Crystal structures of multidrug binding protein TtgR in complex with antibiotics and plant antimicrobials.
Antibiotic resistance is a widely spread phenomenon. One major mechanism that underlies antibiotic resistance in bacteria is the active extrusion of toxic compounds through the membrane-bound efflux pumps that are often regulated at the transcriptional level. TtgR represses the transcription of TtgABC, a key efflux pump in Pseudomonas putida, which is highly resistant to antibiotics, solvents and toxic plant secondary products. Previously we showed that TtgR is the only reported repressor that binds to different classes of natural antimicrobial compounds, which are also extruded by the efflux pump. We report here five high-resolution crystal structures of TtgR from the solvent-tolerant strain DOT-T1E, including TtgR in complex with common antibiotics and plant secondary metabolites. We provide structural basis for the unique ligand binding properties of TtgR. We identify two distinct and overlapping ligand binding sites; the first one is broader and consists of mainly hydrophobic residues, whereas the second one is deeper and contains more polar residues including Arg176, a unique residue present in the DOT-T1E strain but not in other Pseudomonas strains. Phloretin, a plant antimicrobial, can bind to both binding sites with distinct binding affinities and stoichiometries. Results on ligand binding properties of native and mutant TtgR proteins using isothermal titration calorimetry confirm the binding affinities and stoichiometries, and suggest a potential positive cooperativity between the two binding sites. The importance of Arg176 in phloretin binding was further confirmed by the reduced ability of phloretin in releasing the mutant TtgR from bound DNA compared to the native protein. The results presented here highlight the importance and versatility of regulatory systems in bacterial antibiotic resistance and open up new avenues for novel antimicrobial development.


Predictive value of bronchoalveolar lavage cell analysis in sarcoidosis.
Patients with histopathologically proved sarcoidosis were studied serially by means of bronchoalveolar lavage, initially at the time of diagnosis and then six and 12 months later. Two years later they were evaluated by chest radiography and lung function tests and classified in terms of recovery or progression over the previous two years. The recovery of lymphocytes and granulocytes in lavage fluid was of limited prognostic value for persistent lung disease. In contrast, patients with increased numbers of mast cells recovered by lavage were more likely to deteriorate. Significantly increased mast cell counts (greater than or equal to 0.5% of total cells recovered) were seen in at least one lavage investigation in 15 of the 16 patients with more active and progressive disease, but in only eight of 23 patients with inactive disease (p less than 0.001). A persistent increase of mast cells on serial measurement occurred in nine of the 16 patients with active disease and in four of the 23 patients in whom the disease was inactive (p less than 0.02). The finding in the two subsequent lavages of lymphocytosis (lymphocytes greater than 30% of recovered cells) or neutrophilia (neutrophils greater than 15%) combined with mastocytosis (mast cells greater than or equal to 0.5%) occurred in nine of the 16 patients with active disease but in no patients with inactive disease.


Pharmacokinetics of Teicoplanin in a Patient with Coronavirus Disease 2019 Receiving Veno-venous Extracorporeal Membrane Oxygenation.
Patients with severe coronavirus disease 2019 (COVID-19) receiving ventilation or pulmonary support via veno-venous extracorporeal membrane oxygenation (VV-ECMO) can be infected with drug-resistant bacteria. When introducing VV-ECMO, the changes in serum antibiotic concentration should be considered due to an increased volume of distribution (Vd). However, no pharmacokinetic study has assessed teicoplanin (TEIC) treatment in patients with COVID-19 receiving VV-ECMO. A 71-year-old man diagnosed with COVID-19 visited a primary hospital. His oxygenation conditions worsened despite treatment with favipiravir and methylprednisolone as well as oxygen therapy. After his transfer to our center, tracheal intubation and steroid pulse therapy were initiated. Seven days after admission, VV-ECMO was performed. TEIC was administered for secondary bacterial infection. The serum TEIC concentration remained within the therapeutic range, indicating that VV-ECMO did not significantly affect TEIC pharmacokinetics. VV-ECMO was discontinued 17 days after admission. However, he developed multi-organ disorder and died 42 days after admission. As TEIC prevents viral invasion, it may be used with ECMO in patients with COVID-19 requiring ventilation; however, the altered pharmacokinetics of TEIC, such as increased Vd, should be considered. Therefore, TEIC pharmacokinetics in VV-ECMO should be assessed in future studies with an appropriate number of patients.


Polymer Brushes on Graphitic Carbon Nitride for Patterning and as a SERS Active Sensing Layer via Incorporated Nanoparticles.
Graphitic carbon nitride (gCN) has a broad range of promising applications, from energy harvesting and storage to sensing. However, most of the applications are still restricted due to gCN poor dispersibility and limited functional groups. Herein, a direct photografting of gCN using various polymer brushes with tailorable functionalities via UV photopolymerization at ambient conditions is demonstrated. The systematic study of polymer brush-functionalized gCN reveals that the polymerization did not alter the inherent structure of gCN. Compared to the pristine gCN, the gCN-polymer composites show good dispersibility in various solvents such as water, ethanol, and tetrahydrofuran (THF). Patterned polymer brushes on gCN can be realized by employing photomask and microcontact printing technology. The polymer brushes with incorporated silver nanoparticles (AgNPs) on gCN can act as a multifunctional recyclable active sensing layer for surface-enhanced Raman spectroscopy (SERS) detection and photocatalysis. This multifunctionality is shown in consecutive cycles of SERS and photocatalytic degradation processes that can be applied to in situ monitor pollutants, such as dyes or pharmaceutical waste, with high chemical sensitivity as well as to water remediation. This dual functionality provides a significant advantage to our AgNPs/polymer-gCN with regard to state-of-the-art systems reported so far that only allow SERS pollutant detection but not their decomposition. These results may provide a new methodology for the covalent functionalization of gCN and may enable new applications in the field of catalysis, biosensors, and, most interestingly, environmental remediation.


Cerebral venous sinus thrombosis: A diagnostic challenge in a rare presentation.
Cerebral venous sinus thrombosis (CVST) is an uncommon, life-threatening condition with a variable clinical presentation that makes it a challenge of diagnosis. A 39-year-old male patient presented to the hospital with complete loss of conscious and admitted to Medical Intensive Care Unit for investigation without any obvious history that was difficult for diagnosis. In this case, the patient presented with coma that is a rare presentation of CVST with no obvious clinical history and he was male patient that means he is free of all gender-specific risk factors of CVST. The brain computed tomography (CT) scan showed hypodense lesion in the left upper parietal region with no hemorrhage. The lesion was low-signal intensity (SI) on T1WIs and high SI on T2WIs and restricted on diffusion-weighted images like arterial infarctions, but magnetic resonance angiography (MRA) was normal that excluded arterial infarction. Gadolinium-enhanced MR venography (MRV) showed the filling defect of CVST. CVST can be present by a mysterious clinical presentation that makes it as a challenge of diagnosis even by medical imaging by CT and MR imaging (MRI). A combination of MRI and MRV is the best, noninvasive, and nonionizing imaging modality for the diagnosis of CVST.


Rapid diagnosis of Indian visceral leishmaniasis using achatininH, a 9-O-acetylated sialic acid binding lectin.
We have developed a rapid, accurate, and inexpensive hemagglutination assay (HA) for diagnosis of Indian visceral leishmaniasis (VL) using achatininH, a 9-O-acetylated sialic acid binding lectin. The assay is based on the selective binding of achatininH to 9-O-acetylated derivatives of sialic acid (9-O-AcSA), which appears exclusively on the erythrocytes of patients with VL. Blood samples from 35 patients with demonstrable splenic or bone marrow amastigotes were assayed by HA and compared with an ELISA. All cases were correctly diagnosed by HA, and were in concordance with the ELISA. The increase in 9-O-AcSA was corroborated by direct fluorimetric estimation of sialic acid and its 9-O-acetylated derivative. Therefore, 9-O-acetylated derivatives of sialic acid, absent in normal erythrocytes, serve as an important biomarker, and indirectly reflect the presence of the parasite. Accordingly, this assay can be used for detection of active kala-azar cases and merits clinical consideration.


Differential effects of ketamine on gating of auditory evoked potentials and prepulse inhibition in rats.
Schizophrenic patients suffer from deficits in information processing. Patients show both a decrease in P50 gating [assessed in the conditioning-testing (C-T) paradigm] and prepulse inhibition (PPI), two paradigms that assess gating. These two paradigms might have a related underlying neural substrate. Gating, as measured in both the C-T paradigm (the gating of a component of the auditory evoked potential (AEP)], and PPI can easily be measured in animals as well as in humans. This offers the opportunity to model these information processing paradigms in animals in order to investigate the effects of neurotransmitter manipulations in the brain. In order to validate the animal model for disturbances in AEP gating, d-amphetamine (0.5 and 1 mg/kg, i.p.) was administered. Gating of an AEP component was changed due to injection of d-amphetamine (1 mg/kg) in the same way as seen in schizophrenic patients: both the amplitude to the conditioning click and the gating were significantly reduced. Next, the effect of the N-methyl-D-aspartate (NMDA) antagonist ketamine (2.5 and 10 mg/kg, i.p.) was investigated to assess its effects in the two gating paradigms. It was found that ketamine (10 mg/kg) did not affect gating as measured with components of the AEP. However, ketamine (10 mg/kg) disrupted PPI of the startle response to the extent that prepulse facilitation occurred. Firstly, it is concluded that AEP gating was disrupted by d-amphetamine and not by ketamine. Secondly, PPI and the C-T paradigm reflect distinct inhibitory sensory processes, since both paradigms are differentially influenced by ketamine.


Neuropsychology and cognitive health in healthy older adults: a brief overview for psychiatric nurses.
This article contains a brief synopsis on nonpathological aspects of the neuropsychology of aging and cognitive health. In nonpathological aging, normal subtle decline occurs in a number of cognitive domains such as executive functioning, speed of processing, memory, language, and psychomotor ability; however, some domains of cognitive functioning appear to increase with age, such as vocabulary and crystallized intelligence. In the neuropsychology and the cognitive aging literatures, several hypotheses for such age-related declines are proposed, including the diminished speed-of-processing hypothesis, the common cause hypothesis, and the frontal aging hypothesis. As these age-related changes diminish cognitive reserve, the decline in the related cognitive domains emerges. Ways to protect and improve cognitive health are suggested to encourage positive neuroplasticity and discourage negative neuroplasticity. Implications for nursing practice are provided.


