Usefulness of skin test in Farmer's lung.
The skin test with hay extract or with Micropolyspora faeni (MF), is not commonly used in the diagnosis of Farmer's lung (FL), as it is not considered specific. In our study, we have applied the intracutaneous test with these antigens in 26 patients affected with FL; 18 of them were still in contact with the antigen and the remaining eight had not been in contact with the hay during the previous year. Twenty-five asymptomatic farmers (AF) served as a control group. In the first group and with hay extract, the immediate reading (I) was positive in 15 of 18 (83.3 percent), the late reaction (L) in 18 of 18 (100 percent), and the delayed one (D) in eight of 18 (44.4 percent). In the 25 AF, the results were as follows: I, seven of 85 (28 percent); L, 17 of 25 (68 percent); and D, one of 25 (4 percent). Consequently, the differences between both groups were significant: I, p less than 0.01; L, p less than 0.05; and D, p less than 0.01. Using MF as an antigen, the test is somewhat less effective: p less than 0.02, p less than 0.02, and p less than 0.2, respectively. These results suggest that the intradermal injection with hay extract is an easy, effective test in the diagnosis of FL, and at the same time, a better means of distinguishing FL patients from AF than the precipitation test.


Graphene materials as 2D non-viral gene transfer vector platforms.
Advances in genomics and gene therapy could offer solutions to many diseases that remain incurable today, however, one of the critical reasons halting clinical progress is due to the difficulty in designing efficient and safe delivery vectors for the appropriate genetic cargo. Safety and large-scale production concerns counter-balance the high gene transfer efficiency achieved with viral vectors, while non-viral strategies have yet to become sufficiently efficient. The extraordinary physicochemical, optical and photothermal properties of graphene-based materials (GBMs) could offer two-dimensional components for the design of nucleic acid carrier systems. We discuss here such properties and their implications for the optimization of gene delivery. While the design of such vectors is still in its infancy, we provide here an exhaustive and up-to-date analysis of the studies that have explored GBMs as gene transfer vectors, focusing on the functionalization strategies followed to improve vector performance and on the biological effects attained.


Auditory Brainstem Responses (ABR) changes in children treated with high doses cisplatin.
Cisplatin can cause cochlear damage by oxidative stress in hair cells, but there are few studies regarding toxicity in the central auditory pathways. To study cisplatin neurotoxicity in the central auditory pathways in children treated with high doses cisplatin for solid malignant tumors. Thirteen children (Group S) aged 3-19 years who had been treated with cisplatin (60-120 mg/m(2)/cycle) were studied using evoked otoacoustic emissions (EOA), conventional auditory evaluation and auditory brainstem response (ABR). Thirteen normal children matched for age and sex composed the control group (Group C). Group S - 7.5% percent of ears showed enlargement of latencies of waves I and V, and 3.8% of wave III. Six ears (23%) showed enlarged interpeak intervals. Four ears showed increased interpeak I-III and two showed increased interpeak III-V. The results were compared using the Mann-Whitney test. Interpeak III-V differed significantly between groups S and C when only the left ears were considered. EOA results were normal in both groups. Abnormal values in the interpeak I-III associated with normal distortion product OEA suggest neurotoxicity in the brainstem pathways. The statistical significance reached only in the left ear may be due to small number of cases studied.


Study of the flow field in the magnetic rod interfacial stress rheometer.
Several technological applications, consumer products, and biological systems derive their functioning from the presence of a complex fluid interface with viscoelastic interfacial rheological properties. Measurements of the "excess" rheological properties of such an interface are complicated by the intimate coupling of the bulk and interfacial flows. In the present work, analytical, numerical, and experimental results of the interfacial flow fields in a magnetic rod interfacial stress rheometer (ISR) are presented. Mathematical solutions are required to correct the experimentally determined apparent interfacial shear moduli and phase angles for the drag exerted by the surrounding phases, especially at low Boussinesq numbers. Starting from the Navier-Stokes equations and using the generalized Boussinesq-Scriven equation as a suitable boundary condition, the problem is solved both analytically and numerically. In addition, experimental data of the interfacial flow field are reported, obtained by following the trajectories of tracer particles at the interface with time. Good agreement is found between the three methods, indicating that both the analytical solution and the numerical simulations give an adequate description of the flow field and the resulting local interfacial shear rate at the rod. Based on these results, an algorithm to correct the experimental data of the ISR is proposed and evaluated, which can be extended to different types of interfacial shear rheometers and geometries. An increased accuracy is obtained and the measurement range of the ISR is expanded toward viscosities and elastic moduli of smaller magnitude.


Standardized patients' perceptions about their own health care.
There have been detailed descriptions on standardized patients (SP) programs' effects on students, curricula, and faculty, yet little attention has been paid to the consequences of participating on the SP's. This study explored the perceptions of SPs toward their own health care in the context of having served as SPs. All 180 SPs participating in Department of Medicine programs at 5 medical schools were surveyed. They completed the survey during SP activities, or it was mailed to them. SPs indicated their level of agreement or disagreement with 11 attitude statements related to their own health care after serving as an SP using a Likert scale, with 1 reflecting the most positive attitude and 5 the least positive. Responses to the attitudinal questions were obtained from 164 SPs (91%). SPs perceived that because of their participation as SPs they had a better understanding about medical history taking and physical examinations (1.9 +/- 0.9), communicated more effectively with their health care provider (1.8 +/- 0.9), and were more comfortable with both health care visits and physical examinations (2.2 +/- 0.9). There were no significant differences in results based on gender, age, race, or school. As a consequence of their participation, the SPs indicated a change in attitudes about their personal health care. They perceived improved understanding and ability to communicate and comfort with their own health care. Participation in SP programs seems to influence SPs by improving perceptions about their own health care interactions.


Suicidal behaviour and the menstrual cycle.
To investigate the available data regarding possible associations between phases of the menstrual cycle and suicidal behaviour. A systematic search of major relevant databases was performed using appropriate search terms. Forty-four relevant studies were identified in total, 13 pertaining to suicide, two pertaining to both attempted and completed suicide, 23 to suicide attempts, three to suicide attempts in those suffering from the premenstrual syndrome (PMS), three to suicidal ideation and two to repetitive self-harming behaviours. A variety of methodologies were used in these studies and there were notable differences in the conclusions reached. Studies with better methodology suggest that a positive relationship exists between aspects of the menstrual cycle and non-fatal suicidal behaviour. Such behaviour appears to be more common in those phases of the menstrual cycle when oestrogen levels are lowest (the late luteal and follicular phases), and in those suffering from PMS. There appears to be an association between the menstrual cycle and non-fatal suicidal behaviours. Interaction between oestrogen and the serotonergic system may provide a possible mechanism.


A phase conversion method to anchor ZIF-8 onto a PAN nanofiber surface for CO2 capture.
Polyacrylonitrile (PAN) nanofibers were prepared by electrospinning and coated with zeolitic imidazolate framework-8 (ZIF-8) by a phase conversion growth method and investigated for CO2 capture. The PAN nanofibers were pre-treated with NaOH, and further coated with zinc hydroxide, which was subsequently converted into ZIF-8 by the addition of 2-methyl imidazolate. In the resulting flexible ZIF-8/PAN composite nanofibers, ZIF-8 loadings of up to 57 wt% were achieved. Scanning electron microscopy and energy-dispersive X-ray spectroscopy (EDS) showed the formation of evenly distributed submicron-sized ZIF-8 crystals on the surface of the PAN nanofibers with sizes between 20 and 75 nm. X-ray photoelectron spectroscopy (XPS) and carbon-13 nuclear magnetic resonance (13C NMR) investigations indicated electrostatic interactions and hydrogen bonds between the ZIF-8 structure and the PAN nanofiber. The ZIF-8/composite nanofibers showed a high BET surface area of 887 m2 g-1. CO2 adsorption isotherms of the ZIF-8/PAN composites revealed gravimetric CO2 uptake capacities of 130 mg g-1 (at 298 K and 40 bar) of the ZIF-8/PAN nanofiber and stable cyclic adsorption performance.


Comparison of directly measured arterial blood pressure at various anatomic locations in anesthetized dogs.
To determine whether directly measured arterial blood pressure differs among anatomic locations and whether arterial blood pressure is influenced by body position. 33 client-owned dogs undergoing anesthesia. Dogs undergoing anesthetic procedures had 20-gauge catheters placed in both the superficial palmar arch and the contralateral dorsal pedal artery (group 1 [n = 20]) or the superficial palmar arch and median sacral artery (group 2 [13]). Dogs were positioned in dorsal recumbency, and mean arterial blood pressure (MAP), systolic arterial blood pressure (SAP), and diastolic arterial blood pressure (DAP) were recorded for both arteries 4 times (2-minute interval between successive measurements). Dogs were positioned in right lateral recumbency, and blood pressure measurements were repeated. Differences were detected between pressures measured at the 2 arterial sites in both groups. This was especially true for SAP measurements in group 1, in which hind limb measurements were a mean of 16.12 mm Hg higher than carpus measurements when dogs were in dorsal recumbency and 14.70 mm Hg higher than carpus measurements when dogs were in lateral recumbency. Also, there was significant dispersion about the mean for all SAP, DAP, and MAP measurements. Results suggested that arterial blood pressures may be dependent on anatomic location and body position. Because this may affect outcomes of studies conducted to validate indirect blood pressure measurement systems, care must be used when developing future studies or interpreting previous results.


