[Lambert-Eaton myasthenic syndrome associated with pulmonary squamous cell carcinoma and circulating anti-P/Q-type voltage-gated calcium channel antibody].
We report a 64-year-old man diagnosed with Lambert-Eaton myasthenic syndrome (LEMS) associated with pulmonary squamous cell carcinoma. Circulating anti-P/Q-type voltage-gated calcium channel (VGCC) antibody was detected, and the patient was treated with 3,4-diaminopyridine. At age 61, chest radiograph revealed a tumor shadow in the right upper lung field. This was surgically removed, and a histological diagnosis of moderately differentiated pulmonary squamous cell carcinoma was obtained. After about 1 year, mediastinal metastasis was detected and 5-FU was administered. Eight months later, metastasis was noted in the left frontal hemisphere, and radiosurgical therapy was performed. The brain tumor gradually shrank but generalized fatigue, thirst, and gait disturbance developed after 4 months. A diagnosis of LEMS was made on the basis of neurological findings including proximal muscle weakness and absent tendon reflexes; autonomic symptoms (thirst, constipation, and impotence); characteristic electromyographic findings; and circulating anti-P/Q-type VGCC antibody. He has been treated with 3,4-diaminopyridine at a dose of 30 mg/day, resulting in marked improvement in symptoms but little change in electromyographic findings. The present case is very rare and suggests that anti-P/Q-type VGCC antibody may be involved in the mechanism of LEMS associated with pulmonary squamous cell carcinoma.


A Nationwide Cross-Sectional Survey of UK Breast Surgeons' Views on the Management of Ductal Carcinoma In Situ.
Background. There is wide variation in the management of Ductal Carcinoma In Situ (DCIS) nationwide. We aimed to investigate whether the attitudes of surgeons towards different aspects of DCIS treatment varied by seniority of surgeon or by geographical region within the UK. Materials and Methods. A nationwide online survey targeted at UK breast surgeons was undertaken. The anonymous survey contained questions regarding demographics of respondents and specific questions regarding DCIS management that were identified as areas of uncertainty during a systematic search of the literature. Results. Responses from 80 surgeons were obtained. Approximately 57% were male and the majority were consultant or specialist registrar. Approximately 63% of participants were based in district general hospitals with all training deaneries represented. Surgeons' views on the prognosis and management of DCIS varied geographically across the UK and terminology for DCIS varied with surgeon seniority. Surgeons' views particularly differed from national guidance on indications for SLNB, tamoxifen, and follow-up practice. Conclusion. Our survey reaffirms that, irrespective of national guidelines and attempts at uniformity, there continues to be a wide variety of views amongst breast surgeons regarding the ideal management of DCIS. However, by quantifying this variation, it may be possible to take it into account when examining long-term trends in nationwide treatment data.


A statistical model to identify differentially expressed proteins in 2D PAGE gels.
Two dimensional polyacrylamide gel electrophoresis (2D PAGE) is used to identify differentially expressed proteins and may be applied to biomarker discovery. A limitation of this approach is the inability to detect a protein when its concentration falls below the limit of detection. Consequently, differential expression of proteins may be missed when the level of a protein in the cases or controls is below the limit of detection for 2D PAGE. Standard statistical techniques have difficulty dealing with undetected proteins. To address this issue, we propose a mixture model that takes into account both detected and non-detected proteins. Non-detected proteins are classified either as (a) proteins that are not expressed in at least one replicate, or (b) proteins that are expressed but are below the limit of detection. We obtain maximum likelihood estimates of the parameters of the mixture model, including the group-specific probability of expression and mean expression intensities. Differentially expressed proteins can be detected by using a Likelihood Ratio Test (LRT). Our simulation results, using data generated from biological experiments, show that the likelihood model has higher statistical power than standard statistical approaches to detect differentially expressed proteins. An R package, Slider (Statistical Likelihood model for Identifying Differential Expression in R), is freely available at http://www.cebl.auckland.ac.nz/slider.php.


How ions in solution can change the sign of the critical Casimir potential.
We show that hydrophilic ions present in a confined, near-critical aqueous mixture can lead to an attraction between like charge surfaces with opposing preferential adsorption of the two species of the mixture, even though the corresponding Casimir potential in uncharged systems is repulsive. This prediction agrees with a recent experiment [Nellen et al., Soft Matter, 2011, 7, 5360]. We also show that oppositely charged hydrophobic surfaces can repel each other, although the Casimir potential between uncharged surfaces with like preferential adsorption (selectivity) is attractive. This behavior is expected when the electrostatic screening length is larger than the correlation length, and one of the confining surfaces is strongly selective and weakly charged, whereas the other confining surface is weakly selective and strongly charged. The Casimir potential can change sign because the hydrophilic ions near the weakly hydrophobic surface can overcompensate the effect of hydrophobicity, and this surface can act as a hydrophilic one. We also predict a more attractive interaction between charged, hydrophilic surfaces and a more repulsive interaction between charged, hydrophobic surfaces than given by the sum of the Casimir and Debye-Hückel potentials. Our theory is derived systematically from a microscopic approach, and combines the Landau-type and Debye-Hückel theories with an additional contribution of an entropic origin.


Control of Oocyte Development in Cockroaches.
Development of ovarian eggs and secretory activity of colleterial glands are inhibited by the oötheca in the oöthecal chamber of Blattella germanica (L.) and by the oötheca in the uterus of Pycnoscelus surinamensis (L.). The inhibition is due to nervous stimuli from pressure of the oötheca. Removing the oötheca or severing the ventral nerve cord eliminates inhibition of the corpora allata and results in premature development of the oöcytes and resumption of activity of the colleterial glands.


Phyllodes tumor of the breast: an update.
Phyllodes tumor is an uncommon biphasic breast tumor, with the ability to recur and metastasize, and it behaves biologically like a stromal neoplasm. Traditionally, phyllodes tumors are graded by the use of a set of histologic data into benign, borderline, and malignant. In most series, all phyllodes tumors may recur, but only the borderline and malignant phyllodes tumors metastasize. On the basis of histologic features, prediction of behavior is difficult. The expression of many biological markers, including p53, hormone receptors, proliferation markers, angiogenesis group of markers, c-kit, CD10 and epidermal growth factor receptor have been explored, and many have been shown to be variably expressed, depending on the grade of the tumor. These markers are, however, of limited value in predicting the behavior of the tumor. Recently investigators have reported a plethora of genetic changes in phyllodes tumors, the most consistent of which seems to be 1q gain by comparative genomic hybridization. Some candidate genes have been mapped to various sites, and preliminary data suggest that some of these changes may be related to recurrence. It is foreseeable that more exciting data will be generated to help us to understand the etiology and pathogenesis of phyllodes tumor.


Genetic heterogeneity of HIV type 1 subtypes in Kimpese, rural Democratic Republic of Congo.
A relatively low and stable seroprevalence of HIV-1 was previously reported among pregnant women attending for antenatal care between 1988 and 1993 in Kimpese, a rural town in the Democratic Republic of Congo (DRC, formerly Zaire). To characterize the HIV-1 subtypes circulating in this area, we have examined a 330-bp fragment of the p17 region of the gag gene of HIV-1 strains obtained from 70 patients (55 mothers, 15 children), of whom 61 were epidemiologically unlinked. Phylogenetic analyses revealed the existence of at least seven HIV-1 subtypes within the Kimpese region. Among the 61 epidemiologically unlinked patients, subtype A was predominant and found in 29 (47.5%) individuals. Other subtypes cocirculating in this rural part of DRC include subtypes C (1.6%), D (9.8%), F (3.2%), G (6.5%), H (21.3%), and J (4.9%). Sequences from four patients did not cluster with any of the currently documented HIV-1 subtypes, in analyses of fragments of both the gag (247 to 330 bp, 197 bp, and 310 bp) and env (340 bp) genes. Overall, comparisons of the gag(p17) gene regions revealed high pairwise divergences (mean, 19.9%; range, 1 to 46%). This level of gag(p17) gene variation in the DRC is considerably greater than previously appreciated. These results are relevant for the molecular epidemiology of HIV-1 in Africa and for the design of a future vaccine against HIV-1 in this region.


Association of High-Mobility Group Box-1 With Th Cell-Related Cytokines in the Vitreous of Ocular Sarcoidosis Patients.
High-mobility group box-1 (HMGB1) is a nonhistone DNA-binding nuclear protein released from necrotic cells, which is also secreted by activated leukocytes and acts as a primary proinflammatory cytokine. In this study, we compared vitreous HMGB1 levels in ocular sarcoidosis with those in noninflammatory vitreoretinal diseases and evaluated its association with Th cell-related and proinflammatory cytokines. The study group consisted of 24 patients with ocular sarcoidosis. The control group consisted of 27 patients with proliferative diabetic retinopathy (PDR) and 24 with idiopathic epiretinal membrane (ERM). Vitreous fluid samples were obtained at the beginning of vitrectomy. Vitreous levels of HMGB1 and IL-1β, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IFN-γ, soluble CD40 ligand (sCD40L), and TNFα were measured. High-mobility group box-1 was detected in the vitreous of 23 of 24 patients (95.8%) with ocular sarcoidosis. Mean vitreous level of HMGB1 was the highest in the sarcoidosis group, followed by the PDR and ERM groups, with significant differences between the three groups. In the sarcoidosis group, vitreous levels of IL-6, IL-10, IL-31, IFN-γ, sCD40L, and TNFα were significantly higher than those in the idiopathic ERM group, and IFN-γ and sCD40L were significantly higher than those in the PDR group. Vitreous HMGB-1 level correlated significantly with IL-10, IFN-γ, and sCD40L levels but not with IL-6, IL-17, IL-31, or TNFα levels. The vitreous level of HMGB1 is elevated in ocular sarcoidosis and is associated with vitreous levels of Th1- and regulatory T-related cytokines, but not with proinflammatory or Th17-related cytokines.