Flexible electrical recording from cells using nanowire transistor arrays.
Semiconductor nanowires (NWs) have unique electronic properties and sizes comparable with biological structures involved in cellular communication, thus making them promising nanostructures for establishing active interfaces with biological systems. We report a flexible approach to interface NW field-effect transistors (NWFETs) with cells and demonstrate this for silicon NWFET arrays coupled to embryonic chicken cardiomyocytes. Cardiomyocyte cells were cultured on thin, optically transparent polydimethylsiloxane (PDMS) sheets and then brought into contact with Si-NWFET arrays fabricated on standard substrates. NWFET conductance signals recorded from cardiomyocytes exhibited excellent signal-to-noise ratios with values routinely >5 and signal amplitudes that were tuned by varying device sensitivity through changes in water gate-voltage potential, V(g). Signals recorded from cardiomyocytes for V(g) from -0.5 to +0.1 V exhibited amplitude variations from 31 to 7 nS whereas the calibrated voltage remained constant, indicating a robust NWFET/cell interface. In addition, signals recorded as a function of increasing/decreasing displacement of the PDMS/cell support to the device chip showed a reversible >2x increase in signal amplitude (calibrated voltage) from 31 nS (1.0 mV) to 72 nS (2.3 mV). Studies with the displacement close to but below the point of cell disruption yielded calibrated signal amplitudes as large as 10.5 +/- 0.2 mV. Last, multiplexed recording of signals from NWFET arrays interfaced to cardiomyocyte monolayers enabled temporal shifts and signal propagation to be determined with good spatial and temporal resolution. Our modular approach simplifies the process of interfacing cardiomyocytes and other cells to high-performance Si-NWFETs, thus increasing the experimental versatility of NWFET arrays and enabling device registration at the subcellular level.


Recruitment of the extracellular signal-regulated kinase/ribosomal S6 kinase signaling pathway to the NFATc4 transcription activation complex.
Integration of protein kinases into transcription activation complexes influences the magnitude of gene expression. The nuclear factor of activated T cells (NFAT) group of proteins are critical transcription factors that direct gene expression in immune and nonimmune cells. A balance of phosphotransferase activity is necessary for optimal NFAT activation. Activation of NFAT requires dephosphorylation by the calcium-mediated calcineurin phosphatase to promote NFAT nuclear accumulation, and the Ras-activated extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinase, which targets NFAT partners, to potentiate transcription. Whether protein kinases operate on NFAT and contribute positively to transcription activation is not clear. Here, we coupled DNA affinity isolation with in-gel kinase assays to avidly pull down the activated NFAT and identify its associated protein kinases. We demonstrate that p90 ribosomal S6 kinase (RSK) is recruited to the NFAT-DNA transcription complex upon activation. The formation of RSK-NFATc4-DNA transcription complex is also apparent upon adipogenesis. Bound RSK phosphorylates Ser(676) and potentiates NFATc4 DNA binding by escalating NFAT-DNA association. Ser(676) is also targeted by the ERK MAP kinase, which interacts with NFAT at a distinct region than RSK. Thus, integration of the ERK/RSK signaling pathway provides a mechanism to modulate NFATc4 transcription activity.


Globular domains of agrin are functional units that collaborate to induce acetylcholine receptor clustering.
Agrin, an extracellular matrix protein involved in neuromuscular junction formation, directs clustering of postsynaptic molecules, including acetylcholine receptors (AChRs). This activity resides entirely in the C-terminal portion of the protein, which consists of three laminin-like globular domains (G-domains: G1, G2 and G3) and four EGF-like repeats. Additionally, alternate mRNA splicing yields G-domain variants G2(0,4) with 0- or 4-amino-acid inserts, and G3(0, 8,11,19) with 0-, 8-, 11- or 19-amino-acid inserts. In order to better understand the contributions of individual domains and alternate splicing to agrin activity, single G-domains and covalently linked pairs of G-domains were expressed as soluble proteins and their AChR clustering activity measured on cultured C2 myotubes. These analyses reveal the following: (1) While only G3(8) exhibits detectable activity by itself, all G-domains studied (G1, G2(0), G2(4), G3(0) and G3(8)) enhance G3(8) activity when physically linked to G3(8). This effect is most pronounced when G2(4) is linked to G3(8) and is independent of the order of the G-domains. (2) The deletion of EGF-like repeats enhances activity. (3) Increasing the physical separation between linked G1 and G3(8) domains produces a significant increase in activity; similar alterations to linked G2 and G3(8) domains are without effect. (4) Clusters induced by two concatenated G3(8) domains are significantly smaller than all other agrin forms studied. These data suggest that agrin G-domains are the functional units which interact independently of their specific organization to yield AChR clustering. G-domain synergism resulting in biological output could be due to physical interactions between G-domains or, alternatively, independent interactions of G-domains with cell surface receptors which require spatially localized coactivation for optimal signal transduction.


Effects of forestry-driven changes to groundcover and soil moisture on amphibian desiccation, dispersal, and survival.
Over 80% of amphibian species that are declining are forest dependent. Forestry practices are a major cause of forest alterations globally, and it is well documented that clearcutting can contribute to amphibian declines. However, there might be adverse effects of forestry practices other than clearcutting. For example, planting overstory trees in rows (plantations) can change groundcover microhabitats and soil moisture levels, but the effects of this common practice on amphibian populations are not well studied. We compared the impacts of common intensive pine plantation operations to naturally regenerated pine forests on the desiccation, movement rates, behavior, and survival of >900 juvenile southern toads (Anaxyrus terrestris). Pine plantations had significantly more accumulation of conifer needles and less exposed soil, herbaceous groundcover, broadleaf litter, and soil moisture than natural pine forests despite the greater canopy cover at plantations. Litter cover explained 85% of groundcover microhabitat variance among forest types and predicted minimum soil moisture levels. When toads were held in small outdoor enclosures that constrained microhabitat selection, 24-h desiccation rates and 72-h mortality were significantly greater in pine plantation than in naturally regenerated pine forest because of lower soil moisture, especially during low rainfall periods. In large outdoor pens where juvenile amphibians could select microhabitats, movement was strongly directed down slope and increased with precipitation. However, initial speeds were positively associated with pine density, likely because toads were trying to evacuate from the drier high-pine-density areas. High-intensity silviculture practices that eliminate herbaceous or vegetative groundcover, such as roller chopping and scalping, increase amphibian desiccation because planted conifers dry the upper soil layer. Our study highlights the importance of prioritizing lower intensity silviculture practices or lower pine densities to retain groundcover microhabitat that serves as amphibian refugia from dry conditions that are predicted to increase in frequency with climate change.


Information of imminent death or not: does it make a difference?
This study examines whether end-of-life care for patients with cancer who were informed about imminent death differs from care for those patients with cancer who were not informed. This study included all cancer deaths between 2006 and 2008 for which the patient did not lose his or her decision-making capacities until hours or days before death (N=13,818). These patients were taken from a national quality register for end-of-life care. The majority of the patients-91% (n=12,609) -had been given information about imminent death; 9% (n=1,209) had not been informed. Because of the difference in sample size, a matching procedure was performed to minimize bias. This resulted in a comparison of 1,191 informed and 1,191 uniformed patients. Nonparametric methods were used for statistical analyses. Informed patients significantly more often had parenteral drugs prescribed as needed (ie, PRN), had his or her family informed, died in his or her preferred place, and had family who were offered bereavement support. There was no difference in symptom control (ie, pain, anxiety, confusion, nausea, and respiratory tract secretions) between the groups. Providing information of imminent death to a patient with cancer at the end of life does not seem to increase pain or anxiety, but it does seem to be associated with improved care and to increase the likelihood of fulfilling the principles of a good death.


Evaluating commercially available rodenticide baits for invasive Gambian giant pouched rats (Cricetomys gambianus).
Gambian giant pouched rats (Cricetomys gambianus) are native to Africa, but they are popular in the pet industry in the United States. They were reservoir hosts during a monkeypox outbreak in the Midwestern United States in 2003. A free-ranging population became established on Grassy Key in the Florida Keys, apparently because of a release by a pet breeder. These rodents could cause significant damage to agricultural crops should they reach the mainland. Research under controlled conditions was needed to identify effective rodenticides for Grassy Key or other cases where an invasion of Gambian rats might occur. We tested 2 formulations of diphacinone baits and 1 formulation each of brodifacoum, zinc phosphide, bromethalin, and chlorophacinone baits with captive Gambian rats in multiple-choice food trials. Both the brodifacoum and zinc phosphide rodenticide baits were highly effective (100% mortality). Also, brodifacoum and zinc phosphide treatments performed similar to the Environmental Protection Agency's standard for toxicants of (i.e., 90% mortality in laboratory trials). The chlorophacinone, diphacinone, and bromethalin baits did not appear to be very effective at killing Gambian rats (≤50% mortality) in our study. Effective tools to combat Gambian giant pouched rats have been identified in a laboratory trial. Further field testing of commercially available brodifacoum and zinc phosphide baits may prove useful for controlling the potentially invading Gambian rats.


Diagnosis and management of venereal campylobacteriosis in beef cattle.
Bovine venereal campylobacteriosis is caused by Campylobacter fetus subsp. venerealis and its glycerine-tolerant variant Campylobacter fetus subsp. venerealis biovars intermedius. The disease can be economically important when present in cattle herds, causing poor reproductive performance, embryo mortality and abortion. Sensitive and specific diagnostic tests are required in the diagnosis of infection and to inform and monitor disease control. Current tests include bacterial culture and fluorescent antibody testing of preputial sheath washings and an enzyme-linked immunosorbent assay and an agglutination test on vaginal mucus, although the predictive values of these tests can be inadequate in field investigations. Artificial insemination is often considered as a simple control method for bovine venereal campylobacteriosis, but is impractical for many beef suckler herds where breeding takes place at pasture. Commercial vaccines are unavailable in the UK, while the efficacy of autogenous vaccines using a bacterial isolate from infected animals on a specific farm is at best unproven. Hence, for some infected herds, the development of an alternative control strategy based on segregation of potentially infected and uninfected animals in combination with culling or treatment would be desirable. This approach requires meticulous records and herd health management. In this paper we highlight difficulties in diagnosing bovine venereal campylobacteriosis and demonstrate the benefits of good record keeping when investigating poor reproductive performance in a beef suckler herd and establishing a herd-specific approach to bio-containment of the infectious cause. Bovine venereal campylobacteriosis is an economically important disease that should be considered in investigations of suckler herd subfertility problems. Control of the disease based on segregation of potentially infected and uninfected animals in combination with extensive culling can be achieved without the use of artificial insemination or vaccination, but requires meticulous records and strict adherence to herd biosecurity practices.


