Influence of transfer RNA tertiary structure on aminoacylation efficiency by glutaminyl and cysteinyl-tRNA synthetases.
The position of the tertiary Levitt pair between nucleotides 15 and 48 in the transfer RNA core region suggests a key role in stabilizing the joining of the two helical domains, and in maintaining the relative orientations of the D and variable loops. E. coli tRNA(Gln) possesses the canonical Pu15-Py48 trans pairing at this position (G15-C48), while the tRNA(Cys) species from this organism instead features an unusual G15-G48 pair. To explore the structural context dependence of a G15-G48 Levitt pair, a number of tRNA(Gln) species containing G15-G48 were constructed and evaluated as substrates for glutaminyl and cysteinyl-tRNA synthetases. The glutaminylation efficiencies of these mutant tRNAs are reduced by two to tenfold compared with native tRNA(Gln), consistent with previous findings that the tertiary core of this tRNA plays a role in GlnRS recognition. Introduction of tRNA(Cys) identity nucleotides at the acceptor and anticodon ends of tRNA(Gln) produced a tRNA substrate which was efficiently aminoacylated by CysRS, even though the tertiary core region of this species contains the tRNA(Gln) G15-C48 pair. Surprisingly, introduction of G15-G48 into the non-cognate tRNA(Gln) tertiary core then significantly impairs CysRS recognition. By contrast, previous work has shown that CysRS aminoacylates tRNA(Cys) core regions containing G15-G48 with much better efficiency than those with G15-C48. Therefore, tertiary nucleotides surrounding the Levitt pair must significantly modulate the efficiency of aminoacylation by CysRS. To explore the detailed nature of the structural interdependence, crystal structures of two tRNA(Gln) mutants containing G15-G48 were determined bound to GlnRS. These structures show that the larger purine ring of G48 is accommodated by rotation into the syn position, with the N7 nitrogen serving as hydrogen bond acceptor from several groups of G15. The G15-G48 conformations differ significantly compared to that observed in the native tRNA(Cys) structure bound to EF-Tu, further implicating an important role for surrounding nucleotides in maintaining the integrity of the tertiary core and its consequent ability to present crucial recognition determinants to aminoacyl-tRNA synthetases.


The level of physical activity affects adrenal and cardiovascular reactivity to psychosocial stress.
Physical activity plays a key role in the control of neuroendocrine, autonomic, and behavioral responses to physical and psychosocial stress. However, little is known about how the level of physical activity modulates stress responsiveness. Here, we test whether different levels of physical activity are associated with different adrenal, cardiovascular, and psychological responses to psychosocial stress. In addition, competitiveness is assessed as a personality trait that possibly modulates the relationship between physical activity and stress reactivity. Eighteen elite sportsmen, 50 amateur sportsmen, and 24 untrained men were exposed to a standardized psychosocial laboratory stressor (Trier Social Stress Test). Repeated measures of salivary free cortisol, heart rate, and psychological responses to psychosocial stress were compared among the 3 study groups. Elite sportsmen exhibited significantly lower cortisol, heart rate, and state anxiety responses compared with untrained subjects. Amateur sportsmen showed a dissociation between sympathetic and hypothalamic-pituitary-adrenal responsiveness to stress, with significantly reduced heart rate responses but no difference in cortisol responses compared with untrained men. Different levels of competitiveness among groups did not mediate stress reactivity. Our results are in line with previous studies indicating reduced reactivity of the autonomic nervous system to psychosocial stress in trained individuals. More importantly, these findings imply a differential effect of the level of physical activity on different stress-related neurophysiological systems in response to psychosocial stress.


The Golgi complex in living osteoblasts.
Embryo chick leg bones were cultivated under sheets of dialysis cellophane in multipurpose culture chambers. The osteoblasts emigrating in this environment contained phase gray juxtanuclear bodies from which phase white droplets emanated. These were concluded to be the Golgi complex and are described with respect to their living morphology, microscopically observed functional activity, vital staining, and cytochemical reactions.


Direct economic burden of chronic hepatitis C virus in a United States managed care population.
To estimate all-cause and disease-related resource utilization and costs among managed care enrollees with chronic hepatitis C virus (HCV). A large United States claims database was analyzed (1/1/2002 to 12/31/2006). Inclusion criteria were: diagnosis of chronic HCV; no hepatitis B diagnoses; ≥6 and ≥12 months of continuous plan enrollment prediagnosis and postdiagnosis, respectively. Use and costs of medical services and prescription drugs over a 12-month period postdiagnosis were evaluated. Outcomes were assessed in controls without HCV matched (1:1) on age, sex, and plan enrollment. All cost estimates were generated using multivariate generalized linear models to adjust for additional covariates and skewness common in health care cost data. Of the 20,662 patients who met all inclusion criteria, mean age was 49 years; 61% were male. Adjusted all-cause costs were $20,961 per HCV patient, compared with $5451 per control (P<0.0001). Hospitalization occurred in 24% of HCV patients compared with 7% of controls (P<0.0001). Mean inpatient costs were $5892 and $1159 per patient, respectively (P<0.0001). Patients with HCV had higher prescription costs compared with controls ($6191 vs. $1315; P<0.0001). At $6864 per patient, disease-related costs were nearly one-third of all costs in patients with HCV, which exceeded all-cause costs among controls by 26% (P<0.0001). Chronic HCV is a costly disease to managed care organizations. Disease-related costs in HCV exceed all-cause costs in demographically matched controls. Increased efforts in HCV screening and early treatment, particularly before progression to liver cirrhosis, may lead to long-term cost savings in HCV management for managed care systems.


An ambusher's arsenal: chemical crypsis in the puff adder (Bitis arietans).
Ambush foragers use a hunting strategy that places them at risk of predation by both visual and olfaction-oriented predators. Resulting selective pressures have driven the evolution of impressive visual crypsis in many ambushing species, and may have led to the development of chemical crypsis. However, unlike for visual crypsis, few studies have attempted to demonstrate chemical crypsis. Field observations of puff adders (Bitis arietans) going undetected by several scent-orientated predator and prey species led us to investigate chemical crypsis in this ambushing species. We trained dogs (Canis familiaris) and meerkats (Suricata suricatta) to test whether a canid and a herpestid predator could detect B. arietans using olfaction. We also tested for chemical crypsis in five species of active foraging snakes, predicted to be easily detectable. Dogs and meerkats unambiguously indicated active foraging species, but failed to correctly indicate puff adder, confirming that B. arietans employs chemical crypsis. This is the first demonstration of chemical crypsis anti-predatory behaviour, though the phenomenon may be widespread among ambushers, especially those that experience high mortality rates owing to predation. Our study provides additional evidence for the existence of an ongoing chemically mediated arms race between predator and prey species.


Escherichia coli RNase R has dual activities, helicase and RNase.
In Escherichia coli, the cold shock response occurs when there is a temperature downshift from 37 degrees C to 15 degrees C, and this response is characterized by induction of several cold shock proteins, including the DEAD-box helicase CsdA, during the acclimation phase. CsdA is involved in a variety of cellular processes. Our previous studies showed that the helicase activity of CsdA is critical for its function in cold shock acclimation of cells and that the only proteins that were able to complement its function were another helicase, RhlE, an RNA chaperone, CspA, and a cold-inducible exoribonuclease, RNase R. Interestingly, other major 3'-to-5' processing exoribonucleases of E. coli, such as polynucleotide phosphorylase and RNase II, cannot complement the cold shock function of CsdA. Here we carried out a domain analysis of RNase R and showed that this protein has two distinct activities, RNase and helicase, which are independent of each other and are due to different domains. Mutant RNase R proteins that lack the RNase activity but exhibit the helicase activity were able to complement the cold shock function of CsdA, suggesting that only the helicase activity of RNase R is essential for complementation of the cold shock function of CsdA. We also observed that in vivo deletion of the two cold shock domains resulted in a loss of the ability of RNase R to complement the cold shock function of CsdA. We further demonstrated that RNase R exhibits helicase activity in vitro independent of its RNase activity. Our results shed light on the unique properties of RNase R and how it is distinct from other exoribonucleases in E. coli.


Rationale, design and objectives of two phase III, randomised, placebo-controlled studies of GLPG1690, a novel autotaxin inhibitor, in idiopathic pulmonary fibrosis (ISABELA 1 and 2).
While current standard of care (SOC) for idiopathic pulmonary fibrosis (IPF) slows disease progression, prognosis remains poor. Therefore, an unmet need exists for novel, well-tolerated agents that reduce lung function decline and improve quality of life. Here we report the design of two phase III studies of the novel IPF therapy, GLPG1690. Two identically designed, phase III, international, randomised, double-blind, placebo-controlled, parallel-group, multicentre studies (ISABELA 1 and 2) were initiated in November 2018. It is planned that, in each study, 750 subjects with IPF will be randomised 1:1:1 to receive oral GLPG1690 600 mg, GLPG1690 200 mg or placebo, once daily, on top of local SOC, for at least 52 weeks. The primary endpoint is rate of decline of forced vital capacity (FVC) over 52 weeks. Key secondary endpoints are week 52 composite endpoint of disease progression or all-cause mortality (defined as composite endpoint of first occurrence of ≥10% absolute decline in per cent predicted FVC or all-cause mortality at week 52); time to first respiratory-related hospitalisation until end of study; and week 52 change from baseline in the St George's Respiratory Questionnaire total score (a quality-of-life measure). Studies will be conducted in accordance with Good Clinical Practice guidelines, Declaration of Helsinki principles, and local ethical and legal requirements. Results will be reported in a peer-reviewed publication. NCT03711162; NCT03733444.


DNA as a dietary biomarker in Antarctic krill, Euphausia superba.
The diet of Antarctic krill (Euphausia superba) has been studied using a variety of techniques, but current methods still suffer from problems that are difficult to solve. This study examined an alternative approach utilizing DNA as a prey biomarker. Methods were developed for the preservation, extraction, and identification of prey DNA from krill collected in the field. Group-specific polymerase chain reaction (PCR) was used to amplify diatom prey (Phylum: Bacillariophyta) and the results from DNA clone libraries were compared with microscopic diet analysis. DNA analysis was superior to microscopy for prey detection. However, differences in prey relative abundance estimates between the two techniques suggested some bias in the DNA-based estimates. Quantification showed that large amounts of prey DNA had been successfully preserved and extracted. Overall the results suggest that the application of DNA-based diet analysis to krill warrants further investigation, particularly for prey that are difficult to study using other methods.