Health effects of long-term exposure to insecticide-treated mosquito nets in the control of malaria in endemic regions, revised.
The endemicity of malaria in tropical areas of the world persists, especially in countries south of Saharan Africa. The efforts and concerns invested by the World Health Organization and other health agencies to eradicate malaria are commendable. However, in spite of all these efforts, the loss in economic and human resources continues. In a previous report, the long-term health effects of insecticide-impregnated bednet (IIBN) use were highlighted with the expectation of attracting serious thoughts and further research on the issue. This present paper is an update on that expectation. Results from a comprehensive literature search show that not much work has been done on the effects of long-term exposure to IIBNs in combating malarial infection. The efficacy of IIBNs is not in question. What is in question is whether long-term exposure to IIBNs have any health effects. The aims and outcomes of the research found in the literature on the subject to date seem to support only the efficacy of the temporal use of plain bednets, but not the use of IIBNs, and do not tell much about the long-term effects of IIBN exposure. All pesticides are toxic by nature and present risks of adverse effects. While there is agreement that IIBNs can be effective in reducing malarial morbidity and mortality under field trials, a number of factors relating to their long-term-exposure health effects have yet to be determined. Further reliable research projects are recommended urgently. However, some of the anticipated behavioral effects caused by insecticidal use will be avoided by the use of untreated nets instead.


Inducible DNA repair enzymes involved in the adaptive response to alkylating agents.
Two different DNA repair enzymes are induced in E. coli by methylating agents: a methyltransferase acting on 0(6)-methylguanine residues, and a DNA glycosylate for N-methylated purines. The properties of the homogeneous methyltransferase are described.


Phosphorylation of the PTEN tail regulates protein stability and function.
The PTEN gene is a tumor suppressor localized in the frequently altered chromosomal region 10q23. The tumor suppressor function of the PTEN protein (PTEN) has been linked to its ability to dephosphorylate the lipid second-messenger phosphatidylinositol 3,4, 5-trisphosphate and phosphatidylinositol 3,4-bisphosphate and, by doing so, to antagonize the phosphoinositide 3-kinase pathway. The PTEN protein consists of an amino-terminal phosphatase domain, a lipid binding C2 domain, and a 50-amino-acid C-terminal domain (the "tail") of unknown function. A number of studies have shown that the tail is dispensable for both phosphatase activity and blocking cell growth. Here, we show that the PTEN tail is necessary for maintaining protein stability and that it also acts to inhibit PTEN function. Thus, removing the tail results in a loss of stability but does not result in a loss of function because the resultant protein is more active. Furthermore, tail-dependent regulation of stability and activity is linked to the phosphorylation of three residues (S380, T382, and T383) within the tail. Therefore, the tail is likely to mediate the regulation of PTEN function through phosphorylation.


Real-time outdoor concealed-object detection with passive millimeter wave imaging.
Millimeter wave imaging is finding rapid adoption in security applications such as the detection of objects concealed under clothing. A passive imaging system can be realized as a stand-off type sensor that can operate in open spaces, both indoors and outdoors. In this paper, we address real-time outdoor concealed-object detection and segmentation with a radiometric imaging system operating in the W-band. The imaging system is equipped with a dielectric lens and a receiver array operating at around 94 GHz. Images are analyzed by multilevel segmentation to identify a concealed object. Each level of segmentation comprises vector quantization, expectation-maximization, and Bayesian decision making to cluster pixels on the basis of a Gaussian mixture model. In addition, we describe a faster process that adopts only vector quantization for the first level segmentation. Experiments confirm that the proposed methods provide fast and reliable detection and segmentation for a moving human subject carrying a concealed gun.


A study of metatarsal fractures in children.
Metatarsal fractures are common in children. The aim of the present study was to analyze a consecutive series of metatarsal fractures in children and to describe the epidemiology of the fractures, the location of the fractures, and the mechanism of injury. A consecutive series of 125 patients with metatarsal fractures who presented to one pediatric surgery department over a twenty-two-month period was evaluated with use of a questionnaire to define the mechanism of injury. All patients were followed until the fracture was healed. The specific location of the fractures and the mechanism of injury were identified. One hundred and twenty-five children (seventy-five boys and fifty girls; average age, 8.6 years [range, one to seventeen years]) presented with 166 metatarsal fractures. Major differences were found between children who were five years of age or less and those who were more than five years of age. In patients who were five years of age or less, the predominant location of the accident that had caused the fracture was inside the house and the primary mechanism was a fall from a height. In patients who were more than five years of age, most accidents occurred at sports facilities and were caused by a fall on a level surface. The most frequently fractured metatarsal in younger children was the first, whereas the most frequently fractured metatarsal in older children was the fifth. The mechanism of injury and clinical presentation of metatarsal fractures in children are age-dependent, with patients five years of age or less and those more than five years of age having different mechanisms of injury and different types of metatarsal fractures.


Turgid female toads give males the slip: a new mechanism of female mate choice in the Anura.
In many anuran species, males vocalize to attract females but will grasp any female that comes within reach and retain their hold unless displaced by a rival male. Thus, female anurans may face strong selection to repel unwanted suitors, but no mechanism is known for doing so. We suggest that a defensive trait (the ability to inflate the body to ward off attack) has been co-opted for this role: by inflating their bodies, females are more difficult for males to grasp and hence, it is easier for another male to displace an already amplexed rival. Inflating a model female cane toad (Bufo marinus) strongly reduced a male's ability to maintain amplexus; and females who were experimentally prevented from inflating their bodies experienced no successful takeovers from rival males, in contrast to control females. Thus, the ability of a female cane toad to inflate her body may allow her to manipulate the outcome of male-male competition. This overlooked mechanism of anuran mate choice may reflect a common evolutionary pattern, whereby females co-opt defensive traits for use in sexual selection.


Preference-based quality of life in patients with cerebral aneurysms.
Functional outcome scales are typically used to measure quality of life (QOL) and outcomes in patients with cerebral aneurysms; however, these instruments only examine a limited number of domains that contribute to QOL. An alternative are preference-based QOL methods, which integrate all factors contributing to QOL and provide a comprehensive individualized measure of how patients value their current health state. An additional advantage of preference-based QOL values is that they can be incorporated into decision analyses and cost-effectiveness analyses. We used 4 preference-based QOL methods to measure QOL in 176 outpatients with cerebral aneurysms: (1) standard gamble; (2) time trade-off; (3) visual analogue scale; and (4) willingness to pay. We measured functional status with the Glasgow Outcome Scale (GOS), Rankin Scale, and Barthel Index. We then built multivariate linear regression models to examine the relationships between preference-based QOL, functional status, and patient characteristics. Preference-based QOL was moderately diminished in the aneurysm patients. Mean values were: standard gamble, 0.78; time trade-off, 0.79; visual analogue scale, 0.67; and willingness to pay, 121,000 dollars. Preference-based QOL was not well-explained by functional status or patient characteristics, as shown by regression models that accounted for <15% of the variation in preference-based QOL (R2<0.15). Preference-based QOL instruments capture components of QOL in patients with cerebral aneurysms not assessed by functional status measures or patient characteristics. Studies of patients with cerebral aneurysms should consider incorporating preference-based QOL measures for a fuller evaluation of the impact of aneurysmal disease and its treatment on QOL.


Surface Chemistry of Atomically Precise Coinage-Metal Nanoclusters: From Structural Control to Surface Reactivity and Catalysis.
A comprehensive understanding of chemical bonding and reactions at the surface of nanomaterials is of great importance in the rational design of their functional properties and applications. With the rapid development in cluster science, it has become clear that atomically precise metal clusters represent ideal models for resolving various important and/or unsolved issues related to surface science. This Account highlights our recent efforts on the fabrication of ligand-stabilized coinage nanoclusters with atomic precision from the viewpoint of surface coordination chemistry in particular. The successful synthesis of a large variety of metal clusters in our group has greatly benefitted from the development of an effective amine-assisted NaBH4 reduction method. First discussed in this Account is how the introduction of amines in the synthetic protocol enhances the long-term stability and high-yield production of Ag/Cu-based metals in air. Such a method allows the utilization of different organic ligands as surface stabilizing agents to manipulate both the core and surface structures of metal nanoclusters, helping to understand the role of surface ligands in determining the structures of metal nanoclusters. The coordination chemistry of ligands used in the synthesis of metal nanoclusters is crucial in determining their overall shape, metal arrangement, surface ligand binding structure, chirality and also metal exposure. Detailed discussions are given in the following four different systems: (1) The co-use of phosphines and thiolates with rich coordination structures (2 to 4-coordinated) helps to control the formation of a sequence of Ag nanoclusters with a near-perfectly cubic shape; (2) The metal arrangements and surface structures of AuCu clusters highly depend on metal precursors and counter cations used in the synthesis; (3) Metal clusters with intrinsic chirality are readily prepared by introducing chiral ligands or counterions, making it possible to obtain optically active enantiomers and understand the origin of chirality of metal nanoclusters; (4) The variation of metal exposure of the inner metal core of metal nanocluster can be controlled by the surface ligand coordination structure. Such capabilities to manipulate the surface structure of metal nanoclusters allow the creation of model systems for investigating the structure-reactivity relationship of metal nanomaterials. Several important examples are then discussed to highlight the importance of ligand coordination chemistry in tuning the surface reactivity and catalysis of metal nanoclusters. For example, bulky thiolates on Ag are demonstrated to be more labile than small thiolates for making metal nanoclusters with both enhanced ligand exchange capability and catalysis. Alkynyl ligands can be thermally released from metal nanoclusters more easily than thiolates and halides while maintaining the overall structure, thereby serving as ideal systems for understanding the promoting effect of surface stabilizers on catalysis. Finally, we provide a perspective on the principles of surface coordination chemistry of metal nanoclusters and their potential applications with regards to catalysis of protected metal clusters.


Clinical pharmacokinetics of N-acetylcysteine.
N-Acetylcysteine is useful as a mucolytic agent for treatment of chronic bronchitis and other pulmonary diseases complicated by the production of viscous mucus. It is also used as an antidote to paracetamol (acetaminophen) poisoning and found to be effective for the prevention of cardiotoxicity by doxorubicin and haemorrhagic cystitis from oxazaphosphorines. After an oral dose of N-acetylcysteine 200 to 400 mg the peak plasma concentration of 0.35 to 4 mg/L is achieved within 1 to 2 hours. Although the data are conflicting, it appears that the administration of charcoal may interfere with drug absorption, with up to 96% of the drug adsorbed on to the charcoal. Information on absorption in the presence of food or other drugs is not available. The volume of distribution ranges from 0.33 to 0.47 L/kg and protein binding is significant, reaching approximately 50% 4 hours after the dose. Pharmacokinetic information is not available as to whether or not N-acetylcysteine crosses the blood-brain barrier or placenta, or into breast milk. Renal clearance has been reported as 0.190 to 0.211 L/h/kg and approximately 70% of the total body clearance is nonrenal. Following oral administration, reduced N-acetylcysteine has a terminal half-life of 6.25h. Little is known of the metabolism of this agent, although it is believed to be rapidly metabolised and incorporated on to proteins. The major excretory product is inorganic sulphate. Frequently reported side effects are nausea, vomiting and diarrhoea. Biochemical and haematological adverse effects are observed but are not clinically relevant. Drug interactions of clinical significance have been observed with paracetamol, glutathione and anticancer agents.