Chronic use of tramadol after acute pain episode: cohort study.
To determine the risk of prolonged opioid use in patients receiving tramadol compared with other short acting opioids. Observational study of administrative claims data. United States commercial and Medicare Advantage insurance claims (OptumLabs Data Warehouse) January 1, 2009 through June 30, 2018. Opioid-naive patients undergoing elective surgery. Risk of persistent opioid use after discharge for patients treated with tramadol alone compared with other short acting opioids, using three commonly used definitions of prolonged opioid use from the literature: additional opioid use (defined as at least one opioid fill 90-180 days after surgery); persistent opioid use (any span of opioid use starting in the 180 days after surgery and lasting ≥90 days); and CONSORT definition (an opioid use episode starting in the 180 days after surgery that spans ≥90 days and includes either ≥10 opioid fills or ≥120 days' supply of opioids). Of 444 764 patients who met the inclusion criteria, 357 884 filled a discharge prescription for one or more opioids associated with one of 20 included operations. The most commonly prescribed post-surgery opioid was hydrocodone (53.0% of those filling a single opioid), followed by short acting oxycodone (37.5%) and tramadol (4.0%). The unadjusted risk of prolonged opioid use after surgery was 7.1% (n=31 431) with additional opioid use, 1.0% (n=4457) with persistent opioid use, and 0.5% (n=2027) meeting the CONSORT definition. Receipt of tramadol alone was associated with a 6% increase in the risk of additional opioid use relative to people receiving other short acting opioids (incidence rate ratio 95% confidence interval 1.00 to 1.13; risk difference 0.5 percentage points; P=0.049), 47% increase in the adjusted risk of persistent opioid use (1.25 to 1.69; 0.5 percentage points; P<0.001), and 41% increase in the adjusted risk of a CONSORT chronic opioid use episode (1.08 to 1.75; 0.2 percentage points; P=0.013). People receiving tramadol alone after surgery had similar to somewhat higher risks of prolonged opioid use compared with those receiving other short acting opioids. Federal governing bodies should consider reclassifying tramadol, and providers should use as much caution when prescribing tramadol in the setting of acute pain as for other short acting opioids.


The Effects of Hand Massage on Stress and Agitation Among People with Dementia in a Hospital Setting: A Pilot Study.
Agitation in people with dementia is a growing concern as it causes distress for both patients and their nurses and may contribute to relational disorders. Previous studies involving patients with dementia living in long-term care facilities have reported decreased agitation following massage. The objective of this pilot study was to investigate the effect of hand massage on agitation and biological markers of stress in patients with dementia hospitalized in an acute geriatric psychiatry service. In this randomized controlled trial we included 40 agitated patients with dementia with an intervention group and a control group. The study is designed to test the effect of seven hand massages over three continuous weeks on agitation and levels of salivary cortisol (sC) and alpha-amylase (sAA). Compared to the control group, the intervention group exhibited larger increases in sC and sAA at week 1 from before to after the massage, but larger decreases at week 2 and 3, with a significant group effect for sAA at week 2. Agitation scores were not significantly different between the groups but tended to decrease more in the intervention group than the control group. This study provides first encouraging results suggesting that hand massage might have beneficial effects on stress and agitation in hospitalized patients with dementia. It also highlights the challenges associated with conducting such studies with this complex patient population. Further studies are needed to confirm these findings and the benefits of hand massage as part of routine care for patients with dementia.


Diode-pumped 1.7-W erbium 3-mum fiber laser.
We report what is to our knowledge the first 3-mu;m fiber laser of the 1-W class. 1.7 W of output power and 17.3% slope efficiency (with respect to the launched pump power) at a wavelength of 2.71mum are demonstrated from a double-clad erbium-doped ZBLAN fiber diode pumped at 790 nm. Energy transfer from the Er(3+) lower laser level to a Pr(3+) codopant decreases ground-state bleaching and excited-state absorption, thus avoiding output-power saturation. This result represents more than an order-of-magnitude improvement over previous work of which we are aware. Advantages over current crystal-laser designs include nearly transverse-fundamental-mode operation, reduced thermal effects, and ease of use, e.g., in medical endoscopy.


Adaptive mutations in PB2 gene contribute to the high virulence of a natural reassortant H5N2 avian influenza virus in mice.
The highly pathogenic A/chicken/Hebei/1102/2010 (HB10) H5N2 virus is a natural reassortant derived from circulating H5N1 and endemic H9N2 avian influenza viruses (AIV). To evaluate the potential of its interspecies transmission, we previously serially passaged the non-virulent HB10 virus in the mouse lung and obtained a high virulence variant (HB10-MA). Genomic sequencing revealed five mutations (HA-S227N, PB2-Q591K, PB2-D701N, PA-I554V and NP-R351K) that distinguished HB10-MA virus from its parental HB10 virus. In this study, we further investigated the molecular basis for the enhanced virulence of HB10-MA in mice. By generating a series of reassortants between the two viruses and evaluating their virulence in mice, we found that both PB2 and PA genes contribute to the high virulence of HB10-MA in mice, whereas PB2 gene carrying the 591K and/or 701N had a dominant function. In addition, the two amino acids showed a cumulative effect on the virulence, virus replication, and polymerase activity of HB10 or HB10-MA. Therefore, our results collectively emphasized the crucial role of PB2 gene, particularly the paired mutations of Q591K and D701N in the host adaptation of the novel reassortant H5N2 AIV in mammals, which may provide helpful insights into the pathogenic potential of emerging AIV in human beings.


The Combined Effect of Cigarette Smoking and Fitness on Injury Risk in Men and Women.
Prior studies have identified cigarette smoking and low fitness as independent risk factors for injury; however, no studies have evaluated the combined effect of cigarette smoking and fitness on injury risk. To evaluate the combined effect of cigarette smoking and fitness on injury risk in men and women. This is a secondary analysis of data collected from US Army recruits (n = 2000) during basic combat training within the United States in 2007. Physical training and fitness, cigarette smoking, and prior injury data were obtained from questionnaires, whereas demographic and injury data were obtained from medical and basic combat training unit records. Chi-squared tests were used to assess differences in injury risk by fitness level and cigarette smoking. Relative risk values were calculated with 95% confidence intervals. The primary findings showed that smokers experienced 20%-30% higher risk of injury than nonsmokers. In addition, higher aerobic and muscular fitness was generally not protective against injury between least fit and more fit smokers. However, higher fitness was protective against injury between least and more fit nonsmokers, with least fit nonsmokers being 30%-50% more likely to experience an injury than fit nonsmokers. This study revealed that higher aerobic and muscular fitness was not protective against injury among smokers; however, it was protective against injury among nonsmokers. Further implementation of smoking cessation programs may be beneficial for military and civilian personnel who are required to be physically fit in order to carry out their job responsibilities. Male and female smokers experienced significantly higher risk of injury than nonsmokers. Although higher fitness is protective against injury in nonsmokers, the protective effect of fitness is lost among smokers. In an attempt to reduce injury risk among military and emergency personnel, smoking cessation programs should be further implemented among both more fit and less fit smokers.


Lack of hypoxia-inducible factor-1 alpha impairs midbrain neural precursor cells involving vascular endothelial growth factor signaling.
Oxygen tension is critical for proliferation of human and murine midbrain-derived neural precursor cells (mNPCs). Here, we conditionally inactivated the hypoxia-responsive transcription factor hypoxia-inducible factor-1alpha (HIF-1alpha) in murine NPCs to determine its role in proliferation, survival, and dopaminergic differentiation in vitro as well as survival of murine dopaminergic neurons in vivo. HIF-1alpha conditional knock-out (HIF-1alpha CKO) mNPCs showed midbrain-specific impairment of survival and proliferation. Dopaminergic differentiation of HIF-1alpha CKO mNPCs in vitro was markedly reduced. Expression of vascular endothelial growth factor (VEGF) mRNA was reduced in HIF-1alpha CKO mNPCs, whereas erythropoietin signaling was not affected. Treatment of HIF-1alpha CKO mNPCs with 50 ng/ml VEGF partially recovered proliferation and dopaminergic differentiation in vitro. In substantia nigra (SN) of adult HIF-1alpha CKO mice, protein levels of dopaminergic marker molecules such as tyrosine hydroxylase (TH) and aldehyde dehydrogenase were reduced by 41 and 61%, respectively. The cell survival marker Bcl-2 was reduced by 58% while caspase-3 was activated. Nonbiased stereological cell counts of TH-positive neurons in SN of young adult HIF-1alpha CKO mice revealed a reduction of 31% compared with cre/wt mice (in which the wild-type Hif1a allele is expressed in parallel with the Cre recombinase allele). However, we found no impairment of striatal dopamine concentrations or locomotor behavior. In conclusion, HIF-1alpha seems to be a transcription factor relevant to the development and survival of substantia nigra dopaminergic neurons involving VEGF signaling.


Dissection of the anterior abdominal wall and the deep inguinal region from a laparoscopic perspective.
The usual dissection by medical students of the anterior abdominal wall and the inguinal region proceeds from superficial to deep; special emphasis is placed on the sheath of the rectus abdominis muscle and lateral muscular layers. We suggest an alternate approach to dissection of this region that has the following advantages: (1) sparing of delicate deep structures not often fully appreciated by students; (2) provision of an opportunity to visualize the region from a laparoscopic surgeon's vantage point; (3) considerably reduced time spent dissecting and identifying structures and relationships, especially peritoneal reflections important in laparoscopic procedures. Our dissection begins with bilateral subcostal incisions through the entire thickness of the anterior abdominal wall and peritoneum, which extend laterally and inferiorly to the level of the anterior superior iliac spines, thereby forming a large, inverted, U-shaped flap. This flap is reflected inferiorly, allowing abdominal viscera to be dissected, and ultimately removed en bloc. The flap is then drawn cranially and stretched somewhat to approximate its position when the abdomen is inflated with CO2 during laparoscopic procedures. Major landmarks, including the deep inguinal ring, are noted and the flap is again reflected inferiorly for dissection beginning with the peritoneum and transversalis fascia. This method of dissecting the anterior abdominal wall and inguinal region results in more facile and timely identification of both superficial and deep structures of the anterior abdominal wall and inguinal region, and provides a clinically relevant demonstration of anatomy from a laparoscopic perspective.


