[Measurement in public health: what pulmonary tuberculosis management has taught us].
Pulmonary tuberculosis rates are increasing worldwide, including in Spain. One of the main challenges when treating this disease is achieving treatment completion, since studies have shown that approximately 30-35% of all patients do not take their medications as intended. The present article explores a continuum of evaluation strategies and performance measures for assessing the effectiveness of community-based programs designed to enhance treatment completion in patients with active pulmonary tuberculosis. Four traditional evaluation strategies (case studies, retrospective and case-control studies, forecasting/modeling, and cost effectiveness analysis) and 2 emerging and promising approaches (quality of life assessment and indicators of the continuum of care) are presented. Several of the evaluation strategies reviewed indicate that treatment programs using directly observed therapy (DOT) that are comprehensive, community-based and patient-centered achieve the highest treatment completion rates. Combinations of these strategies are recommended to create a body of evidence capturing the impact and nuances of community-based public health interventions in improving health outcomes, in this case for patients with pulmonary tuberculosis.


A prospective study analyzing the application of radiofrequency energy and high-voltage, ultrashort pulse duration electrical fields on the quantitative reduction of adipose tissue.
Noninvasive fat reduction is claimed by many device manufacturers, but proof of efficacy has been difficult to establish. This prospective study was designed to measure the reduction of fat thickness and actual volume reduction in 20 female patients treated with an external radiofrequency (RF) device. This device combines RF heat, suction coupled vacuum, and oscillating electrical pulses that induce adipocyte death over time. Patients underwent pre- and post-treatment and intercurrent measurements of weight, body mass index, ultrasonic transcutaneous fat thickness, and 2D and 3D Vectra photography with independent calculation of circumferential and volumetric change. Mean transcutaneous ultrasound thickness at reproducible points was 2.78 cm; at 1-month post-treatment, the mean fat thickness was 1.71 cm. At 3-month post-treatment, the mean fat thickness reduction was 39.6%. Vectra circumference measurements were taken at 10-mm intervals, with postural and breathing cycle control. Independent analysis of serial measurements from + 60 to - 70 mm showed mean abdominal circumference measurement of 2.3 cm. Mean abdominal volume loss was 202.4 and 428.5 cc at 1- and 3-month post-treatment, respectively. Scanning electron microscopy confirmed that permanent cell destruction was caused by irreversible electroporation. Pyroptosis appears to be the mechanism of action.


The uncertainty of crop yield projections is reduced by improved temperature response functions.
Increasing the accuracy of crop productivity estimates is a key element in planning adaptation strategies to ensure global food security under climate change. Process-based crop models are effective means to project climate impact on crop yield, but have large uncertainty in yield simulations. Here, we show that variations in the mathematical functions currently used to simulate temperature responses of physiological processes in 29 wheat models account for >50% of uncertainty in simulated grain yields for mean growing season temperatures from 14 °C to 33 °C. We derived a set of new temperature response functions that when substituted in four wheat models reduced the error in grain yield simulations across seven global sites with different temperature regimes by 19% to 50% (42% average). We anticipate the improved temperature responses to be a key step to improve modelling of crops under rising temperature and climate change, leading to higher skill of crop yield projections.


In silico identification of a core regulatory network of OCT4 in human embryonic stem cells using an integrated approach.
The transcription factor OCT4 is highly expressed in pluripotent embryonic stem cells which are derived from the inner cell mass of mammalian blastocysts. Pluripotency and self renewal are controlled by a transcription regulatory network governed by the transcription factors OCT4, SOX2 and NANOG. Recent studies on reprogramming somatic cells to induced pluripotent stem cells highlight OCT4 as a key regulator of pluripotency. We have carried out an integrated analysis of high-throughput data (ChIP-on-chip and RNAi experiments along with promoter sequence analysis of putative target genes) and identified a core OCT4 regulatory network in human embryonic stem cells consisting of 33 target genes. Enrichment analysis with these target genes revealed that this integrative analysis increases the functional information content by factors of 1.3 - 4.7 compared to the individual studies. In order to identify potential regulatory co-factors of OCT4, we performed a de novo motif analysis. In addition to known validated OCT4 motifs we obtained binding sites similar to motifs recognized by further regulators of pluripotency and development; e.g. the heterodimer of the transcription factors C-MYC and MAX, a prerequisite for C-MYC transcriptional activity that leads to cell growth and proliferation. Our analysis shows how heterogeneous functional information can be integrated in order to reconstruct gene regulatory networks. As a test case we identified a core OCT4-regulated network that is important for the analysis of stem cell characteristics and cellular differentiation. Functional information is largely enriched using different experimental results. The de novo motif discovery identified well-known regulators closely connected to the OCT4 network as well as potential new regulators of pluripotency and differentiation. These results provide the basis for further targeted functional studies.


The role of Hox genes during vertebrate limb development.
The potential role of Hox genes during vertebrate limb development was brought into focus by gene expression analyses in mice (P Dolle, JC Izpisua-Belmonte, H Falkenstein, A Renucci, D Duboule, Nature 1989, 342:767-772), at a time when limb growth and patterning were thought to depend upon two distinct and rather independent systems of coordinates; one for the anterior-to-posterior axis and the other for the proximal-to-distal axis (see D Duboule, P Dolle, EMBO J 1989, 8:1497-1505). Over the past years, the function and regulation of these genes have been addressed using both gain-of-function and loss-of-function approaches in chick and mice. The use of multiple mutations either in cis-configuration in trans-configuration or in cis/trans configurations, has confirmed that Hox genes are essential for proper limb development, where they participate in both the growth and organization of the structures. Even though their molecular mechanisms of action remain somewhat elusive, the results of these extensive genetic analyses confirm that, during the development of the limbs, the various axes cannot be considered in isolation from each other and that a more holistic view of limb development should prevail over a simple cartesian, chess grid-like approach of these complex structures. With this in mind, the functional input of Hox genes during limb growth and development can now be re-assessed.


Arginine-to-lysine substitutions influence recombinant horseradish peroxidase stability and immobilisation effectiveness.
Horseradish Peroxidase (HRP) plays important roles in many biotechnological fields, including diagnostics, biosensors and biocatalysis. Often, it is used in immobilised form. With conventional immobilisation techniques, the enzyme adheres in random orientation: the active site may face the solid phase rather than bulk medium, impeding substrate access and leading to sub-optimal catalytic performance. The ability to immobilise HRP in a directional manner, such that the active site would always face outwards from the insoluble matrix, would maximise the immobilised enzyme's catalytic potential and could increase HRP's range of actual and potential applications. We have replaced arginine residues on the face of glycan-free recombinant HRP opposite to the active site by lysines. Our strategy differs from previous reports of specific HRP immobilisation via an engineered affinity tag or single reactive residue. These conservative Arg-to-Lys substitutions provide a means of multipoint covalent immobilisation such that the active site will always face away from the immobilisation matrix. One triple and one pentuple mutant were generated by substitution of solvent-exposed arginines on the "back" of the polypeptide (R118, R159 and R283) and of residues known to influence stability (K232 and K241). Orientated HRP immobilisation was demonstrated using a modified polyethersulfone (PES) membrane; the protein was forced to orientate its active site away from the membrane and towards the bulk solution phase. Mutant properties and bioinformatic analysis suggested the reversion of K283R to improve stability, thus generating two additional mutants (K118/R159K and R118K/K232N/K241F/R283K). While most mutants were less stable in free solution than wild type rHRP, the quadruple revertant regained some stability over its mutant counterparts. A greater degree of immobilisation on CNBr-activated Sepharosetrade mark was noted with increased lysine content; however, only marginal gains in solvent stability resulted from immobilisation on this latter matrix. Directional, orientated, immobilisation of rHRP mutants onto an activated, modified polyethersulfone membrane has been achieved with excellent retention of catalytic activity; however, re-engineering of acceptable stability characteristics into the "immobilisation mutants" will determine their applicability in diagnosis and biosensor development.


Overexpression of monocarboxylate transporter-1 (SLC16A1) in mouse pancreatic β-cells leads to relative hyperinsulinism during exercise.
Exercise-induced hyperinsulinism (EIHI) is an autosomal dominant disorder characterized by inappropriate insulin secretion in response to vigorous physical exercise or pyruvate injection. Activating mutations in the monocarboxylate transporter-1 (MCT1, SLC16A1) promoter have been linked to EIHI. Expression of this pyruvate transporter is specifically repressed (disallowed) in pancreatic β-cells, despite nearly universal expression across other tissues. It has been impossible to determine, however, whether EIHI mutations cause MCT1 expression in patient β-cells. The hypothesis that MCT1 expression in β-cells is sufficient to cause EIHI by allowing entry of pyruvate and triggering insulin secretion thus remains unproven. Therefore, we generated a transgenic mouse capable of doxycycline-induced, β-cell-specific overexpression of MCT1 to test this model directly. MCT1 expression caused isolated islets to secrete insulin in response to pyruvate, without affecting glucose-stimulated insulin secretion. In vivo, transgene induction lowered fasting blood glucose, mimicking EIHI. Pyruvate challenge stimulated increased plasma insulin and smaller excursions in blood glucose in transgenic mice. Finally, in response to exercise, transgene induction prevented the normal inhibition of insulin secretion. Forced overexpression of MCT1 in β-cells thus replicates the key features of EIHI and highlights the importance of this transporter's absence from these cells for the normal control of insulin secretion.


Shared medical appointments for Aboriginal and Torres Strait Islander men.
Aboriginal and Torres Strait Islander health is generally the worst of any population group in Australia. Inaccessibility to health services is one possible cause of this. Shared medical appointments (SMAs) appear to be a culturally competent and appropriate way of improving access to, and the quality of, primary healthcare services for Aboriginal and Torres Strait Islander peoples. The objective of this article is to assess the acceptability and appropriateness of SMAs as an adjunct process in primary care for Aboriginal and Torres Strait Islander men. As part of a broader study on SMAs, three SMA sessions were delivered at an Aboriginal men's health centre in northern New South Wales. One-day training sessions in SMA facilitation were also provided to two groups of 12-14 Aboriginal health workers (AHWs). Mixed methods were used to assess patient and provider satisfaction, subjective outcomes, and operational procedures in the SMA groups, as well as interest in the SMA process by AHWs. Satisfaction with SMAs among Aboriginal men was unanimously positive, with the numbers in the group increasing over time. Patients most enjoyed the 'yarn up' nature of SMAs with peer support, which reduced the 'scary' and culturally 'unnatural' nature of one-on-one consultations with a general practitioner (GP). AHWs who were trained to a level to conduct SMAs saw this as an effective way of improving cultural competence in, and accessibility of, their various Aboriginal health services. The results, though not generalisable, suggest that SMAs may offer a culturally safe and appropriate tool to enhance Aboriginal and Torres Strait Islander peoples' access to primary care.