Ambient endotoxin concentrations in PM10 from Southern California.
Concentrations of endotoxin in urban air pollution have not previously been extensively characterized. We measured 24-hr levels of PM10 (particulate matter < 10 microm in aerodynamic diameter) and the associated endotoxin component once every 6 weeks for 1 year in 13 communities in Southern California. All the samples collected had detectable PM10 and endotoxin levels. The geometric mean PM10 was 34.6 microg/m3 [geometric SD (GSD), 2.1; range, 3.0-135]. By volume, the endotoxin geometric mean was 0.44 endotoxin units (EU)/m3 (GSD, 3.1; range, 0.03-5.44). Per unit material collected, the geometric mean of endotoxin collected was 13.6 EU/mg (GSD, 3.2; range, 0.7-96.8). No correlation was found between endotoxin concentrations and other ambient pollutants concurrently measured [ozone, nitrogen dioxide, total acids, or PM2.5 (particulate matter < 2.5 micro m in aerodynamic diameter]. PM10 and endotoxin concentrations were significantly correlated, most strongly in summer. Samples collected in more rural and agricultural areas had lower PM10 and mid-range endotoxin levels. The high desert and mountain communities had lower PM10 levels but endotoxin levels comparable with or higher than the rural agricultural sites. By volume, endotoxin levels were highest at sites downwind of Los Angeles, California, which were also the locations of highest PM10. Endotoxin concentrations measured in this study were all < 5.5 EU/m3, which is lower than recognized thresholds for acute adverse health effects for occupational exposures but in the same range as indoor household concentrations. This study provides the first extensive characterization of endotoxin concentration across a large metropolitan area in relation to PM10 and other pollutant monitoring, and supports the need for studies of the role of endotoxin in childhood asthma in urban settings.


Protective effect of carbon monoxide inhalation for cold-preserved small intestinal grafts.
Heme oxygenase (HO)-1 system has been shown to provide protection against oxidative stress through the degradation of heme to biliverdin, free iron, and carbon monoxide (CO). This study investigated cytoprotective efficacy of CO at a low concentration on cold ischemia/reperfusion (I/R) injury of transplanted intestine. Lewis rat recipients of syngenic orthotopic small intestinal transplantation with 6 hours UW cold preservation were either kept in room air (air-treated control) or exposed to CO (250 ppm) for 1 hour before and 24 hours after surgery. In air-treated grafts, mRNA levels for interleukin-6, intracellular adhesion molecule-1, cyclooxygenase-2, and inducible nitric oxide synthase promptly increased. Sequential histopathologic analysis of untreated grafts revealed initial rapid epithelial loss, subsequent recruitment of inflammatory infiltrates, and local hemorrhage in the lamina propria, which extended downward to the epithelial crypt and muscle layer with time. CO effectively blocked proinflammatory cascade during I/R injury, inhibited upregulation of inflammatory molecules and ameliorated intestinal tissue injuries. Beneficial effects of CO were associated with improved graft blood flow without inhibiting endogenous HO-1 activity. Recipient animal survival was significantly improved with CO to 100% versus 58% in air-treated controls. These results indicate a significant role for CO in protecting the intestine from cold I/R injury associating with small intestinal transplantation.


Targeting LOXL2 for cardiac interstitial fibrosis and heart failure treatment.
Interstitial fibrosis plays a key role in the development and progression of heart failure. Here, we show that an enzyme that crosslinks collagen-Lysyl oxidase-like 2 (Loxl2)-is essential for interstitial fibrosis and mechanical dysfunction of pathologically stressed hearts. In mice, cardiac stress activates fibroblasts to express and secrete Loxl2 into the interstitium, triggering fibrosis, systolic and diastolic dysfunction of stressed hearts. Antibody-mediated inhibition or genetic disruption of Loxl2 greatly reduces stress-induced cardiac fibrosis and chamber dilatation, improving systolic and diastolic functions. Loxl2 stimulates cardiac fibroblasts through PI3K/AKT to produce TGF-β2, promoting fibroblast-to-myofibroblast transformation; Loxl2 also acts downstream of TGF-β2 to stimulate myofibroblast migration. In diseased human hearts, LOXL2 is upregulated in cardiac interstitium; its levels correlate with collagen crosslinking and cardiac dysfunction. LOXL2 is also elevated in the serum of heart failure (HF) patients, correlating with other HF biomarkers, suggesting a conserved LOXL2-mediated mechanism of human HF.


Nonstationary cluster-size inference with random field and permutation methods.
Because of their increased sensitivity to spatially extended signals, cluster-size tests are widely used to detect changes and activations in brain images. However, when images are nonstationary, the cluster-size distribution varies depending on local smoothness. Clusters tend to be large in smooth regions, resulting in increased false positives, while in rough regions, clusters tend to be small, resulting in decreased sensitivity. Worsley et al. proposed a random field theory (RFT) method that adjusts cluster sizes according to local roughness of images [Worsley, K.J., 2002. Nonstationary FWHM and its effect on statistical inference of fMRI data. Presented at the 8th International Conference on Functional Mapping of the Human Brain, June 2-6, 2002, Sendai, Japan. Available on CD-ROM in NeuroImage 16 (2) 779-780; Hum. Brain Mapp. 8 (1999) 98]. In this paper, we implement this method in a permutation test framework, which requires very few assumptions, is known to be exact [J. Cereb. Blood Flow Metab. 16 (1996) 7] and is robust [NeuroImage 20 (2003) 2343]. We compared our method to stationary permutation, stationary RFT, and nonstationary RFT methods. Using simulated data, we found that our permutation test performs well under any setting examined, whereas the nonstationary RFT test performs well only for smooth images under high df. We also found that the stationary RFT test becomes anticonservative under nonstationarity, while both nonstationary RFT and permutation tests remain valid under nonstationarity. On a real PET data set we found that, though the nonstationary tests have reduced sensitivity due to smoothness estimation variability, these tests have better sensitivity for clusters in rough regions compared to stationary cluster-size tests. We include a detailed and consolidated description of Worsley nonstationary RFT cluster-size test.


Evaluation of the stability of fluoxetine in pluronic lecithin organogel and the determination of an appropriate beyond-use date.
Fluoxetine is a commonly prescribed psychotropic medication for a variety of behavioral diagnoses in veterinary practice, and fluoxetine in Pluronic lecithin organogel has been used successfully in treating inappropriate urine spraying in felines. Historically, pharmacists have assigned a variety of beyond-use dates to extemporaneously compound drugs in Pluronic lecithin organogel. The objective of this study was to evaluate the stability of fluoxetine in Pluronic lecithin organogel over a period of six months and to determine an appropriate beyond-use date. A stability-indicating high-performance liquid chromatography method for fluoxetine in Pluronic lecithin organogel was validated in our laboratory. Fluoxetine-Pluronic lecithin organogel 50 mg/mL was prepared by a local compounding pharmacy and analyzed by high-performance liquid chromatograph at 0, 7, 14, 21, 28, 45, 60, 90, and 180 days. Physical stability was also assessed by visual observation. At each time point percent of initial concentration was calculated. The beyond-use date was determined as the time period that the samples maintained at least 90 percent of the initial concentration. At 180 days, the mean percent of initial concentration was 99 +/- 1.5 and, visually, the fluoxetine-Pluronic lecithin organogel retained the original color and consistency, without detectable separation of the different phases of Pluronic lecithin organogel. Since fluoxetine was physically stable and retained greater than 90 percent of initial concentration in Pluronic lecithin organogel for 180 days when stored at room temperature and protected from light, a beyond-use date of 180 days is appropriate.


The second version of the Copenhagen Psychosocial Questionnaire.
The aim of the present paper is to present the development of the second version of the Copenhagen Psychosocial Questionnaire (COPSOQ II). The development of COPSOQ II took place in five main steps: (1) We considered practical experience from the use of COPSOQ I, in particular feedback from workplace studies where the questionnaire had been used; (2) All scales concerning workplace factors in COPSOQ I were analyzed for differential item functioning (DIF) with regard to gender, age and occupational status; (3) A test version of COPSOQ II including new scales and items was developed and tested in a representative sample of working Danes between 20 and 59 years of age. In all, 3,517 Danish employees participated in the study. The overall response rate was 60.4%; (4) Based on psychometric analyses, the final questionnaire was developed; and (5) Criteria-related validity of the new scales was tested. The development of COPSOQ II resulted in a questionnaire with 41 scales and 127 items. New scales on values at the workplace were introduced including scales on Trust, Justice and Social inclusiveness. Scales on Variation, Work pace, Recognition, Work-family conflicts and items on offensive behaviour were also added. New scales regarding health symptoms included: Burnout, Stress, Sleeping troubles and Depressive symptoms. In general, the new scales showed good criteria validity. All in all, 57% of the items of COPSOQ I were retained in COPSOQ II. The COPSOQ I concept has been further developed and new validated scales have been included.


LmrR: A Privileged Scaffold for Artificial Metalloenzymes.
The biotechnological revolution has made it possible to create enzymes for many reactions by directed evolution. However, because of the immense number of possibilities, the availability of enzymes that possess a basal level of the desired catalytic activity is a prerequisite for success. For new-to-nature reactions, artificial metalloenzymes (ARMs), which are rationally designed hybrids of proteins and catalytically active transition-metal complexes, can be such a starting point. This Account details our efforts toward the creation of ARMs for the catalysis of new-to-nature reactions. Key to our approach is the notion that the binding of substrates, that is, effective molarity, is a key component to achieving large accelerations in catalysis. For this reason, our designs are based on the multidrug resistance regulator LmrR, a dimeric transcription factor with a large, hydrophobic binding pocket at its dimer interface. In this pocket, there are two tryptophan moieties, which are important for promiscuous binding of planar hydrophobic conjugated compounds by π-stacking. The catalytic machinery is introduced either by the covalent linkage of a catalytically active metal complex or via the ligand or supramolecular assembly, taking advantage of the two central tryptophan moieties for noncovalent binding of transition-metal complexes. Designs based on the chemical modification of LmrR were successful in catalysis, but this approach proved too laborious to be practical. Therefore, expanded genetic code methodologies were used to introduce metal binding unnatural amino acids during LmrR biosynthesis in vivo. These ARMs have been successfully applied in Cu(II) catalyzed Friedel-Crafts alkylation of indoles. The extension to MDRs from the TetR family resulted in ARMs capable of providing the opposite enantiomer of the Friedel-Crafts product. We have employed a computationally assisted redesign of these ARMs to create a more active and selective artificial hydratase, introducing a glutamate as a general base at a judicious position so it can activate and direct the incoming water nucleophile. A supramolecularly assembled ARM from LmrR and copper(II)-phenanthroline was successful in Friedel-Crafts alkylation reactions, giving rise to up to 94% ee. Also, hemin was bound, resulting in an artificial heme enzyme for enantioselective cyclopropanation reactions. The importance of structural dynamics of LmrR was suggested by computational studies, which showed that the pore can open up to allow access of substrates to the catalytic iron center, which, according to the crystal structure, is deeply buried inside the protein. Finally, the assembly approaches were combined to introduce both a catalytic and a regulatory domain, resulting in an ARM that was specifically activated in the presence of Fe(II) salts but not Zn(II) salts. Our work demonstrates that LmrR is a privileged scaffold for ARM design: It allows for multiple assembly methods and even combinations of these, it can be applied in a variety of different catalytic reactions, and it shows significant structural dynamics that contribute to achieving the desired catalytic activity. Moreover, both the creation via expanded genetic code methods as well as the supramolecular assembly make LmrR-based ARMs highly suitable for achieving the ultimate goal of the integration of ARMs in biosynthetic pathways in vivo to create a hybrid metabolism.


