Elevated gamma band power in humans receiving naloxone suggests dorsal periaqueductal and periventricular gray deep brain stimulation produced analgesia is opioid mediated.
The midbrain periaqueductal and periventricular gray (PAVG) region is important for pain and autonomic modulation. We have previously described changes in blood pressure dependent upon dorsal or ventral electrode positioning with PAVG deep brain stimulation (DBS), yet controversy exists about whether DBS acts via endogenous opioid release. We combined local field potential (LFP) recording from PAVG DBS electrodes in humans with naloxone and saline infusions to determine whether dorsal and ventral PAVG DBS act through opioidergic or other mechanisms. Four awake human subjects were investigated. DBS were implanted contralateral to the painful body part. Electrode contact positions were mapped using MRI and brain atlas information. Naloxone then saline were randomly administered to the blinded subjects and pain rated using a numeric pain rating scale at 30s intervals for 3 min. Two subjects received dorsal DBS electrodes and two had ventral placements. Significantly elevated gamma frequency band (30-90 Hz) power concomitant with pain exacerbation was found with naloxone versus both saline and rest in dorsal not ventral PAVG LFPs (p<0.005). Significantly elevated delta frequency band (0-4 Hz) power (p=0.001) was seen in one ventral PAVG subject with both naloxone and saline infusions. Dorsal PAVG DBS may reduce pain by augmenting endogenous opioid release. Elevated gamma oscillations enhance awareness of worsening pain with opioid blockade. Ventral PAVG DBS may act by separate possibly autonomic mechanisms. Increased delta oscillations indicate a survival rhythm involved in the initiation of passive coping responses to homeostatic changes.


Canine sterile nodular panniculitis: a retrospective study of 14 cases.
Sterile nodular panniculitis (SNP) is an uncommon inflammatory condition of subcutaneous fat that can be idiopathic, but has also been associated with underlying conditions such as pancreatic disease or systemic lupus erythematosus (SLE). The pathogenesis and clinical course of the condition are not well understood. To retrospectively review cases of SNP associated with systemic signs, concurrent disease, or both and characterize the clinical, laboratory, imaging, and histopathologic findings, treatment, and response to treatment. Fourteen dogs with histologically confirmed SNP diagnosed between 1996 and 2008. Retrospective study. Skin lesions were ulcerated or draining nodules in 9 dogs and nonulcerative subcutaneous nodules in 5. Most dogs had systemic signs, such as fever, inappetence, lethargy, and multiple lesions. Common clinicopathologic findings included neutrophilia with or without left shift, increased alkaline phosphatase activity, mild hypoglycemia, hypoalbuminemia, and proteinuria. Concurrent diseases included pancreatic disease, SLE, rheumatoid arthritis, polyarthritis, lymphoplasmacytic colitis, and hepatic disease. Dogs responded to immunosuppressive doses of corticosteroids when administered. Prognosis for recovery was related to the underlying disease process. SNP is not a single disease. Rather, it is a cutaneous marker of systemic disease in many cases. After thorough evaluation for concurrent disease and infectious causes, immunosuppressive treatment is often effective.


Podometrics as a Potential Clinical Tool for Glomerular Disease Management.
Chronic kidney disease culminating in end-stage kidney disease is a major public health problem costing in excess of $40 billion per year with high morbidity and mortality. Current tools for glomerular disease monitoring lack precision and contribute to poor outcome. The podocyte depletion hypothesis describes the major mechanisms underlying the progression of glomerular diseases, which are responsible for more than 80% of cases of end-stage kidney disease. The question arises of whether this new knowledge can be used to improve outcomes and reduce costs. Podocytes have unique characteristics that make them an attractive monitoring tool. Methodologies for estimating podocyte number, size, density, glomerular volume and other parameters in routine kidney biopsies, and the rate of podocyte detachment from glomeruli into urine (podometrics) now have been developed and validated. They potentially fill important gaps in the glomerular disease monitoring toolbox. The application of these tools to glomerular disease groups shows good correlation with outcome, although data validating their use for individual decision making is not yet available. Given the urgency of the clinical problem, we argue that the time has come to focus on testing these tools for application to individualized clinical decision making toward more effective progression prevention.


Can tumor necrosis factor inhibitors be safely used in pregnancy?
We review available safety data for use of currently approved tumor necrosis factor (TNF) inhibitors during pregnancy and lactation and suggest guidelines for use of these agents among women of reproductive age. Method. Although regulatory agencies encourage the inclusion of pregnant women and those of child-bearing age in randomized controlled trials, pregnant and lactating women have universally been excluded from studies because of unknown or potential risks to the fetus. Thus, strong evidence-based treatment recommendations during pregnancy are usually lacking and safety information is derived from voluntary reports of adverse events during postmarketing surveillance or via uncontrolled, observational studies, reviewed here. Uncommon adverse pregnancy outcomes observed with TNF inhibitor therapy appear to approximate those seen in women not receiving such therapy and may include premature birth, miscarriage, low birthweight, hypertension, and preeclampsia. There are rare reports of fetal malformations or congenital anomalies in patients exposed to TNF inhibitors during conception or pregnancy. However, the incidence of these events appears to be far below the 3% rate of congenital anomalies in the general population. If the activity or disease severity precludes the cessation of a TNF inhibitor and/or DMARD, uncontrolled observations suggest that conception and early pregnancy are not adversely affected by use of TNF inhibitors. Nearly 70% of pregnant patients can discontinue their TNF inhibitor early in the pregnancy (or with determination of pregnancy) without augmenting maternal or fetal risks.


Detecting Large Chromosomal Modifications Using Short Read Data From Genotyping-by-Sequencing.
Markers linked to agronomic traits are of the prerequisite for molecular breeding. Genotyping-by-sequencing (GBS) data enables to detect small polymorphisms including single nucleotide polymorphisms (SNPs) and short insertions or deletions (InDels) that can be used, for instance, for marker-assisted selection, population genetics, and genome-wide association studies (GWAS). Here, we aim at detecting large chromosomal modifications in barley and wheat based on GBS data. These modifications could be duplications, deletions, substitutions including introgressions as well as alterations of DNA methylation. We demonstrate that GBS coverage analysis is capable to detect Hordeum vulgare/Hordeum bulbosum introgression lines. Furthermore, we identify large chromosomal modifications in barley and wheat collections. Hence, large chromosomal modifications, including introgressions and copy number variations (CNV), can be detected easily and can be used as markers in research and breeding without additional wet-lab experiments.


Racial stereotyping of indigenous people in the Canadian media: A comparative analysis of two water pollution incidents.
This article examines the discourse surrounding issues affecting Indigenous peoples within the Canadian mainstream media. We compare the coverage of two cases of water poisonings-one in a primarily-white town and the other in an Indigenous community-in 282 newspaper articles from the Toronto Star, the Globe and Mail, the National Post, and Windspeaker. We show that the dominant coverage of these two very similar cases diverged significantly. The Indigenous workers in charge of the water supply were regarded as incompetent and incapable to fill their position while the entire community was described as drunk, lazy, helpless, and perpetually dependent on government aide. By contrast, white workers were seen as relatable and in command of their erroneous actions, while the residents of the town were described simply as the victims of an unfortunate tragedy. Such reporting fails to contextualize the events or point out the injustices of Canadian colonialism, thus contributing to the perpetuation of these injustices.


Constitutive reduction in the checkpoint inhibitor, CTLA-4, does not accelerate SLE in NZM 2328 mice.
Treatment with immune checkpoint inhibitors (ICIs) in oncology patients is increasing. Although ICIs trigger rheumatic immune-related adverse events, development of SLE features has been rare. Whether long-term treatment with ICIs would promote SLE features remains unknown. To begin to address this, we generated SLE-prone NZM 2328 mice with lifelong reduction in CTLA-4 expression. Since CTLA-4-deficient (Ctla4- /-) NZM mice developed a lethal lymphoproliferative disorder by 3-6 weeks of age, development of SLE in these mice could not be studied. Ctla4 haploinsufficient NZM.Ctla4+ / - mice were assessed in parallel with littermate female NZM.Ctla4+ / + mice. Evaluations included CTLA-4 expression and lymphocyte profiles, assessed by fluorescence-activated cell sorting; serological profiles, assessed by ELISA; renal immunopathology, assessed by histology and immunofluorescence; and clinical courses, assessed by mortality. CTLA-4 expression was lower in NZM.Ctla4+ / - mice than in NZM.Ctla4+ / + mice. Spleen mononuclear cells, B cells, plasma cells, CD4+ cells, recently activated CD4+ cells and CD4+ T regulatory (Treg) cells were increased in NZM.Ctla4+ / - mice (p≤0.042). The serological profile, degree of renal immunopathology and mortality in NZM.Ctla4+ / - mice remained unaffected. Lifelong reduction in CTLA-4 expression in NZM mice neither accelerated nor aggravated SLE. Expansion in Treg cells may have played a protective role. Our observations raise the hope that long-term treatment of patients with SLE with an anti-CTLA-4 agent, should the need arise, would not adversely affect SLE disease activity.


A rapid and low-cost fabrication and integration scheme to render 3D microfluidic architectures for wearable biofluid sampling, manipulation, and sensing.
The large-scale deployment of wearable bioanalytical devices for general population longitudinal monitoring necessitates rapid and high throughput manufacturing-amenable fabrication schemes that render disposable, low-cost, and mechanically flexible microfluidic modules capable of performing a variety of bioanalytical operations within a compact footprint. The spatial constraints of previously reported wearable bioanalytical devices (with microfluidic operations confined to 2D), their lack of biofluid manipulation capability, and the complex and low-throughput nature of their fabrication process inherently limit the diversity and frequency of end-point assessments and prevent their deployment at large scale. Here, we devise a simple, scalable, and low-cost "CAD-to-3D Device" fabrication and integration scheme, which renders 3D and complex microfluidic architectures capable of performing biofluid sampling, manipulation, and sensing. The devised scheme is based on laser-cutting of tape-based substrates, which can be programmed at the software-level to rapidly define microfluidic features such as a biofluid collection interface, microchannels, and VIAs (vertical interconnect access), followed by the vertical assembly of pre-patterned layers to realize the final device. To inform the utility of our fabrication scheme, we demonstrated three representative devices to perform sweat collection (with visualizable secretion profile), sample filtration, and simultaneous biofluid actuation and sensing (using a sandwiched-interface). Our devised scheme can be adapted for the fabrication and manufacturing of current and future wearable bioanalytical devices, which in turn will catalyze the large-scale production and deployment of such devices for general population health monitoring.


