Inhibition of gonadotropin-stimulated ovarian steroid production by polyunsaturated fatty acids in teleost fish.
The effects of the polyunsaturated fatty acids (PUFAs)--eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and arachidonic acid (AA)--on in vitro steroid production by full-grown prematurational ovarian follicles from goldfish and rainbow trout were investigated. EPA and DHA inhibited gonadotropin-stimulated testosterone production in a dose-related manner, but AA was inhibitory only at the highest dose tested (400 microM). AA alone stimulated testosterone production by increasing cAMP production, but the effects of other PUFAs alone were marginal. The inhibitory actions by PUFAs were not restricted to long-chain PUFAs, as linoleic and linolenic acids had similar actions in the goldfish. The inhibitory action of EPA on testosterone production was reversible upon removal of the PUFA from medium. Testosterone production stimulated by the addition of the cAMP analogues, dibutyryl cAMP, and 8-bromo cAMP, was attenuated by PUFAs, suggesting that they act at a site distal to cAMP formation. A post-cAMP site regulating cholesterol availability may be involved as testosterone production induced by addition of 25OH-cholesterol was not affected by the PUFAs in either fish species. Together, these findings underscore the importance of lipids in ovarian physiology and suggest that PUFAs may participate in the regulation of ovarian steroidogenesis in teleost fish.


Chemotion-ELN part 2: adaption of an embedded Ketcher editor to advanced research applications.
The Ketcher editor, available as an Open Source software package for drawing chemical structures, has been expanded to include several features that allow storage, management and application of templates, as well as the use of symbols for a planning and processing of solid phase synthesis. In addition, tools for the drawing of coordinative bonds to represent e.g. organometallic compounds were added. The editor has been implemented into an Electronic Lab Notebook (ELN) application which enables the use of the Ketcher editor for advanced operations in chemistry research. The developments of the ELN-integrated Ketcher (ketcher-rails) support the retrieval of identifiers and structure-related information from external databases and the molecule-based calculation of analytical values. The reworked editor can be used to generate molecular structures in reaction templates and to generate syntheses plans.


Sabotage of antiviral signaling and effectors by influenza viruses.
Vertebrate cells activate multiple signaling modules upon virus infection to eliminate the invading pathogen and to prevent the establishment of a persistent infection. A major immediate response pathway is controlled by the RNA helicases RIG-I and MDA5, which, after recognition of viral nucleic acids, signal induction of the interferon (IFN)-alpha/beta cytokine family that upregulates numerous antiviral effector proteins. Virulent viruses, in contrast, have learned during co-evolution with their hosts to manipulate or avoid this response in order to prevail in a repulsive environment. Focusing on the influenza viruses and their IFN-antagonistic NS1 proteins, we summarize recent progress in this rapidly evolving field at the intersection of virology and immunobiology involving studies of how viral pathogens induce and sabotage cellular defenses.


Effect of Caffeine on Perceived Soreness and Functionality Following an Endurance Cycling Event.
Caldwell, AR, Tucker, MA, Butts, CL, McDermott, BP, Vingren, JL, Kunces, LJ, Lee, EC, Munoz, CX, Williamson, KH, Armstrong, LE, and Ganio, MS. Effect of caffeine on perceived soreness and functionality following an endurance cycling event. J Strength Cond Res 31(3): 638-643, 2017-Caffeine can reduce muscle pain during exercise; however, the efficacy of caffeine in improving muscle soreness and recovery from a demanding long-duration exercise bout has not been established. The purpose of this study was to investigate the effects of caffeine intake on ratings of perceived muscle soreness (RPMS) and perceived lower extremity functionality (LEF) following the completion of a 164-km endurance cycling event. Before and after cycling RPMS (1-to-6; 6 = severe soreness) and LEF (0-to-80; 80 = full functionality) were assessed by questionnaires. Subjects ingested 3 mg/kg body mass of caffeine or placebo pills in a randomized, double-blind fashion immediately after the ride and for the next 4 mornings (i.e., ∼800 hours) and 3 afternoons (i.e., ∼1200 hours). Before each ingestion, RPMS and LEF were assessed. Afternoon ratings of LEF were greater with caffeine ingestion the first day postride (65.0 ± 6.1 vs. 72.3 ± 6.7; for placebo and caffeine, respectively; p = 0.04), but at no other time points (p > 0.05). The caffeine group tended to have lower overall RPMS in the afternoon versus placebo (i.e., main effect of group; 1.1 ± 0.2 vs. 0.5 ± 0.2; p = 0.09). Afternoon RPMS for the legs was significantly lower in the caffeine group (main effect of caffeine; 1.3 ± 0.2 vs. 0.5 ± 0.3; p = 0.05). In conclusion, ingesting caffeine improved RPMS for the legs, but not LEF in the days following an endurance cycling event. Athletes may benefit from ingesting caffeine in the days following an arduous exercise bout to relieve feelings of soreness and reduced functionality.


Sleep Quality in Patients With Rotator Cuff Disease.
Little is known about the influence of rotator cuff pathology on sleep. The purpose of this study was to determine which patient-reported factors correlate with sleep disturbance in patients with rotator cuff disease. A nonrandomized, cross-sectional cohort study was performed to evaluate the effects of rotator cuff disease on sleep quality. Data collected at time zero (before any treatment) included the Single Assessment Numeric Evaluation rating, the American Shoulder and Elbow Surgeons score, the Pittsburgh Sleep Quality Index, patient demographics, and medical comorbidities. Statistical analysis included the Pearson correlation and multiple regression analysis to determine which patient-reported factors were associated with sleep disturbance. Nocturnal pain was reported by 91% of the 391 participants (274 with tendinitis and 117 with rotator cuff tears). Participants had a mean age of 57 years. Pearson correlation coefficients determined that poor sleep quality in one group or both the tendinitis and the rotator cuff tear groups was associated with higher pain visual analog scale scores (0.27 and 0.31; P = 0.004 and P < 0.0001, respectively), depression (0.27 and 0.30; P < 0.01), female sex (0.24 and 0.27; P < 0.001), presence of low back pain (0.25 and 0.27; P < 0.01), diabetes mellitus (0.24 in the rotator cuff tear group; P < 0.01), and increased body mass index (0.22 and 0.27; P = 0.02). The status of the rotator cuff did not correlate with increasing symptoms of shoulder pain or with worse sleep quality as measured by the Pittsburgh Sleep Quality Index. These results support the theory that worsening symptoms of shoulder pain may not be clearly associated with rotator cuff disease severity. Worse sleep quality scores in patients with rotator cuff disease are associated with pain, depression, female sex, low back pain, diabetes mellitus, and high body mass index. Overall, sleep quality did not differ among patients with varying rotator cuff disease severity. Only hypertension (in patients with rotator cuff tears) and concurrent cervical pathology (in patients with tendinitis) were uniquely related to the disease classification. Further investigation is needed to better define how these factors interact and influence nocturnal shoulder pain and sleep quality in patients with rotator cuff disease. Level III prognostic cohort study.


Characterizing the Quality of Insight by Interactions: A Case Study.
Understanding the quality of insight has become increasingly important with the trend of allowing users to post comments during visual exploration, yet approaches for qualifying insight are rare. This article presents a case study to investigate the possibility of characterizing the quality of insight via the interactions performed. To do this, we devised the interaction of a visualization tool-MediSyn-for insight generation. MediSyn supports five types of interactions: selecting, connecting, elaborating, exploring, and sharing. We evaluated MediSyn with 14 participants by allowing them to freely explore the data and generate insights. We then extracted seven interaction patterns from their interaction logs and correlated the patterns to four aspects of insight quality. The results show the possibility of qualifying insights via interactions. Among other findings, exploration actions can lead to unexpected insights; the drill-down pattern tends to increase the domain values of insights. A qualitative analysis shows that using domain knowledge to guide exploration can positively affect the domain value of derived insights. We discuss the study's implications, lessons learned, and future research opportunities.


Culture of Tumorigenic Cells on Protein Fibers Reveals Metastatic Cell Behaviors.
Tumorigenic cell behaviors can be suppressed or enhanced by their physicochemical environment. As a first step toward developing materials that allow tumorigenic behaviors to be observed and manipulated, we cultured related MCF10 breast cell lines on fibers composed of the Drosophila protein Ultrabithorax (Ubx). These cell lines, originally derived from fibrocystic breast tissue, represent a continuum of tumorigenic behavior. Immortal but nontumorigenic MCF10A cells, as well as semitumorigenic MCF10AT cells, attached and spread on Ubx fibers. MCF10CA-1a cells, the most highly transformed line, secreted high concentrations of matrix metalloproteinases when cultured on Ubx materials, resulting in differences in cell attachment and cytoskeletal structure, and enabling invasive behavior. Because the mechanical and functional properties of Ubx fibers can be genetically manipulated, these materials provide a valuable tool for cancer research, allowing creation of diverse microenvironments that allow assessment of invasive, metastatic behavior.


[Dynamic distribution of methandrostenolone in the body of white rats].
Dynamics of distribution of anabolic steroidal hormone methandrostenolone and routes of its elimination from the organism of Wistar rats were studied by using methods of radioisotopes and high-performance liquid chromatography. Methandrostenolone metabolites were shown to be excreted mainly in the urine. Methandrostenolone metabolism is a complicated process in the course of which redistribution of metabolites among various organs occurs. The anabolic effect of methandrostenolone is supposed to be due to the formation of its metabolites.


Depression symptoms are persistent in Type 2 diabetes: risk factors and outcomes of 5-year depression trajectories using latent class growth analysis.
To describe the long-term trajectories of depression symptom severity in people with Type 2 diabetes, and to identify predictors and associates of these trajectories. A community-dwelling cohort of 1201 individuals with Type 2 diabetes from the Fremantle Diabetes Study Phase II was followed for 5 years. The nine-item version of the Patient Health Questionnaire was administered annually to assess depression symptoms, and biomedical and psychosocial measures were assessed at baseline and biennially. Latent class growth analysis was used to identify classes of depression severity trajectories and associated outcomes, and logistic regression models were used to determine predictors of class membership. Three trajectories of depression symptoms were identified: continuously low depression symptoms (85.2%); gradually worsening symptoms that then began to improve (persistent depression - low-start; 7.3%); and gradually improving symptoms which later worsened (persistent depression - high-start; 7.5%). Younger age, being a woman, and a lifetime history of major depressive disorder, were associated with greater risk of persistent depression symptoms. Persistent depression was associated with consistently higher BMI over time, but not with changes in HbA1c or self-monitoring of blood glucose. A subset of individuals with Type 2 diabetes is at risk of depression symptoms that remain elevated over time. Younger, overweight individuals with a history of depression may benefit from early and intensive depression management and ongoing follow-up as part of routine Type 2 diabetes care.


