms of a computer analogy, life is both the data and the program and its biochemical structure is the way the information is embodied. This idea supports the seamless integration of life at all scales with the physical universe. The innovation reported here is essentially to integrate these ideas, basing information on the 'general definition' of information, rather than simply the statistics of information, thereby explaining how functional information operates throughout life.


Metoprolol-induced reduction in postinfarction mortality: pooled results from five double-blind randomized trials.
Several postinfarction trials have evaluated the effect of secondary prophylaxis with different beta-blockers. Although so called meta-analysis of the results from all the trials have shown a beneficial effect of postinfarction beta-blockade, many of the individual studies have shown inconclusive results, mainly due to low statistical power. In order to obtain an evaluation of the merits of postinfarction therapy with metoprolol, data from the five available studies with metoprolol have been pooled into one database. In the total material 5474 patients (4353 men, 1121 women) have been studied during double-blind therapy with metoprolol 100 mg twice daily or matching placebo. The follow-up ranges from 3 months to 3 years. In total 4732 patient years of observation have been obtained. In total there were 223 deaths in the placebo-treated patients as compared to 188 deaths in the metoprolol-treated patients (P = 0.036), which corresponds to mortality rates of 97.0 and 78.3 per 1000 patient years, respectively. The mortality reduction was found both in men and women. As has been reported from individual postinfarction beta-blocker trials, the pooled results showed a marked reduction in sudden deaths (104 in the placebo group, 62 in the metoprolol group, P = 0.002). In a Cox regression model the influence of sex, age and smoking habits on the effect of metoprolol was evaluated. None of these factors influenced the metoprolol effect significantly. It is concluded that metoprolol therapy after acute myocardial infarction reduces the total number of deaths, and especially sudden cardiac deaths. The mortality reduction was independent of gender, age and smoking habits. Available data support a continuous beneficial effect.


Whole transcriptome microarrays identify long non-coding RNAs associated with cardiac hypertrophy.
Long non-coding RNAs (lncRNAs) have recently emerged as a novel group of non-coding RNAs able to regulate gene expression. While their role in cardiac disease is only starting to be understood, their involvement in cardiac hypertrophy is poorly known. We studied the association between lncRNAs and left ventricular hypertrophy using whole transcriptome microarrays. Wild-type mice and mice overexpressing the adenosine A2A receptor were subjected to transverse aortic constriction (TAC) to induce left ventricular hypertrophy. Expression profiles of lncRNAs in the heart were characterized using genome-wide microarrays. An analytical pipeline was specifically developed to extract lncRNA data from microarrays. We identified 2 lncRNAs up-regulated and 3 lncRNAs down-regulated in the hearts of A2A-receptor overexpressing-mice subjected to TAC compared to wild-type mice. Differential expression of these 2 lncRNAs was validated by quantitative PCR. Complete microarray dataset is available at Gene Expression Omnibus (GEO) database (http://www.ncbi.nlm.nih.gov/geo/) under the accession number GSE45423. Here, we describe in details the experimental design, microarray performance and analysis.


Use of a new global indicator for vaccine safety surveillance and trends in adverse events following immunization reporting 2000-2015.
Reporting of adverse events following immunization (AEFI) is a key component for functional vaccine safety monitoring system. The aim of our study is to document trends in the AEFI reporting ratio globally and across the six World Health Organization (WHO) regions. We describe the number of AEFI reports communicated each year through the World Health Organization/United Nations Children's Fund Joint Reporting Form on Immunization from 2000 to 2015. The AEFI reporting ratios (annual AEFI reports per 100,000 surviving infants) were calculated to identify WHO countries (n = 191 in 2000 and n = 194 by 2015) that met a minimal reporting ratio of 10, a target set by the Global Vaccine Action Plan for vaccine safety monitoring as a proxy measure for a functional AEFI reporting system. The number of countries reporting any AEFI fluctuated over time but with progress from 32 (17%) in 2000 to 124 (64%) in 2015. In 2015, the global average AEFI reporting ratio was 549 AEFI reports per 100,000 surviving infants. The number of countries with AEFI reporting ratios greater than 10 increased from 8 (4%) in 2000 to 81 (42%) in 2015. In 2015, 60% of countries in the WHO Region of the Americas reported at least 10 AEFI per 100,000 surviving infants, followed by 55% in European Region, 43% in Eastern Mediterranean Region, 33% in Western Pacific Region, 27% in South-East Asia Region and 21% in African Region. Overall, AEFI reporting has increased over the past sixteen years worldwide, but requires strengthening in a majority of low- and middle- income countries. The AEFI reporting ratio is useful for benchmarking and following trends over time; but does not provide information on the quality of the reporting system and does not guarantee capacity to detect and manage a vaccine safety problem at a national level. Additional efforts are required to ensure and improve data quality, AEFI reporting and surveillance of immunization safety in every country.


Embryos, the principle of proportionality, and the shaky ground of moral respect.
The debate concerning the moral permissibility of using human embryos in human embryonic stem cell (hESC) research has long centred on the question of the embryo's supposed right to life. However, in focussing only on this question, many opponents to hESC research have escaped rigorous scrutiny by making vague and unfounded appeals to the concept of moral respect in order to justify their opposition to certain hESC practices. In this paper, I offer a critical analysis of the concept of moral respect, and its use to support the intuitively appealing principle of proportionality in hESC research. I argue that if proponents of this principle are to justify its adoption by appealing to the concept of moral respect, they must explain two things concerning the nature of the moral respect owed to embryos. First, they must explain which particular aspect of the embryo is morally relevant, and why. Second, they must explain why some uses of embryos in research fail to acknowledge what is morally relevant about the embryo, and thereby involve a violation of the moral respect that they are due. I shall show that providing such explanations may be more difficult than it first appears.


Activity of Cerebellar Nuclei Neurons Correlates with ZebrinII Identity of Their Purkinje Cell Afferents.
Purkinje cells (PCs) in the cerebellar cortex can be divided into at least two main subpopulations: one subpopulation that prominently expresses ZebrinII (Z+), and shows a relatively low simple spike firing rate, and another that hardly expresses ZebrinII (Z-) and shows higher baseline firing rates. Likewise, the complex spike responses of PCs, which are evoked by climbing fiber inputs and thus reflect the activity of the inferior olive (IO), show the same dichotomy. However, it is not known whether the target neurons of PCs in the cerebellar nuclei (CN) maintain this bimodal distribution. Electrophysiological recordings in awake adult mice show that the rate of action potential firing of CN neurons that receive input from Z+ PCs was consistently lower than that of CN neurons innervated by Z- PCs. Similar in vivo recordings in juvenile and adolescent mice indicated that the firing frequency of CN neurons correlates to the ZebrinII identity of the PC afferents in adult, but not postnatal stages. Finally, the spontaneous action potential firing pattern of adult CN neurons recorded in vitro revealed no significant differences in intrinsic pacemaking activity between ZebrinII identities. Our findings indicate that all three main components of the olivocerebellar loop, i.e., PCs, IO neurons and CN neurons, operate at a higher rate in the Z- modules.