Rationale and design of a randomized, double-blind, parallel-group study of terutroban 30 mg/day versus aspirin 100 mg/day in stroke patients: the prevention of cerebrovascular and cardiovascular events of ischemic origin with terutroban in patients with a history of ischemic stroke or transient ischemic attack (PERFORM) study.
Ischemic stroke is the leading cause of mortality worldwide and a major contributor to neurological disability and dementia. Terutroban is a specific TP receptor antagonist with antithrombotic, antivasoconstrictive, and antiatherosclerotic properties, which may be of interest for the secondary prevention of ischemic stroke. This article describes the rationale and design of the Prevention of cerebrovascular and cardiovascular Events of ischemic origin with teRutroban in patients with a history oF ischemic strOke or tRansient ischeMic Attack (PERFORM) Study, which aims to demonstrate the superiority of the efficacy of terutroban versus aspirin in secondary prevention of cerebrovascular and cardiovascular events. The PERFORM Study is a multicenter, randomized, double-blind, parallel-group study being carried out in 802 centers in 46 countries. The study population includes patients aged > or =55 years, having suffered an ischemic stroke (< or =3 months) or a transient ischemic attack (< or =8 days). Participants are randomly allocated to terutroban (30 mg/day) or aspirin (100 mg/day). The primary efficacy endpoint is a composite of ischemic stroke (fatal or nonfatal), myocardial infarction (fatal or nonfatal), or other vascular death (excluding hemorrhagic death of any origin). Safety is being evaluated by assessing hemorrhagic events. Follow-up is expected to last for 2-4 years. Assuming a relative risk reduction of 13%, the expected number of primary events is 2,340. To obtain statistical power of 90%, this requires inclusion of at least 18,000 patients in this event-driven trial. The first patient was randomized in February 2006. The PERFORM Study will explore the benefits and safety of terutroban in secondary cardiovascular prevention after a cerebral ischemic event.


Serumfree culture of the suspension cell line QB-Tn9-4s of the cabbage looper, Trichoplusia ni, is highly productive for virus replication and recombinant protein expression.
Serumfree cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serumfree Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 ×10(6) cells/mL in serumfree culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 10(6) cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 ×10(7) TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of β-galactosidase at six days postinfection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×10(4) IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serumfree culture.


Attosecond electron dynamics.
We describe the recent emergence of attosecond science, assessing the present state of the art and discussing several recent examples where attosecond electron dynamics has been studied in atomic and molecular systems. After introducing the generation and characterization of attosecond laser pulses, we describe the use of isolated attosecond pulses in a pump-probe experiment revealing the subcycle time dependence of a multiphoton ionization process and an experiment using the interference from a train of attosecond pulses to extract amplitude and phase information for electronic wave functions. We furthermore discuss experiments where ultrashort laser pulses with a reproducible waveform control electron dynamics in the D(2)(+) molecular ion on attosecond timescales. Attosecond science is coming of age and presently is reaching a level of maturity and sophistication that allows detailed investigations of the role of multielectron dynamics in physics and chemistry.


Evidence for production of single top quarks and first direct measurement of |Vtb|.
The D0 Collaboration presents first evidence for the production of single top quarks at the Fermilab Tevatron pp[over ] collider. Using a 0.9 fb(-1) dataset, we apply a multivariate analysis to separate signal from background and measure sigma(pp[over ]-->tb+X,tqb+X)=4.9+/-1.4 pb. The probability to measure a cross section at this value or higher in the absence of a signal is 0.035%, corresponding to a 3.4 standard deviation significance. We use the cross section measurement to directly determine the Cabibbo-Kobayashi-Maskawa matrix element that describes the Wtb coupling and find 0.68<|V(tb)|</=1 at 95% C.L. within the standard model.


Continuous Indexing of Fibrosis (CIF): improving the assessment and classification of MPN patients.
The grading of fibrosis in myeloproliferative neoplasms (MPN) is an important component of disease classification, prognostication and monitoring. However, current fibrosis grading systems are only semi-quantitative and fail to fully capture sample heterogeneity. To improve the quantitation of reticulin fibrosis, we developed a machine learning approach using bone marrow trephine (BMT) samples (n = 107) from patients diagnosed with MPN or a reactive marrow. The resulting Continuous Indexing of Fibrosis (CIF) enhances the detection and monitoring of fibrosis within BMTs, and aids MPN subtyping. When combined with megakaryocyte feature analysis, CIF discriminates between the frequently challenging differential diagnosis of essential thrombocythemia (ET) and pre-fibrotic myelofibrosis with high predictive accuracy [area under the curve = 0.94]. CIF also shows promise in the identification of MPN patients at risk of disease progression; analysis of samples from 35 patients diagnosed with ET and enrolled in the Primary Thrombocythemia-1 trial identified features predictive of post-ET myelofibrosis (area under the curve = 0.77). In addition to these clinical applications, automated analysis of fibrosis has clear potential to further refine disease classification boundaries and inform future studies of the micro-environmental factors driving disease initiation and progression in MPN and other stem cell disorders.


Microbial hydroxylation of 5-anilino-1,2,3,4-thiatriazole.
Two hundred eighty-five fungi, including 100 basidiomycetes and 35 yeasts, 75 actinomycetes, and 40 bacteria were screened for their ability to convert 5-anilino-1,2,3,4-thiatriazole (AT) to 5-(p-hydroxyanilino)-1,2,3,4-thiatriazole (p-HT). Eleven cultures were found that formed p-HT, which was isolated and whose structure was determined. Aspergillus tamarii NRRL 3280 formed 8.6 g of p-HT/liter from 10 g of AT/liter (78.9% conversion) in shaken flasks and 4.57 g of p-HT/liter from 6 g of AT/liter (69.8% conversion) in 30-liter fermentors. Washed cells of A. tamarii NRRL 3280 also carried out this conversion. 5-(o-hydroxyanilino)-1,2,3,4-thiatriazole (o-HT) was identified as a second product formed by Aspergillus terreus NRRL 1960.


Magnetic seizure therapy (MST)--a safer method for evoking seizure activity than current therapy with a confirmed antidepressant efficacy.
Since 1999, attempts have been made in the application of a new technique called magnetic seizure therapy (MST) or magnetic convulsion therapy (MCT) in the treatment of depressive disorder--as an alternative to electroconvulsive treatment. The technique of rapid rate transcranial magnetic stimulation (rTMS) is used to evoke intentional and repeated magnetoconvulsive seizures, though it requires the use of stimulation parameters practically inaccessible in commercially available rTMS magnetic stimulators. Magnetic convulsion therapy has been tested on monkeys as well as humans. A decisive majority of studies carried out both on animals and humans addressed the issue of safety of the MST method and confirmed that the side-effects (mostly of a cognitive nature) which occurred after magnetic seizures were weaker than those observed after electroconvulsive seizures. An analysis of available sources, however, does not confirm any proven antidepressant action of the MST technique. No experimental investigations have been carried out on animal models of depression. Clinical effectiveness had been confirmed in merely a few (perhaps three) patients with depression. The authors submit the results of the hitherto conducted studies on MST to critical analysis, particularly in the aspect of their antidepressant efficacy.


Microfiber Masses Recovered from Conventional Machine Washing of New or Aged Garments.
Synthetic textiles can shed numerous microfibers during conventional washing, but evaluating environmental consequences as well as source-control strategies requires understanding mass releases. Polyester apparel accounts for a large proportion of the polyester market, and synthetic jackets represent the broadest range in apparel construction, allowing for potential changes in manufacturing as a mitigation measure to reduce microfiber release during laundering. Here, detergent-free washing experiments were conducted and replicated in both front- and top-load conventional home machines for five new and mechanically aged jackets or sweaters: four from one name-brand clothing manufacturer (three majority polyester fleece, and one nylon shell with nonwoven polyester insulation) and one off-brand (100% polyester fleece). Wash water was filtered to recover two size fractions (>333 μm and between 20 and 333 μm); filters were then imaged, and microfiber masses were calculated. Across all treatments, the recovered microfiber mass per garment ranged from approximately 0 to 2 g, or exceeding 0.3% of the unwashed garment mass. Microfiber masses from top-load machines were approximately 7 times those from front-load machines; garments mechanically aged via a 24 h continuous wash had increased mass release under the same wash protocol as new garments. When published wastewater treatment plant influent characterization and microfiber removal studies are considered, washing synthetic jackets or sweaters as per this study would account for most microfibers entering the environment.


Effects of lactate on glucose metabolism of developing rat brain.
The effects of hyperlactatemia on cerebral glucose metabolism of normoglycemic 20-day-old rats were studied in animals breathing air or 20% CO2:21% O2:59% N2. Sodium lactate or sodium bicarbonate were given intraperitoneally, together with a mixture of [3H]deoxyglucose and [2-14C]glucose. Animals were sacrificed in a freeze-blowing apparatus at intervals of 2-15 min after injection. Blood lactate levels in the lactate-injected rats were 4-6 mM. Hyperlactatemia caused a gradual decline in the brain rate of glucose utilization in air-breathing animals to 50-70% of control rates. Results with both tracers were similar. Concentrations of Krebs cycle intermediates and glutamate did not decrease. These findings indicate that lactate can partially replace glucose as an oxidative fuel for developing rat brain. Hypercapnia depressed the rate of glucose utilization by developing brain and rates were 30-40% lower still in lactate-injected hypercapnic rats. Decreases in levels of Krebs cycle intermediates and glutamate were similar in both groups. Thus, lactate and CO2 are additive in their depressant effects on brain glucose utilization. The observation that lactate did not prevent the decreases in Krebs cycle intermediates and glutamate caused by hypercapnic acidosis suggests an inhibition of flux through pyruvate dehydrogenase during hypercapnia. The data from this study, coupled with data on lactate transport across the blood-brain barrier, indicate that the direction of movement of lactate and its rate of utilization by developing brain are functions of its concentration on blood relative to brain. Physiological and pathological conditions which elevate blood lactate levels above those in brain will, then, have a sparing effect upon brain glucose utilization.