Experimental realization of a mode-locked parabolic Raman fiber oscillator.
We report here the first demonstration of a mode-locked fiber laser delivering parabolic pulses (similaritons) at 1534 nm. The use of a Raman-based gain medium potentially allows its implementation at any wavelength. The 22nJ output similariton pulses have a true parabolic shape both in the time and spectral domains and a linear chirp. Linear recompression close to Fourier limit is demonstrated allowing us to obtain 6 ps compressed pulses with a compression factor of 75.


Racial differences in prognostic value of adult height for biochemical progression following radical prostatectomy.
Adult height, as a surrogate of childhood and adolescent hormone activity and diet, has been associated with the risk for development and death from prostate cancer in predominantly White populations. However, hormonal activity and diets vary between races. We examined whether height was significantly associated with biochemical progression following radical prostatectomy and whether there was an interaction between height and race. Multivariate Cox proportional hazards analysis was used to determine if height significantly predicted biochemical progression among 1,503 men (450 Black and 1,053 White) treated with radical prostatectomy between 1988 and 2003. We examined for possible interactions between height and race. Taller men (>175.3 cm) were significantly younger (P = 0.001), treated in more recent years (P = 0.02), had more clinical stage T(1) disease (P = 0.001), and were less likely to have extraprostatic extension (P = 0.02) than shorter men (< or =175.3 cm). Height was not significantly related to race, preoperative serum prostate-specific antigen concentrations, biopsy or pathologic Gleason sum, positive surgical margins, seminal vesicle invasion, or lymph node metastasis. Height was significantly associated with progression among Black men [relative risk (RR), 1.67; 95% confidence interval (95% CI), 1.00-2.79] but not among White men (RR, 1.03; 95% CI, 0.77-1.38). The interaction between race and height for predicting biochemical progression was statistically significant (P(interaction) = 0.05). There was an interaction between height and race in that height predicted progression for Black men but not for White men. The explanation for these findings is unclear, although lower insulin-like growth factor-binding protein-3 concentrations among Black men may be involved.


Practices and Perceptions of Animal Contact and Associated Health Outcomes in Pregnant Women and New Mothers.
Companion animals play an important role in our society. However, pregnant women and new mothers might have specific concerns about animal-associated health outcomes because of their altered immune function and posture as well as their newborn babies. The study was conducted to collect baseline data for developing an evidence-based intervention for pregnant women and new mothers to help them adopt certain behaviors to prevent adverse animal-associated health outcomes. A survey, using the Health Belief Model as the theoretical framework, was developed and administered to 326 women attending the Women, Infants, and Children programs in Illinois and Indiana in 2015. Prevalence of dog and cat ownership was estimated to be 39% (95% CI: 33-45%) and 26% (95% CI: 21-31%), respectively. Regardless of pet ownership, 74% of the respondents reported having some type of animal contact in the past month. Pregnancy or the birth of a child altered some animal contact practices among the study participants; particularly a discontinuation or decrease in cleaning cat litter boxes. Reports of diseases contracted from animals were low (4%) in this study. By contrast, animal-associated injuries were prevalent (42%), and the majority were caused by animals the respondents owned (56%). Overall, respondents indicated that they appreciated the benefits of a program addressing animal-associated health outcomes and did not indicate strong resistance to adopt certain behaviors. The majority recognized human health-care providers as a source of information about animal contact and associated health outcomes but less frequently identified veterinarians as a source for such information. In addition, although most of the respondents felt that health-care providers and veterinarians should initiate discussions about preventing animal-associated illness and injuries, only 41% among those who had visited doctors or prenatal care services reported that their health-care providers discussed these issues with them. The results indicate the importance of an intervention concerning animal contact and associated health outcomes for the target population and provide insights to the potential implications of program implementation.


Gluten-free diet does not appear to induce endoscopic remission of eosinophilic esophagitis in children with coexistent celiac disease.
Celiac disease and eosinophilic esophagitis are usually considered to be separate gastrointestinal diseases; however, it appears that they may coexist more often than would be expected. It is unknown whether eosinophilic esophagitis in patients with celiac disease responds to a gluten-free diet. To examine the clinical, endoscopic and histological features of children with both conditions to evaluate whether eosinophilic esophagitis responds to a gluten-free diet. From January 1, 2009, to June 30, 2011, the medical records of children <18 years of age diagnosed with eosinophilic esophagitis and⁄or celiac disease were reviewed. Patients with clinical, endoscopic and histological diagnoses of both diseases were identified and included. These findings were analyzed, as were laboratory results, treatment and follow-up. During the study period, there were 206 celiac disease patients, 86 eosinophilic esophagitis patients and nine (4.4% of total celiac) patients with both diagnoses. Gluten-free diet was the primary treatment for both conditions in seven of nine (78%) cases. In six of these seven (86%) patients, no endoscopic or histological improvement of eosinophilic esophagitis was observed, while in one patient, histological remission of esophageal eosinophilia occurred while on a gluten-free diet. The prevalence of eosinophilic esophagitis in patients with celiac disease was 4.4%, confirming a higher than expected prevalence of eosinophilic esophagitis compared with the general population. In patients with celiac disease, a gluten-free diet did not appear to induce remission of coexistent endoscopic and histological features of eosinophilic esophagitis.


Enhancement of tick-borne encephalitis virus transmission by tick salivary gland extracts.
To investigate the role of ticks in TBE virus transmission, salivary gland extract (SGE) was derived from partially fed female Ixodes ricinus, Dermacentor reticulatus and Rhipicephalus appendiculatus ticks. Guinea-pigs were infested with uninfected R. appendiculatus nymphs and inoculated with a mixture of TBE virus and SGE or with virus alone. The number of ticks which on average acquired virus from feeding on animals inoculated with TBE virus and SGE from partially fed ticks was 4-fold greater than the number that became infected by feeding on animals inoculated with virus alone or virus plus SGE from unfed I. ricinus. Viraemia was detected in 67% of guinea-pigs inoculated with virus plus SGE compared to 30% of guinea-pigs inoculated with virus alone. Virus titres in the blood were similar for both groups of animals [range 2.0-2.8 log10 plaque-forming units (PFU)/ml of blood]; however, the number of ticks that became infected was significantly higher on animals inoculated with virus plus SGE from partially fed ticks. No significant difference was observed with respect to the tick species used to derive SGE. The results indicate that TBE virus transmission is enhanced by factor(s) associated with the salivary glands of feeding ticks, and that these factor(s) may facilitate efficient transmission of TBE virus between infected and uninfected ticks even when they feed on hosts that have no detectable viraemia.


Efficient elimination of pancreatic cancer stem cells by hedgehog/GLI inhibitor GANT61 in combination with mTOR inhibition.
Pancreatic cancer is one of the most lethal malignancies. The innovative treatments are required and now the cancer stem cells (CSCs) are expected to be an effective target for novel therapies. Therefore we investigated the significance of hedgehog (Hh) signaling in the maintenance of CSC-like properties of pancreatic cancer cells, in order to discover the key molecules controlling their unique properties. Human pancreatic cancer cell lines, Capan-1, PANC-1, MIA PaCa-2 and Capan-1 M9 were used for our experiments in DMEM/F12 medium containing 10 % fetal bovine serum. Sphere formation assay, immunofluorescence staining, flow cytometric analysis and MTT cell viability assay were performed to investigate molecular signals and the efficacy in the treatment of pancreatic cancer cells. Inhibition of the Hh pathway significantly reduced the expression of stem cell marker CD133 and sphere formation, an index of self-renewal capacity, demonstrating the suppression of CSC-like properties. Moreover, the GLI inhibitor GANT61 induced greater reduction in sphere formation and cell viability of pancreatic cancer cells than the smoothened (SMO) inhibitor cyclopamine. This suggests that GLI transcription factors, but not SMO membrane protein, are the key molecules in the Hh pathway. The treatment using GANT61 in combination with the inhibition of mTOR, which is another key molecule in pancreatic CSCs, resulted in the efficient reduction of cell viability and sphere formation of an inhibitor-resistant cell line, showing the strong efficacy and wide range applicability to pancreatic CSC-like cells. Thus, this novel combination treatment could be useful for the control of pancreatic cancer by targeting pancreatic CSCs. This is the first report of the efficient elimination of pancreatic cancer stem-like cells by the double blockage of Hh/GLI and mTOR signaling.


Lymph node - an organ for T-cell activation and pathogen defense.
The immune system is a multicentered organ that is characterized by intimate interactions between its cellular components to efficiently ward off invading pathogens. A key constituent of this organ system is the distinct migratory activity of its cellular elements. The lymph node represents a pivotal meeting point of immune cells where adaptive immunity is induced and regulated. Additionally, besides barrier tissues, the lymph node is a critical organ where invading pathogens need to be eliminated in order to prevent systemic distribution of virulent microbes. Here, we explain how the lymph node is structurally and functionally organized to fulfill these two critical functions - pathogen defense and orchestration of adaptive immunity. We will discuss spatio-temporal aspects of cellular immune responses focusing on CD8 T cells and review how and where these cells are activated in the context of viral infections, as well as how viral antigen expression kinetics and different antigen presentation pathways are involved. Finally, we will describe how such responses are regulated and 'helped', and discuss how this relates to intranodal positioning and cellular migration of the various cellular components that are involved in these processes.


Preparation of protein nano-objects by assembly of polymer-grafted proteins.
We carried out surface-grafting from proteins and their assembling into objects with unique nanostructured materials (nano-objects). To immobilize polymer-initiating sites, amino groups of bovine serum albumin (BSA) were allowed to react with iniferter groups (BSA-i). Then, graft polymerization of N-isopropyl acrylamide (NIPAM) was performed by light-initiated living radical polymerization from immobilized iniferter moieties of BSA-i. The polymer-grafted BSA (BSA-g-PNIPAM) was assembled into nano-objects through the precipitation of PNIPAM graft chains and their sizes and morphologies were tuned by the chain length, the density and the chemical structure of graft polymers in addition to the environmental conditions such as temperature and pH. It was possible to retain the structures of nano-objects by thermal denaturation via heat treatment. Fluorescent substances were encapsulated in particulate nano-objects (nanoparticles) assembled from PNIPAM-g-BSA and their release could be regulated by tuning pH and temperature. Next, further graft polymerization from PNIPAM-grafted BSA was carried out by living radical polymerization of a cationic monomer, N,N-dimethylamino propyl acrylamide methyl chloride quaternary (DMAPAAQ). The grafted polymer was composed of a block copolymer of PNIPAM and a cationic polymer (PDMAPAAQ) and the gel-like nano-object was generated by increasing temperature. In contrast to PNIPAM-g-BSA, it became insoluble even when the temperature decreased, probably due to the electrostatic association between anionic regions of BSA and cationic regions of graft polymers. Coating of BSA-g-P(NIPAM-b-DMAPAAQ) enabled to form a uniform thin layer over a human hair. A free-standing membrane could be obtained by peeling from a water repellent substrate to create a porous membrane.


