Efficient energies and algorithms for parametric snakes.
Parametric active contour models are one of the preferred approaches for image segmentation because of their computational efficiency and simplicity. However, they have a few drawbacks which limit their performance. In this paper, we identify some of these problems and propose efficient solutions to get around them. The widely-used gradient magnitude-based energy is parameter dependent; its use will negatively affect the parametrization of the curve and, consequently, its stiffness. Hence, we introduce a new edge-based energy that is independent of the parameterization. It is also more robust since it takes into account the gradient direction as well. We express this energy term as a surface integral, thus unifying it naturally with the region-based schemes. The unified framework enables the user to tune the image energy to the application at hand. We show that parametric snakes can guarantee low curvature curves, but only if they are described in the curvilinear abscissa. Since normal curve evolution do not ensure constant arc-length, we propose a new internal energy term that will force this configuration. The curve evolution can sometimes give rise to closed loops in the contour, which will adversely interfere with the optimization algorithm. We propose a curve evolution scheme that prevents this condition.


Variants of ESR1, APOE, LPL and IL-6 loci in young healthy subjects: association with lipid status and obesity.
BMI was increased (>25) in 22% of young healthy subjects. Increased cholesterol values (>5.0 mmol/L) were found in 23% of subjects, LDL-C (>3.0 mmol/L) in 23%, triglycerides (>1.7 mmol/L) in 11% of subjects. We found statistically significant differences in subjects' weight (p = 0.015), BMI (p = 0.023), and waist-hip ratio (WHR) (p = 0.015) in regard to their diet type; subjects with Mediterranean diet had the lowest values compared to those on continental and mixed diet. Significant associations were found for: LPL genetic polymorphic variant and abdominal obesity (p = 0.013), APO epsilon4 allele and hypercholesterolemia (p = 0.003), and ESR1-TA long allele and hypercholesterolemia (p = 0.011). Human obesity is a multifactorial syndrome influenced also by genetic factors. Among gene variants found to be involved in body weight regulation and development of obesity, particular attention has been paid to polymorphisms in genes associated with obesity-related metabolic disorders. We explored the association of genetic polymorphisms of: estrogen receptor alpha (ESR1-TA repeats); interleukin-6 (IL-6 G-174C); apolipoprotein E (APO epsilon2, epsilon3, epsilon4); lipoprotein lipase Pvu II (LPL P+/-), with clinical variables: gender, age, body mass index (BMI), diet type and biological variables: triglycerides, cholesterol, HDL-C, LDL-C, CRP, homocysteine, urate, and glucose in 105 healthy young subjects (20-35 yrs) of Croatian origin. Genotyping of IL-6, LPL was performed by PCR-RFLP, of APOE by real-time PCR, and of ESR1 by PCR and capillary electrophoresis. Association analyses were performed of alleles and genotypes with biological variables. ESR-1, LPL, and APO E genetic polymorphic variants could represent predictive genetic risk markers for obesity-related metabolic disorders in young healthy subjects. Mediterranean type of diet is also an important protective factor against abdominal obesity.


Teacher support of adherence for adolescents with type 1 diabetes: preferred teacher support behaviors and youths' perceptions of support.
The purpose of this study is to examine the preferences of adolescents with type 1 diabetes regarding the role of teacher support and perceptions associated with the most supportive and least supportive teachers, as well as to examine the effects of child age and gender on these relationships. Method This cross-sectional questionnaire-based study included 102 adolescents with type 1 diabetes recruited through the outpatient treatment clinic at a Midwestern children's hospital. Adolescents completed demographic questionnaires and questionnaires assessing the role of teacher support. Outcome measures included the newly developed Ideal Teacher Questionnaire, which measured adolescents' preferences for teacher support, and the Diabetes Teacher Support Questionnaire, which evaluated adolescents' perceived frequency of teacher support behaviors and feelings about this support. Youth rated their most supportive teacher as engaging more frequently in support behaviors than their least supportive teacher. Students who expressed a preference for an actively involved teacher had more positive feelings about the support they received from teachers and perceived a higher level of support from their most supportive teacher. Girls perceived a greater frequency of support behaviors than boys and felt more positive about these behaviors. Younger adolescents reported a higher frequency of teacher support behaviors and felt that these behaviors were more supportive. There are consistent interpersonal differences in adolescents' desires for teacher support of diabetes self-care. It may be important to consider adolescents' preferences for diabetes-related teacher support.


Estradiol regulates brown adipose tissue thermogenesis via hypothalamic AMPK.
Estrogens play a major role in the modulation of energy balance through central and peripheral actions. Here, we demonstrate that central action of estradiol (E2) inhibits AMP-activated protein kinase (AMPK) through estrogen receptor alpha (ERα) selectively in the ventromedial nucleus of the hypothalamus (VMH), leading to activation of thermogenesis in brown adipose tissue (BAT) through the sympathetic nervous system (SNS) in a feeding-independent manner. Genetic activation of AMPK in the VMH prevented E2-induced increase in BAT-mediated thermogenesis and weight loss. Notably, fluctuations in E2 levels during estrous cycle also modulate this integrated physiological network. Together, these findings demonstrate that E2 regulation of the VMH AMPK-SNS-BAT axis is an important determinant of energy balance and suggest that dysregulation in this axis may account for the common changes in energy homeostasis and obesity linked to dysfunction of the female gonadal axis.


Differential filtering of two presynaptic depression mechanisms.
The filtering of input signals carried out at synapses is key to the information processing performed by networks of neurons. Two forms of presynaptic depression, vesicle depletion and G-protein inhibition of Ca2+ channels, can play important roles in the presynaptic processing of information. Using computational models, we demonstrate that these two forms of depression filter information in very different ways. G-protein inhibition acts as a high-pass filter, preferentially transmitting high-frequency input signals to the postsynaptic cell, while vesicle depletion acts as a low-pass filter. We examine how these forms of depression separately and together affect the steady-state postsynaptic responses to trains of stimuli over a range of frequencies. Finally, we demonstrate how differential filtering permits the multiplexing of information within a single impulse train.


Characteristics of Cutaneous Lymphomas in Korea According to the New WHO-EORTC Classification: Report of a Nationwide Study.
Previously, cutaneous lymphomas were classified according to either the European Organization for the Research and Treatment of Cancer (EORTC) or the World Health Organization (WHO) classification paradigms. The aim of this study was to determine the relative frequency of Korean cutaneous lymphoma according to the new WHO-EORTC classification system. A total of 517 patients were recruited during a recent 5 year-period (2006-2010) from 21 institutes and classified according to the WHO-EORTC criteria. The patients included 298 males and 219 females, and the mean age at diagnosis was 49 years. The lesions preferentially affected the trunk area (40.2%). The most frequent subtypes in order of decreasing prevalence were mycosis fungoides (22.2%), peripheral T-cell lymphoma (17.2%), CD30+ T-cell lymphoproliferative disorder (13.7%), and extranodal natural killer/T (NK/T) cell lymphoma, nasal type (12.0%). Diffuse large B-cell lymphoma accounted for 11.2% of cases, half of which were secondary cutaneous involvement; other types of B-cell lymphoma accounted for less than 1% of cases. In comparison with data from Western countries, this study revealed relatively lower rates of mycosis fungoides and B-cell lymphoma in Korean patients, as well as higher rates of subcutaneous panniculitis-like T-cell lymphoma and NK/T cell lymphoma.


Overt and 'subclinical' hypothyroidism in women.
Thyroid disease in general, and hypothyroidism in particular, are very common in women. In the USA, the most common cause of primary thyroid deficiency is on an autoimmune basis due to lymphocytic (Hashimoto) thyroiditis. Because there are thyroid hormone receptors in virtually every tissue of the body, the manifestations of hypothyroidism are varied, but problems with abnormal menses, conception, fertility, and pregnancy can be especially troubling in young women. The single most important diagnostic test is measurement of serum thyrotropin (TSH). The overwhelming majority of patients with hypothyroidism are treated with a single daily dose of synthetic levothyroxine with the goal of therapy being restoration of a normal metabolic state with return of the TSH level down to the range of 0.5 to 1.5 mlU/L. "Subclinical" hypothyroidism refers to those patients with early or mild thyroid hypofunction manifested as slight elevations of thyrotropin (approximately 4-10 mlU/L) although serum thyroxine (T4) and triiodothyronine (T3) levels are within their reference ranges. The entity is somewhat controversial in regard to its consequences if left untreated, and whether or not we should be screening patients, at least susceptible populations, for the condition. Reports indicate an association between subclinical hypothyroidism and poor outcomes of pregnancy, as well as dyslipidemias, atherogenesis, and increased mortality in the long term. We believe these consequences are sufficiently compelling to warrant screening and treatment with levothyroxine when found to halt progression to overt hypothyroidism, and improve symptoms, pregnancy outcomes, lipid abnormalities, and cardiovascular function. Obstetricians & Gynecologists, Family Physicians. After completion of this article, the reader should be able to recall that hypothyroidism is a common disease in women, has many protean manifestations, and can be successfully diagnosed and treated; explain that the condition of subclinical hypothyroidism can be diagnosed and if treated can prevent many untoward complications; and state that there should be heightened awareness of the disease so that proper screening tests can be performed.


Genetic admixture in multidimensional environmental space: asymmetrical niche similarity promotes gene flow in armadillos (Dasypus novemcinctus).
We unite genetic data with a robust test of niche divergence to test the hypothesis that patterns of gene flow between two lineages of the nine-banded armadillo are influenced by their climatic niches. We collected Geographical Information System (GIS) data on climate using locality information from 111 individuals from two lineages that had associated genetic material. We tested whether niches of these lineages were more conserved or divergent than the background environments of their geographic ranges and found evidence for niche conservatism on two axes and no evidence for divergence on any axis. To address the role of niche similarity in gene flow, we genotyped the 111 individuals at five microsatellite loci and tested whether admixed individuals tended to be located in parts of multidimensional environmental space (E-space) shared between the two lineages. We observed an asymmetrical pattern of overlap, in which the West lineage's E-space was almost completely included inside East lineage's E-space. Genetic admixture levels were significantly higher in the West lineage and, for both lineages, in shared portions of E-space. This suggests that niche similarity can facilitate gene flow among disjunct groups with moderate-to-good dispersal capabilities, contrasting with the prevailing view of niche conservatism as a diversifying force.