An assessment of methods for aligning two-dimensional microscope sections to create image volumes.
This study assessed five different methods for aligning microscope images of Nissl-stained sections of mouse brain to form three-dimensional image volumes. Methods exploiting both image content and information from un-sectioned tissue were investigated. The accuracy of reconstruction was estimated using fiducials with known physical properties, demonstrating that methods exploiting tissue content produced distorted image volumes while a method using artificial fiducials produced the most accurate and unbiased alignment. Methodological issues relating to methods of volume reconstruction are discussed and it is recommended that methods using information from un-sectioned tissue be used wherever possible.


FACS purification and transcriptome analysis of drosophila neural stem cells reveals a role for Klumpfuss in self-renewal.
Drosophila neuroblasts (NBs) have emerged as a model for stem cell biology that is ideal for genetic analysis but is limited by the lack of cell-type-specific gene expression data. Here, we describe a method for isolating large numbers of pure NBs and differentiating neurons that retain both cell-cycle and lineage characteristics. We determine transcriptional profiles by mRNA sequencing and identify 28 predicted NB-specific transcription factors that can be arranged in a network containing hubs for Notch signaling, growth control, and chromatin regulation. Overexpression and RNA interference for these factors identify Klumpfuss as a regulator of self-renewal. We show that loss of Klumpfuss function causes premature differentiation and that overexpression results in the formation of transplantable brain tumors. Our data represent a valuable resource for investigating Drosophila developmental neurobiology, and the described method can be applied to other invertebrate stem cell lineages as well.


A citizen science approach to monitoring bleaching in the zoantharian Palythoa tuberculosa.
Coral reef bleaching events are expected to become more frequent and severe in the near future as climate changes. The zoantharian Palythoa tuberculosa bleaches earlier than many scleractinian corals and may serve as an indicator species. Basic monitoring of such species could help to detect and even anticipate bleaching events, especially in areas where more sophisticated approaches that rely on buoy or satellite measurements of sea surface temperature are unavailable or too coarse. One simple and inexpensive monitoring method involves training volunteers to record observations of host color as a proxy for symbiosis quality. Here, we trained university students to take the 'color fingerprint' of a reef by assessing the color of multiple randomly selected colonies of P. tuberculosa at one time point in Okinawa Island, Japan. We tested the reliability of the students' color scores and whether they matched expectations based on previous monthly monitoring of tagged colonies at the same locations. We also measured three traditional metrics of symbiosis quality for comparison: symbiont morphological condition, cell density, and chlorophyll a content. We found that P. tuberculosa color score, although highly correlated among observers, provided little predictive power for the other variables. This was likely due to inherent variation in colony color among generally healthy zoantharians in midwinter, as well as low sample size and brief training owing to the course structure. Despite certain limitations of P. tuberculosa as a focal organism, the citizen science approach to color monitoring has promise, and we outline steps that could improve similar efforts in the future.


Supramolecular curcumin-barium prodrugs for formulating with ceramic particles.
A simple and stable curcumin-ceramic combined formulation was developed with an aim to improve curcumin stability and release profile in the presence of reactive ceramic particles for potential dental and orthopedic applications. For that, curcumin was complexed with barium (Ba(2+)) to prepare curcumin-barium (BaCur) complex. Upon removal of the unbound curcumin and Ba(2+) by dialysis, a water-soluble BaCur complex was obtained. The complex was showing [M+1](+) peak at 10,000-20,000 with multiple fractionation peaks of MALDI-TOF-MS studies, showed that the complex was a supramolecular multimer. The (1)H NMR and FTIR studies revealed that, divalent Ba(2+) interacted predominantly through di-phenolic groups of curcumin to form an end-to-end complex resulted in supramolecular multimer. The overall crystallinity of the BaCur was lower than curcumin as per XRD analysis. The complexation of Ba(2+) to curcumin did not degrade curcumin as per HPLC studies. The fluorescence spectrum was blue shifted upon Ba(2+) complexation with curcumin. Monodisperse nanoparticles with size less than 200dnm was formed, out of the supramolecular complex upon dialysis, as per DLS, and upon loading into pluronic micelles the size was remaining in similar order of magnitude as per DLS and AFM studies. Stability of the curcumin was improved greater than 50% after complexation with Ba(2+) as per UV/Vis spectroscopy. Loading of the supramloecular nanoparticles into pluronic micelles had further improved the stability of curcumin to approx. 70% in water. These BaCur supramolecule nanoparticles can be considered as a new class of prodrugs with improved solubility and stability. Subsequently, ceramic nanoparticles with varying chemical composition were prepared for changing the material surface reactivity in terms of the increase in, degradability, surface pH and protein adsorption. Further, these ceramic particles were combined with curcumin prodrug formulations and optimized the curcumin release properties in the combined formulations. Our proof concept study shows that, the conversion of curcumin to a metal-organic supramolecular prodrug improved the solubility, stability and release profile of curcumin. The prodrug approach with the micellisation strategy appears to be more appropriate to deliver intact curcumin in the presence of ceramic particles of varying surface reactivity.


Use of cannabinoid-based medicine among older residential care recipients diagnosed with dementia: study protocol for a double-blind randomised crossover trial.
Dementia is a neurological condition that affects the cognitive and functional ability of the brain and is the leading cause of disability among those aged 65 years and above. More effective ways to manage dementia symptoms are needed because current treatment options (antidepressants and antipsychotics) can be ineffective and are associated with substantial side effects, including increased rate of mortality. Cannabinoid-based medicine (CBM) has shown an ability to inhibit some symptoms associated with dementia, and the adverse effects are often minimal; yet, little research has explored the use of CBM among this population. To monitor the safety of a purified dose of CBM oil (3:2 delta-9-tetrahydrocannabinol:cannabidiol) on behaviour symptoms, quality of life and discomfort caused by pain. We will carry out an 18-week, randomised, double-blind crossover trial that consists of a 2-week eligibility period, two 6-week treatment cycles, and two 2-week washout periods (between both cycles and after the second treatment cycle). We aim to recruit 50 participants with dementia who are living in residential aged-care facilities. The participants will be randomised into two groups and will receive a dose of either CBM oil or placebo for the first treatment cycle and the opposite medication for the second. Data will be collected using the Neuropsychiatric Inventory Questionnaire, the Cohen-Mansfield Agitation Inventory, the Quality of Life in Alzheimer's Disease questionnaire, and the Abbey Pain Scale on seven occasions. These will be completed by the participants, aged-care staff, and nominated next of kin or family members. The participants' heart rate and blood pressure will be monitored weekly, and their body composition and weight will be monitored fortnightly by a research nurse, to assess individual dose response and frailty. In addition, pre- and post-surveys will be administered to aged-care staff and family members to understand their perceptions of CBM and to inform proposed focus groups consisting of the aged-care staff and next of kin. The study design has been informed by medical professionals and key stakeholders, including those working in the residential aged-care industry to ensure patient safety, collection of non-invasive measures, and methodological rigor and study feasibility. Australian New Zealand Clinical Trials Registry, ACTRN12619000474156. Registered on 21 March 2019.


ETS transcription factors in hematopoietic stem cell development.
Hematopoietic stem cells (HSCs) are essential for the maintenance of the hematopoietic system. However, these cells cannot be maintained or created in vitro, and very little is known about their generation during embryogenesis. Many transcription factors and signaling pathways play essential roles at various stages of HSC development. Members of the ETS ('E twenty-six') family of transcription factors are recognized as key regulators within the gene regulatory networks governing hematopoiesis, including the ontogeny of HSCs. Remarkably, although all ETS transcription factors bind the same DNA consensus sequence and overlapping tissue expression is observed, individual ETS transcription factors play unique roles in the development of HSCs. Also, these transcription factors are recurrently used throughout development and their functions are context-dependent, increasing the challenge of studying their mechanism of action. Critically, ETS factors also play roles under pathological conditions, such as leukemia and, therefore, deciphering their mechanism of action will not only enhance our knowledge of normal hematopoiesis, but also inform protocols for their creation in vitro from pluripotent stem cells and the design of new therapeutic approaches for the treatment of malignant blood cell diseases. In this review, we summarize the key findings on the roles of ETS transcription factors in HSC development and discuss novel mechanisms by which they could control hematopoiesis.


A self optimizing synthetic organic reactor system using real-time in-line NMR spectroscopy.
A configurable platform for synthetic chemistry incorporating an in-line benchtop NMR that is capable of monitoring and controlling organic reactions in real-time is presented. The platform is controlled via a modular LabView software control system for the hardware, NMR, data analysis and feedback optimization. Using this platform we report the real-time advanced structural characterization of reaction mixtures, including 19F, 13C, DEPT, 2D NMR spectroscopy (COSY, HSQC and 19F-COSY) for the first time. Finally, the potential of this technique is demonstrated through the optimization of a catalytic organic reaction in real-time, showing its applicability to self-optimizing systems using criteria such as stereoselectivity, multi-nuclear measurements or 2D correlations.


Iron oxide nanoparticles induce Pseudomonas aeruginosa growth, induce biofilm formation, and inhibit antimicrobial peptide function.
Given the increased use of iron-containing nanoparticles in a number of applications, it is important to understand any effects that iron-containing nanoparticles can have on the environment and human health. Since iron concentrations are extremely low in body fluids, there is potential that iron-containing nanoparticles may influence the ability of bacteria to scavenge iron for growth, affect virulence and inhibit antimicrobial peptide (AMP) function. In this study, Pseudomonas aeruginosa (PA01) and AMPs were exposed to iron oxide nanoparticles, hematite (α-Fe2O3), of different sizes ranging from 2 to 540 nm (2 ± 1, 43 ± 6, 85 ± 25 and 540 ± 90 nm) in diameter. Here we show that the greatest effect on bacterial growth, biofilm formation, and AMP function impairment is found when exposed to the smallest particles. These results are attributed in large part to enhanced dissolution observed for the smallest particles and an increase in the amount of bioavailable iron. Furthermore, AMP function can be additionally impaired by adsorption onto nanoparticle surfaces. In particular, lysozyme readily adsorbs onto the nanoparticle surface which can lead to loss of peptide activity. Thus, this current study shows that co-exposure of nanoparticles and known pathogens can impact host innate immunity. Therefore, it is important that future studies be designed to further understand these types of impacts.