Development and psychometric evaluation of the Perceived Social Stigma Questionnaire (PSSQ-for adults with epilepsy): A mixed method study.
Stigma is a subjective and multidimensional concept that is dependent on the understanding of individuals and social culture potentially influencing treatment and health in various aspects. The study of patient beliefs about epilepsy can help design preventive interventions. This study was conducted to develop and psychometrically evaluate the Perceived Social Stigma Questionnaire (PSSQ-for adults with epilepsy). This mixed method study of sequential exploratory quantitative-qualitative design was conducted within the framework of instrument development variant. The qualitative section consisted of item development and scale development. In the quantitative section (item analysis), the instrument was validated by investigating face, content, and construct validity and its reliability by internal consistency and stability. The PSSQ-for adults with epilepsy is a culturally appropriate measure with strong psychometric properties. The tool developed in this study can measure the perceived social stigma of epilepsy. However, more information on the scale is needed before it is adequately improved to test interventions, including concurrent validity and sensitivity to change.


Diazo coupling method for covalent attachment of proteins to solid substrates.
We describe a process for covalently linking proteins to glass microscope slides and microbeads in a manner that optimizes the reactivity of the immobilized proteins and that is suitable for high-throughput microarray and flow cytometry analysis. The method involves the diazo coupling of proteins onto activated self-assembled monolayers formed from p-aminophenyl trimethoxysilane. Proteins immobilized by this method maintained bioactivity and produced enhanced levels of protein-protein interaction, low background fluorescence, and high selectivity. The binding of immobilized proteins to their specific binding partner was analyzed quantitatively and successfully correlated with solution concentrations. Diazotized surfaces bound more efficiently to proteins containing a hexahistidine tag than those without a his-tag. Moreover, significantly higher reactivity of the immobilized his-tagged proteins was observed on diazotized surfaces than on amine-terminated surfaces. Results suggest that his-tagged proteins are immobilized by reaction of the his-tag with the diazotized surface, thus offering the possibility for preferential orientation of covalently bound proteins.


Conformation of circular DNA in two dimensions.
The conformation of circular DNA molecules of various lengths adsorbed in a 2D conformation on a mica surface is studied. The results confirm the conjecture that the critical exponent nu is topologically invariant and equal to the self-avoiding walk value (in the present case nu=3/4), and that the topology and dimensionality of the system strongly influence the crossover between the rigid regime and the self-avoiding regime at a scale L approximately 7l{p}. Additionally, the bond correlation function scales with the molecular length L as predicted. For molecular lengths L<or=5l{p}, circular DNA behaves like a stiff molecule with an approximately elliptic shape.


A prospective study of running injuries: the Vancouver Sun Run "In Training" clinics.
Seventeen running training clinics were investigated to determine the number of injuries that occur in a running programme designed to minimise the injury rate for athletes training for a 10 km race. The relative contributions of factors associated with injury were also reported. A total of 844 primarily recreational runners were surveyed in three trials on the 4th, 8th, and 12th week of the 13 week programme of the "In Training" running clinics. Participants were classified as injured if they experienced at least a grade 1 injury-that is, pain only after running. Logistic regression modelling and odds ratio calculation were performed for each sex using the following predictor variables: age, body mass index (BMI), previous aerobic activity, running frequency, predominant running surface, arch height, running shoe age, and concurrent cross training. Age played an important part in injury in women: being over 50 years old was a risk factor for overall injury, and being less than 31 years was protective against new injury. Running only one day a week showed a non-significant trend for injury risk in men and was a significant risk factor in women and overall injury. A BMI of > 26 kg/m(2) was reported as protective for men. Running shoe age also significantly contributed to the injury model. Half of the participants who reported an injury had had a previous injury; 42% of these reported that they were not completely rehabilitated on starting the 13 week training programme. An injury rate of 29.5% was recorded across all training clinics surveyed. The knee was the most commonly injured site. Although age, BMI, running frequency (days a week), and running shoe age were associated with injury, these results do not take into account an adequate measure of exposure time to injury, running experience, or previous injury and should thus be viewed accordingly. In addition, the reason for the discrepancy in injury rate between these 17 clinics requires further study.


Long-term effect of cognitive-behavioural therapy in patients with Hypochondriacal Disorder.
Cognitive-behavioural therapy (CBT) is an effective treatment for Hypochondriacal Disorder, but the long-term effect has not been examined extensively. To investigate the long-term effect of CBT on Hypochondriacal Disorder using several mental health measures. Follow-up time was at least 10 years. A total of 50 patients with a long history of Hypochondriacal Disorder, diagnosed according to ICD-10, received 16 sessions of individual CBT and were followed up with an uncontrolled design. All participants were assessed before and after the intervention period, and 10 years later. Intention-to-treat mixed-model repeated-measures analysis were conducted. The study has been registered at clinicaltrials.gov: NCT00959452. Patients displayed significant improvements across all outcomes, including level of health anxiety, somatisation, symptoms of anxiety and depression, quality of life, somatisation at treatment completion. Treatment gains were well maintained 10 years later. This uncontrolled treatment study suggests that patients treated with CBT for Hypochondriacal Disorder have significantly reduced health anxiety 1 year after treatment completion and the results are maintained 10 years later. The results indicate that CBT has a lasting effect, but the lack of a control group and use of only one therapist, means that care should be taken when generalising the findings.


Polymerase chain reaction for Streptococcus pyogenes used to evaluate an optical immunoassay for the detection of group A streptococci in children with pharyngitis.
In evaluations of sensitive rapid tests for group A streptococci such as the optical immunoassay (OIA), some samples are positive by the antigen test but negative by culture. A method is needed for resolving these discrepant results. To develop a PCR-based assay to detect group A streptococci and to use it to establish a reference standard for evaluating an OIA for group A streptococcal antigen. A PCR assay that detects a segment of the MF gene of Streptococcus pyogenes was developed for the detection of group A streptococci in throat swabs. Paired swabs were obtained from 200 children with symptomatic pharyngitis and used to perform OIA, agar culture, broth-enhanced culture and PCR. As a reference standard any patient with group A streptococci detected by either culture or PCR was considered to be truly positive. In comparison to agar and broth-enhanced culture procedures, OIA had sensitivities of 82 and 80% and specificities of 87 and 89%, respectively. Eight (44%) of 18 samples that were positive by OIA but negative by culture were positive for group A streptococci by PCR. Compared with the reference standard, sensitivities were OIA 76%, agar culture 79%, broth-enhanced culture 86% and PCR 96%. The specificity of OIA was 92%. PCR can be used to establish a reference standard for evaluating rapid tests for group A streptococci. With this reference standard OIA was nearly as sensitive as but less specific than agar culture for detection of group A streptococci. Maximum detection requires use of both tests.


[Incidence of metabolic syndrome and associated lifestyle factors in a worksite male population].
The prevention of metabolic syndrome (MetS) is one of the main targets of health promotion activities in the workplace. The present study aimed to clarify the incidence of MetS and associated lifestyle factors in a worksite male population. The study subjects consisted of 948 working men (mean age: 44 yr old) who did not meet the Japanese criteria for MetS during the annual health examination at a precision instrument development plant in Kanagawa, Japan, 2005. New-onset MetS was followed using the health examination data from 2006 to 2009. The incidence of MetS according to the combination of the presence of abdominal obesity and the number of other components of MetS (elevated blood pressure, dyslipidemia, and elevated fasting glucose) at baseline were calculated. The incidence of MetS was also compared among the groups with regard to differences in lifestyles (dietary habits, smoking status, sleep duration, exercise habits, and alcohol intake). A Cox proportional hazard model was used to identify independent factors contributing to an increased risk of MetS. New-onset MetS was observed in 76 subjects during a mean follow-up period of 3.7 yr. The incidence of MetS was 2.2/100 person-years and the 4-year incidence of MetS according to Kaplan-Meier analysis was 8.5%. The highest 4-year incidence of MetS was found in the group without abdominal obesity but with two or more components of MetS (37.9%) and the second highest incidence was found in the group with abdominal obesity and one other component of MetS (24.6%). The presence of abdominal obesity and each increase in the number of other components of MetS had an increased age-adjusted hazard ratio for an increased risk of MetS (5.23 and 4.79, respectively, both p<0.001). Similarly, sleep duration 5 h or less, current smoking, and ethanol intake 300 g/wk or more had an increased age-adjusted hazard ratio for an increased risk of MetS. The present study showed a high incidence of MetS in not only the group with abdominal obesity, but also the group without abdominal obesity but with two or more components of MetS. Sleep deprivation, current smoking, and excess alcohol intake were found to be factors contributing to an increased risk of MetS among several lifestyle factors examined. The usefulness of health promotion activities that preferentially target subjects who have such medical conditions and/or lifestyles are therefore expected to reduce the incidence of MetS in the workplace, from a high-risk strategy viewpoint.


The additional effect of hyaluronidase in lumbar interlaminar epidural injection.
To evaluate the effect of hyaluronidase in lumbar interlaminar epidural injection (LIEI) for low back pain and sciatica. Sixty-one patients suffering from severe low back pain and sciatica were randomly allocated into three groups. Group T (n=18, mean duration of illness: 2.12±1.16 months) received lumbar interlaminar epidural injection (LIEI) with 2 ml triamcinolone (40 mg/ml) and 5 ml bupivacaine (0.25%). Group H (n=16, mean duration of illness: 2.05±1.12 months) received LIEI with 1,500 IU hyaluronidase and 5 ml bupivacaine (0.25%). Group TH (n=27, mean duration of illness: 2.16±1.65 months) received LIEI with 1,500 IU hyaluronidase, 2 ml triamcinolone (40 mg/ml), and 5 ml bupivacaine (0.25%). The effects were evaluated using the Visual Analogue Scale (VAS) and Oswestry Disability Index (ODI) at preinjection and 2 weeks, 4 weeks, and 8 weeks after LIEI. Pain improved in all groups after 2 weeks (p<0.05). After 8 weeks, there was no significant difference in VAS improvement among the 3 groups. However, pain improved in 70.4% of Group TH compared with preinjection, in contrast to 44.4% of Group T and 31.3% of Group H. The ODI improved significantly only in Group TH after 8 weeks (p<0.05). LIEI with triamcinolone and hyaluronidase is more effective for reducing pain after 8 weeks than injection with triamcinolone or hyaluronidase alone.