Heterotrimerization of heat-shock factors 1 and 2 provides a transcriptional switch in response to distinct stimuli.
Organisms respond to circumstances threatening the cellular protein homeostasis by activation of heat-shock transcription factors (HSFs), which play important roles in stress resistance, development, and longevity. Of the four HSFs in vertebrates (HSF1-4), HSF1 is activated by stress, whereas HSF2 lacks intrinsic stress responsiveness. The mechanism by which HSF2 is recruited to stress-inducible promoters and how HSF2 is activated is not known. However, changes in the HSF2 expression occur, coinciding with the functions of HSF2 in development. Here, we demonstrate that HSF1 and HSF2 form heterotrimers when bound to satellite III DNA in nuclear stress bodies, subnuclear structures in which HSF1 induces transcription. By depleting HSF2, we show that HSF1-HSF2 heterotrimerization is a mechanism regulating transcription. Upon stress, HSF2 DNA binding is HSF1 dependent. Intriguingly, when the elevated expression of HSF2 during development is mimicked, HSF2 binds to DNA and becomes transcriptionally competent. HSF2 activation leads to activation of also HSF1, revealing a functional interdependency that is mediated through the conserved trimerization domains of these factors. We propose that heterotrimerization of HSF1 and HSF2 integrates transcriptional activation in response to distinct stress and developmental stimuli.


Discontinuous shear thickening of frictional hard-sphere suspensions.
Discontinuous shear thickening (DST) observed in many dense athermal suspensions has proven difficult to understand and to reproduce by numerical simulation. By introducing a numerical scheme including both relevant hydrodynamic interactions and granularlike contacts, we show that contact friction is essential for having DST. Above a critical volume fraction, we observe the existence of two states: a low viscosity, contactless (hence, frictionless) state, and a high viscosity frictional shear jammed state. These two states are separated by a critical shear stress, associated with a critical shear rate where DST occurs. The shear jammed state is reminiscent of the jamming phase of granular matter. Continuous shear thickening is seen as a lower volume fraction vestige of the jamming transition.


The physiological period length of the human circadian clock in vivo is directly proportional to period in human fibroblasts.
Diurnal behavior in humans is governed by the period length of a circadian clock in the suprachiasmatic nuclei of the brain hypothalamus. Nevertheless, the cell-intrinsic mechanism of this clock is present in most cells of the body. We have shown previously that for individuals of extreme chronotype ("larks" and "owls"), clock properties measured in human fibroblasts correlated with extreme diurnal behavior. In this study, we have measured circadian period in human primary fibroblasts taken from normal individuals and, for the first time, compared it directly with physiological period measured in vivo in the same subjects. Human physiological period length was estimated via the secretion pattern of the hormone melatonin in two different groups of sighted subjects and one group of totally blind subjects, each using different methods. Fibroblast period length was measured via cyclical expression of a lentivirally delivered circadian reporter. Within each group, a positive linear correlation was observed between circadian period length in physiology and in fibroblast gene expression. Interestingly, although blind individuals showed on average the same fibroblast clock properties as sighted ones, their physiological periods were significantly longer. We conclude that the period of human circadian behaviour is mostly driven by cellular clock properties in normal individuals and can be approximated by measurement in peripheral cells such as fibroblasts. Based upon differences among sighted and blind subjects, we also speculate that period can be modified by prolonged unusual conditions such as the total light deprivation of blindness.


Twenty years of West Nile virus spread and evolution in the Americas visualized by Nextstrain.
It has been 20 years since West Nile virus first emerged in the Americas, and since then, little progress has been made to control outbreaks caused by this virus. After its first detection in New York in 1999, West Nile virus quickly spread across the continent, causing an epidemic of human disease and massive bird die-offs. Now the virus has become endemic to the United States, where an estimated 7 million human infections have occurred, making it the leading mosquito-borne virus infection and the most common cause of viral encephalitis in the country. To bring new attention to one of the most important mosquito-borne viruses in the Americas, we provide an interactive review using Nextstrain: a visualization tool for real-time tracking of pathogen evolution (nextstrain.org/WNV/NA). Nextstrain utilizes a growing database of more than 2,000 West Nile virus genomes and harnesses the power of phylogenetics for students, educators, public health workers, and researchers to visualize key aspects of virus spread and evolution. Using Nextstrain, we use virus genomics to investigate the emergence of West Nile virus in the U S, followed by its rapid spread, evolution in a new environment, establishment of endemic transmission, and subsequent international spread. For each figure, we include a link to Nextstrain to allow the readers to directly interact with and explore the underlying data in new ways. We also provide a brief online narrative that parallels this review to further explain the data and highlight key epidemiological and evolutionary features (nextstrain.org/narratives/twenty-years-of-WNV). Mirroring the dynamic nature of outbreaks, the Nextstrain links provided within this paper are constantly updated as new West Nile virus genomes are shared publicly, helping to stay current with the research. Overall, our review showcases how genomics can track West Nile virus spread and evolution, as well as potentially uncover novel targeted control measures to help alleviate its public health burden.


Clinicopathologic characteristics of 143 patients with synchronous bilateral invasive breast carcinomas treated in a single institution.
Synchronous bilateral invasive breast carcinoma (SBIBC) ranged in incidence from 0.3% to as high as 12%. Between April 1997 and February 2003, 143 consecutive patients with SBIBC were treated at the European Institute of Oncology (Milan, Italy). Their information was collected prospectively in a database. The bilateral tumors were divded into left and right tumors. Tumor size, histology, grade, lymph node status, estrogen (ER) and progesterone receptor (PgR) status, HER-2 expression, peritumoral vascular invasion (PVI), Ki-67 expression, extensive in situ component (EIC), and multifocality between the two groups were analyzed. During the same time period, 6218 patients with unilateral invasive breast carcinoma (UIBC) were analyzed in the same manner for comparison with the patients with SBIBC. There were no significant differences between left and right tumors, and the observed histopathologic agreement within the same patient was significantly superior than statistically expected for all characteristics except size, lymph node status, and multifocality. When compared with patients with UIBC, patients with SBIBC were more likely to present with smaller tumors and showed a higher frequency of invasive lobular carcinoma, lower histologic grade, higher rate of ER and PgR positivity, and lower PVI and Ki-67 expression. The high concordance of histopathologic characteristics between SBIBC within the same patient could reflect a particular hormonal environment that influenced either the initiation and development of these lesions simultaneously and independently from the single or multi-clonal origin, either a less aggressive biological behavior compared with UIBC. In particular, the strong agreement of the observed EIC in SBIBC within the same patient seemed to definitively exclude the metastatic origin of these tumors.


Salt-regulated mannitol metabolism in algae.
Mannitol, one of the most widely occurring sugar alcohol compounds, is found in bacteria, fungi, algae, and plants. In these organisms the compound acts as a compatible solute and has multiple functions, including osmoregulation, storage, and regeneration of reducing power, and scavenging of active oxygen species. Because of the diverse functions of mannitol, introducing the ability to accumulate it has been a hallmark of attempts to generate highly salt-tolerant transgenic plants. However, transgenic plants have not yet improved significantly in their salt tolerance. Recently, we purified and characterized 2 enzymes that biosynthesize mannitol, mannitol-1-phosphate dehydrogenase (M1PDH) and mannitol-1-phosphate-specific phosphatase, from the marine red alga Caloglossa continua, which grows in estuarine areas where tide levels fluctuate frequently. The activation of Caloglossa M1PDH is unique in that it is regulated by salt concentration at enzyme level. In this review we focus on the metabolism of mannitol, mainly in marine photosynthetic organisms, and suggest how this might be applied to producing salt-tolerant transgenic plants.


Mistranslation can enhance fitness through purging of deleterious mutations.
Phenotypic mutations are amino acid changes caused by mistranslation. How phenotypic mutations affect the adaptive evolution of new protein functions is unknown. Here we evolve the antibiotic resistance protein TEM-1 towards resistance on the antibiotic cefotaxime in an Escherichia coli strain with a high mistranslation rate. TEM-1 populations evolved in such strains endow host cells with a general growth advantage, not only on cefotaxime but also on several other antibiotics that ancestral TEM-1 had been unable to deactivate. High-throughput sequencing of TEM-1 populations shows that this advantage is associated with a lower incidence of weakly deleterious genotypic mutations. Our observations show that mistranslation is not just a source of noise that delays adaptive evolution. It could even facilitate adaptive evolution by exacerbating the effects of deleterious mutations and leading to their more efficient purging. The ubiquity of mistranslation and its effects render mistranslation an important factor in adaptive protein evolution.


Adherence to clinical practice guidelines and outcomes in diabetic patients.
To examine the level of adherence to clinical practice guidelines and its relationship to outcomes in patients with diabetes. Retrospective cohort study. A tertiary teaching hospital in Korea. Patients aged ≥18 years with diabetes (n = 4994) who visited the study hospital once or more during 2004. The outcomes were mortality from the database of the Statistics Korea and end-stage renal disease (ESRD) incidence from ESRD registry in the Korean Society of Nephrology until December 2009. Testing rates for blood pressure, eye examination, HbA1c, renal function and lipid profiles were 93.9, 32.8, 84.9, 33.5 and 45.9%, respectively. The percentage of patients achieving each treatment goal was 27.8% for blood pressure, 44.2% for HbA1c and 49.4% for low-density lipoprotein (LDL) cholesterol. There were 11.7% patients with composite outcome (death and/or ESRD). Male gender, level of HbA1c (<7%), presence of HbA1c data, checking eye examination, presence of data on urine albumin-to-creatinine ratio (UACR) and having anti-platelet medication were associated with better outcome. The adherence to recommendations was unsatisfactory, especially in checking eye examination, testing UACR and LDL cholesterol, and achieving a target goal for each parameter. Guideline adherence was positively related to better prognosis. Active strategies to apply the guidelines to clinical practice should be developed to improve patient outcomes.