Attachment and growth of human embryonic stem cells on microcarriers.
The use of human embryonic stem cells (hESCs) for cell-based therapies will require large quantities of genetically stable pluripotent cells and their differentiated progeny. Traditional hESC propagation entails adherent culture and is sensitive to enzymatic dissociation. These constraints hamper modifying method from 2-dimensional flat-bed culture, which is expensive and impractical for bulk cell production. Large-scale culture for clinical use will require innovations such as suspension culture for bioprocessing. Here we describe the attachment and growth kinetics of both murine embryonic stem cells (mESCs) and hESCs on trimethyl ammonium-coated polystyrene microcarriers for feeder-free, 3-dimensional suspension culture. mESCs adhered and expanded according to standard growth kinetics. For hESC studies, we tested aggregate (collagenase-dissociated) and single-cell (TrypLE-dissociated) culture. Cells attached rapidly to beads followed by proliferation. Single-cell cultures expanded 3-fold over approximately 5 days, slightly exceeding that of hESC aggregates. Importantly, single-cell cultures were maintained through 6 passages with a 14-fold increase in cell number while still expressing the undifferentiated markers Oct-4 and Tra 1-81. Finally, hESCs retained their capacity to differentiate towards pancreatic, neuronal, and cardiomyocyte lineages. Our studies provide proof-of-principle of suspension-based expansion of hESCs on microcarriers, as a novel, economical and practical feeder-free means of bulk hESC production.


Fluoride-Induced Autophagy via the Regulation of Phosphorylation of Mammalian Targets of Rapamycin in Mice Leydig Cells.
Fluoride is known to impair testicular function and decrease testosterone levels, yet the underlying mechanisms remain inconclusive. The objective of this study is to investigate the roles of autophagy in fluoride-induced male reproductive toxicity using both in vivo and in vitro Leydig cell models. Using transmission electron microscopy and monodansylcadaverine staining, we observed increasing numbers of autophagosomes in testicular tissue, especially in Leydig cells of fluoride-exposed mice. Further study revealed that fluoride increased the levels of mRNA and protein expression of autophagy markers LC3, Beclin1, and Atg 5 in primary Leydig cells. Furthermore, fluoride inhibited the phosphorylation of mammalian targets of rapamycin and 4EBP1, which in turn resulted in a decrease in the levels of AKT and PI3K mRNA expression, as well as an elevation of the level of AMPK expression in both testes and primary Leydig cells. Additionally, fluoride exposure significantly changed the mRNA expression of the PDK1, TSC, and Atg13 regulator genes in primary Leydig cells but not in testicular cells. Taken together, our findings highlight the roles of autophagy in fluoride-induced testicular and Leydig cell damage and contribute to the elucidation of the underlying mechanisms of fluoride-induced male reproductive toxicity.


Odor stimuli trigger influx of calcium into olfactory neurons of the channel catfish.
Olfactory transduction is thought to be mediated by a G protein-coupled increase in intracellular adenosine 3',5'-monophosphate (cAMP) that triggers the opening of cAMP-gated cation channels and results in depolarization of the plasma membrane of olfactory neurons. In olfactory neurons isolated from the channel catfish, Ictalurus punctatus, stimulation with olfactory stimuli (amino acids) elicits an influx of calcium that leads to a rapid increase in intracellular calcium. In addition, in a reconstitution assay a plasma membrane calcium channel has been identified that is gated by inositol-1,4,5-trisphosphate (IP3), which could mediate this calcium influx. Together with previous studies indicating that stimulation with olfactory stimuli leads to stimulation of phosphoinositide turnover in olfactory cilia, these data suggest that an influx of calcium triggered by odor stimulation of phosphoinositide turnover may be an alternate or additional mechanism of olfactory transduction.


Continental arc volcanism as the principal driver of icehouse-greenhouse variability.
Variations in continental volcanic arc emissions have the potential to control atmospheric carbon dioxide (CO2) levels and climate change on multimillion-year time scales. Here we present a compilation of ~120,000 detrital zircon uranium-lead (U-Pb) ages from global sedimentary deposits as a proxy to track the spatial distribution of continental magmatic arc systems from the Cryogenian period to the present. These data demonstrate a direct relationship between global arc activity and major climate shifts: Widespread continental arcs correspond with prominent early Paleozoic and Mesozoic greenhouse climates, whereas reduced continental arc activity corresponds with icehouse climates of the Cryogenian, Late Ordovician, late Paleozoic, and Cenozoic. This persistent coupled behavior provides evidence that continental volcanic outgassing drove long-term shifts in atmospheric CO2 levels over the past ~720 million years.


Flight periodicity and the vertical distribution of high-altitude moth migration over southern Britain.
The continuous operation of insect-monitoring radars in the UK has permitted, for the first time, the characterization of various phenomena associated with high-altitude migration of large insects over this part of northern Europe. Previous studies have taken a case-study approach, concentrating on a small number of nights of particular interest. Here, combining data from two radars, and from an extensive suction- and light-trapping network, we have undertaken a more systematic, longer-term study of diel flight periodicity and vertical distribution of macro-insects in the atmosphere. Firstly, we identify general features of insect abundance and stratification, occurring during the 24-hour cycle, which emerge from four years' aggregated radar data for the summer months in southern Britain. These features include mass emigrations at dusk and, to a lesser extent, at dawn and daytime concentrations associated with thermal convection. We then focus our attention on the well-defined layers of large nocturnal migrants that form in the early evening, usually at heights of 200-500 m above ground. We present evidence from both radar and trap data that these nocturnal layers are composed mainly of noctuid moths, with species such as Noctua pronuba, Autographa gamma, Agrotis exclamationis, A. segetum, Xestia c-nigrum and Phlogophora meticulosa predominating.


Guidelines and educational outreach visits from community pharmacists to improve prescribing in general practice: a randomised controlled trial.
To evaluate the effectiveness of guidelines with or without one-to-one educational outreach visits by community pharmacists in improving general practice prescribing for non-steroidal anti-inflammatory drugs (NSAIDs). Cluster randomised trial of 20 general practices within Avon, England. Practices were randomised to three groups: control; mailed guidelines; mailed guidelines plus educational outreach visits. General practitioners (GPs) in the latter group received two one-to-one outreach visits from community pharmacists. Changes in prescribing were measured using outcomes derived from prescribing analysis and cost (PACT) data. The primary outcome measure was change in the volume of prescribing for ibuprofen, diclofenac and naproxen as a percentage of total NSAID prescribing. Six secondary outcomes included other measures of prescribing quality and volume. A cost-benefit analysis was performed. No significant differences were observed for the primary outcome measure: practices receiving outreach visits prescribed only 2.1% [95% confidence interval (CI): -0.8 to 5.0] more of the three recommended NSAIDs than the control practices did and 1.6% (95% CI: -1.4 to 4.7) more than practices that received guidelines only. Following adjustment for multiple comparisons, only one secondary outcome showed a statistically significant difference between the groups: the proportion of prescribing of the five most frequently used drugs was 2.2% (95% CI: 0.9 to 3.6) higher in the educational outreach group compared with the control group. A net increase in costs was shown with both interventions. Although good prescribing at baseline in the participating practices limited the capacity for improvement, this trial provides no evidence that guidelines with or without educational outreach visits from community pharmacists lead to substantial improvements in prescribing behaviour.


Noise assessment of a Fourier transform infrared spectroradiometer subject to the stability of a conventional laboratory blackbody source.
A method is described for the measurement of the noise-equivalent spectral radiance (NESR) of Fourier transform infrared (FTIR) spectroradiometers at all wave numbers of a selected range. The method requires minimal detailed knowledge of the sensor and no support equipment beyond a blackbody source. The NESRs of the FTIR spectroradiometer are determined at every wave-number increment in the 700-1300 cm(-1) range, for six resolutions, with a conventional blackbody source and ensembles of differential spectra. The NESRs are well behaved and consistent with the expected dependence on resolution; however, they depend on source temperature at the highest (1 cm(-1)) and lowest (32 cm(-1)) resolutions, with little or no statistical dependence at intermediate resolutions. Residual source drift is shown to be the likely cause of the dependence at 1 cm(-1); the dependence on the source at 32 cm(-1) resolution is shown to be most probably due to photon noise. At intermediate resolutions the sensor noise is dominant.


Species specificity of the Listeria monocytogenes InlB protein.
InlA and InlB mediate L. monocytogenes entry into eukaryotic cells. InlA is required for the crossing of the intestinal and placental barriers. InlA uses E-cadherin as receptor in a species-specific manner. The human E-cadherin but not the mouse E-cadherin is a receptor for InlA. In human cells, InlB uses Met and gC1qR as receptors. By studying the role of InlB in vivo, we found that activation of Met by InlB is species-specific. In mice, InlB is important for liver and spleen colonization, but not for the crossing of the intestinal epithelium. Strikingly, the virulence of a DeltainlB deletion mutant is not attenuated in guinea pigs and rabbits. Guinea pig and rabbit cell lines do not respond to InlB, although expressing Met and gC1qR, but support InlB-mediated responses upon human Met gene transfection, indicating that InlB does not recognize or stimulate guinea pig and rabbit Met. In guinea pig cells, the effect of human Met gene transfection on InlB-dependent entry is increased upon cotransfection with human gc1qr gene, showing the additive roles of gC1qR and Met. These results unravel a second L. monocytogenes species specificity critical for understanding human listeriosis and emphasize the need for developing new animal models for studying InlA and InlB functions in the same animal model.