A Genome-Wide Screen Reveals that the Vibrio cholerae Phosphoenolpyruvate Phosphotransferase System Modulates Virulence Gene Expression.
Diverse environmental stimuli and a complex network of regulatory factors are known to modulate expression of Vibrio cholerae's principal virulence factors. However, there is relatively little known about how metabolic factors impinge upon the pathogen's well-characterized cascade of transcription factors that induce expression of cholera toxin and the toxin-coregulated pilus (TCP). Here, we used a transposon insertion site (TIS) sequencing-based strategy to identify new factors required for expression of tcpA, which encodes the major subunit of TCP, the organism's chief intestinal colonization factor. Besides identifying most of the genes known to modulate tcpA expression, the screen yielded ptsI and ptsH, which encode the enzyme I (EI) and Hpr components of the V. cholerae phosphoenolpyruvate phosphotransferase system (PTS). In addition to reduced expression of TcpA, strains lacking EI, Hpr, or the associated EIIA(Glc) protein produced less cholera toxin (CT) and had a diminished capacity to colonize the infant mouse intestine. The PTS modulates virulence gene expression by regulating expression of tcpPH and aphAB, which themselves control expression of toxT, the central activator of virulence gene expression. One mechanism by which PTS promotes virulence gene expression appears to be by modulating the amounts of intracellular cyclic AMP (cAMP). Our findings reveal that the V. cholerae PTS is an additional modulator of the ToxT regulon and demonstrate the potency of loss-of-function TIS sequencing screens for defining regulatory networks.


Community nurses' perceptions of providing bereavement care.
This study explored the perceptions of bereavement support offered to clients and their carers and family by community nurses (CNs) in three community health centres located in a single area health service. In the context of an ageing population, it is pertinent to review CNs' perceptions in providing bereavement services. Early assessment and intervention is likely to prevent complicated grief occurring in the community. The bereavement support provided by CNs, considered here within a person-centred framework, enables identification of complicated grief. Semistructured interviews were held with 10 CNs and were transcribed verbatim. Transcriptions were analysed for major themes, and responses were grouped in relation to the study aims and themes emerging from the interviews. The themes discussed in this paper are as follows: the carer as a focus of palliative care; bereavement support as an outlet for carers; and the ending of the relationship between carers and CNs that is facilitated through bereavement support. The study provides evidence that supports the adoption of a model of bereavement support delivered by CNs as a means of reducing the likelihood of complicated grief occurring in the community. Further, the visits provide an important opportunity for nurses and carers to satisfactorily complete their relationship.


Suppression of superconductivity in mesoscopic superconductors.
We propose a new boundary-driven phase transition associated with vortex nucleation in mesoscopic superconductors (of size of the order of, or larger than, the penetration depth). We derive the rescaling equations and we show that boundary effects associated with vortex nucleation lower the conventional transition temperature in mesoscopic superconductors by an amount which is a function of the size of the superconductor. This result explains recent experiments in small superconductors where it was found that the transition temperature depends on the size of the system and is lower than the critical Berezinskĭ-Kosterlitz-Thouless temperature.


Effect of mixed and single crops on disease suppressiveness of soils.
ABSTRACT The effect of mixed cropping on disease suppressiveness of soils was tested for two cropping systems, Brussels sprouts-barley and triticale-white clover. Disease suppressiveness of field soils was evaluated in bioassays for the soilborne pathogens Rhizoctonia solani, Fusarium oxysporum f. sp. lini, and Gaeumannomyces graminis var. tritici. For both cropping systems, mixed cropping did not enhance disease suppressiveness of the soils. In some cases, soil cropped to barley alone was significantly more suppressive to F. oxysporum f. sp. lini than soils cropped to Brussels sprouts or the mixture of Brussels sprouts and barley. Analyses of the diversity of the indigenous bacterial and fungal microflora by denaturing gradient gel electrophoresis of amplified 16S- and 18S-rDNA fragments, respectively, revealed, in most cases, no significant differences between mixed and mono-cropped soils. In conclusion, in this study, mixed cropping of soils with Brussels sprouts and barley or with triticale and white clover did not enhance microbial diversity or disease suppressiveness of soils to three different soilborne plant pathogens.


HIV-specific cytolytic CD4 T cell responses during acute HIV infection predict disease outcome.
Early immunological events during acute HIV infection are thought to fundamentally influence long-term disease outcome. Whereas the contribution of HIV-specific CD8 T cell responses to early viral control is well established, the role of HIV-specific CD4 T cell responses in the control of viral replication after acute infection is unknown. A growing body of evidence suggests that CD4 T cells-besides their helper function-have the capacity to directly recognize and kill virally infected cells. In a longitudinal study of a cohort of individuals acutely infected with HIV, we observed that subjects able to spontaneously control HIV replication in the absence of antiretroviral therapy showed a significant expansion of HIV-specific CD4 T cell responses-but not CD8 T cell responses-compared to subjects who progressed to a high viral set point (P = 0.038). Markedly, this expansion occurred before differences in viral load or CD4 T cell count and was characterized by robust cytolytic activity and expression of a distinct profile of perforin and granzymes at the earliest time point. Kaplan-Meier analysis revealed that the emergence of granzyme A(+) HIV-specific CD4 T cell responses at baseline was highly predictive of slower disease progression and clinical outcome (average days to CD4 T cell count <350/μl was 575 versus 306, P = 0.001). These data demonstrate that HIV-specific CD4 T cell responses can be used during the earliest phase of HIV infection as an immunological predictor of subsequent viral set point and disease outcome. Moreover, these data suggest that expansion of granzyme A(+) HIV-specific cytolytic CD4 T cell responses early during acute HIV infection contributes substantially to the control of viral replication.


