Sample size calculations for disease freedom and prevalence estimation surveys.
We developed a Bayesian approach to sample size calculations for studies designed to estimate disease prevalence that uses a hierarchical model for estimating the proportion of infected clusters (cluster-level prevalence) within a country or region. The clusters may, for instance, be villages within a region, cities within a state, or herds within a country. Our model allows for clusters with zero prevalence and for variability in prevalences among infected clusters. Moreover, uncertainty about diagnostic test accuracy and within-cluster prevalences is accounted for in the model. A predictive approach is used to address the issue of sample size selection in human and animal health surveys. We present sample size calculations for surveys designed to substantiate freedom of a region from an infectious agent (disease freedom surveys) and for surveys designed to estimate cluster-level prevalence of an endemic disease (prevalence estimation surveys). In disease freedom surveys, for instance, assuming the cluster-level prevalence for a particular infectious agent in the region is greater than a maximum acceptable threshold, a sample size combination consisting of the number of clusters sampled and number of subjects sampled per cluster can be determined for which authorities conducting the survey detect this excessive cluster-level prevalence with high predictive probability. The method is straightforward to implement using the Splus/R library emBedBUGS together with WinBUGS.


Impact of wind turbine noise in the Netherlands.
The Dutch government aims at an increase of wind energy up to 6 000 MW in 2020 by placing new wind turbines on land or offshore. At the same time, the existing noise legislation for wind turbines is being reconsidered. For the purpose of establishing a new noise reception limit value expressed in L den , the impact of wind turbine noise under the given policy targets needs to be explored. For this purpose, the consequences of different reception limit values for the new Dutch noise legislation have been studied, both in terms of effects on the population and regarding sustainable energy policy targets. On the basis of a nation-wide noise map containing all wind turbines in The Netherlands, it is calculated that 3% of the inhabitants of The Netherlands are currently exposed to noise from wind turbines above 28 dB(A) at the faηade. Newly established dose-response relationships indicate that about 1500 of these inhabitants are likely to be severely annoyed inside their dwellings. The available space for new wind turbines strongly depends on the noise limit value that will be chosen. This study suggests an outdoor A-weighted reception limit of L den = 45 dB as a trade-off between the need for protection against noise annoyance and the feasibility of national targets for renewable energy.


Parents' Perceptions about Salt Consumption in Urban Areas of Peru: Formative Research for a Social Marketing Strategy.
Salt intakes in Latin America currently double the World Health Organization's recommendation of 5 g/day. Various strategies to reduce the population's salt consumption, such as raising awareness using social marketing, have been recommended. This study identified parents' perceptions of salt consumption to inform a social marketing strategy focused on urban areas in Peru. Using a sequential exploratory methods design, parents of pre-school children, of high and low socioeconomic status, provided qualitative data in the form of interviews and focus groups. Following this, quantitative data was obtained via questionnaires, which were sent to all parents. The information was analyzed jointly. 296 people (mean age 35.4, 82% women) participated, 64 in the qualitative and 232 in the quantitative phase of the study. Qualitative data from the first phase revealed that the majority of mothers were in charge of cooking, and female participants expressed that cooking was "their duty" as housewives. The qualitative phase also revealed that despite the majority of the participants considered their salt intake as adequate, half of them mentioned that they have tried to reduce salt consumption, and the change in the flavor of the food was stated as the most difficult challenge to continue with such practice. Quantitative data showed that 67% of participants would be willing to reduce their salt intake, and 79.7% recognized that high salt intake causes hypertension. In total, 84% of participants reaffirmed that mothers were in charge of cooking. There were no salient differences in terms of responses provided by participants from high versus low socioeconomic groups. The results point towards the identification of women as a potential target-audience of a social marketing strategy to promote reductions in salt intake in their families and, therefore, a gender-responsive social marketing intervention is recommended.


Virtual reconstruction and geometric morphometrics as tools for paleopathology: a new approach to study rare developmental disorders of the skeleton.
Survey studies of osteoarchaeological collections occasionally yield specimens exhibiting rare skeletal developmental disorders. Beyond paleopathological diagnosis, however, it is often difficult to gain insight into the processes, mechanisms, and consequences of the pathology, notably because archaeological specimens are often fragmentary. Here, we propose a combination of virtual reconstruction (VR) and geometric morphometrics (GM) to address these issues. As an example, we use VR to reconstruct the only known archaeological specimen exhibiting persistence of the pelvic triradiate cartilage and compare it via GM with a set of healthy pelvises representing both sexes and different ontogenetic stages. Our results evidence (i) a marked deviation of the pathological pelvis from the adult mean shape, (ii) the retention of typical male features, and (iii) the retention of a paedomorphic ratio between iliac and ischiopubic size. Altogether, such data offer new insights into the modularity and integration of pelvic ontogeny, while at the same time demonstrating the usefulness of a combined VR/GM approach as complement to classical methods of paleopathology.


Identification of Candida dubliniensis among oral yeast isolates from an Italian population of human immunodeficiency virus-infected (HIV+) subjects.
Candida dubliniensis, an emerging oral pathogen, phenotypically resembles Candida albicans so closely that it is easily misidentified as such. The aim of the present study was to evaluate the usefulness of two phenotypic methods, growth at 45 degrees C and 2,3,5-triphenyltetrazolium chloride (TTC) reduction, for confirming presumptive identification of C. dubliniensis and C. albicans by colony color on CHROMagar Candida (CAC) medium. A combination of these methods was used to establish the prevalence of oral C. dubliniensis in an Italian population of 45 human immunodeficiency virus (HIV)-infected subjects. Twenty-two samples (48.9%) were positive for yeasts on CAC medium producing a total of 37 fungal isolates. The colony color and 45 degrees C growth ability test correctly identified all C. dubliniensis and C. albicans isolates (5/37, 13.5%, and 16/37, 43.2%, respectively), while assessment of TTC reduction misidentified one C. albicans isolate. The isolation rate of C. dubliniensis was 11.1% (5/45 patients). All of the C. dubliniensis isolates were highly susceptible to fluconazole (MIC = 0.5 microg/ml). The combination of CAC medium screening with growth at 45 degrees C and TTC reduction tests may represent a simple, reliable and inexpensive identification protocol for C. dubliniensis.


Transfer Function Analysis of Dynamic Blood Flow Control in the Rat Kidney.
Renal blood flow is regulated by the myogenic response (MR) and tubuloglomerular feedback (TGF). Both mechanisms function to buffer not only steady pressure perturbations but also transient ones. In this study, we develop two models of renal autoregulation-a comprehensive model and a simplified model-and use them to analyze the individual contributions of MR and TGF in buffering transient pressure perturbations. Both models represent a single nephron of a rat kidney together with the associated vasculature. The comprehensive model includes detailed representation of the vascular properties and cellular processes. In contrast, the simplified model represents a minimal set of key processes. To assess the degree to which fluctuations in renal perfusion pressure at different frequencies are attenuated, we derive a transfer function for each model. The transfer functions of both models predict resonance at 45 and 180 mHz, which are associated with TGF and MR, respectively, effective autoregulation below [Formula: see text]100 mHz, and amplification of pressure perturbations above [Formula: see text]200 mHz. The predictions are in good agreement with experimental findings.


Staphylococcus aureus Community-acquired Pneumonia: Prevalence, Clinical Characteristics, and Outcomes.
Prevalence of Staphylococcus aureus community-acquired pneumonia (CAP) and its clinical features remain incompletely understood, complicating empirical selection of antibiotics. Using a multicenter, prospective surveillance study of adults hospitalized with CAP, we calculated the prevalence of methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) among all CAP episodes. We compared the epidemiologic, radiographic, and clinical characteristics of S. aureus CAP (per respiratory or blood culture) with those of pneumococcal (per respiratory or blood culture or urine antigen) and all-cause non-S. aureus CAP using descriptive statistics. Among 2259 adults hospitalized for CAP, 37 (1.6%) had S. aureus identified, including 15 (0.7%) with MRSA and 22 (1.0%) with MSSA; 115 (5.1%) had Streptococcus pneumoniae Vancomycin or linezolid was administered to 674 (29.8%) patients within the first 3 days of hospitalization. Chronic hemodialysis use was more common among patients with MRSA (20.0%) than pneumococcal (2.6%) and all-cause non-S. aureus (3.7%) CAP. Otherwise, clinical features at admission were similar, including concurrent influenza infection, hemoptysis, multilobar infiltrates, and prehospital antibiotics. Patients with MRSA CAP had more severe clinical outcomes than those with pneumococcal CAP, including intensive care unit admission (86.7% vs 34.8%) and in-patient mortality (13.3% vs 4.4%). Despite very low prevalence of S. aureus and, specifically, MRSA, nearly one-third of adults hospitalized with CAP received anti-MRSA antibiotics. The clinical presentation of MRSA CAP overlapped substantially with pneumococcal CAP, highlighting the challenge of accurately targeting empirical anti-MRSA antibiotics with currently available clinical tools and the need for new diagnostic strategies.


Tuning the separation and coupling of corannulene trianion-radicals through sizable alkali metal belts.
The first heterobimetallic sandwich-type aggregate formed by bowl-shaped corannulene trianion-radicals, C20H10˙3-, has been synthesized using mixed-metal reduction of C20H10. The product was crystallographically characterized to reveal the self-assembly of [Cs+//(C20H103-)/4K+/(C20H103-)//Cs+], in which two triply-charged corannulene decks encapsulate a rectangle of four potassium ions (the K···K separations are 4.212(4) and 5.185(4) Å), with the exterior concave bowl cavities being selectively filled by one cesium ion each. In order to provide insights into the geometrical features and electronic structure of this novel mixed-metal organometallic self-assembly, an in-depth theoretical investigation has been carried out. Specifically, the influence of internal metal binding on the geometry and magnetic coupling of C20H10˙3- radicals is investigated for Group 1 metals. This study reveals that replacement of the sandwiched potassium ions with larger (Cs) and smaller (Li) ions allows variation of the size of the encapsulated metal belts, and thus enables tuning of the coupling of C20H10˙3- radicals.


