Discovery of insect and human dengue virus host factors.
Dengue fever is the most frequent arthropod-borne viral disease of humans, with almost half of the world's population at risk of infection. The high prevalence, lack of an effective vaccine, and absence of specific treatment conspire to make dengue fever a global public health threat. Given their compact genomes, dengue viruses (DENV-1-4) and other flaviviruses probably require an extensive number of host factors; however, only a limited number of human, and an even smaller number of insect host factors, have been identified. Here we identify insect host factors required for DENV-2 propagation, by carrying out a genome-wide RNA interference screen in Drosophila melanogaster cells using a well-established 22,632 double-stranded RNA library. This screen identified 116 candidate dengue virus host factors (DVHFs). Although some were previously associated with flaviviruses (for example, V-ATPases and alpha-glucosidases), most of the DVHFs were newly implicated in dengue virus propagation. The dipteran DVHFs had 82 readily recognizable human homologues and, using a targeted short-interfering-RNA screen, we showed that 42 of these are human DVHFs. This indicates notable conservation of required factors between dipteran and human hosts. This work suggests new approaches to control infection in the insect vector and the mammalian host.


Genetic analysis of salinity tolerance in rice (Oryza sativa L.).
The genetics of salinity tolerance in rice was investigated by a nine-parent complete diallel including reciprocals. Test materials involved susceptible (IR28, IR29, and MI-48), moderately tolerant (IR4595-4-1-13, IR9884-54-3-1E-P1, and IR10206-29-2-1), and tolerant ("Nona Bokra", "Pokkali", and SR26B) parents. Twoweek-old seedlings were grown in a salinized (EC = 12 dS/m) culture solution for 19 days under controlled conditions in the IRRI phytotron. Typical characteristics of salinity tolerance in rice were found to be Na(+) exclusion and an increased absorption of K(+) to maintain a good Na-K balance in the shoot. Genetic component analysis (GCA) revealed that a low Na-K ratio is governed by both additive and dominance gene effects. The trait exhibited overdominance, and two groups of genes were detected. Environmental effects were large, and the heritability of the trait was low. Our findings suggest that when breeding for salt tolerance, selection must be done in a later generation and under controlled conditions in order to minimize environmental effects. Modified bulk and single-seed descent would be the suitable breeding methods. Combining ability analysis revealed that both GCA and specific combining ability (SCA) effects were important in the genetics of salt tolerance. Moderately tolerant parents - e.g., IR4595-4-1-13 and IR9884-54-3-1E-P1 - were the best general combiners. Most of the best combinations had susceptible parents crossed either to moderate or tolerant parents. The presence of reciprocal effects among crosses necessitates the use of susceptible parents as males in hybridization programs. Large heterotic effects suggest the potential of hybrid rice for salt-affected lands.


Bt-maize (MON810) and non-GM soybean meal in diets for Atlantic salmon (Salmo salar L.) juveniles--impact on survival, growth performance, development, digestive function, and transcriptional expression of intestinal immune and stress responses.
Responses in Atlantic salmon (Salmo salar L.) juveniles (fry) fed diets containing genetically modified maize (Bt-maize, MON810) expressing Cry1Ab protein from first-feeding were investigated during a 99-day feeding trial. Four experimental diets were made; each diet contained ∼20% maize, either Bt-maize or its near-isogenic maternal line (non-GM maize). One pair was fishmeal-based while the other pair included standard (extracted) soybean meal (SBM; 16.7% inclusion level), with the intention of investigating responses to the maize varieties in healthy fish as well as in immunologically challenged fish with SBM-induced distal intestinal inflammation, respectively. Three replicate tanks of fry (0.17±0.01 g; initial mean weight ± SEM) were fed one of the four diets and samples were taken on days 15, 36, 48 and 99. Survival, growth performance, whole body composition, digestive function, morphology of intestine, liver and skeleton, and mRNA expression of some immune and stress response parameters in the distal intestine were evaluated. After 99 days of feeding, survival was enhanced and the intended SBM-induced inflammatory response in the distal intestine of the two groups of SBM-fed fish was absent, indicating that the juvenile salmon were tolerant to SBM. Mortality, growth performance and body composition were similar in fish fed the two maize varieties. The Bt-maize fed fish, however, displayed minor but significantly decreased digestive enzyme activities of leucine aminopeptidase and maltase, as well as decreased concentration of gut bile salts, but significantly increased amylase activity at some sampling points. Histomorphological, radiographic and mRNA expression evaluations did not reveal any biologically relevant effects of Bt-maize in the gastrointestinal tract, liver or skeleton. The results suggest that the Cry1Ab protein or other compositional differences in GM Bt-maize may cause minor alterations in intestinal responses in juvenile salmon, but without affecting overall survival, growth performance, development or health.