Felodipine-diazabicyclo[2.2.2]octane-water (1/1/1).
The title compound, C(18)H(19)Cl(2)NO(4)·C(6)H(12)N(2)·H(2)O, is a cocrystal hydrate containing the active pharmaceutical ingredient felodipine and diazabicyclo[2.2.2]octane (DABCO). The DABCO and water molecules are linked through O-H···N hydrogen bonds into chains around 2(1) screw axes, while the felodipine molecules form N-H···O hydrogen bonds to the water molecules. The felodipine molecules adopt centrosymmetric back-to-back arrangements that are similar to those present in all of its four reported polymorphs. The dichlorophenyl rings also form π-stacking interactions. The inclusion of water molecules in the cocrystal, rather than formation of N-H···N hydrogen bonds between felodipine and DABCO, may be associated with steric hindrance that would arise between DABCO and the methyl groups of felodipine if they were directly involved in hydrogen bonding.


Practical management: vocal cord dysfunction in athletes.
Vocal cord dysfunction (VCD) is characterized by paradoxical adduction of the vocal folds during inhalation, and occasionally upon exhalation, resulting in extrathoracic airflow obstruction. Sports medicine professionals must have a high index of suspicion for VCD when acute respiratory symptoms occur so that prompt evaluation and use of appropriate specialists results in an accurate and timely diagnosis. Many factors have been implicated in the pathophysiology of VCD, including laryngeal irritants, psychogenic and neurogenic causes. The diagnosis and management of VCD involves a variety of specialties including pulmonology, otolaryngology, speech-language pathology, allergy and immunology, and psychologic management as appropriate. The mainstay of treatment remains behavioral management guided by a medical speech-language pathologist, as well as pharmacologic management for VCD triggers.


Simple microfluidic device for detecting the negative dielectrophoresis of DNA labeled microbeads.
We propose a new microfluidic device that can be used to determine the change in the negative dielectrophoresis (n-DEP) of dielectric microbeads when a small amount of DNA is attached to them. We previously proposed a DNA detection method based on changes in the DEP of microbeads induced by the attachment of DNA. When target DNA is attached to the microbeads having n-DEP property, the DEP changes from negative to positive. This occurs because electric charges of the DNA increase the surface conductance of the microbeads. Thus, only the DNA-labeled microbeads are attracted to a microelectrode by positive DEP. The trapped DNA-labeled microbeads can be counted by dielectrophoretic impedance measurements. A large amount of DNA (approximately 105 DNA molecules) is required to change the DEP from negative to positive. Even though this method can be combined with DNA amplification, reducing the amount of DNA required can help us to shorten the reaction time. In this study, we aimed to detect DNA less than 105 DNA molecules by determining the change in the n-DEP change. To achieve this, we proposed a simple microfluidic device consisting of a single microchannel and a single pair of microelectrodes. Numerical simulations revealed that the device can identify the slight change in the n-DEP of the microbeads corresponding to the attachment of a small amount of DNA. In practical experiments, the fabricated device distinguished 10-1000 DNA molecules per microbead. This method represents a fast and easy method of DNA detection when combined with DNA amplification techniques.


First Report of Petunia Root Rot Caused by Rhizoctonia solani in Argentina.
Common garden petunias (Petunia × hybrida Hort. Vilm.-Andr.) are herbaceous annual plants with brightly colored flowers up to 10 cm in diameter. During the winter of 2002, crown and root rot were observed on plants (cv. Ultra) growing in five greenhouses in Buenos Aires. Affected plants were randomly distributed in the greenhouses, and mean disease incidence in all the greenhouses was 26%. Basal leaves turned yellow and gradually became necrotic, and infected plants were often killed. Small pieces of affected tissues were disinfested in 2% sodium hypochlorite for 1 min and plated on 2% potato dextrose agar (PDA). Fifteen isolates identified to the genus Rhizoctonia were obtained. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Hyphal branched at right angles, were constricted at the base of the branch near the union with main hyphae, and septate near the constriction. Basidia were not observed in the greenhouses or on the plates. Isolates were cultivated on water agar and incubated at 25°C for 3 days. Hyphal cells were determined to be multinucleate when stained with 1% aniline blue solution (2) and examined at ×400. Anastomosis group of one isolate was determined by using AG-4 HG II, AG-1 IA, AG-1 IB, AG-1 IC, AG-2 2-1, and AG-2 2IIIB tester strains of Rhizoctonia solani that includes isolates reported to be pathogenic on ornamentals (1). Anastomosis was observed only with strains of AG-4 HG II. Pathogenicity on this isolate was conducted on potted, healthy, adult plants that were 10 to 22 cm high and flowering. Thirty-five plants were inoculated by placing 1 cm2 pieces of PDA from 7-day-old mycelial cultures near the base of the stem. Twelve control plants were treated with 1 cm2 PDA plugs. Plants were kept at 22 to 24°C, >95% relative humidity, and 12 h of fluorescent light. Wilt symptoms due to basal stem rot appeared 7 days after inoculation, and all the inoculated plants died within 27 days. Control plants remained disease free. The pathogen was reisolated from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report of R. solani causing disease on petunia in Argentina. References: (1) D. M. Benson and D. K. Cartwright. Ornamental diseases incited by Rhizoctonia spp. Pages 303-314 in: Rhizoctonia species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, London, England, 1996. (2) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.


A Fault Diagnosis Methodology for Gear Pump Based on EEMD and Bayesian Network.
This paper proposes a fault diagnosis methodology for a gear pump based on the ensemble empirical mode decomposition (EEMD) method and the Bayesian network. Essentially, the presented scheme is a multi-source information fusion based methodology. Compared with the conventional fault diagnosis with only EEMD, the proposed method is able to take advantage of all useful information besides sensor signals. The presented diagnostic Bayesian network consists of a fault layer, a fault feature layer and a multi-source information layer. Vibration signals from sensor measurement are decomposed by the EEMD method and the energy of intrinsic mode functions (IMFs) are calculated as fault features. These features are added into the fault feature layer in the Bayesian network. The other sources of useful information are added to the information layer. The generalized three-layer Bayesian network can be developed by fully incorporating faults and fault symptoms as well as other useful information such as naked eye inspection and maintenance records. Therefore, diagnostic accuracy and capacity can be improved. The proposed methodology is applied to the fault diagnosis of a gear pump and the structure and parameters of the Bayesian network is established. Compared with artificial neural network and support vector machine classification algorithms, the proposed model has the best diagnostic performance when sensor data is used only. A case study has demonstrated that some information from human observation or system repair records is very helpful to the fault diagnosis. It is effective and efficient in diagnosing faults based on uncertain, incomplete information.


The changing clinical pattern of Reye's syndrome 1982-1990.
To describe trends in the clinical pattern of Reye's syndrome in the British Isles between 1982 and 1990; and to determine the relation between any changes and the June 1986 warnings against the use of aspirin in children. Development, and application to reported cases, of a scoring system designed such that patients showing the typical clinical and pathological features of 'classical' Reye's syndrome scored highly. The relations between 'Reye scores' and a number of explanatory variables were explored using multivariable analysis. British Isles. 445 cases fulfilling the Reye's syndrome case definition reported to the surveillance scheme between January 1982 and December 1990. Individual 'Reye score'. Cases with high scores were more likely to have occurred in the 4 1/2 year period before June 1986 compared with the subsequent period (p < 0.006). Numbers of cases in the low and intermediate score categories declined by about 50% after June 1986, whereas those in the high category fell by 79%. High scorers were more likely to have received aspirin (p < 0.0001) and were older than intermediate and low scorers (p < 0.008). No relation was identified between score and season of onset. The decline in Reye's syndrome after the aspirin warnings cannot be explained entirely, as has been proposed, by improved diagnosis of 'Reye-like' inherited metabolic and other disorders: this would not account for the greater decline of the high scoring subgroup which also contained those cases most likely to resemble 'classical' Reye's syndrome and to have received aspirin. This study provides further evidence for the role of aspirin in a subset of cases meeting the standard diagnostic criteria for Reye's syndrome and supports the need to consider this disorder as a heterogeneous group of conditions including Reye-like inherited metabolic disorders.


The Heterodimeric ABC Transporter EfrCD Mediates Multidrug Efflux in Enterococcus faecalis.
Nosocomial infections with Enterococcus faecalis are an emerging health problem. However, drug efflux pumps contributing to intrinsic drug resistance are poorly studied in this Gram-positive pathogen. In this study, we functionally investigated seven heterodimeric ABC transporters of E. faecalis that are annotated as drug efflux pumps. Deletion of ef0789-ef0790 on the chromosome of E. faecalis resulted in increased susceptibility to daunorubicin, doxorubicin, ethidium, and Hoechst 33342, and the corresponding transporter was named EfrCD. Unexpectedly, the previously described heterodimeric multidrug ABC transporter EfrAB contributes marginally to drug efflux in the endogenous context of E. faecalis In contrast, heterologous expression in Lactococcus lactis revealed that EfrAB, EfrCD, and the product of ef2226-ef2227 (EfrEF) mediate the efflux of fluorescent substrates and confer resistance to multiple dyes and drugs, including fluoroquinolones. Four of seven transporters failed to exhibit drug efflux activity for the set of drugs and dyes tested, even upon overexpression in L. lactis Since all seven transporters were purified as heterodimers after overexpression in L. lactis, a lack of drug efflux activity is not attributed to poor expression or protein aggregation. Reconstitution of the purified multidrug transporters EfrAB, EfrCD, and EfrEF in proteoliposomes revealed functional coupling between ATP hydrolysis and drug binding. Our analysis creates an experimental basis for the accurate prediction of drug efflux transporters and indicates that many annotated multidrug efflux pumps might be incapable of drug transport and thus might fulfill other physiological functions in the cell.