The bacterial toxin toolkit.
Pathogenic bacteria and higher eukaryotes have spent a long time together, leading to a precise understanding of one another's way of functioning. Through rapid evolution, bacteria have engineered increasingly sophisticated weapons to hit exactly where it hurts, interfering with fundamental host functions. However, toxins are not only useful to the bacteria - they have also become an essential asset for life scientists, who can now use them as toolkits to explore cellular processes.


Student acceptance of e-learning methods in the laboratory class in Optometry.
Today's students have increased expectations for flexible learning options and evidence-based practice resources to be available to support curricular activities. We investigated: (i) the suitability of a static website for teaching ocular anatomy and physiology and an interactive version of the website with quiz and self-assessment activities and (ii) the usefulness of a blended online and in-lab environment to teach in Optometry. We administered a survey to compare responses of optometry students who had access to the interactive website, with those from students from a previous year who used the static version. We examined learning preferences of students in a focus group. Students were positive about the value of the website for their learning and the clarity of the website content. Nevertheless, objective comparison of pass rates for students using the static and interactive websites did not show significant changes. The majority of students commenting on the static website felt they did not get sufficient feedback via the website (67%) compared with only 22% from students who used self-assessments in the interactive website. Interestingly, users of the static website commented that it was perceived as just another resource while users of the interactive website commented on the usefulness of the material to review knowledge before laboratories. In the focus group, students reported they preferred a blended learning over the website alone even by students using the interactive website as they felt the need to revise content with the educator before the test. We conclude that there is acceptance of online learning methods due to the technologically 'savvy' environment of students in the first year of the Optometry programme but there is still dependence on the educator as the main administrator of their learning.


International funding for mental health: a review of the last decade.
Mental health represents one of the most significant and increasing burdens to global public health. Over the past decade, the once invisible field has gained recognition on the global health agenda and this increased recognition is expected to increase international funding for mental health. Our review found that few studies have been conducted as to the level of international funding for mental health and there is a need for a differentiated assessment. We conducted such an assessment of global development-related assistance for mental health between 2006 and 2016 and established categories to serve as a baseline for future measurement. We found that development assistance specifically dedicated to mental health accounted for just 0.3% of all development assistance for health. Given the limited public expenditure on mental health by national governments in low- and middle-income countries, renewed efforts by the international community and development partners to substantially increase funding for mental health are critical.


Childhood Dietary Intake in Italy: The Epidemiological "MY FOOD DIARY" Survey.
Promoting a healthy lifestyle during the first years of life is a key strategy for controlling obesity risk in later life; having good-quality epidemiological data on eating habits of infants and toddlers can improve awareness and possibly the education given by pediatricians to parents and children. With this aim, we performed a survey about the dietary pattern of Italian children in early childhood. We described the intake of energy, macronutrients and fiber, minerals, and vitamins of 443 Italian children (range 6.4-131 months), through a three-day food record filled out by their parents and assessed by family pediatricians. The results were compared with the Italian Dietary Reference Values. The median protein intake, in g/kg per body weight, exceeded the average requirement in all age groups, and in the 12-36 month period, the intake as % of energy was outside the reference range (>15%). The majority of the children consumed quantities of simple carbohydrates (consisting of both natural sugars and free or added sugars, 82.3% of the children in the study) and saturated fats (69% of the children in the study) above the limits of the Italian Dietary Reference Values, with low intake of fiber and polyunsaturated fats. Median mineral intake, in our study, was different depending on age, while vitamin D intake was very low in all age groups. This is one of the few studies reporting on the nutrient intake of Italian children with reference to nutrition recommendations in order to identify the principal nutritional errors. The present results underline the need for healthcare policies starting from the first years of life in order to ameliorate nutrient intake during childhood, possibly impacting long-term health outcomes.


Cortisol-mediated potentiation of uterine artery contractility: effect of pregnancy.
During pregnancy, maternal plasma cortisol concentrations approximately double. Given that cortisol plays an important role in the regulation of vascular reactivity, the present study investigated the potential role of cortisol in potentiation of uterine artery (UA) contractility and tested the hypothesis that pregnancy downregulated the cortisol-mediated potentiation. In vitro cortisol treatment (3, 10, or 30 ng/ml for 24 h) produced a dose-dependent increase in norepinephrine (NE)-induced contractions in both nonpregnant and pregnant (138-143 days gestation) sheep UA. However, this cortisol-mediated response was significantly attenuated by approximately 50% in pregnant UA. The 11 beta-hydroxysteroid dehydrogenase (11-beta HSD) inhibitor carbenoxolone did not change the effect of cortisol in nonpregnant UA but abolished its effect in pregnant UA by increasing the NE pD(2) in control tissues from 6.20 +/- 0.05 to 6.59 +/- 0.11. The apparent dissociation constant value of NE alpha(1)-adrenoceptors was not changed by cortisol in pregnant UA but was decreased in nonpregnant UA. There was no difference in glucocorticoid receptor density between nonpregnant and pregnant UA. Cortisol significantly decreased endothelial nitric oxide (NO) synthase protein levels and NO release in both nonpregnant and pregnant UA, but the effect of cortisol was attenuated in pregnant UA by approximately 50%. Carbenoxolone alone had no effects on NO release in nonpregnant UA but was decreased in pregnant UA. These results suggest that cortisol potentiates NE-mediated contractions by decreasing NO release and increasing NE-binding affinity to alpha(1)-adrenoceptors in nonpregnant UA. Pregnancy attenuates UA sensitivity to cortisol, which may be mediated by increasing type-2 11-beta HSD activity in UA.


Electrical Impedance Measurements of Biological Cells in Response to External Stimuli.
Dielectric spectroscopy (DS) is a noninvasive technique for real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for capturing, DS measurements, and unloading of biological cells. Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. A human prostate cancer cell line, PC-3, was used to evaluate the device performance. Suspension of PC-3 cells in low conductivity buffers (LCB) enhanced their dielectrophoretic trapping and impedance response. We report the time course of the variations in dielectric properties of PC-3 cells suspended in LCB and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. Importantly, we demonstrated that our device enabled real-time measurements of dielectric properties of live cancer cells and allowed the assessment of the cellular response to variations in buffer conductivity and pH. These data support further development of this device toward single cell measurements.


Chewing cycles in 4- and 5-year-old Down's syndrome children: a comparison of eating efficacy with normals.
Chewing movements of Down's syndrome children were measured and compared with those of normal preschool children. Twenty-six Down's syndrome children were monitored: 14 were 4 (8 males, 6 females) and 12 were 5 years old (6 males, 6 females). Chewing movements were measured by time (in seconds), number of cycles, and a time/cycle ratio. A chewing cycle was defined as an upward and downward movement of the chin. The time/cycle ratio is the total time, from the moment food was placed in the mouth until the final swallow occurred, divided by the number of cycles counted for the same period. The children's ages and gender did not affect time, cycles, or the time/cycle ratio. However, the measures were strongly affected by the type of food eaten. Children with Down's syndrome chew at a rate comparable to that of normal children. However, the duration of chewing is significantly prolonged per bite of food. This may be attributed to these children's lack of chewing vigor or their inability, at the ages studied, to chew solid foods. Time/cycle ratios are an excellent index to document such differences. How much sensory differences or deficits contribute to these findings is not known.


The In Vitro Stability of Circulating Tumour DNA.
DNA from apoptotic cancer cells, present in the circulation, has the potential to facilitate genomic profiling and disease monitoring. However, only low fractions of total cell-free DNA originates from cancer cells, limiting the applicability of circulating tumour DNA (ctDNA). Optimal sample processing is consequently of uttermost importance. Therefore, we evaluated the in vitro stability of ctDNA. Blood was collected in 10 ml EDTA or Streck tubes. Three conditions (EDTA and Streck tubes in room temperature, EDTA tubes at five degrees) and four time points (plasma harvested from blood aliquots of each 10 ml tube in a time series up to 24 h) were investigated. Each condition was evaluated in five metastatic prostate cancer patients. Subsequently, three additional patients were collected enabling investigation of the in vitro stability in EDTA tubes up to 48 h. The in vitro stability of ctDNA was interrogated by low-pass whole genome sequencing which allows for the identification of somatic copy-number alterations (CNAs). In silico simulations demonstrated that non-parametric testing could detect a 1% contamination by white blood cell DNA. Mutational profiling was performed by targeted, in-solution based hybridization capture and subsequent sequencing. The allelic fraction of individual mutations was used as an estimate of the in vitro stability. Somatic CNAs were detected in all patients. Surprisingly, the ctDNA levels at zero hours were not significantly different to 24 or 48 hour in vitro incubation in any investigated condition. Subsequently, mutational profiling corroborated the conclusions from the CNA analysis. The stability of ctDNA simplifies logistics without the requirement of immediate processing or applying fixatives to prevent white blood cell lysis.


Synthesis and receptor binding of IgG1 peptides derived from the IgG Fc region.
The IgG binding Fcgamma receptors (FcgammaRs) play a key role in defence against pathogens by linking humoral and cell-mediated immune responses. Impaired expression and/or function of FcgammaR may result in the development of pathological autoimmunity. Considering the functions of FcgammaRs, they are potential target molecules for drug design to aim at developing novel anti-inflammatory and immunomodulatory therapies. Previous data mostly obtained by X-ray analysis of ligand-receptor complexes indicate the profound role of the CH2 domain in binding to various FcgammaRs. Our aim was to localize linear segments, which are able to bind and also to modulate the function of the low affinity FcgammaRs, like FcgammaRIIb and FcgammaRIIIa. To this end a set of overlapping octapeptides was prepared corresponding to the 231-298 sequence of IgG1 CH2 domain and tested for binding to human recombinant soluble FcgammaRIIb. Based on these results, a second group of peptides was synthesized and their binding properties to recombinant soluble FcgammaRIIb, as well as to FcgammaRs expressed on the cell surface, was investigated. Here we report that peptide representing the Arg(255)-Ser(267) sequence of IgG1 is implicated in the binding to FcgammaRIIb. In addition we found that peptides corresponding to the Arg(255)-Ser(267), Lys(288)-Ser(298) or Pro(230)-Val(240) when presented in a multimeric form conjugated to branched chain polypeptide in uniformly oriented copies induced the release of TNFalpha, a pro-inflammatory cytokine from MonoMac monocyte cell line. These findings indicate that these conjugated peptides are able to cluster the activating FcgammaRs, and mediate FcgammaR dependent function. Peptide Arg(255)-Ser(267) can also be considered as a lead for further functional studies.