Approaches to the induction of HIV broadly neutralizing antibodies.
A vaccine that elicits antibody responses that can neutralize the diversity of HIV clades has not yet been achieved, and is a major focus of HIV vaccine research. Here, we provide an update on the barriers to eliciting such antibodies, and how advances in immunogen design may circumvent these roadblocks, focusing on data published in the last year. Studies of how broadly neutralizing antibodies (bNAbs) develop in HIV-infected donors continue to produce key insights, suggesting that for some viral targets there are common pathways to developing breadth. Germline-targeting strategies, that aim to recruit rare precursors of bNAbs, have shown promise in immunogenicity studies, and structural biology has led to advances in immunogen design. Mapping of strain-specific tier 2 vaccine responses has highlighted the challenges that remain in driving antibodies toward breadth. Elucidation of the HIV envelope structure, together with an understanding of how bNAbs emerge in vivo has guided the design of new immunogens and vaccine strategies that show promise for eliciting protective antibodies.


Potentiation of amyotrophic lateral sclerosis (ALS)-associated TDP-43 aggregation by the proteasome-targeting factor, ubiquilin 1.
TDP-43 (43-kDa TAR DNA-binding domain protein) is a major constituent of ubiquitin-positive cytoplasmic aggregates present in neurons of patients with fronto-temporal lobular dementia and amyotrophic lateral sclerosis (ALS). The pathologic significance of TDP-43 aggregation is not known; however, dominant mutations in TDP-43 cause a subset of ALS cases, suggesting that misfolding and/or altered trafficking of TDP-43 is relevant to the disease process. Here, we show that the presenilin-binding protein ubiquilin 1 (UBQLN) plays a role in TDP-43 aggregation. TDP-43 interacted with UBQLN both in yeast and in vitro, and the carboxyl-terminal ubiquitin-associated domain of UBQLN was both necessary and sufficient for binding to polyubiquitylated forms of TDP-43. Overexpression of UBQLN recruited TDP-43 to detergent-resistant cytoplasmic aggregates that colocalized with the autophagosomal marker, LC3. UBQLN-dependent aggregation required the UBQLN UBA domain, was mediated by non-overlapping regions of TDP-43, and was abrogated by a mutation in UBQLN previously linked to Alzheimer disease. Four ALS-associated alleles of TDP-43 also coaggregated with UBQLN, and the extent of aggregation correlated with in vitro UBQLN binding affinity. Our findings suggest that UBQLN is a polyubiquitin-TDP-43 cochaperone that mediates the autophagosomal delivery and/or proteasome targeting of TDP-43 aggregates.


A method for super-resolved CARS microscopy with structured illumination in two dimensions.
We propose a structured illumination scheme for achieving widefield coherent anti-Stokes Raman scattering (CARS) microscopy with a resolution surpassing the diffraction limit in two dimensions (2D). By acquiring a set of coherent images of a sample with third-order nonlinear susceptibility illuminated by the phase-matched excitation field of square lattice patterns, a 2D super-resolution CARS image can be reconstructed. We derive a theoretical framework to describe the coherent image formation and reconstruction scheme for this structured illumination CARS imaging system and carry out numerical simulations to investigate its imaging performance. The results demonstrate that our method promises a particular benefit on CARS microscopy by adding the super-resolution capability to improve its 2D spatial resolution by a factor of approximately three.


Mind-body treatments for the pain-fatigue-sleep disturbance symptom cluster in persons with cancer.
Co-occurring pain, fatigue, and sleep disturbance comprise a common symptom cluster in patients with cancer. Treatment approaches that target the cluster of symptoms rather than just a single symptom need to be identified and tested. To synthesize evidence regarding mind-body interventions that have shown efficacy in treating two or more symptoms in the pain-fatigue-sleep disturbance cancer symptom cluster. A literature search was conducted using CINAHL, Medline, and PsychInfo databases through March 2009. Studies were categorized based on the type of mind-body intervention (relaxation, imagery/hypnosis, cognitive-behavioral therapy/coping skills training [CBT/CST], meditation, music, and virtual reality), and a preliminary review was conducted with respect to efficacy for pain, fatigue, and sleep disturbance. Mind-body interventions were selected for review if there was evidence of efficacy for at least two of the three symptoms. Forty-three studies addressing five types of mind-body interventions met criteria and are summarized in this review. Imagery/hypnosis and CBT/CST interventions have produced improvement in all the three cancer-related symptoms individually: pain, fatigue, and sleep disturbance. Relaxation has resulted in improvements in pain and sleep disturbance. Meditation interventions have demonstrated beneficial effects on fatigue and sleep disturbance. Music interventions have demonstrated efficacy for pain and fatigue. No trials were found that tested the mind-body interventions specifically for the pain-fatigue-sleep disturbance symptom cluster. Efficacy studies are needed to test the impact of relaxation, imagery/hypnosis, CBT/CST, meditation, and music interventions in persons with cancer experiencing concurrent pain, fatigue, and sleep disturbance. These mind-body interventions could help patients manage all the symptoms in the cluster with a single treatment strategy.


Motion and vision. I. Stabilized images of stationary gratings.
To demonstrate that eye movements have profound effects on the sine-wave contrast threshold, the author uses a new method of stabilizing the retinal image, in which the Purkinje reflections from the eye move the stimulus pattern displayed on a CRT screen. Calibration of this compensatory motion is very critical; a gain error greater than 1% may produce significant destablization. Under optimum conditions, image stabilization elevates the subject's contrast threshold by a factor of about 20; it also produces after-images with resolution greater than 12 c/deg. These results compare favorably with those obtained by other methods.


Do infants fed from bottles lack self-regulation of milk intake compared with directly breastfed infants?
How breastfeeding reduces the risk of childhood obesity is unclear, and 1 hypothesis pertains to the ability of breastfed infants to self-regulate. We studied whether infants' self-regulation of milk intake is affected by feeding mode (bottle versus breast) and the type of milk in the bottle (formula versus expressed breast milk). Participants in the 2005-2007 Infant Feeding Practices Study II received monthly questionnaires during their infant's first year, and compete data were available for 1250 infants. We tested the impact of feeding mode and type of milk during early infancy on self-regulation during late infancy. Although only 27% of infants fed exclusively at the breast in early infancy emptied the bottle or cup in late infancy, 54% of infants who were fed both at the breast and by bottle did so, and 68% of those who were fed only by bottle did so. Multivariate regression analysis indicated that infants who were bottle-fed more intensively early in life were approximately 71% or 2 times more likely to empty the bottle or cup later in life than those who were bottle-fed less intensively ((1/3)-(2/3) or (2/3) of milk feeds given by bottle versus < (1/3) of milk feeds). When feeding formula and expressed milk were considered separately, similar dose-response relationships were observed. Infants who are bottle-fed in early infancy are more likely to empty the bottle or cup in late infancy than those who are fed directly at the breast. Bottle-feeding, regardless of the type of milk, is distinct from feeding at the breast in its effect on infants' self-regulation of milk intake.


A synthetic dodecanolide library for the identification of putative semiochemicals emitted by mantellid frogs.
Aliphatic macrolides are used by various organisms as semiochemicals or pheromones, e.g. by the endemic Madagascan frog family Mantellinae. Volatile, methyl-branched aliphatic macrolides occur in various species, but their identification by MS is difficult, while isolation is not possible due to the low amounts produced. Therefore, we synthesized a library of methyl-branched 11-dodecanolides to identify unknown macrolides secreted by Gephyromantis moseri, a mantelline frog restricted to the rainforest and characterized by largely terrestrial habitats. The syntheses used metal-organic epoxide opening and ring-closing-metathesis as key steps. All library members 2-, 4-, 6-, 8-, and 10-methyl-11-dodecanolide were thus identified as part of the secretion by comparison of their mass spectra and their gas chromatographic retention indices. These compounds comprise new natural products.


Effect of benazepril on heart rate turbulence in patients with dilated cardiomyopathy.
1. Heart rate (HR) turbulence describes short-term sinus rhythmic fluctuation after a single premature ventricular beat. Turbulence onset (TO) and turbulence slope (TS) are two essential parameters in HR turbulence. Turbulence onset and TS have been used to evaluate cardiac autonomic nerve function. 2. In the present study, we measured the HR turbulence in dilated cardiomyopathy (DCM) and determined the possible role of benazepril, an angiotensin-converting enzyme inhibitor (ACEI), on these parameters. There were three groups: control, DCM and DCM treated with benazepril. The control group consisted of normal subjects with PVB, but no structural heart disease. Ambulatory electrocardiogram, blood pressure and echocardiography were analysed. 3. There was an increase in TO and a decrease in TS in DCM patients. Benazepril treatment (10 mg/day, p.o.) reduced those changes. There were no significant differences in blood pressure and left ventricular ejection fraction (LVEF) between DCM patients and DCM patients treated with benazepril. 4. Linear regression analysis showed that TO was negatively correlated with LVEF, whereas TS was positively correlated with LVEF, in the DCM group. After benazepril treatment, the correlations between TO and TS and LVEF disappeared. 5. It is concluded that the TO and TS of HR turbulence are altered in patients with DCM. These alterations indicate a dysfunction of the autonomic control of cardiac electrophysiology in DCM patients. Although TO and TS are correlated with LVEF in DCM patients, the effect of benazepril in improving HR turbulence parameters is not a result of its action on heart function, which suggests a new beneficial effect of ACEI in the treatment of DCM patients.


A P element containing suppressor of hairy-wing binding regions has novel properties for mutagenesis in Drosophila melanogaster.
P elements are widely used as insertional mutagens to tag genes, facilitating molecular cloning and analyses. We modified a P element so that it carried two copies of the suppressor of Hairy-wing [su(Hw)] binding regions isolated from the gypsy transposable element. This transposon was mobilized, and the genetic consequences of its insertion were analyzed. Gene expression can be altered by the su(Hw) protein as a result of blocking the interaction between enhancer/silencer elements and their promoter. These effects can occur over long distances and are general. Therefore, a composite transposon (SUPor-P for suppressor-P element) combines the mutagenic efficacy of the gypsy element with the controllable transposition of P elements. We show that, compared to standard P elements, this composite transposon causes an expanded repertoire of mutations and produces alleles that are suppressed by su(Hw) mutations. The large number of heterochromatic insertions obtained is unusual compared to other insertional mutagenesis procedures, indicating that the SUPor-P transposon may be useful for studying the structural and functional properties of heterochromatin.


