The shape of the global causes of death.
World maps can provide an instant visual overview of the distribution of diseases and deaths. There is a particular geography to each type of death: in some places many thousands of deaths are caused by a particular condition, whilst other equally populous areas have few to no deaths from the same cause. Physicians and other health professionals often specialise in the specifics of causes, symptoms and effects. For some practitioners gaining a worldview of disease burden complements smaller scale medical knowledge of where and how people are affected by each condition. Maps can make health related information much more accessible to planners and the general public than can tables, text, or even graphs. Ten cartograms based on World Health Organisation Burden of Disease data are introduced here; alongside seven based on data from other sources. The Burden of Disease cartograms are the latest in a much larger collection of social, economic and health world maps.


Structure of CrmE, a virus-encoded tumour necrosis factor receptor.
Vaccinia virus (VACV), the smallpox vaccine, encodes many proteins that subvert the host immune response. One of these, cytokine response modifier E (CrmE), is secreted by infected cells and protects these cells from apoptotic challenge by tumour necrosis factor alpha (TNFalpha). We have expressed recombinant CrmE from VACV strain Lister in Escherichia coli, shown that the purified protein is monomeric in solution and competent to bind TNFalpha, and solved the structure to 2.0 A resolution. This is the first structure of a virus-encoded tumour necrosis factor receptor (TNFR). CrmE shares significant sequence similarity with mammalian type 2 TNF receptors (TNFSFR1B, p75; TNFR type 2). The structure confirms that CrmE adopts the canonical TNFR fold but only one of the two "ligand-binding" loops of TNFRSF1A is conserved in CrmE, suggesting a mechanism for the higher affinity of poxvirus TNFRs for TNFalpha over lymphotoxin-alpha. The roles of dimerisation and pre-ligand-assembly domains (PLADs) in poxvirus and mammalian TNFR activity are discussed.


Current comorbidity of psychiatric disorders among DSM-IV major depressive disorder patients in psychiatric care in the Vantaa Depression Study.
While numerous studies have documented the high comorbidity of major depressive disorder (MDD) with individual mental disorders, no published study has reported overall current comorbidity with all Axis I and II disorders among psychiatric patients with MDD, nor systematically investigated variations in current comorbidity by sociodemographic factors, inpatient versus outpatient status, and number of lifetime depressive episodes. Psychiatric outpatients and inpatients in Vantaa, Finland, were prospectively screened for an episode of DSM-IV MDD, and 269 patients with a new episode of MDD were enrolled in the Vantaa Depression MDD Cohort Study. Axis I and II comorbidity was assessed via semistructured Schedules for Clinical Assessment in Neuropsychiatry, version 2.0, and Structured Clinical Interview for DSM-II-R personality disorders interviews. The great majority (79%) of patients with MDD suffered from 1 or more current comorbid mental disorders, including anxiety disorder (57%), alcohol use disorder (25%), and personality disorder (44%). Several anxiety disorders were associated with specific Axis II clusters, and panic disorder with agoraphobia was associated with inpatient status. The prevalence of personality disorders varied with inpatient versus outpatient status, number of lifetime depressive episodes, and type of residential area, and the prevalence of substance use disorders varied with gender and inpatient versus outpatient status. Most psychiatric patients with MDD have at least 1 current comorbid disorder. Comorbid disorders are associated not only with other comorbid disorders, but also with sociodemographic factors, inpatient versus outpatient status, and lifetime number of depressive episodes. The influence of these variations on current comorbidity patterns among MDD patients needs to be taken account of in treatment facilities.


Analysis of EGFR, HER2, and TOP2A gene status and chromosomal polysomy in gastric adenocarcinoma from Chinese patients.
The EGFR and HER2 genes are located on chromosomes 7 and 17, respectively. They are therapeutic targets in some tumors. The TOP2A gene, which is located near HER2 on chromosome 17, is the target of many chemotherapeutic agents, and co-amplification of HER2 and TOP2A has been described in several tumor types. Herein, we investigated the gene status of EGFR, HER2, and TOP2A in Chinese gastric carcinoma patients. We determined the rate of polysomy for chromosomes 7 and 17, and we attempted to clarify the relationship between EGFR, HER2, and TOP2A gene copy number and increased expression of their encoded proteins. Furthermore, we tried to address the relationship between alterations in EGFR, HER2, and TOP2A and chromosome polysomy. One hundred cases of formalin fixed and paraffin embedded tumor tissues from Chinese gastric carcinoma patients were investigated by immunohistochemistry and fluorescence in situ hybridization (FISH) methods. Forty-two percent of the cases showed EGFR overexpression; 16% showed EGFR FISH positive; 6% showed HER2 overexpression; and 11% showed HER2 gene amplification, including all six HER2 overexpression cases. TOP2A nuclear staining (nuclear index, NI) was determined in all 100 tumors: NI values ranged from 0.5-90%. Three percent of the tumors showed TOP2A gene amplification, which were all accompanied by HER2 gene amplification. Nineteen percent of the tumors showed chromosome 7 polysomy, and 16% showed chromosome 17 polysomy. Chromosome 7 polysomy correlated significantly with EGFR FISH-positivity, but was not associated with EGFR overexpression. HER2 overexpression associated significantly with HER2 gene amplification. TOP2A gene amplification was significantly associated with HER2 gene amplification. No relationship was found between alterations in the EGFR, HER2, and TOP2A genes and clinicopathologic variables of gastric carcinoma. The data from our study suggest that chromosome 7 polysomy may be responsible for increased EGFR gene copy number in gastric carcinomas, and that HER2 gene amplification may be the major reason for HER2 protein overexpression. A combined investigation of the gene status of EGFR, HER2, and TOP2A should facilitate the identification of a target therapeutic regimen for gastric carcinoma patients.


Using gender analysis to build voluntary counselling and testing responses in Kenya.
The rapid expansion of voluntary counselling and testing (VCT) for HIV in sub-Saharan Africa has led to concerns over the quality and equity of the services. Kenya has seen an unprecedented scale-up of VCT, and valuable lessons have been learnt at national as well as at district and community levels. We combined quantitative and qualitative research methodology and showed how the results of gender analysis can be used to develop equity in VCT scale-up. A gender-disaggregated analysis of VCT client data was conducted for the first 8 months of 2003. These quantitative data revealed that despite an increased vulnerability to HIV, women are underrepresented in VCT sites in all settings in Kenya. Our data also showed that women were also less likely to use condoms or to take home condoms after a VCT visit than their male counterparts. Further exploration through in-depth qualitative work with women and men allowed a better understanding of the reasons behind gender differences in Kenyan VCT sites and helped to develop strategies to address gender inequity. We conclude that there is an ongoing need to mainstream gender in monitoring and evaluation strategies to ensure services meet the needs and priorities of all groups.


Delayed Onset of Symptoms Through Feedback Interference in Chronic Cancers.
In many cancers, such as Chronic Myelogenous Leukemia (CML), pancreatic, and colorectal cancer, long delays exist between the initiation of the disease and the onset of debilitating symptoms. The early stages of these diseases present manageable symptoms and, in the case of CML, highly effective treatment options. Progression to the more aggressive stages of the diseases limits effective treatment and significantly exacerbates patient prognosis. The mechanisms causing delay and disease progression are largely unknown. The later stages of these diseases are characterized by excessive build up of primitive cell types, indicating a disruption in the normal cell differentiation process that is commonly regulated through feedback from differentiated types. In this study, we propose a mechanism where mutated primitive cells produce a feedback interference signal that desensitizes them to a normal homeostatic feedback. Using a mathematical model, we show that this mechanism can account for the long delay period between occurrence of genetic changes and symptomatic onset characterized by fast growth of cancerous cell population. Finally, we explore novel concepts for potential treatment of chronic cancers.


Management of pulmonary hypertension in Down syndrome.
Children with Down syndrome (DS) are at greater risk of pulmonary arterial hypertension (PAH) than the general population, partly due to upper airway obstruction and congenital heart disease. We wished to review our management of PAH and suggest a protocol for the systematic management of these children. Children with DS and PAH were included as referred for assessment from March 2005 to May 2010. Twenty-five patients (13 boys) met inclusion criteria. The median age was 385 days (range, 106 to 5,734); mean tricuspid regurgitation jet was 3.5 (range, 2.7-4.8) m/s. At cardiac catheterisation, mean pulmonary artery mean pressure was 26 mmHg (range, 12 to 46), and mean pulmonary vascular resistance (PVR) was 4.14 U.m² (range, 1.20 to 12.43) at baseline. PVR fell to a mean of 2.68 U.m² (range, 0.38 to 10.69) with 20 ppm inhaled nitric oxide and 100% oxygen. Respiratory assessment included polysomnography (18), bronchoscopy (16), showing malacia (eight), adenotonsillar hypertrophy (eight) and floppy aryepiglottic folds (four). One lung biopsy showed plexogenic arteriopathy, and one was diagnosed with tracheo-oesophageal fistula. In order to manage this complex group of patients, a combined cardiological, respiratory and surgical approach was required. A protocol with cardiac catheterisation, blood tests and respiratory assessment is suggested for the management of pulmonary hypertension in these children.


Silicon surface passivation by PEDOT: PSS functionalized by SnO2 and TiO2 nanoparticles.
In this paper, we present a study of silicon surface passivation based on the use of spin-coated hybrid composite layers. We investigate both undoped poly(3,4-ethylenedioxythiophene)/poly-(styrenesulfonate) (PEDOT:PSS), as well as PEDOT:PSS functionalized with semiconducting oxide nanomaterials (TiO2 and SnO2). The hybrid compound was deposited at room temperature by spin coating-a potentially lower cost, lower processing time and higher throughput alternative compared with the commonly used vacuum-based techniques. Photoluminescence imaging was used to characterize the electronic properties of the Si/PEDOT:PSS interface. Good surface passivation was achieved by PEDOT:PSS functionalized by semiconducting oxides. We show that control of the concentration of semiconducting oxide nanoparticles in the polymer is crucial in determining the passivation performance. A charge carrier lifetime of about 275 μs has been achieved when using SnO2 nanoparticles at a concentration of 0.5 wt.% as a filler in the composite film. X-ray diffraction (XRD), scanning electron microscopy, high resolution transmission electron microscopy (HRTEM), energy dispersive x-ray in an SEM, and μ-Raman spectroscopy have been used for the morphological, chemical and structural characterization. Finally, a simple model of a photovoltaic device based on PEDOT:PSS functionalized with semiconducting oxide nanoparticles has been fabricated and electrically characterized.