Antimutagenic and antiherpetic activities of different preparations from Uncaria tomentosa (cat's claw).
Uncaria tomentosa have been used to treat viral diseases such as herpes due to multiple pharmacological effects, but its therapeutic efficacy against this virus have not been reported yet. Thus, in vitro antiherpetic activity of hydroethanolic extract from barks, purified fractions of quinovic acid glycosides and oxindole alkaloids was evaluated by plaque reduction assay, including mechanistic studies (virucidal, attachment and penetration action). Once exposure to physical agents might lead to reactivation of the herpetic infection, antimutagenic effect (pre-, simultaneous and post-treatment protocols) was also evaluated by Comet assay. The antiherpetic activity from the samples under investigation seemed to be associated with the presence of polyphenols or their synergistic effect with oxindole alkaloids or quinovic acid glycosides, once both purified fractions did not present activity when evaluated alone. Inhibition of viral attachment in the host cells was the main mechanism of antiviral activity. Although both purified fractions displayed the lowest antimutagenic activity in pre and simultaneous treatment, they provided a similar effect to that of cat's claw hydroethanolic extract in post-treatment. Given that purified fractions may result in a reduced antiherpetic activity, the use of cat's claw hydroethanolic extract from barks should be prioritized in order to obtain a synergistic effect.


Antidepressant use and risk of incident cardiovascular morbidity and mortality among postmenopausal women in the Women's Health Initiative study.
Antidepressants are commonly prescribed medications, but their effect on cardiovascular morbidity and mortality remains unclear. Prospective cohort study of 136 293 community-dwelling postmenopausal women in the Women's Health Initiative (WHI). Women taking no antidepressants at study entry and who had at least 1 follow-up visit were included. Cardiovascular morbidity and all-cause mortality for women with new antidepressant use at follow-up (n = 5496) were compared with those characteristics for women taking no antidepressants at follow-up (mean follow-up, 5.9 years). Antidepressant use was not associated with coronary heart disease (CHD). Selective serotonin reuptake inhibitor (SSRI) use was associated with increased stroke risk (hazard ratio [HR],1.45, [95% CI, 1.08-1.97]) and all-cause mortality (HR,1.32 [95% CI, 1.10-1.59]). Annualized rates per 1000 person-years of stroke with no antidepressant use and SSRI use were 2.99 and 4.16, respectively, and death rates were 7.79 and 12.77. Tricyclic antidepressant (TCA) use was associated with increased risk of all-cause mortality (HR,1.67 [95% CI, 1.33-2.09]; annualized rate, 14.14 deaths per 1000 person-years). There were no significant differences between SSRI and TCA use in risk of any outcomes. In analyses by stroke type, SSRI use was associated with incident hemorrhagic stroke (HR, 2.12 [95% CI, 1.10-4.07]) and fatal stroke (HR, 2.10 [95% CI, 1.15-3.81]). In postmenopausal women, there were no significant differences between SSRI and TCA use in risk of CHD, stroke, or mortality. Antidepressants were not associated with risk of CHD. Tricyclic antidepressants and SSRIs may be associated with increased risk of mortality, and SSRIs with increased risk of hemorrhagic and fatal stroke, although absolute event risks are low. These findings must be weighed against quality of life and established risks of cardiovascular disease and mortality associated with untreated depression.


Validation of an instrument to assess evidence-based practice knowledge, attitudes, access, and confidence in the dental environment.
This article reports the validation of an assessment instrument designed to measure the outcomes of training in evidence-based practice (EBP) in the context of dentistry. Four EBP dimensions are measured by this instrument: 1) understanding of EBP concepts, 2) attitudes about EBP, 3) evidence-accessing methods, and 4) confidence in critical appraisal. The instrument-the Knowledge, Attitudes, Access, and Confidence Evaluation (KACE)-has four scales, with a total of thirty-five items: EBP knowledge (ten items), EBP attitudes (ten), accessing evidence (nine), and confidence (six). Four elements of validity were assessed: consistency of items within the KACE scales (extent to which items within a scale measure the same dimension), discrimination (capacity to detect differences between individuals with different training or experience), responsiveness (capacity to detect the effects of education on trainees), and test-retest reliability. Internal consistency of scales was assessed by analyzing responses of second-year dental students, dental residents, and dental faculty members using Cronbach coefficient alpha, a statistical measure of reliability. Discriminative validity was assessed by comparing KACE scores for the three groups. Responsiveness was assessed by comparing pre- and post-training responses for dental students and residents. To measure test-retest reliability, the full KACE was completed twice by a class of freshman dental students seventeen days apart, and the knowledge scale was completed twice by sixteen faculty members fourteen days apart. Item-to-scale consistency ranged from 0.21 to 0.78 for knowledge, 0.57 to 0.83 for attitude, 0.70 to 0.84 for accessing evidence, and 0.87 to 0.94 for confidence. For discrimination, ANOVA and post hoc testing by the Tukey-Kramer method revealed significant score differences among students, residents, and faculty members consistent with education and experience levels. For responsiveness to training, dental students and residents demonstrated statistically significant changes, in desired directions, from pre- to post-test. For the student test-retest, Pearson correlations for KACE scales were as follows: knowledge 0.66, attitudes 0.66, accessing evidence 0.74, and confidence 0.76. For the knowledge scale test-retest by faculty members, the Pearson correlation was 0.79. The construct validity of the KACE is equivalent to that of instruments that assess similar EBP dimensions in medicine. Item consistency for the knowledge scale was more variable than for other KACE scales, a finding also reported for medically oriented EBP instruments. We conclude that the KACE has good discriminative validity, responsiveness to training effects, and test-retest reliability.


The dance of the honeybee: how do honeybees dance to transfer food information effectively?
A honeybee informs her nestmates of the location of a flower she has visited by a unique behavior called a "waggle dance." On a vertical comb, the direction of the waggle run relative to gravity indicates the direction to the food source relative to the sun in the field, and the duration of the waggle run indicates the distance to the food source. To determine the detailed biological features of the waggle dance, we observed worker honeybee behavior in the field. Video analysis showed that the bee does not dance in a single or random place in the hive but waggled several times in one place and then several times in another. It also showed that the information of the waggle dance contains a substantial margin of error. Angle and duration of waggle runs varied from run to run, with the range of +/-15 degrees and +/-15%, respectively, even in a series of waggle dances of a single individual. We also found that most dance followers that listen to the waggle dance left the dancer after one or two sessions of listening.


Risk of selected eye diseases in people admitted to hospital for hypertension or diabetes mellitus: record linkage studies.
Associations among hypertension, diabetes mellitus and some ophthalmic diseases are well established; associations with others are more equivocal. The aim was to quantify associations accurately using large epidemiological datasets. Analysis of the Oxford Record Linkage Study (ORLS), 1963-1998, and English linked hospital episode statistics (LHES), 1999-2010; calculation of rate ratios of eye disease in a hypertension cohort and a diabetes cohort, compared with a reference cohort as control. Risk of cataract following hypertension was marginally elevated (rate ratio ORLS 1.15, 95% CI 1.00 to 1.31; LHES 1.06, 1.01 to 1.10), as was risk of glaucoma (LHES 1.07, 1.00 to 1.14) and age-related macular degeneration (AMD) (LHES 1.14, 1.02 to 1.27). Risk of retinal vein or artery occlusion was elevated three- to fivefold in both populations. Risk of retinal detachment was elevated in LHES at 1.52 (1.43 to 1.73). Risk of cataract in diabetes was high in ORLS and LHES at, respectively, 2.95 (2.75 to 3.16) and 2.30 (2.24 to 2.35), as was risk of glaucoma: 2.47 (2.14 to 2.84) and 2.23 (2.15 to 2.30). Risks were high for AMD (10.3, 8.1 to 13.1, and 3.46, 3.35 to 3.58) and retinal detachment (3.41, 2.71 to 4.25, and 7.96, 7.63 to 8.30), and very high for retinal vein and artery occlusion. With the exception of retinal vascular occlusion, elevations of risk of the ophthalmic diseases studied in hypertension were modest. By contrast, there were significant and substantial increases of risk for each eye disease in people with diabetes.


Developing judgments about peers' obligation to intervene.
In some contexts, punishment is seen as an obligation limited to authority figures. In others, it is also a responsibility of ordinary citizens. In two studies with 4- to 7-year-olds (n = 232) and adults (n = 76), we examined developing judgments about whether certain individuals, either authority figures or peers, are obligated to intervene (Study 1) or to punish (Study 2) after witnessing an antisocial action. In both studies, children and adults judged authority figures as obligated to act, but only younger children judged ordinary individuals as also obligated to do so. Taken together, the present findings suggest that younger children, at least in the United States, start off viewing norm enforcement as a universal responsibility, entrusting even ordinary citizens with a duty to intervene in response to antisocial individuals. Older children and adults, though, see obligations as role-dependent-only authority figures are obligated to intervene.


The sEDA(=) and pEDA(=) descriptors of the double bonded substituent effect.
New descriptors of the double bonded substituent effect, sEDA(=) and pEDA(=), were constructed based on quantum chemical calculations and NBO methodology. They show to what extent the σ and π electrons are donated to or withdrawn from the substituted system by a double bonded substituent. The new descriptors differ from descriptors of the classical substituent effect for which the pz orbital of the ipso carbon atom is engaged in the π-electron system of the two neighboring atoms in the ring. For double bonded substituents, the pz orbital participates in double bond formation with only one external atom. Moreover, the external double bond forces localization of the double bond system of the ring, significantly changing the core molecule. We demonstrated good agreement between our descriptors and the Weinhold and Landis' "natural σ and π-electronegativities": so far only descriptors allowing for evaluation of the substitution effect by a double bonded atom. The equivalency between descriptors constructed for 5- and 6-membered model structures as well as linear dependence/independence of the constructed parameters was discussed. Some interrelations between sEDA(=) and pEDA(=) and the other descriptors of (hetero)cyclic systems such as aromaticity and electron density in the ring and bond critical points were also examined.