Maternal hormones linking maternal body mass index and dietary intake to birth weight.
Obese women often give birth to large-for-gestational age infants (typically defined as a birth weight greater than the 90th percentile), who are at risk of birth injuries and of developing metabolic syndrome later in life. The mechanisms underlying increased fetal growth remain to be established. We aimed to identify maternal hormones that can explain the link between dietary intake, body mass index (BMI), and birth weight. Pregnant women with BMIs (in kg/m(2)) ranging from 17 to 44 (n = 49) were recruited in gestational weeks 8-12. Serum hormone concentrations were measured and dietary history interviews were performed in the first and third trimesters. Multiple regression models were produced to identify hormones that correlate with birth weight and are influenced by BMI or dietary factors. We found a strong positive correlation between BMI and first- and third-trimester insulin and leptin concentrations and a negative correlation between BMI and first-trimester adiponectin and first- and third-trimester insulin-like growth factor binding protein-1 (IGFBP-1). Maternal total fat intake in the first trimester was positively correlated with maternal leptin and inversely correlated with adiponectin. In addition, third-trimester total fat intake was positively correlated with circulating resistin concentrations. First-trimester maternal serum resistin was positively correlated with birth weight, whereas third-trimester maternal IGFBP-1 was negatively correlated with birth weight. High first-trimester maternal serum resistin and low third-trimester IGFBP-1 were correlated with increased birth weight. We propose that low serum concentrations of IGFBP-1 represent a link between high BMI and increased fetal growth by increasing the bioavailability of insulin-like growth factor-I, which up-regulates placental nutrient transport.


United States State-Level Variation in the Use of Neuraxial Analgesia During Labor for Pregnant Women.
Neuraxial labor analgesia is recognized as the most effective method of providing pain relief during labor. Little is known about variation in the rates of neuraxial analgesia across US states. Identifying the presence and extent of variation may provide insights into practice variation and may indicate where access to neuraxial analgesia is inadequate. To test the hypothesis that variation exists in neuraxial labor analgesia use among US states. Retrospective, population-based, cross-sectional analysis using US birth certificate data. Participants were 2 625 950 women who underwent labor in 2015. State-specific prevalence of neuraxial analgesia per 100 women who underwent labor and variability in neuraxial analgesia use among states, assessed using multilevel multivariable regression modeling with the median odds ratio and the intraclass correlation coefficient to evaluate variation by state. In the study population of 2 625 950 women, 0.1% (n = 2010) were younger than 15 years, 7.0% (n = 183 546) were between the ages of 15 and 19 years, 23.6% (n = 620 118) were between the ages of 20 and 24 years, 29.6% (n = 777 957) were between the ages of 25 and 29 years, 26.0% (n = 683 656) were between the ages of 30 and 34 years, 11.4% (n = 298 237) were between the ages of 35 and 39 years, 2.2% (n = 57 130) were between the ages of 40 and 44 years, and 0.1% (n = 3296) were between the ages of 45 and 54 years. More than 90% were privately insured or insured with Medicaid. Neuraxial analgesia was used by 73.1% (n = 1 920 368) of women. After adjustment for antepartum, obstetric, and intrapartum factors, Maine had the lowest neuraxial analgesia prevalence (36.6%; 95% CI, 33.2%-40.1%) and Nevada the highest (80.1%; 95% CI, 78.3%-81.7%). The adjusted median odds ratio was 1.5 (95% CI, 1.4-1.6), and the intraclass correlation coefficient was 5.4% (95% CI, 4.0%-7.9%). Results of this study suggest that a small portion of the overall variation in neuraxial analgesia use is explained by US states. Unmeasured patient-level and hospital-level factors likely account for a large portion of the variation between states. Efforts should be made to understand what the main reasons are for this variation and whether the variation influences maternal or perinatal outcomes.


Synthetic lethality of cytolytic HSV-1 in cancer cells with ATRX and PML deficiency.
Cancers that utilize the alternative lengthening of telomeres (ALT) mechanism for telomere maintenance are often difficult to treat and have a poor prognosis. They are also commonly deficient for expression of ATRX protein, a repressor of ALT activity, and a component of promyelocytic leukemia nuclear bodies (PML NBs) that are required for intrinsic immunity to various viruses. Here, we asked whether ATRX deficiency creates a vulnerability in ALT cancer cells that could be exploited for therapeutic purposes. We showed in a range of cell types that a mutant herpes simplex virus type 1 (HSV-1) lacking ICP0, a protein that degrades PML NB components including ATRX, was ten- to one thousand-fold more effective in infecting ATRX-deficient cells than wild-type ATRX-expressing cells. Infection of co-cultured primary and ATRX-deficient cancer cells revealed that mutant HSV-1 selectively killed ATRX-deficient cells. Sensitivity to mutant HSV-1 infection also correlated inversely with PML protein levels, and we showed that ATRX upregulates PML expression at both the transcriptional and post-transcriptional levels. These data provide a basis for predicting, based on ATRX or PML levels, which tumors will respond to a selective oncolytic herpesvirus.


A soil-inspired dynamically responsive chemical system for microbial modulation.
Interactions between the microbiota and their colonized environments mediate critical pathways from biogeochemical cycles to homeostasis in human health. Here we report a soil-inspired chemical system that consists of nanostructured minerals, starch granules and liquid metals. Fabricated via a bottom-up synthesis, the soil-inspired chemical system can enable chemical redistribution and modulation of microbial communities. We characterize the composite, confirming its structural similarity to the soil, with three-dimensional X-ray fluorescence and ptychographic tomography and electron microscopy imaging. We also demonstrate that post-synthetic modifications formed by laser irradiation led to chemical heterogeneities from the atomic to the macroscopic level. The soil-inspired material possesses chemical, optical and mechanical responsiveness to yield write-erase functions in electrical performance. The composite can also enhance microbial culture/biofilm growth and biofuel production in vitro. Finally, we show that the soil-inspired system enriches gut bacteria diversity, rectifies tetracycline-induced gut microbiome dysbiosis and ameliorates dextran sulfate sodium-induced rodent colitis symptoms within in vivo rodent models.


Effect of a low-level laser on bone regeneration after rapid maxillary expansion.
In this study, we evaluated the effects of a low-level laser on bone regeneration in rapid maxillary expansion procedures. Twenty-seven children, aged 8 to 12 years, took part in the experiment, with a mean age of 10.2 years, divided into 2 groups: the laser group (n = 14), in which rapid maxillary expansion was performed in conjunction with laser use, and the no-laser group (n = 13), with rapid maxillary expansion only. The activation protocol of the expansion screw was 1 full turn on the first day and a half turn daily until achieving overcorrection. The laser type used was a laser diode (TWIN Laser; MMOptics, São Carlos, Brazil), according to the following protocol: 780 nm wavelength, 40 mW power, and 10 J/cm(2) density at 10 points located around the midpalatal suture. The application stages were 1 (days 1-5 of activation), 2 (at screw locking, on 3 consecutive days), 3, 4, and 5 (7, 14, and 21 days after stage 2). Occlusal radiographs of the maxilla were taken with the aid of an aluminum scale ruler as a densitometry reference at different times: T1 (initial), T2 (day of locking), T3 (3-5 days after T2), T4 (30 days after T3), and T5 (60 days after T4). The radiographs were digitized and submitted to imaging software (Image Tool; UTHSCSA, San Antonio, Tex) to measure the optic density of the previously selected areas. To perform the statistical test, analysis of covariance was used, with the time for the evaluated stage as the covariable. In all tests, a significance level of 5% (P <0.05) was adopted. From the evaluation of bone density, the results showed that the laser improved the opening of the midpalatal suture and accelerated the bone regeneration process. The low-level laser, associated with rapid maxillary expansion, provided efficient opening of the midpalatal suture and influenced the bone regeneration process of the suture, accelerating healing.