Unlocking proteomic heterogeneity in complex diseases through visual analytics.
Despite years of preclinical development, biological interventions designed to treat complex diseases such as asthma often fail in phase III clinical trials. These failures suggest that current methods to analyze biomedical data might be missing critical aspects of biological complexity such as the assumption that cases and controls come from homogeneous distributions. Here we discuss why and how methods from the rapidly evolving field of visual analytics can help translational teams (consisting of biologists, clinicians, and bioinformaticians) to address the challenge of modeling and inferring heterogeneity in the proteomic and phenotypic profiles of patients with complex diseases. Because a primary goal of visual analytics is to amplify the cognitive capacities of humans for detecting patterns in complex data, we begin with an overview of the cognitive foundations for the field of visual analytics. Next, we organize the primary ways in which a specific form of visual analytics called networks has been used to model and infer biological mechanisms, which help to identify the properties of networks that are particularly useful for the discovery and analysis of proteomic heterogeneity in complex diseases. We describe one such approach called subject-protein networks, and demonstrate its application on two proteomic datasets. This demonstration provides insights to help translational teams overcome theoretical, practical, and pedagogical hurdles for the widespread use of subject-protein networks for analyzing molecular heterogeneities, with the translational goal of designing biomarker-based clinical trials, and accelerating the development of personalized approaches to medicine.


Reduced medium-term mortality following primary total hip and knee arthroplasty with an enhanced recovery program. A study of 4,500 consecutive procedures.
Our unit started to use routine multimodal techniques to enhance recovery for hip and knee arthroplasty in 2008. We have previously reported earlier discharge, a trend toward a reduction in complications, and a statistically significant reduction in mortality up to 90 days after surgery. In this study, we evaluated the same cohort to determine whether survival benefits were maintained at 2 years. We prospectively evaluated 4,500 unselected consecutive total hip and knee replacements. The first 3,000 underwent a traditional protocol (TRAD) and the later 1,500 underwent an enhanced recovery protocol (ER). Mortality data were collected from the Office of National Statistics (UK). There was a difference in death rate at 2 years (TRAD vs. ER: 3.8% vs. 2.7%; p = 0.05). Survival probability up to 3.7 years post surgery was significantly better in patients who underwent an ER protocol. This large prospective case series of unselected consecutive patients showed a reduction in mortality rate at 2 years following elective lower-limb hip and knee arthroplasty following the introduction of a multimodal enhanced recovery protocol. This survival benefit supports the routine use of an enhanced recovery program for hip and knee arthroplasty.


Serum levels and biochemical characteristics of cancer-associated antigen CA-549, a circulating breast cancer marker.
CA-549 is a circulating breast cancer-associated antigen that reacts with monoclonal antibody BC4E 549. Biochemical characterization of CA-549 revealed that it is an acidic (isoelectric point 5.2) glycoprotein that exhibits two bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions of apparent molecular weights of 400,000 and 512,000. Immunohistochemical staining of unfixed frozen tissue sections of human breast tumors and a variety of benign tissues with BC4E 549 revealed no preferential staining of tumor over benign breast tissue and cross-reactivity with a wide range of other benign tissues including kidney, liver, lung, colon, pancreas, ovary, and spleen. Serum levels of CA-549 were initially tested by an enzyme-linked immunosorbent assay inhibition using BC4E 549. This assay showed that CA-549 concentrations were elevated in 19 of 27 sera from patients with advanced breast cancer compared to 0 of 22 and 0 of 129 sera from benign breast disease patients and healthy females, respectively. These preliminary data suggested that CA-549 was a useful breast tumor marker; thus BC4E 549 was adapted to a sandwich immunoradiometric assay format suitable for routine use in the clinical laboratory and its performance was evaluated on a panel of 668 serum samples. The test detected significant concentrations of CA-549 in the sera of 40 of 80 patients with advanced breast cancer, 1 of 30 with early breast cancer, 4 of 19 with advanced lung cancer, 2 of 40 with advanced colon cancer, and 5 of 29 with advanced prostate cancer. The test showed a high degree of specificity, producing false-positives in only 3 of 79 benign breast patients, 2 of 25 benign liver patients, 2 of 70 benign colon patients, 2 of 19 benign lung patients, 0 of 20 benign prostate patients, and 3 of 257 healthy individuals. These data represent an overall 50% sensitivity and 98% specificity as a test for advanced breast cancer. These data indicate that this immunoradiometric assay is a useful test for the detection of circulating CA-549 in advanced breast cancer patients and suggest that it may prove useful as a monitor in the management of that disease.


Giving an account of one's pain in the anthropological interview.
In this paper, I analyze the illness stories narrated by a mother and her 13-year-old son as part of an ethnographic study of child chronic pain sufferers and their families. In examining some of the moral, relational and communicative challenges of giving an account of one's pain, I focus on what is left out of some accounts of illness and suffering and explore some possible reasons for these elisions. Drawing on recent work by Judith Butler (Giving an Account of Oneself, 2005), I investigate how the pragmatic context of interviews can introduce a form of symbolic violence to narrative accounts. Specifically, I use the term "genre of complaint" to highlight how anthropological research interviews in biomedical settings invoke certain typified forms of suffering that call for the rectification of perceived injustices. Interview narratives articulated in the genre of complaint privilege specific types of pain and suffering and cast others into the background. Giving an account of one's pain is thus a strategic and selective process, creating interruptions and silences as much as moments of clarity. Therefore, I argue that medical anthropologists ought to attend more closely to the institutional structures and relations that shape the production of illness narratives in interview encounters.


Analysis of the effect of formulation properties and process parameters on granule formation in twin-screw wet granulation.
In the last few years, twin-screw wet granulation (TSWG) has become one of the key continuous pharmaceutical unit operations. Despite the many studies that have been performed, only little is known about the effect of the starting material properties on the stepwise granule formation along the length of the twin-screw granulator (TSG) barrel. Hence, this study obtained a detailed understanding of the effect of formulation properties (i.e., Active Pharmaceutical Ingredient (API) properties, formulation blend particle size distribution and formulation drug load) and process settings on granule formation in TSWG. An experimental set-up was used allowing the collection of granules at the different TSG compartments. Granules were characterized in terms of granule size, shape, binder liquid and API distributions. Liquid-to-solid (L/S) ratio was the only TSG process parameter impacting the granule size and shape evolution. Particle size and flow properties (e.g., flow rate index) had an important effect on the granule size and shape changes whereas water-related properties (e.g., water binding capacity and solubility) became influential at the last TSG compartments. The API solubility and L/S ratio were found to have a major impact on the distribution of binder liquid over the different granule size fractions. In the first TSG compartment (i.e., wetting compartment), the distribution of the API in the granules was influenced by its solubility in the granulation liquid.


Dual-axis confocal microscope for high-resolution in vivo imaging.
We describe a novel confocal microscope that uses separate low-numerical-aperture objectives with the illumination and collection axes crossed at angle theta from the midline. This architecture collects images in scattering media with high transverse and axial resolution, long working distance, large field of view, and reduced noise from scattered light. We measured transverse and axial (FWHM) resolution of 1.3 and 2.1 microm, respectively, in free space, and confirm subcellular resolution in excised esophageal mucosa. The optics may be scaled to millimeter dimensions and fiber coupled for collection of high-resolution images in vivo.


Discovering and Applying the Urban Rules of Life to Design Sustainable and Healthy Cities.
The city and its urban biome provides an extreme laboratory for studying fundamental biological questions and developing best practices for sustaining biodiverse and well-functioning ecological communities within anthropogenic built environments. We propose by studying urban organisms, urban biotic communities, the urban biome, and the interactions between the urban biome and peri-urban built and natural environments, we can (1) discover new "rules of life" for the structure, function, interaction, and evolution of organisms; (2) use these discoveries to understand how novel emerging biotic communities affect and are affected by anthropogenic environmental changes in climate and other environmental factors; and (3) apply what we have learned to engage residents of the urban biome, and design cities that are more biologically diverse, are provided with more and better ecosystem services, and are more equitable and healthier places to live. The built environment of the urban biome is a place that reflects history, economics, technology, governance, culture, and values of the human residents; research on and applications of the rules of life in the urban biome can be used by all residents in making choices about the design of the cities where they live. Because inhabitants are directly invested in the environmental quality of their neighborhoods, research conducted in and about the urban environment provides a great opportunity to engage wide and diverse communities of people. Given the opportunity to engage a broad constituency-from basic researchers to teachers, civil engineers, landscape planners, and concerned citizens-studying the translation of the rules of life onto the urban environment will result in an integrative and cross-cutting set of questions and hypotheses, and will foster a dialog among citizens about the focus of urban biome research and its application toward making more equitable, healthy, livable, sustainable, and biodiverse cities.


gcd12 mutations are gcn3-dependent alleles of GCD2, a negative regulator of GCN4 in the general amino acid control of Saccharomyces cerevisiae.
GCD12 encodes a translational repressor of the GCN4 protein, a transcriptional activator of amino acid biosynthetic genes in the yeast Saccharomyces cerevisiae. gcd12 mutations override the requirement for the GCN2 and GCN3 gene products for derepression of GCN4 expression, suggesting that GCN2 and GCN3 function indirectly as positive regulators by negative regulation of GCD12. In addition to their regulatory phenotype, gcd12 mutants are temperature-sensitive for growth (Tsm-) and, as shown here, deletion of the GCD12 gene is unconditionally lethal. Both the regulatory and the Tsm- phenotypes associated with gcd12 point mutations are completely overcome by wild-type GCN3, implying that GCN3 can promote or partially substitute for the functions of GCD12 in normal growth conditions even though it antagonizes GCD12 regulatory function in starvation conditions. The GCD12 gene has been cloned and mapped to the right arm of chromosome VII, very close to the map position reported for GCD2. We demonstrate that GCD12 and GCD2 are the same genes; however, unlike gcd12 mutations, the growth defect and constitutive derepression phenotypes associated with the gcd2-1 mutation are expressed in the presence of the wild-type GCN3 gene. These findings can be explained by either of two alternative hypotheses: (1) gcd12 mutations affect a domain of the GCD2 protein that directly interacts with GCN3, and complex formation stabilizes mutant gcd12 (but not gcd2-1) gene products; (2) gcd12 mutations selectively impair one function of GCD2 that is replaceable by GCN3, whereas gcd2-1 inactivates a different GCD2 function for which GCN3 cannot substitute. Both models imply a close interaction between these two positive and negative regulators in general amino acid control.