Nitrate removal in a riparian wetland of the Appalachian Valley and Ridge physiographic province.
Riparian zones within the Appalachian Valley and Ridge physiographic province are often characterized by localized variability in soil moisture and organic carbon content, as well as variability in the distribution of soils formed from alluvial and colluvial processes. These sources of variability may significantly influence denitrification rates. This investigation studied the attenuation of nitrate (NO3- -N) as wastewater effluent flowed through the shallow ground water of a forested headwater riparian zone within the Appalachian Valley and Ridge physiographic province. Ground water flow and NO3- -N measurements indicated that NO3- -N discharged to the riparian zone preferentially flowed through the A and B horizons of depressional wetlands located in relic meander scars, with NO3- -N decreasing from > 12 to < 0.5 mg L(-1). Denitrification enzyme activity (DEA) attributable to riparian zone location, soil horizon, and NO3- -N amendments was also determined. Mean DEA in saturated soils attained values as high as 210 microg N kg(-1) h(-1), and was significantly higher than in unsaturated soils, regardless of horizon (p < 0.001). Denitrification enzyme activity in the shallow A horizon of wetland soils was significantly higher (p < 0.001) than in deeper soils. Significant stimulation of DEA (p = 0.027) by N03- -N amendments occurred only in the meander scar soils receiving low NO3- -N (<3.6 mg L(-1)) concentrations. Significant denitrification of high NO3- -N ground water can occur in riparian wetland soils, but DEA is dependent upon localized differences in the degree of soil saturation and organic carbon content.


Canadian health system reforms: lessons for Australia?
This paper analyses recent health reform agenda in Canada. From 1988 until 1997, the first phase of reforms focused on service integration through regionalisation and a rebalancing of services from illness care to prevention and wellness. The second phase, which has been layered onto the ongoing first phase, is concerned with fiscal sustainability from a provincial perspective, and the fundamental nature of the system from a national perspective. Despite numerous commissions and studies, some questions remain concerning the future direction of the public system. The Canadian reform experience is compared with recent Australian health reform initiatives in terms of service integration through regionalisation, primary care reform, Aboriginal health, the public-private debate, intergovernmental relations and the role of the federal government.


Nocturnal "humming" vocalizations: adding a piece to the puzzle of giraffe vocal communication.
Recent research reveals that giraffes (Giraffa camelopardalis sp.) exhibit a socially structured, fission-fusion system. In other species possessing this kind of society, information exchange is important and vocal communication is usually well developed. But is this true for giraffes? Giraffes are known to produce sounds, but there is no evidence that they use vocalizations for communication. Reports on giraffe vocalizations are mainly anecdotal and the missing acoustic descriptions make it difficult to establish a call nomenclature. Despite inconclusive evidence to date, it is widely assumed that giraffes produce infrasonic vocalizations similar to elephants. In order to initiate a more detailed investigation of the vocal communication in giraffes, we collected data of captive individuals during day and night. We particularly focussed on detecting tonal, infrasonic or sustained vocalizations. We collected over 947 h of audio material in three European zoos and quantified the spectral and temporal components of acoustic signals to obtain an accurate set of acoustic parameters. Besides the known burst, snorts and grunts, we detected harmonic, sustained and frequency-modulated "humming" vocalizations during night recordings. None of the recorded vocalizations were within the infrasonic range. These results show that giraffes do produce vocalizations, which, based on their acoustic structure, might have the potential to function as communicative signals to convey information about the physical and motivational attributes of the caller. The data further reveal that the assumption of infrasonic communication in giraffes needs to be considered with caution and requires further investigations in future studies.


Enzymatic oxidation of cholesterol: properties and functional effects of cholestenone in cell membranes.
Bacterial cholesterol oxidase is commonly used as an experimental tool to reduce cellular cholesterol content. That the treatment also generates the poorly degradable metabolite 4-cholesten-3-one (cholestenone) has received less attention. Here, we investigated the membrane partitioning of cholestenone using simulations and cell biological experiments and assessed the functional effects of cholestenone in human cells. Atomistic simulations predicted that cholestenone reduces membrane order, undergoes faster flip-flop and desorbs more readily from membranes than cholesterol. In primary human fibroblasts, cholestenone was released from membranes to physiological extracellular acceptors more avidly than cholesterol, but without acceptors it remained in cells over a day. To address the functional effects of cholestenone, we studied fibroblast migration during wound healing. When cells were either cholesterol oxidase treated or part of cellular cholesterol was exchanged for cholestenone with cyclodextrin, cell migration during 22 h was markedly inhibited. Instead, when a similar fraction of cholesterol was removed using cyclodextrin, cells replenished their cholesterol content in 3 h and migrated similarly to control cells. Thus, cholesterol oxidation produces long-term functional effects in cells and these are in part due to the generated membrane active cholestenone.


Uptake of HIV testing among adolescents and associated adolescent-friendly services.
HIV testing remains low among adolescents. Making public health services more adolescent-friendly is one strategy used to encourage testing. However, it remains unclear whether government-led initiatives have a meaningfully impact. The current study is observational and utilizes two sources of data (health-facility and adolescent-level) from one round of data collection of an on-going, longitudinal impact evaluation of a pilot cash plus program targeting adolescents. This study linked data from adolescent surveys (n = 2191) to data collected from nearby government-run health facilities (n = 91) in two rural regions of Tanzania. We used log binomial regression models to estimate the association between specific adolescent-friendly health service (AFHS) characteristics and adolescents' uptake of 1) HIV testing and 2) visiting a health care facility in the past year for sexual and reproductive health (SRH) services. Most adolescents (67%) lived in a village with a health facility, and all offered HIV services. We find, however, that AFHS have not been fully implemented. For example, less than 40% of facilities reported that they had guidelines for adolescent care. Only 12% of facilities had a system in place for referral and follow-up with adolescent clients, yet this was an important predictor of both past-year HIV testing (RR = 1.28, p < 0.1) and SRH visits (RR = 1.44, p < 0.05). Less than half (44%) offered services for survivors of gender-based violence (GBV), a significant predictor of past-year HIV testing (RR = 1.20, p < 0.05) and SRH visits (RR = 1.41, p < 0.01) among sexually-active adolescents. We find that national guidelines on AFHS have not been fully translated into practice at the local level. We highlight particular gaps in adolescent referral systems and GBV services. Scaling up these two essential services could encourage greater HIV testing among a high-risk population, in addition to providing much needed support for survivors of violence.


Correlation of Severity of Fuchs Endothelial Corneal Dystrophy With Triplet Repeat Expansion in TCF4.
The CTG18.1 triplet repeat expansion in TCF4 has recently been found to be a common functional variant contributing significant risk to the development of Fuchs endothelial corneal dystrophy (FECD) in Eurasian populations. To determine the effect of the expanded CTG18.1 allele of TCF4 on FECD severity and to correlate CTG triplet repeat allele length to the severity of FECD. In a cross-sectional analysis, we studied 139 index cases (probands and unrelated individuals) with FECD recruited from a cornea referral practice at the University of Texas Southwestern Medical Center, Dallas, from April 2010 through February 2015. The triplet repeat polymorphism CTG18.1 was genotyped using a combination of short tandem repeat analysis, triplet repeat-primed polymerase chain reaction assay, and Southern blot analysis. Severity of FECD was graded using a modified Krachmer grading system (severity scale of 0-6 based on extent of confluent guttae). The CTG triplet repeat length of the largest allele was compared with the Krachmer grade of FECD severity, keratoplasty proportion, and central corneal thickness in the white subset. Eighty-five of 122 white index cases with FECD (69.7%) harbored the triplet repeat expansion. The mean (SD) Krachmer grade was 5.61 (0.76) in the group with the repeat expansion compared with 5.11 (1.05) in the group without the expanded repeats (P = .01). Forty-seven participants with the repeat expansion (55.3%) had undergone keratoplasty at the time of recruitment, compared with 13 (35.1%) of those without the expansion (P = .0497). There was a positive correlation of Krachmer grade to triplet repeat number (P = .002) and a nominal association of the keratoplasty proportion with triplet repeat number (P = .04). The mean (SD) central corneal thickness was 605.9 (50.5) μm in the group with the expanded repeats compared with 581.3 (50.5) μm in the group without the expansion (P = .04). The Krachmer grade of disease severity was greater in FECD cases with the CTG18.1 triplet repeat expansion in TCF4 than in those without the expansion. The CTG triplet repeat allele length was positively correlated with the Krachmer grade of severity. The TCF4 triplet repeat expansion resulted in a more severe form of FECD, with clinical and surgical therapeutic implications.


Diet and exercise in low-income culturally diverse middle school students.
It is important to understand constructs essential to low-fat-diet and physical-activity behaviors of low-income culturally diverse middle-school-age students, because ethnicity and socioeconomic status influence body weight status, and these groups are most at risk for obesity later in life. In this descriptive study, constructs important in low-fat diet and physical activity in low- to middle-income, culturally diverse middle school students were examined. Revised for cultural/developmental appropriateness and reading level, instruments incorporating constructs from the Health Promotion and Transtheoretical Models were tested with 221 youth. Results demonstrated that percentage of fat in diet, total number of strategies used for a low-fat diet, access to low-fat food, and total number of exercise processes varied significantly (p < 0.05) across stages of change for low-fat diet. We concluded that interventions should foster access to low-fat foods and processes for diet and activity change. Despite significantly lower income and higher grade level (both of which are risks for poor health behaviors), students in the private school demonstrated significantly lower fat in diet, higher perception of benefits, and better access to low-fat food. These findings warrant additional study.


Ultrastructural evidence for hair cell regeneration in the mammalian inner ear.
It has long been thought that hair cell loss from the inner ears of mammals is irreversible. This report presents scanning electron micrographs and thin sections of the utricles from the inner ears of guinea pigs that show that, after hair cell loss caused by treatment with the aminoglycoside gentamicin, hair cells reappeared. Four weeks after the end of treatment, a large number of cells with immature hair bundles in multiple stages of development could be identified in the utricle. Thin sections showed that lost type 1 hair cells were replaced by cells with a morphology similar to that of type 2 hair cells. These results indicate an unexpected capacity for hair cell regeneration in vivo in the mature mammalian inner ear.