Cardiomyocyte-targeted and 17β-estradiol-loaded acoustic nanoprobes as a theranostic platform for cardiac hypertrophy.
Theranostic perfluorocarbon nanoprobes have recently attracted attention due to their fascinating versatility in integrating diagnostics and therapeutics into a single system. Furthermore, although 17β-estradiol (E2) is a potential anti-hypertrophic drug, it has severe non-specific adverse effects in various organs. Therefore, we have developed cardiomyocyte-targeted theranostic nanoprobes to achieve concurrent targeted imaging and treatment of cardiac hypertrophy. We had successfully synthesized E2-loaded, primary cardiomyocyte (PCM) specific peptide-conjugated nanoprobes with perfluorocarbon (PFP) as a core (PCM-E2/PFPs) and demonstrated their stability and homogeneity. In vitro and in vivo studies confirmed that when exposed to low-intensity focused ultrasound (LIFU), these versatile PCM-E2/PFPs can be used as an amplifiable imaging contrast agent. Furthermore, the significantly accelerated release of E2 enhanced the therapeutic efficacy of the drug and prevented systemic side effects. PCM-E2/PFPs + LIFU treatment also significantly increased cardiac targeting and circulation time. Further therapeutic evaluations showed that PCM-E2/PFPs + LIFU suppressed cardiac hypertrophy to a greater extent compared to other treatments, revealing high efficiency in cardiac-targeted delivery and effective cardioprotection. Our novel theranostic nanoplatform could serve as a potential theranostic vector for cardiac diseases.


Antibody Responses to SARS-CoV-2 in Patients With Novel Coronavirus Disease 2019.
The novel coronavirus SARS-CoV-2 is a newly emerging virus. The antibody response in infected patients remains largely unknown, and the clinical value of antibody testing has not been fully demonstrated. 173 patients with SARS-CoV-2 infection were enrolled. Their serial plasma samples (n = 535) collected during hospitalization were tested for total antibodies (Ab), IgM, and IgG against SARS-CoV-2. The dynamics of antibodies with disease progress were analyzed. Among 173 patients, the seroconversion rates for Ab, IgM, and IgG were 93.1%, 82.7%, and 64.7%, respectively. The reason for the negative antibody findings in 12 patients might be due to the lack of blood samples at the later stage of illness. The median seroconversion times for Ab, IgM, and then IgG were days 11, 12, and 4, respectively. The presence of antibodies was <40% among patients within 1 week of onset, and rapidly increased to 100.0% (Ab), 94.3% (IgM), and 79.8% (IgG) by day 15 after onset. In contrast, RNA detectability decreased from 66.7% (58/87) in samples collected before day 7 to 45.5% (25/55) during days 15-39. Combining RNA and antibody detection significantly improved the sensitivity of pathogenic diagnosis for COVID-19 (P < .001), even in the early phase of 1 week from onset (P = .007). Moreover, a higher titer of Ab was independently associated with a worse clinical classification (P = .006). Antibody detection offers vital clinical information during the course of SARS-CoV-2 infection. The findings provide strong empirical support for the routine application of serological testing in the diagnosis and management of COVID-19 patients.


SARS-CoV-2 challenge studies: ethics and risk minimisation.
COVID-19 poses an exceptional threat to global public health and well-being. Recognition of the need to develop effective vaccines at unprecedented speed has led to calls to accelerate research pathways ethically, including by conducting challenge studies (also known as controlled human infection studies (CHIs)) with SARS-CoV-2 (the virus which causes COVID-19). Such research is controversial, with concerns being raised about the social, legal, ethical and clinical implications of infecting healthy volunteers with SARS-CoV-2 for research purposes. Systematic risk evaluations are critical to inform assessments of the ethics of any proposed SARS-CoV-2 CHIs. Such evaluations will necessarily take place within a rapidly changing and at times contested epidemiological landscape, in which differing criteria for the ethical acceptability of research risks have been proposed. This paper critically reviews two such criteria and evaluates whether the use of effective treatment should be a necessary condition for the ethical acceptability of SARS-CoV-2 CHIs, and whether the choice of study sites should be influenced by COVID-19 incidence levels. The paper concludes that ethical evaluations of proposed SARS-CoV-2 CHIs should be informed by rigorous, consultative and holistic approaches to systematic risk assessment.


Discovery of a new gravitational lens system.
A new gravitational lens system, the triple radio source MG2016+112, has been discovered. Five emission lines at a redshift of 3.2733+/-0.0014 have been identified in the spectra of two stellar objects of magnitude 22.5 coincident with radio components 3.4 arc seconds apart. The lines are the narrowest ever observed in objects at such a large redshift. The redshift of a 23rd-magnitude extended optical object coincident with the third radio component has not been determined spectroscopically, but its known optical properties are consistent with those of a giant elliptical galaxy with a redshift of about 0.8.


Expression of membrane anchored cytokines and B7-1 alters tumor microenvironment and induces protective antitumor immunity in a murine breast cancer model.
Many studies have shown that the systemic administration of cytokines or vaccination with cytokine-secreting tumors augments an antitumor immune response that can result in eradication of tumors. However, these approaches are hampered by the risk of systemic toxicity induced by soluble cytokines. In this study, we have evaluated the efficacy of 4TO7, a highly tumorigenic murine mammary tumor cell line, expressing glycosyl phosphatidylinositol (GPI)-anchored form of cytokine molecules alone or in combination with the costimulatory molecule B7-1 as a model for potential cell or membrane-based breast cancer vaccines. We observed that the GPI-anchored cytokines expressed on the surface of tumor cells greatly reduced the overall tumorigenicity of the 4TO7 tumor cells following direct live cell challenge as evidenced by transient tumor growth and complete regression within 30 days post challenge. Tumors co-expressing B7-1 and GPI-IL-12 grew the least and for the shortest duration, suggesting that this combination of immunostimulatory molecules is most potent. Protective immune responses were also observed following secondary tumor challenge. Further, the 4TO7-B7-1/GPI-IL-2 and 4TO7-B7-1/GPI-IL-12 transfectants were capable of inducing regression of a wild-type tumor growing at a distant site in a concomitant tumor challenge model, suggesting the tumor immunity elicited by the transfectants can act systemically and inhibit the tumor growth at a distant site. Additionally, when used as irradiated whole cell vaccines, 4TO7-B7-1/GPI-IL-12 led to a significant inhibition in tumor growth of day 7 established tumors. Lastly, we observed a significant decrease in the prevalence of myeloid-derived suppressor cells and regulatory T-cells in the tumor microenvironment on day 7 post challenge with 4TO7-B7-1/GPI-IL-12 cells, which provides mechanistic insight into antitumor efficacy of the tumor-cell membrane expressed IL-12. These studies have implications in designing membrane-based therapeutic vaccines with GPI-anchored cytokines for breast cancer.


Charge-directed fiber surface modification by molecular assemblies of functional polysaccharides.
Molecular assemblies, namely, polyelectrolyte complexes (PECs) composed of negatively charged xylan-based derivatives and a novel positively charged cellulose derivative (CN(+)), were used for interfacial modification of wood fibers by charge directed self-assembly. The adsorption process was studied using polyelectrolyte titration and elemental analysis. X-ray spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were used as advanced techniques for the characterization of the modified fiber surfaces. The measurements revealed an intense interaction between the pulp fibers and PECs, and provided essential information for a better understanding of the adsorption process. The information gathered on this paper might contribute to the basis for the development of new value added products by the use of underutilized biomass.


Experimental demonstration of the Einstein-Podolsky-Rosen steering game based on the all-versus-nothing proof.
Einstein-Podolsky-Rosen (EPR) steering, a generalization of the original concept of "steering" proposed by Schrödinger, describes the ability of one system to nonlocally affect another system's states through local measurements. Some experimental efforts to test EPR steering in terms of inequalities have been made, which usually require many measurement settings. Analogy to the "all-versus-nothing" (AVN) proof of Bell's theorem without inequalities, testing steerability without inequalities would be more strong and require less resources. Moreover, the practical meaning of steering implies that it should also be possible to store the state information on the side to be steered, a result that has not yet been experimentally demonstrated. Using a recent AVN criterion for two-qubit entangled states, we experimentally implement a practical steering game using quantum memory. Furthermore, we develop a theoretical method to deal with the noise and finite measurement statistics within the AVN framework and apply it to analyze the experimental data. Our results clearly show the facilitation of the AVN criterion for testing steerability and provide a particularly strong perspective for understanding EPR steering.


Neural organization and visual processing in the anterior optic tubercle of the honeybee brain.
The honeybee Apis mellifera represents a valuable model for studying the neural segregation and integration of visual information. Vision in honeybees has been extensively studied at the behavioral level and, to a lesser degree, at the physiological level using intracellular electrophysiological recordings of single neurons. However, our knowledge of visual processing in honeybees is still limited by the lack of functional studies of visual processing at the circuit level. Here we contribute to filling this gap by providing a neuroanatomical and neurophysiological characterization at the circuit level of a practically unstudied visual area of the bee brain, the anterior optic tubercle (AOTu). First, we analyzed the internal organization and neuronal connections of the AOTu. Second, we established a novel protocol for performing optophysiological recordings of visual circuit activity in the honeybee brain and studied the responses of AOTu interneurons during stimulation of distinct eye regions. Our neuroanatomical data show an intricate compartmentalization and connectivity of the AOTu, revealing a dorsoventral segregation of the visual input to the AOTu. Light stimuli presented in different parts of the visual field (dorsal, lateral, or ventral) induce distinct patterns of activation in AOTu output interneurons, retaining to some extent the dorsoventral input segregation revealed by our neuroanatomical data. In particular, activity patterns evoked by dorsal and ventral eye stimulation are clearly segregated into distinct AOTu subunits. Our results therefore suggest an involvement of the AOTu in the processing of dorsoventrally segregated visual information in the honeybee brain.