Sanguinarine Induces Apoptosis of Human Oral Squamous Cell Carcinoma KB Cells via Inactivation of the PI3K/Akt Signaling Pathway.
Preclinical Research Sanguinarine, an alkaloid isolated from the root of Sanguinaria canadensis and other plants of the Papaveraceae family, selectively induces apoptotic cell death in a variety of human cancer cells, but its mechanism of action requires further elaboration. The present study investigated the pro-apoptotic effects of sanguinarine in human oral squamous cell carcinoma KB cells. Sanguinarine treatment increased DR5/TRAILR2 (death receptor 5/TRAIL receptor 2) expression and enhanced the activation of caspase-8 and cleavage of its substrate, Bid. Sanguinarine also induced the mitochondrial translocation of pro-apoptotic Bax, mitochondrial dysfunction, cytochrome c release to the cytosol, and activation of caspase-9 and -3. However, a pan-caspase inhibitor, z-VAD-fmk, reversed the growth inhibition and apoptosis induced by sanguinarine. Sanguinarine also suppressed the phosphorylation of phosphoinositide 3-kinase (PI3K) and Akt in KB cells, while co-treatment of cells with sanguinarine and a PI3K inhibitor revealed synergistic apoptotic effects. However, pharmacological inhibition of AMP-activated protein kinase and mitogen-activated protein kinases did not reduce or enhance sanguinarine-induced growth inhibition and apoptosis. Collectively, these findings indicate that the pro-apoptotic effects of sanguinarine in KB cells may be regulated by a caspase-dependent cascade via activation of both intrinsic and extrinsic signaling pathways and inactivation of PI3K/Akt signaling. Drug Dev Res 77 : 227-240, 2016. © 2016 Wiley Periodicals, Inc.


HPV testing in patients with low grade cervical cytological abnormalities: a follow up study.
To assess the diagnostic performance of human papillomavirus (HPV) analysis in predicting cervical intraepithelial neoplasia (CIN) grades 2 and 3 in patients with persistent low grade cervical cytological abnormalities. Cervical smears from 167 women referred for colposcopy with persistent borderline, wart virus or mildly dyskaryotic changes on cervical screening were analysed by Papanicolaou staining, non-isotopic in situ hybridisation and generic and type specific polymerase chain reaction (PCR) amplification of HPV sequences. Follow up was by cytological and, where appropriate, histological analysis. CIN grade 2 or 3 was identified in 46 patients after a median follow up of 27 months. HPV positivity by both techniques was associated with high grade CIN and with age less than 30 years (median age 33 years). Non-isotopic in situ hybridisation was more predictive but less sensitive than either generic or type specific PCR, but prediction was greater using either molecular technique in women over 30 years of age. Although the degree of prediction found is of only limited clinical value, the strong association of HPV positivity with both high grade CIN and patient age suggests that further studies of HPV testing in this patient group are warranted.


Mental health disorders after traumatic brain injury in a New Zealand caseload.
This study investigated psychological disorders for patients receiving traumatic brain injury (TBI) rehabilitation within a community-based TBI service in New Zealand. Cross-sectional study involving retrospective review of clinical notes for all referrals received in a 6-month period. This methodology was selected to gain a sample of individuals reflective of the range of patients seen in TBI rehabilitation services in New Zealand. The clinical records for 279 patients were reviewed for formal psychiatric diagnoses and references to moderate or severe psychological symptoms. Of the clinical records, 57.7% noted significant psychological problems post-injury (n = 161) and 8.6% had a psychiatric diagnosis recorded, with 10.8% reporting psychological disorders pre-injury. In comparison, 15.1% had input from a clinical psychologist and 2.2% had involvement from a mental health team. The rate of psychological symptoms was significantly higher post-TBI than pre-TBI and few of the patients reporting significant psychological symptoms were receiving intervention from mental health professionals. Further research is needed regarding the ability of TBI rehabilitation professionals in New Zealand to identify and respond to the psychological implications of TBI and on the ability of TBI and mental health teams to liaise effectively.


PTSD in prison settings: A systematic review and meta-analysis of comorbid mental disorders and problematic behaviours.
Prevalence rates of PTSD are higher in the prison population than in the community. We sought to systematically review the extent to which this disorder is associated with other mental health disorders and problematic suicidal or aggressive behaviours in the prison population. Studies reporting a relationship between PTSD and comorbid mental disorders and/or problematic behaviours in imprisoned adolescent and adult populations were identified from four bibliographic indexes. Primary studies involving clinical interviews, validated instruments leading to DSM or ICD diagnoses, or validated self-report questionnaires such as the PTSD checklist were included. Random-effects meta-analysis was conducted where possible. Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed. This review identified 36 studies, with a combined sample of 9594 participants, (6478 male and 2847 female prisoners) from 11 countries. Thirty-four of the identified studies employed a cross-sectional design. We identified significant associations between PTSD and comorbid mental disorders including depression (OR = 3.4, 95% confidence interval (CI): 2.3-4.9), anxiety (OR = 2.9, 95% confidence interval (CI): 1.8-4.7) and substance use (OR = 1.9, 95% confidence interval (CI): 1.5-2.4). We also identified significant associations between PTSD and suicidality (OR = 3, 95% confidence interval (CI): 2.4-3.8) and aggressive behaviours (this latter finding was not subject to meta-analysis). Significant methodological heterogeneity was identified between studies. High rates of psychiatric comorbidity among prisoners with PTSD, and links to suicidal behaviour, self-harm and aggressive behaviour, provide further support for the need for trauma-informed treatment approaches in prisons. However, significant gaps in the current evidence were apparent. In particular, a lack of large, longitudinal studies meant that the temporal relationships between PTSD and relevant outcomes cannot currently be determined.


Risk factors of hepatitis B transmission in northern Palestine: a case - control study.
The Hepatitis B (HB) infection is a significant health problem in Palestine, which is categorized as an HB virus moderate endemic area, with the HB carrier rate ranging from 2-6%. The aim of this study is to determine the risk factors of Hepatitis transmission in the northern areas of Palestine in order to help prevent and control this prevalent health problem. A case-control study was implemented to achieve the study objectives. One hundred HB virus seropositive cases and another 100 seronegative controls were included in the study. Univariate analysis and a logistic regression model were performed to examine probable risk factors of acquisition of HB infections. Univariate analysis showed that HB case-patients were more likely to report having a history of blood transfusion, dental visits, hospitalization, Hejamat, sharing shaving equipments, intravenous drug use, or living abroad than controls were. The logistic regression model revealed a history of dental visits to be the most significant risk factor, (P value <0.001, OR 5.6; 95% CI 2.8-11.1). The presence of these risk factors emphasizes the need for both increasing the uptake of HB vaccine and implementing risk-targeted public health education. Development and enforcement of appropriate infection control guidelines for dental care services are important to prevent HB virus transmission as well.


Understanding safety in prehospital emergency medical services for children.
For over a decade, the field of medicine has recognized the importance of studying and designing strategies to prevent safety issues in hospitals and clinics. However, there has been less focus on understanding safety in prehospital emergency medical services (EMS), particularly in regard to children. Roughly 27.7 million (or 27%) of the annual emergency department visits are by children under the age of 19, and about 2 million of these children reach the hospital via EMS. This paper adds to our qualitative understanding of the nature and contributors to safety events in the prehospital emergency care of children. We conducted four 8- to 12-person focus groups among paid and volunteer EMS providers to understand 1) patient safety issues that occur in the prehospital care of children, and 2) factors that contribute to these safety issues (e.g., patient, family, systems, environmental, or individual provider factors). Focus groups were conducted in rural and urban settings. Interview transcripts were coded for overarching themes. Key factors and themes identified in the analysis were grouped into categories using an ecological approach that distinguishes between systems, team, child and family, and individual provider level contributors. At the systems level, focus group participants cited challenges such as lack of appropriately sized equipment or standardized pediatric medication dosages, insufficient human resources, limited pediatric training and experience, and aspects of emergency medical services culture. EMS team level factors centered on communication with other EMS providers (both prehospital and hospital). Family and child factors included communication barriers and challenging clinical situations or scene characteristics. Finally, focus group participants highlighted a range of provider level factors, including heightened levels of anxiety, insufficient experience and training with children, and errors in assessment and decision making. The findings of our study suggest that, just as in hospital medicine, factors at the systems, team, child/family, and individual provider level system contribute to errors in prehospital emergency care. These factors may be modifiable through interventions and systems improvements. Future studies are needed to ascertain the generalizability of these findings and further refine the underlying mechanisms.


In Vitro and In Vivo Response to Low-Modulus PMMA-Based Bone Cement.
The high stiffness of acrylic bone cements has been hypothesized to contribute to the increased number of fractures encountered after vertebroplasty, which has led to the development of low-modulus cements. However, there is no data available on the in vivo biocompatibility of any low-modulus cement. In this study, the in vitro cytotoxicity and in vivo biocompatibility of two types of low-modulus acrylic cements, one modified with castor oil and one with linoleic acid, were evaluated using human osteoblast-like cells and a rodent model, respectively. While the in vitro cytotoxicity appeared somewhat affected by the castor oil and linoleic acid additions, no difference could be found in the in vivo response to these cements in comparison to the base, commercially available cement, in terms of histology and flow cytometry analysis of the presence of immune cells. Furthermore, the in vivo radiopacity of the cements appeared unaltered. While these results are promising, the mechanical behavior of these cements in vivo remains to be investigated.


Trans-10,cis-12-CLA-caused lipodystrophy is associated with profound changes of fatty acid profiles of liver, white adipose tissue and erythrocytes in mice: possible link to tissue-specific alterations of fatty acid desaturation.
Dietary supplementation with conjugated linoleic acid (CLA) has been shown to reduce body fat mass. To investigate the effects of individual CLA isomers on the fatty acid profiles of lipogenic (liver and white adipose) and lipid sensitive (erythrocyte) tissues, BALB/c mice were fed with 1 of 2 diets supplemented with either a c9,t11-CLA-enriched and t10,c12-CLA-free or a CLA-mixture containing both isomers in equal amounts (1% w/w of the diet) for 5 weeks. A control group was fed with a diet enriched in sunflower oil to energy balance the CLA. Compared to the t10,c12-CLA-free and the control diets, we observed a significant reduction of adipose tissue accompanied by fatty livers in the CLA-mix-fed group. These alterations in body fat distribution entailed a conspicuous shift of the fatty acid profiles of adipose tissue and livers. Liver enlargement was mainly caused by accumulation of C18 monoenes that accounted for 67 ± 1% of total fatty acid methyl esters. The significant reduction of the 18:0/18:1 desaturation index in the liver upon CLA-mix diet indicated high stearoyl-CoA desaturase activity. In contrast, reduction in white adipose tissue was largely driven by percental reduction of monounsaturated fatty acids (p ≤ 0.001). 16:0/ 16:1 and 18:0/18:1 desaturation indices for white adipose tissue significantly increased, suggesting an inhibition of stearoyl-CoA desaturase upon CLA-mix diet. The fatty acid profile of the erythrocytes widely reflected that of livers, depending on the supplemented diet. These profound changes in fatty acid composition of lipogenic organs due to t10,c12-CLA intake may be the consequence of functional alterations of lipid metabolism.