Controlling Geometry and Flow Through Bacterial Bridges on Patterned Lubricant-Infused Surfaces (pLIS).
Spatial control of bacteria and biofilms on surfaces is necessary to understand the biofilm formation and the social interactions between bacterial communities, which could provide useful hints to study the biofilm-involved diseases. Here patterned lubricant-infused surfaces (pLIS) are utilized to fabricate connective structures named "bacterial bridges" between bacterial colonies of Pseudomonas aeruginosa by a simple dewetting method. It is demonstrated that the bacteria attached to hydrophilic areas and bacteria precipitated on lubricant infused borders both contribute to the formation of bacterial bridges. The geometry and distribution of bridges can be controlled using predesigned superhydrophobic-hydrophilic patterns. It is demonstrated that bacterial bridges connecting bacteria colonies act as bio-microfluidic channels and can transport liquids, nutrients, and antibacterial substances between neighboring bacteria clusters. Thus, bacterial bridges can be used to study formation, spreading, and development of bacterial colonies, and communication within and between isolated biofilms.


Characterization of an inducible promoter in different DNA copy number conditions.
The bottom-up programming of living organisms to implement novel user-defined biological capabilities is one of the main goals of synthetic biology. Currently, a predominant problem connected with the construction of even simple synthetic biological systems is the unpredictability of the genetic circuitry when assembled and incorporated in living cells. Copy number, transcriptional/translational demand and toxicity of the DNA-encoded functions are some of the major factors which may lead to cell overburdening and thus to nonlinear effects on system output. It is important to disclose the linearity working boundaries of engineered biological systems when dealing with such phenomena. The output of an N-3-oxohexanoyl-L-homoserine lactone (HSL)-inducible RFP-expressing device was studied in Escherichia coli in different copy number contexts, ranging from 1 copy per cell (integrated in the genome) to hundreds (via multicopy plasmids). The system is composed by a luxR constitutive expression cassette and a RFP gene regulated by the luxI promoter, which is activated by the HSL-LuxR complex. System output, in terms of promoter activity as a function of HSL concentration, was assessed relative to the one of a reference promoter in identical conditions by using the Relative Promoter Units (RPU) approach. Nonlinear effects were observed in the maximum activity, which is identical in single and low copy conditions, while it decreases for higher copy number conditions. In order to properly compare the luxI promoter strength among all the conditions, a mathematical modeling approach was used to relate the promoter activity to the estimated HSL-LuxR complex concentration, which is the actual activator of transcription. During model fitting, a correlation between the copy number and the dissociation constant of HSL-LuxR complex and luxI promoter was observed. Even in a simple inducible system, nonlinear effects are observed and non-trivial data processing is necessary to fully characterize its operation. The in-depth analysis of model systems like this can contribute to the advances in the synthetic biology field, since increasing the knowledge about linearity and working boundaries of biological phenomena could lead to a more rational design of artificial systems, also through mathematical models, which, for example, have been used here to study hard-to-predict interactions.


A Collaborative Multidisciplinary Approach to the Management of Coronavirus Disease 2019 in the Hospital Setting.
The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), which presents an unprecedented challenge to medical providers worldwide. Although most SARS-CoV-2-infected individuals manifest with a self-limited mild disease that resolves with supportive care in the outpatient setting, patients with moderate to severe COVID-19 will require a multidisciplinary collaborative management approach for optimal care in the hospital setting. Laboratory and radiologic studies provide critical information on disease severity, management options, and overall prognosis. Medical management is mostly supportive with antipyretics, hydration, oxygen supplementation, and other measures as dictated by clinical need. Among its medical complications is a characteristic proinflammatory cytokine storm often associated with end-organ dysfunction, including respiratory failure, liver and renal insufficiency, cardiac injury, and coagulopathy. Specific recommendations for the management of these medical complications are discussed. Despite the issuance of emergency use authorization for remdesivir, there are still no proven effective antiviral and immunomodulatory therapies, and their use in COVID-19 management should be guided by clinical trial protocols or treatment registries. The medical care of patients with COVID-19 extends beyond their hospitalization. Postdischarge follow-up and monitoring should be performed, preferably using telemedicine, until the patients have fully recovered from their illness and are released from home quarantine protocols.