Development of spray deposition/MALDI-TOFMS and its application to the rapid screening of hydrolysis products derived from nitrogen mustards.
A novel method for preparing samples for use in MALDI-TOFMS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) is described. Seven hydrolysis products derived from nitrogen mustards and CHCA (alpha-cyano-4-hydroxycinnamic acid) were selected as model compounds and the matrix, respectively. A capillary atomizer was used for evaporative and spray deposition of the sample/matrix solution, leading to the formation of a freestanding film that coated and accumulated on the MALDI substrate (i.e., sample plate). Compared to the traditional method for MALDI, which involves the production of dried droplets, the surface roughness was reduced, resulting in the accumulation of the sample-doped matrix on the sample plate. This resulted in an increase in the limit of detection of 1 - 2 orders of magnitude. In order to compare the structures of the sample-doped matrices obtained by the traditional dried droplet method versus the spray deposition method (developed in this study), the matrices were examined by SEM (scanning electron microscopy). The design of the capillary atomizer and details of the experimental conditions are reported. The application of this method to the above seven degradation products was successful, suggesting that it has great potential for use as a routine monitoring tool.


Thermal Contrast Amplification Reader Yielding 8-Fold Analytical Improvement for Disease Detection with Lateral Flow Assays.
There is an increasing need for highly sensitive and quantitative diagnostics at the point-of-care. The lateral flow immunoassay (LFA) is one of the most widely used point-of-care diagnostic tests; however, LFAs generally suffer from low sensitivity and lack of quantification. To overcome these limitations, thermal contrast amplification (TCA) is a new method that is based on the laser excitation of gold nanoparticles (GNPs), the most commonly used visual signature, to evoke a thermal signature. To facilitate the clinical translation of the TCA technology, we present the development of a TCA reader, a platform technology that significantly improves the limit of detection and provides quantification of disease antigens in LFAs. This TCA reader provides enhanced sensitivity over visual detection by the human eye or by a colorimetric reader (e.g., BD Veritor System Reader). More specifically, the TCA reader demonstrated up to an 8-fold enhanced analytical sensitivity and quantification among LFAs for influenza, malaria, and Clostridium difficile. Systematic characterization of the laser, infrared camera, and other components of the reader and their integration into a working reader instrument are described. The development of the TCA reader enables simple, highly sensitive quantification of LFAs at the point-of-care.


Current and novel therapeutic approaches in myelodysplastic syndromes.
Myelodysplastic syndromes (MDS) are a heterogeneous group of hematologic neoplasms with an annual incidence of 4.1 cases per 100,000 Americans. Patients with MDS suffer from chronic cytopenias that may lead to recurrent transfusions, infections, and increased risk for bleeding. They are also at risk for progression to acute myeloid leukemia. Allogeneic hematopoietic cell transplantation is the only potentially curative treatment for MDS, although 3 drugs have been approved by the US Food and Drug Administration for its treatment: lenalidomide, 5-azacitidine, and decitabine. These therapies can be effective in the relief of cytopenias, achievement of cytogenetic remissions, and reduction in bone marrow blasts. 5-azacitidine has also been shown to improve overall survival. However, there remain many unmet needs in the treatment of MDS. Breakthroughs in our understanding of the complex pathogenesis of MDS through epigenetic, genetic, immunologic, and other biological mechanisms have allowed us to develop new therapeutic strategies that can lead to improvements in outcomes in MDS. In this review, we aim to provide an overview of the evolution in classifcation and risk stratifcation in MDS and to illustrate how we can use this to guide us in tailoring therapeutic choices in this disease. Responses and outcomes related to com monly used MDS therapies will be discussed together with novel therapies that have evolved with the improved understanding of MDS pathophysiology.


Chelation by histidine inhibits the vacuolar sequestration of nickel in roots of the hyperaccumulator Thlaspi caerulescens.
* The mechanisms of enhanced root to shoot metal transport in heavy metal hyperaccumulators are incompletely understood. Here, we compared the distribution of nickel (Ni) over root segments and tissues in the hyperaccumulator Thlaspi caerulescens and the nonhyperaccumulator Thlaspi arvense, and investigated the role of free histidine in Ni xylem loading and Ni transport across the tonoplast. * Nickel accumulation in mature cortical root cells was apparent in T. arvense and in a high-Ni-accumulating T. caerulescens accession, but not in a low-accumulating T. caerulescens accession. * Compared with T. arvense, the concentration of free histidine in T. caerulescens was 10-fold enhanced in roots, but was only slightly higher in leaves, regardless of Ni exposure. Nickel uptake in MgATP-energized root- and shoot-derived tonoplast vesicles was almost completely blocked in T. caerulescens when Ni was supplied as a 1 : 1 Ni-histidine complex, but was uninhibited in T. arvense. Exogenous histidine supply enhanced Ni xylem loading in T. caerulescens but not in T. arvense. * The high rate of root to shoot translocation of Ni in T. caerulescens compared with T. arvense seems to depend on the combination of two distinct characters, that is, a greatly enhanced root histidine concentration and a strongly decreased ability to accumulate histidine-bound Ni in root cell vacuoles.


Impact of p53 loss on reversal and recurrence of conditional Wnt-induced tumorigenesis.
Aberrant activation of Wnt signaling is oncogenic and has been implicated in a variety of human cancers. We have developed a doxycycline-inducible Wnt1 transgenic mouse model to determine the dependence of established mammary adenocarcinomas on continued Wnt signaling. Using this model we show that targeted down-regulation of the Wnt pathway results in the rapid disappearance of essentially all Wnt-initiated invasive primary tumors as well as pulmonary metastases. Tumor regression does not require p53 and occurs even in highly aneuploid tumors. However, despite the dependence of primary mammary tumors and metastases on continued Wnt signaling and the dispensability of p53 for tumor regression, we find that a substantial fraction of tumors progress to a Wnt-independent state and that p53 suppresses this process. Specifically, loss of one p53 allele dramatically facilitates the progression of mammary tumors to a Wnt1-independent state both by impairing the regression of primary tumors following doxycycline withdrawal and by promoting the recurrence of fully regressed tumors in the absence of doxycycline. Thus, although p53 itself is dispensable for tumor regression, it nevertheless plays a critical role in the suppression of tumor recurrence. Our findings demonstrate that although even advanced stages of epithelial malignancy remain dependent upon continued Wnt signaling for maintenance and growth, loss of p53 facilitates tumor escape and the acquisition of oncogene independence.


Task-Based Effectiveness of Basic Visualizations.
Visualizations of tabular data are widely used; understanding their effectiveness in different task and data contexts is fundamental to scaling their impact. However, little is known about how basic tabular data visualizations perform across varying data analysis tasks. In this paper, we report results from a crowdsourced experiment to evaluate the effectiveness of five small scale (5-34 data points) two-dimensional visualization types-Table, Line Chart, Bar Chart, Scatterplot, and Pie Chart-across ten common data analysis tasks using two datasets. We find the effectiveness of these visualization types significantly varies across task, suggesting that visualization design would benefit from considering context-dependent effectiveness. Based on our findings, we derive recommendations on which visualizations to choose based on different tasks. We finally train a decision tree on the data we collected to drive a recommender, showcasing how to effectively engineer experimental user data into practical visualization systems.


Weeding out the information: an ethnographic approach to exploring how young people make sense of the evidence on cannabis.
Contradictory evidence on cannabis adds to the climate of confusion regarding the health harms related to use. This is particularly true for young people as they encounter and make sense of opposing information on cannabis. Knowledge translation (KT) is in part focused on ensuring that knowledge users have access to and understand best evidence; yet, little attention has focused on the processes youth use to weigh scientific evidence. There is growing interest in how KT efforts can involve knowledge users in shaping the delivery of youth-focused public health messages. To date, the youth voice has been largely absent from the creation of public health messages on cannabis. This ethnographic study describes a knowledge translation project that focused on engaging young people in a review of evidence on cannabis that concluded with the creation of public health messages generated by youth participants. We facilitated two groups with a total of 18 youth participants. Data included transcribed segments of weekly sessions, researcher field notes, participant research logs, and transcribed follow-up interviews. Qualitative, thematic analysis was conducted. Group dynamics were influential in terms of how participants made sense of the evidence. The processes by which participants came to understand the current evidence on cannabis are described, followed by the manner in which they engaged with the literature for the purpose of creating an individual public health message to share with the group. At project end, youth created collaborative public health messages based on their understanding of the evidence illustrating their capacity to "weed out" the information. The content of these messages reflect a youth-informed harm reduction approach to cannabis use. This study demonstrates the feasibility of involving young people in knowledge translation initiatives that target peers. Youth participants demonstrated that they were capable of reading scientific literature and had the capacity to engage in the creation of evidence-informed public health messages on cannabis that resonate with young people. Rather than simply being the target of KT messages, they embraced the opportunity to engage in dialogue focused on cannabis.


The future for fisheries.
Formal analyses of long-term global marine fisheries prospects have yet to be performed, because fisheries research focuses on local, species-specific management issues. Extrapolation of present trends implies expansion of bottom fisheries into deeper waters, serious impact on biodiversity, and declining global catches, the last possibly aggravated by fuel cost increases. Examination of four scenarios, covering various societal development choices, suggests that the negative trends now besetting fisheries can be turned around, and their supporting ecosystems rebuilt, at least partly.