Detection of Legionella spp. in bronchoalveolar lavage fluids by DNA amplification.
By using Taq polymerase, DNA amplification of a specific fragment of the macrophage infectivity potentiator (mip) gene from Legionella pneumophila was used to detect Legionella spp. in bronchoalveolar lavage (BAL) fluid specimens. We were able to detect DNAs from all 30 L. pneumophila strains tested (serogroups 1 to 14), L. micdadei, and L. bozemanii serogroup 1. DNA from bacteria of other species tested and DNA from human leukocytes were not amplified by this procedure. After optimization of the conditions for DNA extraction from BAL fluid, a 2-ml sample of BAL fluid seeded with 25 CFU/ml tested positive after DNA amplification. A total of 68 frozen BAL fluid specimens sent to the laboratory because of suspected legionellosis were tested in a retrospective study. The eight culture-positive samples were all positive after specific DNA amplification. Among 60 culture-negative samples, 7 were positive after amplification. Of these seven samples, four were from patients who had presented a typical clinical history of legionellosis; the samples had antibody titer increases of 2 dilutions. For the three remaining samples, serological diagnosis of legionellosis in the patients from whom the samples were obtained could not be documented, and although the causative agent of these pulmonary infections was not determined, the clinical features of the patients were in accordance with legionellosis.


The prevalence of statistical reporting errors in psychology (1985-2013).
This study documents reporting errors in a sample of over 250,000 p-values reported in eight major psychology journals from 1985 until 2013, using the new R package "statcheck." statcheck retrieved null-hypothesis significance testing (NHST) results from over half of the articles from this period. In line with earlier research, we found that half of all published psychology papers that use NHST contained at least one p-value that was inconsistent with its test statistic and degrees of freedom. One in eight papers contained a grossly inconsistent p-value that may have affected the statistical conclusion. In contrast to earlier findings, we found that the average prevalence of inconsistent p-values has been stable over the years or has declined. The prevalence of gross inconsistencies was higher in p-values reported as significant than in p-values reported as nonsignificant. This could indicate a systematic bias in favor of significant results. Possible solutions for the high prevalence of reporting inconsistencies could be to encourage sharing data, to let co-authors check results in a so-called "co-pilot model," and to use statcheck to flag possible inconsistencies in one's own manuscript or during the review process.


Protection against experimental autoimmune encephalomyelitis generated by a recombinant adenovirus vector expressing the V beta 8.2 TCR is disrupted by coadministration with vectors expressing either IL-4 or -10.
Adenovirus vectors are increasingly being used for genetic vaccination and may prove highly suitable for intervention in different pathological conditions due to their capacity to generate high level, transient gene expression. In this study, we report the use of a recombinant adenovirus vector to induce regulatory responses for the prevention of autoimmune diseases through transient expression of a TCR beta-chain. Immunization of B10.PL mice with a recombinant adenovirus expressing the TCR Vbeta8.2 chain (Ad5E1 mVbeta8.2), resulted in induction of regulatory type 1 CD4 T cells, directed against the framework region 3 determinant within the B5 peptide (aa 76-101) of the Vbeta8.2 chain. This determinant is readily processed and displayed in an I-A(u) context, on ambient APC. Transient genetic delivery of the TCR Vbeta8.2 chain protected mice from Ag-induced experimental autoimmune encephalomyelitis. However, when the Ad5E1 mVbeta8.2 vector was coadministered with either an IL-4- or IL-10-expressing vector, regulation was disrupted and disease was exacerbated. These results highlight the importance of the Th1-like cytokine requirement necessary for the generation and activity of effective regulatory T cells in this model of experimental autoimmune encephalomyelitis.


Seeking common ground: a history of labor and Blue Cross.
In recent years, voluntary health insurance costs have become a major source of friction in labor-management negotiations. What was once a "fringe" has led to job actions, strikes, and intensive bargaining. We examine the history of labor's participation in New York Blue Cross from the 1930s to the recent past and show that labor's participation in the plan was crucial to Blue Cross's success in the plan's early decades. By the late 1950s, serious tensions developed over rate increases and the participation of labor in Blue Cross governance. Ultimately, the issue was one of the control over what was provided by the plans and who would pay for the costs of care. We posit that labor was never able to achieve an important role in the control of the third-party payer, and in the antilabor environment of the 1980s this proved detrimental to labor's interests.


CRISPR-Mediated Single Nucleotide Polymorphism Modeling in Rats Reveals Insight Into Reduced Cardiovascular Risk Associated With Mediterranean G6PD Variant.
Epidemiological studies suggest that individuals in the Mediterranean region with a loss-of-function, nonsynonymous single nucleotide polymorphism (S188F), in glucose-6-phosphate dehydrogenase (G6pd) are less susceptible to vascular diseases. However, this association has not yet been experimentally proven. Here, we set out to determine whether the Mediterranean mutation confers protection from vascular diseases and to discover the underlying protective mechanism. We generated a rat model with the Mediterranean single nucleotide polymorphism (G6PDS188F) using CRISPR-Cas9 genome editing. In rats carrying the mutation, G6PD activity, but not expression, was reduced to 20% of wild-type (WT) littermates. Additionally, unbiased metabolomics analysis revealed that the pentose phosphate pathway and other ancillary metabolic pathways connected to the pentose phosphate pathway were reduced (P<0.05) in the arteries of G6PDS188F versus WT rats. Intriguingly, G6PDS188F mutants, as compared with WT rats, developed less large arterial stiffness and hypertension evoked by high-fat diet and nitric oxide synthase inhibition with L-NG-nitroarginine methyl ester. Intravenous injection of a voltage-gated L-type Ca2+ channel agonist (methyl 2,6-dimethyl-5-nitro-4-[2-(trifluoromethyl)phenyl]-1,4-dihydropyridine-3-carboxylate; Bay K8644) acutely increased blood pressure in WT but not in G6PDS188F rats. Finally, our results suggested that (1) lower resting membrane potential of smooth muscle caused by increased expression of K+ channel proteins and (2) decreased voltage-gated Ca2+ channel activity in smooth muscle contributed to reduced hypertension and arterial stiffness evoked by L-NG-nitroarginine methyl ester and high-fat diet to G6PDS188F mutants as compared with WT rats. In summary, a mutation resulting in the replacement of a single amino acid (S188F) in G6PD, the rate-limiting enzyme in the pentose phosphate pathway, ascribed properties to the vascular smooth muscle that shields the organism from risk factors associated with vascular diseases.


Global elongation and high shape flexibility as an evolutionary hypothesis of accommodating mammalian brains into skulls.
Little is known about how the large brains of mammals are accommodated into the dazzling diversity of their skulls. It has been suggested that brain shape is influenced by relative brain size, that it evolves or develops according to extrinsic or intrinsic mechanical constraints, and that its shape can provide insights into its proportions and function. Here, we characterize the shape variation among 84 marsupial cranial endocasts of 57 species including fossils, using three-dimensional geometric morphometrics and virtual dissections. Statistical shape analysis revealed four main patterns: over half of endocast shape variation ranges from elongate and straight to globular and inclined; little allometric variation with respect to centroid size, and none for relative volume; no association between locomotion and endocast shape; limited association between endocast shape and previously published histological cortex volumes. Fossil species tend to have smaller cerebral hemispheres. We find divergent endocast shapes in closely related species and within species, and diverse morphologies superimposed over the main variation. An evolutionarily and individually malleable brain with a fundamental tendency to arrange into a spectrum of elongate-to-globular shapes-possibly mostly independent of brain function-may explain the accommodation of brains within the enormous diversity of mammalian skull form.


Tgfβ/Alk5 signaling is required for shear stress induced klf2 expression in embryonic endothelial cells.
Endothelial cells (EC) translate biomechanical forces into functional and phenotypic responses that play important roles in cardiac development. Specifically, EC in areas of high shear stress, i.e., in the cardiac outflow tract and atrioventricular canal, are characterized by high expression of Krüppel-like factor 2 (Klf2) and by transforming growth factor-beta (Tgfβ)-driven endothelial-to-mesenchymal transition. Extraembryonic venous obstruction (venous clip model) results in congenital heart malformations, and venous clip-induced alterations in shear stress-related gene expression are suggestive for an increase in cardiac shear stress. Here, we study the effects of shear stress on Klf2 expression and Tgfβ-associated signaling in embryonic EC in vivo using the venous clip model and in vitro by subjecting cultured EC to fluid flow. Cellular responses were assessed by analysis of Klf2, Tgfβ ligands, and their downstream signaling targets. Results show that, in embryonic EC, shear stress activates Tgfβ/Alk5 signaling and that induction of Klf2 is an Alk5 dependent process.