Michler's hydrol blue elucidates structural differences in prion strains.
Yeast prions provide self-templating protein-based mechanisms of inheritance whose conformational changes lead to the acquisition of diverse new phenotypes. The best studied of these is the prion domain (NM) of Sup35, which forms an amyloid that can adopt several distinct conformations (strains) that confer distinct phenotypes when introduced into cells that do not carry the prion. Classic dyes, such as thioflavin T and Congo red, exhibit large increases in fluorescence when bound to amyloids, but these dyes are not sensitive to local structural differences that distinguish amyloid strains. Here we describe the use of Michler's hydrol blue (MHB) to investigate fibrils formed by the weak and strong prion fibrils of Sup35NM and find that MHB differentiates between these two polymorphs. Quantum mechanical time-dependent density functional theory (TDDFT) calculations indicate that the fluorescence properties of amyloid-bound MHB can be correlated to the change of binding site polarity and that a tyrosine to phenylalanine substitution at a binding site could be detected. Through the use of site-specific mutants, we demonstrate that MHB is a site-specific environmentally sensitive probe that can provide structural details about amyloid fibrils and their polymorphs.


"Multi-component reactions : emerging chemistry in drug discovery" 'from xylocain to crixivan'.
With the recent emergence of combinatorial chemistry and high-speed parallel synthesis for drug discovery applications, the multi-component reaction (MCR) has seen a resurgence of interest. Easily automated one-pot reactions, such as the Ugi and Passerini reactions, are powerful tools for producing diverse arrays of compounds, often in one step and high yield. Despite this synthetic potential, the Ugi reaction is limited by producing products that are flexible and peptide-like, often being classified as 'non drug-like'. This review details developments of new, highly atom-economic MCR derived chemical methods, which enable the fast and efficient production of chemical libraries comprised of a variety of biologically relevant templates. Representative examples will also be given demonstrating the successful impact of MCR combinatorial methods at different stages of the lead discovery, lead optimization and pre-clinical process development arenas. This will include applications spanning biological tools, natural products and natural product-like diversity, traditional small molecule and 'biotech' therapeutics respectively. In particular, this review will focus on applications of isocyanide based MCR (IMCR) reactions.


Perceived dyscognition reported by patients with fibromyalgia.
Patients with fibromyalgia often report dyscognition as a symptom; however, the literature on this symptom is sparse. Our objective for this cross-sectional study was to characterize dyscognition among patients with fibromyalgia, identify comorbid symptoms associated with dyscognition, and evaluate its relation with fibromyalgia severity. Dyscognition was assessed with the Multiple Abilities Self-report Questionnaire (MASQ) for 681 patients with fibromyalgia. Other assessed comorbid symptoms were pain, fatigue, sleep problems, mood, physical and mental health, and autonomic function. Correlation and regression modeling were used to identify relations between the MASQ subscales and other fibromyalgia symptoms. Mixed analysis of variance was used to examine the profile of dyscognition in different levels of fibromyalgia. MASQ subscale scores from a previously described healthy normal control population were used for comparison. The mean (SD) age of the study patients was 55.8 (12.6) years, and most patients were female (93%) and white (91%). Perceived dyscognition was most related to depression, anxiety, and autonomic function. Across all fibromyalgia severity levels, patients had significantly higher levels of perceived dyscognition than the healthy controls. Significant differences existed for the MASQ total and most MASQ subscales among patients with mild, moderate, and severe fibromyalgia. Our study results provide further evidence that perceived dyscognition in fibromyalgia is influenced by various comorbid symptoms. In treating patients with fibromyalgia who have dyscognition, clinicians should consider the multiple types of dyscognition and the effects of other fibromyalgia symptoms.


Development of computerized adaptive testing (CAT) for the EORTC QLQ-C30 physical functioning dimension.
Computerized adaptive test (CAT) methods, based on item response theory (IRT), enable a patient-reported outcome instrument to be adapted to the individual patient while maintaining direct comparability of scores. The EORTC Quality of Life Group is developing a CAT version of the widely used EORTC QLQ-C30. We present the development and psychometric validation of the item pool for the first of the scales, physical functioning (PF). Initial developments (including literature search and patient and expert evaluations) resulted in 56 candidate items. Responses to these items were collected from 1,176 patients with cancer from Denmark, France, Germany, Italy, Taiwan, and the United Kingdom. The items were evaluated with regard to psychometric properties. Evaluations showed that 31 of the items could be included in a unidimensional IRT model with acceptable fit and good content coverage, although the pool may lack items at the upper extreme (good PF). There were several findings of significant differential item functioning (DIF). However, the DIF findings appeared to have little impact on the PF estimation. We have established an item pool for CAT measurement of PF and believe that this CAT instrument will clearly improve the EORTC measurement of PF.


Tbx20 dose-dependently regulates transcription factor networks required for mouse heart and motoneuron development.
To elucidate the function of the T-box transcription factor Tbx20 in mammalian development, we generated a graded loss-of-function series by transgenic RNA interference in entirely embryonic stem cell-derived mouse embryos. Complete Tbx20 knockdown resulted in defects in heart formation, including hypoplasia of the outflow tract and right ventricle, which derive from the anterior heart field (AHF), and decreased expression of Nkx2-5 and Mef2c, transcription factors required for AHF formation. A mild knockdown led to persistent truncus arteriosus (unseptated outflow tract) and hypoplastic right ventricle, entities similar to human congenital heart defects, and demonstrated a critical requirement for Tbx20 in valve formation. Finally, an intermediate knockdown revealed a role for Tbx20 in motoneuron development, specifically in the regulation of the transcription factors Isl2 and Hb9, which are important for terminal differentiation of motoneurons. Tbx20 could activate promoters/enhancers of several genes in cultured cells, including the Mef2c AHF enhancer and the Nkx2-5 cardiac enhancer. The Mef2c AHF enhancer relies on Isl1- and Gata-binding sites. We identified a similar Isl1 binding site in the Nkx2-5 AHF enhancer, which in transgenic mouse embryos was essential for activity in a large part of the heart, including the outflow tract. Tbx20 synergized with Isl1 and Gata4 to activate both the Mef2c and Nkx2-5 enhancers, thus providing a unifying mechanism for gene activation by Tbx20 in the AHF. We conclude that Tbx20 is positioned at a critical node in transcription factor networks required for heart and motoneuron development where it dose-dependently regulates gene expression.