Gastroesophageal reflux disease versus Helicobacter pylori infection as the cause of gastric carditis.
To explore the potential contributions of gastroesophageal reflux disease, as opposed to Helicobacter pylori infection, to the development of gastric carditis, we evaluated gastric carditis (using the criteria of the updated Sydney system for the classification of gastritis), clinical and morphologic features of esophagitis, and H. pylori infection (evaluation of Steiner stains) in biopsy specimens from the gastroesophageal squamocolumnar junction. We correlated clinical, endoscopic, and histologic features in an unselected group of 116 patients. Some degree of carditis was found in 107 (92%) of the patients. The mean age of the patients increased with increasing severity of carditis (P < .05). The various groups of patients with different degrees of carditis did not differ significantly in sex ratio, ethnic background, presence of obesity, percentage having symptoms of gastroesophageal reflux disease (such as heartburn, regurgitation, dysphagia, or odynophagia), endoscopic evidence of esophagitis and columnar epithelium in the distal esophagus, or histologic evidence of active esophagitis. The presence, however, of active gastritis and H. pylori infection in the distal stomach and/or in the cardia was significantly associated with carditis. In patients without carditis, H. pylori was not detected in any cardiac or distal gastric biopsy specimen. In contrast, H. pylori was demonstrated in gastric tissue samples (either from the cardia or distally) of patients with carditis, with the prevalence rate increasing with greater degrees of cardiac inflammation. The H. pylori prevalence rate was 12% in the group with mild carditis, 40% in those with moderate carditis, and 57% in patients with marked carditis (P = .0001). In summary, carditis is commonly found in patients with symptoms related to upper gastrointestinal diseases. From analysis of our study cohort, we concluded that carditis was significantly associated with H. pylori infection and active gastritis but not with symptoms or signs of gastroesophageal reflux disease. These findings suggest that carditis with histologic features similar to those of gastritis in the distal stomach was a sequel of H. pylori infection and represented a part of an H. pylori--associated gastric inflammation.


A hantavirus pulmonary syndrome (HPS) DNA vaccine delivered using a spring-powered jet injector elicits a potent neutralizing antibody response in rabbits and nonhuman primates.
Sin Nombre virus (SNV) and Andes virus (ANDV) cause most of the hantavirus pulmonary syndrome (HPS) cases in North and South America, respectively. The chances of a patient surviving HPS are only two in three. Previously, we demonstrated that SNV and ANDV DNA vaccines encoding the virus envelope glycoproteins elicit high-titer neutralizing antibodies in laboratory animals, and (for ANDV) in nonhuman primates (NHPs). In those studies, the vaccines were delivered by gene gun or muscle electroporation. Here, we tested whether a combined SNV/ANDV DNA vaccine (HPS DNA vaccine) could be delivered effectively using a disposable syringe jet injection (DSJI) system (PharmaJet, Inc). PharmaJet intramuscular (IM) and intradermal (ID) needle-free devices are FDA 510(k)-cleared, simple to use, and do not require electricity or pressurized gas. First, we tested the SNV DNA vaccine delivered by PharmaJet IM or ID devices in rabbits and NHPs. Both IM and ID devices produced high-titer anti-SNV neutralizing antibody responses in rabbits and NHPs. However, the ID device required at least two vaccinations in NHP to detect neutralizing antibodies in most animals, whereas all animals vaccinated once with the IM device seroconverted. Because the IM device was more effective in NHP, the Stratis(®) (PharmaJet IM device) was selected for follow-up studies. We evaluated the HPS DNA vaccine delivered using Stratis(®) and found that it produced high-titer anti-SNV and anti-ANDV neutralizing antibodies in rabbits (n=8/group) as measured by a classic plaque reduction neutralization test and a new pseudovirion neutralization assay. We were interested in determining if the differences between DSJI delivery (e.g., high-velocity liquid penetration through tissue) and other methods of vaccine injection, such as needle/syringe, might result in a more immunogenic DNA vaccine. To accomplish this, we compared the HPS DNA vaccine delivered by DSJI versus needle/syringe in NHPs (n=8/group). We found that both the anti-SNV and anti-ANDV neutralizing antibody titers were significantly higher (p-value 0.0115) in the DSJI-vaccinated groups than the needle/syringe group. For example, the anti-SNV and anti-ANDV PRNT50 geometric mean titers (GMTs) were 1,974 and 349 in the DSJI-vaccinated group versus 87 and 42 in the needle/syringe group. These data demonstrate, for the first time, that a spring-powered DSJI device is capable of effectively delivering a DNA vaccine to NHPs. Whether this HPS DNA vaccine, or any DNA vaccine, delivered by spring-powered DSJI will elicit a strong immune response in humans, requires clinical trials.


High rate of disease-related copy number variations in childhood onset schizophrenia.
Copy number variants (CNVs) are risk factors in neurodevelopmental disorders, including autism, epilepsy, intellectual disability (ID) and schizophrenia. Childhood onset schizophrenia (COS), defined as onset before the age of 13 years, is a rare and severe form of the disorder, with more striking array of prepsychotic developmental disorders and abnormalities in brain development. Because of the well-known phenotypic variability associated with pathogenic CNVs, we conducted whole genome genotyping to detect CNVs and then focused on a group of 46 rare CNVs that had well-documented risk for adult onset schizophrenia (AOS), autism, epilepsy and/or ID. We evaluated 126 COS probands, 69 of which also had a healthy full sibling. When COS probands were compared with their matched related controls, significantly more affected individuals carried disease-related CNVs (P=0.017). Moreover, COS probands showed a higher rate than that found in AOS probands (P<0.0001). A total of 15 (11.9%) subjects exhibited at least one such CNV and four of these subjects (26.7%) had two. Five of 15 (4.0% of the sample) had a 2.5-3 Mb deletion mapping to 22q11.2, a rate higher than that reported for adult onset (0.3-1%) (P<0.001) or autism spectrum disorder and, indeed, the highest rate reported for any clinical population to date. For one COS subject, a duplication found at 22q13.3 had previously only been associated with autism, and for four patients CNVs at 8q11.2, 10q22.3, 16p11.2 and 17q21.3 had only previously been associated with ID. Taken together, these findings support the well-known pleiotropic effects of these CNVs suggesting shared abnormalities early in brain development. Clinically, broad CNV-based population screening is needed to assess their overall clinical burden.


A second form of gonadotropin-releasing hormone (GnRH) with characteristics of chicken GnRH-II is present in the primate brain.
The primate brain was thought to contain only the GnRH known as mammalian GnRH (mGnRH). This study investigates whether a second form of GnRH exists within the primate brain. We found that brain extracts from adult stumptail and rhesus monkeys contained two forms of GnRH that were similar to mGnRH and chicken GnRH-II (cGnRH-II) based on the elution position of the peptides from HPLC and on cross-reactivity with antisera that are specific to mammalian or chicken GnRH-II in RIAs. The fetal brain of rhesus monkeys also contained mGnRH and a cGnRH-II-like peptide by the same criteria. Immunocytochemistry with a cGnRH-II-specific antiserum in adult and fetal rhesus monkeys showed immunopositive neurons generally scattered in the periaqueductal region of the midbrain, with a few positive cells in the posterior basal hypothalamus. Neurons immunopositive for cGnRH-II were fewer in number and smaller in size, with less defined nuclei and thinner neurites compared with those for mGnRH. Administration of synthetic cGnRH-II to adult rhesus monkeys resulted in a significant increase in the plasma LH concentration during the luteal phase of the menstrual cycle, but not during the midfollicular phase. We conclude that the primate brain contains mGnRH and a cGnRH-II-like molecule, although the function of the latter is unknown.


Structure of Sesquisabinene Synthase 1, a Terpenoid Cyclase That Generates a Strained [3.1.0] Bridged-Bicyclic Product.
The natural product sesquisabinene is a key component of the fragrant essential oil of the sandalwood tree, currently valued at $5,000/L. Sesquisabinene contains a highly strained [3.1.0] bicyclic ring system and is generated from farnesyl diphosphate in a reaction catalyzed by a class I terpenoid cyclase. To understand how the enzyme directs the formation of a strained hydrocarbon ring system, we now report the X-ray crystal structure of sesquisabinene synthase 1 (SQS1) from the Indian sandalwood tree ( Santalum album). Specifically, we report the structure of unliganded SQS1 at 1.90 Å resolution and the structure of its complex with three Mg2+ ions and the inhibitor ibandronate at 2.10 Å resolution. The bisphosphonate group of ibandronate coordinates to all three metal ions and makes hydrogen bond interactions with basic residues at the mouth of the active site. These interactions are similarly required for activation of the substrate diphosphate group to initiate catalysis, although partial occupancy binding of the Mg2+B ion suggests that this structure represents the penultimate metal coordination complex just prior to substrate activation. The structure of the liganded enzyme enables a precise definition of the enclosed active site contour that serves as a template for the cyclization reaction. This contour is very product-like in shape and readily fits an extended conformation of sesquisabinene and its precursor, the homobisabolyl cation. Structural comparisons of SQS1 with epi-isozizaene synthase mutants that also generate sesquisabinene suggest that [3.1.0] ring formation is not dependent on the isoprenoid tail conformation of the homobisabolyl cation.


[Repetitive strain injury (RSI); a report from the Health Council of the Netherlands].
The Health Council of the Netherlands has published its advisory report on Repetitive Strain Injury (RSI). The report provides clear information on the state of this syndrome, including the definition of the problem, the epidemiology, various hypothetical pathophysiological mechanisms, occupational and personal risk factors, and possible methods of treatment. The council states that with regard to the last aspect, too few data are available to draw any conclusion as to the most promising therapy. Nevertheless, patients should get consistent advice from their GP or company doctor. The council emphasises that encouraging physical exercise and eliminating any possible causative strain should be part of an integrated approach, embracing work-related psychosocial and personal issues. Of particular interest is the council's advice to prevent RSI by improving the physical condition and by selective training of muscle function. The report recommends that more research be carried out in order to provide insight into the effectiveness of the treatment of RSI.