Total testosterone and DHEAS levels as predictors of androgen-secreting neoplasms: a populational study.
Androgen excess affects between 2% and 10% of women. While the majority of these patients suffer from polycystic ovary syndrome, a few present with an androgen-secreting neoplasm. An elevated circulating total testosterone level and dehydroepiandiosterone sulfate (DHEAS) level have been proposed as screening methods for detecting ovarian and adrenal androgen-secreting neoplasms, respectively. To determine the predictive value of these tests for androgen-secreting tumors in a population of consecutive hyperandrogenic patients, we studied 478 consecutive untreated hyperandrogenic patients presenting over a ten-year period (1987-97). All had at least two of the following features: (1) oligomenorrhea (i.e. cycles > 35 days or < 8 cycles/year), (2) hyperandrogenemia (i.e. a total or free testosterone, or DHEAS > 95th percentile of controls), or (3) hirsutism (i.e. a modified Ferriman-Gallwey score > or = 6). None of these patients had a prior diagnosis of an androgen-secreting neoplasm. Basal levels of testosterone and DHEAS were determined in all patients, with transvaginal sonography and an adrenal computed tomography scan in select individuals. Of the 478 patients included, 65% had hirsutism and oligomenorrhea; 20% had hyperandrogenic oligomenorrhea; and 15% had hirsutism and hyperandrogenemia, without overt oligomenorrhea. Overall, 11 (2.3%) patients had a total testosterone > 8.7 nmol/l (250 ng/dl), of which one actually had an androgen-secreting neoplasm (i.e. true-positive). This postmenopausal patient presented with rapidly progressive virilization, and demonstrated an ovarian hilar cell tumor at surgery. The calculated sensitivity of an elevated testosterone level (> 8.67 nmol/l) for a neoplasm was 100% (1/1), the specificity was 98% (467/477), and the negative predictive value was 100% (467/467), but the positive predictive value was only 9% (1/11). Ten subjects had DHEAS levels > 16.3 mumol/l (6000 ng/ml), and none was diagnosed with an adrenocortical tumor. Although the sensitivity and positive predictive value of a high DHEAS for a neoplasm could not be calculated due to the absence of a test case, the specificity was 98% (468/478) and the negative predictive value was 100% (468/468). These data suggest that the measurement of testosterone and DHEAS is not a cost-effective method of screening for these tumors, due to the low frequency of the disorder and the fact that clinical evaluation alone is often sufficient screening.


Perceiving emotional expressions in others: Activation likelihood estimation meta-analyses of explicit evaluation, passive perception and incidental perception of emotions.
We conducted a series of activation likelihood estimation (ALE) meta-analyses to determine the commonalities and distinctions between separate levels of emotion perception, namely incidental perception, passive perception, and explicit evaluation of emotional expressions. Pooling together more than 180 neuroimaging experiments using facial, vocal or body expressions, our results are threefold. First, explicitly evaluating the emotions of others recruits brain regions associated with the sensory processing of expressions, such as the inferior occipital gyrus, middle fusiform gyrus and the superior temporal gyrus, and brain regions involved in low-level and high-level mindreading, namely the posterior superior temporal sulcus, the inferior frontal cortex and dorsomedial frontal cortex. Second, we show that only the sensory regions were also consistently active during the passive perception of emotional expressions. Third, we show that the brain regions involved in mindreading were active during the explicit evaluation of both facial and vocal expressions. We discuss these results in light of the existing literature and conclude by proposing a cognitive model for perceiving and evaluating the emotions of others.


Association Between the Use of Antidepressants and the Risk of Type 2 Diabetes: A Large, Population-Based Cohort Study in Japan.
This study aimed to reveal the associations between the risk of new-onset type 2 diabetes and the duration of antidepressant use and the antidepressant dose, and between antidepressant use after diabetes onset and clinical outcomes. In this large-scale retrospective cohort study in Japan, new users of antidepressants (exposure group) and nonusers (nonexposure group), aged 20-79 years, were included between 1 April 2006 and 31 May 2015. Patients with a history of diabetes or receipt of antidiabetes treatment were excluded. Covariates were adjusted by using propensity score matching; the associations were analyzed between risk of new-onset type 2 diabetes and the duration of antidepressant use/dose of antidepressant in the exposure and nonexposure groups by using Cox proportional hazards models. Changes in glycated hemoglobin (HbA1c) level were examined in groups with continuous use, discontinuation, or a reduction in the dose of antidepressants. Of 90,530 subjects, 45,265 were in both the exposure and the nonexposure group after propensity score matching; 5,225 patients (5.8%) developed diabetes. Antidepressant use was associated with the risk of diabetes onset in a time- and dose-dependent manner. The adjusted hazard ratio was 1.27 (95% CI 1.16-1.39) for short-term low-dose and 3.95 (95% CI 3.31-4.72) for long-term high-dose antidepressant use. HbA1c levels were lower in patients who discontinued or reduced the dose of antidepressants (F[2,49] = 8.17; P < 0.001). Long-term antidepressant use increased the risk of type 2 diabetes onset in a time- and dose-dependent manner. Glucose tolerance improved when antidepressants were discontinued or the dose was reduced after diabetes onset.


Aeroallergen-induced eosinophilic inflammation, lung damage, and airways hyperreactivity in mice can occur independently of IL-4 and allergen-specific immunoglobulins.
In this investigation we have used a mouse model containing certain phenotypic characteristics consistent with asthma and IL-4- and CD40-deficient mice to establish the role of this cytokine and allergen-specific immunoglobulins in the initiation of airways hyperreactivity and morphological changes to the airways in responses to aeroallergen challenge. Sensitization and aerosol challenge of mice with ovalbumin resulted in a severe airways inflammatory response which directly correlated with the induction of extensive airways damage and airways hyperreactivity to beta-methacholine. Inflammatory infiltrates were primarily characterized by the presence of CD4+ T cells and eosinophils. In IL-4-deficient mice, the recruitment of airways eosinophils was impaired, but not abolished in response to aeroallergen. Moreover, the characteristic airways damage and hyperreactivity normally resulting from allergen inhalation were not attenuated. Induction of these structural and functional changes to the airways occurred in the absence of ovalbumin-specific IgE and IgG1, but IgG2a and IgG3 were detected in the sera of IL-4-deficient mice. CD4+ T cells isolated from both wild-type and IL-4-deficient mice given ovalbumin produced significant levels of IL-5 after in vitro stimulation. Treatment of IL-4-deficient mice with anti-IL-5 mAb before aeroallergen challenge abolished blood and airways eosinophilia, lung damage, and airways hyperreactivity. These results indicate that IL-4 is not essential for the development of IL-5-producing CD4+ T cells or for the induction of eosinophilic inflammation and airways damage and hyperreactivity. In response to sensitization and aerosol challenge, CD40-deficient mice did not produce ovalbumin-specific IgE, IgG isotypes, or IgA, and airways inflammation and hyperreactivity were not attenuated. Our results suggest that allergic airways disease can occur via pathways which operate independently of IL-4 and allergen-specific immunoglobulins. Activation of these pathways is intimately associated with IL-5 and eosinophilic inflammation. Such pathways may play a substantive role in the etiology of asthma.


Coherent spin control of s-, p-, d- and f-electrons in a silicon quantum dot.
Once the periodic properties of elements were unveiled, chemical behaviour could be understood in terms of the valence of atoms. Ideally, this rationale would extend to quantum dots, and quantum computation could be performed by merely controlling the outer-shell electrons of dot-based qubits. Imperfections in semiconductor materials disrupt this analogy, so real devices seldom display a systematic many-electron arrangement. We demonstrate here an electrostatically confined quantum dot that reveals a well defined shell structure. We observe four shells (31 electrons) with multiplicities given by spin and valley degrees of freedom. Various fillings containing a single valence electron-namely 1, 5, 13 and 25 electrons-are found to be potential qubits. An integrated micromagnet allows us to perform electrically-driven spin resonance (EDSR), leading to faster Rabi rotations and higher fidelity single qubit gates at higher shell states. We investigate the impact of orbital excitations on single qubits as a function of the dot deformation and exploit it for faster qubit control.


Versican V0 and V1 direct the growth of peripheral axons in the developing chick hindlimb.
Peanut agglutinin-binding disaccharides and chondroitin sulfate mark transient mesenchymal barriers to advancing motor and sensory axons innervating the hindlimbs during chick development. Here we show that the vast majority of these carbohydrates are at the critical stage and location attached to the versican splice variants V0 and V1. We reveal that the isolated isoforms of this extracellular matrix proteoglycan suppress axon extension at low concentrations and induce growth cone collapse and rapid retraction at higher levels. Moreover, we demonstrate that versican V0 and/or V1, recombinantly expressed in collagen-I gels or ectopically deposited in the hindlimbs of chicken embryos in ovo, cause untimely defasciculation and axon stalling. Consequently, severe disturbances of nerve patterning are observed in the versican-treated embryos. Our experiments emphasize the inhibitory capacity of versicans V0 and V1 in axonal growth and evidence for their function as basic guidance cues during development of the peripheral nervous system.


Auditory event-related potentials (P300) in the identification of psychogenic nonepileptic seizures.
The feasibility and conceivable value of postictal event-related potential (ERP) recordings were studied in patients with nonepileptic seizures (NES) admitted for long-term video/EEG monitoring. Ten patients with NES underwent preictal (on hospital admission) and postictal (< or =6 hours after seizure) ERP recordings of an auditory oddball paradigm. Additionally, 10 temporal lobe epilepsy (TLE) patients with partial seizures and secondary generalization underwent preictal, postictal (< 6 hours after seizures), and interictal (7-48 hours after seizure) ERP recordings. We recently reported that ERPs recorded in TLE patients with partial epilepsy undergo a temporary change postictally, while returning to their preictal state during interictal recordings. In the current study intraclass correlations, transformed into z scores, are used to determine test-retest validity of repeated ERP recordings. An independent sample t test with z scores for the comparison of preictal and postictal recordings showed that ERP activation differed between NES and TLE patients (P=0.009). More specifically, ERP recordings in the preictal and postictal states were similar in NES patients, but dissimilar in TLE patients. On the other hand, this dissimilarity in ERPs disappeared when comparing z scores for the preictal and postictal recordings in NES patients with z scores for the preictal and interictal recordings in TLE patients. This further supports the notion that identical waveforms during preictal and postictal recordings in NES patients reflect nonepileptic seizure activity. The current findings suggest that postictal ERP recordings are useful in the diagnosis of NES and differentiate TLE from NES.