Comparison of the predictive validity of diagnosis-based risk adjusters for clinical outcomes.
Many possible methods of risk adjustment exist, but there is a dearth of comparative data on their performance. We compared the predictive validity of 2 widely used methods (Diagnostic Cost Groups [DCGs] and Adjusted Clinical Groups [ACGs]) for 2 clinical outcomes using a large national sample of patients. We studied all patients who used Veterans Health Administration (VA) medical services in fiscal year (FY) 2001 (n = 3,069,168) and assigned both a DCG and an ACG to each. We used logistic regression analyses to compare predictive ability for death or long-term care (LTC) hospitalization for age/gender models, DCG models, and ACG models. We also assessed the effect of adding age to the DCG and ACG models. Patients in the highest DCG categories, indicating higher severity of illness, were more likely to die or to require LTC hospitalization. Surprisingly, the age/gender model predicted death slightly more accurately than the ACG model (c-statistic of 0.710 versus 0.700, respectively). The addition of age to the ACG model improved the c-statistic to 0.768. The highest c-statistic for prediction of death was obtained with a DCG/age model (0.830). The lowest c-statistics were obtained for age/gender models for LTC hospitalization (c-statistic 0.593). The c-statistic for use of ACGs to predict LTC hospitalization was 0.783, and improved to 0.792 with the addition of age. The c-statistics for use of DCGs and DCG/age to predict LTC hospitalization were 0.885 and 0.890, respectively, indicating the best prediction. We found that risk adjusters based upon diagnoses predicted an increased likelihood of death or LTC hospitalization, exhibiting good predictive validity. In this comparative analysis using VA data, DCG models were generally superior to ACG models in predicting clinical outcomes, although ACG model performance was enhanced by the addition of age.


Shifts from glucose to certain secondary carbon-sources result in activation of the extracytoplasmic function sigma factor sigmaE in Salmonella enterica serovar Typhimurium.
Salmonella enterica serovar Typhimurium (S. Typhimurium) elicits the starvation-stress response (SSR) due to starvation for an essential nutrient, e.g. a carbon/energy source (C-source). As part of the SSR, the alternative sigma factor sigma(E) is activated and induced. The authors suspect that this activation is, in part, triggered by changes in the S. Typhimurium cell envelope occurring during the adaptation from growth to carbon/energy starvation (C-starvation), and resulting in an increased need for sigma(E)-regulated factors involved in the proper folding and assembly of newly synthesized proteins destined for this extracytoplasmic compartment. This led to the hypothesis that a sigma(E) activation signal might arise during C-source shifts that cause the induction of proteins localized to the extracytoplasmic compartment, i.e. the outer membrane or periplasm, of the cell. To test this hypothesis, cultures were grown in minimal medium containing enough glucose to reach mid-exponential-phase, plus a non-limiting amount of a secondary 'less-preferred' but utilizable carbon/energy source. The sigma(E) activity was then monitored using plasmids carrying rpoEP1- and rpoEP2-lacZ transcriptional fusions, which exhibit sigma(E)-independent and -dependent lacZ expression, respectively. The secondary C-sources maltose, succinate and citrate, which have extracytoplasmic components involved in their utilization (e.g. LamB), resulted in a discernible diauxic lag period and a sustained increase in sigma(E) activity. Growth transition from glucose to other utilizable phosphotransferase (PTS) and non-PTS C-sources, such as trehalose, mannose, mannitol, fructose, glycerol, d-galactose or l-arabinose, did not cause a discernible diauxic lag period or a sustained increase in sigma(E) activity. Interestingly, a shift from glucose to melibiose, which does not use an extracytoplasmic-localized protein for uptake, did cause an observable diauxic lag period but did not result in a sustained increase in sigma(E) activity. In addition, overexpression of LamB from an arabinose-inducible promoter leads to a significant increase in sigma(E) activity in the absence of a glucose to maltose shift or C-starvation. Furthermore, a DeltalamB : : Omega-Km(r) mutant, lacking the LamB maltoporin, exhibited an approximately twofold reduction in the sustained sigma(E) activity observed during a glucose to maltose shift, again supporting the hypothesis. Interestingly, the LamB protein lacks the typical Y-X-F terminal tripeptide of the OmpC-like peptides that activate DegS protease activity leading to sigma(E) activation. It does, however, possess a terminal pentapeptide (Q-M-E-I-W-W) that may function as a ligand for a putative class II PDZ-binding site. The authors therefore propose that the sigma(E) regulon of S. Typhimurium not only is induced in response to deleterious environmental conditions, but also plays a role in the adaptation of cells to new growth conditions that necessitate changes in the extracytoplasmic compartment of the cell, which may involve alternative signal recognition and activation pathways that are independent of DegS.


Physicochemical and immunocytochemical analysis of the aryl hydrocarbon receptor nuclear translocator: characterization of two monoclonal antibodies to the aryl hydrocarbon receptor nuclear translocator.
The aryl hydrocarbon receptor nuclear translocator (Arnt) is a basic helix-loop-helix transcription factor that heterodimerizes with the aryl hydrocarbon receptor to mediate signal transduction pathways inducible by 2,3,7,8-tetrachlorodibenzo-p-dioxin and other planar aromatic hydrocarbons. Monoclonal antibodies (MAbs) have been raised against a carboxyl-terminal 19-amino acid peptide hapten (MAb 2B10) and against a carboxyl-terminal 378-amino acid polypeptide-staphylococcal Protein A fusion protein (MAb 4G9) of Arnt and their characterization is described. Western blot experiments show that both MAbs specifically cross-react with an approximately 85-kDa band in cytosol prepared from COS-7 cells transfected with the full length human Arnt cDNA pBMSNeo-D24-1 and in Hepa 1c1c7 cytosol but not in Arnt-deficient Hepa 1-C4 mutant cytosol. Velocity sedimentation of Hepa 1c1c7 cytosol on sucrose gradients and Superose 6 gel permeation chromatography were used to estimate the sedimentation coefficient. Stokes radius, and relative molecular mass of Arnt as approximately 3.6-4.1 S, 6.8 nm, and 101-115 kDa, respectively. These results indicate that Arnt probably exists in monomeric form in Hepa 1c1c7 cytosolic extracts. Laser scanning confocal microscopy and indirect immunofluorescence microscopy revealed Arnt to be distributed throughout the non-nucleolar portion of the nucleus of Hepa 1c1c7, VT(2) (Hepa 1-C4T mutant cell line deficient in Arnt function and stably transfected with pBMSNeo D24-1, expressing the full length human Arnt cDNA), and HeLa cells. The establishment of the nuclear localization of Arnt in human and murine cell lines shown here indicates that its nuclear localization may be conserved across species. Immunofluorescence analysis of Arnt in three cell lines using two MAbs (to distinct epitopes) provides evidence that suggests that the aryl hydrocarbon receptor heterodimerizes with Arnt in the nucleus.


Pharmacological characterization of the cannabinoid receptor 2 agonist, β-caryophyllene on seizure models in mice.
Activation of CB1 receptors, produces anticonvulsant effect accompanied by memory disturbance both in animal seizure tests and in patients with epilepsy. Few reports considered the role of CB2 receptor on seizure susceptibility and cognitive functions. The aim of the present study was to explore the effect of a selective CB2 receptor agonist β-caryophyllene (BCP) in models of seizures and cognition in mice. Dose-dependent effects of BCP was studied in maximal electroshock seizure (MES) test, subcutaneous pentylenetetrazole (scPTZ) test and Morris water maze test. Phenytoin and diazepam were used as reference drugs in seizure tests. The effect of sub-chronic treatment with BCP for 7 days (50 and 100 mg kg-1) was assessed on status epilepticus (SE) induced by kainic acid (KA) model and oxidative stress through measurement of malondialdehyde (MDA) level in the hippocampus. The acute neurotoxicity was determined by a rotarod test. The BCP exerted a protection in the MES test at the lowest dose of 30 mg kg-1 at the 4-h interval tested comparable to that of the referent drug phenytoin. The CB2 agonist was ineffective in the scPTZ test. The BCP displayed no neurotoxicity in the rotarod test. The BCP decreased the seizure scores in the KA-induced SE, which effect correlated with a diminished lipid peroxidation. The CB2 agonist exerted a dose-dependent decrease of latency to cross the target area during the three days of testing in the Morris water maze test. Our results suggest that the CB2 receptor agonists might be clinically useful as an adjunct treatment against seizure spread and status epilepticus and concomitant oxidative stress, neurotoxicity and cognitive impairments.


When imagining yourself in pain, visual perspective matters: the neural and behavioral correlates of simulated sensory experiences.
Via mental simulation, imagined events faithfully reproduce the neural and behavioral activities that accompany their actual occurrence. However, little is known about how fundamental characteristics of mental imagery-notably perspectives of self-shape neurocognitive processes. To address this issue, we used fMRI to explore the impact that vantage point exerts on the neural and behavioral correlates of imaginary sensory experiences (i.e., pain). Participants imagined painful scenarios from three distinct visual perspectives: first-person self (1PS), third-person self (3PS), and third-person other (3PO). Corroborating increased ratings of pain and embodiment, 1PS (cf. 3PS) simulations elicited greater activity in the right anterior insula, a brain area that supports interoceptive and emotional awareness. Additionally, 1PS simulations evoked greater activity in brain areas associated with visual imagery and the sense of body ownership. Interestingly, no differences were observed between 3PS and 3PO imagery. Taken together, these findings reveal the neural and behavioral correlates of visual perspective during mental simulation.


A phase II trial of a combination of pemetrexed and gemcitabine in patients with metastatic breast cancer: an NCCTG study.
This phase II study was undertaken to define the efficacy and toxicity of pemetrexed in combination with gemcitabine in patients with metastatic breast cancer. Patients with measurable metastatic breast cancer who had previously received an anthracycline and a taxane in either the adjuvant or metastatic setting were treated with gemcitabine 1250 mg/m2 (intravenous; days 1 and 8) and pemetrexed 500 mg/m2 (intravenous; day 8) every 21 days. Fifty-nine patients received a median of five cycles (range one to 22) of treatment and were followed until death or for a median of 28 months (range 19.4-36.6) among living patients. Fourteen partial responses for an overall response rate of 24% [95% confidence interval (CI) 16% to 39%] were documented. Nine (15%; CI 5% to 32%) patients had stable disease for >6 months. The median survival time was 10.3 months (95% CI 8.3-18.9) and the 1 year survival rate was 49% (95% CI 38% to 64%). The median time to progression was estimated to be 3.7 months (95% CI 2.3-5.3). The most common grade 3 or 4 toxicities were neutropenia and thrombocytopenia in 83% and 27% of patients, respectively. Fourteen percent of patients experienced febrile neutropenia. Other common grade 3 or 4 non-hematological toxicities included fatigue (17%), dyspnea (15%), rash (7%) and anorexia (5%). The combination of pemetrexed and gemcitabine is clinically active, with an overall response rate of 24% in patients with metastatic breast cancer who have previously been treated with an anthracycline and a taxane. Myelosuppression (66% grade 4 neutropenia and 14% febrile neutropenia) was the major treatment-related toxicity observed for this combination.