A fully integrated high-Q Whispering-Gallery Wedge Resonator.
Microresonator devices which possess ultra-high quality factors are essential for fundamental investigations and applications. Microsphere and microtoroid resonators support remarkably high Q's at optical frequencies, while planarity constrains preclude their integration into functional lightwave circuits. Conventional semiconductor processing can also be used to realize ultra-high-Q's with planar wedge-resonators. Still, their full integration with side-coupled dielectric waveguides remains an issue. Here we show the full monolithic integration of a wedge-resonator/waveguide vertically-coupled system on a silicon chip. In this approach the cavity and the waveguide lay in different planes. This permits to realize the shallow-angle wedge while the waveguide remains intact, allowing therefore to engineer a coupling of arbitrary strength between these two. The precise size-control and the robustness against post-processing operation due to its monolithic integration makes this system a prominent platform for industrial-scale integration of ultra-high-Q devices into planar lightwave chips.


Drivers of salamander extirpation mediated by Batrachochytrium salamandrivorans.
The recent arrival of Batrachochytrium salamandrivorans in Europe was followed by rapid expansion of its geographical distribution and host range, confirming the unprecedented threat that this chytrid fungus poses to western Palaearctic amphibians. Mitigating this hazard requires a thorough understanding of the pathogen's disease ecology that is driving the extinction process. Here, we monitored infection, disease and host population dynamics in a Belgian fire salamander (Salamandra salamandra) population for two years immediately after the first signs of infection. We show that arrival of this chytrid is associated with rapid population collapse without any sign of recovery, largely due to lack of increased resistance in the surviving salamanders and a demographic shift that prevents compensation for mortality. The pathogen adopts a dual transmission strategy, with environmentally resistant non-motile spores in addition to the motile spores identified in its sister species B. dendrobatidis. The fungus retains its virulence not only in water and soil, but also in anurans and less susceptible urodelan species that function as infection reservoirs. The combined characteristics of the disease ecology suggest that further expansion of this fungus will behave as a 'perfect storm' that is able to rapidly extirpate highly susceptible salamander populations across Europe.


Meningeal melanomatosis: a challenge for timely diagnosis.
Neoplastic meningitis is a central nervous system complication that occurs in 3-5% of patients with cancer. Although most commonly seen in patients with disseminated disease, in a small percentage of patients, it may be the initial manifestation of cancer or even primitive in origin. In the absence of cancer history, the diagnosis of neoplastic meningitis may be challenging even for expert neurologists. Prognosis is poor, with a median overall survival of weeks from diagnosis. In the retrospective study herein, we described three cases of meningeal melanomatosis in patients without previous cancer history. The patients were diagnosed with significant delay (17 to 47 weeks from symptom onset) mainly due to the deferral in performing the appropriate testing. Even when the diagnosis was suspected, investigations by MRI, cerebrospinal fluid, or both proved at times unhelpful for confirmation. Prognosis was dismal, with a median survival of 4 weeks after diagnosis. Our observations are a cue to analyze the main pitfalls in the diagnosis of neoplastic meningitis in patients without cancer history and emphasize key elements that may facilitate early diagnosis.


Soil microbial community structure across a thermal gradient following a geothermal heating event.
In this study microbial species diversity was assessed across a landscape in Yellowstone National Park, where an abrupt increase in soil temperature had occurred due to recent geothermal activity. Soil temperatures were measured, and samples were taken across a temperature gradient (35 to 65 degrees C at a 15-cm depth) that spanned geothermally disturbed and unimpacted soils; thermally perturbed soils were visually apparent by the occurrence of dead or dying lodgepole pine trees. Changes in soil microbial diversity across the temperature gradient were qualitatively assessed based on 16S rRNA sequence variation as detected by denaturing gradient gel electrophoresis (DGGE) using both ribosomal DNA (rDNA) and rRNA as PCR templates and primers specific for the Bacteria or Archaea domain. The impact of the major heating disturbance was apparent in that DGGE profiles from heated soils appeared less complex than those from the unaffected soils. Phylogenetic analysis of a bacterial 16S rDNA PCR clone library from a recently heated soil showed that a majority of the clones belonged to the Acidobacterium (51%) and Planctomyces (18%) divisions. Agar plate counts of soil suspensions cultured on dilute yeast extract and R2A agar media incubated at 25 or 50 degrees C revealed that thermophile populations were two to three orders of magnitude greater in the recently heated soil. A soil microcosm laboratory experiment simulated the geothermal heating event. As determined by both RNA- and DNA-based PCR coupled with DGGE, changes in community structure (marked change in the DGGE profile) of soils incubated at 50 degrees C occurred within 1 week and appeared to stabilize after 3 weeks. The results of our molecular and culture data suggest that thermophiles or thermotolerant species are randomly distributed in this area within Yellowstone National Park and that localized thermal activity selects for them.


Violence and touch deprivation in adolescents.
The increasing incidence of violence among children and adolescents highlights the importance of identifying at-risk profiles as well as assessing interventions for preventing violence. Empirical research has suggested behavioral, central nervous system, and neurotransmitter/neurohormone dysregulation in violent individuals, including (1) an underaroused central nervous system characterized by right frontal electroencephalogram (EEG) hypoactivation, and (2) a neurotransmitter/neurohormone profile of lower norepinephrine, serotonin, and cortisol, and elevated dopamine and testosterone. The literature also suggests a disproportionate incidence of physical abuse and neglect or the lack of positive physical contact in violent individuals. In the studies we have conducted to date, there has been a relatively high incidence of anger and aggression in high school samples, even those that were relatively advantaged, as well as high levels of depression (one standard deviation above the mean), suggesting significant disturbance in these youth. Adolescents with these profiles also had less optimal relationships with their families, used illicit drugs more frequently, had inferior academic performance, and had higher depression scores. In our cross-cultural comparisons, preschoolers and adolescents were less physically affectionate and more aggressive in the United States versus France. Further, the U.S. youth received less physical affection as preschoolers, and as adolescents they engaged in more self-stimulating behaviors, perhaps to compensate for receiving less physical affection from their parents and peers. This supports the notion that less physical affection (or more physical neglect) can contribute to greater aggression. Massage therapy has been effective with violent adolescents, perhaps because the physical stimulation reduced their dopamine levels and increased their serotonin levels. Their aggressive behavior decreased and their empathetic behavior increased. These preliminary data need to be replicated in a larger sample with a more comprehensive set of measures in the context of identifying a diagnostic profile.


Spatiotemporal analysis of sensor logs using growth ring maps.
Spatiotemporal analysis of sensor logs is a challenging research field due to three facts: a) traditional two-dimensional maps do not support multiple events to occur at the same spatial location, b) three-dimensional solutions introduce ambiguity and are hard to navigate, and c) map distortions to solve the overlap problem are unfamiliar to most users. This paper introduces a novel approach to represent spatial data changing over time by plotting a number of non-overlapping pixels, close to the sensor positions in a map. Thereby, we encode the amount of time that a subject spent at a particular sensor to the number of plotted pixels. Color is used in a twofold manner; while distinct colors distinguish between sensor nodes in different regions, the colors' intensity is used as an indicator to the temporal property of the subjects' activity. The resulting visualization technique, called Growth Ring Maps, enables users to find similarities and extract patterns of interest in spatiotemporal data by using humans' perceptual abilities. We demonstrate the newly introduced technique on a dataset that shows the behavior of healthy and Alzheimer transgenic, male and female mice. We motivate the new technique by showing that the temporal analysis based on hierarchical clustering and the spatial analysis based on transition matrices only reveal limited results. Results and findings are cross-validated using multidimensional scaling. While the focus of this paper is to apply our visualization for monitoring animal behavior, the technique is also applicable for analyzing data, such as packet tracing, geographic monitoring of sales development, or mobile phone capacity planning.


Discovery of multi-drug resistant, MCR-1 and ESBL-coproducing ST117 Escherichia coli from diseased chickens in northeast China.
An endemic multi-drug resistant ST117 E. coli isolate coproducing MCR-1 and 3 ESBL loci was, for the first time, detected from diseased chicken, Liaoning Province, in Northeast China, from 2011 to 2012. Whole-genome sequencing revealed 5 unique plasmids, namely pHXH-1, pHXH-2, pHXH-3, pHXH-4 and pHXH-5). Among them, pHXH1 and pHXH4 encode ESBL, and pHXH-5 mediates MCR-1 colistin resistance. The results indicate that the potentially-national dissemination of MCR-1-positive pathogens with pan-drug resistance proceeds via food chains.


How experts gain influence.
In theory, the risk management groups of two British banks--Saxon and Anglo--had the same influence in their organizations. But in practice, they did not: Saxon's was engaged in critical work throughout the bank, while Anglo's had little visibility outside its areas of expertise. In their study of these two financial institutions, the authors identified four competencies--trailblazing, toolmaking, teamwork, and translation--that help functional leaders or groups compete for top management's limited attention and increase their impact. Anglo's risk managers were strong in only some of the competencies, but Saxon's were strong in all four. They consistently scanned the internal and external environment for important issues to which they could apply a risk management perspective (trailblazing) and then developed tools--such as quarterly risk reports--that spread their expertise (toolmaking). While controlling the tools' design and implementation, the risk managers incorporated business managers' insights (teamwork) and made sure everyone could understand the findings (translation). Ultimately, experts' roles must fit the organization's strategy and structural needs. In some situations, functional experts can raise their profile by cultivating just two of the competencies. But those who are strong in all four are likely to be the most influential.


Classification of Melanocytic Lesions in Selected and Whole-Slide Images via Convolutional Neural Networks.
Whole-slide images (WSIs) are a rich new source of biomedical imaging data. The use of automated systems to classify and segment WSIs has recently come to forefront of the pathology research community. While digital slides have obvious educational and clinical uses, their most exciting potential lies in the application of quantitative computational tools to automate search tasks, assist in classic diagnostic classification tasks, and improve prognosis and theranostics. An essential step in enabling these advancements is to apply advances in machine learning and artificial intelligence from other fields to previously inaccessible pathology datasets, thereby enabling the application of new technologies to solve persistent diagnostic challenges in pathology. Here, we applied convolutional neural networks to differentiate between two forms of melanocytic lesions (Spitz and conventional). Classification accuracy at the patch level was 99.0%-2% when applied to WSI. Importantly, when the model was trained without careful image curation by a pathologist, the training took significantly longer and had lower overall performance. These results highlight the utility of augmented human intelligence in digital pathology applications, and the critical role pathologists will play in the evolution of computational pathology algorithms.