EPOXI at comet Hartley 2.
Understanding how comets work--what drives their activity--is crucial to the use of comets in studying the early solar system. EPOXI (Extrasolar Planet Observation and Deep Impact Extended Investigation) flew past comet 103P/Hartley 2, one with an unusually small but very active nucleus, taking both images and spectra. Unlike large, relatively inactive nuclei, this nucleus is outgassing primarily because of CO(2), which drags chunks of ice out of the nucleus. It also shows substantial differences in the relative abundance of volatiles from various parts of the nucleus.


Next-generation phage display: integrating and comparing available molecular tools to enable cost-effective high-throughput analysis.
Combinatorial phage display has been used in the last 20 years in the identification of protein-ligands and protein-protein interactions, uncovering relevant molecular recognition events. Rate-limiting steps of combinatorial phage display library selection are (i) the counting of transducing units and (ii) the sequencing of the encoded displayed ligands. Here, we adapted emerging genomic technologies to minimize such challenges. We gained efficiency by applying in tandem real-time PCR for rapid quantification to enable bacteria-free phage display library screening, and added phage DNA next-generation sequencing for large-scale ligand analysis, reporting a fully integrated set of high-throughput quantitative and analytical tools. The approach is far less labor-intensive and allows rigorous quantification; for medical applications, including selections in patients, it also represents an advance for quantitative distribution analysis and ligand identification of hundreds of thousands of targeted particles from patient-derived biopsy or autopsy in a longer timeframe post library administration. Additional advantages over current methods include increased sensitivity, less variability, enhanced linearity, scalability, and accuracy at much lower cost. Sequences obtained by qPhage plus pyrosequencing were similar to a dataset produced from conventional Sanger-sequenced transducing-units (TU), with no biases due to GC content, codon usage, and amino acid or peptide frequency. These tools allow phage display selection and ligand analysis at >1,000-fold faster rate, and reduce costs approximately 250-fold for generating 10(6) ligand sequences. Our analyses demonstrates that whereas this approach correlates with the traditional colony-counting, it is also capable of a much larger sampling, allowing a faster, less expensive, more accurate and consistent analysis of phage enrichment. Overall, qPhage plus pyrosequencing is superior to TU-counting plus Sanger sequencing and is proposed as the method of choice over a broad range of phage display applications in vitro, in cells, and in vivo.


Restoration of Physiologically Responsive Low-Density Lipoprotein Receptor-Mediated Endocytosis in Genetically Deficient Induced Pluripotent Stem Cells.
Acquiring sufficient amounts of high-quality cells remains an impediment to cell-based therapies. Induced pluripotent stem cells (iPSC) may be an unparalleled source, but autologous iPSC likely retain deficiencies requiring correction. We present a strategy for restoring physiological function in genetically deficient iPSC utilizing the low-density lipoprotein receptor (LDLR) deficiency Familial Hypercholesterolemia (FH) as our model. FH fibroblasts were reprogrammed into iPSC using synthetic modified mRNA. FH-iPSC exhibited pluripotency and differentiated toward a hepatic lineage. To restore LDLR endocytosis, FH-iPSC were transfected with a 31 kb plasmid (pEHZ-LDLR-LDLR) containing a wild-type LDLR (FH-iPSC-LDLR) controlled by 10 kb of upstream genomic DNA as well as Epstein-Barr sequences (EBNA1 and oriP) for episomal retention and replication. After six months of selective culture, pEHZ-LDLR-LDLR was recovered from FH-iPSC-LDLR and transfected into Ldlr-deficient CHO-a7 cells, which then exhibited feedback-controlled LDLR-mediated endocytosis. To quantify endocytosis, FH-iPSC ± LDLR were differentiated into mesenchymal cells (MC), pretreated with excess free sterols, Lovastatin, or ethanol (control), and exposed to DiI-LDL. FH-MC-LDLR demonstrated a physiological response, with virtually no DiI-LDL internalization with excess sterols and an ~2-fold increase in DiI-LDL internalization by Lovastatin compared to FH-MC. These findings demonstrate the feasibility of functionalizing genetically deficient iPSC using episomal plasmids to deliver physiologically responsive transgenes.


Serum Vitamin D Is Significantly Inversely Associated with Disease Severity in Caucasian Adults with Obstructive Sleep Apnea Syndrome.
To evaluate vitamin D (25(OH)D) levels in obstructive sleep apnea syndrome (OSAS) and possible relationships to OSAS severity, sleepiness, lung function, nocturnal heart rate (HR), and body composition. We also aimed to compare the 25(OH)D status of a subset of OSAS patients compared to controls matched for important determinants of both OSAS and vitamin D deficiency (VDD). This was a cross-sectional study conducted at an urban, clinical sleep medicine outpatient center. We recruited newly diagnosed, Caucasian adults who had recently undergone nocturnal polysomnography. We compared body mass index (BMI), body composition (bioelectrical impedance analysis), neck circumference, sleepiness (Epworth Sleepiness Scale), lung function, and vitamin D status (serum 25-hydrpoxyvitamin D (25(OH)D) across OSAS severity categories and non-OSAS subjects. Next, using a case-control design, we compared measures of serum 25(OH)D from OSAS cases to non-OSAS controls who were matched for age, gender, skin pigmentation, sleepiness, season, and BMI. 106 adults (77 male; median age = 54.5; median BMI = 34.3 kg/m(2)) resident in Dublin, Ireland (latitude 53°N) were recruited and categorized as non-OSAS or mild/moderate/severe OSAS. 98% of OSAS cases had insufficient 25(OH)D (< 75 nmol/L), including 72% with VDD (< 50 nmol/L). 25(OH)D levels decreased with OSAS severity (P = 0.003). 25(OH)D was inversely correlated with BMI, percent body fat, AHI, and nocturnal HR. Subsequent multivariate regression analysis revealed that 25(OH)D was independently associated with both AHI (P = 0.016) and nocturnal HR (P = 0.0419). Our separate case-control study revealed that 25(OH)D was significantly lower in OSAS cases than matched, non-OSAS subjects (P = 0.001). We observed widespread vitamin D deficiency and insufficiency in a Caucasian, OSAS population. There were significant, independent, inverse relationships between 25(OH)D and AHI as well as nocturnal HR, a known cardiovascular risk factor. Further, 25(OH)D was significantly lower in OSAS cases compared to matched, non-OSAS subjects. We provide evidence that 25(OH)D and OSAS are related, but the role, if any, of replenishment has not been investigated.


Starting a research post: some considerations for ward-based doctors.
Starting a research post after ward-based work can be a leap of faith and, as rewarding as it ultimately is, the initial transition can be a difficult one. This article addresses some of the questions which someone considering research should ask.


Use of mobile phones in ICU--why not ban?
Due to the rapid growth of mobile telecommunications it is predicted that by 2005 there will be 1.6 billion mobile phone users worldwide. The usage of cellphones in Intensive Care Units carries with it a high incidence of interference with a number of medical devices like implantable defibrillators, cardioverters, pacemakers, monitors and other important devices like ventilators. It is in this context that this article will throw a light on complications of cellphones use in the Intensive Care Units and various strategies that can be taken to restrict their use in the Intensive Care Units.


Hypoglycaemic and Antioxidant Effects of Propolis of Chihuahua in a Model of Experimental Diabetes.
Propolis is a bee-collected natural product that has been proven to have various bioactivities. This study tested the effects of a Mexican propolis on streptozotocin-induced diabetes mellitus in a murine model. The results showed that an ethanolic extract of propolis of Chihuahua (EEPCh) significantly inhibited increases in blood glucose and the loss of body weight in diabetic mice. EEPCh increased plasma insulin levels in STZ-diabetic mice, whereas, in untreated diabetic mice, there was no detection of insulin. EEPCh had a high antioxidant capacity (SA50 = 15.75 μg/mL), which was directly related to the concentrations of total phenols (314 mg GAE/g of extract) and flavonoids (6.25 mg QE/g of extract). In addition, increased activities of the enzymes superoxide dismutase, catalase, and glutathione peroxidase were observed in diabetic mice treated with EEPCh. Compounds such as pinocembrin, quercetin, naringin, naringenin, kaempferol, acacetin, luteolin, and chrysin were identified by HPLC-MS analysis. This investigation demonstrated that propolis of Chihuahua possesses hypoglycaemic and antioxidant activities and can alleviate symptoms of diabetes mellitus in mice. These effects may be directly related to the chemical composition of propolis, as most of the compounds identified in propolis are reportedly active in terms of the different parameters evaluated in this work.


The influence of DNA sequence on epigenome-induced pathologies.
Clear cause-and-effect relationships are commonly established between genotype and the inherited risk of acquiring human and plant diseases and aberrant phenotypes. By contrast, few such cause-and-effect relationships are established linking a chromatin structure (that is, the epitype) with the transgenerational risk of acquiring a disease or abnormal phenotype. It is not entirely clear how epitypes are inherited from parent to offspring as populations evolve, even though epigenetics is proposed to be fundamental to evolution and the likelihood of acquiring many diseases. This article explores the hypothesis that, for transgenerationally inherited chromatin structures, "genotype predisposes epitype", and that epitype functions as a modifier of gene expression within the classical central dogma of molecular biology. Evidence for the causal contribution of genotype to inherited epitypes and epigenetic risk comes primarily from two different kinds of studies discussed herein. The first and direct method of research proceeds by the examination of the transgenerational inheritance of epitype and the penetrance of phenotype among genetically related individuals. The second approach identifies epitypes that are duplicated (as DNA sequences are duplicated) and evolutionarily conserved among repeated patterns in the DNA sequence. The body of this article summarizes particularly robust examples of these studies from humans, mice, Arabidopsis, and other organisms. The bulk of the data from both areas of research support the hypothesis that genotypes predispose the likelihood of displaying various epitypes, but for only a few classes of epitype. This analysis suggests that renewed efforts are needed in identifying polymorphic DNA sequences that determine variable nucleosome positioning and DNA methylation as the primary cause of inherited epigenome-induced pathologies. By contrast, there is very little evidence that DNA sequence directly determines the inherited positioning of numerous and diverse post-translational modifications of histone side chains within nucleosomes. We discuss the medical and scientific implications of these observations on future research and on the development of solutions to epigenetically induced disorders.