Effects of LSD on Ca++ currents in central 5-HT-containing neurons: 5-HT1A receptors may play a role in hallucinogenesis.
Drugs that influence the activity of central serotonergic neurons by activating a 5-hydroxytryptamine subtype of receptor (5-HT1A) alter mood and perception. Previously, we demonstrated with whole-cell recordings from acutely isolated 5-HT-containing dorsal raphe (DR) neurons from the adult rat that 5-HT inhibited Ca++ current and activated K+ current in DR neurons. We now show that D-lysergic acid diethylamide (LSD) mimics the actions of 5-HT; it dramatically suppresses Ca++ current in a dose-dependent manner and activates an inwardly rectifying K+ conductance. Spiperone (0.2 microM), a 5-HT1A/5-HT2 antagonist, blocks the effect of both LSD and 5-HT. The nonhallucinogenic structural analog 2-bromo-LSD (2-Bol) at 10 microM has no effect on either Ca++ or K+ current by itself, but it competitively antagonizes both effects of LSD. Inhibition of 5-HT release resulting from 5-HT1A receptor activation may play an integral role in the hallucinogenic actions of LSD by reducing competition between 5-HT and LSD for the postsynaptic 5-HT receptors.


Comparative gene expression between two yeast species.
Comparative genomics brings insight into sequence evolution, but even more may be learned by coupling sequence analyses with experimental tests of gene function and regulation. However, the reliability of such comparisons is often limited by biased sampling of expression conditions and incomplete knowledge of gene functions across species. To address these challenges, we previously systematically generated expression profiles in Saccharomyces bayanus to maximize functional coverage as compared to an existing Saccharomyces cerevisiae data repository. In this paper, we take advantage of these two data repositories to compare patterns of ortholog expression in a wide variety of conditions. First, we developed a scalable metric for expression divergence that enabled us to detect a significant correlation between sequence and expression conservation on the global level, which previous smaller-scale expression studies failed to detect. Despite this global conservation trend, between-species gene expression neighborhoods were less well-conserved than within-species comparisons across different environmental perturbations, and approximately 4% of orthologs exhibited a significant change in co-expression partners. Furthermore, our analysis of matched perturbations collected in both species (such as diauxic shift and cell cycle synchrony) demonstrated that approximately a quarter of orthologs exhibit condition-specific expression pattern differences. Taken together, these analyses provide a global view of gene expression patterns between two species, both in terms of the conditions and timing of a gene's expression as well as co-expression partners. Our results provide testable hypotheses that will direct future experiments to determine how these changes may be specified in the genome.


Did the English strategy reduce inequalities in health? A difference-in-difference analysis comparing England with three other European countries.
Between 1997 and 2010, the English government pursued an ambitious programme to reduce health inequalities, the explicit and sustained commitment of which was historically and internationally unique. Previous evaluations have produced mixed results. None of these evaluations have, however, compared the trends in health inequalities within England with those in other European countries. We carried out an innovative analysis to assess whether changes in trends in health inequalities observed in England after the implementation of its programme, have been more favourable than those in other countries without such a programme. Data were obtained from nationally representative surveys carried out in England, Finland, the Netherlands and Italy for years around 1990, 2000 and 2010. A modified difference-in-difference approach was used to assess whether trends in health inequalities in 2000-2010 were more favourable as compared to the period 1990-2000 in England, and the changes in trends in inequalities after 2000 in England were then compared to those in the three comparison countries. Health outcomes were self-assessed health, long-standing health problems, smoking status and obesity. Education was used as indicator of socioeconomic position. After the implementation of the English strategy, more favourable trends in some health indicators were observed among low-educated people, but trends in health inequalities in 2000-2010 in England were not more favourable than those observed in the period 1990-2000. For most health indicators, changes in trends of health inequalities after 2000 in England were also not significantly different from those seen in the other countries. In this rigorous analysis comparing trends in health inequalities in England both over time and between countries, we could not detect a favourable effect of the English strategy. Our analysis illustrates the usefulness of a modified difference-in-difference approach for assessing the impact of policies on population-level health inequalities.


Inferring connectivity of genetic regulatory networks using information-theoretic criteria.
Recently, the concept of mutual information has been proposed for inferring the structure of genetic regulatory networks from gene expression profiling. After analyzing the limitations of mutual information in inferring the gene-to-gene interactions, this paper introduces the concept of conditional mutual information and based on it proposes two novel algorithms to infer the connectivity structure of genetic regulatory networks. One of the proposed algorithms exhibits a better accuracy while the other algorithm excels in simplicity and flexibility. By exploiting the mutual information and conditional mutual information, a practical metric is also proposed to assess the likeliness of direct connectivity between genes. This novel metric resolves a common limitation associated with the current inference algorithms, namely the situations where the gene connectivity is established in terms of the dichotomy of being either connected or disconnected. Based on the data sets generated by synthetic networks, the performance of the proposed algorithms is compared favorably relative to existing state-of-the-art schemes. The proposed algorithms are also applied on realistic biological measurements, such as the cutaneous melanoma data set, and biological meaningful results are inferred.