The Surgical Choice of Incidental Periampullary Carcinoma Management in Emergency Laparotomy.
BACKGROUND Laparotomy patients are occasionally diagnosed as having incidental periampullary cancers, making emergency pancreaticoduodenectomy (PD) inevitable. In this situation is difficult to decide whether to perform an emergency PD or a two-stage PD. MATERIAL AND METHODS A total of 27 patients who underwent emergency abdominal laparotomy were diagnosed with periampullary or pancreatic cancer during the operation without enough preoperative preparation. Ten patients underwent emergency one-stage PD and 17 patients underwent two-stage PD. Data of 137 patients with elective PD were selected as the control group. The preoperative, operative, and postoperative parameters, including hospital stay, medical cost, blood loss, and postoperative complications between elective PD and emergency PD (one-stage and two-stage) and between one-stage PD and two-stage PD were analyzed by chi-square test, Fisher test, or t test. RESULTS Patients undergoing emergency two-stage PD had less blood loss (P=0.014), while patients with one-stage PD had shorter hospital stay (P=0.004), shorter operation time (P=0.047), and lower treatment costs (P=0.003). Additionally, the complications rates between one-stage and two-stage PD had no significant difference (P=0.365). Elective PD was the optimal method due to shorter hospital stay (P<0.001), less hemorrhage (P<0.001), shorter operative time (P<0.001), and lower cost (P<0.001) compared with emergency PD. CONCLUSIONS Based on our experience, one-stage PD had advantages of shorter hospital stay, shorter operation time, and lower treatment costs, while two-stage PD had less blood loss. The emergency two-stage PD may be more suitable for patients with unstable vital signs if emergency PD is inevitable in an emergency laparotomy.


Insulin Resistance is Associated with Central Pain in Patients with Fibromyalgia.
Insulin resistance (IR) is a pathological condition in which cells fail to respond normally to insulin. IR has been associated with multiple conditions, including chronic pain. Fibromyalgia (FM) is one of the common generalized chronic painful conditions with an incidence rate affecting 3% to 6% of the population. Substantial interest and investigation into FM continue to generate many hypotheses.The relationship between IR and FM has not been explored. IR is known to cause abnormalities in the cerebral microvasculature, leading to focal hypoperfusion. IR also has been shown to cause cognitive impairment in FM patients, as in parkinsonism. As demonstrated by advanced imaging methods, similar brain perfusion abnormalities occur in the brain of patients with FM as with IR. To determine the potential association between FM and IR. Subspecialty pain medicine clinics. Observational cross-sectional study. Laboratory data was extracted through a retrospective review of medical records from patients who had met the American College of Rheumatology (ACR) criteria for FM. The Hemoglobin A1c (HbA1c) values from 33 patients with FM were compared with the means of the glycated HbA1c levels of 2 control populations. In addition, established indices of IR [Quantitative Insulin Sensitivity Check Index (QUICKI) and the Homeostatic Model Assessment of Insulin Resistance (HOMA-IR)] were calculated in a subgroup of patients in whom the analytes necessary for these calculations were available. To assess for confounding factors, the associations between HbA1c, QUICKI, HOMA-IR, fasting insulin levels, and glucose, after controlling for age, were explored by multiple analyses of variance with relation to gender and ethnicity. We found an association between IR and FM that was independent of age, gender, and ethnicity. We found that patients with FM belong to a distinct population that can be segregated from the control groups by their HbA1c levels, a surrogate marker of IR. This was demonstrated by analyzing the data after introducing an age correction into a linear regression model. This strategy showed significant differences between patients with FM and control subjects (P < 0.0001 and P = 0.0002, for 2 separate control populations, respectively). A subgroup analysis using the QUICKI and HOMA-IR showed that all patients with FM in this subgroup (100%) exhibited laboratory abnormalities pointing to IR. Small observational cross-sectional study. There are also intrinsic limitations that are attributed to cross-sectional studies. The association demonstrated in this study warrant further investigation, including the pursuit of randomized, double-blind clinical trials to determine the effect of improving insulin sensitivity in FM related pain scores. Such studies could unveil a potential pathogenetic relationship between FM, central pain, and IR. Based on these initial findings, we present the hypothesis that IR may underlie pathological mechanisms leading to central pain. If confirmed, this may lead to a paradigm shift in the management of central pain.


Full and partial peroxisome proliferation-activated receptor-γ agonists, but not δ agonist, rescue of dopaminergic neurons in the 6-OHDA parkinsonian model is associated with inhibition of microglial activation and MMP expression.
Peroxisome proliferator-activated receptor gamma (PPARγ) is a nuclear hormone receptor that has been shown to have anti-inflammatory and matrix metalloproteinase (MMP) inhibitor properties. PPARγ agonists have been shown to have neuroprotective effects in various neurodegeneration models where inflammation is implicated, including models of Parkinson's disease. However, no studies have looked at the effects of partial PPARγ agonists. The neuroprotective effects of the PPARγ full agonist, pioglitazone (20 mg/kg), partial PPARγ agonist GW855266X (15 mg/kg) and PPAR-δ full agonist GW610742X (10 mg/kg) were investigated in the 6-hydroxydopamine (6-OHDA) model of Parkinson's disease when administered prior to or post 6-OHDA lesioning. The integrity of the nigrostriatal system was assessed by assessing the numbers dopaminergic neurons in the substantia nigra (SN) and by assessing striatal dopamine content. The degree of microglia activation in the SN was also immunohistochemistry assessed utilizing the marker OX-6 for activated microglia and CD-68 a marker for phagocytic microglia. Additionally we performed immunocytochemistry for MMP3 in the SN. Finally, we investigated whether a period of drug withdrawal for a further 7 days affected the neuroprotection produced by the PPARγ agonists. Both pioglitazone and GW855266X protected against 6-OHDA induced loss of dopaminergic neurons in the substantia nigra and depletion of striatal dopamine when administered orally twice daily for either 1) 7 day prior to and 7 days post lesioning or 2) for 7 days starting 2 days post lesioning when neurons will be severely traumatized. 6-OHDA lesioning was associated with an increase in microglia activation and in numbers of MMP-3 immunoreactive cells which was attenuated by pioglitazone and GW855266X. Neuroprotective effects were not replicated using the PPARδ agonist GW610742X. Subsequent withdrawal of both pioglitazone and GW855266X, for a further 7 days negated any neuroprotective effect suggesting that long-term administration may be required to attenuate the inflammatory response. For the first time a partial PPAR-γ agonist has been shown to be neuroprotectory when administered post lesioning in a parkinsonian model. Effects may be via the inhibition of microglial and MMP activation and support further research.


Variation of community consultation and public disclosure for a pediatric multi-centered "Exception from Informed Consent" trial.
The US federal regulation "Exception from Informed Consent for Emergency Research," 21 Code of Federal Regulations 50.24, permits emergency research without informed consent under limited conditions. Additional safeguards to protect human subjects include requirements for community consultation and public disclosure prior to starting the research. Because the regulations are vague about these requirements, Institutional Review Boards determine the adequacy of these activities at a local level. Thus, there is potential for broad interpretation and practice variation. To describe the variation of community consultation and public disclosure activities approved by Institutional Review Boards, and the effectiveness of this process for a multi-center, Exception from Informed Consent, pediatric status epilepticus clinical research trial. Community consultation and public disclosure activities were analyzed for each of the 15 participating sites. Surveys were conducted with participants enrolled in the status epilepticus trial to assess the effectiveness of public disclosure dissemination prior to study enrollment. Every Institutional Review Board, among the 15 participating sites, had a varied interpretation of Exception from Informed Consent regulations for community consultation and public disclosure activities. Institutional Review Boards required various combinations of focus groups, interviews, surveys, and meetings for community consultation, and news releases, mailings, and public service announcements for public disclosure. At least 4335 patients received information about the study from these efforts. In all, 158 chose to be included in the "Opt Out" list. Of the 304 participants who were enrolled under Exception from Informed Consent, 12 (5%) had heard about the study through community consultation or public disclosure activities. The activities reaching the highest number of participants were surveys and focus groups associated with existing meetings. Public disclosure activities were more efficient and cost-effective if they were part of an in-hospital resource for patients and families. There is substantial variation in Institutional Review Boards' interpretations of the federal regulations for community consultation and public disclosure. One of the goals of community consultation and public disclosure efforts for emergency research is to provide community members an opportunity to opt out of Exception from Informed Consent research; however, rarely do patients or their legally authorized representatives report having learned about a study prior to enrollment.


Development of a flow method for the hydroboration/oxidation of olefins.
A method for the continuous preparation of alcohols by hydroboration/oxidation of olefins using flow techniques is described. The process allows the isolation of up to 120 mmol h(-1) of the desired alcohol in a very rapid manner with good functional group tolerance. The flow setup can be modified to perform a continuous extraction of the desired alcohol from the biphasic mixture produced by the reaction.


Role of hepatic PKCβ in nutritional regulation of hepatic glycogen synthesis.
The signaling mechanisms by which dietary fat and cholesterol signals regulate central pathways of glucose homeostasis are not completely understood. By using a hepatocyte-specific PKCβ-deficient (PKCβHep-/-) mouse model, we demonstrated the role of hepatic PKCβ in slowing disposal of glucose overload by suppressing glycogenesis and increasing hepatic glucose output. PKCβHep-/- mice exhibited lower plasma glucose under the fed condition, modestly improved systemic glucose tolerance and mildly suppressed gluconeogenesis, increased hepatic glycogen accumulation and synthesis due to elevated glucokinase expression and activated glycogen synthase (GS), and suppressed glucose-6-phosphatase expression compared with controls. These events were independent of hepatic AKT/GSK-3α/β signaling and were accompanied by increased HNF-4α transactivation, reduced FoxO1 protein abundance, and elevated expression of GS targeting protein phosphatase 1 regulatory subunit 3C in the PKCβHep-/- liver compared with controls. The above data strongly imply that hepatic PKCβ deficiency causes hypoglycemia postprandially by promoting glucose phosphorylation via upregulating glucokinase and subsequently redirecting more glucose-6-phosphate to glycogen via activating GS. In summary, hepatic PKCβ has a unique and essential ability to induce a coordinated response that negatively affects glycogenesis at multiple levels under physiological postprandial conditions, thereby integrating nutritional fat intake with dysregulation of glucose homeostasis.