Oral leukoplakia associated with amalgam restorations.
The World Health Organization has defined oral leukoplakia (OL) as "a white patch or plaque of the oral mucosa that cannot be characterized clinically or pathologically as any other disease". A 21-year-old male with OL presented with a bilateral burning sensation in the buccal mucosa. The patient had amalgam restorations, and an epicutaneous patch test indicated a positive response to amalgam. The amalgam restorations were therefore removed and the cavities were refilled with a composite resin restorative material. During 5 years of follow-up, there was no recurrence of the oral lesions. This case illustrates that amalgam fillings may cause OL lesions. (J Oral Sci 58, 445-448, 2016).


On the interplay of shell structure with low- and high-frequency mechanics of multifunctional magnetic microbubbles.
Polymer-shelled magnetic microbubbles have great potential as hybrid contrast agents for ultrasound and magnetic resonance imaging. In this work, we studied US/MRI contrast agents based on air-filled poly(vinyl alcohol)-shelled microbubbles combined with superparamagnetic iron oxide nanoparticles (SPIONs). The SPIONs are integrated either physically or chemically into the polymeric shell of the microbubbles (MBs). As a result, two different designs of a hybrid contrast agent are obtained. With the physical approach, SPIONs are embedded inside the polymeric shell and with the chemical approach SPIONs are covalently linked to the shell surface. The structural design of hybrid probes is important, because it strongly determines the contrast agent's response in the considered imaging methods. In particular, we were interested how structural differences affect the shell's mechanical properties, which play a key role for the MBs' US imaging performance. Therefore, we thoroughly characterized the MBs' geometric features and investigated low-frequency mechanics by using atomic force microscopy (AFM) and high-frequency mechanics by using acoustic tests. Thus, we were able to quantify the impact of the used SPIONs integration method on the shell's elastic modulus, shear modulus and shear viscosity. In summary, the suggested approach contributes to an improved understanding of structure-property relations in US-active hybrid contrast agents and thus provides the basis for their sustainable development and optimization.


Validity of hip-mounted uniaxial accelerometry with heart-rate monitoring vs. triaxial accelerometry in the assessment of free-living energy expenditure in young children: the IDEFICS Validation Study.
One of the aims of Identification and Prevention of Dietary- and Lifestyle-Induced Health Effects in Children and Infants (IDEFICS) validation study is to validate field measures of physical activity (PA) and energy expenditure (EE) in young children. This study compared the validity of uniaxial accelerometry with heart-rate (HR) monitoring vs. triaxial accelerometry against doubly labeled water (DLW) criterion method for assessment of free-living EE in young children. Forty-nine European children (25 female, 24 male) aged 4-10 yr (mean age: 6.9 ± 1.5 yr) were assessed by uniaxial ActiTrainer with HR, uniaxial 3DNX, and triaxial 3DNX accelerometry. Total energy expenditure (TEE) was estimated using DLW over a 1-wk period. The longitudinal axis of both devices and triaxial 3DNX counts per minute (CPM) were significantly (P < 0.05) associated with physical activity level (PAL; r = 0.51 ActiTrainer, r = 0.49 uniaxial-3DNX, and r = 0.42 triaxial Σ3DNX). Eight-six percent of the variance in TEE could be predicted by a model combining body mass (partial r(2) = 71%; P < 0.05), CPM-ActiTrainer (partial r(2) = 11%; P < 0.05), and difference between HR at moderate and sedentary activities (ModHR - SedHR) (partial r(2) = 4%; P < 0.05). The SE of TEE estimate for ActiTrainer and 3DNX models ranged from 0.44 to 0.74 MJ/days or ∼7-11% of the average TEE. The SE of activity-induced energy expenditure (AEE) model estimates ranged from 0.38 to 0.57 MJ/day or 24-26% of the average AEE. It is concluded that the comparative validity of hip-mounted uniaxial and triaxial accelerometers for assessing PA and EE is similar.


Interleukin 6 is essential for in vivo development of B lineage neoplasms.
Interleukin (IL) 6 has been suggested to be the major cytokine responsible for proliferation of neoplastic plasma cells in both human myeloma and mouse plasmacytoma. Much of the evidence supporting this suggestion is derived from in vitro studies in which the survival or proliferation of some plasma cell tumors has been found to be IL-6 dependent. However, it remains unclear whether this dependency is the consequence of in vivo or in vitro selective pressures that preferentially expand IL-6-responsive tumor cells, or whether it reflects a critical in vivo role for IL-6 in plasma cell neoplasia. To address this question, we have attempted to induce plasma cell tumors in normal mice and in IL-6-deficient mice generated by introduction of a germline-encoded null mutation in the IL-6 gene. The results demonstrate that mice homozygous (+/+) or heterozygous (+/-) for the wild-type IL-6 allele yield the expected incidences of plasma cell tumors. In contrast, mice homozygous for the IL-6-null allele (-/-) are completely resistant to plasma cell tumor development. These studies define the essential role of IL-6 in the development of B lineage tumors in vivo and provide experimental support for continued efforts to modulate this cytokine in the treatment of appropriate human B cell malignancies.


Characterization of the role of cadherin in regulating cell adhesion during sea urchin development.
During development, the modulation of cadherin adhesive function is proposed to control various morphogenetic events including epithelial-mesenchymal conversions and tubulogenesis, although the mechanisms responsible for regulating cadherin activity during these events remain unclear. In order to gain insights into the regulation of cadherin function during morphogenesis, we utilized the sea urchin embryo as a model system to study the regulation of cadherin localization during epithelial-mesenchymal conversion and convergent-extension movements. Polyclonal antibodies raised against the cytoplasmic domain of a cloned sea urchin cadherin recognize three major polypeptides of M(r) 320, 140, and 125 kDa and specifically stain adherens junctions, and to a lesser extent, lateral membrane domains in all epithelial tissues of the embryo. Analysis of embryos during gastrulation demonstrates that changes in cadherin localization are observed in cells undergoing an epithelial-mesenchymal conversion. Ingression of primary mesenchyme cells is accompanied by the rapid loss of junctional cadherin staining and the coincident accumulation of cadherin in intracellular organelles. These data are consistent with the idea that the deadhesion of mesenchymal cells from neighboring epithelial cells involves the regulated endocytosis of cell surface cadherin molecules. Conversely, neither cadherin abundance nor localization is altered in cells of the gut which undergo convergent-extension movements during the formation of the archenteron. This observation indicates that these movements do not require the loss of junctional cadherin molecules. Instead, the necessary balance between adhesion and motility may be achieved by regulating the expression of different subtypes of cadherin molecules or modifying interactions between cadherins and catenins, proteins that bind the cytoplasmic domain of cadherin and are necessary for cadherin adhesive function. To address cadherin function at the molecular level, we used a partial cDNA representing the conserved cytoplasmic domain to identify a novel cadherin molecule in the sea urchin Lytechinus variegatus. The deduced amino acid sequence of LvG-cadherin (for Goliath-cadherin) predicts that it is a transmembrane protein with an apparent relative molecular mass of 303 kDa. The cytoplasmic domain shows significant sequence identity to that of vertebrate classic cadherins. However, the extracellular domain is distinguished from its vertebrate counterparts by both an increased number of cadherin-specific repeats and the presence of four EGF-like repeats proximal to the transmembrane domain. Taken together, these data are consistent with the hypothesis that the sea urchin possesses several cadherins, including a novel member of the cadherin family, and that the dynamic regulation of cadherin localization plays a role in epithelial to mesenchymal conversions during gastrulation.


Structural MRI correlates of amyotrophic lateral sclerosis progression.
Amyotrophic lateral sclerosis (ALS) presents with varying degrees of brain degeneration that can extend beyond the corticospinal tract (CST). Furthermore, the clinical course and progression of ALS varies widely. Brain degeneration detected using structural MRI could reflect disease progression. On study registration, 3-Tesla volumetric MRI and diffusion tensor imaging scans were obtained at baseline in 38 healthy controls and 67 patients with sporadic ALS. Patients had Amyotrophic Lateral Sclerosis Functional Rating Scale-Revised (ALSFRS-R) scores of ≥36 and did not have the chromosome 9, open reading frame 72 repeat expansion. Six months later, changes in ALSFRS-R (ΔALSFRS-R) scores were calculated and patients were grouped into three categories, namely, patients with slow progression with ΔALSFRS-R scores ≤3 (n=19), intermediate progression with ΔALSFRS-R scores =4, 5 and 6 (n=36) and rapid progression with ΔALSFRS-R scores ≥7 (n=12). We analysed voxel-based morphometry and tract-based spatial statistics among these subgroups and controls. In comparison with controls, patients with ALS showed grey matter atrophy and decreased fractional anisotropy beyond the motor cortex and CST, especially in the frontotemporal lobes and basal ganglia. Moreover, the degree of change was highly proportional to ΔALSFRS-R at the 6-month assessment. A more rapid disease progression and poorer functional decline were associated with greater involvement of the extra-motor cortex and basal ganglia, suggesting that the spatial extent of brain involvement can be an indicator of the progression in ALS.


Effect of two self-adhesive cements on marginal adaptation and strength of esthetic ceramic CAD/CAM molar crowns.
This study evaluated the effects of adhesive cements on marginal adaptation and fracture resistance of ceramic molar crowns. Seventy-five extracted maxillary molars were selected. The occlusal morphology of 15 molars (control) was scanned and transferred to the crowns in the test groups by CAD/CAM. Sixty molars received full-coverage crown preparations with 6-degree axial taper, 1.0-mm shoulder, and 2.0-mm occlusal reduction. They were assigned to four groups, and pulpal pressure was simulated. The 15 crowns in each test group were seated with resin-based self-adhesive cements, Rely-X (RX) and Multilink (MS), one multistep bonded adhesive luting composite resin, Variolink (VL), and glass-ionomer cement, Ketac Cem (KC). Test and control molars were subjected to thermal and mechanical fatigue stress (TMS: 12,000 x 5 degrees C to 50 degrees C; 2.4 million x 49 N) for 18 days in a masticator. Marginal adaptation ["continuous margin%" (CM%)] of the crowns was determined by scanning electron microscopy (200x). Finally, molars were occlusally loaded until fracture in a testing machine, and fracture load (N) was recorded. Marginal adaptation and strength data were statistically analyzed. TMS significantly (p < 0.001) reduced CM% in all groups. After TMS, CM% at the cement-dentin interface was significantly (p < 0.001) higher for RX than for all other cements. At the crown-cement interface both self-adhesive cements MS and RX had significantly better CM% than VL (p < 0.05) and KC (p < 0.001). Fracture resistance of natural untreated molars was significantly (p < 0.001) higher than that of experimental crowns. Fracture resistance of RX cemented crowns was significantly (p < 0.05) higher than that of other crowns. Occlusal morphology significantly influenced fracture resistance (p < 0.05). Self-adhesive cement RX offers a valid alternative to multistep resin-based luting composite with respect to marginal adaptation to dentin and fracture resistance. The latter is also influenced by occlusal morphology, necessitating careful monitoring of occlusal contacts.