Enzymatic synthesis of β-xylosyl-oligosaccharides by transxylosylation using two β-xylosidases of glycoside hydrolase family 3 from Aspergillus nidulans FGSC A4.
Two β-xylosidases of glycoside hydrolase family 3 (GH 3) from Aspergillus nidulans FGSC A4, BxlA and BxlB were produced recombinantly in Pichia pastoris and secreted to the culture supernatants in yields of 16 and 118 mg/L, respectively. BxlA showed about sixfold higher catalytic efficiency (k(cat)/K(m)) than BxlB towards para-nitrophenyl β-D-xylopyranoside (pNPX) and β-1,4-xylo-oligosaccharides (degree of polymerisation 2-6). For both enzymes k(cat)/K(m) decreased with increasing β-1,4-xylo-oligosaccharide chain length. Using pNPX as donor with 9 monosaccharides, 7 disaccharides and two sugar alcohols as acceptors 18 different β-xylosyl-oligosaccharides were synthesised in 2-36% (BxlA) and 6-66% (BxlB) yields by transxylosylation. BxlA utilised the monosaccharides D-mannose, D-lyxose, D-talose, D-xylose, D-arabinose, L-fucose, D-glucose, D-galactose and D-fructose as acceptors, whereas BxlB used the same except for D-lyxose, D-arabinose and L-fucose. BxlB transxylosylated the disaccharides xylobiose, lactulose, sucrose, lactose and turanose in upto 35% yield, while BxlA gave inferior yields on these acceptors. The regioselectivity was acceptor dependent and primarily involved β-1,4 or 1,6 product linkage formation although minor products with different linkages were also obtained. Five of the 18 transxylosylation products obtained from D-lyxose, D-galactose, turanose and sucrose (two products) as acceptors were novel xylosyl-oligosaccharides, β-D-Xylp-(1→4)-D-Lyxp, β-D-Xylp-(1→6)-D-Galp, β-D-Xylp-(1→4)-α-D-Glcp-(1→3)-β-D-Fruf, β-D-Xylp-(1→4)-α-D-Glcp-(1→2)-β-D-Fruf, and β-D-Xylp-(1→6)-β-D-Fruf-(2→1)-α-D-Glcp, as structure-determined by 2D NMR, indicating that GH3 β-xylosidases are able to transxylosylate a larger variety of carbohydrate acceptors than earlier reported. Furthermore, transxylosylation of certain acceptors resulted in mixtures. Some of these products are also novel, but the structures of the individual products could not be determined.


Mutation analysis of the coding sequence of the MECP2 gene in infantile autism.
Mutations in the coding region of the methyl-CpG-binding protein 2 ( MECP2) gene cause Rett syndrome and have also been reported in a number of X-linked mental retardation syndromes. Furthermore, such mutations have recently been described in a few autistic patients. In this study, a large sample of individuals with autism was screened in order to elucidate systematically whether specific mutations in MECP2 play a role in autism. The mutation analysis of the coding sequence of the gene was performed by denaturing high-pressure liquid chromatography and direct sequencing. Taken together, 14 sequence variants were identified in 152 autistic patients from 134 German families and 50 unrelated patients from the International Molecular Genetic Study of Autism Consortium affected relative-pair sample. Eleven of these variants were excluded for having an aetiological role as they were either silent mutations, did not cosegregate with autism in the pedigrees of the patients or represented known polymorphisms. The relevance of the three remaining mutations towards the aetiology of autism could not be ruled out, although they were not localised within functional domains of MeCP2 and may be rare polymorphisms. Taking into account the large size of our sample, we conclude that mutations in the coding region of MECP2 do not play a major role in autism susceptibility. Therefore, infantile autism and Rett syndrome probably represent two distinct entities at the molecular genetic level.


Harnessing the cross-talk between tumor cells and tumor-associated macrophages with a nano-drug for modulation of glioblastoma immune microenvironment.
Glioblastoma (GBM) is the most frequent and malignant brain tumor with a high mortality rate. The presence of a large population of macrophages (Mφ) in the tumor microenvironment is a prominent feature of GBM and these so-called tumor-associated Mφ (TAM) closely interact with the GBM cells to promote the survival, progression and therapy resistance of the GBM. Various therapeutic strategies have been devised either targeting the GBM cells or the TAM but few have addressed the cross-talks between the two cell populations. The present study was carried out to explore the possibility of exploiting the cross-talks between the GBM cells (GC) and TAM for modulation of the GBM microenvironment through using Nano-DOX, a drug composite based on nanodiamonds bearing doxorubicin. In the in vitro work on human cell models, Nano-DOX-loaded TAM were first shown to be viable and able to infiltrate three-dimensional GC spheroids and release cargo drug therein. GC were then demonstrated to encourage Nano-DOX-loaded TAM to unload Nano-DOX back into GC which consequently emitted damage-associated molecular patterns (DAMPs) that are powerful immunostimulatory agents as well as indicators of cell damage. Nano-DOX was next proven to be a more potent inducer of GC DAMPs emission than doxorubicin. As a result, Nano-DOX-damaged GC exhibited an enhanced ability to attract both TAM and Nano-DOX-loaded TAM. Most remarkably, Nano-DOX-damaged GC reprogrammed the TAM from a pro-GBM phenotype to an anti-GBM phenotype that suppressed GC growth. Finally, the in vivo relevance of the in vitro findings was tested in animal study. Mice bearing orthotopic human GBM xenografts were intravenously injected with Nano-DOX-loaded mouse TAM which were found releasing drug in the GBM xenografts 24h after injection. GC damage was evidenced by the induction of DAMPs emission within the xenografts and a shift of TAM phenotype was detected as well. Taken together, our results demonstrate a novel way with therapeutic potential to harness the cross-talk between GBM cells and TAM for modulation of the tumor immune microenvironment.


CD25 Blockade Delays Regulatory T Cell Reconstitution and Does Not Prevent Graft-versus-Host Disease After Allogeneic Hematopoietic Cell Transplantation.
Daclizumab, a humanized monoclonal antibody, binds CD25 and blocks formation of the IL-2 receptor on T cells. A study of daclizumab as acute graft-versus-host disease (GVHD) prophylaxis after unrelated bone marrow transplantation was conducted before the importance of CD25+FOXP3+ regulatory T cells (Tregs) was recognized. Tregs can abrogate the onset of GVHD. The relation between Tregs and a graft-versus-malignancy effect is not fully understood. An international, multicenter, double-blind clinical trial randomized 210 adult or pediatric patients to receive 5 weekly doses of daclizumab at 0.3 mg/kg (n = 69) or 1.2 mg/kg (n = 76) or placebo (n = 65) after unrelated marrow transplantation for treatment of hematologic malignancies or severe aplastic anemia. The risk of acute GVHD did not differ among the groups (P = .68). Long-term follow-up of clinical outcomes and correlative analysis of peripheral blood T cell phenotype suggested that the patients treated with daclizumab had an increased risk of chronic GVHD (hazard ratio [HR], 1.49; 95% confidence interval [CI], 1.0 to 2.3; P = .08) and a decreased risk of relapse (HR, 0.57; 95% CI, 0.3 to 1.0; P = .05), but similar survival (HR, 0.89; 95% CI, 0.6 to 1.3; P = .53). T cells from a subset of patients (n = 107) were analyzed by flow cytometry. Compared with placebo, treatment with daclizumab decreased the proportion of Tregs among CD4 T cells at days 11-35 and increased the proportion of central memory cells among CD4 T cells at 1 year. Prophylactic administration of daclizumab does not prevent acute GVHD, but may increase the risk of chronic GVHD and decrease the risk of relapse. By delaying Treg reconstitution and promoting immunologic memory, anti-CD25 therapy may augment alloreactivity and antitumor immunity.