Hormetic effect of panaxatriol saponins confers neuroprotection in PC12 cells and zebrafish through PI3K/AKT/mTOR and AMPK/SIRT1/FOXO3 pathways.
Hormesis is an adaptive response of living organisms to a moderate stress. However, its biomedical implication and molecular mechanisms remain to be intensively investigated. Panaxatriol saponins (PTS) is the major bioactive components extracted from Panax notoginseng, a widely used herbal medicine for cerebrovascular diseases. This study aims to examine the hormetic and neuroprotective effects of PTS in PC12 cells and zebrafish Parkinson's disease (PD) models. Our results demonstrated that PTS stimulated PC12 cell growth by about 30% at low doses, while PTS at high doses inhibited cell growth, which is a typical hormetic effect. Moreover, we found that low dose PTS pretreatment significantly attenuated 6-OHDA-induced cytotoxicity and up-regulated PI3K/AKT/mTOR cell proliferation pathway and AMPK/SIRT1/FOXO3 cell survival pathway in PC12 cells. These results strongly suggested that neuroprotective effects of PTS may be attributable to the hormetic effect induced by PTS through activating adaptive response-related signaling pathways. Notably, low dose PTS could significantly prevent the 6-OHDA-induced dopaminergic neuron loss and improve the behavior movement deficiency in zebrafish, whereas relative high dose PTS exhibited neural toxicity, further supporting the hormetic and neuroprotective effects of PTS. This study indicates that PTS may have the potential in the development of future therapeutic medicines for PD.


Combination therapy with local radiofrequency ablation and systemic vaccine enhances antitumor immunity and mediates local and distal tumor regression.
Radiofrequency ablation (RFA) is a minimally invasive energy delivery technique increasingly used for focal therapy to eradicate localized disease. RFA-induced tumor-cell necrosis generates an immunogenic source of tumor antigens known to induce antitumor immune responses. However, RFA-induced antitumor immunity is insufficient to control metastatic progression. We sought to characterize (a) the role of RFA dose on immunogenic modulation of tumor and generation of immune responses and (b) the potential synergy between vaccine immunotherapy and RFA aimed at local tumor control and decreased systemic progression. Murine colon carcinoma cells expressing the tumor-associated (TAA) carcinoembryonic antigen (CEA) (MC38-CEA(+)) were studied to examine the effect of sublethal hyperthermia in vitro on the cells' phenotype and sensitivity to CTL-mediated killing. The effect of RFA dose was investigated in vivo impacting (a) the phenotype and growth of MC38-CEA(+) tumors and (b) the induction of tumor-specific immune responses. Finally, the molecular signature was evaluated as well as the potential synergy between RFA and poxviral vaccines expressing CEA and a TRIad of COstimulatory Molecules (CEA/TRICOM). In vitro, sublethal hyperthermia of MC38-CEA(+) cells (a) increased cell-surface expression of CEA, Fas, and MHC class I molecules and (b) rendered tumor cells more susceptible to CTL-mediated lysis. In vivo, RFA induced (a) immunogenic modulation on the surface of tumor cells and (b) increased T-cell responses to CEA and additional TAAs. Combination therapy with RFA and vaccine in CEA-transgenic mice induced a synergistic increase in CD4(+) T-cell immune responses to CEA and eradicated both primary CEA(+) and distal CEA(-) s.c. tumors. Sequential administration of low-dose and high-dose RFA with vaccine decreased tumor recurrence compared to RFA alone. These studies suggest a potential clinical benefit in combining RFA with vaccine in cancer patients, and augment support for this novel translational paradigm.


Silibinin attenuates methotrexate-induced pulmonary injury by targeting oxidative stress.
The aim of the present study was to assess the protective effect of silibinin against methotrexate (MTX)-induced pulmonary toxicity. Rats were divided into four groups (MTX, MTX + silibinin, silibinin and control. MTX was injected intraperitoneally (i.p) into female Wistar rats (10 mg/kg/day for 3 days), which resulted in significant increases in the serum levels of alanine aminotransferase, aspartate aminotransferase and oxidant enzymes, including nitric oxide and myeloperoxidase. Furthermore, significant reductions were detected in the serum activity levels of the antioxidative enzymes, glutathione peroxidase and superoxide dismutase, when compared with the control group. However, administration of silibinin (100 mg/kg/day for 10 days, i.p.) was shown to ameliorate the MTX-induced pulmonary toxicity, as indicated by the normalization of the oxidative stress parameters. Furthermore, silibinin treatment was demonstrated to reduce the histopathological changes associated with MTX. In conclusion, silibinin exhibited protective effects against MTX-induced pulmonary toxicity, which may be attributed to its antioxidant activity.


Representation of the voice onset time (VOT) speech parameter in population responses within primary auditory cortex of the awake monkey.
Voice onset time (VOT) signifies the interval between consonant onset and the start of rhythmic vocal-cord vibrations. Differential perception of consonants such as /d/ and /t/ is categorical in American English, with the boundary generally lying at a VOT of 20-40 ms. This study tests whether previously identified response patterns that differentially reflect VOT are maintained in large-scale population activity within primary auditory cortex (A1) of the awake monkey. Multiunit activity and current source density patterns evoked by the syllables /da/ and /ta/ with variable VOTs are examined. Neural representation is determined by the tonotopic organization. Differential response patterns are restricted to lower best-frequency regions. Response peaks time-locked to both consonant and voicing onsets are observed for syllables with a 40- and 60-ms VOT, whereas syllables with a 0- and 20-ms VOT evoke a single response time-locked only to consonant onset. Duration of aspiration noise is represented in higher best-frequency regions. Representation of VOT and aspiration noise in discrete tonotopic areas of A1 suggest that integration of these phonetic cues occurs in secondary areas of auditory cortex. Findings are consistent with the evolving concept that complex stimuli are encoded by synchronized activity in large-scale neuronal ensembles.


Cross-correlation technique for dispersion characterization of chirped volume Bragg gratings.
We propose using cross-correlation frequency-resolved optical gating for dispersion characterization of optical elements with high dispersion, such as ultrashort pulse stretchers and compressors. The technique is based on spectrally resolved second-order cross correlation (sum frequency generation) of a stretched pulse with a reference short pulse. Dispersion of optical elements with a high pulse stretching ratio can be completely characterized using this method, even with moderate resolution of spectral measurements of the cross-correlation signal. The proposed technique is used to measure dispersion of a chirped Bragg grating recorded in photo-thermo-refractive glass. It was found that dispersion of these gratings is almost linear with wavelength and has approximately the same slope with opposite signs for two orientations of the grating with opposite faces of the grating used as the input face. Analysis of higher-order dispersion of the grating shows some variations of dispersion across the aperture of the grating, mostly in the amount of third-order dispersion contribution.


YouTube as a potential learning tool to help distinguish tonic-clonic seizures from nonepileptic attacks.
Medical students are increasingly turning to the website YouTube as a learning resource. This study set out to determine whether the videos on YouTube accurately depict the type of seizures that a medical student may search for. Two consultant epileptologists independently assessed the top YouTube videos returned following searches for eight terms relating to different categories of seizures. The videos were rated for their technical quality, concordance of diagnosis with an epileptologist-assigned diagnosis, and efficacy as a learning tool for medical education. Of the 200 videos assessed, 106 (63%) met the inclusion criteria for further analysis. Technical quality was generally good and only interfered with the diagnostic process in 8.5% of the videos. Of the included videos, 40.6-46.2% were judged to depict the purported diagnosis with moderate agreement between raters (75% agreement, κ=0.50). Of the videos returned after searching "tonic-clonic seizure", 28.6-35.7% were judged to show nonepileptic seizures with almost perfect interrater agreement (92.9% agreement, κ=0.84). Of the videos returned following the search "pseudoseizure", 77.8-88.9% of videos were judged to show nonepileptic seizures with substantial agreement (88.9% agreement, κ=0.61). Across all search terms, 19.8-33% of videos were judged as potentially useful as a learning resource, with fair agreement between raters (75.5% agreement, κ=0.38). These findings suggest that the majority of videos on YouTube claiming to show specific seizure subtypes are inaccurate, and YouTube should not be recommended as a learning tool for students. However, a small group of videos provides excellent demonstrations of tonic-clonic and nonepileptic seizures, which could be used by an expert teacher to demonstrate the difference between epileptic and nonepileptic seizures.


APS could potentially activate hepatic insulin signaling in HFD-induced IR mice.
Astragalus polysaccharide (APS) is the main component of Astragalus membranaceus, an anti-diabetic herb being used for thousands of years in Traditional Chinese medicine (TCM). In this study, we aimed to evaluate the impact of APS on hepatic insulin signaling, autophagy and ER stress response in high-fat-diet (HFD)-induced insulin resistance (IR) mice. APS was intra-gastrically administrated and metformin was used as a control medicine. Apart from monitoring the changes in the important parameters of IR progression, the gene and protein expression of the key factors marking the state of hepatic ER stress and autophagic flux were examined. We found that, largely comparable to the metformin regime, APS treatment resulted in an overall improvement of IR, as indicated by better control of body weight and blood glucose/lipid levels, recovery of liver functions and regained insulin sensitivity. In particular, the excessive and pro-apoptotic ER stress response and inhibition of autophagy, as a result of prolonged HFD exposure, were significantly corrected by APS administration, indicating a switch of the cellular fate in favor of cell survival. Using the HepG2/IR cell model, we demonstrated that APS modulated the insulin-initiated phosphorylation cascades in a similar manner to metformin. This study provides a rationale for exploiting the insulin-sensitizing potential of APS, which has a therapeutic performance almost equivalent to metformin, to enrich our options in the treatment of IR.


Mouse models of heart failure: cell signaling and cell survival.
Heart failure is one of the paramount global causes of morbidity and mortality. Despite this pandemic need, the available clinical counter-measures have not altered substantially in recent decades, most notably in the context of pharmacological interventions. Cell death plays a causal role in heart failure, and its inhibition poses a promising approach that has not been thoroughly explored. In previous approaches to target discovery, clinical failures have reflected a deficiency in mechanistic understanding, and in some instances, failure to systematically translate laboratory findings toward the clinic. Here, we review diverse mouse models of heart failure, with an emphasis on those that identify potential targets for pharmacological inhibition of cell death, and on how their translation into effective therapies might be improved in the future.