Relationship between Health Literacy and Sexual Function and Sexual Satisfaction in Infertile Couples Referred to The Royan Institute.
Health science and technology today is a rapidly growing field. Health is a multifaceted concept influenced by several factors, and health literacy is essential to deal properly with the current situation. In this study, the association between health literacy and sexual function and sexual satisfaction were investigated in 2016. This descriptive and correlational study was conducted on 193 couples in the Royan Institute, Tehran. Data collection instruments were three standard questionnaires which included the Test of Functional Health Literacy, the Female Sexual Function Index (FSFI) and the International Index of Erectile Function, and the Iranian version of the Sexual Satisfaction Scale. The data were analyzed using SPSS-v23 software at a significance level of 0.05. Marginal health literacy, 49.7% among men and 44.1% among women, was more common than adequate or inadequate health literacy. Erectile function for the majority of men was appropriate (53.3%), compared to 16.6% who had perfect function and 30.1% for whom function was less than appropriate. The majority of women (57.0%) had sexual dysfunction. One hundred and three (53.3%) men had appropriate sexual function and 57% of women had normal sexual function. The greater proportion of men (50.8%) and women (46.1%) had good, rather than very good or less than good, sexual satisfaction. The results of chi-square tests indicated that greater health literacy was associated with higher levels of sexual function and sexual satisfaction among men and women. However, application of the Cramer's V test indicates that the strength of these associations is moderate to weak. Health literacy was marginal among most couples and its adverse impacts on sexual function and sexual satisfaction were confirmed. Accordingly, it is recommended that plans be developed to promote health literacy among infertile couples.


Zoonotic babesiosis: overview of the disease and novel aspects of pathogen identity.
Babesiosis is a zoonosis caused by tick-transmitted intraerythrocytic protozoa of the Phylum Apicomplexa. The disease mostly occurs in the USA, but cases have also been reported in several European countries, in Egypt, India, Japan, Korea, Taiwan, and South Africa. The main pathological event is lysis of erythrocytes resulting in haemolytic anaemia, which in severe cases may lead to organ failure and death, particularly in immunocompromised patients. The 2 groups of parasites involved, Babesia microti-like and Babesia sensu stricto (s.s.) species, differ in their life cycle characteristics and susceptibility to antibabesial drugs. Molecular taxonomy is now making a major contribution to the identification of novel pathogens within both groups. Effective treatment of severe cases was initially hampered by the lack of specific antibabesial drugs for human use, but increased use of supportive measures and of the recently developed antimalarial, atovaquone, particularly in combination with azithromycin, has improved the prospects for management of acute disease especially when caused by Babesia s.s. species. Prevention should be based primarily on increasing the awareness of physicians and the public to the risks, but infection from blood transfusions is particularly difficult to prevent. Expanding deer populations, resulting in wider distribution and greater abundance of ticks, heightened medical awareness, and growing numbers of immunocompromised patients are likely to result in a continuing rise of reported cases.


Deletion of AU-rich elements within the Bcl2 3'UTR reduces protein expression and B cell survival in vivo.
Post-transcriptional mRNA regulation by RNA binding proteins (RBPs) associated with AU-rich elements (AREs) present in the 3' untranslated region (3'UTR) of specific mRNAs modulates transcript stability and translation in eukaryotic cells. Here we have functionally characterised the importance of the AREs present within the Bcl2 3'UTR in order to maintain Bcl2 expression. Gene targeting deletion of 300 nucleotides of the Bcl2 3'UTR rich in AREs diminishes Bcl2 mRNA stability and protein levels in primary B cells, decreasing cell lifespan. Generation of chimeric mice indicates that Bcl2-ARE∆/∆ B cells have an intrinsic competitive disadvantage compared to wild type cells. Biochemical assays and predictions using a bioinformatics approach show that several RBPs bind to the Bcl2 AREs, including AUF1 and HuR proteins. Altogether, association of RBPs to Bcl2 AREs contributes to Bcl2 protein expression by stabilizing Bcl2 mRNA and promotes B cell maintenance.


How accurate and robust are the phylogenetic estimates of Austronesian language relationships?
We recently used computational phylogenetic methods on lexical data to test between two scenarios for the peopling of the Pacific. Our analyses of lexical data supported a pulse-pause scenario of Pacific settlement in which the Austronesian speakers originated in Taiwan around 5,200 years ago and rapidly spread through the Pacific in a series of expansion pulses and settlement pauses. We claimed that there was high congruence between traditional language subgroups and those observed in the language phylogenies, and that the estimated age of the Austronesian expansion at 5,200 years ago was consistent with the archaeological evidence. However, the congruence between the language phylogenies and the evidence from historical linguistics was not quantitatively assessed using tree comparison metrics. The robustness of the divergence time estimates to different calibration points was also not investigated exhaustively. Here we address these limitations by using a systematic tree comparison metric to calculate the similarity between the Bayesian phylogenetic trees and the subgroups proposed by historical linguistics, and by re-estimating the age of the Austronesian expansion using only the most robust calibrations. The results show that the Austronesian language phylogenies are highly congruent with the traditional subgroupings, and the date estimates are robust even when calculated using a restricted set of historical calibrations.


Innate lymphoid cells--a proposal for uniform nomenclature.
Innate lymphoid cells (ILCs) are a family of developmentally related cells that are involved in immunity and in tissue development and remodelling. Recent research has identified several distinct members of this family. Confusingly, many different names have been used to characterize these newly identified ILC subsets. Here, we propose that ILCs should be categorized into three groups based on the cytokines that they can produce and the transcription factors that regulate their development and function.


Fried-food consumption and risk of cardiovascular disease and all-cause mortality: a meta-analysis of observational studies.
We performed a meta-analysis, including dose-response analysis, to quantitatively determine the association of fried-food consumption and risk of cardiovascular disease and all-cause mortality in the general adult population. We searched PubMed, EMBASE and Web of Science for all articles before 11 April 2020. Random-effects models were used to estimate the summary relative risks (RRs) and 95% CIs. In comparing the highest with lowest fried-food intake, summary RRs (95% CIs) were 1.28 (1.15 to 1.43; n=17, I2=82.0%) for major cardiovascular events (prospective: 1.24 (1.12 to 1.38), n=13, I2=75.7%; case-control: 1.91 (1.15 to 3.17), n=4, I2=92.1%); 1.22 (1.07 to 1.40; n=11, I2=77.9%) for coronary heart disease (prospective: 1.16 (1.05 to 1.29), n=8, I2=44.6%; case-control: 1.91 (1.05 to 3.47), n=3, I2=93.9%); 1.37 (0.97 to 1.94; n=4, I2=80.7%) for stroke (cohort: 1.21 (0.87 to 1.69), n=3, I2=77.3%; case-control: 2.01 (1.27 to 3.19), n=1); 1.37 (1.07 to 1.75; n=4, I2=80.0%) for heart failure; 1.02 (0.93 to 1.14; n=3, I2=27.3%) for cardiovascular mortality; and 1.03 (95% CI 0.96 to 1.12; n=6, I2=38.0%) for all-cause mortality. The association was linear for major cardiovascular events, coronary heart disease and heart failure. Fried-food consumption may increase the risk of cardiovascular disease and presents a linear dose-response relation. However, the high heterogeneity and potential recall and misclassification biases for fried-food consumption from the original studies should be considered.


Abstractions for DNA circuit design.
DNA strand displacement techniques have been used to implement a broad range of information processing devices, from logic gates, to chemical reaction networks, to architectures for universal computation. Strand displacement techniques enable computational devices to be implemented in DNA without the need for additional components, allowing computation to be programmed solely in terms of nucleotide sequences. A major challenge in the design of strand displacement devices has been to enable rapid analysis of high-level designs while also supporting detailed simulations that include known forms of interference. Another challenge has been to design devices capable of sustaining precise reaction kinetics over long periods, without relying on complex experimental equipment to continually replenish depleted species over time. In this paper, we present a programming language for designing DNA strand displacement devices, which supports progressively increasing levels of molecular detail. The language allows device designs to be programmed using a common syntax and then analysed at varying levels of detail, with or without interference, without needing to modify the program. This allows a trade-off to be made between the level of molecular detail and the computational cost of analysis. We use the language to design a buffered architecture for DNA devices, capable of maintaining precise reaction kinetics for a potentially unbounded period. We test the effectiveness of buffered gates to support long-running computation by designing a DNA strand displacement system capable of sustained oscillations.


Group psychotherapies for depression in persons with HIV: A systematic review.
Studies investigating effectiveness of group psychotherapy intervention in depression in persons with HIV have showed varying results with differing effect sizes. A systematic review of randomized controlled trials of group psychotherapy in depression in persons with HIV has been conducted to present the best available evidence in relation to its effect on depressive symptomatology. Electronic databases were searched to identify randomized controlled trials. Selected studies were quality assessed and data extracted by two reviewers. If feasible, it was planned to conduct a meta-analysis to obtain a pooled effect size of group psychotherapeutic interventions on depressive symptoms. Odds ratio for drop out from group was calculated. The studies were assessed for their quality using the Quality Rating Scale and other parameters for quality assessment set out by COCHRANE. The quality of reporting of the trials was compared against the Consolidated Standards of Reporting Trials (CONSORT) checklist for non-pharmacological studies (CONSORT-NPT). Four studies met the full inclusion criteria for systematic review. The trials included in the review examined group interventions based on the Cognitive behavioral therapy model against other therapeutic interventions or waiting list controls. In all four studies, group psychotherapy was an effective intervention for reducing depressive symptoms in persons with HIV in comparison to waiting list controls. The reported benefits from the group psychotherapy in comparison to active controls were less impressive. There were no statistically significant differences in drop outs at post treatments across group psychotherapy, wait list control, and other active interventions. The methodological quality of the studies varied. The quality of reporting of the studies was sub-optimal. The results of this systematic review support that group psychological interventions for depression in persons with HIV have a significant effect on depressive symptomatology. This review also indicates that group cognitive behavioral therapies are an acceptable psychological intervention for persons with HIV and comorbid depression.