Effects of long-acting testosterone undecanoate on bone mineral density in middle-aged men with late-onset hypogonadism and metabolic syndrome: results from a 36 months controlled study.
We evaluated the effects of long-term testosterone replacement therapy (TRT) on the bone mineral density (BMD) in obese patients with metabolic syndrome (MS) and late-onset hypogonadism (LOH). Sixty men (mean age 57 ± 10) with low serum testosterone (T < 320 ng/dL) and MS regardless the presence of osteoporosis were enrolled. Forty men received intramuscular T-undecanoate (TU) four times/year for 36 months and 20 age-matched hypogonadal men with MS in whom T treatment was contraindicated were used as controls. Hormonal, biochemical markers, vertebral and femoral BMD by dual-energy x-ray absorptiometry were measured. At baseline, overall patients had mild osteopenia (lumbar BMD= 0.891 ± 0.097 g/cm(2); femoral BMD= 0.847 ± 0.117 g/cm(2)). TU induced a significant improvement of bone mass after 36 months (lumbar BMD=1.053 ± 0.145 g/cm(2); p < 0.002; femoral BMD=0.989 ± 0.109; p < 0.003 g/cm(2)) with a 5%/year increase and a significant reduction in hs-CRP without changes in body mass index. A direct relationship between serum T and BMD increments at the lumbar (r(2) = 0.66, p < 0.0001) and femoral (r(2) =0.52, p < 0.0001) sites was demonstrated. Study adherence was 50% without serious side effects. Long-term TRT in middle-aged men with LOH and MS determines a significant increase in both vertebral and femoral BMD related to increased serum T levels, probably independently from estradiol modifications.


The Endoscopic Removal of Eroded Foreign Bodies in the Ureter.
A complication of using foreign materials in surgery is potential erosion into nearby tissues. The endoscopic removal of foreign bodies that have eroded into the urinary tract is a safe and minimally invasive option that has previously been described, most commonly in the bladder and urethra. We present the case of a patient who had a remote history of a pyeloplasty and was found to have different foreign bodies eroding into the ureter causing symptoms. To our knowledge, this is the first case where a patient presented with two different types of ureteral foreign body erosions that were each effectively treated endoscopically.


Perceived risk from occupational stress: a survey of 15 European countries.
To examine the ranking of workers in 15 European countries with respect to the extent that work is perceived to influence occupational stress. International interviewer administered cross-sectional survey. Control variables included demographics, perceived job conditions, and job satisfaction. A sample of working adults (n = 11 054) from 15 European countries were studied. In three statistical models, the British sample ranked in the lower half of the distribution, where higher rankings indicated greater perceived risk of occupational stress. The Greek sample was ranked first in all models, and the Irish sample ranked last in all models. Since there is sociocultural variation in the perceived risk from occupational stress, it is possible that sociocultural factors influence the accuracy of national monitoring systems and the success of occupational health policy in this area.


Exercise and insulin cause GLUT-4 translocation in human skeletal muscle.
Studies in rodents have established that GLUT-4 translocation is the major mechanism by which insulin and exercise increase glucose uptake in skeletal muscle. In contrast, much less is known about the translocation phenomenon in human skeletal muscle. In the current study, nine healthy volunteers were studied on two different days. On one day, biopsies of vastus lateralis muscle were taken before and after a 2-h euglycemic-hyperinsulinemic clamp (0.8 mU. kg(-1). min(-1)). On another day, subjects exercised for 60 min at 70% of maximal oxygen consumption (VO(2 max)), a biopsy was obtained, and the same clamp and biopsy procedure was performed as that during the previous experiment. Compared with insulin treatment alone, glucose infusion rates were significantly increased during the postexercise clamp for the periods 0-30 min, 30-60 min, and 60-90 min, but not during the last 30 min of the clamp. Plasma membrane GLUT-4 content was significantly increased in response to physiological hyperinsulinemia (32% above rest), exercise (35%), and the combination of exercise plus insulin (44%). Phosphorylation of Akt, a putative signaling intermediary for GLUT-4 translocation, was increased in response to insulin (640% above rest), exercise (280%), and exercise plus insulin (1,000%). These data demonstrate that two normal physiological conditions, moderate intensity exercise and physiological hyperinsulinemia approximately 56 microU/ml, cause GLUT-4 translocation and Akt phosphorylation in human skeletal muscle.


A Model for the Spatiotemporal Design of Gene Regulatory Circuits †.
Mathematical modeling assists the design of synthetic regulatory networks by providing a detailed mechanistic understanding of biological systems. Models that can predict the performance of a design are fundamental for synthetic biology since they minimize iterations along the design-build-test lifecycle. Such predictability depends crucially on what assumptions (i.e., biological simplifications) the model considers. Here, we challenge a common assumption when it comes to the modeling of bacterial-based gene regulation: considering negligible the effects of intracellular physical space. It is commonly assumed that molecules, such as transcription factors (TF), are homogeneously distributed inside a cell, so there is no need to model their diffusion. We describe a mathematical model that accounts for molecular diffusion and show how simulations of network performance are decisively affected by the distance between its components. Specifically, the model focuses on the search by a TF for its target promoter. The combination of local searches, via one-dimensional sliding along the chromosome, and global searches, via three-dimensional diffusion through the cytoplasm, determine TF-promoter interplay. Previous experimental results with engineered bacteria in which the distance between TF source and target was minimized or enlarged were successfully reproduced by the spatially resolved model we introduce here. This suggests that the spatial specification of the circuit alone can be exploited as a design parameter in synthetic biology to select programmable output levels.


Synthesis for an electrooptic light scanner with an arbitrary beam shape.
Two methods are described for synthesizing the electrooptic light scanner with an arbitrarily prescribed one-dimensional spatial distribution of the light beam. The electrooptic light scanner is constructed by a series of cascaded stages, each stage containing a natural-birefringent electrooptic crystal with an inclination angle between opposite faces. One method is realized by a set of cascaded stages, each of which contains a polarizer, electrooptic crystal, optical compensator, and analyzer; and the other method by a multistage construction that is inserted between the input and output polarizers, each stage being composed of only an electrooptic crystal and optical compensator. Two examples for each method are given to illustrate the synthesis procedures.


Impact of isomeric structures on transistor performances in naphthodithiophene semiconducting polymers.
Four isomeric naphthodithiophenes (NDTs) with linear and angular shapes were introduced into the polythiophene semiconductor backbones, and their field-effect transistor performances were characterized. The polymers bearing naphtho[1,2-b:5,6-b']dithiophene (NDT3), an angular-shaped NDT, exhibited the highest mobilities of ∼0.8 cm(2) V(-1) s(-1) among the four NDT-based polymers, which is among the highest reported so far for semiconducting polymers. Interestingly, the trend of the mobility in the NDT-based polymers was contrary to our expectations; the polymers with angular NDTs showed higher mobilities than those with linear NDTs despite the fact that naphtho[2,3-b:6,7-b']dithiophene (NDT1), a linear-shaped NDT, has shown the highest mobility in small-molecule systems. X-ray diffraction studies revealed that angular-NDT-based polymers gave the highly ordered structures with a very close π-stacking distance of 3.6 Å, whereas linear-NDT-based polymers had a very weak or no π-stacking order, which is quite consistent with the trend of the mobility. The nature of such ordering structures can be well understood by considering their molecular shapes. In fact, a linear NDT (NDT1) provides angular backbones and an angular NDT (NDT3) provides a pseudostraight backbone, the latter of which can pack into the highly ordered structure and thus facilitate the charge carrier transport. In addition to the ordering structure, the electronic structures seem to correlate with the carrier transport property. MO calculations, supported by the measurement of ionization potentials, suggested that, while the HOMOs are relatively localized within the NDT cores in the linear-NDT-based polymers, those are apparently delocalized along the backbone in the angular-NDT-based polymers. The latter should promote the efficient HOMO overlaps between the polymer backbones that are the main paths of the charge carrier transport, which also agrees with the trend of the mobility. With these results, we conclude that angular NDTs, in particular NDT3, are promising cores for high-performance semiconducting polymers. We thus propose that both the molecular shapes and the electronic structures are important factors to be considered when designing high performance semiconducting polymers.


Gamma knife radiosurgery in children.
52 pediatric patients were treated with radiosurgery at the University of California, San Francisco. Arteriovenous malformations were treated in 27 patients. Complete obliteration was noted in 4 of 12 patients imaged more than 2 years after radiosurgery. Arteriovenous malformation rebleed was noted in 1 patient. Symptomatic T2 changes were noted in 2 patients. Among 29 neoplasms treated in 25 patients, local control was noted in 5 of 7 low-grade gliomas, 5 of 14 high-grade gliomas, 4 of 5 craniopharyngiomas and 3 of 3 sarcomas. Three patients treated for neoplasms developed necrosis after radiosurgery.


Sexual selection in seaweed? Testing Bateman's principles in the red alga Gracilaria gracilis.
In anisogamous species, sexual selection is expected to be stronger in males. Bateman's principles state that the variance in (i) reproductive and (ii) mating success is greater for males, and (iii) the relationship between reproductive success and mating success (the Bateman gradient) is also stronger for males than for females. Sexual selection, based on Bateman's principles, has been demonstrated in animals and some angiosperms, but never in a seaweed. Here we focus on the oogamous haploid-diploid rhodophyte Gracilaria gracilis in which previous studies have shown evidence for non-random mating, suggesting the existence of male-male competition and female choice. We estimated mating and reproductive success using paternity analyses in a natural population where up to 92% of fertilizations occurred between partners of that population. The results show that the variance in mating success is significantly greater in males than in females and that the Bateman gradient is positive only in males. Distance to female partners also explains a minor part of the variance in male mating success. Although there is no evidence for sexual dimorphism, our study supports the hypothesis that sexual selection occurs in G. gracilis, probably on male traits, even if we cannot observe, characterize or quantify them yet.