Patterns of gene action in plant development revealed by enhancer trap and gene trap transposable elements.
The crucifer Arabidopsis thaliana has been used widely as a model organism for the study of plant development. We describe here the development of an efficient insertional mutagenesis system in Arabidopsis that permits identification of genes by their patterns of expression during development. Transposable elements of the Ac/Ds system carrying the GUS reporter gene have been designed to act as enhancer traps or gene traps. A novel selection scheme maximizes recovery of unlinked transposition events. In this study 491 plants carrying independent transposon insertions were generated and screened for expression patterns. One-half of the enhancer trap insertions and one-quarter of the gene trap insertions displayed GUS expression in seedlings or flowers, including expression patterns specific to organs, tissues, cell types, or developmental stages. The patterns identify genes that act during organogenesis, pattern formation, or cell differentiation. Transposon insertion lines with specific GUS expression patterns provide valuable markers for studies of Arabidopsis development and identify new cell types or subtypes in plants. The diversity of gene expression patterns generated suggests that the identification and cloning of Arabidopsis genes expressed in any developmental process is feasible using this system.


Diagnostic and predictive accuracy of placental growth factor in suspected pre-eclampsia.
The diagnosis of preeclampsia can be difficult with considerable uncertainty when the presenting features are borderline. Moreover underlying medical conditions may obscure or simulate the onset of pre-eclampsia. There is a clinical need to be able to diagnose, more accurately, those forms of the disease that are likely to progress to an adverse fetal or maternal outcome. Maternal circulating placental growth factor (PIGF) concentrations are decreased in these women, in correlation with disease severity and gestational age of onset. In this study its diagnostic performance was evaluated with an accurate, point of care test. The PELICAN study was a prospective observational cohort study, in seven maternity units in the British Isles. 625 women presenting between 20+0 and 40+6 weeks gestation with suspected preeclampsia were recruited, excluding those already diagnosed with preeclampsia. PIGF was measured (Alere Triage® PIGF test) in the enrolment sample after all participants had delivered so that clinical decisions were not affected by its results. ISSHP definitions of hypertensive disease were assigned, blinded to PIGF values. Analysis was conducted to assess diagnostic accuracy of the enrolment PIGF level for placental dysfunction requiring delivery within 14days. In women who presented before 35 week's gestation the PIGF test predicted placental dysfunction, that required deliverywithin the next 14days, with the following characteristics: sensitivity 0.95 (0.86-0.98); specificity 0.56 [0.49-0.63); NPV 0.97 (0.92-0.99); PPV0.42 (0.35-0.51). PIGF is a strong predictor of time-to-delivery and may be an independent and complementary tool to assist in diagnosis and risk stratification of pregnancies complicated by placental dysfunction. These valuable attributes are to be tested in prospecting randomised double-blind controlled trial.


Topographic features of nano-pores within the osteochondral interface and their effects on transport properties -a 3D imaging and modeling study.
Recent insights suggest that the osteochondral interface plays a central role in maintaining healthy articulating joints. Uncovering the underlying transport mechanisms is key to the understanding of the cross-talk between articular cartilage and subchondral bone. Here, we describe the mechanisms that facilitate transport at the osteochondral interface. Using scanning electron microscopy (SEM), we found a continuous transition of mineralization architecture from the non-calcified cartilage towards the calcified cartilage. This refurbishes the classical picture of the so-called tidemark; a well-defined discontinuity at the osteochondral interface. Using focused-ion-beam SEM (FIB-SEM) on one osteochondral plug derived from a human cadaveric knee, we elucidated that the pore structure gradually varies from the calcified cartilage towards the subchondral bone plate. We identified nano-pores with radius of 10.71 ± 6.45 nm in calcified cartilage to 39.1 ± 26.17 nm in the subchondral bone plate. The extracted pore sizes were used to construct 3D pore-scale numerical models to explore the effect of pore sizes and connectivity among different pores. Results indicated that connectivity of nano-pores in calcified cartilage is highly compromised compared to the subchondral bone plate. Flow simulations showed a permeability decrease by about 2000-fold and solute transport simulations using a tracer (iodixanol, 1.5 kDa with a free diffusivity of 2.5 × 10-10 m2/s) showed diffusivity decrease by a factor of 1.5. Taken together, architecture of the nano-pores and the complex mineralization pattern in the osteochondral interface considerably impacts the cross-talk between cartilage and bone.