Local pH-dependent conformational changes leading to proteolytic susceptibility of cystatin C.
Cystatin C, a cysteine protease inhibitor, was subject to hydrolysis at two sites when complexed with papain and in the presence of excess papain. A pH-dependent cleavage at His-86 increases Asp-87 was observed, as well as a pH-independent one at Gly-4 increases Lys-5. His-86 increases Asp-87 hydrolysis increased with decreasing pH and was characterized kinetically. It could be described by a single ionization with pKa = 3.4 +/- 0.2 and (kcat./Km)max. = 1.4 (+/- 0.4) x 10(4) M-1.s-1 at I = 0.3 M. C.d. spectroscopy, also at I = 0.3 M, demonstrated a conformational change with pKa = 3.2 +/- 0.2, indicating that the pH-dependence of hydrolysis was due to a conformational change in cystatin C. At I = 0.15 M, the pKa of the conformational change observed by c.d. shifted to 4.1 +/- 0.1. This indicates that at physiological ionic strength of 0.15 M, a significant proportion of cystatin C complexed with protease would be in a proteolytically labile conformation over the pH range 4.5 to 5, which is encountered in lysosomes. This may constitute a mechanism for clearing inappropriately localized cystatins. A pH-dependent conformational variability in this region of the inhibitor could explain the differences in the X-ray crystallographic and n.m.r. structures of the homologous chicken cystatin. The ionic-strength dependence of ionization indicates a hydrophobic stabilization of the ionizable group. The lack of pH-dependence of hydrolysis at Gly-4 increases Lys-5, with kcat./Km = 220 +/- 41 M-1.s-1 in the pH range 3.89 to 7.96 was unexpected in light of the normal, bell-shaped pH-dependence of papain-catalysed hydrolyses. This may reflect a different rate-limiting step of cystatin C hydrolysis.


Enhancement of ricin cytotoxicity in Chinese hamster ovary cells by depletion of intracellular K+: evidence for an Na+/H+ exchange system in Chinese hamster ovary cells.
Depletion of intracellular K+ has been reported to result in an arrest of the formation of coated pits in human fibroblasts (Larkin, J.M., M.S. Brown, J.L. Goldstein, and R.G.W. Anderson, 1983, Cell, 33:273-285). We have studied the effects of K+ depletion on the cytotoxicities of ricin, Pseudomonas exotoxin A, and diphtheria toxin in Chinese hamster ovary (CHO) cells. The cytotoxicities of ricin and Pseudomonas toxin were enhanced in K+-depleted CHO cells whereas the cytotoxicity of diphtheria toxin was reduced by K+ depletion. The effects of NH4Cl on the cytotoxicities of ricin, Pseudomonas toxin, and diphtheria toxin were found to be similar to those of K+ depletion, and there were no additive or synergistic effects on ricin cytotoxicity by NH4Cl in K+-depleted medium. The enhancement of ricin cytotoxicity by K+ depletion could be completely reversed by the addition of K+, Rb+, and partially by the addition of Cs+, before the ricin treatment, whereas Li+ was ineffective. These protective effects of K+ or Rb+ requires a functional Na+/K+ ATPase. CHO cells grown in K+-depleted media were found to contain 6.3-fold increase in intracellular Na+ level, concomitant with a 10-fold reduction in intracellular K+ level. The enhanced cytotoxicity of ricin in K+-free medium and the increased uptake of Na+ could be abolished by amiloride or amiloride analogues, which are known to be potent inhibitors of the Na+/H+ antiport system. Our results suggest that a depletion of intracellular K+ results in an influx of Na+, which is accompanied by the extrusion of H+. Consequently, there is an alkalinization of the cytosol and the ricin-containing endosomes. As a result, ricin is more efficiently released from the endosomes in-K+-depleted cells. Results from the studies of the binding, internalization, and degradation of 125I-ricin, and the kinetics of inhibition of protein synthesis by ricin in K+-depleted cells are consistent with this working hypothesis.


Vitamin d deficiency and insufficiency in obese children and adolescents and its relationship with insulin resistance.
Objectives. We aimed to determine the relationship between insulin resistance and serum 25-hydroxyvitamin D (25-OHD) levels in obese children and their nonobese peers. Materials and Methods. Included in the study group were 188 obese children (aged 9-15 years), and 68 age- and gender-matched healthy children of normal weight as control group. Anthropomorphic data were collected on patients and fasting serum glucose, insulin, serum lipids, alanine aminotransaminase (ALT) and 25-OHD were measured. The homeostatic model assessment of insulin resistance (HOMA-IR) was calculated in both groups. Results. The levels of 25-OHD in the obese group were significantly lower than those of the nonobese (P = 0.002). HOMA-IR, triglycerides, low-density lipoprotein, and ALT levels in the obese group were significantly higher than values of control group (P < 0.001 and P = 0.002, resp.). In the obese group, vitamin D deficiency, insufficiency, and sufficiency (25-OHD < 10 ng/dl, < 20, >10 ng/dl; > 20 ng/dl, resp.) were not correlated with HOMA-IR (r : -0.008, P = 0.935). HOMA-IR was negatively correlated with BMI, BMI SDS, and BMI%, and triglycerides, low-density lipoprotein, and ALT levels (P < 0.001). Conclusion. The insulin resistance of the obese subjects who were vitamin D deficient and insufficient did not statistically differ from those with vitamin D sufficiency. Low 25-hydroxyvitamin D levels were not related with higher insulin resistance in obese children and adolescents. In obese subjects, insulin resistance was affected more from BMI, BMI SDS, and BMI% than from 25-hydroxyvitamin D levels.


Glyphosate-based herbicide exposure during pregnancy and lactation malprograms the male reproductive morphofunction in F1 offspring.
One of the most consumed pesticides in the world is glyphosate, the active ingredient in the herbicide ROUNDUP®. Studies demonstrate that glyphosate can act as an endocrine disruptor and that exposure to this substance at critical periods in the developmental period may program the fetus to induce reproductive damage in adulthood. Our hypothesis is that maternal exposure to glyphosate during pregnancy and lactation in mice will affect the development of male reproductive organs, impairing male fertility during adult life. Female mice consumed 0.5% glyphosate-ROUNDUP® in their drinking water [glyphosate-based herbicide (GBH) group] or filtered water [control (CTRL) group] from the fourth day of pregnancy until the end of the lactation period. Male F1 offspring were designated, according to their mother's treatment, as CTRL-F1 and GBH-F1. Female mice that drank glyphosate displayed reduced body weight (BW) gain during gestation, but no alterations in litter size. Although GBH male F1 offspring did not exhibit modifications in BW, they demonstrated delayed testicular descent. Furthermore, at PND150, GBH-F1 mice presented a lower number of spermatozoa in the cauda epididymis and reduced epithelial height of the seminiferous epithelium. Notably, intratesticular testosterone concentrations were enhanced in GBH-F1 mice; we show that it is an effect associated with increased plasma and pituitary concentrations of luteinizing hormone. Therefore, data indicate that maternal exposure to glyphosate-ROUNDUP® during pregnancy and lactation may lead to decreased spermatogenesis and disruptions in hypothalamus-pituitary-testicular axis regulation in F1 offspring.


Antigens for CD4 and CD8 T cells in tuberculosis.
Tuberculosis (TB), caused by infection with Mycobacterium tuberculosis (MTB), represents an important cause of morbidity and mortality worldwide for which an improved vaccine and immunodiagnostics are urgently needed. CD4(+) and CD8(+) T cells play an important role in host defense to TB. Definition of the antigens recognized by these T cells is critical for improved understanding of the immunobiology of TB and for development of vaccines and diagnostics. Herein, the antigens and epitopes recognized by classically HLA class I- and II-restricted CD4(+) and CD8(+) T cells in humans infected with MTB are reviewed. Immunodominant antigens and epitopes have been defined using approaches targeting particular TB proteins or classes of proteins and by genome-wide discovery approaches. Antigens and epitopes recognized by classically restricted CD4(+) and CD8(+) T cells show extensive breadth and diversity in MTB-infected humans.


Occupational medicine clinical practice data reveal increased injury rates among Hispanic workers.
Minnesota has an ethnically diverse labor force, with the largest number of refugees per capita in the United States. In recent years, Minnesota has been and continues to be a major site for immigrant and refugee resettlement in the United States, with a large population of both immigrant and native born Hmong, Hispanic, and East Africans. This study seeks to evaluate the injury risk among the evolving minority workforce in the Minnesota Twin Cities region. A retrospective cohort study identifying work-related injuries following pre-employment examinations was performed using electronic health records from a large multi-clinic occupational medicine practice. Preplacement examinations and subsequent work-related injuries were pulled from the electronic health record using representative ICD-10 codes for surveillance examinations and injuries. This study included patient records collected over a 2-year period from January 1, 2015, through December, 2016. The patients in this cohort worked in a wide-array of occupations including production, assembly, construction, law enforcement, among others. Hispanic minority workers were twice as likely to be injured at work compared with White workers. Hispanics were 2.89 times more likely to develop back injuries compared with non-Hispanic workers, and 1.86 times more likely to develop upper extremity injuries involving the hand, wrist, or elbow. Clinical practice data shows that Hispanic workers are at increased risk for work-related injuries in Minnesota. They were especially susceptible to back and upper extremity injuries. Lower injury rates in non-Hispanic minority workers, may be the result of injury underreporting and require further investigation.


IP-1: a dominant inhibitor of Fos/Jun whose activity is modulated by phosphorylation.
Transcription factor AP-1 is inducible by phorbol esters and thus could be considered to be one final target of the protein kinase C signal transduction pathway. AP-1 consists of the products of the fos and jun oncogenes, which associate as dimers to bind TPA-responsive promoter elements (TRE) efficiently. We show that AP-1 activity is modulated by an inhibitory protein (IP-1), present both in the nucleus and cytoplasm of several cell types. IP-1 specifically blocks DNA binding of AP-1 from nuclear extracts and of in vitro synthesized Fos/Jun proteins. It is a labile protein of 30-40 kd, which exerts its activity only in the nonphosphorylated form. Block of IP-1 function is obtained by PKA-mediated phosphorylation, possibly suggesting a cross talk mechanism at transcriptional level. Competition experiments with synthetic peptides suggest that IP-1 could interact with Fos and/or Jun leucine zippers. We speculate that IP-1 might act as a transcriptional antioncogene.