Comparison of biopsy-based methods for the detection of Helicobacter pylori infection.
Various biopsy-based methods for the detection of Helicobacter pylori are evaluated to determine their sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV), followed by polymerase chain reaction (PCR) for the 16S ribosomal RNA (rRNA) gene of H. pylori (16S PCR) to confirm the results. Seventyfive patients (65% [49] males, age range: 17-77 years, mean 42+/-14.6 years) with dyspeptic symptoms are included in the study. Gastric antrum biopsy specimens collected during endoscopy are tested using a urea agar base enriched with 40% urea solution (eUAB, Oxoid)), a commercial rapid urease test (Pronto Dry, Medical Instrument Corp, Switzerland), histopathology and 16S PCR. The eUAB test showed 97% sensitivity, 86% specificity, 84% PPV, 97% NPV and 91% accuracy when the diagnosis of H. pylori infection was made with positive Pronto Dry and histopathology. Pronto Dry showed 100% sensitivity, 82% specificity, 80% PPV, 100% NPV and 89% accuracy when the diagnosis of H. pylori infection was made on positive histopathology and eUAB. Thus, the eUAB can be used as a rapid urease test. It is economical and has a sensitivity and specificity comparable to a commercially available rapid urease test to detect urease activity of H. pylori in gastric biopsy.


Molecular differentiation of Treponema pallidum subspecies in skin ulceration clinically suspected as yaws in Vanuatu using real-time multiplex PCR and serological methods.
We developed a TaqMan-based real-time quadriplex polymerase chain reaction (PCR) to simultaneously detect Treponema pallidum subspecies pallidum, T. pallidum subsp. pertenue, and T. pallidum subsp. endemicum, the causative agents of venereal syphilis, yaws, and bejel, respectively. The PCR assay was applied to samples from skin ulcerations of clinically presumptive yaws cases among children on Tanna Island, Vanuatu. Another real-time triplex PCR was used to screen for the point mutations in the 23S rRNA genes that have previously been associated with azithromycin resistance in T. pallidum subsp. pallidum strains. Seropositivity by the classical syphilis serological tests was 35.5% among children with skin ulcerations clinically suspected with yaws, whereas the presence of T. pallidum subsp. pertenue DNA was only found in lesions from 15.5% of children. No evidence of T. pallidum subsp. pertenue infection, by either PCR or serology was found in ∼59% of cases indicating alternative causes of yaws-like lesions in this endemic area.


An anti-inflammatory role for interleukin-11 in established murine collagen-induced arthritis.
Interleukin-11 (IL-11) is a cytokine belonging to the IL-6 family which has both pro- and anti-inflammatory potential. Like IL-6 it can diminish tumour necrosis factor-alpha and IL-1 production, and augment immunoglobulin synthesis. We have explored the immunomodulatory effects of IL-11 treatment in mice in a model of inflammatory autoimmune joint disease, collagen-induced arthritis (CIA). Recombinant human IL-11 was administered at various doses to DBA/1 mice after the onset of CIA. IL-11 treatment caused a significant reduction in the clinical severity of established CIA, which was associated with protection from joint damage, as assessed by histology. Although there was a suggestion at high doses of IL-11 that the anticollagen type II (CII) response may have been augmented, there was no statistically significant effect of IL-11 treatment on anti-CII antibody levels. Similarly, the acute-phase reactant serum amyloid P was only elevated in mice receiving very high doses (50-100 microgram/day) of IL-11. Endogenous IL-11 was abundantly produced in synovial membrane cultures derived from CII-immunized mice with active disease, suggesting that, as in rheumatoid arthritis, this cytokine is spontaneously produced in the inflammatory response in CIA. The results presented here demonstrate an anti-arthritic immunoregulatory role for IL-11 in murine CIA, and suggest that IL-11 is a candidate therapeutic molecule for human inflammatory arthritic diseases.


External-energy-independent polymer capsule motors and their cooperative behaviors.
The design and development of mobile nano-, micro-, and millimeter-scale autonomous systems have been perused over several decades. Here, we introduce a millimeter-sized polymer capsule motor with specific features and functionalities. It runs without any external energy sources or the consumption of external fuels such as H(2)O(2) or glucose. The occurrence of motion is due to the asymmetric release of organic solvent from the capsule and the asymmetric change in the surface tension of the surrounding liquid. The capsule moves from a place of lower surface tension to a place of higher surface tension (Marangoni effect) in an attempt by the system to attain the desirable lowest-free-energy state. The operation of the motor is versatile in terms of the environment, as it moves on a wide variety of liquid/air interfaces, including water, sea water, organic solvent/water mixtures, and acids. A high-motion velocity was observed, with a travelling distance of over 20 meters. The manipulation of its motion was achieved upon functionalization with nickel powder and application of an external magnetic field. Long-range interaction behaviors and surface-cleaning effects due to the chemotaxis effect were also demonstrated when the capsule was functionalized with sodium dodecyl sulfate (SDS). We believe that a plethora of applications can be envisioned with this motor, such as cargo delivery, manipulation of matter, sensing and detection, biorecognition, and environmental remediation.


Normative functional fitness standards and trends of Portuguese older adults: cross-cultural comparisons.
This cross-sectional study was designed to develop normative functional fitness standards for the Portuguese older adults, to analyze age and gender patterns of decline, to compare the fitness level of Portuguese older adults with that of older adults in other countries, and to evaluate the fitness level of Portuguese older adults relative to recently published criterion fitness standards associated with maintaining physical independence. A sample of 4,712 independent-living older adults, age 65-103 yr, was evaluated using the Senior Fitness Test battery. Age-group normative fitness scores are reported for the 10th, 25th, 50th, 75th, and 90th percentiles. Results indicate that both women and men experience age-related losses in all components of functional fitness, with their rate of decline being greater than that observed in other populations, a trend which may cause Portuguese older adults to be at greater risk for loss of independence in later years. These newly established normative standards make it possible to assess individual fitness level and provide a basis for implementing population-wide health strategies to counteract early loss of independence.


Effectiveness of strengthening and stretching exercises for the postural correction of abducted scapulae: a review.
Abnormal postural alignment can be detrimental to muscle function, is aesthetically unpleasing, and might contribute to joint pain. It has been unclear as to whether stretching or strengthening exercises can correct faulty posture such as abducted scapulae. It has been postulated that short and tight scapular abductor muscles or weak and lengthened scapular retractor muscles or a combination cause an abducted scapulae posture and that exercise can correct this condition. The purpose of this review was to compile the information on factors influencing scapular position at rest, examine the effectiveness of exercise interventions in altering scapular position, and make recommendations for future research. When examining the different methods that have been used to determine the position of the scapula, attention should be paid to their respective reliability and validity. Correlational studies have failed to detect a significant association between muscle strength and scapular position but found a significant relationship between muscle length and scapular position. Prospective intervention studies have shown that stretching the anterior chest muscles on its own or in combination with strengthening the scapular retractors can alter the position of the scapula at rest in individuals with abducted scapulae. Although these results are encouraging, there is a dearth of high-quality studies and more research is required to address the limitations of the studies. None of the intervention studies measured strength or flexibility pre or post intervention, so it is unclear how effective the intervention was in changing these factors and the actual mechanism behind the change. To determine which component of the intervention is most effective and whether the results are additive, future research should include stretching only, strengthening only, and combined stretching and strengthening groups. Follow-up measurements at some period after completion of the intervention would also provide important information as to the permanency of any changes. The practical implication is that caution must be displayed when considering the promotion of strengthening exercises to try and correct for abnormal scapular posture until further evidence becomes available.


A Potential Golden Age to Come-Current Tools, Recent Use Cases, and Future Avenues for De Novo Sequencing in Proteomics.
In shotgun proteomics, peptide and protein identification is most commonly conducted using database search engines, the method of choice when reference protein sequences are available. Despite its widespread use the database-driven approach is limited, mainly because of its static search space. In contrast, de novo sequencing derives peptide sequence information in an unbiased manner, using only the fragment ion information from the tandem mass spectra. In recent years, with the improvements in MS instrumentation, various new methods have been proposed for de novo sequencing. This review article provides an overview of existing de novo sequencing algorithms and software tools ranging from peptide sequencing to sequence-to-protein mapping. Various use cases are described for which de novo sequencing was successfully applied. Finally, limitations of current methods are highlighted and new directions are discussed for a wider acceptance of de novo sequencing in the community.


Marginal Structural Models to Assess Delays in Second-Line HIV Treatment Initiation in South Africa.
South African HIV treatment guidelines call for patients who fail first-line antiretroviral therapy (ART) to be switched to second-line ART, yet logistical issues, clinician decisions and patient preferences make delay in switching to second-line likely. We explore the impact of delaying second-line ART after first-line treatment failure on rates of death and virologic failure. We include patients with documented virologic failure on first-line ART from an observational cohort of 9 South African clinics. We explored predictors of delayed second-line switch and used marginal structural models to analyze rates of death following first-line failure by categorical time to switch to second-line. Cox proportional hazards models were used to examine virologic failure on second-line ART among patients who switched to second-line. 5895 patients failed first-line ART, and 63% switched to second-line. Among patients who switched, median time to switch was 3.4 months (IQR: 1.1-8.7 months). Longer time to switch was associated with higher CD4 counts, lower viral loads and more missed visits prior to first-line failure. Worse outcomes were associated with delay in second-line switch among patients with a peak CD4 count on first-line treatment ≤100 cells/mm3. Among these patients, marginal structural models showed increased risk of death (adjusted HR for switch in 6-12 months vs. 0-1.5 months = 1.47 (95% CI: 0.94-2.29), and Cox models showed increased rates of second-line virologic failure despite the presence of survivor bias (adjusted HR for switch in 3-6 months vs. 0-1.5 months = 2.13 (95% CI: 1.01-4.47)). Even small delays in switch to second-line ART were associated with increased death and second-line failure among patients with low CD4 counts on first-line. There is opportunity for healthcare providers to switch patients to second-line more quickly.