Psychosomatic Associations Between Thinking Patterns and Parkinson's Disease from a Yoga Philosophy Perspective: A New Zealand Cross-Sectional Study.
When investigating the etiology of diseases, epidemiological observational studies traditionally deemphasize psychosomatic associations. Exploring cognitive behavior provides an insight into how psychosomatic associations affect disease. Yoga philosophy identifies the kleshas (mental afflictions) of ignorance, ego, desire, hatred, and fear of death with disease. This is because individuals' perceptions and beliefs generate and reflect streams of thought that may shape their behavior and manifest as, or predispose them to, particular disease(s). The present study takes a yogic philosophical perspective to help elucidate unexplored associations between thinking about different aspects of life and the severity of Parkinson's disease (PD). The study involved a cross-sectional sample survey. Parkinson's New Zealand selected a random sample of 990 of its members. A self-completed questionnaire was sent to them. It asked questions about how often, over the previous 4 weeks, they had thought about 18 aspects of life commonly associated with the kleshas. A completed questionnaire was returned by 319 people (32%). Respondents thought most about family (87%), health (64%), rest and sleep (57%), food (53%), and the future (52%). They reported thinking least about work (48%), sex (45%), death (42%), and being virtuous (39%). A weak, but hypothesized positive, association (r = 0.2, p < 0.000) was detected between PD severity and thinking about death. The study could not determine whether thinking about death was (1) a cause or consequence of PD severity, and (2) a premorbid behavior. However, the possibility that stress associated with thinking about death accelerates PD progression is consistent with yoga philosophy and with neurophysiological mechanisms associated with the psychosomatic connections. The findings are worthy of future testing. A retrospective cohort study and qualitative research could deepen understanding about the role of kleshas in PD.


A transnational perspective of global and regional ecosystem service flows from and to mountain regions.
The spatial relationships of ecosystem services are complex and poorly understood due to spatial mismatches between areas of provision and the areas that benefit. In this study, we assess the spatial flows of six key ecosystem services from and to mountain regions at the regional and global level. We identify major directions of spatial flow and illustrate different types and transfer mechanisms with detailed examples focusing on the European Alps and surrounding lowlands. Our results demonstrate that the spatial flows of ecosystem services range from local to global interactions and extend far beyond the regional level for most of the ecosystem services assessed. Transportation processes encompass passive biophysical processes and the active transportation of goods, distribution of information and traveling of people. Decision and policy-making can use this enhanced understanding to influence ecosystem service transfer and consequently manage natural resources in a sustainable way.


Bacterial keratitis: predisposing factors, clinical and microbiological review of 300 cases.
To identify predisposing factors and to define clinical and microbiological characteristics of bacterial keratitis in current practice. A retrospective analysis of the hospital records of patients presenting with bacterial keratitis and treated at the Quinze-Vingts National Center of Ophthalmology, Paris, France, was performed during a 20 month period. A bacterial keratitis was defined as a suppurative corneal infiltrate and overlying epithelial defect associated with presence of bacteria on corneal scraping and/or that was cured with antibiotic therapy. Risk factors, clinical and microbiological data were collected. 300 cases (291 patients) of presumed bacterial keratitis were included. Potential predisposing factors, usually multiple, were identified in 90.6% of cases. Contact lens wear was the main risk factor (50.3%). Trauma or a history of keratopathy was found in 15% and 21% of cases, respectively. An organism was identified in 201 eyes (68%). 83% of the infections involved Gram positive bacteria, 17% involved Gram negative bacteria, and 2% were polymicrobial. Gram negative bacteria were associated with severe anterior chamber inflammation (p=0.004), as well as greater surface of infiltrates (p=0.01). 99% of ulcers resolved with treatment, but only 60% of patients had visual acuity better than the level at admission, and 5% had very poor visual outcome. Contact lens wear is the most important risk factor. Most community acquired bacterial ulcers resolve with appropriate treatment.


Low hepatitis B vaccine uptake among surgical residents in Cameroon.
Hepatitis B virus (HBV) infection is one of the most serious occupational hazards faced by healthcare workers. Surgical personnel are particularly at risk. HBV infection is preventable by vaccination, but no previous study has assessed HBV vaccination coverage among healthcare workers in Cameroon. We assessed knowledge of risk factors of HBV infection, awareness of HBV vaccine, and vaccination status of surgical residents in Cameroon. A structured pretested questionnaire was administered to 49 of the 70 surgical residents in Cameroon during the 2011-2012 academic year. Since the beginning of their residency program, 28 (57.1%) had had at least one accidental exposure to blood, with a median of 2 (range 1 to 25) exposures. Most of them had a good knowledge of risk factors for HBV infection. Although 98.0% (n = 48) were aware of the HBV vaccine, and 89.8% (n = 44) knew that they were at high risk of infection, only 24.5% (n = 12) had received a full course of at least three doses of the vaccine. In addition, only 33.3% (4/12) underwent post-vaccination testing to confirm a good immunological response (and thus effective protection against HBV infection). Among the 53.1% (n = 28) who had never had any dose of HBV vaccine, the main reasons for not being vaccinated were lack of time (38.5%), lack of money to pay for vaccine (23.1%), and lack of sufficient information on the vaccine (19.2%). Only 20.4% (n = 10) had been sensitized by their training institutions about the importance of HBV vaccination. There is a low HBV vaccine uptake among surgical residents in Cameroon. As part of occupational safety measures, complete HBV vaccination should be strongly recommended and offered to surgical trainees before the beginning of their training program.


Evidence for a direct growth-stimulating effect of estradiol on human MCF-7 cells in vivo.
The MCF-7 continuous line of human breast cancer cells requires that athymic nude mice receive supplemental estrogen so that inocula can produce progressively growing tumors. Although these cells contain a typical estrogen receptor complex, the lack of consistent growth stimulation induced by estrogens added to in vitro culture systems has raised the question as to whether this class of hormones acts directly upon the cells or induces a second message produced in other tissues. The present experiments were designed to test the effect of estradiol on the growth of these cells in vivo by exposing them directly to the hormone prior to its absorption into the hepatic portal circulation and subsequent metabolic inactivation. Tumor fragments that were placed next to an estradiol-containing pellet in the spleen grew to produce grossly evident tumor masses, whereas those in the subcutis of the same animals did not, although some minute residua did remain. In the splenic tumors, the mitotic index of the MCF-7 cells immediately adjacent to the estrogen pellets was 2.4 times that of cells on the other side of the same tumor and 3.5 times that of those in the minute s.c. residua. We interpret these data as indicating that in vivo estradiol is acting directly upon the MCF-7 cells to increase their rate of proliferation rather than to initiate the production of a second message to be released into the circulation. Additionally, it was found that s.c. tumors that were decreasing in volume subsequent to withdrawal of systemic estrogen still contained dividing neoplastic cells but with a lower frequency than that seen in progressively growting tumors stimulated with estradiol. This finding indicates that MCF-7 cells can proliferate in vivo in the absence of a substantial amount of estrogen but only at a rate insufficient to sustain progressive tumor growth.


Vibrational energy flow controls internal conversion in a transition metal complex.
Internal conversion (IC) between excited electronic states is a fundamental photophysical process that is important for understanding protection from UV radiation, energy transfer pathways and electron injection in artificial photosynthetic systems and organic solar cells. We have studied IC between three singlet MLCT states in an iridium complex using femtosecond fluorescence spectroscopy. Very fast IC with a time constant of <20 fs is observed from the highest state and a much slower relaxation to the lowest energy singlet state on a 70 fs time scale. The abrupt slowdown of the relaxation rate occurs when there is >0.6 eV of vibrational energy stored in the complex that has to be dissipated by intramolecular vibrational redistribution before further IC to the lower energy states can occur. These results show that the ability to dissipate vibrational energy can control the relaxation process in this class of materials.


Bartonella species and trombiculid mites of rats from the Mekong Delta of Vietnam.
A survey of Bartonella spp. from 275 rats purchased in food markets (n=150) and trapped in different ecosystems (rice field, forest, and animal farms) (n=125) was carried out during October, 2012-March, 2013, in the Mekong Delta of Vietnam. The overall Bartonella spp. prevalence detected by culture and PCR in blood was 14.9% (10.7-19.1%), the highest corresponding to Rattus tanezumi (49.2%), followed by Rattus norvegicus (20.7%). Trapped rats were also investigated for the presence and type of chiggers (larvae of trombiculid mites), and Bartonella spp. were investigated on chigger pools collected from each rat by RT-PCR. A total of five Bartonella spp. were identified in rats, three of which (B. elizabethae, B. rattimassiliensis, and B. tribocorum) are known zoonotic pathogens. Among trapped rats, factors independently associated with increased prevalence of Bartonella spp. included: (1) Rat species (R. tanezumi); (2) the number of Trombiculini-Blankaartia and Schoengastiini-Ascoschoengastia mites found on rats; and (3) the habitat of the rat (i.e., forest/fields vs. animal farms). The prevalence of Bartonella infection among chiggers from Bartonella spp.-positive R. tanezumi rats was 5/25 (25%), compared with 1/27 (3.7%) among Bartonella spp.-negative R. tanezumi rats (relative risk [RR]=5.4, 95% confidence interval [CI] 0.68-43.09). The finding of Bartonella spp.-positive chiggers on Bartonella spp.-negative rats is strongly suggestive of a transovarial transmission cycle. Rats are ubiquitous in areas of human activity and farms in the Mekong Delta; in addition, trapping and trading of rats for food is common. To correctly assess the human risks due to rat trapping, marketing, and carcass dressing, further studies are needed to establish the routes of transmission and cycle of infection. The widespread presence of these zoonotic pathogens in rats and the abundance of human-rat interactions suggest that surveillance efforts should be enhanced to detect any human cases of Bartonella infection that may arise.