Effect of long-chain ω-3 fatty acids and lutein + zeaxanthin supplements on cardiovascular outcomes: results of the Age-Related Eye Disease Study 2 (AREDS2) randomized clinical trial.
Dietary supplements have been proposed as a mechanism to improve health and prevent disease. To determine if supplementing diet with long-chain ω-3 polyunsaturated fatty acids or with macular xanthophylls results in a reduced rate of cardiovascular disease (CVD). The Cardiovascular Outcome Study (COS) was an ancillary study of the Age-Related Eye Disease Study 2 (AREDS2), a factorial-designed randomized clinical trial of 4203 participants recruited from 82 US academic and community ophthalmology clinics, who were followed up for a median of 4.8 years. Individuals were eligible to participate if they were between the ages of 50 and 85 years, had intermediate or advanced age-related macular degeneration in 1 eye, and were willing to be randomized. Participants with stable, existing CVD (>12 months since initial event) were eligible to participate. Participants, staff, and outcome assessors were masked to intervention. Daily supplementation with long-chain ω-3 polyunsaturated fatty acids (350-mg docosahexaenoic acid [DHA] + 650-mg eicosapentaenoic acid [EPA]), macular xanthophylls (10-mg lutein + 2-mg zeaxanthin), combination of the two, or matching placebos. These treatments were added to background therapy of the AREDS vitamin and mineral formulation for macular degeneration. MAIN OUTCOMES AND MEASURES A composite outcome of myocardial infarction, stroke, and cardiovascular death with 4 prespecified secondary combinations of the primary outcome with hospitalized heart failure, revascularization, or unstable angina. Study participants were primarily white, married, and highly educated, with a median age at baseline of 74 years. A total of 602 cardiovascular events were adjudicated, and 459 were found to meet 1 of the study definitions for a CVD outcome. In intention-to-treat analysis, no reduction in the risk of CVD or secondary CVD outcomes was seen for the DHA + EPA (primary outcome: hazard ratio [HR], 0.95; 95% CI, 0.78-1.17) or lutein + zeaxanthin (primary outcome: HR, 0.94; 95% CI, 0.77-1.15) groups. No differences in adverse events or serious adverse event were seen by treatment group. The sample size was sufficient to detect a 25% reduction in CVD events with 80% power. Dietary supplementation of long-chain ω-3 polyunsaturated fatty acids or macular xanthophylls in addition to daily intake of minerals and vitamins did not reduce the risk of CVD in elderly participants with age-related macular degeneration. clinicaltrials.gov Identifier: NCT00345176.


A specific exercise program for patients with subacromial impingement syndrome can improve function and reduce the need for surgery.
Does a specific exercise program improve shoulder function more than non-specific exercises in patients with subacromial impingement? Randomised, controlled trial with concealed allocation and blinded outcome assessment. University hospital in Sweden. Patients aged 30 to 65 years with subacromial impingement syndrome of at least 6 months duration, and on the waiting listing for surgery were included. Key exclusion criteria included previous shoulder fractures, and frozen shoulder. Randomisation of 102 participants allocated 52 to the intervention exercise group and 50 to a control exercise group. Both groups received a subacromial corticosteroid injection at inclusion and commenced exercises 2 weeks later. Both groups visited a physiotherapist 7 times over 10 weeks and were prescribed home exercises for 12 weeks. The intervention exercise group were prescribed 6 exercises: 2 eccentric exercises for the rotator cuff, 3 concentric/ eccentric exercises for the scapula stabilisers, and a posterior shoulder stretch. Each strengthening exercise was repeated 15 times in 3 sets twice daily for 8 weeks and then once daily for 4 weeks. The stretch was completed for 30 to 60 seconds and repeated 3 times twice daily. Training load was progressed using weights or elasticised bands. The control group exercise program consisted of 6 non-specific movement exercises for the neck and shoulder (e.g. neck retraction, shoulder abduction). The control group exercises were not loaded or progressed and were completed 10 times 3 times daily. The primary outcome was the Constant shoulder score at 3 months. The Constant score is scored from 0 to 100 with a higher score indicating better function. Secondary outcome measures included the disability of the arm, shoulder and hand questionnaire (DASH), a visual analogue score for pain, the EuroQol quality of life instrument, and whether the participant thought they still needed surgery. 97 participants completed the study. At 3 months, the change in Constant score was significantly more in the specific exercise group than the control group by 15 (95% CI 8.5 to 20.6) points. The DASH improved significantly more in the intervention than the control group by 8 (95% CI 2.3 to 13.7) points. The intervention group also improved significantly more than the control group in ratings of night pain, and quality of life. A lower proportion of the specific exercise group subsequently chose surgery (20% v 63%, Number Needed to Treat 3, 95% CI 1.6 to 3.9). A specific, progressive exercise program focusing on training the rotator cuff and scapular stabilisers was effective in improving function, reducing pain and reducing the need of surgery for patients with chronic subacromial impingement syndrome. [Numbers needed to treat and 95% CIs calculated by the CAP Editor.].


Effect of diet on type 2 diabetes mellitus: A review.
Globally, type 2 diabetes mellitus (T2DM) is considered as one of the most common diseases. The etiology of T2DM is complex and is associated with irreversible risk factors such as age, genetic, race, and ethnicity and reversible factors such as diet, physical activity and smoking. The objectives of this review are to examine various studies to explore relationship of T2DM with different dietary habits/patterns and practices and its complications. Dietary habits and sedentary lifestyle are the major factors for rapidly rising incidence of DM among developing countries. In type 2 diabetics, recently, elevated HbA1c level has also been considered as one of the leading risk factors for developing microvascular and macrovascular complications. Improvement in the elevated HbA1c level can be achieved through diet management; thus, the patients could be prevented from developing the diabetes complications. Awareness about diabetes complications and consequent improvement in dietary knowledge, attitude, and practices lead to better control of the disease. The stakeholders (health-care providers, health facilities, agencies involved in diabetes care, etc.) should encourage patients to understand the importance of diet which may help in disease management, appropriate self-care and better quality of life.


Use of beta-cyclodextrin in the capillary zone electrophoretic separation of the components of clandestine heroin preparations.
The present paper describes the methodological optimization and validation of a capillary zone electrophoresis method for the rapid determination of heroin, secondary products and additives present in clandestine heroin samples, by using 20 mM beta-cyclodextrins in phosphate buffer, pH 3.23. Applied potential was 15 kV and separation temperature was 24 degrees C; detection was by UV absorption at 200 nm wavelength. Heroin samples were first dissolved in CHCl3-MeOH (96:4, v/v) and injected by pressure (0.5 p.s.i., 3 s; 1 p.s.i.=6894.76 Pa) after evaporation of the organic mixture and reconstitution in aqueous buffer. Under the described conditions, phenylethylamine (internal standard), morphine, monoacetylmorphine, heroin, acetylcodeine, papaverine, codeine and narcotine were baseline resolved in less than 10 min. The limit of detection was better than 1 microg/ml for each analyte. The study of the intra-day and day-to-day precision showed, in terms of migration times, RSDs < or = 0.71% and, in terms of peak areas, RSDs < or = 3.2%. Also, the evaluation of linearity and analytical accuracy of the method provided good results for all the analytes investigated, thus allowing its application to real cases of seized controlled drug preparations.


Thapsigargin modulates agonist-stimulated cyclic AMP responses through cytosolic calcium-dependent and -independent mechanisms in rat pinealocytes.
The role of mobilization of intracellular Ca2+ in the adrenergic-stimulated cAMP accumulation in rat pinealocytes was investigated with thapsigargin, an agent that inhibits endoplasmic reticulum Ca2+-ATPase. It was found that although thapsigargin alone had no effect on the basal cAMP accumulation, it potentiated the beta-adrenergic-stimulated cAMP response by isoproterenol in a dose-dependent manner. The potentiation was abolished with ethylene glycol bis(beta-aminoethyl ether)-N, N,N',N'-tetraacetic acid-acetoxymethyl ester (EGTA-AM) but persisted in the presence of isobutylmethylxanthine, indicating that thapsigargin enhances cAMP synthesis through elevation of cytosolic intracellular Ca2+ concentration ([Ca2+]i). However, when the pinealocytes were stimulated by norepinephrine, a mixed alpha 1- and beta-adrenergic agonist, thapsigargin dose-dependently inhibited the cAMP response. To investigate this inhibitory effect of thapsigargin, we substituted ionomycin, a [Ca2+]i-elevating agent, and 4 beta-phorbol-12-myristate 13-acetate, an activator of protein kinase C, for the alpha(1)-adrenergic component of the norepinephrine-stimulated response. Although thapsigargin had no effect on the potentiation of the isoproterenol-stimulated cAMP accumulation by ionomycin, it significantly inhibited the potentiation by 4 beta-phorbol-12-myristate 13-acetate. Furthermore, the inhibitory effect of thapsigargin was not affected by cotreatment with EGTA-AM or ionomycin, suggesting that this effect is independent of [Ca2+]i. Similar results were obtained when cyclopiazonic acid was used to inhibit the Ca(2+)-ATPase. Taken together, our results indicate that thapsigargin enhances the beta-adrenergic-stimulated cAMP accumulation through its action in elevating [Ca2+]i but inhibits the potentiation of the beta-adrenergic-stimulated cAMP response by protein kinase C, as a consequence of Ca(2+)-ATPase inhibition.