Nonparametric dynamical model of cardiorespiratory responses at the onset and offset of treadmill exercises.
This paper applies a nonparametric modelling method with kernel-based regularization to estimate the carbon dioxide production during jogging exercises. The kernel selection and regularization strategies have been discussed; several commonly used kernels are compared regarding the goodness-of-fit, sensitivity, and stability. Based on that, the most appropriate kernel is then selected for the construction of the regularization term. Both the onset and offset of the jogging exercises are investigated. We compare the identified nonparametric models, which include both impulse response models and step response models for the two periods, as well as the relationship between oxygen consumption and carbon dioxide production. The result statistically indicates that the steady-state gain of the carbon dioxide production in the onset of exercise is bigger than that in the offset while the response time of both onset and offset are similar. Compared with oxygen consumption, the response speed of carbon dioxide production is slightly slower in both onset and offset period while its steady-state gains are similar for both periods. The effectiveness of the kernel-based method for the dynamic modelling of cardiorespiratory response to exercise is also well demonstrated. Graphical Abstract Comparison between VO2 and VCO2 during onset and offset of exercise.


No Protective Effect of Constitutive Activation of AMPK in Endothelial Cells on Vascular Function in Aged Obese Mice but Augmented α1-Adrenergic Contractions in Renal Arteries Reversible by Weight Loss.
Aging, obesity, and diabetes favor vascular dysfunction. Endothelial activation of adenosine monophosphate-activated protein kinase (AMPK) has protective effects in diabetes. Mice with constitutive endothelial activation of AMPK (CA-AMPK) were given a high fat diet to induce obesity or kept on standard chow as lean controls for up to 2 years. A subset of obese animals was changed to standard chow after 30 weeks of high fat feeding. En-dothelium-dependent and endothelium-independent responses were examined by isometric tension recording. Endothelium-dependent nitric oxide (NO)- and apamin plus charybdotoxin-sensitive relaxations were preserved and similar between aortic or renal arterial preparations of lean and obese CA-AMPK mice and their wild-type littermates. Despite comparable release of vasoconstrictor prostanoids, cyclooxygenase-dependent contractions were enhanced during NO synthase inhibition in carotid arterial rings of obese CA-AMPK mice. Contractions to the α1-adrenoceptor agonist phenylephrine were augmented in renal arteries of obese animals, a genotype-independent phenomenon reversible by weight loss. These data indicate a higher α1-adrenergic reactivity in renal arteries of aged mice with obesity. The current results highlight the potential of weight loss to alleviate vascular dysfunction. However, endothelial activation of the AMPK pathway in obesity may not be sufficient to prevent vascular dysfunction without lifestyle changes.


Involvement of phytosulfokine in the attenuation of stress response during the transdifferentiation of zinnia mesophyll cells into tracheary elements.
Phytosulfokine (PSK) is a sulfated peptide hormone required for the proliferation and differentiation of plant cells. Here, we characterize the physiological roles of PSK in transdifferentiation of isolated mesophyll cells of zinnia (Zinnia elegans 'Canary Bird') into tracheary elements (TEs). Transcripts for a zinnia PSK precursor gene, ZePSK1, show two peaks of expression during TE differentiation; the first accumulation is transiently induced in response to wounding at 24 h of culture, and the second accumulation is induced in the final stage of TE differentiation and is dependent on endogenous brassinosteroids. Chlorate, a potent inhibitor of peptide sulfation, is successfully applied as an inhibitor of PSK action. Chlorate significantly suppresses TE differentiation. The chlorate-induced suppression of TE differentiation is overcome by exogenously applied PSK. In the presence of chlorate, expression of stress-related genes for proteinase inhibitors and a pathogenesis-related protein is enhanced and changed from a transient to a continuous pattern. On the contrary, administration of PSK significantly reduces the accumulation of transcripts for the stress-related genes. Even in the absence of auxin and cytokinin, addition of PSK suppresses stress-related gene expression. Microarray analysis reveals 66 genes down-regulated and 42 genes up-regulated in the presence of PSK. The large majority of down-regulated genes show significant similarity to various families of stress-related proteins, including chitinases, phenylpropanoid biosynthesis enzymes, 1-aminocyclopropane-1-carboxylic acid synthase, and receptor-like protein kinases. These results suggest the involvement of PSK in the attenuation of stress response and healing of wound-activated cells during the early stage of TE differentiation.


Multi-stress resistance in Lactococcus lactis is actually escape from purine-induced stress sensitivity.
Multi-stress resistance is a widely documented and fascinating phenotype of lactococci where single mutations, preferentially in genes involved in nucleotide metabolism and phosphate uptake, result in elevated tolerance to multiple stresses simultaneously. In this report, we have analysed the metabolic basis behind this multi-stress-resistance phenotype in Lactococcus lactis subsp. cremoris MG1363 using acid stress as a model of multi-stress resistance. Surprisingly, we found that L. lactis MG1363 is fully resistant to pH 3.0 in the chemically defined SA medium, contrary to its sensitivity in the rich and complex M17 medium. When salvage of purines and subsequent conversion to GTP was permitted in various genetic backgrounds of L. lactis MG1363, the cells became sensitive to acid stress, indicating that an excess of guanine nucleotides induces stress sensitivity. The addition of phosphate to the acid-stress medium increased the stress sensitivity of L. lactis MG1363. It is also shown that high intracellular guanine nucleotide pools confer increased sensitivity to high temperatures, thus showing that it is indeed a multi-stress phenotype. Our analysis suggests that an increased level of guanine nucleotides is formed as a result of an improved conversion of guanosine in the salvage pathway. Based upon our findings, we suggest that L. lactis MG1363 is naturally multi-stress resistant in habitats devoid of any purine source. However, any exogenous purine that results in increased guanine nucleotide pools renders the bacterium sensitive to environmental stresses.


A prospective study of diet quality and mental health in adolescents.
A number of cross-sectional and prospective studies have now been published demonstrating inverse relationships between diet quality and the common mental disorders in adults. However, there are no existing prospective studies of this association in adolescents, the onset period of most disorders, limiting inferences regarding possible causal relationships. In this study, 3040 Australian adolescents, aged 11-18 years at baseline, were measured in 2005-6 and 2007-8. Information on diet and mental health was collected by self-report and anthropometric data by trained researchers. There were cross-sectional, dose response relationships identified between measures of both healthy (positive) and unhealthy (inverse) diets and scores on the emotional subscale of the Pediatric Quality of Life Inventory (PedsQL), where higher scores mean better mental health, before and after adjustments for age, gender, socio-economic status, dieting behaviours, body mass index and physical activity. Higher healthy diet scores at baseline also predicted higher PedsQL scores at follow-up, while higher unhealthy diet scores at baseline predicted lower PedsQL scores at follow-up. Improvements in diet quality were mirrored by improvements in mental health over the follow-up period, while deteriorating diet quality was associated with poorer psychological functioning. Finally, results did not support the reverse causality hypothesis. This study highlights the importance of diet in adolescence and its potential role in modifying mental health over the life course. Given that the majority of common mental health problems first manifest in adolescence, intervention studies are now required to test the effectiveness of preventing the common mental disorders through dietary modification.


Treadmill training for patients with Parkinson's disease.
Treadmill training is used in rehabilitation and is described as improving gait parameters of patients with Parkinson's disease. To assess the effectiveness of treadmill training in improving the gait of patients with Parkinson's disease and the acceptability and safety of this type of therapy. We searched the Cochrane Movement Disorders Group Specialised Register (see Review Group details for more information) (last searched September 2014), Cochrane Central Register of Controlled Trials (The Cochrane Library 2014, Issue 10), MEDLINE (1950 to September 2014), and EMBASE (1980 to September 2014). We also handsearched relevant conference proceedings, searched trials and research registers, and checked reference lists (last searched September 2014). We contacted trialists, experts and researchers in the field and manufacturers of commercial devices. We included randomised controlled trials comparing treadmill training with no treadmill training in patients with Parkinson's disease. Two review authors independently selected trials for inclusion, assessed trial quality and extracted data. We contacted the trialists for additional information. We analysed the results as mean differences (MDs) for continuous variables and relative risk differences (RD) for dichotomous variables. We included 18 trials (6 3 3 participants) in this update of this review. Treadmill training improved gait speed (MD = 0.09 m/s; 95% confidence interval (CI) 0.03 to 0.14; P = 0.001; I(2) = 24%; m oderate quality of evidence), stride length (MD = 0.05 metres; 95% CI 0.01 to 0.09; P = 0.01; I(2) = 0%; l ow quality of e vidence), but walking distance (MD = 48.9 metres; 95% CI -1.32 to 99.14; P = 0.06; I(2) = 91%; very low quality of evidence) and cadence did not improve (MD = 2.16 steps/minute; 95% CI -0.13 to 4.46; P = 0.07; I(2) = 28%; low quality of evidence) at the end of study. Treadmill training did not increase the risk of patients dropping out from intervention (RD = -0.02; 95% CI -0.06 to 0.02; P = 0.32; I(2) = 1 3%; m oderate quality of evidence). Adverse events were not reported in included studies. This update of our systematic review provides evidence from e ighteen trials with moderate to l ow risk of bias that the use of treadmill training in patients with PD may improve clinically relevant gait parameters such as gait speed and stride length (m oderate and low quality of evidence, respectively) . This apparent benefit for patients is, however, not supported by all secondary variables (e.g. cadence and walking distance ). Comparing physiotherapy and treadmill training against other alternatives in the treatment of gait hypokinesia such as physiotherapy without treadmill training this type of therapy seems to be more beneficial in practice without increased risk. The gain seems small to moderate clinically relevant . However, the results must be interpreted with caution because it is not known how long these improvements may last and some s tudies used no intervention in the control group and underlie some risk of bias . Additionally the results were heterogenous and we found variations between the trials in patient characteristics, the duration and amount of training, and types of treadmill training applied.