The Challenges and Achievements in the Implementation of the Natural Childbirth Instruction Program: A Qualitative Study.
The natural childbirth instruction program, which aims to reduce the cesarean section (C-section) rates in the country and pay attention to demographic policies, has achieved significant accomplishments in the short time span since it was implemented throughout the country. In the present study, the advantages and challenges of the implementation of this program have been analyzed. This qualitative study carried out with the participation of 32 knowledgeable individuals who were selected using purposeful sampling and snowball sampling methods among the personnel of Kerman University of Medical Sciences, and it's affiliated educational (university) hospital. The data were collected through semi-structured interviews based on the research objectives, review of texts, and experts' experiences. Data analysis was performed using content analysis method in MaxQDA software. Data analysis provided the 5 main categories of implementation instructions' strengths, implementation instructions' defects, implementation instructions' achievements, implementation instructions' challenges and threats, and suggestions. Accountability in the system that provides health care services for pregnant mothers in public hospitals has been created through the implementation of the natural childbirth promotion package. If managerial barriers and executive and legal inefficiencies are followed up and suitable measures are taken for solving the intra-system conflicts, we can hope that the package, which has been one of the most serious efforts made by the Ministry of Health over the past decades to reduce cesarean delivery, will achieve significant accomplishments.


Increase of plasma insulin by racecadotril, an inhibitor of enkephalinase, in Wistar rats.
Racecadotril is known as an inhibitor of enkephalinase. Increase of plasma insulin by racecadotril has been observed in rats while the mechanism of the action remains obscure. In the present study, intravenous injection of male Wistar rats with racecadotril significantly decreased blood glucose levels. However, this effect of racecadotril was not modified by naloxone at the dose sufficient to block opioid receptors. Thus, the blood glucose-lowering action of racecadotril might be through an endogenous opioid independent mechanism. Otherwise, we found that C-peptide content was also raised by racecadotril in parallel with the increase of insulin in Wistar rats. Thus, the blood glucose-lowering action of racecadotril was related to insulin secretion, but not through the inhibition of plasma insulin degradation. In addition, racecadotril showed no direct effect on insulin secretion in isolated islets or cultured HIT-T15 beta cells. The increase of plasma insulin and blood glucose-lowering action induced by racecadotril were reduced by pretreatment with atropine and enhanced by physotigmine. Direct inhibition of cholinesterase was not observed in brain homogenates treated with racecadotril. Moreover, actions of racecadotril were significantly reduced in rats receiving hemicholinium-3 at a sufficient dose to decrease endogenous acetylcholine. Activation of cholinergic tone is possibly involved in the blood glucose-lowering effect of racecadotril. Our results suggested that racecadotril increased insulin secretion to lower blood glucose mainly via regulation of parasympathetic tone in Wistar rats.


STITCH: interaction networks of chemicals and proteins.
The knowledge about interactions between proteins and small molecules is essential for the understanding of molecular and cellular functions. However, information on such interactions is widely dispersed across numerous databases and the literature. To facilitate access to this data, STITCH ('search tool for interactions of chemicals') integrates information about interactions from metabolic pathways, crystal structures, binding experiments and drug-target relationships. Inferred information from phenotypic effects, text mining and chemical structure similarity is used to predict relations between chemicals. STITCH further allows exploring the network of chemical relations, also in the context of associated binding proteins. Each proposed interaction can be traced back to the original data sources. Our database contains interaction information for over 68,000 different chemicals, including 2200 drugs, and connects them to 1.5 million genes across 373 genomes and their interactions contained in the STRING database. STITCH is available at http://stitch.embl.de/.


Impact of genetic notification on smoking cessation: systematic review and pooled-analysis.
This study aimed to evaluate the impact of genetic notification of smoking-related disease risk on smoking cessation in the general population. Secondary objectives were to assess the impact of genetic notification on intention-to-quit smoking and on emotional outcomes as well as the understanding and the recall of this notification. A systematic review of articles from inception to August 2011 without language restriction was realized using PubMed, Embase, Scopus, Web of Science, PsycINFO and Toxnet. Other publications were identified using hand search. The pooled-analysis included only randomized trials. Comparison groups were (i) high and low genetic risk versus control, and (ii) high versus low genetic risk. For the pooled-analysis random effect models were applied and sensitivity analyses were conducted. Eight papers from seven different studies met the inclusion criteria of the review. High genetic risk notification was associated with short-term increased depression and anxiety. Four randomized studies were included in the pooled-analysis, which revealed a significant impact of genetic notification on smoking cessation in comparison to controls (clinical risk notification or no intervention) in short term follow-up less than 6 months (RR = 1.55, 95% CI 1.09-2.21). In short term follow-up, genetic notification increased smoking cessation in comparison to control interventions. However, there is no evidence of long term effect (up to 12 month) on smoking cessation. Further research is needed to assess more in depth how genetic notification of smoking-related disease could contribute to smoking cessation.


DNA Copy Number Aberrations, and Human Papillomavirus Status in Penile Carcinoma. Clinico-Pathological Correlations and Potential Driver Genes.
Penile squamous cell carcinoma is a rare disease, in which somatic genetic aberrations have yet to be characterized. We hypothesized that gene copy aberrations might correlate with human papillomavirus status and clinico-pathological features. We sought to determine the spectrum of gene copy number aberrations in a large series of PSCCs and to define their correlations with human papillomavirus, histopathological subtype, and tumor grade, stage and lymph node status. Seventy formalin-fixed, paraffin embedded penile squamous cell carcinomas were centrally reviewed by expert uropathologists. DNA was extracted from micro-dissected samples, subjected to PCR-based human papillomavirus assessment and genotyping (INNO-LiPA human papillomavirus Genotyping Extra Assay) and microarray-based comparative genomic hybridization using a 32K Bacterial Artificial Chromosome array platform. Sixty-four samples yielded interpretable results. Recurrent gains were observed in chromosomes 1p13.3-q44 (88%), 3p12.3-q29 (86%), 5p15.33-p11 (67%) and 8p12-q24.3 (84%). Amplifications of 5p15.33-p11 and 11p14.1-p12 were found in seven (11%) and four (6%) cases, respectively. Losses were observed in chromosomes 2q33-q37.3 (86%), 3p26.3-q11.1 (83%) and 11q12.2-q25 (81%). Although many losses and gains were similar throughout the cohort, there were small significant differences observed at specific loci, between human papillomavirus positive and negative tumors, between tumor types, and tumor grade and nodal status. These results demonstrate that despite the diversity of genetic aberrations in penile squamous cell carcinomas, there are significant correlations between the clinico-pathological data and the genetic changes that may play a role in disease natural history and progression and highlight potential driver genes, which may feature in molecular pathways for existing therapeutic agents.


Lamb wave propagation in monocrystalline silicon wafers.
Monocrystalline silicon wafers are widely used in the photovoltaic industry for solar panels with high conversion efficiency. Guided ultrasonic waves offer the potential to efficiently detect micro-cracks in the thin wafers. Previous studies of ultrasonic wave propagation in silicon focused on effects of material anisotropy on bulk ultrasonic waves, but the dependence of the wave propagation characteristics on the material anisotropy is not well understood for Lamb waves. The phase slowness and beam skewing of the two fundamental Lamb wave modes A0 and S0 were investigated. Experimental measurements using contact wedge transducer excitation and laser measurement were conducted. Good agreement was found between the theoretically calculated angular dependency of the phase slowness and measurements for different propagation directions relative to the crystal orientation. Significant wave skew and beam widening was observed experimentally due to the anisotropy, especially for the S0 mode. Explicit finite element simulations were conducted to visualize and quantify the guided wave beam skew. Good agreement was found for the A0 mode, but a systematic discrepancy was observed for the S0 mode. These effects need to be considered for the non-destructive testing of wafers using guided waves.


Playground drinking fountains in 17 local government areas: survey methods and results.
Appropriate public access to water is an increasing concern, and to further explore field observation methods for assessing such access, we aimed to survey drinking fountains in playgrounds across many local government areas. We systematically collected data (including photographs) of drinking fountains in randomly selected public playgrounds in 17 local government areas (TLAs) in New Zealand. The time for playground surveys was always less than 15 minutes. We found only one of the 17 TLAs had working drinking fountains in all the playgrounds sampled, and 11 working fountains in all 54 playgrounds (20%). Three had metal discolouration within 1cm of the nozzle. The systematic observation method was relatively quick, making it suitable for local officials and health promoters.


Towards standardized measurement of adverse events in spine surgery: conceptual model and pilot evaluation.
Independent of efficacy, information on safety of surgical procedures is essential for informed choices. We seek to develop standardized methodology for describing the safety of spinal operations and apply these methods to study lumbar surgery. We present a conceptual model for evaluating the safety of spine surgery and describe development of tools to measure principal components of this model: (1) specifying outcome by explicit criteria for adverse event definition, mode of ascertainment, cause, severity, or preventability, and (2) quantitatively measuring predictors such as patient factors, comorbidity, severity of degenerative spine disease, and invasiveness of spine surgery. We created operational definitions for 176 adverse occurrences and established multiple mechanisms for reporting them. We developed new methods to quantify the severity of adverse occurrences, degeneration of lumbar spine, and invasiveness of spinal procedures. Using kappa statistics and intra-class correlation coefficients, we assessed agreement for the following: four reviewers independently coding etiology, preventability, and severity for 141 adverse occurrences, two observers coding lumbar spine degenerative changes in 10 selected cases, and two researchers coding invasiveness of surgery for 50 initial cases. During the first six months of prospective surveillance, rigorous daily medical record reviews identified 92.6% of the adverse occurrences we recorded, and voluntary reports by providers identified 38.5% (surgeons reported 18.3%, inpatient rounding team reported 23.1%, and conferences discussed 6.1%). Trained observers had fair agreement in classifying etiology of 141 adverse occurrences into 18 categories (kappa = 0.35), but agreement was substantial (kappa > or = 0.61) for 4 specific categories: technical error, failure in communication, systems failure, and no error. Preventability assessment had moderate agreement (mean weighted kappa = 0.44). Adverse occurrence severity rating had fair agreement (mean weighted kappa = 0.33) when using a scale based on the JCAHO Sentinel Event Policy, but agreement was substantial for severity ratings on a new 11-point numerical severity scale (ICC = 0.74). There was excellent inter-rater agreement for a lumbar degenerative disease severity score (ICC = 0.98) and an index of surgery invasiveness (ICC = 0.99). Composite measures of disease severity and surgery invasiveness may allow development of risk-adjusted predictive models for adverse events in spine surgery. Standard measures of adverse events and risk adjustment may also facilitate post-marketing surveillance of spinal devices, effectiveness research, and quality improvement.