Adherence to advance directives in critical care decision making: vignette study.
To explore health professionals' decision making in a critical care scenario when there is an advance directive. Qualitative study. Scotland. Interviewees (n = 12) comprising general practitioners, hospital specialists, and nurses, and six focus groups (n = 34 participants) comprising general practitioners, geriatricians (consultants and specialist registrars), hospital nurses, and hospice nurses. When presented with an advance directive that applied to the same hypothetical scenario, health professionals came to divergent conclusions as to the "right thing to do." Arguments opposing treatment centred on the supremacy of autonomy as an ethical principle. Other arguments were that the decision to treat was consistent with the terms of the advance directive, or that, notwithstanding the advance directive, the patient's quality of life was sufficient to warrant treatment. Advance directives are open to widely varying interpretation. Some of this variability is related to the ambiguity of the directive's terminology whereas some is related to the willingness of health professionals to make subjective value judgments concerning quality of life.


Transcapho perilunate dislocation with palmar extrusion of the scaphoid proximal pole.
Perilunate fracture-dislocations usually combine ligament ruptures, bone avulsions, and fractures in different patterns. Rarely a displaced fracture of the scaphoid can coexist with a scapho-lunate dissociation and can result in enucleation of the proximal pole. We report about a case of trans-scaphoid perilunate dislocation with palmar extrusion/enucleation of the scaphoid proximal pole, treated with scaphoid fracture open reduction and internal fixation with screw, scapho-lunate ligament repair with an anchor and vascularization of the scaphoid proximal pole with the 2nd intermetacarpal artery. At 52 months follow up we had good clinical and radiographic results. In conclusion, scientific literature including our experience about this rare complex lesion of the wrist is too weak to support an effective strategy of management but we think that the careful analysis of the single problems can be the key to solve the complexity. Goal of the treatment should be complete revascularization and healing of the scaphoid, avoiding non union and avascular necrosis; simultaneously a proper ligament reconstruction is fundamental to re-establish carpal stability. Prevention of carpal collapse for a SNAC o SLAC situation is essential to reach a good level of Quality of Life and satisfaction of the patient.


A compact, versatile inhalation exposure chamber for small animal studies.
An inhalation exposure chamber was designed to fit conveniently into a standard laboratory exhaust hood. The chamber was constructed entirely of non-corrosive materials and was suitable for continuous exposure of 6 small laboratory animals to corrosive atmospheres such as ozone. A separate, outer chamber surrounding a smaller, inner chamber permitted access to each individually housed animal without disturbing the experimental atmosphere. Facilities were available for ad libitum access to food and water for chronic exposures. The chamber was easily disassembled for cleaning. A safety mechanism was described which automatically stops the generation of a toxic gas in the event of an exhaust system failure.


Requirement of varicella-zoster virus immediate-early 4 protein for viral replication.
Varicella-zoster virus (VZV) is an alphaherpesvirus that causes two diseases, chickenpox and zoster. VZV open reading frame 4 (ORF4) encodes the immediate-early 4 (IE4) protein, which is conserved among alphaherpesvirus and has transactivation activity in transient transfections. To determine whether the ORF4 gene product is essential for viral replication, we used VZV cosmids to remove ORF4 from the VZV genome. Deleting ORF4 was incompatible with recovery of infectious virus, whereas transfections done by using repaired cosmids with ORF4 inserted at a nonnative site yielded virus. To analyze the functional domain of IE4, we introduced a mutation altering the C-terminal amino acids, KYFKC (K443S), which was designed to disrupt the dimerization of IE4 protein. Transfections with these mutant cosmids yielded no virus, indicating that this KYFKC motif was essential for IE4 function.


[Use of modified intrauterine device (IUD) TCu 380 with chromium filaments in the immediate postpartum].
Postpartum insertion of the intrauterine device (IUD) can provide an effective and convenient means of contraception. As a result, the use of IUD's has steadily increased, and the Family Planning Program recommends it because it offers many advantages. However, a major risk associated with the use of IUD's is the possibility of its expulsion, which ranges from 4 to 60%. Furthermore, 20% of women who expelled IUD's were unaware that it occurred, thus increasing their susceptibility to unwanted pregnancies. IUD's modified by the addition of biodegradable strands of chromic suture at time of implantation are thought to be less likely to be expelled, as the sutures anchor the IUD more firmly to the endometrium, and it is possible that the use of these sutures may decrease the risk of expulsion. Therefore we determined if the chromic extension to IUD enhance retention and decreased the rate of expulsion. In one year of study, 150 women received a modified IUD (TCu 380) with chromic catgut number 0 (ccO) in the transversal arm within 10 minutes of delivery of the placenta. However, only 84 women completed the follow-up study. To evaluate IUD expulsion, exploratory examinations were conducted during the immediate postpartum, at 7 days postpartum and at 6 weeks after delivery. We determined the presence or expulsion of the IUD in these three periods postpartum. Other parameters such as parity, age and marital status were also considered. A total of 14 modified IUD's (16.6%) were expelled. This represents a similar frequency of expulsion both during the immediate postpartum and the 7 days postpartum period. There was no significant difference in the rate of expulsion between the two periods. Moreover, there was no expulsion at 6 weeks postpartum. The primiparity women had the highest percentage of expulsion (22.8%). Single mothers either living with or without their sexual partner had expulsion rates of 20.6 and 20.5%, respectively. The highest rate of expulsion when we considered age was in women younger than 30 years of age. There was only one case of expulsion where the mother was unaware of its occurrence. The results described here indicate that age, parity and civil status have no direct influence on expulsion of modified IUD's with chrome extensions. In all postpartum periods studied the p-value was greater than 0.05. Six weeks postpartum was the only time at which there was no IUD expulsion. This is most likely a consequence of decreased uterine cavity size and closure cervix. The modified IUD TCu 380 with chrome strands shows a rate of expulsion similar to other modified IUD's as evidenced in the literature. However, there is a lower rate of expulsion than in unmodified IUD's. It is an important to rote that incomplete expulsion of modified IUD's in this study was considered a total expulsion which represent loss of contraceptive effectiveness. On the contrary, if we had considered only completely expelled IUD's in this study, the percentage would have been significantly lower, as only one total expulsion was found. Further studies are necessary to investigate consecutive expulsions of the TCu 380 modified IUD.


Diabetic Retinopathy: The Role of Mitochondria in the Neural Retina and Microvascular Disease.
Diabetic retinopathy (DR), a common chronic complication of diabetes mellitus and the leading cause of vision loss in the working-age population, is clinically defined as a microvascular disease that involves damage of the retinal capillaries with secondary visual impairment. While its clinical diagnosis is based on vascular pathology, DR is associated with early abnormalities in the electroretinogram, indicating alterations of the neural retina and impaired visual signaling. The pathogenesis of DR is complex and likely involves the simultaneous dysregulation of multiple metabolic and signaling pathways through the retinal neurovascular unit. There is evidence that microvascular disease in DR is caused in part by altered energetic metabolism in the neural retina and specifically from signals originating in the photoreceptors. In this review, we discuss the main pathogenic mechanisms that link alterations in neural retina bioenergetics with vascular regression in DR. We focus specifically on the recent developments related to alterations in mitochondrial metabolism including energetic substrate selection, mitochondrial function, oxidation-reduction (redox) imbalance, and oxidative stress, and critically discuss the mechanisms of these changes and their consequences on retinal function. We also acknowledge implications for emerging therapeutic approaches and future research directions to find novel mitochondria-targeted therapeutic strategies to correct bioenergetics in diabetes. We conclude that retinal bioenergetics is affected in the early stages of diabetes with consequences beyond changes in ATP content, and that maintaining mitochondrial integrity may alleviate retinal disease.


[Propagation of the pathogen of Lyme disease Borrelia burgdorferi in the ticks Ixodes persulcatus].
An increase of the number of Borrelia burgdorferi s. l. in nymphs was observed 3-4 months later the moulting and during the following 4-5 months. The percent of infected ticks remained unchanged. The reproduction of spirochete was going not only at a room temperature, but also at a lower temperature, 3-5 degrees (imitation of hibernation conditions). An increase of the spirochete density was especially well marked, when the ticks were maintained under fluctuated daily conditions. We have found out the difference in the transphase transmission. Almost all larvae and nymphs infected by a bloodsucking retained spirochetes after moulting (the first transphase transmission). When the nymphs infected at larval stage were fed on an uninfected mouse, only 30% of imago ticks retained the spirochetes (the second transphase transmission).


Multiple Augmented Reduced Rank Regression for Pan-Cancer Analysis.
Statistical approaches that successfully combine multiple datasets are more powerful, efficient, and scientifically informative than separate analyses. To address variation architectures correctly and comprehensively for high-dimensional data across multiple sample sets (i.e., cohorts), we propose multiple augmented reduced rank regression (maRRR), a flexible matrix regression and factorization method to concurrently learn both covariate-driven and auxiliary structured variation. We consider a structured nuclear norm objective that is motivated by random matrix theory, in which the regression or factorization terms may be shared or specific to any number of cohorts. Our framework subsumes several existing methods, such as reduced rank regression and unsupervised multi-matrix factorization approaches, and includes a promising novel approach to regression and factorization of a single dataset (aRRR) as a special case. Simulations demonstrate substantial gains in power from combining multiple datasets, and from parsimoniously accounting for all structured variation. We apply maRRR to gene expression data from multiple cancer types (i.e., pan-cancer) from TCGA, with somatic mutations as covariates. The method performs well with respect to prediction and imputation of held-out data, and provides new insights into mutation-driven and auxiliary variation that is shared or specific to certain cancer types.