Association of Catastrophic Neonatal Outcomes With Increased Rate of Subsequent Cesarean Deliveries.
To evaluate whether full-term deliveries resulting in neonates diagnosed with hypoxic-ischemic encephalopathy are associated with a significant increase in the rate of subsequent unscheduled cesarean deliveries. We conducted a retrospective chart review study and examined all deliveries in the Department of Obstetrics and Gynecology at Hadassah University Hospital, Mt. Scopus campus, Jerusalem, Israel, during 2009-2014. We reviewed all cases of hypoxic-ischemic encephalopathy in singleton, term, liveborn neonates and identified seven such cases, three of which were attributed to obstetric mismanagement and four that were not. We measured the rate of unscheduled cesarean deliveries before and after the events and their respective hazard ratio. Before a mismanaged delivery resulting in hypoxic-ischemic encephalopathy, the baseline rate of unscheduled cesarean deliveries was approximately 80 unscheduled cesarean deliveries for every 1,000 deliveries. In the first 4 weeks immediately after each of the three identified cases, there was a significant increase in the rate of unscheduled cesarean deliveries by an additional 48 unscheduled cesarean deliveries per 1,000 deliveries (95% confidence interval [CI] 27-70/1,000). This increase was transient and lasted approximately 4 weeks. We estimated that each case was associated with approximately 17 additional unscheduled cesarean deliveries (95% CI 8-27). There was no increase in the rate of unscheduled cesarean deliveries in cases of hypoxic-ischemic encephalopathy that were not associated with mismanagement. The increase in the rate of unscheduled cesarean deliveries after a catastrophic neonatal outcome may result in short-term changes in obstetricians' risk evaluation.


Characterization of the early endosome and putative endocytic carrier vesicles in vivo and with an assay of vesicle fusion in vitro.
We have investigated two aspects of membrane traffic at early stages of endocytosis: membrane fusion and microtubule-dependent transport. As a marker, we have used the trans-membrane glycoprotein G of vesicular stomatitis virus implanted into the plasma membrane and then internalized for different times at 37 degrees C. The corresponding endosomal fractions were immunoisolated using the cytoplasmic domain of the G protein as antigen. These fractions were then used in an in vitro assay to quantify the efficiency of fusion between endosomal vesicles. To identify the vesicular partners of the fusion, these in vitro studies were combined with in vivo biochemical and morphological experiments. Internalized molecules were delivered to early endosomal elements, which corresponded to a network of tubular and tubulovesicular structures. Rapid recycling back to the plasma membrane and routing to late stages of the pathway occurred from these early endosomal elements. These elements exhibited a high and specific fusion activity with each other in vitro, suggesting that individual elements of the early endosomal compartment interact with each other in vivo. After their appearance in the early endosome, the molecules destined to be degraded were observed at the next stage of the pathway in distinct spherical vesicles (0.5 micron diam) and then in late endosomes and lysosomes. When the microtubules were depolymerized with nocodazole, endocytosis proceeded as in control cells. However, internalized molecules remained in the spherical vesicles and did not appear in late endosomes or lysosomes. These spherical vesicles had relatively little fusion activity with each other or with early endosomal elements in vitro. Our observations suggest that the spherical vesicles mediate transport between the early endosome and late endosomes and that this process requires intact microtubules.


The effect of mevalonic acid deprivation on enzymes of DNA replication in cells emerging from quiescence.
We have investigated the biochemical basis of the mevalonate dependence of DNA replication. Stimulating quiescent rat hepatoma cells to proliferate in the presence of compactin, an inhibitor of mevalonate synthesis, prevented DNA replication in as many as 80% of these cells. The percentage of cells that failed to replicate DNA increased with the increased duration of quiescence. Aphidicolin-sensitive DNA polymerase and ornithine decarboxylase activities were selectively decreased in compactin-treated cells, whereas RNA and protein synthesis, the level of dihydrofolate reductase and aphidicolin-resistant DNA polymerase activity were unaffected. Adding putrescine, the product of ornithine decarboxylase and the precursor of other polyamines, did not restore DNA replication. Our results demonstrate that the decreased activities of at least two DNA-replication enzymes are among the proximal causes of the failure of mevalonate-deprived cells to synthesize DNA. More importantly, our data indicate that a mevalonate-dependent factor(s) is progressively depleted during quiescence, and that inability to resynthesize this factor(s) may be the ultimate cause of the failure of resting cells to replicate DNA when stimulated to proliferate in the absence of mevalonate.


Effect of Jianpiyiqi granule on ocular myasthenia gravis in children prepared with a formula from Traditional Chinese Medicine.
To investigate the effect of Jianpiyiqi granule, prepared according to a Traditional Chinese Medicine (TCM) formula, on the ocular type of myasthenia gravis (MG) caused by a defect in synaptic transmission at the neuromuscular junction. A total of 155 children with ocular MG were recruited from January 2008 to January 2015. All individuals received ineffective glucocorticoid treatment prior to admission, and were given Jianpiyiqi granules (two doses per day) for 12 months. Plasma levels of acetylcholine receptor autoantibodies (AChR-Ab), cytokines, immune parameters and clinical score were analyzed. After intervention with Jianpiyiqi granule for 12 months, the levels of cytokines [including interleukin (IL)-6, IL-12, IL-17], immune parameters [including immunoglobulin G (IgG), IgM, free triiodothyronine, free thyroxine] and AChR-Ab were significantly decreased (P = 0.042, P = 0.049, P = 0.011, P = 0.02, P = 0.04, P = 0.03, P = 0.01; F = 21.60, P = 0.000). In contrast, IL-4 levels were significantly increased (P = 0.049). The absolute clinical score after treatment declined significantly compared with before treatment (F = 33.24, P = 0.000). The effective cure rate and the total effective rate gradually increased to a maximum of 149 (96.2%) and 151 (97.4%) in MG patients after 12 months of treatment, respectively. The Jianpiyiqi granule treatment lowered AChR-Ab levels and improved cytokine and other immune parameter levels, which suggests that the granule could be an ancillary treatment for ocular MG in children caused by a defect in synaptic transmission at the neuromuscular junction.