Phase retrieval with transverse translation diversity: a nonlinear optimization approach.
We develop and test a nonlinear optimization algorithm for solving the problem of phase retrieval with transverse translation diversity, where the diverse far-field intensity measurements are taken after translating the object relative to a known illumination pattern. Analytical expressions for the gradient of a squared-error metric with respect to the object, illumination and translations allow joint optimization of the object and system parameters. This approach achieves superior reconstructions, with respect to a previously reported technique [H. M. L. Faulkner and J. M. Rodenburg, Phys. Rev. Lett. 93, 023903 (2004)], when the system parameters are inaccurately known or in the presence of noise. Applicability of this method for samples that are smaller than the illumination pattern is explored.


Emerging concepts in immunity to hepatitis C virus infection.
Since the discovery of hepatitis C virus (HCV) by molecular cloning almost a quarter of a century ago, unprecedented at the time because the virus had never been grown in cell culture or detected serologically, there have been impressive strides in many facets of our understanding of the natural history of the disease, the viral life cycle, the pathogenesis, and antiviral therapy. It is apparent that the virus has developed multiple strategies to evade immune surveillance and eradication. This Review covers what we currently understand of the temporal and spatial immunological changes within the human innate and adaptive host immune responses that ultimately determine the outcomes of HCV infection.


Influence of NaCl on shape deformation of polymersomes.
Polymersomes frequently appear in the literature as promising candidates for a wide range of applications from targeted drug delivery to nanoreactors. From a cell mimetic point of view, it is important to understand the size and shape changes of the vesicles in the physiological environment since that can influence the drug delivery mechanism. In this work we studied the structural features of polymersomes consisting of poly(ethylene glycol)-poly(dimethylsiloxane)-poly(ethylene glycol) at the nanoscopic length scale in the presence of NaCl, which is a very common molecule in the biotic aqueous environment. We used dynamic light scattering (DLS), cryo-TEM, small angle neutron scattering (SANS) and small angle X-ray scattering (SAXS). We observed transformation of polymersomes from spherical to elongated vesicles at low salt concentration and into multivesicular structures at high salt concentration. Model fitting analysis of SANS data indicated a reduction of vesicle radius up to 47% and from the SAXS data we observed an increase in membrane thickness up to 8% and an increase of the PDMS hydrophobic segment up to 11% indicating stretching of the membrane due to osmotic imbalance. Also, from the increase in the interlamellar repeat distance up to 98% under high salt concentrations, we concluded that the shape and structural changes observed in the polymersomes are a combined result of osmotic pressure change and ion-membrane interactions.


Synthetic lethal screen identifies NF-κB as a target for combination therapy with topotecan for patients with neuroblastoma.
Despite aggressive multimodal treatments the overall survival of patients with high-risk neuroblastoma remains poor. The aim of this study was to identify novel combination chemotherapy to improve survival rate in patients with high-risk neuroblastoma. We took a synthetic lethal approach using a siRNA library targeting 418 apoptosis-related genes and identified genes and pathways whose inhibition synergized with topotecan. Microarray analyses of cells treated with topotecan were performed to identify if the same genes or pathways were altered by the drug. An inhibitor of this pathway was used in combination with topotecan to confirm synergism by in vitro and in vivo studies. We found that there were nine genes whose suppression synergized with topotecan to enhance cell death, and the NF-κB signaling pathway was significantly enriched. Microarray analysis of cells treated with topotecan revealed a significant enrichment of NF-κB target genes among the differentially altered genes, suggesting that NF-κB pathway was activated in the treated cells. Combination of topotecan and known NF-κB inhibitors (NSC 676914 or bortezomib) significantly reduced cell growth and induced caspase 3 activity in vitro. Furthermore, in a neuroblastoma xenograft mouse model, combined treatment of topotecan and bortezomib significantly delayed tumor formation compared to single-drug treatments. Synthetic lethal screening provides a rational approach for selecting drugs for use in combination therapy and warrants clinical evaluation of the efficacy of the combination of topotecan and bortezomib or other NF-κB inhibitors in patients with high risk neuroblastoma.


Ion mobility spectrometry-mass spectrometry defines the oligomeric intermediates in amylin amyloid formation and the mode of action of inhibitors.
The molecular mechanisms by which different proteins assemble into highly ordered fibrillar deposits and cause disease remain topics of debate. Human amylin (also known as islet amyloid polypeptide/hIAPP) is found in vivo as amyloid deposits in the pancreatic islets of sufferers of type II diabetes mellitus, and its self-aggregation is thought to be a pathogenic factor in disease and to contribute to the failure of islet transplants. Here, electrospray ionization-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS) has been used to monitor oligomer formation from IAPP. The detection, identification and characterization of oligomers from both human and rat amylin (rIAPP) are described. Oligomers up to and including hexamers have been detected for both peptides. From ESI-IMS-MS derived collision cross sections (CCS), these species are shown to be elongated in conformation. Collision-induced dissociation (CID-MS/MS) revealed differences in the gas-phase stability of the oligomers formed from hIAPP and rIAPP, which may contribute to their differences in amyloid propensity. Using ESI-IMS-MS, the mode of inhibition of amyloid formation from hIAPP using small molecules or co-incubation with rIAPP was also investigated. We show that the polyphenolic compounds epigallocatechin gallate (EGCG) and silibinin bind to specific conformers within a dynamic ensemble of hIAPP monomers, altering the progress of oligomerization and fibril assembly. Hetero-oligomer formation also occurs with rIAPP but leads only to inefficient inhibition. The results indicate that although different small molecules can be effective inhibitors of hIAPP self-assembly, their modes of action are distinct and can be distinguished using ESI-IMS-MS.


DNMT3A(R882H) mutant and Tet2 inactivation cooperate in the deregulation of DNA methylation control to induce lymphoid malignancies in mice.
TEN-ELEVEN-TRANSLOCATION-2 (TET2) and DNA-METHYLTRANSFERASE-3A (DNMT3A), both encoding proteins involved in regulating DNA methylation, are mutated in hematological malignancies affecting both myeloid and lymphoid lineages. We previously reported an association of TET2 and DNMT3A mutations in progenitors of patients with angioimmunoblastic T-cell lymphomas (AITL). Here, we report on the cooperative effect of Tet2 inactivation and DNMT3A mutation affecting arginine 882 (DNMT3A(R882H)) using a murine bone marrow transplantation assay. Five out of eighteen primary recipients developed hematological malignancies with one mouse developing an AITL-like disease, two mice presenting acute myeloid leukemia (AML)-like and two others T-cell acute lymphoblastic leukemia (T-ALL)-like diseases within 6 months following transplantation. Serial transplantations of DNMT3A(R882H) Tet2(-/-) progenitors led to a differentiation bias toward the T-cell compartment, eventually leading to AITL-like disease in 9/12 serially transplanted recipients. Expression profiling suggested that DNMT3A(R882H) Tet2(-/-) T-ALLs resemble those of NOTCH1 mutant. Methylation analysis of DNMT3A(R882H) Tet2(-/-) T-ALLs showed a global increase in DNA methylation affecting tumor suppressor genes and local hypomethylation affecting genes involved in the Notch pathway. Our data confirm the transformation potential of DNMT3A(R882H) Tet2(-/-) progenitors and represent the first cooperative model in mice involving Tet2 inactivation driving lymphoid malignancies.


Parenting Style and Behavior as Longitudinal Predictors of Adolescent Alcohol Use.
Adolescent alcohol use is a serious problem in Australia and other nations. Longitudinal data on family predictors are valuable to guide parental education efforts. The present study tested Baumrind's proposal that parenting styles are direct predictors of adolescent alcohol use. Latent class modeling was used to investigate adolescent perceptions of parenting styles and multivariate regression to examine their predictive effect on the development of adolescent alcohol use. The data set comprised 2,081 secondary school students (55.9% female) from metropolitan Melbourne, Australia, who completed three waves of annual longitudinal data starting in 2004. Baumrind's parenting styles were significant predictors in unadjusted analyses, but these effects were not maintained in multivariate models that also included parenting behavior dimensions. Family influences on the development of adolescent alcohol use appear to operate more directly through specific family management behaviors rather than through more global parenting styles.


The development and validation of an urbanicity scale in a multi-country study.
Although urban residence is consistently identified as one of the primary correlates of non-communicable disease in low- and middle-income countries, it is not clear why or how urban settings predispose individuals and populations to non-communicable disease (NCD), or how this relationship could be modified to slow the spread of NCD. The urban-rural dichotomy used in most population health research lacks the nuance and specificity necessary to understand the complex relationship between urbanicity and NCD risk. Previous studies have developed and validated quantitative tools to measure urbanicity continuously along several dimensions but all have been isolated to a single country. The purposes of this study were 1) To assess the feasibility and validity of a multi-country urbanicity scale; 2) To report some of the considerations that arise in applying such a scale in different countries; and, 3) To assess how this scale compares with previously validated scales of urbanicity. Household and community-level data from the Young Lives longitudinal study of childhood poverty in 59 communities in Ethiopia, India and Peru collected in 2006/2007 were used. Household-level data include parents' occupations and education level, household possessions and access to resources. Community-level data include population size, availability of health facilities and types of roads. Variables were selected for inclusion in the urbanicity scale based on inspection of the data and a review of literature on urbanicity and health. Seven domains were constructed within the scale: Population Size, Economic Activity, Built Environment, Communication, Education, Diversity and Health Services. The scale ranged from 11 to 61 (mean 35) with significant between country differences in mean urbanicity; Ethiopia (30.7), India (33.2), Peru (39.4). Construct validity was supported by factor analysis and high corrected item-scale correlations suggest good internal consistency. High agreement was observed between this scale and a dichotomized version of the urbanicity scale (Kappa 0.76; Spearman's rank-correlation coefficient 0.84 (p < 0.0001). Linear regression of socioeconomic indicators on the urbanicity scale supported construct validity in all three countries (p < 0.05). This study demonstrates and validates a robust multidimensional, multi-country urbanicity scale. It is an important step on the path to creating a tool to assess complex processes like urbanization. This scale provides the means to understand which elements of urbanization have the greatest impact on health.