Dicer1 activity in the stromal compartment regulates nephron differentiation and vascular patterning during mammalian kidney organogenesis.
MicroRNAs, activated by the enzyme Dicer1, control post-transcriptional gene expression. Dicer1 has important roles in the epithelium during nephrogenesis, but its function in stromal cells during kidney development is unknown. To study this, we inactivated Dicer1 in renal stromal cells. This resulted in hypoplastic kidneys, abnormal differentiation of the nephron tubule and vasculature, and perinatal mortality. In mutant kidneys, genes involved in stromal cell migration and activation were suppressed as were those involved in epithelial and endothelial differentiation and maturation. Consistently, polarity of the proximal tubule was incorrect, distal tubule differentiation was diminished, and elongation of Henle's loop attenuated resulting in lack of inner medulla and papilla in stroma-specific Dicer1 mutants. Glomerular maturation and capillary loop formation were abnormal, whereas peritubular capillaries, with enhanced branching and increased diameter, formed later. In Dicer1-null renal stromal cells, expression of factors associated with migration, proliferation, and morphogenic functions including α-smooth muscle actin, integrin-α8, -β1, and the WNT pathway transcriptional regulator LEF1 were reduced. Dicer1 mutation in stroma led to loss of expression of distinct microRNAs. Of these, miR-214, -199a-5p, and -199a-3p regulate stromal cell functions ex vivo, including WNT pathway activation, migration, and proliferation. Thus, Dicer1 activity in the renal stromal compartment regulates critical stromal cell functions that, in turn, regulate differentiation of the nephron and vasculature during nephrogenesis.


[Estimated equivalent radiation conditions (EERC), more useful clinical data and estimation of potential resorption of lesion nodule with respect to its volume and total focal dose].
Assumptions on properties of tumor cells are defined. On the basis on them the survival rate of tumor cells is described by the LQ function and the dependence of resorption probability (RP) in the lesion nodule on the number survived tumor cells (tumor volume) and on the total focal dose (TFD) is described by the Poisson function. An analysis of the above approach to the determination of lesion-nodule RP resulted in designing a calculation method for EERC that can be used to estimate the lesion-nodule RP as a function of its volume and TFD. An analytical or graphical description of the RP/TFD dependence for a fixed volume of lesion nodule is needed for the implementation of EERC. There is no need to determine the cell structure of tissue or the radiobiological properties of its cells. The possibility to construct the RP section function and to extend, thus, the volume of useful clinical data belongs to the method's advantages.


Perceived vaccination status in ecotourists and risks of anthropozoonoses.
Anthropozoonotic (human to nonhuman animal) transmission of infectious disease poses a significant threat to wildlife. A large proportion of travelers to tropical regions are not protected against vaccine-preventable illnesses, and a majority of these travelers demonstrate poor recall of actual vaccination status. Here we characterize self-perceived vaccination status among a large sample of ecotourists at the Sepilok Orangutan Rehabilitation Centre, Sabah, Malaysia. Despite their recognized travel itinerary to view endangered animals, tourists at wildlife sanctuaries are not adequately protected against vaccine-preventable illnesses. Of 633 surveys, over half reported being currently vaccinated against tuberculosis, hepatitis A, hepatitis B, polio, and measles. Fewer participants reported current vaccination status for influenza, rabies, and chickenpox. Despite the fact that the majority of visitors to Sepilok are from temperate regions where influenza is relatively more prevalent, 67.1% of those surveyed with medical-related occupations reported not being currently vaccinated for influenza. Ecotourists concerned about environmental protection are themselves largely unaware of their potential contribution to the spread of diseases to animals. The risks of negatively affecting animal populations must be communicated to all concerned parties, and this may begin by urging travelers to examine their actual vaccination status, particularly as the ecotourism industry continues its rapid expansion, and is seen increasingly as a possible tool to save great ape populations from extinction.


Glutamine Modulates Macrophage Lipotoxicity.
Obesity and diabetes are associated with excessive inflammation and impaired wound healing. Increasing evidence suggests that macrophage dysfunction is responsible for these inflammatory defects. In the setting of excess nutrients, particularly dietary saturated fatty acids (SFAs), activated macrophages develop lysosome dysfunction, which triggers activation of the NLRP3 inflammasome and cell death. The molecular pathways that connect lipid stress to lysosome pathology are not well understood, but may represent a viable target for therapy. Glutamine uptake is increased in activated macrophages leading us to hypothesize that in the context of excess lipids glutamine metabolism could overwhelm the mitochondria and promote the accumulation of toxic metabolites. To investigate this question we assessed macrophage lipotoxicity in the absence of glutamine using LPS-activated peritoneal macrophages exposed to the SFA palmitate. We found that glutamine deficiency reduced lipid induced lysosome dysfunction, inflammasome activation, and cell death. Under glutamine deficient conditions mTOR activation was decreased and autophagy was enhanced; however, autophagy was dispensable for the rescue phenotype. Rather, glutamine deficiency prevented the suppressive effect of the SFA palmitate on mitochondrial respiration and this phenotype was associated with protection from macrophage cell death. Together, these findings reveal that crosstalk between activation-induced metabolic reprogramming and the nutrient microenvironment can dramatically alter macrophage responses to inflammatory stimuli.


EUS-FNA and FDG-PET are complementary procedures in the diagnosis of enlarged mediastinal lymph nodes.
Transoesophageal endosonography with fine needle aspiration (EUS-FNA) and 2-deoxy-2-[fluorine-18]fluoro-D-glucose positron emission tomography (FDG-PET) are now standard diagnostic procedures of the mediastinum. Our aim was to compare their value in the assessment of enlarged mediastinal lymph nodes detected by computed tomography. Forty consecutive patients with a suspicion of cancer or a history of pulmonary, digestive, urogenital or mammary neoplasia and presenting with supracentimetric lymph nodes on computed tomography underwent whole body FDG-PET and EUS-FNA. Final diagnosis of malignancy was obtained by cytology, surgery or long-term follow-up. EUS-FNA showed a sensitivity, specificity and accuracy for detection of malignancy of 793, 100 and 85%, respectively. The biopsy material was adequate for cytological examination in 37 patients. Sensitivity, specificity and accuracy of PET were 100, 54.5 and 87.5%, respectively. FDG-PET correctly diagnosed the primary site in 27 patients, and showed additional unknown extrathoracic metastatic sites in 15 patients. The five false positive results observed with FDG-PET consisted in a final diagnosis of sarcoidosis, tuberculosis, anthracosilicosis and reactive lymph nodes, respectively. The association of FDG-PET and EUS-FNA avoided more invasive procedures (mediastinoscopies or staging surgery) in 34 patients. EUS-FNA and FDG-PET are complementary diagnostic procedures combining the high sensitivity of FDG-PET and the high specificity of EUS-FNA to accurately diagnose malignancy in enlarged mediastinal lymph nodes identified by CTscan. The combination of the two procedures in selected cases with pulmonary cancer or extra-thoracic tumours avoided more invasive diagnostic and surgical procedures.


Application of quinone-based fluorophore and native fluorescence for the spectrofluorimetric determination of agomelatine in dosage form: Identification of acidic and alkaline- induced degradation products by LC-MS/TOF.
Three spectrofluorimetric methods were developed for agomelatine (AGM) determination in commercial tablets. Method A is based on measuring the native fluorescence of AGM aqueous solution at 230/360 nm. Methods B and C are based on the formation of a charge transfer complex between AGM and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and 7,7,8,8-tetracyanoquinodimethane (TCNQ) with measurement of the formed fluorophore at 365/475 nm and 250/304 nm, respectively. The relative fluorescence intensity (RFI) of AGM-DDQ complex was greatly enhanced in the presence of methyl-β-cyclodextrin (CD). The methods were linear over the concentration ranges of 0.015-0.5, 0.5-8.0, 0.09-6.0 and 0.05-0.2 μg/ml for AGM-native fluorescence, AGM-DDQ, AGM-DDQ-CD and AGM-TCNQ complexes, respectively with excellent correlation coefficients (r = 0.9999). The methods were validated as per the International Conference on Harmonization (ICH) guidelines and all validation requirements were satisfied. The developed methods were extended to the analysis of AGM in commercial tablets. Furthermore, the stability of AGM was studied under different stress conditions (alkaline, acidic, oxidative and photolytic). The potential alkaline and acidic degradation products were identified by LC-MS/TOF.


Epidemiology and treatment of Bell's palsy in children in northern Taiwan.
Bell's palsy is not uncommon in children. This study was performed to evaluate the epidemiology of Bell's palsy in the northern Taiwanese pediatric population, and the effectiveness of corticosteroid treatment. The medical records of pediatric patients with a primary diagnosis of facial palsy from April 2002 through March 2007 were reviewed. Patients with secondary facial palsy were excluded from the analysis. 289 episodes of facial palsy were identified and the clinical findings of 134 episodes among 132 patients were assessed. The median +/- standard deviation age was 9.9 +/- 4.9 years, and 58.2% of patients were girls. Children were more likely to have episodes of Bell's palsy during the cold season, with a peak in January. The left (67 episodes; 50.0%) and right (64 episodes; 47.8%) facial nerves were involved with similar frequency. Common symptoms were postauricular pain (11.2%) and facial hypoesthesia (9.0%). Of 51 episodes of Bell's palsy with complete follow-up, corticosteroids were given for 44 episodes. Thirty eight patients (86.4%) given corticosteroids had complete recovery and 4 patients (57.1%) recovered without corticosteroids. Rates of complete recovery did not differ significantly between the 2 groups (p = 0.08). There were no significant differences in the recovery rate between early (< or = 3 days) and late (4-7 days) administration. In northern Taiwan, childhood Bell's palsy peaks from January through March. The majority of children with Bell's palsy recovered completely. There was no significant effect of corticosteroid treatment for children with Bell's palsy.


Familial lung disease associated with proliferation and desquamation of type II pneumonocytes.
A family is described in which certain infants presented with a diffuse parenchymal pulmonary disorder characterized by proliferation and desquamation of alveolar epithelial cells that progressed to severe pulmonary fibrosis and cystic dysplasia, resulting in "honeycomb" lung typical of chronic pulmonary interstitial disease (idiopathic pulmonary fibrosis) at death. Electron microscopic examination of a lung biopsy specimen from one infant revealed that the proliferating/desquamating cells were ultrastructurally normal-appearing type II pneumonocytes with osmiophilic lamellar bodies. This patient did not respond to prednisone therapy, but she showed great improvement with chloroquine therapy, which was confirmed by a repeated lung biopsy. Possible mechanisms that might account for the effect of chloroquine, such as inhibition of DNA synthesis, as well as the cellular pathobiology of type II pneumonocyte proliferation in this disease, are discussed.