Association of interleukin-10 with hepatitis B virus (HBV) mediated disease progression in Indian population.
Interleukin (IL)-10, an anti-inflammatory Th2 cytokine, is one of the key coordinators of the inflammatory responses involved. The present study was designed to evaluate the impact of IL-10 (-819/-592) genotypes, haplotypes, mRNA and the protein levels with risk for hepatitis B virus (HBV) related hepatocellular carcinoma (HCC) development in India. A total of 390 subjects (145 controls, 62 inactive HBV-carriers, 64 chronic-active HBV patients, 60 HBV related cirrhotics and 59 HBV- HCC subjects) were enrolled in the study. Allele specific (AS)-PCR, ELISA and RT-PCR methods were used for assessing polymorphism, spontaneous blood levels and the mRNA expression, respectively of IL-10. The study revealed that the CC/TA genotype acted as a risk factor for cirrhosis (OR a =2.02; P<0.05) and the subsequent HCC development (OR a =2.20; P<0.05), with controls as reference. However, no significant association was found between the two haplotypes (CC and TA) observed and HCC risk. Moreover, the IL-10 protein and mRNA levels in peripheral blood mono nuclear cells (PBMCs) showed a significant elevation as the disease progressed to cirrhosis. But, no variation was observed in the IL-10 levels in subjects with different IL-10 genotypes. These preliminary results suggest a strong association of IL10 (-819/-592) with the HBV infection mediated disease progression, from inactive carrier state to malignancy, in Indian population.


Quantitative phase-contrast imaging through a scattering media.
We report a new technique for the recovery of quantitative phase and amplitude information of an object hidden behind a scattering medium. Two-point intensity correlation measurement together with digital holography principles are utilized for this purpose. The hologram information of the object and a reference beam is scrambled by the presence of a scattering medium in its path. A direct digital holographic recording of this scattered light does not lead to the reconstruction of actual object information. We propose the idea of recovering this hologram information from the spatially fluctuating field of a laser speckle pattern using the intensity correlation, and subsequently apply digital reconstruction of the hologram for recovery of quantitative phase and amplitude information of objects hidden by a random diffuser.


CTGF Contributes to the Development of Posterior Capsule Opacification: an in vitro and in vivo study.
Connective tissue growth factor (CTGF) is a crucial factor that plays a major role in the process of posterior capsule opacification (PCO). However, the effects of CTGF on the proliferation and migration of lens epithelial cells (LECs) and on the mechanism of the epithelial mesenchymal transition (EMT) and extracellular matrix (ECM) in human lens epithelial cells (HLECs) as well as the effects of shRNA-mediated CTGF knockdown on the development of PCO in rats remain unclear. In the present study, we found that CTGF promoted EMT, proliferation, migration and the expression of p-ERK1/2 protein in HLECs but exerted little effect on the expression of p-p38 and p-JNK1/2 proteins. MEK inhibitor U0126 effectively restrained the CTGF-induced expression of α-smooth muscle actin (α-SMA), fibronectin (Fn) and type I collagen (COL-1) in HLECs. CTGF knockdown effectively postponed the onset of PCO in the rats and significantly reduced the expression of α-SMA in the capsule. In conclusion, CTGF contributed to the development of PCO presumably by promoting proliferation, migration of LECs, EMT specific protein expression and ECM synthesis in HLECs, which is dependent on ERK signalling. Furthermore, blocking CTGF effectively inhibited PCO in the rats and the EMT specific protein expression in the lens capsule.


Genetic predisposition to breast cancer.
Breast cancer risk factors are closely intertwined with the patient's cultural background, which may contribute to breast cancer aggregations within families. The difficult questions are: (1) does a truly hereditary breast cancer subset exist; (2) which familial aggregations are hereditary; and (3) is the hereditary form distinctive from its sporadic counterpart? These queries will be resolved once biomarkers are identified that show high sensitivity and specificity with genotype. The authors provide a review of this subject and will focus on their recent discovery of increased in vitro hyperdiploidy in cultured skin fibroblasts from patients with or at risk for hereditary breast cancer. The authors discuss findings from their study of family histories in 225 consecutively ascertained patients with verified breast cancer from the Creighton University School of Medicine Oncology Clinic. Findings consistent with an hereditary breast cancer syndrome were identified in 5% of the patients. Given the 112,000 new cases of breast cancer in the United States in 1982, the authors estimate that with a confidence coefficient of 0.95 between 2410 and 8790 of these individuals will manifest hereditary breast cancer. Specific surveillance/management programs should be geared to high-risk members of these families in which cancer yield will be predictable.


In vivo fluorescent optical imaging of cytotoxic T lymphocyte migration using IRDye800CW near-infrared dye.
We describe a method to measure in vivo migration of human T cells by using the near-infrared (NIR) dye IRDye800CW. Labeling of Epstein-Barr virus-specific T cells with IRDye800CW did not affect viability, proliferation, or T cell function. Following tail vein injection into mice bearing subcutaneous tumors, the NIR signal could be measured in vivo at the tumor site. Analysis of tumors revealed T cell infiltration and an increased NIR signal, confirming T cell migration. To test specific migration with IRDye800CW, tumors were modified to express CCL5 to measure site-specific migration. The NIR signal was increased at CCL5-secreting tumors compared with control tumors. Together, these data suggest that IRDye800CW may be used to study the trafficking of T cells in a small animal model and may have potential as a short-term reporter molecule for human immunotherapy studies.


Risk factors for malaria in high incidence areas of Viet Nam: a case-control study.
A key step to advancing the goal of malaria elimination in Viet Nam by 2030 is focusing limited resources for treatment and prevention to groups most at risk for malaria transmission. To better understand risk factors for malaria transmission in central Viet Nam, a survey of 1000 malaria positive cases and 1000 malaria negative controls was conducted. Cases and controls were matched for age and gender and self-presented at commune health stations (CHS) in Binh Phuoc, Dak Nong and Dak Lak Provinces. Diagnoses were confirmed with microscopy, rapid diagnostic test and PCR. Participants were interviewed about 50 potential risk factors for malaria, which included information about occupation, forest visitation, travel, healthcare-seeking behaviour and prior use of anti-malaria interventions. Participants were enrolled by trained government health workers and the samples were analysed in Vietnamese government laboratories. Data were analysed by univariable, block-wise and multivariable logistic regression. Among cases, 61.8% had Plasmodium falciparum, 35.2% Plasmodium vivax and 3% mixed species infections. Median (IQR) age was 27 (21-36) years and 91.2% were male. Twenty-five risk factors were associated with being a case and eleven with being a control. Multivariable analysis found that malaria cases correlated with forest workers, recent forest visitation, longer duration of illness, having a recorded fever, number of malaria infections in the past year, having had prior malaria treatment and having previously visited a clinic. This study demonstrates the benefits of increased statistical power from matched controls in malaria surveillance studies, which allows identification of additional independent risk factors. It also illustrates an example of research partnership between academia and government to collect high quality data relevant to planning malaria elimination activities. Modifiable risk factors and implications of the findings for malaria elimination strategy are presented.


Symbionts as major modulators of insect health: lactic acid bacteria and honeybees.
Lactic acid bacteria (LAB) are well recognized beneficial host-associated members of the microbiota of humans and animals. Yet LAB-associations of invertebrates have been poorly characterized and their functions remain obscure. Here we show that honeybees possess an abundant, diverse and ancient LAB microbiota in their honey crop with beneficial effects for bee health, defending them against microbial threats. Our studies of LAB in all extant honeybee species plus related apid bees reveal one of the largest collections of novel species from the genera Lactobacillus and Bifidobacterium ever discovered within a single insect and suggest a long (>80 mya) history of association. Bee associated microbiotas highlight Lactobacillus kunkeei as the dominant LAB member. Those showing potent antimicrobial properties are acquired by callow honey bee workers from nestmates and maintained within the crop in biofilms, though beekeeping management practices can negatively impact this microbiota. Prophylactic practices that enhance LAB, or supplementary feeding of LAB, may serve in integrated approaches to sustainable pollinator service provision. We anticipate this microbiota will become central to studies on honeybee health, including colony collapse disorder, and act as an exemplar case of insect-microbe symbiosis.


Advanced strategies for treatment of Parkinson's disease: the role of early treatment.
Parkinson's disease (PD) is associated with significant patient disability and costs to the healthcare system. It is questioned whether early treatment may improve outcomes and delay disability. Early treatment relies on early diagnosis, which can be difficult to achieve because the diagnosis of PD is based on motor symptoms, is clinical in nature, and is complicated by potential presentation of nonmotor symptoms prior to motor symptoms. Economic analyses demonstrate that treatments other than levodopa may be cost-effective. The lack of correlation between Unified PD Rating Scale (UPDRS) outcomes and imaging studies of dopamine uptake may reflect the inappropriate selection of study end points, since activities of daily living scores may be more applicable than motor function scores. Levodopa, the standard therapy for motor control of PD and one of the most effective options, is associated with complications (a wearing-off effect) when used long term. Other therapies, including dopamine agonists and monoamine oxidase type-B (MAO-B) inhibitors, may limit the rate of dyskinesia relative to levodopa-based regimens. It appears that early treatment with the MAO-B inhibitor rasagiline (1 mg), as compared with late treatment, delays the onset of worsened UPDRS score, especially the nonmotor activities of daily living subscore.


The effects of psychosocial stress on item, cued-pair and emotional memory.
Physical stress, such as from the cold-pressor test, has been robustly associated with altered memory retrieval, but it is less clear whether the same happens following psychosocial stress. Studies using psychosocial stressors report mixed effects on memory, leading to uncertainty about the common cognitive impact of both forms of stress. The current study uses a series of four carefully designed experiments, each differing by only a single critical factor to determine the effects of psychosocial stress on specific aspects of episodic memory. In three experiments, we induced psychosocial stress after participants encoded words, then assessed retrieval of those words after a prolonged delay. These experiments found no effect of post-encoding stress on recognition of neutral words or cued recall of word-pairs, but a small effect on recollection of semantically related words. There were, however, positive relationships within the stress group between measures of stress (cortisol in experiment 1 and self-reported-anxiety in experiment 3) and recollection of single word stimuli. In the fourth experiment, we found that psychosocial stress immediately before retrieval did not influence word recognition. Recollection, particularly for semantically related stimuli, may therefore be more susceptible to the effects of psychosocial stress, and future studies can assess how this relates to other forms of stress. Overall, our findings suggest that the effects of psychosocial stress on episodic memory may be more subtle than expected, warranting further exploration in larger studies.