Cost savings at the end of life. What do the data show?
Medical care at the end of life consumes 10% to 12% of the total health care budget and 27% of the Medicare budget. Many people claim that increased use of hospice and advance directives and lower use of high-technology interventions for terminally ill patients will produce significant cost savings. However, the studies on cost savings from hospice and advance directives are not definitive. The 3 randomized trials show no savings from these interventions, but either they are too small for confidence in their negative results or their intervention and cost accounting are flawed. The nonrandomized trials of hospice and advance directives show a wide range of savings, from 68% to none. Five methodological issues obscure the assessment of these studies: (1) selection bias in those patients who use hospice and advance directives, (2) the different time frames of assessing the costs, (3) the limited types of medical costs evaluated, (4) the variability of reporting the savings, and (5) the lack of generalizability of the findings to other patient populations. A more definitive study that assessed patients' end-of-life care preferences, use of hospice and advance directives, and direct and indirect costs would be desirable. In the absence of such a study, the existing data suggest that hospice and advance directives can save between 25% and 40% of health care costs during the last month of life, with savings decreasing to 10% to 17% over the last 6 months of life and decreasing further to 0% to 10% over the last 12 months of life. These savings are less than most people anticipate. Nevertheless, they do indicate that hospice and advance directives should be encouraged because they certainly do not cost more and they provide a means for patients to exercise their autonomy over end-of-life decisions.


High affinity antibodies to Plasmodium falciparum merozoite antigens are associated with protection from malaria.
Malaria kills almost 1 million people every year, but the mechanisms behind protective immunity against the disease are still largely unknown. In this study, surface plasmon resonance technology was used to evaluate the affinity (measured as k(d)) of naturally acquired antibodies to the Plasmodium falciparum antigens MSP2 and AMA1. Antibodies in serum samples from residents in endemic areas bound with higher affinities to AMA1 than to MSP2, and with higher affinities to the 3D7 allele of MSP2-3D7 than to the FC27 allele. The affinities against AMA1 and MSP2-3D7 increased with age, and were usually within similar range as the affinities for the monoclonal antibodies also examined in this study. The finding of MSP2-3D7 type parasites in the blood was associated with a tendency for higher affinity antibodies to both forms of MSP2 and AMA1, but this was significant only when analyzing antibodies against MSP2-FC27, and individuals infected with both allelic forms of MSP2 at the same time showed the highest affinities. Individuals with the highest antibody affinities for MSP2-3D7 at baseline had a prolonged time to clinical malaria during 40 weeks of follow-up, and among individuals who were parasite positive at baseline higher antibody affinities to all antigens were seen in the individuals that did not experience febrile malaria during follow up. This study contributes important information for understanding how immunity against malaria arises. The findings suggest that antibody affinity plays an important role in protection against disease, and differs between antigens. In light of this information, antibody affinity measurements would be a key assessment in future evaluation of malaria vaccine formulations.


Effect of amino acid substitutions on the catalytic and regulatory properties of aspartate transcarbamoylase.
Although intensive investigations have been conducted on the allosteric enzyme, aspartate transcarbamoylase, which catalyzes the first committed reaction in the biosynthesis of pyrimidines in Escherichia coli, little is known about the role of individual amino acid residues in catalysis or regulation. Two inactive enzymes produced by random mutagenesis have been characterized previously but the loss of activity is probably attributable to changes in the folding of the chains stemming from the introduction of charged and bulky residues (Asp for Gly-128 and Phe for Ser-52). Site-directed mutagenesis of pyrB, which encodes the catalytic chains of the enzyme, was used to probe the functional roles of several amino acids by making more conservative substitutions. Replacement of Lys-84 by either Gln or Arg leads to virtually inactive enzymes, confirming chemical studies indicating that Lys-84 is essential for catalysis. In contrast, substitution of Gln for Lys-83 has only a slight effect on enzyme activity, whereas chemical modification causes considerable inactivation. Gln-133, which has been shown by x-ray crystallography to reside near the contact region between the catalytic and regulatory chains, was replaced by Ala. This substitution has little effect on catalytic activity but leads to a marked increase in cooperativity. The Gln-83 mutant, in contrast, exhibits much less cooperativity. Since a histidine residue may be involved in catalysis and His-134 has been shown by x-ray diffraction studies to be in close proximity to the site of binding of a bisubstrate analog, His-134 was replaced by Ala, yielding a mutant with only 5% wild-type activity, considerable cooperativity, and lower affinity for aspartate and carbamoylphosphate. All of the mutants, unlike those in which charged or bulky residues replaced small side chains, bind the bisubstrate analog, which promotes the characteristic "swelling" of the enzymes indicative of the allosteric transition.


Displaced femoral neck fatigue fractures in military recruits.
Displaced fatigue fractures of the femoral neck are uncommon, but they can lead to substantial patient morbidity. This study was performed to examine the incidence, long-term consequences, radiographic findings, risk factors, and complications associated with this fracture. Between 1975 and 1994, twenty-one military recruits sustained a displaced fatigue fracture of the femoral neck. Nineteen patients were followed for an average of eighteen years. Data regarding the population at risk, hospital records, initial and follow-up radiographs, and physical findings were analyzed. The impact of instructions from the Finnish Defense Forces, Department of Medical Services, provided in 1986 for prevention of femoral neck fatigue fractures was assessed. At our institution, the incidence of displaced fatigue fractures of the femoral neck was 5.3/100,000 service years from 1975 to 1986, prior to the introduction of the prevention regimen in 1986, and it was 2.3/100,000 service years (95% confidence interval, 0.11 to 1.31) from 1987 to 1994. The rate of Garden type-IV fractures decreased from 3.8 to 0/100,000 service years (95% confidence interval, 0 to 0.66) between the first and second time-periods. The detection of nondisplaced symptomatic fatigue fractures of the femoral neck increased from 15.5 to 53.2/100,000 service years (95% confidence interval, 2.27 to 5.21) between the two time-periods. Eighteen of the nineteen patients had had prodromal symptoms prior to the fracture displacement. Following fracture treatment, six patients had delayed union or nonunion of the fracture. Osteonecrosis of the femoral head developed in six patients and was significantly associated (p = 0.001) with shortening of the femoral neck. Severe osteoarthritis developed in eight patients. A displaced fatigue fracture of the femoral neck leads to long-term morbidity in a high percentage of patients. Most patients have prodromal symptoms, which provide an opportunity to prevent fracture displacement. Our results indicate that, in a military setting, an educational program can diminish the incidence of fatigue fracture displacement by increasing the awareness of these fractures and their prodromal symptoms and by facilitating diagnosis in the early stages before displacement occurs. Therapeutic Level III.


AMPA receptors do the electric slide.
How the synapse is organized and how its organization changes during events that result in long-term changes in synaptic efficacy is the subject of intense study. Various anchoring proteins work in concert to organize the postsynaptic side of the membrane, and the interactions of these proteins can be altered by synaptic activity. DeSouza and Ziff discuss the evidence that the reversible palmitoylation of the postsynaptic density protein PSD-95 may result in the movement of AMPA-type glutamate receptors into and out of lipid raft domains, ultimately controlling AMPA receptor accumulation at the postsynaptic membrane.


Formal and Informal Care of Community-Living Older People: A Population-Based Study from the Swedish National Study on Aging and Care.
Study formal and informal care of community-living older people in the Swedish National study of Aging and Care (SNAC). Cross-sectional, population based cohort. Three areas in Sweden: Municipality of Nordanstig, Stockholm and Skåne County. 3,338 persons ≥72 years. Patterns and amounts of informal and formal care by cognition and area of residence. 73% received no care; 14% formal care; and 17% informal care (7% received both). In the whole study population, including those who used no care, individuals in small municipalities received 9.6 hours of informal care/month; in mid-size municipalities, 6.6; and in urban areas, 5.6. Users of informal care received 33.1 hours of informal care/month in small municipalities, 54.6 in mid-size municipalities and 36.1 in urban areas. Individuals with cognitive impairment received 14.1 hours of informal care/month, 2.7 times more than people with no/slight impairment. In the whole study population, individuals in small municipalities received an average of 3.2 hours of formal care/month; in mid-size municipalities 1.4; and in urban areas, 2.6. Corresponding figures for formal care users were 29.4 hours in small municipalities, 13.6 in mid-size municipalities and 16.7 in urban areas. Formal care users received 7.1 hours, and informal care users, 5.9 hours for each hour/month received by people in the study population as a whole. More informal than formal care was provided. Informal care is more frequent in small municipalities than urban areas and for those with than without cognitive impairment. The relationship between data on the whole population and the data on users or care indicates that population-based data are needed to avoid overestimates of care.


Thirteen years' experience of 893 PGD cycles for monogenic disorders in a publicly funded, nationally regulated regional hospital service.
This study provides an overview of preimplantation genetic diagnosis (PGD) for single gene diseases and the management of expanding indications in the context of a fully financially covered service at Montpellier's regional hospital centre. Within the framework of a restrictive law ruling PGD in France, only the parental genetic risk can be studied in embryos (concurrent aneuploidy screening is not allowed). PCR-based techniques were developed combining mutation detection and closely linked short tandem repeat markers within or flanking the affected genes, and set up more than 100 different robust fluorescent multiplex assays for 61 monogenic disorders. This strategy was used to analyse blastomeres from cleavage-stage embryos. Overall, 893 cycles were initiated in 384 couples; 727 cycles proceeded to oocyte retrieval and 608 cycles to embryo transfer, resulting in 184 deliveries. Clinical pregnancy rate per transfer, implantation and miscarriage rates were 33.6%, 25.1% and 8.8%, respectively. Our PGD programme resulted in the birth of 214 healthy babies for 162 out of 358 couples (45.3%), constituting a relevant achievement within an organizational framework that does not allow aneuploidy screening but provides equal access to PGD, both geographically and socioeconomically. This is a rare example of a fully free-of-charge PGD service.