Induction of human immunodeficiency virus type 1-specific T cells by a bluetongue virus tubule-vectored vaccine prime-recombinant modified virus Ankara boost regimen.
In the absence of strategies for reliable induction of antibodies broadly neutralizing human immunodeficiency virus type 1 (HIV-1), vaccine efforts have shifted toward the induction of cell-mediated immunity. Here we describe the construction and immunogenicity of novel T-cell vaccine NS1.HIVA, which delivers the HIV-1 clade A consensus-derived immunogen HIVA on the surface of tubular structures spontaneously formed by protein NS1 of bluetongue virus. We demonstrated that NS1 tubules can accommodate a protein as large as 527 amino acids without losing their self-assembly capability. When injected into BALB/c mice by several routes, chimeric NS1.HIVA tubules induced HIV-1-specific major histocompatibility complex class I-restricted T cells. These could be boosted by modified virus Ankara expressing the same immunogen and generate a memory capable of gamma interferon (IFN-gamma) production, proliferation, and lysis of sensitized target cells. Induced memory T cells readily produced IFN-gamma 230 days postimmunization, and upon a surrogate virus challenge, NS1.HIVA vaccine alone decreased the vaccinia virus vv.HIVA load in ovaries by 2 orders of magnitude 280 days after immunization. Thus, because of its T-cell immunogenicity and antigenic simplicity, the NS1 delivery system could serve as a priming agent for heterologous prime-boost vaccination regimens. Its usefulness in primates, including humans, remains to be determined.


Young adults and the Affordable Care Act of 2010.
The Patient Protection and Affordable Care Act includes several provisions that promise to stem the rapidly rising tide of uninsured young adults, one of the largest uninsured segments of the population. These include the ability to enroll in a parent's health plan up to age 26, beginning in September 2010; significant expansion in eligibility for Medicaid, beginning in 2014; and the creation of health insurance exchanges with subsidized private insurance for people with low and moderate incomes, also beginning in 2014. Of the 14.8 million uninsured young adults, up to 12.1 million may gain subsidized insurance once all the law's provisions go into effect in 2014: 7.2 million may gain coverage under Medicaid and 4.9 million may gain subsidized private coverage through the insurance exchanges. In addition, about 1 million uninsured young adults are expected to join their parents' policies over the next three years.


Dentin matrix protein 1 gene cis-regulation: use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo.
Dentin matrix protein 1 (DMP1) is highly expressed in osteocytes and is mechanically responsive. To study osteocyte-specific and mechanically regulated DMP1 gene expression, the transcriptional activity of three cis-regulatory regions was first examined in an osteoblast differentiation model in vitro using a green fluorescent protein (GFP) reporter. Expression of the -9624 to +1996 bp (10 kb) and -7892 to +4439 bp (8 kb) DMP1 cis-regulatory regions dramatically increased in areas of mineralized matrix, in dendritic, osteocyte-like cells. Mineralizing cultures expressing the 8-kb construct show dramatic GFP increases in response to loading in cells with a dendritic morphology. Transgenic mice expressing the 8-kb DMP1-GFP and -2433 to +4439 bp (2.5 kb) DMP1-LacZ were generated. Osteocyte-specific expression was found with the 8 kb but not with the 2.5 kb in postnatal animals. However, the 2.5 kb could support expression in rapidly forming osteoblasts and pre-osteocytes in the embryo. Primary calvarial osteoblast cultures demonstrated that the 2.5 kb supports weak expression in a subset of osteoblasts and pre-osteocytes, but not in mature osteocytes. However, the 8 kb supports robust expression in primary bone marrow cultures. Therefore the region -7892 to -2433 bp, termed a 5.5-kb "Osteocyte Enhancer Module," appears to be required for osteocyte specificity. Ulnae of mice with the 8-kb DMP1-GFP were subjected to mechanical loading where GFP expression increased selectively and locally in osteocytes, distal to the mid-shaft and near the surface of the bone. Thus, the 8-kb region of the DMP1 gene is a target for mechanotransduction in osteocytes, and its cis-regulatory activity may be correlated to local strain in bone.


Emerging Roles for MicroRNAs in Diabetic Microvascular Disease: Novel Targets for Therapy.
Chronic, low-grade systemic inflammation and impaired microvascular function are critical hallmarks in the development of insulin resistance. Accordingly, insulin resistance is a major risk factor for type 2 diabetes and cardiovascular disease. Accumulating studies demonstrate that restoration of impaired function of the diabetic macro- and microvasculature may ameliorate a range of cardiovascular disease states and diabetes-associated complications. In this review, we focus on the emerging role of microRNAs (miRNAs), noncoding RNAs that fine-tune target gene expression and signaling pathways, in insulin-responsive tissues and cell types important for maintaining optimal vascular homeostasis and preventing the sequelae of diabetes-induced end organ injury. We highlight current pathophysiological paradigms of miRNAs and their targets involved in regulating the diabetic microvasculature in a range of diabetes-associated complications such as retinopathy, nephropathy, wound healing, and myocardial injury. We provide an update of the potential use of circulating miRNAs diagnostically in type I or type II diabetes. Finally, we discuss emerging delivery platforms for manipulating miRNA expression or function as the next frontier in therapeutic intervention to improve diabetes-associated microvascular dysfunction and its attendant clinical consequences.


Hypolipidemic effect of Rukshana Upakrama in the management of dyslipidemia - A case study.
An elevated total or low-density lipoprotein (LDL) cholesterol levels or low levels of high-density lipoprotein (HDL) cholesterol is defined as dyslipidemia which is an important risk factor for coronary artery diseases (CADs) and strokes. It is estimated that abnormal cholesterol levels in the blood lead to cause about 18% of cardio vascular diseases (CVDs) and 56% of the ischemic heart diseases at global level. In comparison with others, South Asians have the highest incidence rate of CADs. Acharya Charaka has mentioned Rukshana Upakrama (a type of management which induces dryness) as one amongst the Shad-Upakrama (six principles of management) and can be considered as the best for the reduction of increased Kapha Dosha, Meda Dhatu (adipose tissue) and their related morbid factors. A 42-year-old female patient visited the OPD of the department of Basic Principles, IPGT and RA, Jamnagar, Gujarat, with the complaints of weight gain, heaviness in the body and lethargy. After investigation, she was diagnosed as a case of dyslipidemia due to abnormality in cholesterol levels. She was treated with Takra-Siddha Yavagu (gruel prepared with buttermilk) and Udvartana (dry powder massage) for 30 days. After completion of treatment, hematological investigations revealed that her serum cholesterol level was decreased to 147 mg/dl from 223 mg/dl and serum LDL was reduced to 91 mg/dl from 153 mg/dl. Her weight also reduced to 82 kgs from 88 kgs. Hence, it can be concluded that Rukshana Upakrama in the form of Takra-Siddha Yavagu and Udvartana is effective in the management of dyslipidemia for normalization of lipid in the blood as it possesses Shoshana (absorption) and Kapha-Medohara (alleviation of vitiated Kapha and Meda) properties.


A second wave of COVID-19 in Cook County: What lessons can be applied?
During the ongoing public health crisis, many agencies are reporting COVID-19 health outcome information based on the overall population. This practice can lead to misleading results and underestimation of high risk areas. To gain a better understanding of spatial and temporal distribution of COVID-19 deaths; the long term care facility (LTCF) and household population (HP) deaths must be used. This approach allows us to better discern high risk areas and provides policy makers with reliable information for community engagement and mitigation strategies. By focusing on high-risk LTCFs and residential areas, protective measures can be implemented to minimize COVID-19 spread and subsequent mortality. These areas should be a high priority target when COVID-19 vaccines become available.


Panning phage antibody libraries on cells: isolation of human Fab fragments against ovarian carcinoma using guided selection.
The display of repertoires of human antibody (Ab) fragments on filamentous phages and selection by binding of the phage to antigen (Ag) have provided a ready means of deriving human Ab against purified Ag. However, it has been more difficult to obtain phage Ab against an individual Ag of a complex mixture, such as cell surface Ag. Using the technique of "guided selection," we generated human Ab against the high-affinity folate-binding protein (FBP), a cell surface Ag that is overexpressed in many human ovarian carcinomas. The guiding Ab template was provided by the light chain of mouse monoclonal Ab Mov19 (K[aff], 10[8] M[-1]) directed against FBP; this was paired with repertoires of human heavy chains displayed on phages, and the hybrid Ab fragments were selected by binding to an ovarian carcinoma cell line (OVCAR3). The selected human heavy chains were then paired with repertoires of human light chains. Further panning led to the isolation of a human Fab fragment, C4, with a binding affinity of 0.2 x 10(8) M(-1). This was highly specific for FBP, as demonstrated by ELISA and flow cytometry data and by immunoprecipitation of the relevant molecule from the cell surface of ovarian carcinoma cells. Moreover, C4 targeted the same or a closely related epitope of the Ag, as did the template rodent monoclonal Ab Mov19. These results suggest the usefulness of guided selection as a simple means to deriving human Ab against cell surface Ag for which a rodent Ab is available.


High risk of suicide among high-school learners in uMgungundlovu District, KwaZulu-Natal Province, South Africa.
Worldwide, suicide affects the most vulnerable populations, including adolescents and young adults. It is estimated that suicide will contribute more than 2% to the global burden of disease by 2020. Information about the prevalence of suicidal behaviour and the sociodemographic characteristics and risk factors associated with suicide in the South African (SA) rural context is important for local and national policy and contributes to global understanding of the phenomenon. To investigate key demographic factors and behaviours associated with planning and attempting suicide among high-school learners. In a cross-sectional study, we used stratified random sampling to select 16 schools in uMgungundlovu District, KwaZulu-Natal Province, SA. All grade 10 learners (N=1 759) at these schools completed a self-administered questionnaire (Centers for Disease Control and Prevention (CDC) Youth Risk Behavior Surveillance System). Data analysis was carried out with Stata 13 statistical software using generalised estimating equations. In total, 222 learners (12.6% of the 1 759) had made plans to attempt suicide during the previous 12 months, 261 (14.8%) had actually attempted suicide, and 218 attempts had resulted in the learner being treated by a doctor or nurse (12.4%). The risk of planning suicide increased with age. For male learners, being threatened with a weapon on school property (adjusted odds ratio (AOR) 3.7, 95% confidence interval (CI) 1.9 - 7.1; p<0.01) or bullied through Facebook or WhatsApp (AOR 3.1, 95% CI 1.5 - 7.1; p<0.01) significantly increased the likelihood of making a suicide attempt that resulted in treatment by a doctor or nurse. For female learners, engaging in risk behaviours increased this likelihood, risk factors including being physically hurt by someone they were dating (1 - 3 times AOR 3.3, 95% CI 1.9 - 5.7; p<0.01, ≥4 times AOR 10.0, 95% CI 2.5 - 40.2 (p<0.01) and number of drinks consumed in the past month (AOR 2.0, 95% CI 1.4 - 3.0; p<0.01). The prevalence of suicide attempts among these SA learners was high and was influenced by multiple factors. Routine surveillance systems are urgently required to develop context-based interventions for male and female learners at uMgungundlovu District rural high schools.