Historical demography of Mullerian mimicry in the neotropical Heliconius butterflies.
We compare the historical demographies of two Müllerian comimetic butterfly species: Heliconius erato and Heliconius melpomene. These species show an extensive parallel geographic divergence in their aposematic wing phenotypes. Recent studies suggest that this coincident mosaic results from simultaneous demographic processes shaped by extrinsic forces over Pleistocene climate fluctuations. However, DNA sequence variation at two rapidly evolving unlinked nuclear loci, Mannose phosphate isomerase (Mpi) and Triose phosphate isomerase (Tpi), show that the comimetic species have quite different quaternary demographies. In H. erato, despite ongoing lineage sorting across the Andes, nuclear genealogical estimates showed little geographical structure, suggesting high historical gene flow. Coalescent-based demographic analysis revealed population growth since the Pliocene period. Although these patterns suggest vicariant population subdivision associated with the Andean orogeny, they are not consistent with hypotheses of Pleistocene population fragmentation facilitating allopatric wing phenotype radiation in H. erato. In contrast, nuclear genetic diversity, theta, in H. melpomene was reduced relative to its comimic and revealed three phylogeographical clades. The pattern of coalescent events within regional clades was most consistent with population growth in relatively isolated populations after a recent period of restricted population size. These different demographic histories suggest that the wing-pattern radiations were not coincident in the two species. Instead, larger effective population size (N(e)) in H. erato, together with profound population change in H. melpomene, supports an earlier hypothesis that H. erato diversified first as the model species of this remarkable mimetic association.


A quantitative analysis of swimming pool recirculation system efficiency.
The primary objective of this study was to conduct a quantitative analysis of the hydraulic efficiency of a 1:25 bench-scale swimming pool and to determine whether the recirculation efficiency could be increased by modifying parameters such as turnover rate, inlet/out configuration, and extent of mixing within the pool. Salt tracer studies were conducted using KCl to determine the residence time distribution and describe the hydraulic characteristics of the pool. The results indicated that the removal of the tracer always followed an exponential decay curve, i.e. 63, 86, 95% for the first, second, and third turnover periods, respectively. In the majority of experiments, the exponential decay rate matched the inverse of the theoretical hydraulic detention time of the system. The results showed that none of the investigated parameters had any significant impact on the tracer removal efficiency. Increasing removal efficiencies of current treatment technologies such as sand and cartridge filters from approximately 25-90% would provide significant improvements in the rate of removal of Cryptosporidium-sized particles. Improving the treatment efficiency beyond 90% would have little additional impact, but further improvements could be achieved by decreasing the system turnover rate.


Phylogenetic findings suggest possible new habitat and routes of infection of human eumyctoma.
Eumycetoma is a traumatic fungal infection in tropical and subtropical areas that may lead to severe disability. Madurella mycetomatis is one of the prevalent etiologic agents in arid Northeastern Africa. The source of infection has not been clarified. Subcutaneous inoculation from plant thorns has been hypothesized, but attempts to detect the fungus in relevant material have remained unsuccessful. The present study aims to find clues to reveal the natural habitat of Madurella species using a phylogenetic approach, i.e. by comparison of neighboring taxa with known ecology. Four species of Madurella were included in a large data set of species of Chaetomium, Chaetomidium, Thielavia, and Papulaspora (n = 128) using sequences of the universal fungal barcode gene rDNA ITS and the partial LSU gene sequence. Our study demonstrates that Madurella species are nested within the Chaetomiaceae, a family of fungi that mainly inhabit animal dung, enriched soil, and indoor environments. We hypothesize that cattle dung, ubiquitously present in rural East Africa, plays a significant role in the ecology of Madurella. If cow dung is an essential factor in inoculation by Madurella, preventative measures may involve the use of appropriate footwear in addition to restructuring of villages to reduce the frequency of contact with etiologic agents of mycetoma. On the other hand, the Chaetomiaceae possess a hidden clinical potential which needs to be explored.


Surveillance for elevated blood lead levels among children--United States, 1997-2001.
Lead is neurotoxic and particularly harmful to the developing nervous systems of fetuses and young children. Extremely high blood lead levels (BLLs) (i.e.,> or =70 microg/dL) can cause severe neurologic problems (e.g., seizure, coma, and death). However, no threshold has been determined regarding lead's harmful effects on children's learning and behavior. In 1990, the U.S. Department of Health and Human Services established a national goal to eliminate BLLs >25 microg/dL by 2000; a new goal targets elimination of BLLs > or =10 microg/dL in children aged <6 years by 2010. Information regarding children's BLLs comes from 1) National Health and Nutrition Examination Surveys (NHANES) conducted during 1976-1980, 1988-1991, 1991-1994, and 1999-2000; and 2) state child blood lead surveillance data for test results collected during 1997-2001. CDC tracks children's BLLs in the United States by using both NHANES and state and local surveillance data. NHANES reports data regarding children aged 1-5 years; state and local surveillance systems report data regarding children aged <72 months. Because lead exposure in children varies among populations and communities, both surveys are needed to determine the burden of elevated BLLs among young children throughout the United States. NHANES uses highly standardized data-collection procedures and probability samples to gather information regarding the health and nutritional status of the civilian, noninstitutionalized U.S. population. State and local childhood lead surveillance systems are based on reports of blood lead tests from laboratories. State and local programs submit data to CDC annually. In this report, confirmed elevated BLLs are defined as one venous blood specimen > or =10 microg/dL or two capillary blood specimens > or =10 microg/dL drawn within 12 weeks of each other. The NHANES 1999-2000 survey estimated that 434,000 children (95% confidence interval = 189,000-846,000) or 2.2% of children aged 1-5 years had BLLs > or =10 microg/dL. For 2001, a total of 44 states, the District of Columbia (DC), and New York City (NYC) submitted child blood lead surveillance data to CDC. These jurisdictions represent 95% of the U.S. population of children aged <72 months and 97% of the nation's pre-1950 housing. The number of children tested and reported to CDC increased from 1,703,356 in 1997 (37 states, DC, and NYC reporting), to 2,422,298 in 2001 (44 states, DC, and NYC reporting). During that time, the number of children reported with confirmed elevated BLLs > or =10 microg/dL steadily decreased from 130,512 in 1997 to 74,887 in 2001. In 2000, the year targeted for national elimination of BLLs >25 microg/dL, a total of 8,723 children had BLLs > or =25 microg/dL. Both national surveys and state surveillance data indicate children's BLLs continue to decline throughout the United States. However, thousands of children continue to be identified with elevated BLLs. The 2000 goal of eliminating BLLs >25 microg/dL was not met. Attaining the 2010 goal of eliminating BLLs > or =10 microg/dL will require intensified efforts to target areas at highest risk, evaluate preventive measures, and improve the quality of surveillance data. States will continue to use surveillance data to 1) promote legislation supporting lead poisoning prevention activities, 2) obtain funding, 3) identify risk groups, 4) target and evaluate prevention activities, and 5) monitor and describe progress toward elimination of BLLs > or =10 microg/dL. CDC will work with state and local programs to improve tracking systems and the collection, timeliness, and quality of surveillance data.


Altered porphyrin excretion and histopathology of greylag geese (Anser anser) exposed to soil contaminated with lead and arsenic in the Guadalquivir Marshes, southwestern Spain.
Greylag geese (Anser anser) in the Guadalquivir Marshes (southwestern Spain) can be exposed to sources of inorganic pollution such as heavy metals and arsenic from mining activities or Pb shot used for hunting. We have sampled 270 fecal excreta in different areas of the marshes in 2001 to 2002 to evaluate the exposure to Pb, Zn, Cu, Mn, and As and to determine its relationship with soil ingestion and with the excretion of porphyrins and biliverdin as biomarkers. These effects and the histopathology of liver, kidney, and pancreas were also studied in 50 geese shot in 2002 to 2004. None of the geese had ingested Pb shot in the gizzard. This contrasts with earlier samplings before the ban of Pb shot for waterfowl hunting in 2001 and the removal of Pb shot in points of the Doñana National Park (Spain) in 1999 to 2000. The highest exposure through direct soil ingestion to Pb and other studied elements was observed in samples from Entremuros, the area of the Doñana Natural Park affected by the Aznalcóllar mine spill in 1998. Birds from Entremuros also more frequently showed mononuclear infiltrates in liver and kidney than birds from the unaffected areas, although other more specific lesions of Pb or Zn poisoning were not observed. The excretion of coproporphyrins, especially of the isomer I, was positively related to the fecal As concentration, and the ratio of coproporphyrin III/I was positively related to fecal Pb concentration. Biliary protoporphyrin IX concentration was also slightly related to hepatic Pb concentration. This study reflects biological effects on terrestrial animals by the mining pollution in Doñana that can be monitored with the simple noninvasive sampling of feces.


Arsenic stress after the Proterozoic glaciations.
Protection against arsenic damage in organisms positioned deep in the tree of life points to early evolutionary sensitization. Here, marine sedimentary records reveal a Proterozoic arsenic concentration patterned to glacial-interglacial ages. The low glacial and high interglacial sedimentary arsenic concentrations, suggest deteriorating habitable marine conditions may have coincided with atmospheric oxygen decline after ~2.1 billion years ago. A similar intensification of near continental margin sedimentary arsenic levels after the Cryogenian glaciations is also associated with amplified continental weathering. However, interpreted atmospheric oxygen increase at this time, suggests that the marine biosphere had widely adapted to the reorganization of global marine elemental cycles by glaciations. Such a glacially induced biogeochemical bridge would have produced physiologically robust communities that enabled increased oxygenation of the ocean-atmosphere system and the radiation of the complex Ediacaran-Cambrian life.