Health differentials between citizens and immigrants in Europe: A heterogeneous convergence.
The literature on immigration and health has provided mixed evidence on the health differentials between immigrants and citizens, while a growing body of evidence alludes to the unhealthy assimilation of immigrants. Relying on five different health measures, the present paper investigates the heterogeneity in health patterns between immigrants and citizens, and also between immigrants depending on their country of origin. We use panel data on more than 100,000 older adults living in nineteen European countries. Our panel data methodology allows for unobserved heterogeneity. We document the existence of a healthy immigrant effect, of an unhealthy convergence, and of a reversal of the health differentials between citizens and immigrants over time. We are able to estimate the time threshold after which immigrants' health becomes worse than that of citizens. We further document some heterogeneity in the convergence of health differentials between immigrants and citizens in Europe. Namely, the unhealthy convergence is more pronounced in terms of chronic conditions for immigrants from low-HDI countries, and in terms of self-assessed health and body-mass index for immigrants from medium- and high-HDI countries.


A Wireless Sensor for Real-Time Monitoring of Tensile Force on Sutured Wound Sites.
A new wireless sensor was designed, fabricated, and applied for in situ monitoring of tensile force at a wound site. The sensor was comprised of a thin strip of magnetoelastic material with its two ends connected to suture threads for securing the sensor across a wound repair site. Since the sensor was remotely interrogated by applying an ac magnetic field and capturing the resulting magnetic field, it did not require direct wire connections to an external device or internal battery for long-term use. Due to its magnetoelastic property, the application of a tensile force changed the magnetic permeability of the sensor, altering the amplitude of the measured magnetic field. This study presents two sensor designs: one for high and one for low-force ranges. A sensor was fabricated by directly adhering the magnetoelastic strip to the suture. This sensor showed good sensitivity at low force, but its response saturated at about 1.5 N. To monitor high tensile force, the magnetoelastic strip was attached to a metal strip for load sharing. The suture thread was attached to the both ends of the metal strip so only a fraction of the applied force was directed to the sensor, allowing it to exhibit good sensitivity even at 44.5 N. The sensor was applied to two ex vivo models: a sutured section of porcine skin and a whitetail deer Achilles tendon. The results demonstrate the potential for in vivo force monitoring at a wound repair site.


Changes in the immune system in rodent models of depression.
This review summarizes some of the evidence which implicates an increase in the peripheral and central secretion of pro-inflammatory cytokines in the behavioural changes seen in some stress-induced and brain lesion models of depression. Following a consideration of the role of cytokines in the periphery and the brain, evidence is presented suggesting that pro-inflammatory cytokines alter the function of monoamine neurotransmitters which have been implicated in severe stress and in major depression. These changes occur in the presence of elevated glucocorticoid concentrations which suggests that immune activation is correlated with a decrease in the sensitivity of the glucocorticoid receptors on immune cells in addition to those occurring in the brain. The review concludes with a brief account of the various rodent models of depression in which evidence of immune activation as been demonstrated.


Saccharomyces cerevisiae Med9 comprises two functionally distinct domains that play different roles in transcriptional regulation.
Mediator is one of the most important co-activators that function in eukaryotic transcriptional regulation. In Saccharomyces cerevisiae, Mediator is comprised of 25 subunits belonging to four structurally distinct modules: Head, Middle, Tail, and Cyc-C. Although each module plays a critical role in the regulation of a distinct set of genes, the precise molecular mechanisms remain unclear. To gain new insight into the role of the less-characterized Middle module, we analyzed the function of Med9 by constructing a set of mutants and subjecting them to a range of in vivo and in vitro assays. Our results demonstrate that Med9 has two functional domains. The species-specific amino-terminal half (aa 1-63) plays a regulatory role in transcriptional regulation in vivo and in vitro. In contrast, the well-conserved carboxy-terminal half (aa 64-149) has a more fundamental function involved in direct binding to the amino-terminal portions of Med4 and Med7 and the assembly of Med9 into the Middle module. Importantly, activator-dependent recruitment of TBP and Taf11 to the promoter is differentially affected in med9 extracts and in extracts lacking Mediator. Add-back experiments indicate that some unidentified factor(s) in med9 extracts may impact the binding of TFIID to the promoter.


Distance from a distance: psychological distance reduces sensitivity to any further psychological distance.
What is the difference between far and further? Investigations into such psychological distancing--removal from an egocentric reference point--have suggested similarities between geographical space, time, probability, and social distance. We draw on these similarities to propose that experiencing any kind of distance will reduce sensitivity to any other distance. Nine studies varied the initial distance of an event and assessed sensitivity to a second distance. Consistently, people were less responsive to a given span of distance when it was distal versus proximal. This effect held using each of the four distances as the initial instantiation of distance; it also held using each dimension to assess sensitivity to distance (i.e., as the second distance dimension). These findings suggest that the dimensions of psychological distance share a common, interchangeable meaning and that the cross-dimension difference between far and further is less than that between near and far.


Dicer-labile PEG conjugates for siRNA delivery.
Poly(ethylene glycol) (PEG) conjugates of Dicer-substrate small interfering RNA (DsiRNA) have been prepared to investigate a new siRNA release strategy. 3'-sense or 5'-antisense thiol-modified, blunt-ended DsiRNAs, inhibiting enhanced green fluorescent protein (eGFP) expression, were covalently conjugated to PEG with varying molecular weights (2, 10, and 20 kg/mol) through a stable thioether bond using a Michael addition reaction. The DsiRNA conjugates with 2 kg/mol PEG (both 3'-sense or 5'-antisense strand conjugated) and the 10 kg/mol PEG conjugated to the 3'-sense strand of DsiRNA were efficiently cleaved by recombinant human Dicer to 21-mer siRNA, as determined by gel electrophoresis. Importantly, 2 and 10 kg/mol PEG conjugated to the 3'-sense strand of DsiRNA showed potent gene silencing activity in human neuroblastoma (SH-EP) cells, stably expressing eGFP, at both the mRNA and protein levels. Moreover, the 10 kg/mol PEG conjugates of the 3'-sense strand of DsiRNA were less immunogenic when compared with the unmodified DsiRNA, determined via an immune stimulation assay on human peripheral blood mononuclear cells.


Positive Low Cloud and Dust Feedbacks Amplify Tropical North Atlantic Multidecadal Oscillation.
The Atlantic Multidecadal Oscillation (AMO) is characterized by a horseshoe pattern of sea surface temperature (SST) anomalies and has a wide range of climatic impacts. While the tropical arm of AMO is responsible for many of these impacts, it is either too weak or completely absent in many climate model simulations. Here we show, using both observational and model evidence, that the radiative effect of positive low cloud and dust feedbacks is strong enough to generate the tropical arm of AMO, with the low cloud feedback more dominant. The feedbacks can be understood in a consistent dynamical framework: weakened tropical trade wind speed in response to a warm middle latitude SST anomaly reduces dust loading and low cloud fraction over the tropical Atlantic, which warms the tropical North Atlantic SST. Together they contribute to appearance of the tropical arm of AMO. Most current climate models miss both the critical wind speed response and two positive feedbacks though realistic simulations of them may be essential for many climatic studies related to the AMO.


Gas fermentation: cellular engineering possibilities and scale up.
Low carbon fuels and chemicals can be sourced from renewable materials such as biomass or from industrial and municipal waste streams. Gasification of these materials allows all of the carbon to become available for product generation, a clear advantage over partial biomass conversion into fermentable sugars. Gasification results into a synthesis stream (syngas) containing carbon monoxide (CO), carbon dioxide (CO2), hydrogen (H2) and nitrogen (N2). Autotrophy-the ability to fix carbon such as CO2 is present in all domains of life but photosynthesis alone is not keeping up with anthropogenic CO2 output. One strategy is to curtail the gaseous atmospheric release by developing waste and syngas conversion technologies. Historically microorganisms have contributed to major, albeit slow, atmospheric composition changes. The current status and future potential of anaerobic gas-fermenting bacteria with special focus on acetogens are the focus of this review.


Cytokine-stimulated chemotaxis of human neutrophils in a 3-D conjoined fibrin gel assay.
The ability of neutrophils to migrate through three-dimensional (3-D) tissues in response to chemical stimuli is critical to their host defense function. However, studies characterizing stimulated migration in vitro have been largely limited to two-dimensional (2-D) surfaces. In this study, we have employed direct observation methods to quantify human neutrophil migration in 3-D fibrin gel using time-lapse video microscopy and automated cell tracking methods. A novel 3-D conjoined gel assay was developed to establish experimentally quantifiable and theoretically predictable diffusion gradients of chemotactic factors. This assay was used to measure objective migration parameters, namely the random motility and chemotaxis coefficients, in response to the cytokine, interleukin-8 (IL-8). The random motility coefficient, mu, showed a biphasic dependence on IL-8 concentration with a maximum of 1.1 x 10(-8) cm2/s at 5 x 10(-8) M IL-8; no significant motility was observed in the absence of IL-8. We further established the dependence of cell orientation bias, phi, on the concentration and gradient steepness (i.e., specific gradient, SG) of IL-8. Results indicate that phi increases with increasing SG, provided the concentration is maintained sufficiently low, which we conjecture to result from minimizing IL-8 receptor down-regulation. The chemotaxis coefficient, chi, was maximum at an intermediate SG for both IL-8 concentrations studied. We also examined the applicability of this assay to estimate mu and chi from indirect measurements of chemotaxis, namely the simpler measurement of cell redistribution after a prescribed incubation time, as opposed to direct cell tracking measurements. By virtue of measuring chi, this is the first quantitatively objective study of mammalian cell chemotaxis in a physiologically relevant 3-D gel and, in particular, of neutrophil chemotaxis on any substratum in response to the physiologically relevant chemotactic factor, IL-8.


What makes one feel eustress or distress in quarantine? An analysis from conservation of resources (COR) theory.
Health quarantines produce serious deterioration in psychological health, which becomes more affected the longer the quarantine lasts. According to the Conservation of Resources theory from Hobfoll (1989, American Psychologist, 44, 513), those people who have a good supply of resources will be able to cope better with the adversities and will show less distress. The objective of this research is to identify what are the resources that, in a situation of confinement under the threat of COVID-19, predict eustress or well-being, and the loss or lack of which resources predict distress or discomfort. A total of 839 people complete an online questionnaire during the first week of COVID-19 confinement in Spain. The sample is weighted to obtain a distribution that is similar to the Spanish population. Using multiple linear regression analysis, factors are identified that are associated with eustress and distress based on the Conservation of Resources theory. A model is identified that explains 55% of the variance of eustress consisting mostly of personal resources, with vitality as the recourse having the most weight. The factors that explain distress (18.9% of the variance) are those related to work (employment situation, work satisfaction, and time devoted to work) and conditions in the home (space). The models that predict eustress and distress are completely different. Based on these results, a series of recommendations are proposed aimed at increasing eustress and reducing distress in a situation of confinement. Additionally, proposals are offered for future research.