Adjunctive everolimus for children and adolescents with treatment-refractory seizures associated with tuberous sclerosis complex: post-hoc analysis of the phase 3 EXIST-3 trial.
Epilepsy occurs in 70-90% of patients with tuberous sclerosis complex. We aimed to assess the efficacy and safety of adjunctive everolimus for treatment-refractory seizures associated with tuberous sclerosis complex in paediatric patients enrolled in the EXIST-3 trial, a double-blind, placebo-controlled, randomised, phase 3 study. This post-hoc analysis focused on paediatric patients (age <18 years) in the EXIST-3 trial, which consisted of baseline (8 weeks), core (18 weeks), and extension phases (≥48 weeks) and was done at 99 centres in 25 countries worldwide. Briefly, patients with tuberous sclerosis complex-associated treatment-refractory seizures, who were receiving a stable dose of one to three antiepileptic drugs, were randomly assigned (1:1:1) to receive placebo, low-exposure everolimus (3-7 ng/mL), or high-exposure everolimus (9-15 ng/mL). Following the core phase, patients could enter the extension phase to receive everolimus at a targeted exposure range of 3-15 ng/mL up to 48 weeks after the last patient had completed the core phase. Efficacy endpoints were response rate (≥50% of reduction from baseline in average weekly seizure frequency) and median percentage reduction in seizure frequency during the 12-week maintenance period of the core phase, and at 12-week intervals throughout the extension phase. This study is registered with ClinicalTrials.gov, number NCT01713946. Between July 3, 2013, and May 29, 2015, 299 paediatric patients enrolled in the trial. In the younger subgroup (<6 years; n=104), 34 received placebo, 33 low-exposure everolimus, and 37 high-exposure everolimus; in the older subgroup (≥6 years to <18 years; n=195), 62 received placebo, 63 low-exposure everolimus, and 70 high-exposure everolimus. At the end of the core phase, response rate was higher in the treatment groups than placebo in both the younger subgroup (17·6% [6·8-34·5] for placebo vs 30·3% [95% CI 15·6-48·7; p=0·2245] for low-exposure everolimus vs 59·5% [42·1-75·2; p=0·0003] for high-exposure everolimus) and the older subgroup (12·9% [5·7-23·9] vs 27·0% [16·6-39·7; p=0·0491] vs 30·0% [19·6-42·1; p=0·0179]), as were median reduction in seizure frequency (12·3% [95% CI -10·1 to 24·8] vs 29·3% [95% CI 13·4 to 46·3; p=0·0474] vs 54·7% [43·5 to 73·1; p<0·0001] in younger patients; 13·5% [-3·0 to 26·8] vs 31·0% [16·1 to 42·9; p=0·0128] vs 34·8% [26·7 to 41·3; p=0·0006] in older patients). The efficacy persisted, with sustained seizure reduction after 1 year of treatment across both paediatric subgroups (response rate 48·9% [95% CI 38·1-59·8] for the younger subgroup vs 47·2% [39·3-55·2] for the older subgroup; median percentage reduction in seizure frequency 48·4% [95% CI 34·3-73·6] vs 48·0% [38·2-57·5]). At the cutoff date for the extension phase, grade 3 or 4 adverse events were reported in 45 (45%) younger patients (commonly pneumonia [n=16]) and 74 (38%) older patients (commonly pneumonia [n=8] and stomatitis [n=6]). Two deaths (pneumonia, which was suspected to be treatment-related, and sudden unexplained death due to epilepsy) were reported. Adjunctive everolimus resulted in sustained reductions in seizure frequency after 1 year and was well tolerated in paediatric patients with treatment-refractory seizures associated with tuberous sclerosis complex. Novartis Pharmaceuticals Corporation.


The opposite end of the attention deficit hyperactivity disorder continuum: genetic and environmental aetiologies of extremely low ADHD traits.
Although attention deficit hyperactivity disorder (ADHD) is thought to reflect a continuously distributed quantitative trait, it is assessed through binary diagnosis or skewed measures biased towards its high, symptomatic extreme. A growing trend is to study the positive tail of normally distributed traits, a promising avenue, for example, to study high intelligence to increase power for gene-hunting for intelligence. However, the emergence of such a 'positive genetics' model has been tempered for ADHD due to poor phenotypic resolution at the low extreme. Overcoming this methodological limitation, we conduct the first study to assess the aetiologies of low extreme ADHD traits. In a population-representative sample of 2,143 twins, the Strength and Weaknesses of ADHD Symptoms and Normal behaviour (SWAN) questionnaire was used to assess ADHD traits on a continuum from low to high. Aetiological influences on extreme ADHD traits were estimated using DeFries-Fulker extremes analysis. ADHD traits were related to behavioural, cognitive and home environmental outcomes using regression. Low extreme ADHD traits were significantly influenced by shared environmental factors (23-35%) but were not significantly heritable. In contrast, high-extreme ADHD traits showed significant heritability (39-51%) but no shared environmental influences. Compared to individuals with high extreme or with average levels of ADHD traits, individuals with low extreme ADHD traits showed fewer internalizing and externalizing behaviour problems, better cognitive performance and more positive behaviours and positive home environmental outcomes. Shared environmental influences on low extreme ADHD traits may reflect passive gene-environment correlation, which arises because parents provide environments as well as passing on genes. Studying the low extreme opens new avenues to study mechanisms underlying previously neglected positive behaviours. This is different from the current deficit-based model of intervention, but congruent with a population-level approach to improving youth wellbeing.


Arenium acid catalyzed deuteration of aromatic hydrocarbons.
The arenium acid [mesitylene-H](+) has been shown to be an extraordinarily active H/D exchange catalyst for the perdeuteration of polycyclic aromatic hydrocarbons. The reactions take place under ambient conditions in C6D6 as an inexpensive deuterium source. High isolated yields and excellent degrees of deuterium incorporation were achieved using the substrates p-terphenyl, fluoranthene, pyrene, triphenylene, and corannulene.


Breast Tumor Microcalcification Induced by Bone Morphogenetic Protein-2: A New Murine Model for Human Breast Tumor Diagnosis.
Widespread use of screening mammography has recently increased the detection of breast microcalcifications. These nonpalpable microcalcifications with specific features in breast tissues are clinically considered an early indicator of breast carcinoma. Our goal in this study was to develop a murine breast microcalcification model for optimizing in vivo imaging. Recombinant human BMP-2 was expressed in E. coli, and the purified bioactive protein was used as inducing factor for the production of breast microcalcifications in a murine animal model. Syngeneic breast tumors were obtained by injection of MDA-MB-231 human breast cancer cells with Matrigel into the mammary fat pad of female nude mice. Different doses of bioactive rhBMP-2 were administered either as single or multiple intraperitoneal injections or directly into tumor on a weekly basis. Three weeks after the first injection of rhBMP-2, the microcalcification of breast tumor was detected by microcomputed tomography followed by intravenous injection of radiotracer [18F] Sodium fluoride for positron emission tomography imaging. Our findings indicate that rhBMP-2 induced microcalcifications of breast tumor by both systemic and direct injection of rhBMP-2 into tumors in a dose-dependent manner. Although little is known about the molecular mechanism of microcalcification, here we report a new murine model of human breast tumor induced microcalcification by rhBMP-2 to optimize in vivo imaging methods and to study the role of BMP-2 as a mediator of pathological mineralization and bone-like microcalcification formation in breast tumor. This BMP-2-induced microcalcification model may allow us to discriminate the type of microcalcification in tumors and to perform quantitative analysis on the calcification as a new detection strategy for early identification of pathological mineralization of breast tissues in women.


Estimating the Costs of Torture: Challenges and Opportunities.
Due to its nature, extent and consequences, torture is considered a major public health problem and a serious violation of human rights. Our study aims to set the foundation for a theoretical framework of the costs related to torture. It examines existing challenges and proposes some solutions. Our proposed framework targets policy makers, human rights activists, professionals working in programmes, centres and rehabilitation projects, judges and lawyers, survivors of torture and their families and anyone involved in the prevention and fight against this practice and its consequences. We adopted a methodology previously used in studies investigating the challenges in measuring and valuing productivity costs in health disorders. We identify and discuss conceptual, methodological, political and ethical challenges that studies on the economic and social costs of torture pose and propose alternatives in terms of possible solutions to these challenges. The economic dimension of torture is rarely debated and integrated in research, policies and programmes. Several challenges such as epistemological, methodological, ethical or political ones have often been presented as obstacles to cost studies of torture and as an excuse for not investigating this dimension. In identifying, analysing and proposing solutions to these challenges, we intend to stimulate the integration of the economic dimension in research and prevention of torture strategies.


Wild and farmed salmon (Salmo salar) as reservoirs for infectious salmon anaemia virus, and the importance of horizontal- and vertical transmission.
The infectious salmon anaemia virus (ISAV) is an important pathogen on farmed salmon in Europe. The virus occurs as low- and high virulent variants where the former seem to be a continuous source of new high virulent ISAV. The latter are controlled in Norway by stamping out infected populations while the former are spreading uncontrolled among farmed salmon. Evidence of vertical transmission has been presented, but there is still an ongoing discussion of the importance of circulation of ISAV via salmon brood fish. The only known wild reservoirs are in trout (Salmo trutta) and salmon (Salmo salar). This study provides the first ISAV sequences from wild salmonids in Norway and evaluates the importance of this reservoir with respect to outbreaks of ISA among farmed salmon. Phylogenetic analyses of the surface protein hemagglutinin-esterase gene from nearly all available ISAV from Norway, Faeroe Islands, Scotland, Chile and wild salmonids in Norway show that they group into four major clades. Including virulent variants in the analysis show that they belong in the same four clades supporting the hypothesis that there is a high frequency of transition from low to high virulent variants in farmed populations of salmon. There is little support for a hypothesis suggesting that the wild salmonids feed the virus into farmed populations. This study give support to earlier studies that have documented local horizontal transmission of high virulent ISAV, but the importance of transition from low- to high virulent variants has been underestimated. Evidence of vertical transmission and long distance spreading of ISAV via movement of embryos and smolt is presented. We recommend that the industry focus on removing the low virulent ISAV from the brood fish and that ISAV-free brood fish salmon are kept in closed containment systems (CCS).


The incidence of anemia in an Israeli population: a population analysis for anemia in 34,512 Israeli infants aged 9 to 18 months.
The purpose of this work was to use the comprehensive computerized database of Clalit Health Services to analyze the prevalence and contributing factors of anemia among the population of Clalit Health Services-insured Israeli infants aged 9 to 18 months, characterized by ethnic sector. This was a cross-sectional retrospective study for the year 2003 using the computerized database of Clalit Health Services for 34,512 infants aged 9 to 18 months insured by the Clalit Health Services sick fund. Children with abnormal white blood counts at the time of the hemoglobin test and with chronic diseases were excluded. The data were analyzed for age, infant hemoglobin level, ethnic origin, district distribution, type of clinic where the infant received treatment, the number of iron prescriptions dispensed to each child, and the mother's last hemoglobin level before giving birth. Anemia was defined as a hemoglobin level <105 g/L. The prevalence of anemia among Israeli infants is 15.5%. The prevalence is significantly higher in the non-Jewish population (22.5%) as compared with the Jewish population (10.5%). The lowest prevalence of anemia was found in pediatric health centers (10.7%). A significant correlation was found between the presence of anemia in infants and the presence of anemia found in their mothers. Infants with anemia used significantly less iron preparations. This study is one of the first studies to use a comprehensive computerized database to perform a population-based analysis of anemic infants. We found a considerable percentage of infants to be anemic and identified a specific population to be at high risk for anemia. We describe 2 factors that have the potential to be altered through intervention: improving compliance of iron intake and maternal anemia. Major national efforts should be made to minimize the prevalence of anemia, especially in the non-Jewish population, and to learn more about the causes of iron-deficiency anemia in this group. This study provides a base for an intervention study.