The Relationship between Perceived Stress and Marital Satisfaction in Couples with Infertility: Actor-Partner Interdependence Model.
Infertility, one of life's great stressors, may adversely affect marital satisfaction. No studies have investigated the relationship between perceived stress and marital satisfaction at the dyadic level. The current study assessed the actor and partner effects of perceived stress on marital satisfaction in husband-wife dyads using an innovative dyadic analysis approach, the Actor-Partner Interdependence Model (APIM). In this cross-sectional study, we recruited a total of 141 infertile couples. Marital satisfaction and stress were assessed using the ENRICH Marital Satisfaction Scale (EMS Scale) and Perceived Stress Scale-4 Item (PSS-4), respectively. Dyadic data have been analysed by the APIM approach, with distinguishable dyads. In this approach, actor effect is the impact of a person's perceived stress on his/her own marital satisfaction. Partner effect is the impact of a person's perceived stress on the partner's marital satisfaction. Both men and women's perceived stress exhibited an actor effect on their marital satisfaction (β=-0.312, P<0.001, β=-0.405, P<0.001, respectively). Women's perceived stress had a negative relationship to the marital satisfaction of their partner (β=-0.174, P=0.040). Although the partner effect of men's perceived stress on woman's marital satisfaction was not significant (β=-0.138, P=0.096), women whose husbands had higher levels of stress were more likely to have poorer marital satisfaction. Both actor and partner effects of perceived stress on marital satisfaction were similar among men and their wives. The findings of this study have highlighted that marital satisfaction in patients with infertility was influenced by not only their own perceived stress, but also their spouses' perceived stresses. Therefore, psychological interventions that target a reduction in perceived stress and enhancement of marital satisfaction in the context of infertility should treat the couple as a unit.


Human T cell leukemia virus-I (HTLV-I) Tax-mediated apoptosis in activated T cells requires an enhanced intracellular prooxidant state.
We have shown that an estradiol-dependent activation of human T cell leukemia virus-I Tax leads to the inhibition of cell proliferation and to the induction of apoptosis. The present study demonstrates that a hormone-dependent activation of Tax promotes an enhanced prooxidant state in stably transfected Jurkat cells as measured by changes in the intracellular levels of glutathione and H2O2; these changes are followed by apoptotic cell death. Additional stimulation of the CD3/TCR pathway enhances the oxidative and apoptotic effects. Both Tax-mediated apoptosis and oxidative stress can be potently suppressed by antioxidants, as is seen with the administration of recombinant thioredoxin (adult T cell leukemia-derived factor) or pyrrolidine dithiocarbamate. Hormone-induced Tax activation induces a long-lasting activation of NF-kappaB, which is a major target of reactive oxygen intermediates. The long-term exposure of Jurkat cells to hormone eventually results in a selection of cell clones that have lost Tax activity. A subsequent transfection of these apparently "nonresponsive" clones allows the recovery of Tax responses in these cells. Our observations indicate that changes in the intracellular redox status may be a determining factor in Tax-mediated DNA damage, apoptosis, and selection against the long-term expression of Tax function.


H2AFY promoter deletion causes PITX1 endoactivation and Liebenberg syndrome.
Structural variants (SVs) affecting non-coding cis-regulatory elements are a common cause of congenital limb malformation. Yet, the functional interpretation of these non-coding variants remains challenging. The human Liebenberg syndrome is characterised by a partial transformation of the arms into legs and has been shown to be caused by SVs at the PITX1 locus leading to its misregulation in the forelimb by its native enhancer element Pen. This study aims to elucidate the genetic cause of an unsolved family with a mild form of Liebenberg syndrome and investigate the role of promoters in long-range gene regulation. Here, we identify SVs by whole genome sequencing (WGS) and use CRISPR-Cas9 genome editing in transgenic mice to assign pathogenicity to the SVs. In this study, we used WGS in a family with three mildly affected individuals with Liebenberg syndrome and identified the smallest deletion described so far including the first non-coding exon of H2AFY. To functionally characterise the variant, we re-engineered the 8.5 kb deletion using CRISPR-Cas9 technology in the mouse and showed that the promoter of the housekeeping gene H2afy insulates the Pen enhancer from Pitx1 in forelimbs; its loss leads to misexpression of Pitx1 by the pan-limb activity of the Pen enhancer causing Liebenberg syndrome. Our data indicate that housekeeping promoters may titrate promiscuous enhancer activity to ensure normal morphogenesis. The deletion of the H2AFY promoter as a cause of Liebenberg syndrome highlights this new mutational mechanism and its role in congenital disease.


Subdivision design and stewardship affect bird and mammal use of conservation developments.
Developing effective tools for conservation on private lands is increasingly important for global biodiversity conservation; private lands are located in more productive and biologically diverse areas, and they face accelerated rates of land conversion. One strategy is conservation development (CD) subdivisions, which cluster houses in a small portion of a property and preserve the remaining land as protected open space. Despite widespread use, the characteristics that make CD more or less effective at achieving biodiversity conservation are not well understood. We investigated CD's ability to successfully protect animal populations by examining bird and mammal occurrences in 14 CD subdivisions and four undeveloped areas (range: 14-432 ha) in northern Colorado, USA. Using point count and camera trap data in an occupancy modeling framework, we evaluated the relative importance of nine subdivision design factors (e.g., housing density, proportion of CD protected) and 14 stewardship factors (e.g., presence of livestock, percent native vegetation cover) in influencing the overall community composition and the probability of use by 16 birds and six mammals. We found that habitat use by 75% of birds and 83% of mammals was associated with design characteristics that maximized the natural or undisturbed land area both within and near the development (e.g., proportion of CD protected, total area of protected open space, proportion of natural land cover in the surrounding landscape). These factors were also associated with an increasing dominance of human-sensitive bird species, larger-bodied mammals, and mammals with larger home ranges. Habitat use by birds was also influenced by local land use composition and quality, and use by several bird and mammal species decreased with increased localized disturbances. We found few differences in habitat use between sampling sites in undeveloped areas and in CD subdivisions. These similarities indicate that, if CDs are large enough or located within a matrix of undeveloped land, they can provide habitat that supports similar use patterns as protected areas without housing development. By incorporating characteristics that promote the persistence of sensitive birds and mammals on private lands, CDs have potential to preserve native biodiversity in areas threatened by expanding residential development.


Effects of transmission of Y chromosome AZFc deletions.
Deletions of specific regions on the Y chromosome cause male infertility. Recent advances in infertility treatment allow Y chromosome deletions to be transmitted to male offspring with the assumption that there will be no clinical consequences other than infertility in adult life. We screened 12 patients, who had a 45X/46XY karyotype and presented with Turner stigmata or sexual ambiguities, or both, for Y chromosome microdeletions with PCR. A third of these patients had Y chromosome microdeletions of distal Yq, the most common microdeletion seen in infertile men with azoospermia or severe oligozoospermia. Transmission of Y chromosome microdeletions could potentially have severe clinical consequences other than male infertility, such as the development of sexual ambiguities and Turner stigmata.


Infections in travellers returning to Turkey from the Arabian peninsula: a retrospective cross-sectional multicenter study.
Mass gatherings pooling people from different parts of the world-the largest of which is to Mecca, Saudi Arabia, for Hajj-may impose risks for acquisition and dissemination of infectious diseases. A substantial number of pilgrims to Hajj and Umrah are Turkish citizens (456,000 in 2014) but data are lacking on scale of the problem. We did a retrospective cross-sectional multicenter study in Turkey to explore the range of infections among inpatients who had recently returned from the Arabian Peninsula. Our inclusion criteria were patients who had acquired an infection during their trip to an Arabian Peninsula country, or who became symptomatic within 1 week of their return. The data were collected retrospectively for January 1, 2013 and March 1, 2015. 185 Turkish patients were recruited to the study across 15 referral centers with travel associated infectious diseases after returning from Arabian Peninsula countries (predominantly Saudi Arabia 163 [88.1 %] for religious purposes 162 [87.5 %]). Seventy four (40.0 %) of them were ≥ 65 years old with numerous comorbidities including diabetes (24.3 %) and COPD (14.1 %). The most common clinical diagnosis was respiratory tract infections (169 [91.5 %]), followed by diarrheal diseases (13 [7 %]), and there was one case of MERS-CoV. Patients spent a median of 5 (3-7) days as hospital inpatients and overall mortality was 1.1 %. Returning travellers from the Arabian Peninsula present as inpatients with a broad range of infectious diseases similar to common community acquired infections frequently seen in daily medical practices in Turkey.


AUDIT-C alcohol screening results and postoperative inpatient health care use.
Alcohol screening scores ≥5 on the Alcohol Use Disorders Identification Test-Consumption (AUDIT-C) up to a year before surgery have been associated with postoperative complications, but the association with postoperative health care use is unknown. This study evaluated whether AUDIT-C scores in the year before surgery were associated with postoperative hospital length of stay, total ICU days, return to the operating room, and hospital readmission. This cohort study included male Veterans Affairs patients who completed the AUDIT-C on mailed surveys (October 2003 through September 2006) and were hospitalized for nonemergent noncardiac major operations in the following year. Postoperative health care use was evaluated across 4 AUDIT-C risk groups (scores 0, 1 to 4, 5 to 8, and 9 to 12) using linear or logistic regression models adjusted for sociodemographics, smoking status, surgical category, relative value unit, and time from AUDIT-C to surgery. Patients with AUDIT-C scores indicating low-risk drinking (scores 1 to 4) were the referent group. Adjusted analyses revealed that among eligible surgical patients (n = 5,171), those with the highest AUDIT-C scores (ie, 9 to 12) had longer postoperative hospital length of stay (5.8 [95% CI, 5.0-6.7] vs 5.0 [95% CI, 4.7-5.3] days), more ICU days (4.5 [95% CI, 3.2-5.8] vs 2.8 [95% CI, 2.6-3.1] days), and increased probability of return to the operating room (10% [95% CI, 6-13%] vs 5% [95% CI, 4-6%]) in the 30 days after surgery, but not increased hospital readmission within 30 days postdischarge, relative to the low-risk group. AUDIT-C screening results could be used to identify patients at risk for increased postoperative health care use who might benefit from preoperative alcohol interventions.


A meta-analysis of the effect of estrogen replacement therapy on the risk of breast cancer.
To quantify the effect of estrogen replacement therapy on breast cancer risk, we combined dose-response slopes of the relative risk of breast cancer against the duration of estrogen use across 16 studies. Using this summary dose-response slope, we calculated the proportional increase in risk of breast cancer for each year of estrogen use. For women who experienced any type of menopause, risk did not appear to increase until after at least 5 years of estrogen use. After 15 years of estrogen use, we found a 30% increase in the risk of breast cancer (relative risk, 1.3; 95% confidence interval [CI], 1.2 to 1.6). The increase in risk was largely due to results of studies that included premenopausal women or women using estradiol (with or without progestin), studies for which the estimated relative risk was 2.2 (CI, 1.4 to 3.4) after 15 years. Among women with a family history of breast cancer, those who had ever used estrogen replacement had a significantly higher risk (3.4; CI, 2.0 to 6.0) than those who had not (1.5; CI, 1.2 to 1.7).