Injury of the external branch of the superior laryngeal nerve in thyroid surgery.
The external branch of the superior laryngeal nerve (EBSLN) innervates the cricothyroid muscle (CTM) to promote lengthening and thinning of the vocal fold, thus increasing voice pitch. The close relation with the superior thyroid vessels (STV) puts the EBSLN in risk every time the superior pole of the thyroid is dissected. It travels downward to innervate the CTM, lateral to the thyroid cartilage and to the inferior pharyngeal constrictor muscle (IPCM), being eventually covered by this muscle fibers as it approaches its entry point. During its descending course, the EBSLN curves and crosses the STV posteriorly. The lower this crossing occurs in the neck, the higher the risk of surgical damage to the nerve by transection, traction, entrapment, thermal damage or disrupted blood supply. The chances of surgical trauma are also increased by size and weight of the specimen, shorter neck length and non-white ethnicity. Voice changes following thyroid surgery are common and multifactorial. The actual rate of vocal impairment due to EBSLN injury is unclear, since changes to the everyday speaking voice can be minimal and laryngeal findings are usually subtle and controversial. CTM electroneuromyography (EMG) is the most accurate tool to diagnose abnormal EBSLN conductivity, but it is technically difficult and barely applicable in routine practice. Recommended approaches to prevent injury include: (I) individual distal ligature of the STV by the thyroid capsule; (II) visual identification of the nerve and its trajectory and (III) electrostimulation with either observation of CTM twitch or intraoperative nerve monitoring (IONM) via dedicated endotracheal tube electrodes. There is accumulating evidence that a combination of visual and standardized electrophysiological EBSLN identification with meticulous division of the STV improves preservation rates. IONM bears the additional benefits of prognostication, quantification and documentation of neural function once it allows intraoperative laryngeal EMG.


MacIntosh arthroplasty for the rheumatoid knee: a 10-year follow up.
The results of 75 MacIntosh arthroplasties performed for rheumatoid arthritis of the knee in 63 patients were reviewed at least 10 years after surgery. Forty-two knees in 35 patients were available for assessment. Eleven arthroplasties had been revised to total knee replacement without difficulty because of pain or poor function. The remaining 31 knees in 25 patients gave good or excellent results in 22 cases, fair in eight, and poor in one. Seven patients could not be traced, and 21 patients representing 26 knees had died. At least half these knees had given satisfactory results immediately before death judged by review of the case notes. The difficulty of comparing functional status with the preoperative state because of progressive multiarticular disease was highlighted. Although greater angular deformities preoperatively reduced the chance of success in the medium term, late failure of the arthroplasty after five years was very rare. Approximately two-thirds of all the arthroplasties performed gave satisfactory results at 10-year follow up or until the time of death.


Thrombosis and drug-eluting stents: an objective appraisal.
Stent thrombosis (ST) after percutaneous coronary intervention has been the focus of intense interest because of its attendant morbidity and mortality. There is controversy about several facets of the problem. These include the frequency of ST with drug-eluting stents (DES) versus bare-metal stents (BMS), the timing of the event, clinical consequences, risk factors, adjunctive therapy, and new preventive approaches. Information has accrued rapidly from several sources, including randomized controlled clinical trials of DES versus BMS in carefully selected subsets of patients and registry experiences in larger patient groups, which provide a more universal real-world picture. The results from these different data sets are not completely concordant. However, several general conclusions can be made: 1) ST is an infrequent but very severe complication of both BMS and DES; 2) at the present time, during 4 years of follow-up from randomized controlled trials that compared DES and BMS, there is no apparent difference in overall ST frequency, although the time course for occurrence appears to differ, with a relative numeric excess of ST late after DES implant; 3) despite this relative imbalance, no differences in the end points of death or death and infarction between DES and BMS are observed; 4) longer-term follow-up of these patients as well as larger angiographic and clinical subsets of patients who receive this technology outside of randomized trials are required to fully study this issue; and 5) advances in stent platforms for drug elution as well as adjunctive pharmacologic therapy are being evaluated to enhance long-term safety.


Arrests for child pornography production: data at two time points from a national sample of U.S. law enforcement agencies.
This study collected information on arrests for child pornography (CP) production at two points (2000-2001 and 2006) from a national sample of more than 2,500 law enforcement agencies. In addition to providing descriptive data about an understudied crime, the authors examined whether trends in arrests suggested increasing CP production, shifts in victim populations, and challenges to law enforcement. Arrests for CP production more than doubled from an estimated 402 in 2000-2001 to an estimated 859 in 2006. Findings suggest the increase was related to increased law enforcement activity rather than to growth in the population of CP producers. Adolescent victims increased, but there was no increase in the proportion of arrest cases involving very young victims or violent images. Producers distributed images in 23% of arrest cases, a proportion that did not change over time. This suggests that much CP production may be primarily for private use. Proactive law enforcement operations increased, as did other features consistent with a robust law enforcement response.


A cross selectional survey in a critical care: the job satisfaction and functioning team of the health professionals.
Health care workers, especially those who are part of the OS core, are essential in the delivery of services, as they represent the institution at the time of the contact with the user and they represent also the image of the organization. Health administrations, therefore, are called to improve the performance through a better motivation and satisfaction of the staff, in view of two strategic aspects: job satisfaction of professionals and team collaboration. Between January and September 2014, a survey at the OU (UUOO) intensive care and sub intensive has been made inside three hospitals in Emilia Romagna. It' s been a multicentre cross-sectional quantitative study by administering a self-report questionnaire designed to investigate the different constructs. On 742 questionnaires were spread 454 professionals gave it back (response rate = 73%). Of those, 273 (60.1%) were nurses, 119 (26.2%) were physicians and 62 (13.7%) were healthcare operators. 62 (13.7%) Job Satisfaction was measured with the McCloskey Mueller Satisfaction Scale. Team Functioning was measured with the Index of Interprofessional Team Collaboration. Results from MANOVA indicated that physicians were less satisfied of scheduling than both nurses and healthcare operators. For professional opportunities, instead, healthcare operators showed the lower level of satisfaction. The participants seem to perceive a high level of team effectiveness and therefore the professionals involved in the care of critically ill patients than the two dimensions analysed, (reflection between the processes and interdependent roles), also state a greater tendency to respect the roles interdisciplinary, maintaining their professional autonomy and a lower tendency to use critical thinking to act professionally in order to improve the effectiveness of care provided. The study results oriented healthcare administrators, to take paths that feed the job satisfaction and the collaboration of professionals by developing the aspects investigated. Considering the shared perception among the professions studied, compared to the constructs under investigation, it seems to be clear how the routes should be designed in a systematic order to involve, in an integrated way the best professionals (nurses, doctors and OSS) involved in taking managing critical patients. No statistically significant difference have been found between these three professions considered, on most dimensions of job satisfaction investigated (relations between colleagues, 'social interactions, work-family balance, time work organization and working professional opportunity). There is a difference between two specific factors: the work time planning organization and wich is seen less by the nurses, while the job opportunities that are perceived less from the OOS.


Structure of the diaminopimelate epimerase DapF from Mycobacterium tuberculosis.
The meso (or D,L) isomer of diaminopimelic acid (DAP), a precursor of L-lysine, is a key component of the pentapeptide linker in bacterial peptidoglycan. While the peptidoglycan incorporated in the highly complex cell wall of the pathogen Mycobacterium tuberculosis structurally resembles that of Escherichia coli, it is unique in that it can contain penicillin-resistant meso-DAP-->meso-DAP linkages. The interconversion of L,L-DAP and meso-DAP is catalysed by the DAP epimerase DapF, a gene product that is essential in M. tuberculosis. Here, the crystal structure of the ligand-free form of M. tuberculosis DapF (MtDapF) refined to a resolution of 2.6 A is reported. MtDapF shows small if distinct deviations in secondary structure from the two-domain alpha/beta-fold of the known structures of Haemophilus influenzae DapF and Bacillus anthracis DapF, which are in line with its low sequence identity (<or=27%) to the former. Modelling the present structure onto that of L,L-aziridino-DAP-bound H. influenzae DapF illustrates that a rigid-body movement of domain II and a rearrangement of the B4-A2 loop (residues 80-90) of domain I are likely to accompany the transition from the present inactive form to a catalytically competent enzyme. Despite a highly conserved active-site architecture, the model indicates that stabilization of the DAP backbone occurs in MtDapF through a tyrosine residue that is specific to mycobacterial DAP epimerases.


Elderly parent health and the migration decisions of adult children: evidence from rural China.
Recent research has shown that participation in migrant labor markets has led to substantial increases in income for families in rural China. This article addresses the question of how participation is affected by elderly parent health. We find that younger adults are less likely to work as migrants when a parent is ill. Poor health of an elderly parent has less impact on the probability of employment as a migrant when an adult child has siblings who may be available to provide care. We also highlight the potential importance of including information on nonresident family members when studying how parent illness and elder care requirements influence the labor supply decisions of adult children.


Chemiluminescence sequential injection immunoassay for vitellogenin using magnetic microbeads.
A rapid and sensitive immunoassay for the determination of carp vitellogenin (Vg) is described. The method involves a sequential injection analysis (SIA) system equipped with a chemiluminescence detector and a samarium-cobalt magnet. An anti-Vg monoclonal antibody, immobilized on magnetic beads, was used as a solid support for the immunoassay. The introduction, trapping and release of the magnetic beads in the flow cell were controlled by a samarium-cobalt magnet and the flow of the carrier solution. The immunoassay was based on a sandwich immunoreaction of anti-Vg monoclonal antibody (primary antibody) on the magnetic beads, Vg, and the anti-Vg antibody labeled with horseradish peroxidase (HRP) (secondary antibody), and was based on a subsequent chemiluminescence reaction of HRP with hydrogen peroxide and p-iodophenol, in a luminol solution. The magnetic beads to which the primary antibody was immobilized were prepared by coupling the primary antibody with the magnetic beads after an agarose-layer on the surface of the magnetic beads was epoxidized. The primary antibody-immobilized magnetic beads were introduced, and trapped in the flow cell equipped with the samarium-cobalt magnet, a Vg sample solution, an HRP-labeled secondary antibody solution and the luminol solution were sequentially introduced into the flow cell based on an SIA programmed sequence. Chemiluminescence emission was monitored by means of a photomultiplier located at the upper side of the flow cell. The optimal incubation times both for the first and second immunoreactions were determined to be 20min. A concave calibration curve was obtained between Vg concentration and chemiluminescence intensity when various concentrations of standard Vg samples (2-100ngmL(-1)) were applied to the SIA system under optimal conditions. In spite of a narrow working range, the lower detection limit of the immunoassay was about 2ngmL(-1).