Misclassification and discordance of measured blood pressure from patient's true blood pressure in current clinical practice: a clinical trial simulation case study.
Treatment decisions for hypertension using sphygmomanometer based measurements and the current clinical practice paradigm do not account for the timing of blood pressure (BP) measurement. This study aimed to evaluate the clinical implications of discordance between measured and true BP, to quantify BP misclassification rate at a typical clinical visit in current clinical practice, and to propose a BP calibration system to decrease the impact of timing of BP measurement. A clinical trial simulation case study was performed using an in silico Monte Carlo Simulation approach. The time-courses of BPs with and without an antihypertensive treatment effect were simulated from a baseline BP model combined with an antihypertensive PK/PD model. Virtual subject characteristics were sampled from the FDA internal database. The baseline BP model was qualified using observed 24 h ambulatory BP monitoring (ABPM) data from 225 subjects by a visual predictive check as well as a global sensitivity analysis. First of all, our results showed that the measured cuff BP in current typical clinical practice deviated from the true values. (1) Cuff BP deviated from the true values by more than 5 mmHg in 57 % (95 % CI: 54-61 %) of patients and more than 10 mmHg in 26 % (95 % CI: 22-32 %) of patients respectively. (2) These discordances were reduced to 28 % (deviation ≥ 5 mmHg, 95 % CI: 18-40 %) and 9 % (deviation ≥ 10 mmHg, 95 % CI: 4-18%) of patients assuming perfect sphygmomanometer measurement and thus represent the contribution of ignoring the daily circadian rhythm of BP. Secondly, our results showed 23-32 % of patients were misclassified to an incorrect BP category for a casual clinical visit based on JNC 7 guideline. In addition, the accuracy of the measured cuff BP varied by time of clinic visit. Specifically, 11:00 AM to 3:00 PM was identified to be the better time frame, while times before 9:00 AM were the worst time frame. Therefore, clinic visit time may need to be adjusted accordingly. Finally, we proposed an easy BP calibration method for clinic use to adjust for time of day differences due to circadian variability in case that the desirable clinic visit time cannot be tailored for practical reasons.


Indications and outcomes for revision of gold weight implants in upper eyelid loading.
Gold weights are effective for upper eyelid loading in patients with lagophthalmos. Complications include poor cosmesis, migration, extrusion, allergy and astigmatism. The authors looked at indications for revision of primary gold weights inserted using a high pretarsal placement and outcomes following correction. A retrospective review of 107 consecutive primary gold weight implants in 95 patients with lagophthalmos in a single centre over a 5-year period. Implant placement utilised a combined high pretarsal placement, levator recession and fixation. Revision surgery included repositioning, removal or exchange. Blinded assessment of eyelid parameters, including cosmesis, was performed by an independent reviewer using photographs from each revision case taken preoperatively and 6 months postoperatively. Mean follow-up 2.5 years (range 1-5) with 15/107 (14%) eyelids revised, the majority within 12 months of the primary procedure. Five eyelids required up to 3 further revisions, giving 21 revisions in total. Indications included prominent implants in 15/21 (71%) revisions; poor eyelid contour in 14/21 (67%, 9 drooped and 5 flattened eyelids); extrusion in 2/21 (10%); persistent erythema in 8/21 (29%, 5 gold allergies and 1 extrusion). Revisions consisted of platinum chain exchange (6), replacement (3), repositioning (8) and removal (4). Following final revision, eyelid contour returned to normal and five eyelids demonstrated mild prominence. High pretarsal placement was successful in treating lagophthalmos, with a complication rate of 1 in 6 requiring a revision procedure, the majority within 12 months. Main indications were unsatisfactory cosmesis from prominence of implant and poor eyelid contour.


Use of Sodium-Glucose Cotransporter 2 Inhibitors in Patients With Type 1 Diabetes and Rates of Diabetic Ketoacidosis.
To estimate real-world off-label use of sodium-glucose cotransporter 2 (SGLT2) inhibitors in patients with type 1 diabetes, estimate rates of diabetic ketoacidosis (DKA), and compare them with DKA rates observed in sotagliflozin clinical trials. We identified initiators of SGLT2 inhibitors in the Sentinel System from March 2013 to June 2018, determined the prevalence of type 1 diabetes using a narrow and a broad definition, and measured rates of DKA using administrative claims data. Standardized incidence ratios (SIRs) were calculated using age- and sex-specific follow-up time in Sentinel and age- and sex-specific DKA rates from sotagliflozin trials 309, 310, and 312. Among 475,527 initiators of SGLT2 inhibitors, 0.50% and 0.92% met narrow and broad criteria for type 1 diabetes, respectively. Rates of DKA in the narrow and broad groups were 7.3/100 person-years and 4.5/100 person-years, respectively. Among patients who met narrow criteria for type 1 diabetes, rates of DKA were highest for patients aged 25-44 years, especially females aged 25-44 years (19.7/100 person-years). More DKA events were observed during off-label use of SGLT2 inhibitors in Sentinel than would be expected based on sotagliflozin clinical trials (SIR = 1.83; 95% CI 1.45-2.28). Real-world off-label use of SGLT2 inhibitors among patients with type 1 diabetes accounted for a small proportion of overall SGLT2 inhibitor use. However, the risk for DKA during off-label use was notable, especially among young, female patients. Although real-word rates of DKA exceeded the expectation based on clinical trials, results should be interpreted with caution due to differences in study methods, patient samples, and study drugs.


MHC control of CD4+ T cell subset activation.
The present results demonstrate that CD4+ T cells activated in the primary in vivo response to antigen produce distinct patterns of cytokines depending upon the MHC class II haplotype of the responding mice. I-As mice were found to selectively activate IL-2/IFN-gamma-producing CD4+ T cells, whereas I-Ab mice exhibited selective activation of IL-4-producing CD4+ T cells in response to collagen IV. The effector response phenotype was found to correlate with the cytokine phenotype of CD4+ T cells activated in vivo; IL-2/IFN-gamma-producing cells giving rise to proliferative (cell-mediated) responses, IL-4-producing cells leading to secondary IgG (humoral) responses. Together the data support the notion that the outcome of a given immune response (e.g., protection vs. onset, tolerance vs. autoimmunity) may be determined in part by the type of CD4+ T cells initially activated by antigen. Moreover, the present experiments demonstrate for the first time that polymorphism in class II MHC can determine such selective activation of different cytokine-producing CD4+ T cell phenotypes.


Use of an endoscope-compatible probe to detect colonic dysplasia with Fourier transform infrared spectroscopy.
Fourier transform infrared (FTIR) spectroscopy is sensitive to the molecular composition of tissue and has the potential to identify premalignant tissue (dysplasia) as an adjunct to endoscopy. We demonstrate collection of mid-infrared absorption spectra with a silver halide (AgCl(0.4)Br(0.6)) optical fiber and use spectral preprocessing to identify optimal subranges that classify colonic mucosa as normal, hyperplasia, or dysplasia. We collected spectra (n=83) in the 950 to 1800 cm(-1) regime on biopsy specimens obtained from human subjects (n=37). Subtle differences in the magnitude of the absorbance peaks at specific wave numbers were observed. The best double binary algorithm for distinguishing normal-versus-dysplasia and hyperplasia-versus-dysplasia was determined from an exhaustive search of spectral intervals and preprocessing techniques. Partial least squares discriminant analysis was used to classify the spectra using a leave-one-subject-out cross-validation strategy. The results were compared with histology reviewed independently by two gastrointestinal pathologists. The optimal thresholds identified resulted in an overall sensitivity, specificity, accuracy, and positive predictive value of 96%, 92%, 93%, and 82%, respectively. These results indicated that mid-infrared absorption spectra collected remotely with an optical fiber can be used to identify colonic dysplasia with high accuracy, suggesting that continued development of this technique for the early detection of cancer is promising.


Treatment of ulcerative reflux oesophagitis with colloidal bismuth subcitrate in combination with cimetidine.
Twenty patients took part in a controlled double blind study comparing the efficacy of colloidal bismuth and cimetidine (800 mg at night) with cimetidine alone in the treatment of ulcerative reflux oesophagitis. Colloidal bismuth 120 mg was administered through an intraoesophageal tube four times a day. Cimetidine with colloidal bismuth gives significantly (p less than 0.001) better results than cimetidine alone. Of 10 patients treated with cimetidine and bismuth, seven had no endoscopic signs of oesophagitis after three weeks and three had grade I oesophagitis. Of 10 patients treated with cimetidine alone, one had grade I oesophagitis and three grade II oesophagitis after three weeks. The remaining six still had grade III oesophagitis. This study also shows that in nine of 10 patients reflux oesophagitis was accompanied by infection with campylobacter like organisms (CLO). After treatment the bacteria disappeared promptly in five patients receiving combination therapy and in two of four treated with cimetidine alone. The possible role of CLO in reflux oesophagitis is uncertain.


Characterization and sequence comparison of temperature-regulated chaperonins from the hyperthermophilic archaeon Archaeoglobus fulgidus.
We have cloned and sequenced the genes encoding two chaperonin subunits (Cpn-alpha and Cpn-beta), from Archaeoglobus fulgidus, a sulfate-reducing hyperthermophilic archaeon. The genes encode proteins of 545 amino acids with calculated Mr of 58 977 and 59 683. Both proteins have been identified in cytoplasmic fractions of A. fulgidus by Western analysis using antibodies raised against one of the subunits expressed in Escherichia coli, and by N-terminal amino acid sequencing of chaperonin complexes purified by immunoprecipitation. The chaperonin genes appear to be under heat shock regulation, as both proteins accumulate following temperature shift-up of growing A. fulgidus cells, implying a role of the chaperonin in thermoadaptation. Canonical Box A and Box B archaeal promoter sequences, as well as additional conserved putative signal sequences, are located upstream of the start codons. A phylogenetic analysis using all the available archaeal chaperonin sequences, suggests that the alpha and beta subunits are the results of late gene duplications that took place well after the establishment of the main archaeal evolutionary lines.