The crystal structure of human pyrin b30.2 domain: implications for mutations associated with familial Mediterranean fever.
The inherited autoinflammatory syndrome familial Mediterranean fever (FMF) is characterized by recurrent episodes of fever, which are independent of any bacterial or viral infections. This disease is associated with point mutations in the mefv gene product pyrin. Although the precise molecular functions of pyrin are unknown, it seems to be involved in the maturation and secretion of interleukin-1beta. Approximately two thirds of all FMF-associated mutations cluster in the C-terminal B30.2 domain of pyrin. To investigate the molecular consequences of FMF-associated mutations, we determined the crystal structure of the pyrin B30.2 domain at 1.35-A resolution. The comparison with other B30.2/ligand complex structures revealed a shallow cavity, which seems to be involved in binding the pyrin ligand. The bottom of this cavity is covered mainly with hydrophobic amino acids, suggesting that pyrin recognizes its ligand by hydrophobic contacts and surface complementarities. FMF-associated mutations cluster around two sites on the B30.2 surface. Approximately two thirds, including those mutations with the most severe disease outcomes, are observed in the vicinity of the predicted peptide binding site, suggesting that they will have a direct impact on ligand binding. A second mutational hot spot was observed on the opposite side of the B30.2 domain in the neighbourhood of its artificial N-terminus. Although most FMF-associated mutations are solvent exposed, several will modify the main-chain conformation of loops. The experimental crystal structure of the pyrin B30.2 domain serves as a basis for an accurate modelling of these mutations.


Measuring the mechanical properties of individual human blood cells.
The largest human blood cells--the red cells (erythrocytes) and white cells (leukocytes)--must undergo a significant amount of deformation as they squeeze through the smallest vessels of the circulation and the small openings between bone, vessel and tissue. This ability to deform in response to external forces shows that cells exhibit material behavior and behave as either elastic solids or viscous liquids. The question then is "how can we measure the deformation and flow of something as small as a blood cell and what kinds of constitutive equations describe cellular deformation"? In this paper the use of the micropipet to measure red cell and white cell, especially neutrophil, deformation will be described and the viscoelastic models used to describe the deformation behavior of red cell membrane and neutrophil cytoplasm will be discussed. Values for the elasticities of a red cell membrane subjected to shear, area expansion and bending will be given. The viscosity of red cell membrane in shear will also be discussed. Finally, the cortical tension of the neutrophil and the Newtonian and Maxwell models used to characterize its apparent viscosity will be discussed even though neither is wholly successful in describing the viscous behavior of the neutrophil. Thus, alternate models will be suggested.


Loss-of-function mutations in the histone methyltransferase EZH2 promote chemotherapy resistance in AML.
Chemotherapy resistance is the main impediment in the treatment of acute myeloid leukaemia (AML). Despite rapid advances, the various mechanisms inducing resistance development remain to be defined in detail. Here we report that loss-of-function mutations (LOF) in the histone methyltransferase EZH2 have the potential to confer resistance against the chemotherapeutic agent cytarabine. We identify seven distinct EZH2 mutations leading to loss of H3K27 trimethylation via multiple mechanisms. Analysis of matched diagnosis and relapse samples reveal a heterogenous regulation of EZH2 and a loss of EZH2 in 50% of patients. We confirm that loss of EZH2 induces resistance against cytarabine in the cell lines HEK293T and K562 as well as in a patient-derived xenograft model. Proteomics and transcriptomics analysis reveal that resistance is conferred by upregulation of multiple direct and indirect EZH2 target genes that are involved in apoptosis evasion, augmentation of proliferation and alteration of transmembrane transporter function. Our data indicate that loss of EZH2 results in upregulation of its target genes, providing the cell with a selective growth advantage, which mediates chemotherapy resistance.


Prevalence and Impact on Weaning of Pleural Effusion at the Time of Liberation from Mechanical Ventilation: A Multicenter Prospective Observational Study.
Pleural effusion is frequent in intensive care unit patients, but its impact on the outcome of weaning remains unknown. In a prospective study performed in three intensive care units, pleural ultrasound was performed at the first spontaneous breathing trial to detect and quantify pleural effusion (small, moderate, and large). Weaning failure was defined by a failed spontaneous breathing trial and/or extubation requiring any form of ventilatory support within 48 h. The primary endpoint was the prevalence of pleural effusion according to weaning outcome. Pleural effusion was detected in 51 of 136 (37%) patients and was quantified as moderate to large in 18 (13%) patients. As compared to patients with no or small pleural effusion, their counterparts were more likely to have chronic renal failure (39 vs. 7%; P = 0.01), shock as the primary reason for admission (44 vs. 19%; P = 0.02), and a greater weight gain (+4 [0 to 7] kg vs. 0 [-1 to 5] kg; P = 0.02). The prevalence of pleural effusion was similar in weaning success and weaning failure patients (odds ratio, 1.23; 95% CI, 0.61 to 2.49; P = 0.56), as was the prevalence of moderate to large pleural effusion (odds ratio, 0.89; 95% CI, 0.33 to 2.41; P = 1.00). Duration of mechanical ventilation and intensive care unit length of stay were similar between patients with no or small pleural effusion and those with moderate to large pleural effusion. Significant pleural effusion was observed in 13% of patients at the time of liberation from mechanical ventilation and was not associated with an alteration of weaning outcome. (ANESTHESIOLOGY 2017; 126:1107-15).


Impact of congestive heart failure in patients with non-ST-segment elevation acute coronary syndromes.
The presence of congestive heart failure (CHF) has been associated with treatment disparities and worse outcomes in patients with ST-segment elevation myocardial infarction, but the incidence and effect of CHF in patients with non-ST-segment elevation acute coronary syndromes (NSTE ACSs) has not been well characterized. We evaluated 45,744 patients with NSTE ACS (positive cardiac markers and/or ischemic ST-segment changes) who were treated at 424 hospitals in the CRUSADE Quality Improvement Initiative between March 2000 and March 2003. Treatment patterns and in-hospital outcomes in patients with signs of CHF on presentation and those who developed in-hospital CHF were compared with those in patients without CHF. In total, 10,398 patients (22.7%) had signs of CHF on presentation, and 1,664 patients (3.6%) later developed in-hospital CHF. Compared with patients without CHF, early (<24 hours from presentation) medications and invasive cardiac procedures were used less often in patients with signs of CHF on presentation. Likewise, patients with in-hospital CHF were less likely than those without CHF to receive acute antiplatelet agents and undergo cardiac catheterization but more likely to receive acute beta blockers, angiotensin-converting enzyme inhibitors, and heparin and to undergo coronary artery bypass grafting. Adjusted mortality was higher in patients with signs of CHF on presentation (odds ratio 2.64, 95% confidence interval 2.31 to 3.01) and those with in-hospital CHF (odds ratio 4.93, 95% confidence interval 4.05 to 5.99) than in patients without CHF. In conclusion, CHF occurs frequently in patients with NSTE ACS but is associated with less aggressive treatment and a higher risk of mortality. Further study is needed to determine the causes of these treatment differences and the optimal therapeutic approach for patients with NSTE ACS and concomitant CHF.


ST2 as checkpoint target for colorectal cancer immunotherapy.
Immune checkpoint blockade immunotherapy delivers promising clinical results in colorectal cancer (CRC). However, only a fraction of cancer patients develop durable responses. The tumor microenvironment (TME) negatively impacts tumor immunity and subsequently clinical outcomes. Therefore, there is a need to identify other checkpoint targets associated with the TME. Early-onset factors secreted by stromal cells as well as tumor cells often help recruit immune cells to the TME, among which are alarmins such as IL-33. The only known receptor for IL-33 is stimulation 2 (ST2). Here we demonstrated that high ST2 expression is associated with poor survival and is correlated with low CD8+ T cell cytotoxicity in CRC patients. ST2 is particularly expressed in tumor-associated macrophages (TAMs). In preclinical models of CRC, we demonstrated that ST2-expressing TAMs (ST2+ TAMs) were recruited into the tumor via CXCR3 expression and exacerbated the immunosuppressive TME; and that combination of ST2 depletion using ST2-KO mice with anti-programmed death 1 treatment resulted in profound growth inhibition of CRC. Finally, using the IL-33trap fusion protein, we suppressed CRC tumor growth and decreased tumor-infiltrating ST2+ TAMs. Together, our findings suggest that ST2 could serve as a potential checkpoint target for CRC immunotherapy.


Fatty acid regulation of gene expression: a genomic explanation for the benefits of the mediterranean diet.
The development of obesity and associated insulin resistance involves a multitude of gene products, including proteins involved in lipid synthesis and oxidation, thermogenesis, and cell differentiation. The genes encoding these proteins are in essence the blueprints that we have inherited from our parents. However, what determines the way in which blueprints are interpreted is largely dictated by a collection of environmental factors. The nutrients we consume are among the most influential of these environmental factors. During the early stages of evolutionary development, nutrients functioned as primitive hormonal signals that allowed the early organisms to turn on pathways of synthesis or storage during periods of nutrient deprivation or excess. As single-cell organisms evolved into complex life forms, nutrients continued to be environmental factors that interacted with hormonal signals to govern the expression of genes encoding proteins involved in energy metabolism, cell differentiation, and cell growth. Nutrients govern the tissue content and activity of different proteins by functioning as regulators of gene transcription, nuclear RNA processing, mRNA degradation, and mRNA translation, as well as functioning as posttranslational modifiers of proteins. One dietary constituent that has a strong influence on cell differentiation, growth, and metabolism is fat. The fatty acid component of dietary lipid not only influences hormonal signaling events by modifying membrane lipid composition, but fatty acids have a very strong direct influence on the molecular events that govern gene expression. In this review, we discuss the influence that (n-9), (n-6), and (n-3) fatty acids exert on gene expression in the liver and skeletal muscle and the impact this has on intra- and interorgan partitioning of metabolic fuels.