Salmonella acid shock proteins are required for the adaptive acid tolerance response.
Salmonella typhimurium, as well as other enteric bacteria, experiences significant fluctuations in H+ ion concentrations during growth in diverse ecological niches. In fact, some pH conditions which should kill cells rapidly, such as stomach acidity, are nevertheless tolerated. The complete mechanism for this tolerance is unknown. However, I have recently demonstrated that S. typhimurium has the ability to survive extreme low pH (pH 3.0 to 4.0) if first adapted to mild pH (pH 5.5 to 6.0). This phenomenon has been referred to as the acidification tolerance response (ATR). The exposure to mild acid is referred to as preshock, and the proteins involved are called preshock ATR proteins. A second type of encounter with acid, called acid shock, involves shifting cells directly from alkaline conditions (pH 7.7) to acid conditions (pH 4.5 or below). During acid shock, the organism immediately ceases reproduction and dramatically changes the expression of at least 52 proteins. All but four are distinct from the preshock ATR proteins. Surprisingly, acid shock alone did not afford significant protection against strong acid challenge in minimal medium. Furthermore, inhibiting protein synthesis prior to acid shock revealed that the acid shock proteins do not appear to contribute to acid survival in minimal medium even at pH 4.3. Constitutive cellular pH homeostatic mechanisms seem sufficient to protect cells at this pH. The data suggest that the induction of acid shock and preshock ATR proteins are separate processes requiring separate signals. However, for S. typhimurium to survive extreme acid conditions, it must induce both the preshock and acid shock systems. Preventing the expression of one or the other eliminates acid tolerance. I propose a two-stage process that allows S. typhimurium to phase in acid tolerance as the environmental pH becomes progressively more acidic.


Sexual and physical abuse history in subjects with temporomandibular disorders: relationship to clinical variables, pain sensitivity, and psychologic factors.
Recent evidence suggests that a past history of physical and/or sexual abuse is more frequently reported among chronic pain populations; however, the prevalence of reported abuse has not been examined in patients with chronic orofacial pain caused by temporomandibular disorders (TMD). This study compares reported physical/sexual abuse among female TMD subjects recruited from the general population with that of age-matched female control subjects. The association of reported abuse with clinical pain, experimental pain responses, and psychologic variables was examined in the TMD group. Results indicated that a slightly but not statistically greater percentage of TMD subjects (44.8%) reported a history of sexual or physical abuse compared to control subjects (33.3%). Reported abuse among TMD subjects was not related to clinical pain or psychologic variables. Regarding experimental pain responses, TMD subjects reporting a history of abuse exhibited longer ischemic pain tolerances compared to those not reporting abuse; however, the groups did not differ on other experimental pain measures. Results indicate that the reported prevalence of physical/sexual abuse is similar among TMD subjects compared to other chronic pain populations; however, the relationship of abuse to clinical and psychosocial variables remains unclear.


Hsp70 accumulation in chondrocytic cells exposed to high continuous hydrostatic pressure coincides with mRNA stabilization rather than transcriptional activation.
In response to various stress stimuli, heat shock genes are induced to express heat shock proteins (Hsps). Previous studies have revealed that expression of heat shock genes is regulated both at transcriptional and posttranscriptional level, and the rapid transcriptional induction of heat shock genes involves activation of the specific transcription factor, heat shock factor 1 (HSF1). Furthermore, the transcriptional induction can vary in intensity and kinetics in a signal- and cell-type-dependent manner. In this study, we demonstrate that mechanical loading in the form of hydrostatic pressure increases heat shock gene expression in human chondrocyte-like cells. The response to continuous high hydrostatic pressure was characterized by elevated mRNA and protein levels of Hsp70, without activation of HSF1 and transcriptional induction of hsp70 gene. The increased expression of Hsp70 was mediated through stabilization of hsp70 mRNA molecules. Interestingly, in contrast to static pressurization, cyclic hydrostatic loading did not result in the induction of heat shock genes. Our findings show that hsp70 gene expression is regulated posttranscriptionally without transcriptional induction in chondrocyte-like cells upon exposure to high continuous hydrostatic pressure. We suggest that the posttranscriptional regulation in the form of hsp70 mRNA stabilization provides an additional mode of heat shock gene regulation that is likely to be of significant importance in certain forms of stress.


Latest developments in our understanding of the pathogenesis of mesothelioma and the design of targeted therapies.
Malignant mesothelioma is an aggressive cancer whose pathogenesis is causally linked to occupational exposure to asbestos. Familial clusters of mesotheliomas have been observed in settings of genetic predisposition. Mesothelioma incidence is anticipated to increase worldwide in the next two decades. Novel treatments are needed, as current treatment modalities may improve the quality of life, but have shown modest effects in improving overall survival. Increasing knowledge on the molecular characteristics of mesothelioma has led to the development of novel potential therapeutic strategies, including: molecular targeted approaches, that is the inhibition of vascular endothelial growth factor with bevacizumab; immunotherapy with chimeric monoclonal antibody, immunotoxin, antibody drug conjugate, vaccine and viruses; inhibition of asbestos-induced inflammation, that is aspirin inhibition of HMGB1 activity may decrease or delay mesothelioma onset and/or growth. We elaborate on the rationale behind new therapeutic strategies, and summarize available preclinical and clinical results, as well as efforts still ongoing.


A microfluidic fluorescence measurement system using an astigmatic diffractive microlens array.
We demonstrate an opto-fluidic detection system based on an array of astigmatic diffractive microlenses integrated into a microfluidic flow focus device. Each astigmatic microlens produces a line excitation across the channel and collects fluorescence emission from the linear detection regions. The linear excitation spot results in uniform excitation across the channel and high time resolution in the direction of the flow. Collected fluorescence from each integrated microlens is relayed to a sub-region on a fast CMOS camera. By analyzing the signal from individual microlenses, we demonstrate counting and resolution of 500 nm and 1.1 μm beads at rates of up to 8,300 per second at multiple locations. In addition, a cross-correlation analysis of the signals from different microlenses yields the velocity dispersion of beads traveling through the channel at peak speeds as high as 560 mm/s. Arrays of specifically designed diffractive optics promise to increase the resolution and functionality of opto-fluidic analysis such as flow cytometry and fluorescence cross-correlation spectroscopy.


First report on Encephalitozoon intestinalis infection of swine in Europe.
Encephalitozoon intestinalis infection of sows is reported from a pig farm in Slovakia. Spores were detected by direct microscopic visualisation in the faeces of 25 out of 27 sows (92.6%). This finding was also supported serologically by the presence of specific anti-E. intestinalis antibodies and by a species-specific polymerase chain reaction (PCR). This is the first report on E. intestinalis infection of swine in Europe.


Daratumumab, lenalidomide, bortezomib, and dexamethasone for transplant-eligible newly diagnosed multiple myeloma: the GRIFFIN trial.
Lenalidomide, bortezomib, and dexamethasone (RVd) followed by autologous stem cell transplantation (ASCT) is standard frontline therapy for transplant-eligible patients with newly diagnosed multiple myeloma (NDMM). The addition of daratumumab (D) to RVd (D-RVd) in transplant-eligible NDMM patients was evaluated. Patients (N = 207) were randomized 1:1 to D-RVd or RVd induction (4 cycles), ASCT, D-RVd or RVd consolidation (2 cycles), and lenalidomide or lenalidomide plus D maintenance (26 cycles). The primary end point, stringent complete response (sCR) rate by the end of post-ASCT consolidation, favored D-RVd vs RVd (42.4% vs 32.0%; odds ratio, 1.57; 95% confidence interval, 0.87-2.82; 1-sided P = .068) and met the prespecified 1-sided α of 0.10. With longer follow-up (median, 22.1 months), responses deepened; sCR rates improved for D-RVd vs RVd (62.6% vs 45.4%; P = .0177), as did minimal residual disease (MRD) negativity (10-5 threshold) rates in the intent-to-treat population (51.0% vs 20.4%; P < .0001). Four patients (3.8%) in the D-RVd group and 7 patients (6.8%) in the RVd group progressed; respective 24-month progression-free survival rates were 95.8% and 89.8%. Grade 3/4 hematologic adverse events were more common with D-RVd. More infections occurred with D-RVd, but grade 3/4 infection rates were similar. Median CD34+ cell yield was 8.2 × 106/kg for D-RVd and 9.4 × 106/kg for RVd, although plerixafor use was more common with D-RVd. Median times to neutrophil and platelet engraftment were comparable. Daratumumab with RVd induction and consolidation improved depth of response in patients with transplant-eligible NDMM, with no new safety concerns. This trial was registered at www.clinicaltrials.gov as #NCT02874742.


Congenital pituitary hypoplasia model demonstrates hypothalamic OTX2 regulation of pituitary progenitor cells.
Pituitary develops from oral ectoderm in contact with adjacent ventral hypothalamus. Impairment in this process results in congenital pituitary hypoplasia (CPH); however, there have been no human disease models for CPH thus far, prohibiting the elucidation of the underlying mechanisms. In this study, we established a disease model of CPH using patient-derived induced pluripotent stem cells (iPSCs) and 3D organoid technique, in which oral ectoderm and hypothalamus develop simultaneously. Interestingly, patient iPSCs with a heterozygous mutation in the orthodenticle homeobox 2 (OTX2) gene showed increased apoptosis in the pituitary progenitor cells, and the differentiation into pituitary hormone-producing cells was severely impaired. As an underlying mechanism, OTX2 in hypothalamus, not in oral ectoderm, was essential for progenitor cell maintenance by regulating LHX3 expression in oral ectoderm via FGF10 expression in the hypothalamus. Convincingly, the phenotype was reversed by the correction of the mutation, and the haploinsufficiency of OTX2 in control iPSCs revealed a similar phenotype, demonstrating that this mutation was responsible. Thus, we established an iPSC-based congenital pituitary disease model, which recapitulated interaction between hypothalamus and oral ectoderm and demonstrated the essential role of hypothalamic OTX2.