Depression, anxiety, and resting frontal EEG asymmetry: a meta-analytic review.
Emotion-related disturbances, such as depression and anxiety, have been linked to relative right-sided resting frontal electroencephalograph (EEG) asymmetry among adults and infants of afflicted mothers. However, a somewhat inconsistent pattern of findings has emerged. A meta-analysis was undertaken to (a) evaluate the magnitude of effects across EEG studies of resting frontal asymmetry and depression, anxiety, and comorbid depression and anxiety and (b) determine whether certain moderator variables could help reconcile inconsistent findings. Moderate effects of similar magnitude were obtained for the depression and anxiety studies, whereas a smaller effect emerged for comorbid studies. Three moderating variables predicted effect sizes: (a) Shorter EEG recording periods were associated with larger effects among adults, (b) different operationalizations of depression yielded effects of marginally different magnitudes, and (c) younger infant samples showed larger effects than older ones. The current data support a link between resting frontal EEG asymmetry and depression and anxiety and provide a partial account of inconsistent findings across studies.


A monoclonal antibody that recognizes a potential coeliac-toxic repetitive pentapeptide epitope in gliadins.
Antibodies that detect coeliac-toxic prolamins from wheat, barley and rye are important tools for controlling the diet of coeliac disease patients. Recently, a monoclonal antibody R5 that recognizes wheat gliadin, barley hordein and rye secalin equally was described. In this study, the epitope recognized by R5 was investigated. Both a phage-displayed heptapeptide library and overlapping peptides spanning the sequence of alpha- and gamma-type gliadins (pepscan) were screened for binding of R5. Both techniques yielded comparable pentapeptide consensus sequences (phage display QXPW/FP; pepscan QQPFP). According to recent observations, this peptide stretch may be of key importance in the pathogenicity of coeliac disease. This sequence occurs repetitively in prolamins (in gamma- and omega-type prolamins more frequently than in alpha-type prolamins) together with several homologous peptide stretches, which are recognized less strongly. R5 seems to be a good candidate for the specific detection of putative coeliac disease-active sequences in prolamins and thus represents a valuable tool for the quality control of gluten-free food.


Multiparticle adhesive dynamics. Interactions between stably rolling cells.
A novel numerical simulation of adhesive particles (cells) reversibly interacting with an adhesive surface under flow is presented. Particle--particle and particle--wall hydrodynamic interactions in low Reynolds number Couette flow are calculated using a boundary element method that solves an integral representation of the Stokes equation. Molecular bonds between surfaces are modeled as linear springs and stochastically formed and broken according to postulated descriptions of force-dependent kinetics. The resulting simulation, Multiparticle Adhesive Dynamics, is applied to the problem of selectin-mediated rolling of hard spheres coated with leukocyte adhesion molecules (cell-free system). Simulation results are compared to flow chamber experiments performed with carbohydrate-coated spherical beads rolling on P-selectin. Good agreement is found between theory and experiment, with the main observation being a decrease in rolling velocity with increasing concentration of rolling cells or increasing proximity between rolling cells. Pause times are found to increase and deviation motion is found to decrease as pairs of rolling cells become closer together or align with the flow.


Capuchin monkeys display affiliation toward humans who imitate them.
During social interactions, humans often unconsciously and unintentionally imitate the behaviors of others, which increases rapport, liking, and empathy between interaction partners. This effect is thought to be an evolutionary adaptation that facilitates group living and may be shared with other primate species. Here, we show that capuchin monkeys, a highly social primate species, prefer human imitators over non-imitators in a variety of ways: The monkeys look longer at imitators, spend more time in proximity to imitators, and choose to interact more frequently with imitators in a token exchange task. These results demonstrate that imitation can promote affiliation in nonhuman primates. Behavior matching that leads to prosocial behaviors toward others may have been one of the mechanisms at the basis of altruistic behavioral tendencies in capuchins and in other primates, including humans.


Resolution of the polarisation of ideologies and approaches in psychiatry.
The uniqueness of Psychiatry as a medical speciality lies in the fact that aside from tackling what it considers as illnesses, it has perchance to comment on and tackle many issues of social relevance as well. Whether this is advisable or not is another matter; but such a process is inevitable due to the inherent nature of the branch and the problems it deals with. Moreover this is at the root of the polarization of psychiatry into opposing psychosocial and biological schools. This gets reflected in their visualization of scope, in definitions and in methodology as well. Whilst healthy criticism of one against the other school is necessary, there should be caution against hasty application of one's frame of reference to an approach that does not intend to follow, or conform to, one's methodology. This should be done within the referential framework of the school critically evaluated, with due consideration for its methods and concepts. Similarly, as at present, there is no evidence to prove one or the other of these approaches as better, aside from personal choice. We can say so even if there is a strong paradigm shift towards the biological at present. A renaissance of scientific psychoanalysis coupled with a perceptive neurobiology which can translate those insights into testable hypotheses holds the greatest promise for psychiatry in the future. This suggests the need for unification of diverse appearing approaches to get a more comprehensive and enlightened worldview. It requires a highly integrative capacity. Just as a physicist thinks simultaneously in terms of particles and waves, a psychiatrist must think of motives, emotions and desires in the same breath as neurobiology, genetics and psychopharmacology. However, the integration must be attempted without destroying the internal cohesiveness of the individual schools. This will give a fair chance for polarization in which a single proper approach in psychiatry could emerge, which may be a conglomerate of diverse appearing approaches of today, or one which supersedes the rest. A synthesis of cognitive psychology and neuroscience offers the greatest promise at present.


Nightmares and disorders of dreaming.
Dreams occur during all stages of sleep. Nightmares are common. They can be associated with poor sleep and diminished daytime performance. Frequent nightmares are not related to underlying psychopathology in most children and in some "creative" adults. However, recurrent nightmares are the most defining symptom of post-traumatic stress disorder and may be associated with other psychiatric illnesses. Night terrors are arousal disorders that occur most often in children and usually occur early in the sleep period. Patients with rapid-eye-movement behavior disorder often present with nocturnal injury resulting from the acting out of dreams. Dream disorders may respond to medication, but behavioral treatment approaches have shown excellent results, particularly in patients with post-traumatic stress disorder and recurrent nightmares.


Screening for psychosocial distress among patients with cancer: implications for clinical practice, healthcare policy, and dissemination to enhance cancer survivorship.
Accreditation standards are at the forefront of evolving healthcare systems, setting metrics for high-quality care. Healthcare outcomes (health, experience, cost, provider satisfaction/burn out) are becoming mutual goals of the patient, provider, payer, and healthcare system. Achieving high-quality outcomes in cancer care necessitates collaboration among interdisciplinary teams of clinical providers, administrators, patient advocates, caregivers, and researchers. Dissemination and implementation science provides necessary frameworks to organize the efforts of these implementation teams, inclusive of identifying facilitators and barriers to implementation of accreditation standards. Since 2015, cancer distress screening has been mandated for continued cancer center accreditation by the American College of Surgeon's Commission on Cancer. Cancer centers have thus become real world implementation laboratories. We present the current context of distress screening, highlighting prior research and key areas of future research. We consider multiple levels of cancer care delivery and the use of interdisciplinary teams to help cancer center teams adopt, implement, and maintain efficient distress screening programs. Finally, we present a case study to identify methods for successful implementation of distress screening at one cancer center and then describe efficiencies that can be introduced using elements from human factors engineering, e- and m-health screening platforms, and community partnerships.


McKeever hemiarthroplasty of the knee in patients less than sixty years old.
Knee arthritis in the young patient is a challenging problem that may necessitate surgical treatment. We continue to perform hemiarthroplasty with a metallic tibial implant in selected young patients who, for various reasons, are not candidates for osteotomy, unicompartmental arthroplasty, or total knee arthroplasty. The purpose of the present study was to determine the minimum twelve-year results of this procedure in young patients. The original study group consisted of a consecutive series of twenty-four patients (twenty-six knees) who were managed with McKeever tibial hemiarthroplasty for the treatment of unicompartmental osteoarthritis of the knee. All patients were younger than sixty years of age at the time of the index procedure (average age, 44.6 years). During the study period, two patients died and one was lost to follow-up, leaving twenty-one patients (twenty-three knees) available for review. All patients were followed clinically for a minimum of twelve years or until revision. Knee Society knee and functional scores and Tegner scores were determined, and seven of the ten implants were evaluated radiographically. Thirteen knees were revised at an average of eight years after the index procedures. All thirteen knees had an uncomplicated revision to either a unicompartmental arthroplasty or total knee arthroplasty. Ten retained implants were available for clinical review after an average duration of follow-up of 16.8 years. The mean Knee Society knee scores, functional scores, and Tegner scores, available for nine of these ten knees, were 80, 97, and 4.2, respectively. We believe that the McKeever tibial hemiarthroplasty continues to be a reasonable surgical option for patients who are not candidates for osteotomy and are too young or too active for a unicompartmental or total knee arthroplasty.


Perception and adoption of a new agricultural technology: Evidence from a developing country.
Adoption of new agricultural technologies is always at the center of policy interest in developing countries. In reality, despite the visible benefits of many of the new agricultural technologies, including machinery and management practices, farmers either do not adopt them or it takes a long time to begin the adoption process and scaling up. To enhance the provision of irrigation using surface water and to enhance irrigation efficiency, Bangladesh has been trying to introduce the axial-flow-pump (AFP) appropriate for surface water irrigation, which can lift up to 55% more water, conditional on the water head, than a conventional centrifugal pump. Despite the visible benefits of the AFP, the uptake of the AFP for irrigation is low in the targeted zone of Bangladesh. The present study demonstrates that the new technology must be modified to adapt to local demand and specifications. Most importantly, the price of the new technology must be competitive with the prices of the existing available substitute technologies to ensure a rapid uptake and scaling up of this new agricultural technology.


Estimating the error rates of diagnostic tests.
It is often required to evaluate the accuracy of a new diagnostic test against a standard test with unknown error rates. If the two tests are applied simultaneously to the same individuals from two populations with different disease prevalences, then assuming conditional independence of the errors of the two tests, the error rates of both tests and the true prevalences in both populations can be estimated by a maximum likelihood procedure. Generalizations to several tests applied in several populations are also possible.