Structural and energetic basis for hybridization limits in high-density DNA monolayers.
High-density monolayers (HDMs) of single-strand (ss) DNA are important nanoscale platforms for the fabrication of sensors and for mechanistic studies of enzymes on surfaces. Such systems can be used, for example, to monitor gene expression, and for the construction of more complex nanodevices via selective hybridization with the complementary oligos dissolved in solution. In this framework, controlling HDM hybridization is essential to control the final properties. Different studies demonstrate that at the typical density of ≈10(13) molecules per cm(2) no more than ≈30-40% of the HDM ssDNA is successfully hybridized. Until now, however, the origin of the HDM hybridization limit has remained unclear. In this work, molecular dynamics (MD) simulations of HDM systems with variable hybridization reveal that, independently of other experimental parameters, the effective hybridization for a HDM of this density is intrinsically limited by molecular and electrostatic crowding. A detailed structural analysis of the HDM model shows good agreement with our atomic force microscopy (AFM) experiments, and provides further insight into the steric hindrance behaviour and time-resolved surface topography of these nanostructured systems. The explicit relationship proposed between structural crowding and limited HDM hybridization offers a rationale to control the final properties of HDM-based nanodevices.


The relationship of three cortical regions to an information-processing model.
This research tests a model of the computational role of three cortical regions in tasks like algebra equation solving. The model assumes that there is a left parietal region-of-interest (ROI) where the problem expression is represented and transformed, a left prefrontal ROI where information for solving the task is retrieved, and a motor ROI where hand movements to produce the answer are programmed. A functional magnetic resonance imaging (fMRI) study of an abstract symbol-manipulation task was performed to articulate the roles of these three regions. Participants learned to associate words with instructions for transforming strings of letters. The study manipulated the need to retrieve these instructions, the need to transform the strings, and whether there was a delay between calculation of the answer and the output of the answer. As predicted, the left parietal ROI mainly reflected the need for a transformation and the left prefrontal ROI the need for retrieval. Homologous right ROIs showed similar but weaker responses. Neither the prefrontal nor the parietal ROIs responded to delay, but the motor ROI did respond to delay, implying motor rehearsal over the delay. Except for the motor ROI, these patterns of activity did not vary with response hand. In an ACT-R model, it was shown that the activity of an imaginal buffer predicted the blood oxygen level-dependent (BOLD) response of the parietal ROI, the activity of a retrieval buffer predicted the response of the prefrontal ROI, and the activity of a manual buffer predicted the response of the motor ROI.


Prescription Stimulants' Effects on Healthy Inhibitory Control, Working Memory, and Episodic Memory: A Meta-analysis.
The use of prescription stimulants to enhance healthy cognition has significant social, ethical, and public health implications. The large number of enhancement users across various ages and occupations emphasizes the importance of examining these drugs' efficacy in a nonclinical sample. The present meta-analysis was conducted to estimate the magnitude of the effects of methylphenidate and amphetamine on cognitive functions central to academic and occupational functioning, including inhibitory control, working memory, short-term episodic memory, and delayed episodic memory. In addition, we examined the evidence for publication bias. Forty-eight studies (total of 1,409 participants) were included in the analyses. We found evidence for small but significant stimulant enhancement effects on inhibitory control and short-term episodic memory. Small effects on working memory reached significance, based on one of our two analytical approaches. Effects on delayed episodic memory were medium in size. However, because the effects on long-term and working memory were qualified by evidence for publication bias, we conclude that the effect of amphetamine and methylphenidate on the examined facets of healthy cognition is probably modest overall. In some situations, a small advantage may be valuable, although it is also possible that healthy users resort to stimulants to enhance their energy and motivation more than their cognition.


Inhibition of a class C beta-lactamase by a specific phosphonate monoester.
A phosphonate monoester, m-carboxyphenyl phenylacetamidomethylphosphonate, has been found to be a specific inhibitor of the class C beta-lactamase of Enterobacter cloacae P99. Inactivation is rapid (10(3) per second per molar concentration) and reactivation very slow (2.2 X 10(-6) per second). Apparently concerted with the inactivation, one equivalent (with respect to the enzyme) of m-hydroxybenzoate is released. Reactivation is accelerated by hydroxylamine and benzohydroxamate. This suggests that the loss of enzyme activity is due to phosphonylation of an active site functional group. This discovery holds the promise of a new general class of beta-lactamase inhibitors and, perhaps, antibiotics.


Modeling the interferometric radius measurement using Gaussian beam propagation.
We model the interferometric radius measurement using Gaussian beam propagation to identify biases in the measurement due to using a simple geometric ray-trace model instead of the more complex Gaussian model. The radius measurement is based on using an interferometer to identify the test part's position when it is at two null locations, and the distance between the positions is an estimate of the part's radius. The null condition is observed when there is no difference in curvature between the reflected reference and the test wavefronts, and a Gaussian model will provide a first-order estimate of curvature changes due to wave propagation and therefore changes to the radius measurement. We show that the geometric ray assumption leads to radius biases (errors) that are a strong function of the test part radius and increase as the radius of the part decreases. We tested for a bias for both microscaled (<1 mm) and macroscaled parts. The bias is of the order of parts in 10(5) for micro-optics with radii a small fraction of a millimeter and much smaller for macroscaled optics. The amount of bias depends on the interferometer configuration (numerical aperture, etc.), the nominal radius of the test part, and the distances in the interferometer.


Abdominal obesity, sarcopenia, and osteoporosis are associated with frailty in men living with and without HIV.
The relationships between frailty and body composition in older adults with HIV infection are poorly understood. We sought to describe associations between frailty and measures of body composition among adult men with HIV and without HIV. Men with and without HIV (age 50-69 years) in the Multicenter AIDS Cohort Study (MACS) Bone Strength Substudy were included if evaluated for frailty (by Fried phenotype) and body composition [BMI, waist circumference, abdominal visceral (VAT) and subcutaneous (SAT) adipose tissue, sarcopenia, and osteopenia/osteoporosis]. All participants with HIV infection were on antiretroviral therapy. Multivariate multinomial logistic regression models were used to determine associations of frailty with body composition. A total of 399 men, including 199 men with HIV and 200 men without HIV, both with median age 60 years, constituted our study population. Frailty prevalence was 16% (men with HIV) vs. 8% (men without HIV). HIV serostatus was associated with a 2.43 times higher odds of frailty (P = 0.01). Higher waist circumference, VAT, sarcopenia, and femoral neck osteoporosis were associated with increased odds of frailty (aOR 4.18, 4.45, 4.15, and 13.6, respectively, and all P < 0.05); BMI and SAT were not. None of these measures presented a differential association with frailty by HIV serostatus (all P > 0.20). Higher abdominal obesity and sarcopenia were associated with frailty among men with and without HIV. Assessment of these body composition parameters may help detect frailty in the clinical setting.


Scratching behavior and Fos expression in superficial dorsal horn elicited by protease-activated receptor agonists and other itch mediators in mice.
Protease-activated receptor (PAR)-2 and PAR-4 are implicated in nonhistaminergic itch. We investigated dose dependence, tachyphylaxis, and cross-tachyphylaxis of itch-associated scratching elicited by intradermal injections of PAR-2 and PAR-4 agonists, serotonin (5-hydroxytryptamine, 5-HT), and histamine in ICR mice, as well as mu-opioid modulation of PAR-2 agonist-evoked scratching. Each agent elicited dose-related increases in scratch bouts. Scratching elicited by the PAR-4 agonist and histamine both exhibited significant tachyphylaxis but no cross-tachyphylaxis with each other. Scratching evoked by 5-HT did not exhibit significant tachyphylaxis but did exhibit significant cross-tachyphylaxis to scratching evoked by the PAR-2 and PAR-4 agonists and histamine. Naltrexone and high-dose morphine (10 mg/kg) attenuated PAR-2 agonist-evoked scratching, whereas lower dose morphine (1 mg/kg) had no effect. High-dose morphine also significantly increased circling behavior, which may have interfered with scratching. The PAR-2 agonist and 5-HT produced overlapping distributions of Fos-like immunoreactivity in the superficial dorsal horn. These results indicate that PAR-2 and PAR-4 agonists, histamine, and 5-HT elicit itch-related scratching and activate superficial dorsal horn neurons that may participate in scratch reflex and ascending itch signaling pathways.


Health care input constraints and cost effectiveness analysis decision rules.
Results of cost effectiveness analyses (CEA) studies are most useful for decision makers if they face only one constraint: the health care budget. However, in practice, decision makers wishing to use the results of CEA studies may face multiple resource constraints relating to, for instance, constraints in health care inputs such as a shortage of skilled labour. The presence of multiple resource constraints influences the decision rules of CEA and limits the usefulness of traditional CEA studies for decision makers. The goal of this paper is to illustrate how results of CEA can be interpreted and used in case a decision maker faces a health care input constraint. We set up a theoretical model describing the optimal allocation of the health care budget in the presence of a health care input constraint. Insights derived from that model were used to analyse a stylized example based on a decision about a surgical robot as well as a published cost effectiveness study on eye care services in Zambia. Our theoretical model shows that applying default decision rules in the presence of a health care input constraint leads to suboptimal decisions but that there are ways of preserving the traditional decision rules of CEA by reweighing different cost categories. The examples illustrate how such adjustments can be made, and makes clear that optimal decisions depend crucially on such adjustments. We conclude that it is possible to use the results of cost effectiveness studies in the presence of health care input constraints if results are properly adjusted.


Intervertebral disc autografting in a bipedal animal model.
Fusion of the spine while restoring stability of the spinal segment, fails to preserve spinal mobility. Long term complications of accelerated degeneration in the neighboring segments have been reported. The present study explores the possibility of intervertebral disc autografting in a bipedal animal model by isolating a lumbar disc together with the adjacent end plates and repositioning it with minimal internal fixation. Fourteen Rhesus monkeys were sacrificed at 2, 4, 6, and 12 months after surgery and the grafted discs were examined radiologically, biochemically, pathologically, and biomechanically. Healing of the bony end plate was seen between 2 to 4 months postoperatively. There was early loss of disc height at 2 and 4 months but there was a suggestion of some reconstitution up to 12 months. There was minimal evidence of gross degeneration at all stages. Gradual loss of water content was found in the annulus and the nucleus. The nucleus pulposus seemed to be able to reaccumulate proteoglycan after an initial drop in the first 4 months. There was significant increase in hydroxyproline content in the annulus fibrosus and the nucleus pulposus. Biomechanically, the grafted disc showed hypermobility in the first 4 months but gradually became stabilized with time. Results from this study suggested that a fresh intervertebral disc autograft could survive a period of ischemia. Although the physiology of the disc was deranged, it was able to preserve a certain degree of segmental mobility without sacrificing stability. Further studies are required to validate these results and the field of disc allografting should be explored.