Expression patterns of bovine CD1 in vivo and assessment of the specificities of the anti-bovine CD1 antibodies.
Research addressing the in vivo effects of T cell activation by lipids, glycolipids, and lipopeptides is hampered by the absence of a suitable animal model. Mice and rats do not express CD1a, CD1b, and CD1c molecules that present pathogen-derived lipid antigens in humans. In cattle, two CD1A and three CD1B genes are transcribed. The proteins encoded by these genes differ in their antigen binding domains and in their cytoplasmic tails, suggesting that they may traffic differently in the cell and thus have access to different antigens. In the current study, we describe the genomic organization of the bovine CD1 locus and transcription of bovine CD1 genes in freshly isolated dendritic cells and B cells from different tissues. After determining the specificity of previously only partly characterized anti-CD1 antibodies by testing recombinant single chain bovine CD1 proteins and CD1-transfected cells, we were able to determine cell surface protein expression on freshly isolated cells. Our study suggests that CD1b1 and CD1b3 are more broadly expressed than CD1b5, and CD1a2 is more broadly expressed than CD1a1. Pseudoafferent lymph dendritic cells express CD1B genes, but no transcription is detected in lymph nodes. Even though B cells transcribe CD1B genes, there is no evidence of protein expression at the cell surface. Thus, patterns of CD1 protein expression are largely conserved among species.


The Effects of Low Birth Weight on School Performance and Behavioral Outcomes of Elementary School Children in Oman.
Our study aimed to examine the effects of low birth weight (LBW) on the school performance and behavior of elementary school children in Oman. . Data were gathered through a cross-sectional survey of nine elementary schools from the Muscat and A'Dhahirah regions. The study utilized a unique database created by linking information from the children's health cards and current academic and behavioral performance records. Information on children's performance in various areas such as language, mathematics, science, information technology, sports, and behavior were obtained from the school registers. Birth weight (BW) and selected sociodemographic data were obtained from the copy of their health cards kept by each school. A total of 542 elementary school children aged 7-11 years, who had completed grades 2-4, were surveyed. . Data from the school register revealed a very high rate (17.7%) of LBW and, overall, 12% of the children exhibited below average performance on selected outcome measures. The below average school performance varied from 5-17% across the six selected areas of school performance. The highest rate of below average performance was observed in science (17%), followed by arithmetic and language (16% each). BW showed significant differential effects on school performance and behavioral outcomes, which remained significant after controlling for the effect of potential confounders. It was found that LBW children were 2-6 times more likely to have poorer school performance in all areas than their normal BW peers. . Early intervention programs or special care for LBW children in school could be an effective means of improving educational outcomes and the behavior of these children. Attempts should be made to reduce or prevent poor pregnancy outcomes, which, in turn, would reduce the cost of the health, education, and social services systems.


The interacting effects of food, spring temperature, and global climate cycles on population dynamics of a migratory songbird.
Although long-distance migratory songbirds are widely believed to be at risk from warming temperature trends, species capable of attempting more than one brood in a breeding season could benefit from extended breeding seasons in warmer springs. To evaluate local and global factors affecting population dynamics of the black-throated blue warbler (Setophaga caerulescens), a double-brooded long-distance migrant, we used Pradel models to analyze 25 years of mark-recapture data collected in New Hampshire, USA. We assessed the effects of spring temperature (local weather) and the El Niño Southern Oscillation index (a global climate cycle), as well as predator abundance, insect biomass, and local conspecific density on population growth in the subsequent year. Local and global climatic conditions affected warbler populations in different ways. We found that warbler population growth was lower following El Niño years (which have been linked to poor survival in the wintering grounds and low fledging weights in the breeding grounds) than La Niña years. At a local scale, populations increased following years with warm springs and abundant late-season food, but were unaffected by spring temperature following years when food was scarce. These results indicate that the warming temperature trends might have a positive effect on recruitment and population growth of black-throated blue warblers if food abundance is sustained in breeding areas. In contrast, potential intensification of future El Niño events could negatively impact vital rates and populations of this species.


Toward outcomes-based plastic surgery training: a needs assessment of recent graduates.
The Accreditation Council for Graduate Medical Education Outcomes Project mandates data-driven improvements to training programs. Few outcomes measures exist in plastic surgery. The purpose of this study was to determine how well we presently train plastic surgery residents within the context of the council's competencies and how well those competencies are practiced after residency. Plastic surgery graduates from 2000 to 2005 were surveyed using a new questionnaire organized around the accreditation council's competencies. Scores from individual item responses were averaged to create a composite score reflecting the quality of training and practice of each competency. Composite scores were then compared using nonparametric analyses. Data regarding currently used assessment tools were also measured. The response rate was 21 percent. The respondent sample was representative of current plastic surgeons. Overall, the competencies were considered important, with increasing awareness in more recent years. Practice-based learning and improvement appeared to be the best taught, and systems-based practice appeared to be the most poorly taught. There were differences between independent and integrated formats but they did not support one over the other. Women did not rate their training as highly as men did in some areas, and open comments suggested there were significant issues at some programs. Recent graduates view plastic surgery training as excellent. The competency domain in greatest need of attention is systems-based practice. We must broaden our use of assessment instruments for more valid results and investigate gender differences in our training programs. These results describe the outcomes of current plastic surgery training and serve as a needs assessment for improvement and as a baseline for future comparison.


Geoarchaeota: a new candidate phylum in the Archaea from high-temperature acidic iron mats in Yellowstone National Park.
Geothermal systems in Yellowstone National Park (YNP) provide an outstanding opportunity to understand the origin and evolution of metabolic processes necessary for life in extreme environments including low pH, high temperature, low oxygen and elevated concentrations of reduced iron. Previous phylogenetic studies of acidic ferric iron mats from YNP have revealed considerable diversity of uncultivated and undescribed archaea. The goal of this study was to obtain replicate de novo genome assemblies for a dominant archaeal population inhabiting acidic iron-oxide mats in YNP. Detailed analysis of conserved ribosomal and informational processing genes indicates that the replicate assemblies represent a new candidate phylum within the domain Archaea referred to here as 'Geoarchaeota' or 'novel archaeal group 1 (NAG1)'. The NAG1 organisms contain pathways necessary for the catabolism of peptides and complex carbohydrates as well as a bacterial-like Form I carbon monoxide dehydrogenase complex likely used for energy conservation. Moreover, this novel population contains genes involved in the metabolism of oxygen including a Type A heme copper oxidase, a bd-type terminal oxidase and a putative oxygen-sensing protoglobin. NAG1 has a variety of unique bacterial-like cofactor biosynthesis and transport genes and a Type3-like CRISPR system. Discovery of NAG1 is critical to our understanding of microbial community structure and function in extant thermophilic iron-oxide mats of YNP, and will provide insight regarding the evolution of Archaea in early Earth environments that may have important analogs active in YNP today.


Characterization of 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidases from Borrelia burgdorferi: Antibiotic targets for Lyme disease.
Borrelia burgdorferi causes Lyme disease, the most common tick-borne illness in the United States. The Center for Disease Control and Prevention estimates that the occurrence of Lyme disease in the U.S. has now reached approximately 300,000 cases annually. Early stage Borrelia burgdorferi infections are generally treatable with oral antibiotics, but late stage disease is more difficult to treat and more likely to lead to post-treatment Lyme disease syndrome. Here we examine three unique 5'-methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidases (MTNs or MTANs, EC 184.108.40.206) responsible for salvage of adenine and methionine in B. burgdorferi and explore their potential as antibiotic targets to treat Lyme disease. Recombinant Borrelia MTNs were expressed and purified from E. coli. The enzymes were extensively characterized for activity, specificity, and inhibition using a UV spectrophotometric assay. In vitro antibiotic activities of MTN inhibitors were assessed using a bioluminescent BacTiter-Glo™ assay. The three Borrelia MTNs showed unique activities against the native substrates MTA, SAH, and 5'-deoxyadenosine. Analysis of substrate analogs revealed that specific activity rapidly dropped as the length of the 5'-alkylthio substitution increased. Non-hydrolysable nucleoside transition state analogs demonstrated sub-nanomolar enzyme inhibition constants. Lastly, two late stage transition state analogs exerted in vitro IC50 values of 0.3-0.4 μg/mL against cultured B. burgdorferi cells. B. burgdorferi is unusual in that it expresses three distinct MTNs (cytoplasmic, membrane bound, and secreted) that are effectively inactivated by nucleoside analogs. The Borrelia MTNs appear to be promising targets for developing new antibiotics to treat Lyme disease.


Factor's affecting safe emergency evacuation of subways in Iran: findings of a qualitative study.
Development of construction and operation of subway rail transit system in populated cities of middle- and high-income countries along with the increasing population of its users, have exacerbated the safety problems of the users against incidents and emergencies in subway stations. Although subway emergency evacuation is considered by the governments and subway network organizations as a critical task related to passengers' safety at the time of emergency, the risk of mass evacuation in station is undeniable. The main objective of this study is to identify factors affecting safe evacuation of the population from subway station and to propose the strategies for addressing them based on experiences or opinions of stakeholders in Tehran Subways, Iran. This is a qualitative study that was conducted between January 2017 and December 2018, in which a semi-structured interview was conducted for 17 participants among the senior managers, executive managers, subway station operations staff, and subway passengers in Tehran subway stations, in the Capital of Iran. In order to analyze the data of this qualitative study, the Graneheim and Lundman method was used and manifested content analysis approach was employed. Based on the findings of this study, the factors affecting safe evacuation of the population from subways station were identified in four main categories covering passengers, organization, communications, and environment. Then the main categories of "culture, interaction and cooperation of passengers", "correct and timely decision-making", "notification", and "location of emergency evacuation" were emerged as safe emergency evacuation challenges and the most important findings of this study; and strategies were proposed to improve the safety of passengers at emergency evacuation of subway stations. In the present study, the lack of safe approach to improving passengers' safety in the development plan of Tehran subway rail transit network is a major concern for managers and operations staff. Therefore, changing the attitude of policy makers from focusing on the quantitative development of passenger transportation services to improving safety and then the quality of passengers' trip is taken into account as an urgent need to improve the safety of subway passengers.