Survey-based socio-economic data from slums in Bangalore, India.
In 2010, an estimated 860 million people were living in slums worldwide, with around 60 million added to the slum population between 2000 and 2010. In 2011, 200 million people in urban Indian households were considered to live in slums. In order to address and create slum development programmes and poverty alleviation methods, it is necessary to understand the needs of these communities. Therefore, we require data with high granularity in the Indian context. Unfortunately, there is a paucity of highly granular data at the level of individual slums. We collected the data presented in this paper in partnership with the slum dwellers in order to overcome the challenges such as validity and efficacy of self reported data. Our survey of Bangalore covered 36 slums across the city. The slums were chosen based on stratification criteria, which included geographical location of the slum, whether the slum was resettled or rehabilitated, notification status of the slum, the size of the slum and the religious profile. This paper describes the relational model of the slum dataset, the variables in the dataset, the variables constructed for analysis and the issues identified with the dataset. The data collected includes around 267,894 data points spread over 242 questions for 1,107 households. The dataset can facilitate interdisciplinary research on spatial and temporal dynamics of urban poverty and well-being in the context of rapid urbanization of cities in developing countries.


The effect of higher-order dispersion on slow light propagation in photonic crystal waveguides.
We have studied the dispersion of ultrafast pulses in a photonic crystal waveguide as a function of optical frequency, in both experiment and theory. With phase-sensitive and time-resolved near-field microscopy, the light was probed inside the waveguide in a non-invasive manner. The effect of dispersion on the shape of the pulses was determined. As the optical frequency decreased, the group velocity decreased. Simultaneously, the measured pulses were broadened during propagation, due to an increase in group velocity dispersion. On top of that, the pulses exhibited a strong asymmetric distortion as the propagation distance increased. The asymmetry increased as the group velocity decreased. The asymmetry of the pulses is caused by a strong increase of higher order dispersion. As the group velocity was reduced to 0.116(9) .c, we found group velocity dispersion of -1.1(3) .10(6) ps(2)/km and third order dispersion of up to 1.1(4) .10(5) ps(3)/km. We have modelled our interferometric measurements and included the full dispersion of the photonic crystal waveguide. Our mathematical model and the experimental findings showed a good correspondence. Our findings show that if the most commonly used slow light regime in photonic crystals is to be exploited, great care has to be taken about higher-order dispersion.


Nutrition and physical activity intervention for families with familial hypercholesterolaemia: protocol for a pilot randomised controlled feasibility study.
Untreated heterozygous familial hypercholesterolaemia (FH) causes high low-density lipoprotein cholesterol (LDL-C) levels and increased cardiovascular disease (CVD) risk. Despite pharmacological treatment, many treated individuals remain at higher CVD risk than non-affected individuals. This may be due to LDL-C targets not being met and presence of other CVD risk factors. Adhering to dietary and physical activity (PA) recommendations developed for individuals with FH may further reduce CVD risk. However, there is insufficient research to support the efficacy of adhering to these guidelines on LDL-C and other CVD risk factors. The need for studies to investigate the effectiveness of nutrition and PA interventions in the FH population has been widely recognised and recommended. This paper describes the protocol of a pilot, randomised controlled trial designed to evaluate the feasibility and acceptability of a specifically developed nutrition and PA intervention aimed at improving the dietary intakes and PA levels of families with FH. A two-arm randomised waitlist-controlled pilot trial will be conducted across three National Health Service (NHS) sites in England, UK. Twenty-four young people with FH, aged 10-18 years, and their affected parent, will be recruited and randomly assigned to the intervention or waitlist and usual care control. The primary aim is to provide evidence for the feasibility and acceptability of delivering the intervention, explored quantitatively (rates of recruitment, retention and outcome measure completeness) and qualitatively (qualitative interviews). The secondary aim is to provide evidence for the potential efficacy of the intervention on dietary intake, PA, sedentary time, body composition, CVD risk factors and quality of life determined at baseline and endpoint assessments. The intervention will involve an hour-long consultation with a dietitian at baseline and four follow-up contacts across the 12-week intervention. It has been specifically developed for use with individuals with FH and incorporates behavioural change techniques to target identified enablers and barriers to adherence in this population. This trial will estimate the feasibility and acceptability of the nutrition and PA intervention delivered to young people and parents with FH. If appropriate, this study can be used to inform the design of an adequately powered definitive trial. ISRCTN, ISRCTN24880714. Registered 07/06/2018, http://www.isrctn.com/ISRCTN24880714.


An unusual H2 sorption mechanism in PCN-14: insights from molecular simulation.
Simulations of H2 sorption were performed in PCN-14, a metal-organic framework (MOF) that consists of Cu(2+) ions coordinated to 5,5'-(9,10-anthracenediyl)diisophthalate (adip) linkers. This MOF displays an excess H2 uptake of 2.70 wt% at 77 K and 1.0 atm and an initial H2Qst value of 8.6 kJ mol(-1) according to previous experimental measurements. The experimental H2 sorption isotherms and Qst values in PCN-14 were reproduced in simulations using well-known H2 potentials that have been widely used for MOF-H2 theoretical studies. H2 sorption in PCN-14 was dominated by repulsion/dispersion energetics; this allowed the experimental observables to be reproduced by a model that includes only Lennard-Jones parameters. The most energetically favorable H2 sorption site in PCN-14 corresponds to sorption within a small cage that is enclosed by three [Cu2(O2CR)4] units and three adip linkers. The anthracenyl rings of the adip linkers represent the secondary sorption sites within the MOF. In contrast to expectations, sorption of H2 onto the Cu(2+) ions of the copper paddlewheels was not observed within the simulations at low loading. The simulations revealed that the open-metal sites in PCN-14 were occupied at high loading. Control simulations of H2 sorption in PCN-14 for different cases in which the partial positive charge of one of the paddlewheel Cu(2+) ions was increased relative to the other revealed that sorption onto the open-metal sites can be captured if there is a very high positive charge on the metal. Otherwise, the calculated partial charge for the Cu(2+) ions in PCN-14 in this work was not high enough in magnitude to facilitate strong H2-metal interactions in simulation. This study shows the power of using molecular simulations to elucidate an unusual H2 sorption behavior in a MOF.


Comparison of Different Matrices as Potential Quality Control Samples for Neurochemical Dementia Diagnostics.
Assay-vendor independent quality control (QC) samples for neurochemical dementia diagnostics (NDD) biomarkers are so far commercially unavailable. This requires that NDD laboratories prepare their own QC samples, for example by pooling leftover cerebrospinal fluid (CSF) samples. To prepare and test alternative matrices for QC samples that could facilitate intra- and inter-laboratory QC of the NDD biomarkers. Three matrices were validated in this study: (A) human pooled CSF, (B) Aβ peptides spiked into human prediluted plasma, and (C) Aβ peptides spiked into solution of bovine serum albumin in phosphate-buffered saline. All matrices were tested also after supplementation with an antibacterial agent (sodium azide). We analyzed short- and long-term stability of the biomarkers with ELISA and chemiluminescence (Fujirebio Europe, MSD, IBL International), and performed an inter-laboratory variability study. NDD biomarkers turned out to be stable in almost all samples stored at the tested conditions for up to 14 days as well as in samples stored deep-frozen (at - 80°C) for up to one year. Sodium azide did not influence biomarker stability. Inter-center variability of the samples sent at room temperature (pooled CSF, freeze-dried CSF, and four artificial matrices) was comparable to the results obtained on deep-frozen samples in other large-scale projects. Our results suggest that it is possible to replace self-made, CSF-based QC samples with large-scale volumes of QC materials prepared with artificial peptides and matrices. This would greatly facilitate intra- and inter-laboratory QC schedules for NDD measurements.


On the Enantioselective HPLC Separation Ability of Sub-2 µm Columns: Chiralpak® IG-U and ID-U.
Silica with a particle size of 3-5 µm has been widely used as selector backbone material in 10-25 cm HPLC chiral columns. Yet, with the availability of 1.6 µm particles, shorter, high-efficiency columns practical for minute chiral separations are possible to fabricate. Herein, we investigate the use of two recently commercialized sub-2 µm columns with different substituents. Thus, Chiralpak® IG-U and ID-U were used in HPLC for the fast enantioseparation of a set of drugs. Chiralpak® IG-U [amylose tris (3-chloro-5-methylphenylcarbamate)] has two substituents on the phenyl ring, namely, a withdrawing chlorine group in the third position and a donating group in the fifth position. Chiralpak® ID-U [amylose tris (3-chlorophenylcarbamate)] has only one substituent on the phenyl ring, namely a withdrawing chlorine group. Their applications in three liquid chromatography modes, namely, normal phase, polar organic mode, and reversed phase, were demonstrated. Both columns have similar column parameters (50 mm length, 3 mm internal diameter, and 1.6 µm particle size) with the chiral stationary phase as the only variable. Improved chromatographic enantioresolution was obtained with Chiralpak® ID-U. Amino acids partially separated were reported for the first time under an amylose-based sub-2-micron column.


Associations between comorbidities and immediate and one-year outcomes following supervised exercise therapy and patient education - A cohort study of 24,513 individuals with knee or hip osteoarthritis.
To investigate if comorbidities are associated with change in health outcomes following an 8-week exercise and education program in knee and hip osteoarthritis (OA). We included 24,513 individuals with knee or hip OA from the Good Life with osteoArthritis in Denmark (GLA:D®). GLA:D® consists of two patient education sessions and 12 supervised exercise sessions. Before the program, individuals self-reported having one or more of 11 common comorbidities. Physical function was assessed using the 40-m Fast-Paced Walk Test (FPWT, m/sec) before and immediately after the program. Pain intensity and health-related quality of life was self-reported before, immediately after, and at 12 months post-intervention using a visual analogue scale (VAS, 0-100) and the EQ-5D-5L index (-0.624 to 1.000), respectively. Associations of comorbidity combinations with change in outcomes immediately and at 12 months was estimated using mixed linear regression. Individuals with OA improved on average 0.12 m/s (95%CI 0.12 to 0.13) in 40-m FPWT, -12.7 mm (95%CI -13.2 to -12.2) in VAS, and 0.039 (95%CI 0.036 to 0.041) in EQ-5D-5L from before to immediately after the intervention with minor additional improvements at 12 months. Despite that individuals with comorbidities had worse baseline scores in all outcomes than individuals without comorbidities, they had similar levels of improvement immediately and 12 months after the intervention. Comorbidities are not associated with worse nor better health outcomes following an 8-week exercise and education program in individuals with OA, suggesting exercise as a viable treatment option for individuals with OA, irrespective of comorbidities.