A Pilot Study to Assess Adenosine 5'-triphosphate Metabolism in Red Blood Cells as a Drug Target for Potential Cardiovascular Protection.
To study the effect of exercise preconditioning on adenosine 5'triphosphate (ATP) metabolism in red blood cells and cardiovascular protection against injury induced by isoproterenol in vivo. Male Sprague Dawley rats (SDR) were each exercised on a treadmill for 15 minutes at 10 m/min and 10% grade (n = 7) (LowEx), or 14 m/min and 22% grade (n = 8) (VigEx). Two hours after the exercise, each rat received a single dose of isoproterenol (30 mg/kg) by subcutaneous (sc) injection. Two separate groups of SDR were used as control: One received no exercise (n = 10) (NoEx) and the other received no exercise and no isoproterenol (n = 11) (NoIso). Serial blood samples were collected over 5 hours for measurement of ATP and its catabolites by a validated HPLC. Hemodynamic recording was collected continuously for the duration of the experiment. Data were analysed using ANOVA and t-tests and difference considered significant at p < 0.05. Exercise pre-conditioning (both LowEx and VigEx) reduced mortality after isoproterenol from 50% to < 30% (p > 0.05). It attenuated the rebound in blood pressure significantly (p < 0.05 between NoEx vs VigEx), attenuated the increase of RBC adenosine 5'-monophosphate (AMP) concentrations induced by isoproterenol, and also decreased the breakdown of ATP to AMP in the RBC (p < 0.05 vs NoEx). Exercise pre-conditioning decreased the blood pressure rebound and also breakdown of ATP in RBC after isoproterenol which may be exploited further as a drug target for cardiovascular protection and prevention.


Divided attention limits perception of 3-D object shapes.
Can one perceive multiple object shapes at once? We tested two benchmark models of object shape perception under divided attention: an unlimited-capacity and a fixed-capacity model. Under unlimited-capacity models, shapes are analyzed independently and in parallel. Under fixed-capacity models, shapes are processed at a fixed rate (as in a serial model). To distinguish these models, we compared conditions in which observers were presented with simultaneous or sequential presentations of a fixed number of objects (The extended simultaneous-sequential method: Scharff, Palmer, & Moore, 2011a, 2011b). We used novel physical objects as stimuli, minimizing the role of semantic categorization in the task. Observers searched for a specific object among similar objects. We ensured that non-shape stimulus properties such as color and texture could not be used to complete the task. Unpredictable viewing angles were used to preclude image-matching strategies. The results rejected unlimited-capacity models for object shape perception and were consistent with the predictions of a fixed-capacity model. In contrast, a task that required observers to recognize 2-D shapes with predictable viewing angles yielded an unlimited capacity result. Further experiments ruled out alternative explanations for the capacity limit, leading us to conclude that there is a fixed-capacity limit on the ability to perceive 3-D object shapes.


Exposure and reorganization: the what and how of effective psychotherapy.
Even though the effectiveness of psychotherapy is generally acknowledged, researchers are yet to agree on a plausible explanation for this effectiveness or on possible mechanisms of change that are activated by psychotherapy. To enhance developments in these areas some researchers have called for a focus on treatment principles rather than treatment techniques. In this respect, the technique of exposure is instructive. Despite its common use with anxiety disorders and the successful outcomes it produces, it has only recently been considered as a treatment for other disorders. By focussing on the underlying principles of exposure it is possible to consider exposure as a transdiagnostic component of successful psychotherapies. Understanding exposure from the perspective of Perceptual Control Theory (PCT) enables the identification of a functional, rather than a conceptual or statistical, mechanism of change. Functionally, exposure can be understood as an essential precursor to the internal reorganization that is necessary for the amelioration of psychological distress. PCT suggests a more considered and widespread use of exposure in psychotherapy as a way of improving both the efficiency and the effectiveness of the treatments offered. Despite the proliferation of psychotherapies in recent years there has not been a commensurate growth in our understanding of the effective ingredients of treatment. It seems unlikely that there are multitudes of different mechanisms and processes through which psychological distress is resolved and that each of these different psychotherapies utilises a distinct item from the collection. In fact, the spawning of hundreds of different psychotherapies is perhaps the most telling sign that there may still be much work to do before the important principles of treatment are described and fundamental mechanisms of change are identified. Could a technique as mundane as exposure hold the key to effective psychotherapy? For psychotherapy, is there just one road leading to Rome but a plethora of ways to travel that road?


Reproductive coercion: an under-recognized challenge for primary care patients.
Reproductive coercion impacts many women of reproductive age. We sought to explore how reproductive coercion, including pregnancy coercion and birth control sabotage, impacts women in a primary care population. We administered a survey to women accessing care at a family medicine clinic in the Bronx, NY. Reproductive coercion was defined as a positive response to at least one of five questions adapted from previous studies. We assessed the association of reproductive and demographic characteristics with a lifetime history of reproductive coercion. At least one form of reproductive coercion was reported by 24% of the 97 respondents. Current lack of personal safety and a history of transactional sex for money or a place to stay were significantly associated with having experienced reproductive coercion (all P ≤ 0.02). Reproductive coercion was common among women of reproductive age at this urban family medicine clinic in an underserved community, and was associated with other forms of control and violence. Clinicians are advised to discuss birth control sabotage and pregnancy coercion with their patients.


GSK3 and β-catenin determines functional expression of sodium channels at the axon initial segment.
Neuronal action potentials are generated through voltage-gated sodium channels, which are tethered by ankyrinG at the membrane of the axon initial segment (AIS). Despite the importance of the AIS in the control of neuronal excitability, the cellular and molecular mechanisms regulating sodium channel expression at the AIS remain elusive. Our results show that GSK3α/β and β-catenin phosphorylated by GSK3 (S33/37/T41) are localized at the AIS and are new components of this essential neuronal domain. Pharmacological inhibition of GSK3 or β-catenin knockdown with shRNAs decreased the levels of phosphorylated-β-catenin, ankyrinG, and voltage-gated sodium channels at the AIS, both "in vitro" and "in vivo", therefore diminishing neuronal excitability as evaluated via sodium current amplitude and action potential number. Thus, our results suggest a mechanism for the modulation of neuronal excitability through the control of sodium channel density by GSK3 and β-catenin at the AIS.


The Spin-Partitioned Total-Position Spread Tensor: An Application To Diatomic Molecules.
The spin partition (SP) of the total-position spread (TPS) tensor is applied to the case of a few light diatomic molecules at full configuration interaction (FCI) level. It appears that the SP-TPS tensor gives informations that are complementary with respect to the corresponding spin-summed (SS) quantity. The spin-summed total position-spread tensor (SS-TPS, Λ) is defined as the second moment cumulant of the total position operator, and the SP-TPS is its partition in equal (Λαα+ββ) and different spin (Λαβ+βα) contributions. Then, the SS-TPS allows description of the molecule charge mobility, while the SP-TPS allows description of the spin delocalization. The most relevant Cartesian-component for both tensors (SS-TPS and SP-TPS) is the component along the chemical bond (Λ(∥)), and it was found that its behavior was related to the type of interaction involved. For covalent bonds the SP-TPS has a squared growth when the bond is stretched, while for ionic bonds there exists a faster-than-linear growth after the avoided-crossing between the covalent and the ionic states. Other exotic bonds, like He2 and Be2, were also considered, and a particular spin delocalization was able to describe the different character of the two weakly bonded molecules, and specially the multireference character of the wave function along the dissociative potential energy curve.


Interleukin-33-induced expression of PIBF1 by decidual B cells protects against preterm labor.
Preterm birth (PTB) is a leading cause of neonatal death worldwide. Intrauterine and systemic infection and inflammation cause 30-40% of spontaneous preterm labor (PTL), which precedes PTB. Although antibody production is a major immune defense mechanism against infection, and B cell dysfunction has been implicated in pregnancy complications associated with PTL, the functions of B cells in pregnancy are not well known. We found that choriodecidua of women undergoing spontaneous PTL harbored functionally altered B cell populations. B cell-deficient mice were markedly more susceptible than wild-type (WT) mice to PTL after inflammation, but B cells conferred interleukin (IL)-10-independent protection against PTL. B cell deficiency in mice resulted in a lower uterine level of active progesterone-induced blocking factor 1 (PIBF1), and therapeutic administration of PIBF1 mitigated PTL and uterine inflammation in B cell-deficient mice. B cells are a significant producer of PIBF1 in human choriodecidua and mouse uterus in late gestation. PIBF1 expression by B cells is induced by the mucosal alarmin IL-33 (ref. 9). Human PTL was associated with diminished expression of the α-chain of IL-33 receptor on choriodecidual B cells and a lower level of active PIBF1 in late gestation choriodecidua. These results define a vital regulatory cascade involving IL-33, decidual B cells and PIBF1 in safeguarding term pregnancy and suggest new therapeutic approaches based on IL-33 and PIBF1 to prevent human PTL.


Neuronal nitric oxide synthase and human vascular regulation.
Vascular blood flow and its distribution among different vascular beds are regulated by changes in microvascular tone. Nitric oxide (NO) plays a key role in the local paracrine regulation of vessel tone both under resting conditions and when blood flow increases in response to agonist stimulation or increased shear stress. The conventional notion that endothelial NO synthase (eNOS)-derived NO is largely responsible for both effects has been challenged by first-in-human studies with a selective inhibitor of neuronal NOS (nNOS), S-methyl-l-thiocitrulline (SMTC). These studies reveal that SMTC causes a reduction in basal blood flow in the normal human forearm and coronary circulations (that is reversed by l-arginine), without affecting the eNOS-mediated vasodilatation elicited by acetylcholine, substance P, or increased shear stress. S-methyl-l-thiocitrulline also inhibits mental stress-induced vasodilatation. These results are consistent with a significant body of experimental studies suggesting that nNOS plays an important role in the local regulation of vessel tone in other species, independent of the effects of nNOS-derived NO in the central nervous system. These emerging data suggest that eNOS and nNOS have distinct roles in the physiologic local regulation of human microvascular tone in vivo and pave the way for further detailed investigation of the relative contribution of nNOS and eNOS in vascular regulation in human disease.