Later stage at diagnosis and worse survival in cutaneous malignant melanoma among men living alone: a nationwide population-based study from Sweden.
To investigate the association between cohabitation status, clinical stage at diagnosis, and disease-specific survival in cutaneous malignant melanoma (CMM). This nationwide population-based study included 27,235 patients from the Swedish Melanoma Register diagnosed with a primary invasive CMM between 1990 and 2007 and linked data to nationwide, population-based registers followed up through 2012. After adjustment for age at diagnosis, level of education, living area, period of diagnosis, and tumor site, the odds ratios (ORs) of higher stage at diagnosis were significantly increased among men living alone versus men living with a partner (stage II v stage I: OR, 1.42; 95% CI, 1.29 to 1.57; stage III or IV v stage I: OR, 1.43; 95% CI, 1.14 to 1.79). The OR for stage II versus stage I disease was also increased among women living alone (OR, 1.15; 95% CI, 1.04 to 1.28). After adjustments for the factors listed earlier, the CMM-specific survival was significantly decreased among men living alone (hazard ratio [HR] for death, 1.48; 95% CI, 1.33 to 1.65; P < .001). After additional adjustments for all potential and established prognostic factors, CMM-specific survival among men living alone versus men living with a partner remained significantly decreased (HR, 1.31; 95% CI, 1.18 to 1.46; P < .001), suggesting a residual adverse effect on survival not accounted for by these parameters. In all age groups among men, living alone is significantly associated with reduced CMM-specific survival, partially attributed to a more advanced stage at diagnosis. This emphasizes the need for improved prevention and early detection strategies for this group.


Neurogenin2 directs granule neuroblast production and amplification while NeuroD1 specifies neuronal fate during hippocampal neurogenesis.
The specification and differentiation of dentate gyrus granule neurons in the hippocampus require temporally and spatially coordinated actions of both intrinsic and extrinsic molecules. The basic helix-loop-helix transcription factor Neurogenin2 (Ngn2) and NeuroD1 are key regulators in these processes. Based on existing classification, we analyzed the molecular events occurring during hippocampal neurogenesis, primarily focusing on juvenile animals. We found that Ngn2 is transiently expressed by late type-2a amplifying progenitors. The Ngn2 progenies mature into hippocampal granule neurons. Interestingly, the loss of Ngn2 at early stages of development leads to a robust reduction in neurogenesis, but does not disturb granule neuron maturation per se. We found that the role of Ngn2 is to maintain progenitors in an undifferentiated state, allowing them to amplify prior to their maturation into granule neurons upon NeuroD1 induction. When we overexpressed Ngn2 and NeuroD1 in vivo, we found NeuroD1 to exhibit a more pronounced neuron-inductive effect, leading to granule neuron commitment, than that displayed by Ngn2. Finally, we observed that all markers expressed during the transcriptional control of hippocampal neurogenesis in rodents are also present in the human hippocampus. Taken together, we demonstrate a critical role of for Ngn2 and NeuroD1 in controlling neuronal commitment and hippocampal granule neuroblast formation, both during embryonic development and in post-natal hippocampal granule neurogenesis.


Sea-level rise and its impact on coastal zones.
Global sea levels have risen through the 20th century. These rises will almost certainly accelerate through the 21st century and beyond because of global warming, but their magnitude remains uncertain. Key uncertainties include the possible role of the Greenland and West Antarctic ice sheets and the amplitude of regional changes in sea level. In many areas, nonclimatic components of relative sea-level change (mainly subsidence) can also be locally appreciable. Although the impacts of sea-level rise are potentially large, the application and success of adaptation are large uncertainties that require more assessment and consideration.


Jasmonate biochemical pathway.
Plants possess a family of potent fatty acid-derived wound-response and developmental regulators: the jasmonates. These compounds are derived from the tri-unsaturated fatty acids alpha-linolenic acid (18:3) and, in plants such as Arabidopsis thaliana and tomato, 7(Z)-, 10(Z)-, and 13(Z)-hexadecatrienoic acid (16:3). The lipoxygenase-catalyzed addition of molecular oxygen to alpha-linolenic acid initiates jasmonate synthesis by providing a 13-hydroperoxide substrate for formation of an unstable allene oxide by allene oxide synthase (AOS). This allene oxide then undergoes enzyme-guided cyclization to produce 12-oxophytodienoic acid (OPDA). These first steps take place in plastids, but further OPDA metabolism occurs in peroxisomes. OPDA has several fates, including esterification into plastid lipids and transformation into the 12-carbon prohormone jasmonic acid (JA). JA is itself a substrate for further diverse modifications, including the production of jasmonoyl-isoleucine (JA-Ile), which is a major biologically active jasmonate among a growing number of jasmonate derivatives. Each new jasmonate family member that is discovered provides another key to understanding the fine control of gene expression in immune responses; in the initiation and maintenance of long-distance signal transfer in response to wounding; in the regulation of fertility; and in the turnover, inactivation, and sequestration of jasmonates, among other processes.


pH-Induced Regulation of Excitation Energy Transfer in the Cyanobacterial Photosystem I Tetramer.
Excitation energy-transfer processes in pigment-protein complexes in photosynthetic organisms are often changed under different pH conditions. However, it is unclear how the pH changes affect excitation energy relaxations in photosystem I (PSI) cores. In this study, we examined the pH sensitivity of energy dynamics in the PSI tetramer, dimer, and monomer isolated from a cyanobacterium, Anabaena sp. PCC 7120, by means of time-resolved fluorescence spectroscopy. Each PSI was adapted to pH 5.0, 6.5, and 8.0. Fluorescence decay-associated (FDA) spectra of the pH 8.0 PSI dimer and monomer showed positive and negative peaks within 5 ps, whereas the FDA spectra of the PSI tetramer did not show such a 5 ps fluorescence component. Mean lifetimes of the fluorescence at pH 6.5 are shorter in the PSI tetramer than in the PSI dimer and monomer, indicating an accelerated energy quenching in the tetramer. The effects of the acidic and basic pHs on the energy-transfer processes differ significantly among the three types of PSIs, suggesting different pH-sensing sites around pigment molecules in the three PSIs. Based on these results, together with our recent structural finding of the PSI tetramer, we discuss functional implications for the pH-sensing regulation of the excitation energy transfer in the PSI tetramer.


Disparities in premature mortality between high- and low-income US counties.
Several well-established determinants of health are associated with premature mortality. Using data from the 2010 County Health Rankings, we describe the association of selected determinants of health with premature mortality among counties with broadly differing levels of income. County-level data on 3,139 US counties from the 2010 County Health Rankings were linked to county mortality data from the Centers for Disease Control and Prevention Compressed Mortality database. We divided counties into 3 groups, defined by sample median household income levels: low-income (≤25th percentile, $29,631), mid-income (25th-75th percentile, $29,631-$39,401), and high-income (≥75th percentile, ≥$39,401). We analyzed group differences in geographic, sociodemographic, racial/ethnic, health care, social, and behavioral factors. Stratified multivariable linear regression explored the associations of these health determinants with premature mortality for high- and low-income groups. The association between income and premature mortality was stronger among low-income counties than high-income counties. We found differences in the pattern of risk factors between high- and low-income groups. Significant geographic, sociodemographic, racial/ethnic, health care, social, and behavioral disparities exist among income groups. Geographic location and the percentages of adult smokers and adults with a college education were associated with premature mortality rates in US counties. These relationships varied in magnitude and significance across income groups. Our findings suggest that population health policies aimed at reducing mortality disparities require an understanding of the socioeconomic context within which modifiable variables exist.


The evolutionary history of Shigella and enteroinvasive Escherichia coli revised.
In Shigella and enteroinvasive Escherichia coli (EIEC), the etiologic agents of shigellosis in humans, the determinants responsible for entry of bacteria into and dissemination within epithelial cells are encoded by a virulence plasmid. To understand the evolution of the association between the virulence plasmid and the chromosome, we performed a phylogenetic analysis using the sequences of four chromosomal genes (trpA, trpB, pabB, and putP) and three virulence plasmid genes (ipaB, ipaD, and icsA) of a collection of 51 Shigella and EIEC strains. The phylogenetic tree derived from chromosomal genes showed a typical "star" phylogeny, indicating a fast diversification of Shigella and EIEC groups. Phylogenetic groups obtained from the chromosomal and plasmidic genes were similar, suggesting that the virulence plasmid and the chromosome share similar evolutionary histories. The few incongruences between the trees could be attributed to exchanges of fragments of different plasmids and not to the transfer of an entire plasmid. This indicates that the virulence plasmid was not transferred between the different Shigella and EIEC groups. These data support a model of evolution in which the acquisition of the virulence plasmid in an ancestral E. coli strain preceded the diversification by radiation of all Shigella and EIEC groups, which led to highly diversified but highly specialized pathogenic groups.


Clinical features and obstetric and neonatal outcomes of pregnant patients with COVID-19 in Wuhan, China: a retrospective, single-centre, descriptive study.
In December, 2019, coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, China. The number of affected pregnant women is increasing, but scarce information is available about the clinical features of COVID-19 in pregnancy. This study aimed to clarify the clinical features and obstetric and neonatal outcomes of pregnant patients with COVID-19. In this retrospective, single-centre study, we included all pregnant women with COVID-19 who were admitted to Tongji Hospital in Wuhan, China. Clinical features, treatments, and maternal and fetal outcomes were assessed. Seven patients, admitted to Tongji Hospital from Jan 1, to Feb 8, 2020, were included in our study. The mean age of the patients was 32 years (range 29-34 years) and the mean gestational age was 39 weeks plus 1 day (range 37 weeks to 41 weeks plus 2 days). Clinical manifestations were fever (six [86%] patients), cough (one [14%] patient), shortness of breath (one [14%] patient), and diarrhoea (one [14%] patient). All the patients had caesarean section within 3 days of clinical presentation with an average gestational age of 39 weeks plus 2 days. The final date of follow-up was Feb 12, 2020. The outcomes of the pregnant women and neonates were good. Three neonates were tested for SARS-CoV-2 and one neonate was infected with SARS-CoV-2 36 h after birth. The maternal, fetal, and neonatal outcomes of patients who were infected in late pregnancy appeared very good, and these outcomes were achieved with intensive, active management that might be the best practice in the absence of more robust data. The clinical characteristics of these patients with COVID-19 during pregnancy were similar to those of non-pregnant adults with COVID-19 that have been reported in the literature. National Natural Science Foundation of China, Hubei Provincial Natural Science Foundation of China.