Multiple myeloma with concurrent immune thrombocytopenic purpura.
Multiple myeloma (MM) is the second most common haematological malignancy in the USA. MM has been linked to various autoimmune disorders in many studies; one systemic review even suggested an increased risk of MM among patients with autoimmune disorders. MM is associated with many haematological, rheumatologic and neurological conditions. A few case reports suggest that MM can be associated with immune thrombocytopenic purpura (ITP), although this is rare. We present a case of MM with concurrent ITP which was refractory of steroids and intravenous immunoglobulin but had a response with anti-neoplastic therapy for MM. We also review all the cases of ITP with MM described in the literature. If conventional treatment for ITP associated with MM fails to improve platelet count, anti-neoplastic therapy for MM should be considered.


A systematic review and meta-analysis of studies evaluating the performance and operational characteristics of dual point-of-care tests for HIV and syphilis.
Mother-to-child transmission (MTCT) of syphilis and HIV continue to be important yet preventable causes of perinatal and infant morbidity and mortality. To systematically review, critically appraise and perform a meta-analysis to evaluate the operational characteristics of dual rapid diagnostic tests (RDTs) for HIV/syphilis and evaluate whether they are cost effective, acceptable and easy to use. Systematic review and meta-analysis. We searched seven electronic bibliographic databases from 2012 to December 2016 with no language restrictions. Search keywords included HIV, syphilis and diagnosis. We included studies that evaluated the operational characteristics of dual HIV/syphilis RDTs. Outcomes included diagnostic test accuracy, cost effectiveness, ease of use and interpretation and acceptability. All studies were assessed against quality criteria and assessed for risk of bias. Of 1914 identified papers, 18 were included for the meta-analysis of diagnostic accuracy for HIV and syphilis. All diagnostic accuracy evaluation studies showed a very high sensitivity and specificity for HIV and a lower, yet adequate, sensitivity and specificity for syphilis, with some variation among types of test. Dual screening for HIV and syphilis was more cost effective than single rapid tests for HIV and syphilis and prevented more adverse pregnancy outcomes. Qualitative data suggested dual RDTs were highly acceptable to clients, who cited time to result, cost and the requirement of a single finger prick as important characteristics of dual RDTs. The results of this systematic review and meta-analysis can be used by policy-makers and national programme managers who are considering implementing dual RDTs for HIV and syphilis. PROSPERO 2016:CRD42016049168.


Mechanical properties of the aorta determined by the pressure-diameter relation.
Elastic properties of the aorta represent an important determinant of left ventricular function and coronary blood flow. Aortic pressure-diameter relation provides insights into the mechanism, involved in the alterations of the elastic properties of the aorta. Recently, we described a new method to obtain aortic pressure-diameter relation in conscious humans. Aortic strain, distensibility, pressure-diameter relation, stiffness constant, augmentation index and aortic energy loss are indexes of aortic elastic properties derived by this method. Coronary artery disease is a potent independent factor associated with aortic elastic performance. Smoking produced an acute decrease of the elastic properties of the aorta in habitual smokers. Passive smoking has a similar unfavourable effect on aortic elastic properties. Elastic properties of the aorta are deteriorated in hypertensive patients compared to normotensive subjects and energy loss due to aortic wall viscosity is increased in the first group. Tachycardia induced by ventricular pacing produces an acute increase of the aortic distensibility in humans. Intravenous 17 beta-estradiol produces an improvement of the elastic properties of the aorta in menopausal women both with and without coronary artery disease. A decrease of aortic stiffness is demonstrated during intraaortic balloon pumping, associated with significant increase of the cardiac index and significant reduction of myocardial oxygen consumption. Thus, therapeutic interventions alter the elastic properties of the aorta and improvement of the elastic properties of the aorta may be beneficial in altering the natural history of diseases.


Leukocyte Associated Immunoglobulin Like Receptor 1 Regulation and Function on Monocytes and Dendritic Cells During Inflammation.
Inhibitory receptors are crucial immune regulators and are essential to prevent exacerbated responses, thus contributing to immune homeostasis. Leukocyte associated immunoglobulin like receptor 1 (LAIR-1) is an immune inhibitory receptor which has collagen and collagen domain containing proteins as ligands. LAIR-1 is broadly expressed on immune cells and has a large availability of ligands in both circulation and tissues, implicating a need for tight regulation of this interaction. In the current study, we sought to examine the regulation and function of LAIR-1 on monocyte, dendritic cell (DC) and macrophage subtypes, using different in vitro models. We found that LAIR-1 is highly expressed on intermediate monocytes as well as on plasmacytoid DCs. LAIR-1 is also expressed on skin immune cells, mainly on tissue CD14+ cells, macrophages and CD1c+ DCs. In vitro, monocyte and type-2 conventional DC stimulation leads to LAIR-1 upregulation, which may reflect the importance of LAIR-1 as negative regulator under inflammatory conditions. Indeed, we demonstrate that LAIR-1 ligation on monocytes inhibits toll like receptor (TLR)4 and Interferon (IFN)-α- induced signals. Furthermore, LAIR-1 is downregulated on GM-CSF and IFN-γ monocyte-derived macrophages and monocyte-derived DCs. In addition, LAIR-1 triggering during monocyte derived-DC differentiation results in significant phenotypic changes, as well as a different response to TLR4 and IFN-α stimulation. This indicates a role for LAIR-1 in skewing DC function, which impacts the cytokine expression profile of these cells. In conclusion, we demonstrate that LAIR-1 is consistently upregulated on monocytes and DC during the inflammatory phase of the immune response and tends to restore its expression during the resolution phase. Under inflammatory conditions, LAIR-1 has an inhibitory function, pointing toward to a potential intervention opportunity targeting LAIR-1 in inflammatory conditions.


Gut microbiota shape 'inflamm-ageing' cytokines and account for age-dependent decline in DNA damage repair.
Failing to properly repair damaged DNA drives the ageing process. Furthermore, age-related inflammation contributes to the manifestation of ageing. Recently, we demonstrated that the efficiency of repair of diethylnitrosamine (DEN)-induced double-strand breaks (DSBs) rapidly declines with age. We therefore hypothesised that with age, the decline in DNA damage repair stems from age-related inflammation. We used DEN-induced DNA damage in mouse livers and compared the efficiency of their resolution in different ages and following various permutations aimed at manipulating the liver age-related inflammation. We found that age-related deregulation of innate immunity was linked to altered gut microbiota. Consequently, antibiotic treatment, MyD88 ablation or germ-free mice had reduced cytokine expression and improved DSBs rejoining in 6-month-old mice. In contrast, feeding young mice with a high-fat diet enhanced inflammation and facilitated the decline in DSBs repair. This latter effect was reversed by antibiotic treatment. Kupffer cell replenishment or their inactivation with gadolinium chloride reduced proinflammatory cytokine expression and reversed the decline in DSBs repair. The addition of proinflammatory cytokines ablated DSBs rejoining mediated by macrophage-derived heparin-binding epidermal growth factor-like growth factor. Taken together, our results reveal a previously unrecognised link between commensal bacteria-induced inflammation that results in age-dependent decline in DNA damage repair. Importantly, the present study support the notion of a cell non-autonomous mechanism for age-related decline in DNA damage repair that is based on the presence of 'inflamm-ageing' cytokines in the tissue microenvironment, rather than an intrinsic cellular deficiency in the DNA repair machinery.


Epithelial re-growth is associated with inhibition of obliterative airway disease in orthotopic tracheal allografts in non-immunosuppressed rats.
Because epithelial cells are targets of alloimmune injury leading ultimately to airway obliteration, we tested whether epithelial re-growth could prevent obliterative airway disease (OAD) in orthotopic tracheal allografts. Brown Norway tracheal segments were orthotopically transplanted into nonimmunosuppressed Lewis rats. Allografts were removed on days 2-10 (n=13), 30 (n=4), and 60 (n=5) for histology, computerized morphometry (obliteration), and immunohistochemical detection of mononuclear cells, smooth muscle alpha-actin, and tissue phenotype. Normal tracheas, host tracheas, and heterotopically transplanted allografts served as controls. Orthotopic allografts removed on days 2-10 exhibited epithelial damage and re-growth and mononuclear cell infiltration. On days 30 and 60, partially ciliated cuboidal or attenuated epithelium completely covered the lumen. Although mononuclear cells declined, numerous T cells with a high CD4/CD8 ratio were found in the epithelium till day 60. Orthotopic allograft epithelium expressed donor phenotype on day 7, but recipient phenotype on days 30 and 60. Despite subepithelial alpha-actin positive myofibroblast proliferation, obliteration did not progress from day 7 to 30 and 60 (35, 30, and 33%, respectively). Although more than in normal or host tracheas, the obliteration in orthotopic allografts on days 30 and 60 was significantly less (P<0.001) than in heterotopic allografts. We describe, for the first time, longterm patency of fully histoincompatible orthotopic tracheal allografts in nonimmunosuppressed rats. Despite acute alloimmune injury and induction of myofibroblast proliferation, epithelial re-growth from the host limited the progression of OAD, thus emphasizing the role of epithelium in the control of airway obliteration.


Gene Expression Response to Sea Lice in Atlantic Salmon Skin: RNA Sequencing Comparison Between Resistant and Susceptible Animals.
Sea lice are parasitic copepods that cause large economic losses to salmon aquaculture worldwide. Frequent chemotherapeutic treatments are typically required to control this parasite, and alternative measures such as breeding for improved host resistance are desirable. Insight into the host-parasite interaction and mechanisms of host resistance can lead to improvements in selective breeding, and potentially novel treatment targets. In this study, RNA sequencing was used to study the skin transcriptome of Atlantic salmon (Salmo salar) parasitized with sea lice (Caligus rogercresseyi). The overall aims were to compare the transcriptomic profile of skin at louse attachment sites and "healthy" skin, and to assess differences in gene expression response between animals with varying levels of resistance to the parasite. Atlantic salmon pre-smolts were challenged with C. rogercresseyi, growth and lice count measurements were taken for each fish. 21 animals were selected and RNA-Seq was performed on skin from a louse attachment site, and skin distal to attachment sites for each animal. These animals were classified into family-balanced groups according to the traits of resistance (high vs. low lice count), and growth during infestation. Overall comparison of skin from louse attachment sites vs. healthy skin showed that 4,355 genes were differentially expressed, indicating local up-regulation of several immune pathways and activation of tissue repair mechanisms. Comparison between resistant and susceptible animals highlighted expression differences in several immune response and pattern recognition genes, and also myogenic and iron availability factors. Components of the pathways involved in differential response to sea lice may be targets for studies aimed at improved or novel treatment strategies, or to prioritize candidate functional polymorphisms to enhance genomic selection for host resistance in commercial salmon breeding programs.