Consistency of blood pressure differences between the left and right arms.
It is unclear to what extent interarm blood pressure (BP) differences are reproducible vs the result of random error. The present study was designed to resolve this issue. We enrolled 147 consecutive patients from a hypertension clinic. Three sets of 3 BP readings were recorded, first using 2 oscillometric devices simultaneously in the 2 arms (set 1); next, 3 readings were taken sequentially for each arm using a standard mercury sphygmomanometer (set 2); finally, the readings as performed for set 1 were repeated (set 3). The protocol was repeated at a second visit for 91 patients. Large interarm systolic BP differences were consistently seen in 2 patients with obstructive arterial disease. In the remaining patients, the systolic BP and the diastolic BP, respectively, were slightly higher in the right arm than in the left arm by 2 to 3 mm Hg and by 1 mm Hg for all 3 sets (P<.01 for all). For the systolic BP and the diastolic BP, respectively, the numbers of patients who had a mean interarm difference of more than 5 mm Hg were 11 (7.5%) and 4 (2.7%) across all 3 sets of readings. Among patients who repeated the test, none had a consistent interarm BP difference of more than 5 mm Hg across the 2 visits. The interarm BP difference was consistent only when obstructive arterial disease was present. Although BP in the right arm tended to be higher than in the left arm, clinically meaningful interarm differences were not reproducible in the absence of obstructive arterial disease and are attributable to random variation.


Systematic review of evidence relating to the treatment of immune-mediated hemolytic anemia in dogs.
Despite being the most prevalent autoimmune disease of dogs, there is considerable variation between individuals and institutions in the treatment regimens that are employed for the management of immune-mediated hemolytic anemia. The aim of this review was to evaluate evidence relating to the treatment of the disease systematically and to use this evidence to draw conclusions that are applicable in wider veterinary practice. Search tools were employed to identify relevant articles and these were assessed according to stated criteria. The overall quality of published evidence was poor, with many articles failing to provide details of the enrollment, treatment, monitoring, and assessment stages of the study process. In view of this, firm conclusions cannot be drawn regarding the treatment of this disease and further research of a higher quality is required.


Proresolving lipid mediators resolvin D1, resolvin D2, and maresin 1 are critical in modulating T cell responses.
Resolution of inflammation is a finely regulated process mediated by specialized proresolving lipid mediators (SPMs), including docosahexaenoic acid (DHA)-derived resolvins and maresins. The immunomodulatory role of SPMs in adaptive immune cells is of interest. We report that D-series resolvins (resolvin D1 and resolvin D2) and maresin 1 modulate adaptive immune responses in human peripheral blood lymphocytes. These lipid mediators reduce cytokine production by activated CD8(+) T cells and CD4(+) T helper 1 (TH1) and TH17 cells but do not modulate T cell inhibitory receptors or abrogate their capacity to proliferate. Moreover, these SPMs prevented naïve CD4(+) T cell differentiation into TH1 and TH17 by down-regulating their signature transcription factors, T-bet and Rorc, in a mechanism mediated by the GPR32 and ALX/FPR2 receptors; they concomitantly enhanced de novo generation and function of Foxp3(+) regulatory T (Treg) cells via the GPR32 receptor. These results were also supported in vivo in a mouse deficient for DHA synthesis (Elovl2(-/-)) that showed an increase in TH1/TH17 cells and a decrease in Treg cells compared to wild-type mice. Additionally, either DHA supplementation in Elovl2(-/-) mice or in vivo administration of resolvin D1 significantly reduced cytokine production upon specific stimulation of T cells. These findings demonstrate actions of specific SPMs on adaptive immunity and provide a new avenue for SPM-based approaches to modulate chronic inflammation.


Influenza virus receptor specificity and cell tropism in mouse and human airway epithelial cells.
Recent human infections caused by the highly pathogenic avian influenza virus H5N1 strains emphasize an urgent need for assessment of factors that allow viral transmission, replication, and intra-airway spread. Important determinants for virus infection are epithelial cell receptors identified as glycans terminated by an alpha2,3-linked sialic acid (SA) that preferentially bind avian strains and glycans terminated by an alpha2,6-linked SA that bind human strains. The mouse is often used as a model for study of influenza viruses, including recent avian strains; however, the selectivity for infection of specific respiratory cell populations is not well described, and any relationship between receptors in the mouse and human lungs is incompletely understood. Here, using in vitro human and mouse airway epithelial cell models and in vivo mouse infection, we found that the alpha2,3-linked SA receptor was expressed in ciliated airway and type II alveolar epithelial cells and was targeted for cell-specific infection in both species. The alpha2,6-linked SA receptor was not expressed in the mouse, a factor that may contribute to the inability of some human strains to efficiently infect the mouse lung. In human airway epithelial cells, alpha2,6-linked SA was expressed and functional in both ciliated and goblet cells, providing expanded cellular tropism. Differences in receptor and cell-specific expression in these species suggest that differentiated human airway epithelial cell cultures may be superior for evaluation of some human strains, while the mouse can provide a model for studying avian strains that preferentially bind only the alpha2,3-linked SA receptor.


Simple inversion technique to obtain cloud droplet size parameters using solar aureole data.
A simple inversion technique in the single scattering regime has been developed to deduce cloud droplet size parameters by using the measurement of the radiance of near-forward scattered solar radiation as a function of angle. Compared with the numerical inversion technique that uses exact Mie scattering calculations, the new technique is much less time-consuming and hence should be usable in an on-line real time analysis. To test the effectiveness of the new technique, we use the results of polydispersed cloud size distribution calculated by Deirmendjian to retrieve the model size parameters. The agreement is excellent. We also generalize the theory to include the broadband source. A typical experimental example is given. Its comparison with time-consuming Mie scattering inversion technique again shows excellent agreement.


A new and simple solid-phase extraction method for LC determination of pyronaridine in human plasma.
A new approach using a simple solid-phase extraction technique has been developed for the determination of pyronaridine (PND), an antimalarial drug, in human plasma. After extraction with C18 solid-phase sorbent, PND was analyzed using a reverse phase chromatographic method with fluorescence detection (at lambda(ex)=267 nm and lambda(em)=443 nm). The mean extraction recovery for PND was 95.2%. The coefficient of variation for intra-assay precision, inter-assay precision and accuracy was less than 10%. The quantification limit with fluorescence detection was 0.010 microg/mL plasma. The method described herein has several advantages over other published methods since it is easy to perform and rapid. It also permits reducing both, solvent use and sample preparation time. The method has been used successfully to assay plasma samples from clinical pharmacokinetic studies.


Training specificity of hurdle vs. countermovement jump training.
The objective of this study was to compare bilateral and unilateral hurdle jumps with traditional countermovement jumps (CMJs). Thirteen athletes were tested during continuous forward bilateral and unilateral hurdle jumps and single CMJ. Countermovement jump height was used to establish the hurdle height. Subjects jumped forward over 4 hurdles with the force plate positioned after the second hurdle to measure vertical ground reaction force (VGRF), contact time (CT), and rate of force development (RFD). For bilateral jumps, hurdle height was established at maximal (100%) CMJ height and at 120, 140, and 160% of the CMJ height. The athletes were instructed to jump as fast as possible to mimic a training session drill. For unilateral jumps, hurdle height was set at 70, 80, and 90% of the CMJ height. Bilateral 160% jumps showed a significantly longer CT than bilateral 100, 120, and 140% jumps. The bilateral 100, 120, and 140% jumps had significantly shorter CT than the unilateral jumps and CMJ. The VGRF during bilateral jumps was higher than unilateral jumps and CMJ. Bilateral 160% jump RFD was significantly higher than CMJ and unilateral jumps but significantly lower than the other bilateral jumps. In conclusion, the characteristics of the bilateral jumps were substantially different from those of the CMJ and unilateral hurdle jumps. As bilateral hurdle jumps with a height between 100 and 140% of the CMJ provide similar CTs and VGRF as many reported sprint or jump actions, they may be considered a more training-specific power training drill than the CMJ.


Double-negative T cells, activated by xenoantigen, lyse autologous B and T cells using a perforin/granzyme-dependent, Fas-Fas ligand-independent pathway.
The ability to control the response of B cells is of particular interest in xenotransplantation as Ab-mediated hyperacute and acute xenograft rejection are major obstacles in achieving long-term graft survival. Regulatory T cells have been proven to play a very important role in the regulation of immune responses to self or non-self Ags. Previous studies have shown that TCRalphabeta+CD3+CD4-CD8- (double-negative (DN)) T cells possess an immune regulatory function, capable of controlling antidonor T cell responses in allo- and xenotransplantation through Fas-Fas ligand interaction. In this study, we investigated the possibility that xenoreactive DNT cells suppress B cells. We found that DNT cells generated from wild-type C57BL/6 mice expressed B220 and CD25 after rat Ag stimulation. These xenoreactive B220+CD25+ DNT cells lysed activated, but not naive, B and T cells. This killing, which took place through cell-cell contact, required participation of adhesion molecules. Our results indicate that Fas ligand, TGF-beta, TNF-alpha, and TCR-MHC recognition was not involved in DNT cell-mediated syngenic cell killing, but instead this killing was mediated by perforin and granzymes. The xenoreactive DNT cells expressed high levels of granzymes in comparison to allo- or xenoreactive CD8+ T cells. Adoptive transfer of DNT cells in combination with early immune suppression by immunosuppressive analog of 15-deoxyspergualin, LF15-0195, significantly prolonged rat heart graft survival to 62.1 +/- 13.9 days in mice recipients. In conclusion, this study suggests that xenoreactive DNT cells can control B and T cell responses in perforin/granzyme-dependent mechanisms. DNT cells may be valuable in controlling B and T cell responses in xenotransplantation.


Social losses predict a faster onset and greater severity of obsessive-compulsive disorder.
While stressful life events increase the risk of developing a range of psychiatric disorders, including obsessive-compulsive disorder (OCD), their ability to precipitate specific obsessive-compulsive symptoms' dimensions is unknown. Here we aimed to evaluate the potential role of three different types of stressful life events, herein termed losses (death of a loved one, termination of a romantic relationship and severe illness) in predicting the speed of progression from subclinical to clinical OCD and the severity of specific OCD dimensions in a large multicentre OCD sample. Nine hundred and fifty-four OCD outpatients from the Brazilian OCD Research Consortium were included in this study. Several semi-structured and structured instruments were used, including the Structured Clinical Interview for DSM-IV Axis I Disorders, the Yale-Brown Obsessive-Compulsive Scale, the Dimensional Yale-Brown Obsessive-Compulsive Scale, the Brown Assessment of Beliefs Scale, the Beck Depression Inventory, the Beck Anxiety Inventory and the Yale OCD Natural History Questionnaire. Regression models investigated the interaction between types of loss and gender to predict speed of progression from subclinical obsessive-compulsive symptoms to OCD, and the severity of five symptom dimensions. While termination of a relationship was associated with a faster speed of progression from subthreshold to clinical OCD, the death of a loved one was associated with increased severity of hoarding symptoms. There was also an interaction between gender and experiences of death, which predicted a faster speed of progression to OCD in males. Stressful life events have the ability to accelerate the progression from subclinical to clinical OCD, as well as impact the severity of specific OCD dimensions. Gender also plays a role in both the progression and severity of symptoms. These findings suggest that stressful life events may represent a marker to identify individuals at risk of progressing to clinical OCD.