Role of the triplet state in the green emission peak of polyfluorene films: A time evolution study.
The blue emission of ethyl-hexyl substituted polyfluorene (PF2/6) films is accompanied by a low energy green emission peak around 500 nm in inert atmosphere. The intensity of this 500 nm peak is large in electroluminescence (EL) compared to photoluminescence (PL) measurements. Furthermore, the green emission intensity reduces dramatically in the presence of molecular oxygen. To understand this, we have modeled various nonradiative processes by time dependent quantum many body methods. These are (i) intersystem crossing to study conversion of excited singlets to triplets leading to a phosphorescence emission, (ii) electron-hole recombination (e-hR) process in the presence of a paramagnetic impurity to follow the yield of triplets in a polyene system doped with paramagnetic metal atom, and (iii) quenching of excited triplet states in the presence of oxygen molecules to understand the low intensity of EL emission in ambient atmosphere, when compared with that in nitrogen atmosphere. We have employed the Pariser-Parr-Pople Hamiltonian to model the molecules and have invoked electron-electron repulsions beyond zero differential approximation while treating interactions between the organic molecule and the rest of the system. Our time evolution methods show that there is a large cross section for triplet formation in the e-hR process in the presence of paramagnetic impurity with degenerate orbitals. The triplet yield through e-hR process far exceeds that in the intersystem crossing pathway, clearly pointing to the large intensity of the 500 nm peak in EL compared to PL measurements. We have also modeled the triplet quenching process by a paramagnetic oxygen molecule which shows a sizable quenching cross section especially for systems with large sizes. These studies show that the most probable origin of the experimentally observed low energy EL emission is the triplets.


Real-time electrochemical LAMP: a rational comparative study of different DNA intercalating and non-intercalating redox probes.
We present a comparative study of ten redox-active probes for use in real-time electrochemical loop-mediated isothermal amplification (LAMP). Our main objectives were to establish the criteria that need to be fulfilled for minimizing some of the current limitations of the technique and to provide future guidelines in the search for ideal redox reporters. To ensure a reliable comparative study, each redox probe was tested under similar conditions using the same LAMP reaction and the same entirely automatized custom-made real-time electrochemical device (designed for electrochemically monitoring in real-time and in parallel up to 48 LAMP samples). Electrochemical melt curve analyses were recorded immediately at the end of each LAMP reaction. Our results show that there are a number of intercalating and non-intercalating redox compounds suitable for real-time electrochemical LAMP and that the best candidates are those able to intercalate strongly into ds-DNA but not too much to avoid inhibition of the LAMP reaction. The strongest intercalating redox probes were finally shown to provide higher LAMP sensitivity, speed, greater signal amplitude, and cleaner-cut DNA melting curves than the non-intercalating molecules.


Autoantibodies to BCOADC-E2 in patients with primary biliary cirrhosis recognize a conformational epitope.
Primary biliary cirrhosis (PBC) is an autoimmune disease of liver associated with a unique serologic response to mitochondrial autoantigens. Many of the autoantigens recognized by autoantibodies in PBC are members of the 2-oxo-acid dehydrogenase complex. The two major autoantigens are the E2 component of the pyruvate dehydrogenase complex (PDC-E2) and the E2 component of the branched chain 2-oxo-acid dehydrogenase complex (BCOADC-E2). The autoantibody response to PDC-E2 has been mapped to one immunodominant epitope, which consists of both linear and conformational components. The presence of a single immunodominant epitope in PDC-E2 is unusual when contrasted to the immune response to autoantigens in other human autoimmune diseases. We have mapped the epitope recognized by antimitochondrial autoantibodies (AMA) specific to BCOADC-E2 in patients with PBC by taking advantage of the full-length bovine BCOADC-E2 complementary DNA (cDNA) and a series of expression clones spanning the entire molecule. Reactivity to the various expression clones was studied by immunoblotting, enzyme-linked immunosorbent assay (ELISA), as well as selective absorption of patient sera by expressed protein fragments. Autoantibodies to BCOADC-E2 map within peptides spanning amino acid residues 1 to 227 of the mature protein; our data demonstrate that the epitope is dependent on conformation and includes the lipoic acid binding region. However, only the full-length clone (amino acid residue 1 to 421) is sufficient to remove all detectable BCOADC-E2 reactivity. Moreover, the absence of lipoic acid on the recombinant polypeptides used in this study indicates that antibody binding to BCOADC-E2 is not dependent on the presence of lipoic acid.


Activated carbon and biochar amendments decrease pore-water concentrations of polycyclic aromatic hydrocarbons (PAHs) in sewage sludge.
The aim of the research was to determine the influence of biochar and activated carbon (AC) on the freely dissolved concentration of polycyclic aromatic hydrocarbons (PAHs) in sewage sludge. Two different biochars (MSB and PMW) and two ACs (CP1 and BP2) were used in the present experiment. Addition of AC/biochar to sewage sludge caused significant decrease of freely dissolved PAHs concentration. Depending on the dose, the reduction of freely dissolved PAHs ranged from 56% to 95% (ACs) and from 0% to 57% (biochars). Only for the biochars was there a significant difference between short 7-d and long 30/60-d mixing time. It is concluded that both AC and biochar are effective at reducing PAH pore-water concentrations, the more expensive and non-carbon negative AC having the greatest effect.


A FancD2-monoubiquitin fusion reveals hidden functions of Fanconi anemia core complex in DNA repair.
In DNA damage responses, the Fanconi anemia (FA) protein, FancD2, is targeted to chromatin and forms nuclear foci following its monoubiquitination, a process likely catalyzed by the FA core complex. Here, we show that a chicken FancD2-ubiquitin fusion protein, carrying a Lys-Arg substitution removing the natural monoubiquitination site (D2KR-Ub), could reverse cisplatin hypersensitivity and localize to chromatin in FANCD2-deficient DT40 cells. Importantly, the chromatin targeting was dependent on three core complex components as well as the hydrophobic surface of ubiquitin that may direct protein-protein interactions. Furthermore, a constitutively chromatin bound fusion of D2KR-histone H2B could complement cisplatin sensitivity in FANCD2- but not FANCC-, FANCG-, or FANCL-deficient cells. Thus these core complex components have an additional function in the DNA repair, which is independent of the monoubiquitination and chromatin targeting of FancD2. These results define functional consequences of FancD2 monoubiquitination and reveal previously hidden functions for the FA protein core complex.


Genomic structures of viral agents in relation to the biosynthesis of selenoproteins.
The genomes of both bacteria and eukaryotic organisms are known to encode selenoproteins, using the UGA codon for seleno-cysteine (SeC), and a complex cotranslational mechanism for SeC incorporation into polypeptide chains, involving RNA stem-loop structures. These common features and similar codon usage strongly suggest that this is an ancient evolutionary development. However, the possibility that some viruses might also encode selenoproteins remained unexplored until recently. Based on an analysis of the genomic structure of the human immunodeficiency virus HIV-1, we demonstrated that several regions overlapping known HIV genes have the potential to encode selenoproteins (Taylor et al. [31], J. Med. Chem. 37, 2637-2654 [1994]). This is provocative in the light of overwhelming evidence of a role for oxidative stress in AIDS pathogenesis, and the fact that a number of viral diseases have been linked to selenium (Se) deficiency, either in humans or by in vitro and animal studies. These include HIV-AIDS, hepatitis B linked to liver disease and cancer, Coxsackie virus B3, Keshan disease, and the mouse mammary tumor virus (MMTV), against which Se is a potent chemoprotective agent. There are also established biochemical mechanisms whereby extreme Se deficiency can induce a proclotting or hemorrhagic effect, suggesting that hemorrhagic fever viruses should also be examined for potential virally encoded selenoproteins. In addition to the RNA stem-loop structures required for SeC insertion at UGA codons, genomic structural features that may be required for selenoprotein synthesis can also include ribosomal frameshift sites and RNA pseudoknots if the potential selenoprotein module overlaps with another gene, which may prove to be the rule rather than the exception in viruses. One such pseudoknot that we predicted in HIV-1 has now been verified experimentally; a similar structure can be demonstrated in precisely the same location in the reverse transcriptase coding region of hepatitis B virus. Significant new findings reported here include the existence of highly distinctive glutathione peroxidase (GSH-Px)-related sequences in Coxsackie B viruses, new theoretical data related to a previously proposed potential selenoprotein gene overlapping the HIV protease coding region, and further evidence in support of a novel frameshift site in the HIV nef gene associated with a well-conserved UGA codon in the 1-reading frame.


Epilepsy is ubiquitous, but more devastating in the poorer regions of the world... or is it?
The incidence and prevalence of active epilepsy are greatest in Africa compared to all other continents, even those with equivalent poor settings. This is a reflection of the high levels of structural and metabolic causes and may reflect an increased risk in parts of the continent. The full burden of epilepsy, which includes the social and medical morbidity of the disorder and where people with epilepsy are heavily stigmatized and frequently untreated, cannot be fully assessed even using the disability adjusted life-years, since the assigned disability weights are not specific to these regions. The burden is further exacerbated by social, geographic, and economic barriers to care and the inability of African health systems to manage people with epilepsy effectively because of lack of trained personnel, limited facilities, and poor access to effective or sustained supplies of antiepileptic drugs, or even therapy at all. The situation is compounded by a probable underestimation of the prevalence and incidence of people with epilepsy related to the major stigma associated with the condition in Africa, and the limited training available to most health care workers who are the primary point of assessing most people with epilepsy. Finding innovative ways to address the huge barriers faced by people with epilepsy in Africa needs to be a major goal for the millennium. A PowerPoint slide summarizing this article is available for download in the Supporting Information section here.


IL-18 protects mice against pulmonary and disseminated infection with Cryptococcus neoformans by inducing IFN-gamma production.
We examined the effects of a newly identified cytokine, IL-18, originally designated as IFN-gamma-inducing factor, in a mouse model of pulmonary and disseminated infection with a highly virulent strain of Cryptococcus neoformans. Administration of murine rIL-18 enhanced elimination of live micro-organisms from the lungs, prevented fungal dissemination to the brain, reduced the level of serum cryptococcal capsular polysaccharide Ags, and increased the survival rate of infected mice. Histologic examination of lung sections of infected and PBS-treated mice showed a poor cellular inflammatory reaction and a large number of multiplying C. neoformans yeast cells in alveolar spaces. In contrast, massive infiltration of inflammatory cells, consisting mainly of mononuclear cells, characterized sections of lungs of infected animals treated with IL-18. Treatment with IL-18 also increased the level of serum IFN-gamma. In addition, the protective effect of IL-18 on cryptococcal infection was abrogated by administration of neutralizing anti-IFN-gamma mAb. Finally, we examined the effect of neutralizing anti-IL-18 Ab on cryptococcal infection to define the physiologic role of this cytokine in host defense using another weakly virulent strain of C. neoformans, which induced the expression of IL-18 mRNA in the infected lungs. Administration of this Ab exacerbated the infection, as shown by the increased lung burden. Our results indicate that IL-18 plays an important role in host defense against infection with C. neoformans.