Sofia intermediate catheter and the SNAKE technique: safety and efficacy of the Sofia catheter without guidewire or microcatheter construct.
Neurointerventional surgeries (NIS) benefit from supportive endovascular constructs. Sofia is a soft-tipped, flexible, braided single lumen intermediate catheter designed for NIS. Sofia advancement from the cervical to the intracranial circulation without a luminal guidewire or microcatheter construct has not been described. To evaluate the efficacy and safety of the new Sofia Non-wire Advancement techniKE (SNAKE) for advancement of the Sofia into the cerebral circulation. Consecutive patients who underwent NIS using Sofia were identified. Patient information, SNAKE use, and patient outcome were determined from electronic medical records. Sofia advancement to the cavernous internal carotid artery or the V2/V3 segment junction of the vertebral artery was the primary outcome measure. Secondary outcomes included arterial vasospasm and arterial dissection. 263 Patients (181 females, 69%) who underwent a total of 305 NIS using Sofia were identified. SNAKE (SNAKE+) was used in 187 procedures (61%). Two hundred and ninety-three procedures (96%) were technically successful, which included 184 SNAKE+ NIS and 109 SNAKE- NIS. Primary outcome was achieved in all SNAKE+ procedures, but not in five SNAKE- procedures (2%). No arterial dissections were identified among 305 interventions. In the intracranial circulation, a single SNAKE+ patient (0.5%) had non-flow limiting arterial vasospasm involving the petrous internal carotid. Three SNAKE+ patients (1.6%) and one SNAKE- patient (0.8%) demonstrated external carotid artery branch artery vasospasm during dural arteriovenous fistula or facial arteriovenous malformation treatment. SNAKE is a safe and effective technique for Sofia advancement. Sofia is a highly effective and safe intermediate catheter for a variety of NIS.


Incidence, cardiovascular complications and mortality of hypertension by sex and ethnicity.
To compare ethnic and sex difference in the incidence of newly diagnosed hypertension, and subsequent risk of cardiovascular disease outcomes among South Asian, Chinese and white patients. We identified patients with newly diagnosed hypertension aged ≥20 years. Patients were followed for 1-9 years for all-cause mortality and cardiovascular disease with myocardial infarction, heart failure and stroke. Cox proportional hazard models stratified by sex and adjusted for age, median income and co-morbid conditions, were constructed to determine the independent association between ethnicity and the development of the combined cardiovascular endpoint as well as death. There were 39 175 South Asian (49.4% men, 34.4% age ≥65), 49 892 Chinese (48.1% men, 36.7% age ≥65) and 841 277 white (47.9% men, 38.8% age ≥65) patients with newly diagnosed hypertension. Age and sex adjusted incidence of hypertension was highest in South Asian patients and lowest in Chinese patients. Compared with white patients, South Asian and Chinese patients had a lower mortality (adjusted HR (aHR) 0.91 and 0.66) and risk of cardiovascular disease outcomes (aHR 0.94 and 0.49). Compared to men, women had significantly lower mortality (aHR: 0.83 for Chinese, 0.78 for South Asian and 0.77 for white) and cardiovascular disease outcomes (0.72 for Chinese, 0.63 for South Asian and 0.65 for white). South Asian patients had higher rates of hypertension compared to the other ethnic groups. South Asian and Chinese patients had a lower risk of death and developing cardiovascular outcomes compared to whites. Women with hypertension have a better prognosis than men regardless of ethnicity.


The transmitter release-site CaV2.2 channel cluster is linked to an endocytosis coat protein complex.
Synaptic vesicles (SVs) are triggered to fuse with the surface membrane at the presynaptic transmitter release site (TRSs) core by Ca2+ influx through nearby attached CaV2.2 channels [see accompanying paper: Khanna et al. (2007)Eur. J. Neurosci., 26, 547-559] and are then recovered by endocytosis. In this study we test the hypothesis that the TRS core is linked to an endocytosis-related protein complex. This was tested by immunostaining analysis of the chick ciliary ganglion calyx presynaptic terminal and biochemical analysis of synaptosome lysate, using CaV2.2 as a marker for the TRS. We noted that CaV2.2 clusters abut heavy-chain (H)-clathrin patches at the transmitter release face. Quantitative coimmunostaining analysis (ICA/ICQ method) demonstrated a strong covariance of release-face CaV2.2 staining with that for the AP180 and intersectin endocytosis adaptor proteins, and a moderate covariance with H- or light-chain (L)-clathrin and dynamin coat proteins, consistent with a multimolecular complex. This was supported by coprecipitation of these proteins with CaV2.2 from brain synaptosome lysate. Interestingly, the channel neither colocalized nor coprecipitated with the endocytosis cargo-capturing adaptor AP2, even though this protein both colocalized and coprecipitated with H-clathrin. Fractional recovery analysis of the immunoprecipitated CaV2.2 complex by exposure to high NaCl (approximately 1 m) indicated that AP180 and S-intersectin adaptors are tightly bound to CaV2.2 while L-intersectin, H- and L-clathrin and dynamin form a less tightly linked subcomplex. Our results are consistent with two distinct clathrin endocytosis complexes: an AP2-containing, remote, non-TRS complex and a specialised, AP2-lacking, TRS-associated subcomplex linked via a molecular bridge. The most probable role of this subcomplex is to facilitate SV recovery after transmitter release.


Mitigating Moral Distress in Dementia Care: Implications for Leaders in the Residential Care Sector.
In 2012, the World Health Organization estimated that the number of people living with dementia worldwide was approximately 35.6 million; they projected a doubling of this number by 2030, and tripling by 2050. Although the majority of people living with a dementia live in the community, residential facility care by nursing providers is a common part of the dementia journey in most countries. Previously published research confirms that caring for people living with dementia in such facilities often creates moral distress for nursing care providers. In this paper, the authors share additional findings from a two-year, two-phase, mixed methods study of moral distress as experienced by nursing caregivers of residents with dementia in residential care settings in a Western Canadian province. The findings relate to strategies to reduce moral distress in this caregiving group, with a particular focus on the role of supportive and responsive leadership. Important implications for practice and for leadership in the residential care sector are presented.


G protein-coupled receptors in human fat taste perception.
In contrast to carbohydrates and proteins, which are detected by specialized taste receptors in the forms of their respective building blocks, sugars, and L-amino acids, the third macronutrient, lipids, has until now not been associated with gustatory receptors. Instead, the recognition of fat stimuli was believed to rely mostly on textural, olfactory, and postingestive cues. During the recent years, however, research done mainly in rodent models revealed an additional gustatory component for the detection of long-chain fatty acids (LCFAs), the main taste-activating component of lipids. Concomitantly, a number of candidate fat taste receptors were proposed to be involved in rodent's gustatory fatty acid perception. Compared with rodent models, much less is known about human fat taste. In order to investigate the ability of the human gustatory system to respond to fat components, we performed sensory experiments with fatty acids of different chain lengths and derivatives thereof. We found that our panelists discriminated a "fatty" and an irritant "scratchy" taste component, with the "fatty" percept restricted to LCFAs. Using functional calcium-imaging experiments with the human orthologs of mouse candidate fat receptors belonging to the G protein-coupled receptor family, we correlated human sensory data with receptor properties characterized in vitro. We demonstrated that the pharmacological activation profile of human GPR40 and GPR120, 2 LCFA-specific receptors associated with gustatory fat perception in rodents, is inconsistent with the "scratchy" sensation of human subjects and more consistent with the percept described as "fatty." Expression analysis of GPR40 and GPR120 in human gustatory tissues revealed that, while the GPR40 gene is not expressed, GPR120 is detected in gustatory and nongustatory epithelia. On a cellular level, we found GPR120 mRNA and protein in taste buds as well as in the surrounding epithelial cells. We conclude that GPR120 may indeed participate in human gustatory fatty acid perception.


Innate immune responses to Listeria in vivo.
Listeria monocytogenes (Lm) is a foodborne bacterial pathogen that causes listeriosis, a severe infection that manifests as bacteremia and meningo-encephalitis mostly in immunocompromised individuals, and maternal-fetal infection. A critical pathogenic determinant of Lm relies on its ability to actively cross the intestinal barrier, disseminate systemically and cross the blood-brain and placental barriers. Here we illustrate how Lm both evades innate immunity, favoring its dissemination in host tissues, and triggers innate immune defenses that participate to its control.


Polaromonas naphthalenivorans sp. nov., a naphthalene-degrading bacterium from naphthalene-contaminated sediment.
Strain CJ2T, capable of growth on naphthalene as a sole carbon and energy source, was isolated from coal-tar-contaminated freshwater sediment. The Gram reaction of strain CJ2T was negative. The cells were non-spore-forming, non-motile cocci (without flagella). The isolate was found to be an aerobic heterotroph capable of utilizing glucose and other simple sugars. Growth was observed between 4 and 25 degrees C (optimum, 20 degrees C) and between pH 6.0 and 9.0 (optimum, pH 7.0-7.5). The G+C content of the genomic DNA was 61.5 mol% and the major quinone was ubiquinone-8. The peptidoglycan of strain CJ2T was determined as belonging to type A1-gamma, meso-diaminopimelic acid. The major fatty acids of strain CJ2T were 16:1omega7c (67.0%), 16:0 (19.6%), 18:1omega7c (approximately 7.9%) and 10:0 3-OH (approximately 2.5%). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Mycolic acid and glycolipids could not be detected. Comparative 16S rDNA analysis indicated that strain CJ2T is related to the family Comamonadaceae and that the nearest phylogenetic relative was Polaromonas vacuolata 34-PT (97.1% similarity). On the basis of the physiological and molecular properties, the naphthalene-degrading isolate was designated Polaromonas naphthalenivorans sp. nov. The type strain is CJ2T (=ATCC BAA-779T=DSM 15660T).