Water deprivation test in children with polyuria.
Polyuria is an uncommon clinical presentation in paediatric practice. When diabetes mellitus has been excluded by history taking and preliminary investigations and impaired renal concentrating ability is confirmed, water deprivation test (WDT) is necessary to differentiate among central diabetes insipidus (CDI), nephrogenic diabetes insipidus, or primary polydipsia. Traditionally, responsiveness to desmopressin injection is defined as urine osmolality >750 mOsm/kg. However, that level could not be reached in the review of our patients. We discuss how to arrive at the diagnosis of CDI in WDT. An approach to polyuria and WDT will also be discussed.


The pericellular matrix as a transducer of biomechanical and biochemical signals in articular cartilage.
The pericellular matrix (PCM) is a narrow tissue region surrounding chondrocytes in articular cartilage, which together with the enclosed cell(s) has been termed the "chondron." While the function of this region is not fully understood, it is hypothesized to have important biological and biomechanical functions. In this article, we review a number of studies that have investigated the structure, composition, mechanical properties, and biomechanical role of the chondrocyte PCM. This region has been shown to be rich in proteoglycans (e.g., aggrecan, hyaluronan, and decorin), collagen (types II, VI, and IX), and fibronectin, but is defined primarily by the presence of type VI collagen as compared to the extracellular matrix (ECM). Direct measures of PCM properties via micropipette aspiration of isolated chondrons have shown that the PCM has distinct mechanical properties as compared to the cell or ECM. A number of theoretical and experimental studies suggest that the PCM plays an important role in regulating the microenvironment of the chondrocyte. Parametric studies of cell-matrix interactions suggest that the presence of the PCM significantly affects the micromechanical environment of the chondrocyte in a zone-dependent manner. These findings provide support for a potential biomechanical function of the chondrocyte PCM, and furthermore, suggest that changes in the PCM and ECM properties that occur with osteoarthritis may significantly alter the stress-strain and fluid environments of the chondrocytes. An improved understanding of the structure and function of the PCM may provide new insights into the mechanisms that regulate chondrocyte physiology in health and disease.


A socioeconomic deprivation index for small areas in Denmark.
To describe the development of a deprivation index for Danish parishes and to investigate its association with all-cause mortality compared with the Townsend index and individual-level factors. Nine socioeconomic factors were aggregated to the parish level from individual-level register data comprising the entire Danish population in 2005. A principal component analysis was conducted to reduce the number of factors and to apply weights. An ecological analysis investigated the association between the Danish Deprivation Index (DANDEX) and standardised mortality ratios in Danish parishes. Results were compared with the Townsend index and a possible modifying effect of population density was investigated. Sensitivity of the index was investigated with multilevel survival analyses evaluating the association between all-cause mortality and DANDEX, the Townsend Index, individual-level socioeconomic factors, and population density. DANDEX consists of two components measuring socioeconomic properties of all 2119 Danish parishes. The first component measures deprivation related to housing (house and car ownership) while the second component is related to classical measures of socioeconomic status (education, income, occupation). A clear gradient in standardised mortality ratios was seen across quintiles of both index components. Modifying effects were seen when stratifying the index components by population density. In a frailty model, DANDEX accounted for 76% of the between-parish variation in all-cause mortality, while the Townsend index and individual socioeconomic factors accounted for 71 and 76%, respectively. The index can be used to identify Danish parishes by their levels of deprivation and it provides municipalities with a tool to allocate resources to the geographic areas where they are most needed.


Are small-scale irrigators water use efficient? Evidence from lake Naivasha basin, Kenya.
With increasing water scarcity and competing uses and users, water use efficiency is becoming increasingly important in many parts of developing countries. The lake Naivasha basin has an array of different water users and uses ranging from large scale export market agriculture, urban domestic water users to small holder farmers. The small scale farmers are located in the upper catchment areas and form the bulk of the users in terms of area and population. This study used farm household data to explore the overall technical efficiency, irrigation water use efficiency and establish the factors influencing water use efficiency among small scale farmers in the Lake Naivasha basin in Kenya. Data envelopment analysis, general algebraic and modeling system, and Tobit regression methods were used in analyzing cross sectional data from a sample of 201 small scale irrigation farmers in the lake Naivasha basin. The results showed that on average, the farmers achieved only 63 % technical efficiency and 31 % water use efficiency. This revealed that substantial inefficiencies occurred in farming operations among the sampled farmers. To improve water use efficiency, the study recommends that more emphasis be put on orienting farmers toward appropriate choice of irrigation technologies, appropriate choice of crop combinations in their farms, and the attainment of desirable levels of farm fragmentation.


Osteoarthritis-like pathologic changes in the knee joint induced by environmental disruption of circadian rhythms is potentiated by a high-fat diet.
A variety of environmental factors contribute to progressive development of osteoarthritis (OA). Environmental factors that upset circadian rhythms have been linked to various diseases. Our recent work establishes chronic environmental circadian disruption - analogous to rotating shiftwork-associated disruption of circadian rhythms in humans - as a novel risk factor for the development of OA. Evidence suggests shift workers are prone to obesity and also show altered eating habits (i.e., increased preference for high-fat containing food). In the present study, we investigated the impact of chronic circadian rhythm disruption in combination with a high-fat diet (HFD) on progression of OA in a mouse model. Our study demonstrates that when mice with chronically circadian rhythms were fed a HFD, there was a significant proteoglycan (PG) loss and fibrillation in knee joint as well as increased activation of the expression of the catabolic mediators involved in cartilage homeostasis. Our results, for the first time, provide the evidence that environmental disruption of circadian rhythms plus HFD potentiate OA-like pathological changes in the mouse joints. Thus, our findings may open new perspectives on the interactions of chronic circadian rhythms disruption with diet in the development of OA and may have potential clinical implications.


Bone marrow adipogenic lineage precursors promote osteoclastogenesis in bone remodeling and pathologic bone loss.
Bone is maintained by coupled activities of bone-forming osteoblasts/osteocytes and bone-resorbing osteoclasts. Alterations in this relationship can lead to pathologic bone loss such as osteoporosis. It is well known that osteogenic cells support osteoclastogenesis via production of RANKL. Interestingly, our recently identified bone marrow mesenchymal cell population-marrow adipogenic lineage precursors (MALPs) that form a multidimensional cell network in bone-was computationally demonstrated to be the most interactive with monocyte-macrophage lineage cells through high and specific expression of several osteoclast regulatory factors, including RANKL. Using an adipocyte-specific Adipoq-Cre to label MALPs, we demonstrated that mice with RANKL deficiency in MALPs have a drastic increase in trabecular bone mass in long bones and vertebrae starting from 1 month of age, while their cortical bone appears normal. This phenotype was accompanied by diminished osteoclast number and attenuated bone formation at the trabecular bone surface. Reduced RANKL signaling in calvarial MALPs abolished osteolytic lesions after LPS injections. Furthermore, in ovariectomized mice, elevated bone resorption was partially attenuated by RANKL deficiency in MALPs. In summary, our studies identified MALPs as a critical player in controlling bone remodeling during normal bone metabolism and pathological bone loss in a RANKL-dependent fashion.


Adrenocortical and behavioral responses to limb restraint in human neonates.
Two experiments were conducted on human neonates examining their behavior and adrenocortical activity in response to physical restraint. In both experiments, newborns (9 per condition in Experiment I; 10 per condition in Experiment II) were tested in one of three conditions: Restraint, which involved strapping newborns to a circumstraint board for 20 min; Strapping Control, which involved strapping newborns to the board and then immediately taking them off the board and returning them to their cribs; and Control, which did not involve any restraining manipulation. In Experiment I, blood samples for plasma cortisol determination were obtained via heel-stick immediately prior to the assigned manipulations and 30 min later. In Experiment II, only the postmanipulation blood sample was taken. Two behavioral states, Active Awake and Crying, were examined as indices of behavioral distress. In Experiment I, the effects of the blood sampling procedure on behavioral distress and plasma cortisol were significant; however, no significant effects of restraint were observed. In Experiment II, restraint was associated with small but significant increases in behavioral distress and in plasma cortisol levels.


A global database for metacommunity ecology, integrating species, traits, environment and space.
The use of functional information in the form of species traits plays an important role in explaining biodiversity patterns and responses to environmental changes. Although relationships between species composition, their traits, and the environment have been extensively studied on a case-by-case basis, results are variable, and it remains unclear how generalizable these relationships are across ecosystems, taxa and spatial scales. To address this gap, we collated 80 datasets from trait-based studies into a global database for metaCommunity Ecology: Species, Traits, Environment and Space; "CESTES". Each dataset includes four matrices: species community abundances or presences/absences across multiple sites, species trait information, environmental variables and spatial coordinates of the sampling sites. The CESTES database is a live database: it will be maintained and expanded in the future as new datasets become available. By its harmonized structure, and the diversity of ecosystem types, taxonomic groups, and spatial scales it covers, the CESTES database provides an important opportunity for synthetic trait-based research in community ecology.