Clinical Tips: The New Commission on Dietetic Registration/Professional Development Portfolio Process: You're in the Driver's Seat.
After more than 25 years of credentialing dietetic professionals, the Commission on Dietetic Registration is revising the format for logging continuing professional education. As of 2001, the new system will be phased in over 5 years. Each registered dietetic professional will now plan a course of continuing education to meet personal and professional growth needs. This is a very exciting process and one that has many asking questions of "why, how, when, and where." This article is designed to put a perspective on the new process.


Twisted bio-nanorods serve as a template for constructing chiroptically active nanoflowers.
Using biomacromolecule-based nanorods as a chiral source is a creative way to fabricate broadband chiroptically active nanoarchitectures. Herein, right-handedly twisted cellulose nanocrystal (CNC) nanorods, serving simultaneously as a chiral template and a building block, were combined with CuO for constructing composite nanoflowers that showed broadband chiroptical activity. Through calcination, the CuO/CNC nanoflowers can transform into chiral CuO with a flower-like architecture. The established preparation technology is expected to provide various novel broadband chiroptically active nanoarchitectures originating from CNCs and metal oxides.


Familial early puberty: presentation and inheritance pattern in 139 families.
The mechanism that initiates the onset of puberty is largely unknown but the age of onset is mainly under genetic control and influenced by environmental factors including nutrition. The coexistence in the same family of central precocious puberty and advanced puberty, both representing early puberty, suggests that they may represent a clinical spectrum of the same trait due to early activation of the GnRH pulse generator. We therefore evaluated the mode of inheritance of early puberty in a large series of familial cases. A retrospective, single center study was carried out on 154 probands (116 girls and 38 boys), from 139 families seen for idiopathic central precocious puberty (onset before 8 years in girls and 9-10 years in boys, n = 93) and/or advanced puberty (onset between 8 and 10 years in girls and 10 and 11 years in boys, n = 61) seen over a period of 8 years. Of the 139 families, 111 (80.4 %) had at least one affected 1st degree relatives, 17 (12 %) had only 2nd, 5 (3.6 %) only 3rd and 3 (2.2 %) had both 2nd and 3rd degree affected individuals. In the two remaining families, the unaffected mother had affected girls from two unaffected fathers. In the majority of families the inheritance of the phenotype was consistent with autosomal dominant mode of transmission with incomplete penetrance. An exclusively maternal mode of transmission could be observed or inferred in 83 families, paternal in only 2 families (p < 0.0001) and both maternal and paternal modes in 15 families. In the 139 families, 374 cases of early puberty were identified of whom 315 (84.2 %) were affected females and 59 (15.8 %) affected males (p < 0.0001). Twenty one percent of families had exclusively precocious puberty, 25 % had exclusively advanced puberty and 54 % had combinations of both. The data confirm the high incidence of affected girls with familial early puberty. The mode of inheritance of the phenotype is predominantly maternal. More than half of the families included both precocious and advanced puberty suggesting similar genetic factors.


Does a lack of auditory experience affect sequential learning?
To understand the interaction between sensory experiences and cognition, it is critical to investigate the possibility that deprivation in one sensory modality might affect cognition in other modalities. Here we are concerned with the hypothesis that early experience with sound is vital to the development of domain-general sequential processing skills. In line with this hypothesis, a seminal empirical study found that prelingually deaf children had impaired sequence learning in the visual modality. In order to assess the limits of this hypothesis, the current study employed a different visual sequence learning task in an investigation of prelingually deaf children with cochlear implants and normal hearing children. Results showed statistically significant learning in each of the two groups, and no significant difference in the amount of learning between groups. Moreover, there was no association between the age at which the child received their implant (and thus access to electric hearing) and their performance on the sequential learning task. We discuss key differences between our study and the previous study, and argue that the field must reconsider claims about domain-general cognitive impairment resulting from early auditory deprivation.


Comparative analysis of peri-implant marginal bone loss based on microthread location: a 1-year prospective study after loading.
The purpose of the present study was to investigate the short-term effects of microthread location on peri-implant marginal bone levels. Two types of implants, one with microthreads placed at the implant top (group A) and the other with microthreads placed 0.5 mm below the implant top (group B), were placed adjacent to each other in the partially edentulous areas of 20 patients. In total, 40 implants were placed. Bone loss around each implant was analyzed after 1 year of functional loading, and gingival parameters (modified plaque index and modified sulcus bleeding index) of the peri-implant soft tissue were evaluated. Bone losses after loading and gingival parameters were compared using the paired t test. The average bone loss was 0.16 (SD: 0.19) mm in group A and 0.30 (SD: 0.22) mm in group B after 1-year of functional loading. The paired t test revealed a significant difference in crestal bone loss between groups A and B in individual patients (P = 0.004). No significant differences were found between the two groups for the gingival parameters. Less peri-implant bone loss was observed around implants with microthreads placed at the implant top (group A) compared to those in which microthreads were placed below the top (group B). These results indicated that the microthreads acted to stabilize the peri-implant marginal bone, and their locations played an important role in the stabilization process.


Colour constancy under simultaneous changes in surface position and illuminant.
Two kinds of constancy underlie the everyday perception of surface colour: constancy under changes in illuminant and constancy under changes in surface position. Classically, these two constancies seem to place conflicting demands on the visual system: to both take into account the region surrounding a surface and also discount it. It is shown here, however, that the ability of observers to make surface-colour matches across simultaneous changes in test-surface position and illuminant in computer-generated 'Mondrian' patterns is almost as good as across changes in illuminant alone. Performance was no poorer when the surfaces surrounding the test surface were permuted, or when information from a potential comparison surface, the one with the highest luminance, was suppressed. Computer simulations of cone-photoreceptor activity showed that a reliable cue for making surface-colour matches in all experimental conditions was provided by the ratios of cone excitations between the test surfaces and a spatial average over the whole pattern.


Fasciotomy, chronic venous insufficiency, and the calf muscle pump.
To test the hypothesis that fasciotomy may impair the function of the calf muscle pump, which in turn could result in the development of chronic venous insufficiency. A cohort study of patients with a history of lower extremity fasciotomy. An urban trauma center. Seventeen of the 83 patients identified through trauma, vascular, and/or orthopedic registries consented to participation in this study. Participating patients completed a study questionnaire, and then underwent a complete vascular examination, including air plethysmographic (APG) assessment. Patients with a history of venous injuries were also studied with color flow duplex venous imaging. Function of the calf muscle pump as measured by APG, and evidence of chronic venous insufficiency as measured by APG, findings on clinical examination, and by venous ultrasonography. Seventeen patients completed the study, including 8 with a history of vascular injuries, 6 with old fractures, and 3 who had undergone fasciotomy for soft tissue infections. The time from injury to examination ranged from 5 months to 20 years. Eight patients had signs or symptoms of venous insufficiency, the severity of which appeared to be time dependent. The APG data showed significant mean differences between fasciotomy and control extremities in ejection fraction (P<.001) and residual volume fraction (P<.001), both measures of calf muscle pump function. There were no significant changes in venous filling index, a measure of venous reflux, or in outflow fraction, which correlates with venous obstruction. There were no differences in APG variables between patients with vascular injuries vs those with orthopedic or soft tissue injuries. Lower extremity fasciotomy impairs long-term calf muscle pump function, as measured by APG, in patients with and without vascular injuries. These patients are at risk for the long-term development of chronic venous insufficiency following lower extremity trauma.


Experimental argon laser photocoagulation. II. Effects on the optic disc.
Eight monkeys and five humans were subjected to photocoagulation with single argon laser burns of varying intensity on the optic nerve head. Energies greater than 400 milliwatts, 0.2 second and 100 mu spot size consistently caused neural parenchymal damage, even in the absence of heat-generating pigment epithelium. Papillary burns in monkeys and humans created with powers less than 400 mW showed only small, focal submicroscopic areas of degeneration, and were therefore considered relatively safe except for the occasional involvement of critical foveal fibers in which scotomas of disproportionately greater severity than one would expect from the size of the lesion could result. The threshold for toxicity following peripapillary photocoagulation is lower, because the burned pigment epithelium can radiate heat into the adjacent optic nerve fibers. Normal capillaries within the optic nerve were seldom destroyed or occluded, even after high energy densities. In normal vessels, we have found it impossible to create a highly selective lesion without concurrently causing neural damage about the vessel.


Neonatal hyperoxia causes pulmonary vascular disease and shortens life span in aging mice.
Bronchopulmonary dysplasia is a chronic lung disease observed in premature infants requiring oxygen supplementation and ventilation. Although the use of exogenous surfactant and protective ventilation strategies has improved survival, the long-term pulmonary consequences of neonatal hyperoxia are unknown. Here, we investigate whether neonatal hyperoxia alters pulmonary function in aging mice. By 67 weeks of age, mice exposed to 100% oxygen between postnatal days 1 to 4 showed significantly a shortened life span (56.6% survival, n = 53) compared to siblings exposed to room air as neonates (100% survival, n = 47). Survivors had increased lung compliance and decreased elastance. There was also right ventricular hypertrophy and pathological evidence for pulmonary hypertension, defined by reduction of the distal microvasculature and the presence of numerous dilated arterioles expressing von Willebrand factor and α-smooth muscle actin. Consistent with recent literature implicating bone morphogenetic protein (BMP) signaling in pulmonary vascular disease, BMP receptors and downstream phospho-Smad1/5/8 were reduced in lungs of aging mice exposed to neonatal oxygen. BMP signaling alterations were not observed in 8-week-old mice. These data suggest that loss of BMP signaling in aged mice exposed to neonatal oxygen is associated with a shortened life span, pulmonary vascular disease, and associated cardiac failure. People exposed to hyperoxia as neonates may be at increased risk for pulmonary hypertension.