The NADPH organizers NoxO1 and p47phox are both mediators of diabetes-induced vascular dysfunction in mice.
NADPH oxidases are important sources of reactive oxygen species (ROS). Several Nox homologues are present together in the vascular system but whether they exhibit crosstalk at the activity level is unknown. To address this, vessel function of knockout mice for the cytosolic Nox organizer proteins p47phox, NoxO1 and a p47phox-NoxO1-double knockout were studied under normal condition and during streptozotocin-induced diabetes. In the mouse aorta, mRNA expression for NoxO1 was predominant in smooth muscle and endothelial cells, whereas p47phox was markedly expressed in adventitial cells comprising leukocytes and tissue resident macrophages. Knockout of either NoxO1 or p47phox resulted in lower basal blood pressure. Deletion of any of the two subunits also prevented diabetes-induced vascular dysfunction. mRNA expression analysis by MACE (Massive Analysis of cDNA ends) identified substantial gene expression differences between the mouse lines and in response to diabetes. Deletion of p47phox induced inflammatory activation with increased markers of myeloid cells and cytokine and chemokine induction. In contrast, deletion of NoxO1 resulted in an attenuated interferon gamma signature and reduced expression of genes related to antigen presentation. This aspect was also reflected by a reduced number of circulating lymphocytes in NoxO1-/- mice. ROS production stimulated by NoxO1 and p47phox limit endothelium-dependent relaxation and maintain blood pressure in mice. However, NoxO1 and p47phox cannot substitute each other despite their similar effect on vascular function. Deletion of NoxO1 induced an anti-inflammatory phenotype, whereas p47phox deletion rather elicited a hyper-inflammatory response.


Mixed arteriovenous malformation and capillary telangiectasia: a rare subset of mixed vascular malformations. Case report.
In this report, the authors discuss the case of a patient with a mixed cerebrovascular malformation in which an arteriovenous malformation (AVM) was associated with a capillary telangiectasia. Recent reports have contained reviews of various subsets of mixed malformations. To the authors' knowledge, however, this is the first report of a mixed vascular malformation with both arterial and capillary components. The patient underwent complete resection of the AVM after presenting with a clinical hemorrhage. She required a second operation to resect the capillary telangiectasia after new symptoms developed several months following the first procedure. The authors conclude that a mixed AVM-capillary telangiectasia is a rare but distinct entity.


Use of electronic health record systems by office-based pediatricians.
The American Recovery and Reinvestment Act of 2009 accelerated the implementation of electronic health records (EHRs) in pediatric offices. We sought to determine the prevalence and functionalities of EHRs, as well as pediatricians' perceptions of EHRs. An 8-page self-administered questionnaire was sent randomly to 1621 nonretired US members of the American Academy of Pediatrics from July to December 2012. Responses were compared with a similar survey in 2009. The percent of pediatricians, who are using EHRs, increased significantly from 58% in the 2009 survey to 79% in 2012. Only 31% used an EHR considered to have basic functionality, and only 14% used a fully functional EHR. Providers with equal or greater than 20% public insurance patients (threshold for meaningful use eligibility) were more likely to have an EHR. Solo/2-physician practices were least likely to have adopted an EHR. Younger physicians were more likely to consider an EHR important to quality care and perceived the presence of an EHR as more important in recruiting. The number of office-based pediatricians who are using an EHR has steadily risen to almost 80%. EHR cost and reduction in productivity remain serious concerns. Despite the widespread adoption of EHRs by pediatricians, only few use a basic or fully functional EHR and even fewer have added pediatric functionality. There is a role for the EHR certification process to advance functionalities used by pediatricians and to increase efficiency, data exchange capability, and general EHR functionality.


The road to independence: lived experiences of youth with chronic conditions and their parents compared.
To gain insight into the development of young persons with chronic conditions towards independence by comparing their lived experiences to those of their parents. Semi-structured interviews were held with 16 young persons (7 males, 9 females; 15-22 years) and one of their parents (n=16), asking about the young persons' daily lives and their development towards adulthood. Themes were deductively derived from the Skills for Growing Up framework, i.e. agency, living and daily activities, social and intimate relationships, education, work, leisure activities, transportation, and healthcare. Parents also reflected on how they dealt with the child's chronic condition. A paired thematic analysis was conducted. Parents were often less convinced that their children could act independently than the young persons themselves. They were concerned about them and tended to interfere with their daily lives, often to the annoyance of their children. Also, parents often perceived more barriers in social participation, while young persons were more positive. The perceptions of young persons and their parents clashed on living independently, intimate relationships, leisure activities, and healthcare. Young people might benefit from professional support in these domains to help them strengthen their autonomy and to prevent child-parent conflict and negative outcomes.


Familiarity Speeds Up Visual Short-term Memory Consolidation: Electrophysiological Evidence from Contralateral Delay Activities.
To test how preexisting long-term memory influences visual STM, this study takes advantage of individual differences in participants' prior familiarity with Pokémon characters and uses an ERP component, the contralateral delay activity (CDA), to assess whether observers' prior stimulus familiarity affects STM consolidation and storage capacity. In two change detection experiments, consolidation speed, as indexed by CDA fractional area latency and/or early-window (500-800 msec) amplitude, was significantly associated with individual differences in Pokémon familiarity. In contrast, the number of remembered Pokémon stimuli, as indexed by Cowan's K and late-window (1500-2000 msec) CDA amplitude, was significantly associated with individual differences in Pokémon familiarity when STM consolidation was incomplete because of a short presentation of Pokémon stimuli (500 msec, Experiment 2), but not when STM consolidation was allowed to complete given sufficient encoding time (1000 msec, Experiment 1). Similar findings were obtained in between-group analyses when participants were separated into high-familiarity and low-familiarity groups based on their Pokémon familiarity ratings. Together, these results suggest that stimulus familiarity, as a proxy for the strength of preexisting long-term memory, primarily speeds up STM consolidation, which may subsequently lead to an increase in the number of remembered stimuli if consolidation is incomplete. These findings thus highlight the importance of research assessing how effects on representations (e.g., STM capacity) are in general related to (or even caused by) effects on processes (e.g., STM consolidation) in cognition.


Thrombolytic and pharmacokinetic properties of chimeric tissue-type and urokinase-type plasminogen activators.
Chimeric molecules comprising the A-chain of tissue-type plasminogen activator (t-PA) and the catalytic domain of urokinase-type plasminogen activator (u-PA) have intact enzymatic characteristics of u-PA, partial fibrin-binding properties of t-PA, and thrombolytic properties in animal models comparable with but not superior to those of single-chain u-PA (scu-PA). Deletion of the finger and growth factor domains (t-PA-delta FE/scu-PA-e) in such chimeras further reduces their affinity for fibrin. A detailed investigation of the thrombolytic potency and the pharmacokinetics of t-PA and u-PA chimeras was performed in quantitative animal models for thrombolysis. In hamsters with pulmonary embolism, in rabbits with jugular vein thrombosis, and in baboons with femoral vein thrombosis, the thrombolytic potency (percent lysis per milligram of compound administered per kilogram of body weight) of t-PA-delta FE/scu-PA-e was significantly higher than that of recombinant scu-PA (rscu-PA, Saruplase) as shown by a maximal rate of 720 +/- 170% versus 45 +/- 5% lysis per milligram of compound per kilogram of body weight (mean +/- SEM, p less than 0.01) in hamsters, 210 +/- 18% versus 49 +/- 3% lysis per milligram of compound per kilogram of body weight (mean +/- SEM, p less than 0.01) in rabbits, and 310 +/- 73% versus 90 +/- 0.3% lysis per milligram of compound per kilogram of body weight (p less than 0.01) in baboons. However, the specific thrombolytic activity (percent lysis per microgram per milliliter steady-state plasma antigen level) of t-PA-delta FE/scu-PA-e was not significantly different from that of rscu-PA in hamsters (210 +/- 57% versus 160 +/- 27% lysis per microgram per milliliter antigen level) and was lower than that of rscu-PA in rabbits (37 +/- 4% versus 130 +/- 5% lysis per microgram per milliliter antigen level; p less than 0.01). In dogs with a combined femoral vein blood clot and a platelet-rich femoral arterial eversion graft thrombosis, 0.25 mg/kg body wt bolus injections of t-PA-delta FE/scu-PA-e produced significantly more venous clot lysis (90 +/- 5%, n = 10) than 0.25 mg/kg rscu-PA (26 +/- 3%, n = 10) (p less than 0.001) and, at the arterial side, more frequent (10 of 10 dogs versus three of 10 dogs) and more persistent (six of 10 dogs versus none of 10 dogs) recanalization (p = 0.002). After bolus injection in hamsters, rabbits, or baboons, t-PA-delta FE/scu-PA-e had a fourfold to sixfold longer initial half-life than rscu-PA and a slower plasma clearance of sixfold in hamsters, 10-fold in rabbits, and more than 10-fold in baboons. These results indicate that t-PA-delta FE/scu-PA-e has a markedly enhanced thrombolytic potency toward venous and arterial thrombi caused by a delayed in vivo clearance with relatively maintained specific thrombolytic activity. These properties suggest that the chimera may be clinically useful for thrombolytic therapy by bolus administration in patients with thromboembolic disease.


Spacer-length dependence of programmed -1 or -2 ribosomal frameshifting on a U6A heptamer supports a role for messenger RNA (mRNA) tension in frameshifting.
Programmed -1 ribosomal frameshifting is employed in the expression of a number of viral and cellular genes. In this process, the ribosome slips backwards by a single nucleotide and continues translation of an overlapping reading frame, generating a fusion protein. Frameshifting signals comprise a heptanucleotide slippery sequence, where the ribosome changes frame, and a stimulatory RNA structure, a stem-loop or RNA pseudoknot. Antisense oligonucleotides annealed appropriately 3' of a slippery sequence have also shown activity in frameshifting, at least in vitro. Here we examined frameshifting at the U6A slippery sequence of the HIV gag/pol signal and found high levels of both -1 and -2 frameshifting with stem-loop, pseudoknot or antisense oligonucleotide stimulators. By examining -1 and -2 frameshifting outcomes on mRNAs with varying slippery sequence-stimulatory RNA spacing distances, we found that -2 frameshifting was optimal at a spacer length 1-2 nucleotides shorter than that optimal for -1 frameshifting with all stimulatory RNAs tested. We propose that the shorter spacer increases the tension on the mRNA such that when the tRNA detaches, it more readily enters the -2 frame on the U6A heptamer. We propose that mRNA tension is central to frameshifting, whether promoted by stem-loop, pseudoknot or antisense oligonucleotide stimulator.