Klotho Protein Protects Human Keratinocytes from UVB-Induced Damage Possibly by Reducing Expression and Nuclear Translocation of NF-κB.
BACKGROUND UV-related skin disease such as actinic keratosis is a major concern in public health. In view of the cell injury induced by UVB, Klotho protein it is an ideal therapy to eliminate UVB-induced cell damages and the associated signaling pathways. MATERIAL AND METHODS To gain insights into the potential role of Klotho and the underlying molecular mechanism, we constructed a Klotho-overexpress HaCaT cell line and assessed the protection against UVB insults. The effects of exposure to UVB radiation on the human keratinocyte HaCaT cells, including cell growth, apoptosis, and changes of selected biomarkers, were measured by CCK-8, flow cytometry, Quantitative real-time PCR, and Western blot analysis. RESULTS We found that enhanced NF-κB activity was accompanied by decreased expression of the anti-aging protein Klotho upon UVB stimulation, which was further confirmed with in vivo experiments. Overexpression of Klotho was able to considerably alleviate the UVB-induced damages to cells and reversed the UVB-caused biomarker changes to a great extent, which was comparable to the effects of administration of NF-κB inhibitor PDTC, suggesting the inhibition of nuclear translocation and DNA-binding activity of NF-κB. Furthermore, Klotho overexpression was proved to decrease the nuclear expression of NF-κB as much as the treatment with PDTC, which provides support for the direct regulation of NF-κB by Klotho. CONCLUSIONS Collectively, our work provides new insight into the potential role of Klotho in the context of UVB-induced injuries in human keratinocytes, as well as providing the basis for future study of new therapies against UV-related skin disease.


A highly selective naphthalimide-based ratiometric fluorescent probe for the recognition of tyrosinase and cellular imaging.
Tyrosinase is polyphenolic oxidase enzyme associated with the progression of various diseases. Therefore, for the recognition of tyrosinase, naphthalimide-based ratiometric fluorescent sensor probe was designed and synthesized. 3-Hydroxyphenyl, as the substrate unit for the enzyme, is an important feature of this design, which avoids the interference of other bio-analytes for the recognition of tyrosinase. When the sensor probe was excited at 425 nm, an intense blue emission band emerged at 467 nm. However, upon the addition of tyrosinase to the probe solution, the monophenolic unit oxidized to o-dihydroxy and consequently released the 4-aminonaphthalimide unit. As the oxidation reaction proceeded, the fluorescence emission at 535 nm started to increase gradually with an increase in the concentration of enzyme. Therefore, the sensor probe gives the ratiometric changes via fluorescence spectroscopy. The probe affords high selectivity and sensitivity to tyrosinase with a detection limit of 0.2 U mL-1. Furthermore, live cell images were recorded to assay the endogenous enzyme in A375 cells, which also show a dual color change in the presence of the L3 probe.


[Effect of propolis on insulin resistance in fructose-drinking rats].
Propolis, a honeybee product, contains a variety of biologically active substances. The present study was designed to investigate the effects of propolis on insulin resistance induced by fructose-drinking rats (FDR; type 2 diabetic animal model). Male Wistar rats (6 weeks old) received 15% fructose solution in drinking water for 8 weeks. FDR showed significant increases in plasma levels of insulin, Homeostasis Model Assessment ratio (HOMA-R, an index of insulin resistance), body weight, and systolic blood pressure but not blood glucose levels, when compared with control rats. Brazilian propolis extract (100 and 300 mg/kg, p.o.) treatment for 8 weeks significantly decreased the plasma level of insulin, HOMA-R, and body weight, increased plasma triglyceride levels without affecting blood glucose and total cholesterol levels, and tended to decrease systolic blood pressure. In isolated and perfused mesenteric vascular beds of FDR, propolis treatment resulted in a significant reduction of sympathetic nerve-mediated vasoconstrictor response to periarterial nerve stimulation (PNS; 8 Hz) and tended to increase the calcitonin gene-related peptide (CGRP) nerve-mediated vasodilator response to PNS, compared with those in untreated FDR. However, propolis treatment did not significantly affect norepinephrine-induced vasoconstriction and CGRP-induced vasodilation. These results suggest that propolis could be an effective functional food to prevent the development of insulin resistance.


Abnormal iodoprotein distribution and resistance to proteolysis in Gunn rat black thyroid. An ultrastructural and biochemical study.
Gunn rats have a marked deficiency in hepatic UDP-glucuronosyl transferase activity which results in hyperthyroxinemia and hyperbilirubinemia. Their thyroids show a brownish-black discoloration associated ultrastructurally with intracellular dense granules and intraluminal dense masses. In order to determine whether colloid composition and colloid proteolysis are altered in the thyroid of the Gunn rat compared with the Wistar rat, we studied the in situ resistance of thyroid proteins to in vitro proteolysis, the pattern of in vivo (125I) labeled thyroid iodoproteins and the proteolysis of isolated iodoprotein fractions in both strains of rats. For the cytochemical study, thin sections of aldehyde-fixed and plastic-embedded thyroid tissue were treated with 0.3 or 1% pronase in aqueous solution. With the low concentration of pronase, the secretory granules in C-cells and the apical vesicles in follicular cells were extensively digested in both strains of rats, whereas the colloid in the follicular lumen and the colloid droplets were only partially digested. With the high concentration of pronase, the colloid in the lumen and the colloid droplets were more markedly digested in both strains. In the presence of both concentrations of pronase, the dense granules and intraluminal dense masses were unchanged in the Gunn rats. The (125I) iodoprotein pattern was investigated 24 h after a single injection of (125I) iodide and by labeling at the isotopic equilibrium. It was found that the (125I) thyroglobulin fraction was reduced, whereas the (125I) 3-8 S fraction was increased in Gunn rats compared to Wistar rats. Pronase hydrolysis of the soluble (125I) iodine fraction showed similar pronase-resistant fractions in both strains with the single labeling procedure. At the isotopic equilibrium, the pronase resistant fraction was significantly increased in Gunn rats (Gunn 24.0 +/- 5.3%; Wistar: 13.7 +/- 3.1% of the soluble 125I) and a linear correlation was observed between the (125I) 3-8 S fraction of the soluble extract and the pronase-resistant fraction. These data suggest that iodocompounds of small molecular size and low turnover accumulate in the thyroid of the Gunn rat due to their strong resistance to in vivo hydrolysis. A local accumulation of 3-8 S iodocompounds may occur within the intracellular dense granules and intraluminal dense masses in the thyroid of Gunn rat.


Amsacta moorei entomopoxvirus expresses an active superoxide dismutase.
The entomopoxvirus from Amsacta moorei serves as the prototype of the group B entomopoxviruses. One of the interesting genes found in Amsacta moorei entomopoxvirus (AmEPV) is a superoxide dismutase (sod) (open reading frame AMV255). Superoxide dismutases (SODs) catalyze the conversion of superoxide radicals to hydrogen peroxide and oxygen. Many vertebrate poxviruses contain a sod gene, but to date, none have been demonstrated to be active. There are three families of SODs, characterized by their metal ion-binding partners, Fe, Mn, or Cu and Zn. Poxvirus enzymes belong to the Cu-Zn SOD family. Unlike inactive vertebrate poxvirus SODs, AMVSOD contains all the amino acids necessary for function. We expressed and purified a 6X-His-tagged version of the AMVSOD in Escherichia coli. The recombinant AMVSOD demonstrates superoxide dismutase activity both in an in situ gel assay and by stopped flow spectrophotometry. The k(cat)/K(m) for AMVSOD is 4 x 10(7) M(-1)s(-1). In infected cells, the AMVSOD protein behaves as a dimer and is catalytically active; however, disruption of the gene in AMEPV has little or no effect on growth of the virus in cell culture. An analysis of mRNA expression indicates that AMVsod is expressed late during infection of Lymantria dispar (Ld652) cells and produces a discrete nonpolydisperse transcript. Characterization of protein expression with a monoclonal antibody generated against AMVSOD confirms that the AMVSOD protein can be classified as a late, postreplicative gene. Therefore, AMVSOD is the first example of an active poxvirus SOD.


Reflexes elicited by acute stretch of atrial vs. pulmonary receptors in conscious dogs.
We measured hemodynamics and renal function in conscious dogs while partially obstructing blood flow at various sites within the thorax. Inflation of a balloon in the left atrium increased left atrial pressure (LAP) by 9 mmHg and caused a parallel increase in pulmonary arterial pressure (PAP); heart rate, arterial pressure, and total peripheral resistance increased; stroke volume and right atrial pressure decreased; and cardiac output remained unchanged. The increase in LAP was accompanied by a fourfold increase in urine flow and a threefold increase in sodium excretion. Plasma vasopressin (AVP) and renin activity (PRA) decreased. On the other hand, partial occlusion of the pulmonary veins or the main pulmonary artery produced similar increases in PAP without affecting LAP, systemic hemodynamics, renal function, or plasma AVP. Similarly, inflation of a balloon in the right atrium failed to alter renal function, plasma AVP, or PRA. Finally, constriction of the thoracic inferior vena cava decreased LAP and increased PRA. In summary, these data emphasize that inflation of a balloon in the left atrium of the conscious dog produces a composite response consisting of alterations in cardiovascular function, renal function, and circulating hormones. Moreover, our data indicate that the response is mediated by a reflex initiated from receptors located in the left atrium; we detected no evidence that receptors located in the pulmonary vasculature or right heart contribute to this response.


Dietary intake and major food sources of polyphenols in a Spanish population at high cardiovascular risk: the PREDIMED study.
Epidemiological data have shown an inverse association between the consumption of polyphenol-rich foods and the risk of cardiovascular disease or overall mortality. A comprehensive estimation of individual polyphenol intake in nutritional cohorts is needed to gain a better understanding of this association. The aim of this study was to estimate the quantitative intake of polyphenols and the major dietary sources in the PREDIMED (PREvención con DIeta MEDiterránea) cohort using individual food consumption records. The PREDIMED study is a large, parallel-group, multicentre, randomised, controlled 5-year feeding trial aimed at assessing the effects of the Mediterranean diet on the primary prevention of cardiovascular disease. A total of 7200 participants, aged 55-80 years, completed a validated 1-year food frequency questionnaire (FFQ) at baseline. Polyphenol consumption was calculated by matching food consumption data from the FFQ with the recently developed Phenol-Explorer database on polyphenol content in foods. The mean total polyphenol intake was 820 ± 323 mg day⁻¹ (443 ± 218 mg day⁻¹ of flavonoids and 304 ± 156 mg day⁻¹ of phenolic acids). Hydroxycinnamic acids were the phenolic group with the highest consumption and 5-caffeoylquinic acid was the most abundantly ingested individual polyphenol. The consumption of olives and olive oil was a differentiating factor in the phenolic profile of this Spanish population compared with other countries. In Mediterranean countries, such as Spain, the main dietary source of polyphenols is coffee and fruits, but the most important differentiating factor with respect to other countries is the consumption of polyphenols from olives and olive oil.