Beta 2-adrenoceptors mediate the stimulating effect of adrenaline on active electrogenic Na-K-transport in rat soleus muscle.
The relative role of beta 1- and beta 2-adrenoceptors in mediating the stimulating effect of adrenaline on active electrogenic Na-K-transport has been assessed in experiments on rat soleus muscles in vitro and in vivo. 2 In the rat isolated soleus muscle, adrenaline (10(-6) M) increases the resting membrane potential (EM) by 5.8 mV and stimulates 22Na-efflux and ouabain-suppressible 42K-uptake by 91 and 94%, respectively. 3 All of these effects are completely blocked by propranolol (10(-5) M), whereas the beta 1-selective adrenoceptor antagonist, metoprolol, was found to be at least 50 times less potent. 4 The beta 2-adrenoceptor agonist, salbutamol, was at least 100 times as potent as H133/22 (a beta 1-selective agonist) in stimulating 22Na-efflux and 42K-influx. 5 In experiments performed under pentobarbitone anaesthesia, the intravenous injection of adrenaline (5 microgram) or salbutamol (0.5 to 50 microgram) led to a rapid and marked increase in the EM of the exposed soleus muscle. This hyperpolarizing effect could not be accounted for by the concomitant, relatively modest change in extracellular K.


The character of gene expression of human periosteum used to form new tissue in allograft bone.
Of more than 2 million segmental bone defects repaired annually with bone autografts and allografts, 15-40% fail. Improving healing rates may be approached with tissue engineering and use of periosteum overlying an allograft. The present study documents gene expression in human periosteum-allograft constructs compared to allografts alone. Strips of human cadaveric periosteum (26 years, f, distal femur) were sutured about sterilized human femoral cortical strut bone allograft (54 years, m) segments. After construct incubation (M199 supplemented medium) for 8 d, constructs and allografts alone were implanted in nude mice. At 10 and 20 weeks, constructs (N = 4, each group) and allografts (N = 2, each group) were retrieved and placed in RNAlater for quantitative PCR to determine expression of human- and murine-specific genes relevant to remodeling. Specimens were frozen-ground to powders and RNA was extracted, purified, reverse-transcribed, and amplified. Ribosomal protein (P0) was used to normalize sample quantities. Fold change plots were generated following statistical analyses comparing 20- to 10-week gene expression data. Allografts alone yielded no human-specific gene expression. Notable fold changes of human-specific alkaline phosphatase, bone sialoprotein, type I collagen, decorin, RANKL, RANK, cathepsin K, and osteocalcin in 20-week compared to 10-week specimens were found. Murine-specific expression of genes indicative of host mouse vascularization (RANK, type I collagen) was detected in both allograft alone and periosteum-allograft samples. Gene data confirm viable periosteum in constructs after 20 weeks. Relatively higher fold-change values of RANK, RANKL and cathepsin K indicate activities of osteoclast precursors, osteoclasts and osteoblasts involved in allograft remodeling during implantation. All additional genes of interest indicate osteoblast activity in new bone matrix formation. Gene data are directly correlated with previous and present histology work. The results of this study suggest that further investigations could help to establish whether autologous periosteum-allograft constructs could be used for the repair of bone defects.


The adjuvant mechanism of cationic dimethyldioctadecylammonium liposomes.
Cationic liposomes are being used increasingly as efficient adjuvants for subunit vaccines but their precise mechanism of action is still unknown. Here, we investigated the adjuvant mechanism of cationic liposomes based on the synthetic amphiphile dimethyldioctadecylammonium (DDA). The liposomes did not have an effect on the maturation of murine bone-marrow-derived dendritic cells (BM-DCs) related to the surface expression of major histocompatibility complex (MHC) class II, CD40, CD80 and CD86. We found that ovalbumin (OVA) readily associated with the liposomes (> 90%) when mixed in equal concentrations. This efficient adsorption onto the liposomes led to an enhanced uptake of OVA by BM-DCs as assessed by flow cytometry and confocal fluorescence laser-scanning microscopy. This was an active process, which was arrested at 4 degrees and by an inhibitor of actin-dependent endocytosis, cytochalasin D. In vivo studies confirmed the observed effect because adsorption of OVA onto DDA liposomes enhanced the uptake of the antigen by peritoneal exudate cells after intraperitoneal injection. The liposomes targeted antigen preferentially to antigen-presenting cells because we only observed a minimal uptake by T cells in mixed splenocyte cultures. The adsorption of antigen onto the liposomes increased the efficiency of antigen presentation more than 100 times in a responder assay with MHC class II-restricted OVA-specific T-cell receptor transgenic DO11.10 T cells. Our data therefore suggest that the primary adjuvant mechanism of cationic DDA liposomes is to target the cell membrane of antigen-presenting cells, which subsequently leads to enhanced uptake and presentation of antigen.


Gene expression in normal human bronchial epithelial (NHBE) cells following in vitro exposure to cigarette smoke condensate.
Cigarettes that burn tobacco produce a complex mixture of chemicals, including mutagens and carcinogens. Cigarettes that primarily heat tobacco produce smoke with marked reductions in the amount of mutagens and carcinogens and demonstrate reduced mutagenicity and carcinogenicity in a battery of toxicological assays. Chemically induced oxidative stress, DNA damage, and inflammation may alter cell cycle regulation and are important biological events in the carcinogenic process. The objective of this study was to characterize and compare the effects of smoke condensates from cigarettes that burn tobacco and those that primarily heat tobacco on gene expression in NHBE cells. For this comparison, we used quantitative RT/PCR and further evaluated the effects on cell cycling using flow cytometry. Cigarette smoke condensates (CSCs) were prepared from Kentucky 1R4F cigarettes (a tobacco-burning product designed to represent the average full-flavor, low "tar" cigarette in the US market) and Eclipse (a cigarette that primarily heats tobacco) using FTC machine smoking conditions. The CSC from 1R4F cigarettes induced statistically significant increases in the mRNA levels of genes responsive to DNA damage (GADD45) and involved in cell cycle regulation (p21;WAF1/CIP1), compared to the CSC from Eclipse cigarettes. In addition, genes coding for cyclooxygenase-2 (COX-2) and interleukin 8 (IL-8), which are associated with oxidative stress and inflammation, respectively, were increased statistically significantly more by CSC from 1R4F than by that from Eclipse. Furthermore, a dose-dependent increase in IL-8 protein secretion into cell culture media was stimulated by 1R4F exposure, whereas minimal IL-8 protein was secreted after Eclipse treatment. The biological relevance of the differential effect on gene expression was reflected in differential cell cycle regulation, as cells exposed to 1R4F CSC exhibited more significant S phase and G2 phase accumulation than cells exposed to Eclipse CSC. These data indicate that the simplified smoke chemistry of the tobacco-heating Eclipse cigarette yields statistically significant reductions in the expression of key genes involved in DNA damage, oxidative stress, inflammatory response, and cell cycle regulation in normal human bronchial epithelial cells compared to a representative tobacco-burning cigarette.


Preventing sexually transmissible infections in Australian general practice.
The aims of the present study were to explore aspects of sexually transmissible infections (STI) prevention in general practice and to examine general practitioners' (GPs) perceived barriers to sexual-health promotion. The data from a postal survey of 409 GPs practising in New South Wales, Australia (response rate 45.4%) are analysed to explore the prevention of STI in general practice and to examine practitioners' perceived barriers to sexual-health promotion. About 49% of GPs reported having STI leaflets/pamphlets for patients in their clinic, while 21% had posters on STI displayed in their waiting room. Two-third (67%) of GPs provided STI-specific printed materials/leaflets to patients with STI. Female GPs were more likely to be proactive in STI prevention. Time and funding appear to be the major barriers to sexual-health promotion, followed by inadequate access to counselling. One-fifth (22%) of GPs felt that they had little influence in changing patients' risk behaviour, while about 28% emphasized the need for further preventive care training. The present study identifies inconsistencies in STI-prevention activities in general practice along with barriers to undertake sexual-health promotion. This area warrants further attention if GPs are to contribute fully to the control of STI.


Dietary exposure to persistent organic pollutants and metals among Inuit and Chukchi in Russian Arctic Chukotka.
The general aim was to assess dietary exposure to selected persistent organic pollutants (POPs) and metals among Eskimo (Inuit) and Chukchi of the Chukotka Peninsula of the Russian Arctic, and to establish recommendations for exposure risk reduction. A cross-sectional evaluation of nutritional patterns of coastal and inland indigenous peoples of the Chukotka Autonomous Okrug (in 2001-2003); assessment of the levels of persistent toxic substances (PTSs) in traditional foods and their comparison to Russian food safety limits; the identification of local sources of food contamination; and the recommendation and implementation of risk management measures. Community-based dietary survey of self reported food frequencies (453 persons), chemical analyses (POPs and metals) of local foods and indoor matters (397 samples), substantiation of recommendations for daily (weekly, monthly) intakes of traditional food. POPs in traditional food items are generally below the Russian food safety limits except marine mammal fat, while Hg and Cd are high mainly in mammal viscera. Lead is relatively low in tissues of all animals studied. For the Chukotka coastal communities, seals constitute the principal source of the whole suite of PTSs considered. Consumption restrictions are recommended for marine and freshwater fish, some wild meats (waterfowl and seal), fats (whale and seal), liver (most animals) and kidney (reindeer, walrus and seal). Evidence is presented that contamination of foodstuffs may be significantly increased during storing/processing/cooking of food due to indoor and outdoor environmental conditions. Based on the analytical findings and the local PTSs sources identified, guidelines on food safety are suggested, as well as measures to reduce food contamination and domestic and local sources. Important and urgent remedial actions are recommended to minimize PTSs environmental and domestic contamination. Waste clean-up activities started in coastal Chukotka in 2007.


Sequence and structural elements that contribute to efficient encephalomyocarditis virus RNA translation.
The nucleotide sequence of the 5' nontranslated region of encephalomyocarditis virus (EMCV-Rueckert) was determined, and a consensus RNA structural model for this sequence (850 bases) and three other poly(C)-containing cardioviruses (mengovirus, EMCV-B, and EMCV-D) was created through reiterative use of a minimum-free-energy folding algorithm. The RNA elements within this region which contribute to translation of EMCV proteins were mapped in cell-free reactions programmed with cDNA-derived RNA transcripts. The data provide evidence that stem-loop motifs I, J and K, formed by viral bases 451 to 785, are important components of cap-independent translation. In contrast to other reports, a minimal role for stem-loop H (bases 406 to 444) in translational activity is indicated. Small 5' nontranslated region fragments (bases 667 to 797) containing the J and K motifs proved strong competitive inhibitors when added to cell-free reactions programmed with exogenous capped or uncapped mRNAs. The putative sequestering of required translational factors by this segment clearly contributes to translational activity, but also suggests a possible competitive mechanism for the down regulation of host protein synthesis during viral infection.