COPD uncovered: an international survey on the impact of chronic obstructive pulmonary disease [COPD] on a working age population.
Approximately 210 million people are estimated to have chronic obstructive pulmonary disease [COPD] worldwide. The burden of disease is known to be high, though less is known about those of a younger age. The aim of this study was to investigate the wider personal, economic and societal burden of COPD on a cross country working-age cohort. A cross-country [Brazil, China, Germany, Turkey, US, UK] cross-sectional survey methodology was utilised to answer the research questions. 2426 participants aged 45-67 recruited via a number of recruitment methods specific to each country completed the full survey. Inclusion criteria were a recalled physician diagnosis of COPD, a smoking history of > 10 pack years and the use of COPD medications in the previous 3 months prior to questioning. The survey included items from the validated Work Productivity and Activity Impairment [WPAI] scale and the EuroQoL 5 Dimension [EQ-5D] scale. Disease severity was measured using the 5-point MRC [Medical Research Council] dyspnoea scale as a surrogate measure. 64% had either moderate [n = 1012] or severe [n = 521] COPD, although this varied by country. 75% of the cohort reported at least one comorbid condition. Quality of life declined with severity of illness [mild, mean EQ-5D score = 0.84; moderate 0.58; severe 0.41]. The annual cost of healthcare utilisation [excluding treatment costs and diagnostic tests] per individual was estimated to be $2,364 [£1,500]. For those remaining in active employment [n: 677]: lost time from work cost the individual an average of $880 [£556] per annum and lifetime losses of $7,365 [£4,661] amounting to $596,000 [£377,000] for the cohort. 447 [~40%] of the working population had retired prematurely because of COPD incurring individual estimated lifetime income losses of $316,000 [£200,000] or a combined total of $141 m [£89.6 m]. As the mean age of retirees was 58.3 and average time since retirement was 4 years, this suggests the average age of retirement is around 54. This would mean a high societal and economic impact in all study countries, particularly where typical state retirement ages are higher, for example in Brazil, Germany and the UK [65] and the US [65,66,67], compared to Turkey [58 for women, 60 for men] and China [60]. Although generalisation across a broader COPD population is limited due to the varied participant recruitment methods, these data nevertheless suggest that COPD has significant personal, economic and societal burden on working age people. Further efforts to improve COPD diagnosis and management are required.


Placental secretome characterization identifies candidates for pregnancy complications.
Alterations in maternal physiological adaptation during pregnancy lead to complications, including abnormal birthweight and gestational diabetes. Maternal adaptations are driven by placental hormones, although the full identity of these is lacking. This study unbiasedly characterized the secretory output of mouse placental endocrine cells and examined whether these data could identify placental hormones important for determining pregnancy outcome in humans. Secretome and cell peptidome analyses were performed on cultured primary trophoblast and fluorescence-activated sorted endocrine trophoblasts from mice and a placental secretome map was generated. Proteins secreted from the placenta were detectable in the circulation of mice and showed a higher relative abundance in pregnancy. Bioinformatic analyses showed that placental secretome proteins are involved in metabolic, immune and growth modulation, are largely expressed by human placenta and several are dysregulated in pregnancy complications. Moreover, proof-of-concept studies found that secreted placental proteins (sFLT1/MIF and ANGPT2/MIF ratios) were increased in women prior to diagnosis of gestational diabetes. Thus, placental secretome analysis could lead to the identification of new placental biomarkers of pregnancy complications.


Testicular mixed germ cell tumors: a morphological and immunohistochemical study using stem cell markers, OCT3/4, SOX2 and GDF3, with emphasis on morphologically difficult-to-classify areas.
Stem cell markers, OCT3/4, and more recently SOX2 and growth differentiation factor 3 (GDF3), have been reported to be expressed variably in germ cell tumors. We investigated the immunohistochemical expression of these markers in different testicular germ cell tumors, and their utility in the differential diagnosis of morphologically difficult-to-classify components of these tumors. A total of 50 mixed testicular germ cell tumors, 43 also containing difficult-to-classify areas, were studied. In these areas, multiple morphological parameters were noted, and high-grade nuclear details similar to typical embryonal carcinoma were considered 'embryonal carcinoma-like high-grade'. Immunohistochemical staining for OCT3/4, c-kit, CD30, SOX2, and GDF3 was performed and graded in each component as 0, negative; 1+, 1-25%; 2+, 26-50%; and 3+, >50% positive staining cells. The different components identified in these tumors were seminoma (8), embryonal carcinoma (50), yolk sac tumor (40), teratoma (40), choriocarcinoma (3) and intra-tubular germ cell neoplasia, unclassified (35). By immunohistochemistry, the staining patterns were OCT3/4 -3+, all seminomas, embryonal carcinomas and intra-tubular germ cell neoplasia; SOX2 -3+, all embryonal carcinomas and -2 to 3+, 11/14 (79%) primitive neuroectodermal components in immature teratomas; GDF3 -2 to 3+, all yolk sac tumors, seminomas and intra-tubular germ cell neoplasia and 1 to 2+, 40/50 embryonal carcinomas. A total of 34/43 (79%) of difficult-to-classify areas stained 3+ for OCT3/4, CD30, and SOX2, similar to embryonal carcinoma. Among these areas, only 'embryonal carcinoma-like high-grade' nuclear details were significantly associated with such an immunophenotype. Thus, SOX2 is expressed in embryonal carcinoma and primitive neuroectoderm of teratoma, and unlike OCT3/4, not in intra-tubular germ cell neoplasia and seminoma. Therefore, it may be useful in the distinction of seminoma from embryonal carcinoma, and potentially in diagnosing early carcinomatous differentiation in seminoma. GDF3 positivity, in the absence of OCT3/4 and CD30, combined with morphological features, is helpful in the diagnosis of yolk sac tumor. 'Embryonal carcinoma-like high-grade' nuclear details are the most important morphological criterion for the diagnosis of embryonal carcinoma in difficult-to-classify areas.


RNA-Seq of Tumor-Educated Platelets Enables Blood-Based Pan-Cancer, Multiclass, and Molecular Pathway Cancer Diagnostics.
Tumor-educated blood platelets (TEPs) are implicated as central players in the systemic and local responses to tumor growth, thereby altering their RNA profile. We determined the diagnostic potential of TEPs by mRNA sequencing of 283 platelet samples. We distinguished 228 patients with localized and metastasized tumors from 55 healthy individuals with 96% accuracy. Across six different tumor types, the location of the primary tumor was correctly identified with 71% accuracy. Also, MET or HER2-positive, and mutant KRAS, EGFR, or PIK3CA tumors were accurately distinguished using surrogate TEP mRNA profiles. Our results indicate that blood platelets provide a valuable platform for pan-cancer, multiclass cancer, and companion diagnostics, possibly enabling clinical advances in blood-based "liquid biopsies".


A review of artificial pancreas technologies with an emphasis on bi-hormonal therapy.
Since the discovery of insulin, great progress has been made to improve the accuracy and safety of automated insulin delivery systems to help patients with type 1 diabetes achieve their treatment goals without causing hypoglycaemia. In recent years, bioengineering technology has greatly advanced diabetes management, with the development of blood glucose meters, continuous glucose monitors, insulin pumps and control systems for automatic delivery of one or more hormones. New insulin analogues have improved subcutaneous absorption characteristics, but do not completely eliminate the risk of hypoglycaemia. Insulin effect is counteracted by glucagon in non-diabetic individuals, while glucagon secretion in those with type 1 diabetes is impaired. The use of glucagon in the artificial pancreas is therefore a logical and feasible option for preventing and treating hypoglycaemia. However, commercially available glucagon is not stable in aqueous solution for long periods, forming potentially cytotoxic fibrils that aggregate quickly. Therefore, a more stable formulation of glucagon is needed for long-term use and storage in a bi-hormonal pump. In addition, a model of glucagon action in type 1 diabetes is lacking, further limiting the inclusion of glucagon into systems employing model-assisted control. As a result, although several investigators have been working to help develop bi-hormonal systems for patients with type 1 diabetes, most continue to utilize single hormone systems employing only insulin. This article seeks to focus on the attributes of glucagon and its use in bi-hormonal systems.


Activating Transcription Factor 5 Promotes Neuroblastoma Metastasis by Inducing Anoikis Resistance.
MYCN-amplified neuroblastoma often presents as a highly aggressive metastatic disease with a poor prognosis. Activating transcription factor 5 (ATF5) is implicated in neural cell differentiation and cancer cell survival. Here, we show that ATF5 is highly expressed in patients with stage 4 high-risk neuroblastoma, with increased expression correlating with a poorer prognosis. We demonstrated that ATF5 promotes the metastasis of neuroblastoma cell lines in vivo. Functionally, ATF5 depletion significantly reduced xenograft tumor growth and metastasis of neuroblastoma cells to the bone marrow and liver. Mechanistically, ATF5 endows tumor cells with resistance to anoikis, thereby increasing their survival in systemic circulation and facilitating metastasis. We identified the proapoptotic BCL-2 modifying factor (BMF) as a critical player in ATF5-regulated neuroblastoma anoikis. ATF5 suppresses BMF under suspension conditions at the transcriptional level, promoting anoikis resistance, whereas BMF knockdown significantly prevents ATF5 depletion-induced anoikis. Therapeutically, we showed that a cell-penetrating dominant-negative ATF5 peptide, CP-d/n-ATF5, inhibits neuroblastoma metastasis to the bone marrow and liver by inducing anoikis sensitivity in circulating tumor cells. Our study identified ATF5 as a metastasis promoter and CP-d/n-ATF5 as a potential antimetastatic therapeutic agent for neuroblastoma. This study shows that resistance to anoikis in neuroblastoma is mediated by ATF5 and offers a rationale for targeting ATF5 to treat metastatic neuroblastoma.


In vivo induction of CTL responses by recombinant adenylate cyclase of Bordetella pertussis carrying multiple copies of a viral CD8(+) T-cell epitope.
Bordetella pertussis adenylate cyclase toxin (ACT) is one of the few known protein toxins penetrating directly into the cytosol of target cells across their cytoplasmic membrane without the need for endocytosis. This capacity of ACT was recently exploited for in vivo delivery of single viral CD8(+) T-epitopes into MHC class I-presenting cells and induction of protective antiviral cytotoxic T-cell (CTL) responses. Here, we have explored the potential of the cell-invasive adenylate cyclase domain of the toxin to deliver larger antigens by evaluating the epitope-specific CTL responses induced by constructs bearing one to four copies of the CD8(+) T-epitope from the nucleoprotein of the lymphocytic choriomeningitis virus. The increase in the number of copies of the epitope was accompanied by a moderate decrease of the specific cell invasiveness of the ACT protein and did not lead to further enhancement of the level of induced epitope-specific CTL cells in mice, as compared to ACT with a single copy of the epitope. These results demonstrate the capacity of ACT to deliver larger heterologous antigens comprising several epitopes for antigenic presentation in vivo.