Altered myocardial force-frequency relation in human heart failure.
In congestive heart failure (idiopathic dilated cardiomyopathy), exercise is accompanied by a smaller-than-normal decrease in end-diastolic left ventricular volume, depressed peak rates of left ventricular pressure rise and fall, and depressed heart-rate-dependent potentiation of contractility (bowditch treppe). We studied contractile function of isolated left ventricular myocardium from New York Heart Association class IV-failing and nonfailing hearts at physiological temperature and heart rates in order to identify and quantitate abnormalities in myocardial function that underlie abnormal ventricular function. The isometric tension-generating ability of isolated left ventricular strips from nonfailing and failing human hearts was investigated at 37 degrees C and contraction frequencies ranging from 12 to 240 per minute (min-1). Strips were dissected using a new method of protection against cutting injury with 2,3-butanedione monoxime (BDM) as a cardioplegic agent. In nonfailing myocardium the twitch tension-frequency relation is bell-shaped developing 25 +/- 2 mN/mm2 at a contraction frequency of 72 min-1 and peaking at 44 +/- 3.7 mN/mm2 at a contraction frequency of 174 +/- 4 min-1. In failing myocardium the peak of the curve occurs at lower frequencies between 6 and 120 min-1 averaging 81 +/- 22 min-1, and it develops 48% (p less than 0.001) and 80% (p less than 0.001) less tension than in nonfailing myocardium at 72 and 174 min-1, respectively. Between 60 and 150 min-1 tension increases by 107% in nonfailing myocardium, but it does not change significantly in failing myocardium. Peak rates of rise and fall of isometric twitch tension vary in parallel with twitch tension as stimulation frequency rises in nonfailing myocardium but not in failing myocardium. The quantitative agreement between these results from isolated myocardium and those from catheterization laboratory measurements on intact humans suggest that alterations of myocardial origin, independent of systemic factors, may contribute to the above mentioned abnormalities in left ventricular function seen in dilated cardiomyopathy.


Screening for 185delAG in the Ashkenazim.
A study was initiated to assess interest, educational effectiveness, and implications of genetic testing for the common BRCA1 mutation, 185delAG, in the Ashkenazim. Of 333 individuals who attended group sessions, 309 (92%) participated in the study. Participants were categorized as having negative family history (67%), positive family history (defined, by a relaxed criterion, as one first-degree relative or two second-degree relatives with breast [premenopausal] or ovarian cancer) (22%), positive personal history (7%), and both positive personal history and positive family history (4%). Group education was effective, as shown by the improvement in participant scores from pre- to posteducation tests. For the 289 individuals (94%) who requested testing, the major reasons included concern for their own risk, concern for the risk of their children, and desire to learn about surveillance options. The most common reason given by participants who declined testing was concern about health insurance. Six participants found to be heterozygous for the 185delAG mutation received results and were offered genetic counseling. Participants had consented for additional testing without receiving results and were screened for the 6174delT mutation in BRCA2, and seven were found to be positive. All identified carriers reported at least one first- or second-degree relative with a history of breast or ovarian cancer, although they did not all meet our study criteria for positive family history. Given these outcomes, we conclude that screening for breast and ovarian cancer susceptibility is most appropriate for individuals with a positive personal or positive family cancer history. We propose a guideline for future studies designed to identify individuals who may benefit from genetic testing for inherited breast and ovarian cancer.


Host selection in Tomicus piniperda L.: composition of monoterpene hydrocarbons in relation to attack frequency in the shoot feeding phase.
The aim of this study was to investigate the host selection capacity of the pine shoot beetle, Tomicus piniperda, in the shoot-feeding phase and analyze the chiral and non-chiral host volatiles by means of GC-MS and 2D-GC in five Pinus species originating from France (Pinus sylvestris, P. halepensis, P. nigra laricio, P. pinaster maritima, P. pinaster mesogeensis). Dominating monoterpenes were (-)-alpha-pinene, (+)-alpha-pinene, (-)-beta-pinene and (+)-3-carene. The amounts of the enantiomers varied considerably within and among the species. In a principal component analysis-plot, based on the absolute amounts of 18 monoterpene hydrocarbons, separation of the pine species into two groups was obtained. P. halepensis and P. sylvestris were grouped according to the amount of (+)-alpha-pinene and (+)-3-carene, while P. nigra laricio, P. pinaster maritima and P. pinaster mesogeensis were grouped according to (-)-alpha-pinene and (-)-beta-pinene. P. nigra laricio was the species most attacked and P, halepensis the one least attacked by T. piniperda.


Molecular mechanisms underlying resistance to androgen deprivation therapy in prostate cancer.
Prostate cancer (PCa) is the most widely diagnosed male cancer in the Western World and while low- and intermediate-risk PCa patients have a variety of treatment options, metastatic patients are limited to androgen deprivation therapy (ADT). This treatment paradigm has been in place for 75 years due to the unique role of androgens in promoting growth of prostatic epithelial cells via the transcription factor androgen receptor (AR) and downstream signaling pathways. Within 2 to 3 years of ADT, disease recurs-at which time, patients are considered to have castration-recurrent PCa (CR-PCa). A universal mechanism by which PCa becomes resistant to ADT has yet to be discovered. In this review article, we discuss underlying molecular mechanisms by which PCa evades ADT. Several major resistance pathways center on androgen signaling, including intratumoral and adrenal androgen production, AR-overexpression and amplification, expression of AR mutants, and constitutively-active AR splice variants. Other ADT resistance mechanisms, including activation of glucocorticoid receptor and impairment of DNA repair pathways are also discussed. New therapies have been approved for treatment of CR-PCa, but increase median survival by only 2-8 months. We discuss possible mechanisms of resistance to these new ADT agents. Finally, the practicality of the application of "precision oncology" to this continuing challenge of therapy resistance in metastatic or CR-PCa is examined. Empirical validation and clinical-based evidence are definitely needed to prove the superiority of "precision" treatment in providing a more targeted approach and curative therapies over the existing practices that are based on biological "cause-and-effect" relationship.


SecM-stalled ribosomes adopt an altered geometry at the peptidyl transferase center.
As nascent polypeptide chains are synthesized, they pass through a tunnel in the large ribosomal subunit. Interaction between specific nascent chains and the ribosomal tunnel is used to induce translational stalling for the regulation of gene expression. One well-characterized example is the Escherichia coli SecM (secretion monitor) gene product, which induces stalling to up-regulate translation initiation of the downstream secA gene, which is needed for protein export. Although many of the key components of SecM and the ribosomal tunnel have been identified, understanding of the mechanism by which the peptidyl transferase center of the ribosome is inactivated has been lacking. Here we present a cryo-electron microscopy reconstruction of a SecM-stalled ribosome nascent chain complex at 5.6 Å. While no cascade of rRNA conformational changes is evident, this structure reveals the direct interaction between critical residues of SecM and the ribosomal tunnel. Moreover, a shift in the position of the tRNA-nascent peptide linkage of the SecM-tRNA provides a rationale for peptidyl transferase center silencing, conditional on the simultaneous presence of a Pro-tRNA(Pro) in the ribosomal A-site. These results suggest a distinct allosteric mechanism of regulating translational elongation by the SecM stalling peptide.


Targeting Polyamines Inhibits Coronavirus Infection by Reducing Cellular Attachment and Entry.
Coronaviruses first garnered widespread attention in 2002 when the severe acute respiratory syndrome coronavirus (SARS-CoV) emerged from bats in China and rapidly spread in human populations. Since then, Middle East respiratory syndrome coronavirus (MERS-CoV) emerged and still actively infects humans. The recent SARS-CoV-2 outbreak and the resulting disease (coronavirus disease 2019, COVID19) have rapidly and catastrophically spread and highlighted significant limitations to our ability to control and treat infection. Thus, a basic understanding of entry and replication mechanisms of coronaviruses is necessary to rationally evaluate potential antivirals. Here, we show that polyamines, small metabolites synthesized in human cells, facilitate coronavirus replication and the depletion of polyamines with FDA-approved molecules significantly reduces coronavirus replication. We find that diverse coronaviruses, including endemic and epidemic coronaviruses, exhibit reduced attachment and entry into polyamine-depleted cells. We further demonstrate that several molecules targeting the polyamine biosynthetic pathway are antiviral in vitro. In sum, our data suggest that polyamines are critical to coronavirus replication and represent a highly promising drug target in the current and any future coronavirus outbreaks.


Survey of medical services at major league baseball stadiums.
To analyze the availability and level of medical services for fans at major league baseball games in the United States. A 10-item questionnaire was sent to the operations managers of each of 28 major league baseball stadiums. The survey was distributed in cooperation with a major league baseball club. Telephone follow-up was used to complete missing responses. The survey addressed five areas of fan medical services: 1) health-care provider availability and compensation; 2) advanced cardiac life support (ACLS) capabilities, including equipment; 3) presence of on-site ambulance(s); 4) fan fatalities; and 5) alcohol consumption limitations. Survey response was 100%. Healthcare providers are on-site at all stadiums: nurses (86%), physicians (75%), emergency medical technicians (EMTs, [68%]), and paramedics (50%). Ninety-six percent use a combination of health-care providers. The most common medical teams are nurse+EMT+physician (25%) and nurse+EMT+paramedic+physician (18%). All health-care providers receive some form of compensation. All stadiums have at least one ACLS-certified provider; 96% have ACLS equipment. Ambulances are on-site 75% of the time. Sixty-eight percent of the clubs reported at least one fan fatality through the 1992 and 1993 seasons (mean 1.1, range 0-4). All clubs limit alcohol consumption; 96% use multiple approaches. The various approaches include: 1) specific inning discontinuation (86%); 2) maximum purchase (68%); 3) restricted sale locations (64%); and 4) crowd conduct (57%). Advertisement for responsible alcohol consumption is displayed at 75% of the stadiums; designated-driver programs exist at 46%. All major league baseball clubs provide medical services for fans. Furthermore, almost all stadiums have ACLS capabilities. Responsible alcohol consumption also is a recognized priority for fan safety.