Genetic influences on political ideologies: twin analyses of 19 measures of political ideologies from five democracies and genome-wide findings from three populations.
Almost 40 years ago, evidence from large studies of adult twins and their relatives suggested that between 30 and 60% of the variance in social and political attitudes could be explained by genetic influences. However, these findings have not been widely accepted or incorporated into the dominant paradigms that explain the etiology of political ideology. This has been attributed in part to measurement and sample limitations, as well the relative absence of molecular genetic studies. Here we present results from original analyses of a combined sample of over 12,000 twins pairs, ascertained from nine different studies conducted in five democracies, sampled over the course of four decades. We provide evidence that genetic factors play a role in the formation of political ideology, regardless of how ideology is measured, the era, or the population sampled. The only exception is a question that explicitly uses the phrase "Left-Right". We then present results from one of the first genome-wide association studies on political ideology using data from three samples: a 1990 Australian sample involving 6,894 individuals from 3,516 families; a 2008 Australian sample of 1,160 related individuals from 635 families and a 2010 Swedish sample involving 3,334 individuals from 2,607 families. No polymorphisms reached genome-wide significance in the meta-analysis. The combined evidence suggests that political ideology constitutes a fundamental aspect of one's genetically informed psychological disposition, but as Fisher proposed long ago, genetic influences on complex traits will be composed of thousands of markers of very small effects and it will require extremely large samples to have enough power in order to identify specific polymorphisms related to complex social traits.


Criminalising consensual sexual behaviour in the context of HIV: consequences, evidence, and leadership.
This paper provides an overview of the use of the criminal law to regulate sexual behaviour in three areas of critical importance: (1) HIV exposure in otherwise consensual sex, (2) sex work and (3) sexual activity largely affecting sexual minorities. It analyses criminal law pertaining to these three distinct areas together, allowing for a more comprehensive and cohesive understanding of criminalisation and its effects. The paper highlights current evidence of how criminalisation undermines HIV prevention and treatment. It focuses on three specific negative effects of criminalisation: (1) enhancing stigma and discrimination, (2) undermining public health intervention through legal marginalisation and (3) placing people in state custody. The paper also highlights gaps in evidence and the need for strong institutional leadership from UN agencies in ending the criminalisation of consensual sexual activity. This paper serves two goals: (1) highlighting the current state of research and emphasising where key institutions have or have not provided appropriate leadership on these issues and (2) establishing a forward-looking agenda that includes a concerted response to the inappropriate use of the criminal law with respect to sexuality as part of the global response to HIV.


Cytosolic expression of SecA2 is a prerequisite for long-term protective immunity.
Induction of efficient adaptive T cell-mediated immunity against the intracellular bacterium Listeria monocytogenes requires its successful invasion of host cell cytosol. However, it is not clear whether its cytosolic escape and growth are sufficient to induce T cell-mediated clearance and protection upon secondary infection. To investigate this issue, we have searched for mutants that do not induce long-term protective immunity yet invade the cytosol of infected cells. We found that mice immunized with L. monocytogenes lacking the SecA2 ATPase, an auxiliary protein secretion system present in several Gram-positive pathogenic bacteria, mounted a robust cytolytic IFN-gamma-secreting CD8+ T cell response but were not protected against a secondary challenge with wild-type (wt) bacteria. Furthermore, CD8+ T cells from mice immunized with secA2- bacteria failed to transfer protection when injected into recipient mice demonstrating that they were unable to confer protection. Also, secA2- and wt L. monocytogenes spread to the same myeloid-derived cell types in vivo and SecA2 deficiency does not interfere with intracytosolic bacteria multiplication. Therefore, cytosol invasion is not sufficient for inducing secondary protective responses and induction of memory CD8+ T cells mediating long-term antibacterial protective immunity is dependent upon SecA2 expression inside the cytosol of host cells in vivo.


Localization of atherosclerotic lesions in three cerebral arterial bifurcations.
The localization of macroscopic atherosclerotic lesions have been studied in the basilar bifurcation and the left and right internal carotid bifurcations in humans. The distribution of lesions was separately assessed by a computer technique and results were displayed as contour lines. Lesions are most frequent on the outer walls of the bifurcations and on the inner curvature of the arterial bends. The material was processed for histology, and computerised morphometry was used to study the interrelationship of microscopic atheromatous lesions and intimal thickening. The percentage of the specimens with lesions around the circumference of the arteries was calculated. The peaks and troughs The peaks and troughs of histological lesions correspond exactly to the maximum and minimum contours of the macroscopic studies. Results from the microscopic examination indicate that intimal thickening and atherosclerosis are closely related processes. However, they cannot support the view that focal pad-like areas of intimal thickening are particular precursors of atheroma.


Network assessment of demethylation treatment in melanoma: Differential transcriptome-methylome and antigen profile signatures.
In melanoma, like in other cancers, both genetic alterations and epigenetic underlie the metastatic process. These effects are usually measured by changes in both methylome and transcriptome profiles, whose cross-correlation remains uncertain. We aimed to assess at systems scale the significance of epigenetic treatment in melanoma cells with different metastatic potential. Treatment by DAC demethylation with 5-Aza-2'-deoxycytidine of two melanoma cell lines endowed with different metastatic potential, SKMEL-2 and HS294T, was performed and high-throughput coupled RNA-Seq and RRBS-Seq experiments delivered differential profiles (DiP) of both transcriptomes and methylomes. Methylation levels measured at both TSS and gene body were studied to inspect correlated patterns with wide-spectrum transcript abundance levels quantified in both protein coding and non-coding RNA (ncRNA) regions. The DiP were then mapped onto standard bio-annotation sources (pathways, biological processes) and network configurations were obtained. The prioritized associations for target identification purposes were expected to elucidate the reprogramming dynamics induced by the epigenetic therapy. The interactomic connectivity maps of each cell line were formed to support the analysis of epigenetically re-activated genes. i.e. those supposedly silenced by melanoma. In particular, modular protein interaction networks (PIN) were used, evidencing a limited number of shared annotations, with an example being MAPK13 (cascade of cellular responses evoked by extracellular stimuli). This gene is also a target associated to the PANDAR ncRNA, therapeutically relevant because of its aberrant expression observed in various cancers. Overall, the non-metastatic SKMEL-2 map reveals post-treatment re-activation of a richer pathway landscape, involving cadherins and integrins as signatures of cell adhesion and proliferation. Relatively more lncRNAs were also annotated, indicating more complex regulation patterns in view of target identification. Finally, the antigen maps matched to DiP display other differential signatures with respect to the metastatic potential of the cell lines. In particular, as demethylated melanomas show connected targets that grow with the increased metastatic potential, also the potential target actionability seems to depend to some degree on the metastatic state. However, caution is required when assessing the direct influence of re-activated genes over the identified targets. In light of the stronger treatment effects observed in non-metastatic conditions, some limitations likely refer to in silico data integration tools and resources available for the analysis of tumor antigens. Demethylation treatment strongly affects early melanoma progression by re-activating many genes. This evidence suggests that the efficacy of this type of therapeutic intervention is potentially high at the pre-metastatic stages. The biomarkers that can be assessed through antigens seem informative depending on the metastatic conditions, and networks help to elucidate the assessment of possible targets actionability.