Preparation of thyroid follicular cells for mRNA quantification after fluorescence-activated cell sorting.
We established a novel method to analyze cells collected by fluorescence-activated cell sorting (FACS) named mRNA quantification after FACS (FACS-mQ) in which cells are labeled with a fluorescence dye in a manner that minimizes RNA degradation, and then cells sorted by FACS are examined by analyzing their gene expression profile. In order to analyze cells using FACS-mQ, it is essential to prepare single-cell suspensions without RNA degradation. We found that a new tissue preservation medium, ThelioKeep™, which contains epigallocatechin-3-gallate (EGCG), was suitable for preservation of thyroid tissues. The aim of this study was to establish a cell dispersion method of thyroid follicular cells using ThelioKeep™. We compared the efficiency of cell dispersion between the two methods, the conventional cold pre-incubation method and the ThelioKeep™ method; then we determined if cells obtained by the ThelioKeep™ method were suitable for FACS-mQ analysis. We found that a larger number of cells were recovered using ThelioKeep™ than using the conventional cold pre-incubation method. Furthermore, cell viability was higher with the ThelioKeep™ method than with the cold pre-incubation method. Thyroid cells collected by this method were analyzed by FACS-mQ. A clear shift in flow cytometry analysis was observed when cells were stained with an anti-thyroglobulin or anti-thyroid transcription factor-1 antibody. After sorting, the same copy number of ACTB mRNA was detected in thyroid cells as in an anaplastic carcinoma cell line, 8305C. These findings imply that preparation of thyroid cells using the present method is suitable for FACS-mQ analysis.


Separating twin images and locating the center of a microparticle in dense suspensions using correlations among reconstructed fields of two parallel holograms.
This paper deals with two issues affecting the application of digital holographic microscopy (DHM) for measuring the spatial distribution of particles in a dense suspension, namely discriminating between real and virtual images and accurate detection of the particle center. Previous methods to separate real and virtual fields have involved applications of multiple phase-shifted holograms, combining reconstructed fields of multiple axially displaced holograms, and analysis of intensity distributions of weakly scattering objects. Here, we introduce a simple approach based on simultaneously recording two in-line holograms, whose planes are separated by a short distance from each other. This distance is chosen to be longer than the elongated trace of the particle. During reconstruction, the real images overlap, whereas the virtual images are displaced by twice the distance between hologram planes. Data analysis is based on correlating the spatial intensity distributions of the two reconstructed fields to measure displacement between traces. This method has been implemented for both synthetic particles and a dense suspension of 2 μm particles. The correlation analysis readily discriminates between real and virtual images of a sample containing more than 1300 particles. Consequently, we can now implement DHM for three-dimensional tracking of particles when the hologram plane is located inside the sample volume. Spatial correlations within the same reconstructed field are also used to improve the detection of the axial location of the particle center, extending previously introduced procedures to suspensions of microscopic particles. For each cross section within a particle trace, we sum the correlations among intensity distributions in all planes located symmetrically on both sides of the section. This cumulative correlation has a sharp peak at the particle center. Using both synthetic and recorded particle fields, we show that the uncertainty in localizing the axial location of the center is reduced to about one particle's diameter.


Gold-doped silver nanocluster [Au3Ag38(SCH2Ph)24X5]2- (X = Cl or Br).
Here we report the single-crystal structure, experimental and theoretical characterization of a 41-metal atom Au-Ag alloy nanocluster [Au3Ag38(SCH2Ph)24X5]2- (1, X = Cl or Br). The nanocluster 1 is co-protected by thiolate and halogen atoms and features an all-metallic face-fused biicosahedral Au2@AuAg20 rod-like kernel enwrapped by the outermost Ag18(SCH2Ph)24X3 shell. Two sites on the surface of the biicosahedral kernel are partially occupied by Au and Ag atoms. The outer Ag18(SCH2Ph)24X3 shell is composed of two Ag6S6 cycles at the two poles and one Ag6S2X3 arc at the equator with both 2- and 3-coordinated Ag atoms, which has not been observed in gold or silver nanoclusters ever before. Theoretical calculations elucidate its electronic structure as well as optical properties, thus producing informative correlations between its structure and properties. This nanocluster exhibits near-infrared (NIR) emission around 825 nm. This work (i) snapshots a rare crystal structure of an Au-doped silver alloyed nanocluster; (ii) gives a deep insight to understand how the capping ligand or anions affect the structure of the alloy nanocluster; and (iii) provides precise information about gold atom doping site that is very significant in the recognition of potential active catalytic sites of the alloy nanoparticles.


The utility of next-generation sequencing in diagnosis and monitoring of acute myeloid leukemia and myelodysplastic syndromes.
Myeloid malignancies including acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) are a heterogeneous group of disorders that share a common biology and are a major source of morbidity and mortality. In the last several years, studies using next-generation sequencing (NGS) have identified a core set of recurrently mutated myeloid malignancy genes in the majority of patients with AML and MDS, including those with normal cytogenetics. DNA-level mutations in several of these genes including NPM1, FLT3, and CEBPA in AML and ASXL1, ETV6, EZH2, RUNX1, and TP53 in MDS are associated with changes in patient outcomes and are now tested for in clinical laboratories. In addition to providing prognostic information, these gene mutations can be used to monitor patient disease burden through the use of ultrasensitive detection techniques. In this review, we will focus on the clinical utility of various NGS-based methods including whole-genome sequencing, exome sequencing, and targeted panel-based sequencing in the initial diagnosis and management of AML and MDS and cover recent methodological advances for the molecular monitoring of AML and MDS.


A randomised controlled trial of intervention site radiotherapy in malignant pleural mesothelioma.
To assess the effectiveness of radiotherapy in preventing tumour seeding after chest drain or pleural biopsy in patients with malignant mesothelioma and to determine, if tract metastases appear, whether they are tender or troublesome to patients. Patients with a histological diagnosis of pleural mesothelioma and an invasive procedure within the preceding 21 days were stratified by age, performance status and treatment centre. Randomisation was performed between immediate drain site radiotherapy 21Gy in three fractions (XRT arm) or best supportive care (BSC) with follow-up to 12 months. Patients were asked to complete questionnaires on treatment toxicity and on symptoms from any tract metastases detected. Sixty-one patients were recruited from two centres between 1998 and 2004; 56 men, 5 women, median age 70. 31 were allocated to drain site radiotherapy. Seven patients developed tract metastases associated with the drain site (four XRT arm, three BSC) and four developed metastases associated with subsequent procedures at other sites (three XRT, one BSC). Two patients each developed two tract metastases. Of the 12 metastases, nine overlay the previous drain site but three were adjacent to the site. No statistically significant difference was found in the risk of tract metastasis associated with the drain site between the arms (p=0.748). Prophylactic drain site radiotherapy in malignant pleural mesothelioma does not reduce the incidence of tumour seeding by the margin indicated by previous studies.


Baclofen therapy for chronic hiccup.
Chronic hiccup is a rare but potentially severe condition, that can be symptomatic of a variety of diseases, or idiopathic. Many therapeutic interventions have been reported, most often as case reports. Among other drugs, baclofen has been suggested as a therapy for chronic hiccup. In a large series of patients, we have evaluated its therapeutic position. In patients with chronic hiccup, defined as hiccup spell or recurring hiccup attacks lasting more than 7 days, investigation of the upper gastro-oesophageal tract (fibroscopy, manometry, and pH monitoring) was systematically performed. Most patients had tried numerous drugs in the past, without success. Baclofen was used as a first treatment in patients without evidence of any gastro-oesophageal disease (n = 17), and was undertaken only after full treatment of such disease (n = 55) had failed to solve the hiccup problem (n = 20). Baclofen has, therefore, been administered to 37 patients with chronic hiccup (average duration 4.6 yrs). Baclofen produced a long-term complete resolution (18 cases) or a considerable decrease (10 cases) of hiccups in 28 of the 37 patients. There was no significant difference between patients with or without gastro-oesophageal disease. We conclude that so-called idiopathic chronic hiccup often results from gastro-oesophageal abnormalities. Also, if controlled studies confirm our encouraging results, baclofen can be a major element in the treatment of chronic hiccup that is idiopathic, or that cannot be helped by treatment of gastro-oesophageal diseases.


Improved gait recognition by gait dynamics normalization.
Potential sources for gait biometrics can be seen to derive from two aspects: gait shape and gait dynamics. We show that improved gait recognition can be achieved after normalization of dynamics and focusing on the shape information. We normalize for gait dynamics using a generic walking model, as captured by a population Hidden Markov Model (pHMM) defined for a set of individuals. The states of this pHMM represent gait stances over one gait cycle and the observations are the silhouettes of the corresponding gait stances. For each sequence, we first use Viterbi decoding of the gait dynamics to arrive at one dynamics-normalized, averaged, gait cycle of fixed length. The distance between two sequences is the distance between the two corresponding dynamics-normalized gait cycles, which we quantify by the sum of the distances between the corresponding gait stances. Distances between two silhouettes from the same generic gait stance are computed in the linear discriminant analysis space so as to maximize the discrimination between persons, while minimizing the variations of the same subject under different conditions. The distance computation is constructed so that it is invariant to dilations and erosions of the silhouettes. This helps us handle variations in silhouette shape that can occur with changing imaging conditions. We present results on three different, publicly available, data sets. First, we consider the HumanlD Gait Challenge data set, which is the largest gait benchmarking data set that is available (122 subjects), exercising five different factors, i.e., viewpoint, shoe, surface, carrying condition, and time. We significantly improve the performance across the hard experiments involving surface change and briefcase carrying conditions. Second, we also show improved performance on the UMD gait data set that exercises time variations for 55 subjects. Third, on the CMU Mobo data set, we show results for matching across different walking speeds. It is worth noting that there was no separate training for the UMD and CMU data sets.