Vascular responses to prostaglandin F 2 alpha in isolated cat lungs.
Changes in perfusion pressure in response to graded doses of prostaglandin F 2 alpha (PGF 2 alpha) were measured during both forward and retrograde perfusion of isolated cat lungs perfused at a constant flow rate. During forward perfusion, PGF 2 alpha produced a dose-dependent increase in pulmonary artery pressure and a decrease in lung fluid volume. During retrograde perfusion, PGF 2 alpha also produced a dose-dependent increase in perfusion pressure; however, the dose required was fivefold greater than that needed to produce an identical change in pressure during forward perfusion. In addition, during retrograde perfusion, the lung fluid volume increased in response to PGF 2 alpha. These results suggest that the major site of activity of PGF 2 alpha is on the arterial side of the pulmonary vascular bed and that inactivation of PGF 2 alpha by the lung occurs primarily distal to this arterial site of vasomotion. The changes in perfusion pressure in response to PGF 2 alpha were markedly dependent on pH and oxygen tension (Po-2), being abolished by severe alkalosis and potentiated by both acidosis and hypoxia. In contrast, neither serotonin nor norepinephrine exhibited such a pH or Po-2 dependency. Since the ratio of the forward response to the retrograde response was not decreased by alterations of pH or Po-2, their influence on the responses appears to be through interaction at the site of vascular activity rather than through alteration of the rate of prostaglandin inactivation.


Emission of polycyclic aromatic hydrocarbons and their carcinogenic potencies from cooking sources to the urban atmosphere.
Traffic has long been recognized as the major contributor to polycyclic aromatic hydrocarbon (PAH) concentrations. However, this does not consider the contribution of cooking sources of PAHs. This study set out, first, to assess the characteristics of PAHs and their corresponding benzo[a]pyrene equivalent (B[a]Peq) emissions from cooking sources to the urban atmosphere. To illustrate the importance of cooking sources, PAH emissions from traffic sources were then calculated and compared. The entire study was conducted on a city located in southern Taiwan. PAH samples were collected from the exhaust stacks of four types of restaurant: Chinese, Western, fast food, and Japanese. For total PAHs, results show that the fractions of gaseous PAHs (range, 75.9-89.9%) were consistently higher than the fractions of particulate PAHs (range, 10.1-24.1%) in emissions from the four types of restaurant. But for total B[a]Peq, we found that the contributions of gaseous PAHs (range, 15.7-21.9%) were consistently lower than the contributions of particulate PAHs (range, 78.1-84.3%). For emission rates of both total PAHs and total B[a]Peq, a consistent trend was found for the four types of restaurant: Chinese (2,038 and 154 kg/year, respectively) > Western (258 and 20.4 kg/year, respectively) > fast food (31.4 and 0.104 kg/year, respectively) > Japanese (5.11 and 0.014 kg/year, respectively). By directly adapting the emission data obtained from Chinese restaurants, we found that emission rates on total PAHs and total B[a]Peq for home kitchen sources were 6,639 and 501 kg/year, respectively. By combining both restaurant sources and home kitchen sources, this study yielded emission rates of total PAHs and total B[a]Peq from cooking sources of the studied city of 8,973 and 675 kg/year, respectively. Compared with PAH emissions from traffic sources in the same city, we found that although the emission rates of total PAHs for cooking sources were significantly less than those for traffic sources (13,500 kg/year), the emission rates of total B[a]Peq for cooking sources were much higher than those for traffic sources (61.4 kg/year). The above results clearly indicate that although cooking sources are less important than traffic sources in contributing to total PAH emissions, PAH emissions from cooking sources might cause much more serious problems than traffic sources, from the perspective of carcinogenic potency.


Development and validation of a novel reporter assay for human papillomavirus type 16 late gene expression.
To facilitate the investigations of HPV-16 late gene expression HPV-16 reporter plasmids were generated using previously described sub-genomic HPV-16 plasmids, named pBEL and pBELM, that, similar to the full viral genome, produce primarily HPV-16 early mRNAs and very little, if any, late mRNAs in cervical cancer cells. The HPV-16 late L1 gene was replaced by the chloramphenicol acetyltransferase (CAT) reporter gene, or green fluorescent protein (GFP), preceded by the poliovirus internal ribosome entry site (IRES). Results show that the reporter genes mimic the expression of L1 from these plasmids. For example, overexpression of adenovirus E4orf4 protein (E4orf4), polypyrimidine tract binding protein (PTB), arginine/serine-rich SRp30c protein (SRp30c) or alternative splicing factor/splicing factor 2 (ASF/SF2) induced an increased expression of CAT or GFP. Stable cell lines with reporter plasmids pBELCAT and pBELMCAT were also generated. An induction of CAT was observed in HPV-16 reporter cell lines in the presence of the small molecule phorbol 12-myristate 13-acetate (TPA). Further experiments identified the TPA-inducible, hnRNP A2/B1 protein as a regulator of HPV-16 late gene expression. In conclusion, the HPV-16 reporter plasmids and reporter cell lines described herein can be used to identify small molecules and cellular factors that regulate HPV-16 gene expression.


Surface Plasmon Resonance Analysis of PfEMP1 Interaction with Receptors.
A detailed understanding of the interaction between the highly variant Plasmodium falciparum erythrocyte membrane proteins 1 (PfEMP1) and their human binding partners is essential to explain their roles in disease development in malaria, as well as to understand how antibodies can inhibit these interactions and how the parasite manages to evade such an immune response. This chapter focuses on using surface plasmon resonance (SPR) as a reproducible, high-throughput method to quantitatively characterize these interactions. We describe how to utilize protein A or A/G and streptavidin for protein immobilization on SPR sensor chips and provide instructions on how to biotinylate proteins for this purpose and how to use SPR for binding competition assays. Since these experiments rely on recombinant proteins, we also present a method to verify their structural integrity using circular dichroism spectroscopy.


High spatial frequency periodic structures induced on metal surface by femtosecond laser pulses.
The high spatial frequency periodic structures induced on metal surface by femtosecond laser pulses was investigated experimentally and numerically. It is suggested that the redistribution of the electric field on metal surface caused by the initially formed low spatial frequency periodic structures plays a crucial role in the creation of high spatial frequency periodic structures. The field intensity which is initially localized in the grooves becomes concentrated on the ridges in between the grooves when the depth of the grooves exceeds a critical value, leading to the ablation of the ridges in between the grooves and the formation of high spatial frequency periodic structures. The proposed formation process is supported by both the numerical simulations based on the finite-difference time-domain technique and the experimental results obtained on some metals such as stainless steel and nickel.


Plasma Steroid Metabolome Profiling for Diagnosis and Subtyping Patients with Cushing Syndrome.
Diagnosis of Cushing syndrome requires a multistep process that includes verification of hypercortisolism followed by identification of the cause of adrenocortical hyperfunction. This study assessed whether pituitary, ectopic, and adrenal subtypes of Cushing syndrome were characterized by distinct plasma steroid profiles that might assist diagnosis. In this retrospective cross-sectional study, mass spectrometric measurements of a panel of 15 plasma steroids were applied to 222 patient samples tested for Cushing syndrome. Disease was excluded in 138 and confirmed in 51 patients with pituitary Cushing syndrome, 12 with ectopic adrenocorticotropin secretion, and 21 with adrenal disease. Another 277 age- and sex-matched hypertensive and normotensive volunteers were included for comparison. Compared with patients without disease, the largest increases in plasma steroids among patients with Cushing syndrome were observed for 11-deoxycortisol (289%), 21-deoxycortisol (150%), 11-deoxycorticosterone (133%), corticosterone (124%), and cortisol (122%). Patients with ectopic disease showed the most prominent increases, but there was considerable variation for other steroids according to subtype. Patients with adrenal disease had the lowest concentrations of androgens, whereas those with ectopic and pituitary disease showed the lowest concentrations of aldosterone. Plasma 18-oxocortisol was particularly low in ectopic disease. With the use of 10 selected steroids, subjects with and without different Cushing syndrome subtypes could be discriminated nearly as closely as with the use of salivary and urinary free cortisol, dexamethasone-suppressed cortisol, and plasma adrenocorticotropin (9.5% vs 5.8% misclassification). Patients with different subtypes of Cushing syndrome show distinctive plasma steroid profiles that may offer a supplementary single-test alternative for screening purposes.


Using association rules mining to characterize loss of control eating in childhood.
Childhood loss of control (LOC)-eating, the perceived inability to stop or control eating, is associated with increased risk for binge-eating disorder and obesity. However, the correlates of LOC-eating in childhood remain unclear. A secondary analysis of 177, 7-12-year-old children from five laboratory feeding studies was performed to investigate potential family (e.g., frequency of meals together, feeding practices), parental (e.g., education, weight status), and child (e.g., weight status, appetite traits) correlates of LOC-eating. Association rules mining (ARM1), a data-driven approach, was used to examine all characteristics that were common across studies to identify which were associated with LOC-eating. Results showed LOC-eating was characterized by a combination of child appetitive behaviors and parental feeding practices. In particular, LOC-eating was associated with low parental pressure to eat in combination with a high propensity to want to eat all the time and frequent refusal or dislike of novel foods. This pattern of both food approach (i.e., wanting to eat all the time) and avoidant behaviors (i.e., food fussiness) highlights the need for more research to characterize the complex patterns of appetitive traits associated with LOC-eating. In contrast, the absence of LOC-eating was associated with a low propensity to want to eat all the time, greater family income, and infrequent emotional overeating. Therefore, propensity to want to eat all the time, a single question from the Children's Eating Behavior Questionnaire, characterized both the presence and absence of LOC-eating, highlighting the need for more research to determine if this question captures clinically relevant individual differences. Future studies addressing these questions will advance our understanding of pediatric LOC-eating and may lead to interventions to reduce risk for more severe eating disorder symptomology.