Human immunodeficiency virus type 1 reverse transcriptase. 3'-Azidodeoxythymidine 5'-triphosphate inhibition indicates two-step binding for template-primer.
Human immunodeficiency virus type-1 (HIV-1) reverse transcriptase (RT) catalyzes DNA synthesis by an ordered sequential mechanism. After template-primer (T.P) binds to free enzyme, the deoxynucleoside triphosphate to be incorporated binds to the RT and T.P binary complex (RTT.P). After incorporation of the bound nucleotide, catalytic cycling is limited either by a conformational change (for processive synthesis) or release of the enzyme from the extended T.P (for single-nucleotide incorporation). To explore cycling through these alternate rate-limiting steps, we determined kinetic parameters for single-nucleotide incorporation by HXB2R HIV-1 RT with chain-terminating nucleotide substrates 3'-azido-3'-deoxythymidine triphosphate (AZTTP) and dideoxythymidine triphosphate on a homopolymeric T.P system, poly(rA)-oligo(dT)16. Inhibition of processive deoxythymidine monophosphate incorporation by these chain-terminating substrates was also examined. Because AZTTP is a substrate, its Km should be equivalent to Ki, and since Km for AZTTP should be influenced by the dissociation rate constant for RTT.P, we examined the effect of altering RTT.P dissociation on AZTTP kinetic parameters. The dissociation rate constant was modulated by making use of different T.P substrates, viral sources of RT, and a mutant RT altered at a residue that perturbs T.P binding. As expected from earlier work, the time course of AZTMP incorporation on poly(rA)-oligo(dT)16 was biphasic, with a burst followed by a slower steady-state phase representing kcat (0.42 min-1) which was similar to the rate constant for RTT.P dissociation. Additionally, Km for AZTTP (110 nM) was lower than its equilibrium dissociation constant (1200 nM). AZTTP inhibition (Ki,AZTTP) of processive dTMP incorporation and incorporation of a single nucleotide were similar. However, a simple correlation between the RTT.P dissociation rate constant and Ki,AZTTP was not observed. These results indicate that a simple ordered model for single-nucleotide incorporation is inadequate and that different forms of RTT.P exist which can limit catalysis. The results are discussed in the context of a two-step binding reaction for T.P where the binary RTT.P complex undergoes an isomerization before binding of the deoxynucleotide substrate.


Using BCG vaccine to enhance non-specific protection of health care workers during the COVID-19 pandemic: A structured summary of a study protocol for a randomised controlled trial in Denmark.
The Bacille Calmette-Guérin (BCG) vaccine against tuberculosis is associated with non- specific protective effects against other infections, and significant reductions in all-cause morbidity and mortality have been reported. We aim to test whether BCG vaccination may reduce susceptibility to and/or the severity of COVID-19 and other infectious diseases in health care workers (HCW) and thus prevent work absenteeism.The primary objective is to reduce absenteeism due to illness among HCW during the COVID-19 pandemic. The secondary objectives are to reduce the number of HCW that are infected with SARS-CoV-2, and to reduce the number of hospital admissions among HCW during the COVID-19 pandemic. BCG vaccination of HCW will reduce absenteeism by 20% over a period of 6 months. Placebo-controlled, single-blinded, randomised controlled trial, recruiting study participants at several geographic locations. The BCG vaccine is used in this study on a different indication than the one it has been approved for by the Danish Medicines Agency, therefore this is classified as a phase III study. The trial will recruit 1,500 HCW at Danish hospitals.To be eligible for participation, a subject must meet the following criteria: Adult (≥18 years); Hospital personnel working at a participating hospital for more than 22 hours per week.A potential subject who meets any of the following criteria will be excluded from participation in this study: Known allergy to components of the BCG vaccine or serious adverse events to prior BCG administration Known prior active or latent infection with Mycobacterium tuberculosis (M. tuberculosis) or other mycobacterial species Previous confirmed COVID-19 Fever (>38 C) within the past 24 hours Suspicion of active viral or bacterial infection Pregnancy Breastfeeding Vaccination with other live attenuated vaccine within the last 4 weeks Severely immunocompromised subjects. This exclusion category comprises: a) subjects with known infection by the human immunodeficiency virus (HIV-1) b) subjects with solid organ transplantation c) subjects with bone marrow transplantation d) subjects under chemotherapy e) subjects with primary immunodeficiency f) subjects under treatment with any anti-cytokine therapy within the last year g) subjects under treatment with oral or intravenous steroids defined as daily doses of 10 mg prednisone or equivalent for longer than 3 months h) Active solid or non-solid malignancy or lymphoma within the prior two years Direct involvement in the design or the execution of the BCG-DENMARK-COVID trial Intervention and comparator: Participants will be randomised to BCG vaccine (BCG-Denmark, AJ Vaccines, Copenhagen, Denmark) or placebo (saline). An adult dose of 0.1 ml of resuspended BCG vaccine (intervention) or 0.1 ml of sterile 0.9% NaCl solution (control) is administered intradermally in the upper deltoid area of the right arm. All participants will receive one injection at inclusion, and no further treatment of study participants will take place. Main study endpoint: Days of unplanned absenteeism due to illness within 180 days of randomisation.Secondary study endpoints: The cumulative incidence of documented COVID-19 and the cumulative incidence of hospital admission for any reason within 180 days of randomisation.Randomisation: Randomisation will be done centrally using the REDCap tool with stratification by hospital, sex and age groups (+/- 45 years of age) in random blocks of 4 and 6. The allocation ratio is 1:1.Blinding (masking): Participants will be blinded to treatment. The participant will be asked to leave the room while the allocated treatment is prepared. Once ready for injection, vaccine and placebo will look similar, and the participant will not be able to tell the difference.The physicians administering the treatment are not blinded.Numbers to be randomised (sample size): Sample size: N=1,500. The 1,500 participants will be randomised 1:1 to BCG or placebo with 750 participants in each group.Trial Status: Current protocol version 5.1, from July 6, 2020.Recruitment of study participants started on May 18, 2020 and we anticipate having finished recruiting by the end of December 2020. The trial was registered with EudraCT on April 16, 2020, EudraCT number: 2020-001888-90, and with ClinicalTrials.gov on May 1, 2020, registration number NCT04373291.Full protocol: The full protocol is attached as an additional file, accessible from the Trialswebsite (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


Denture treatment needs of an overdenture population.
Cross-sectional and longitudinal studies have identified that persons wearing overdentures require regular care and maintenance of their dentures and their remaining dentition. This paper reports on a longitudinal study of the denture treatment needs of 284 dentures placed in 254 patients seen over a period of 12 years. At placement of the overdentures, baseline measurements and photographs were made by a single examiner. These measurements included evaluation of the restorations, abutment height above the gingival margin, periodontal probing depths, and oral hygiene. Denture status was evaluated by examining stability, retention, the integrity of the denture bases, and the relationship of the teeth in centric occlusion. Denture hygiene was also assessed. Based upon this examination, the subjects' denture treatment needs were determined. The majority of overdentures were well tolerated, and in general the patients were satisfied with their dentures. The most common denture treatment need was cleaning, followed by adjustment of the denture bases. Relining was a more common problem in the mandibular arch than in the maxillae. The need for occlusal adjustment did not seem to be a problem after the fifth year, but this might have been because of replacement of worn teeth or dentures. The majority of denture repairs were necessitated before the sixth year. Patients who received immediate dentures did not have greater treatment needs. Looseness of the dentures was the primary complaint, followed by sore teeth and denture-bearing tissues. In spite of their complaints, the great majority of the patients were satisfied with their overdentures.


Plectranthus: a review of ethnobotanical uses.
Plectranthus is a large and widespread genus with a diversity of ethnobotanical uses. The genus is plagued with numerous nomenclatural disharmonies that make it difficult to collate accurate data on the uses. The aim of this review is to gather together all ethnobotanical information on Plectranthus and to map the data onto the most up-to-date phylogenetic classification in order to see if there are similar uses among related species and hence provide a framework for the prediction and exploration of new uses of species. The uses of 62 species of Plectranthus were mapped onto a current phylogeny based on DNA sequence data. The phylogeny reveals two major Clades, 1 and 2. The members of Clade 1 (corresponding to the formally recognized genus Coleus) were richer in number and diversity of uses than members of Clade 2 (comprising the remaining species of Plectranthus). The high incidence of synonymy can lead to problems in uncovering a species' ethnobotanical profile. About 30% of all citations of Plectranthus use a synonym and most of the synonyms are attributed to 10 of the most used species, 9 of which are in Clade 1. Members of the 'Coleus' Clade are the most studied group both taxonomically and economically. The higher incidence of study may be as a result of the higher diversity of uses and the fact that species in Clade 1, such as Plectranthus barbatus, Plectranthus amboinicus and Plectranthus mollis, are geographically more widespread than those in Clade 2. Plectranthus species in Clade 1 are frequently used as medicines and are used to treat a range of ailments, particularly digestive, skin, infective and respiratory problems. Plectranthus used as foods, flavours, fodder and materials are also mostly found in Clade 1. Monoterpenoids, sesquiterpenoids, diterpenoids and phenolics have been reported in species of Plectranthus. The abietane diterpenoids are the most diverse of the diterpenoids isolated from species of Plectranthus. The labdane diterpenoid, forskolin, occurs in Plectranthus barbatus and could explain some of the traditional uses of this species. This review highlights the fact that not enough is known about the chemistry of other species of Plectranthus to explain their traditional uses.


Hyperbaric oxygen increases survival following carotid ligation in gerbils.
We studied the effects of graded exposure to hyperbaric (1,875 mm Hg) oxygen therapy in an acute stroke model prepared by unilateral carotid artery interruption in gerbils. Pentobarbital alone, superoxide dismutase alone, two periods of hyperbaric oxygen alone, and each agent combined with hyperbaric oxygen were administered to investigate possible mechanisms of protection from cerebral ischemia. Survival rates and neurologic deficit scores over 5 days in all treated groups were compared with those in a control group. Survival rates in the groups subjected to 2 (63.9 +/- 4.0%) and 4 hours (70.1 +/- 5.2%) of hyperbaric oxygen alone were significantly higher than in the control group (53.6 +/- 4.2%). The group treated with pentobarbital alone also demonstrated increased survival (69.8 +/- 7.0%), but the combination of therapeutic regimens offered no apparent additive protection. By 5 days there were no differences in the neurologic deficit scores of the survivors in the groups. The toxic pulmonary effects of hyperbaric oxygen were assessed in a pilot LD50 study. The pressure used caused no mortality during 4 hours of exposure, and the calculated LD50 was 7.26 hours. This investigation demonstrates that graded doses of hyperbaric oxygen given after the insult increase survival in a gerbil model of stroke.