Doxorubicin resistance in breast cancer: A novel role for the human protein AHNAK.
Understanding the response of cancer cells to anti-cancer therapies is crucial to unraveling and preventing the development of therapeutic resistance. The human AHNAK protein is a giant scaffold protein implicated in several diverse cellular functions. The role of AHNAK in cancer is however unclear as the protein has previously been described as a tumor suppressor, as well as being essential for tumor metastasis and invasion, while also being implicated in selected chemotherapeutic responses. To clarify the role of AHNAK in cancer, we investigated the effect of doxorubicin treatment on AHNAK in doxorubicin-sensitive MCF-7 and doxorubicin-resistant MDA-MB-231 breast cancer cell lines, as well as in a tumor-bearing mouse model. The role of AHNAK in the cellular response of breast cancer cells to doxorubicin was also investigated. We report here, for the first time, an association between AHNAK and resistance to doxorubicin. While treatment with doxorubicin modulated AHNAK protein expression both in vitro and in vivo in a dose-dependent manner, no changes in its cellular localization were observed. AHNAK knockdown prevented doxorubicin-induced modulation of cleaved caspase 7 protein expression and cell cycle arrest, while its overexpression decreased cleaved caspase 7 and cleaved PARP levels and induced S-phase arrest, changes that were comparable to the effects of doxorubicin. This novel association was restricted to doxorubicin-resistant cells, implicating the protein in therapeutic resistance. These findings confirm that AHNAK does indeed function in the chemotherapeutic response of breast cancer cells while also emphasizing the need for further investigation into potential implications for AHNAK in terms of predicting and modulating treatment response.


Oxidative ecology of paternal care in wild smallmouth bass, Micropterus dolomieu.
Physiologically, oxidative stress is considered a homeostatic imbalance between reactive oxygen species production and absorption. From an ecological perspective, oxidative stress may serve as an important constraint to life-history traits, such as lifespan, reproduction and the immune system, and is gaining interest as a potential mechanism underlying life-history trade-offs. Of late, there has been much interest in understanding the role of oxidative stress in the ecology of wild animals, particularly during challenging periods such as reproduction. Here, we used a long-term study population of a fish with sole-male parental care, the smallmouth bass, Micropterus dolomieu, to examine the associations among oxidative stress indicators and life-history variables in nest-guarding males. In addition, we investigated the potential role of oxidative stress as a physiological mediator of the life-history trade-off decision of paternal smallmouth bass to stay with or abandon their brood. We found that oxidative stress was significantly related to the life history of paternal smallmouth bass, such that older, larger fish with greater reproductive experience and larger broods nesting in cooler water temperatures had lower levels of oxidative stress. However, we found no significant correlation between oxidative stress and nesting success, suggesting that oxidative stress may not be involved in the decision of male smallmouth bass to abandon their brood. Wild fish have been relatively understudied in the emerging field of oxidative ecology, and this study makes noteworthy contributions by revealing interesting connections between the life histories of paternal smallmouth bass and their oxidative status.


The Hidden Side of Zero Tolerance Policies: The African American Perspective.
Several papers have documented the disproportionate representation of African Americans in school discipline and incarceration settings as a result of zero tolerance policies. In 2009, a federal study of the Chicago Public School system found African American boys represented 23 percent of the school age population, 44 percent of students who were suspended, and 61 percent of students who were expelled within the 2007 school year. Twenty years after the implementation of the Anti-Drug Abuse Acts of 1986 and 1988, studies show African Americans comprised a startling 74 percent of those incarcerated for drug offenses despite being only 15 percent of America's drug users. Despite overwhelming evidence that suggests African Americans are adversely affected by zero tolerance policies, African American perceptions of zero tolerance policies remain relatively hidden in the literature. The current review seeks to explore a seemingly bidirectional process that involves how zero tolerance impacts African Americans and how African Americans perceive zero tolerance policies.


Cannabinoid signalling in TNF-alpha induced IL-8 release.
The molecular events mediating the immunomodulatory properties of cannabinoids have remained largely unresolved. We have therefore investigated the molecular mechanism(s) through which R-(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl] pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-napthanlenyl) methanone (WIN55212-2) modulate production of interleukin-8 (IL-8) in HT-29 cells. Release of IL-8 induced by tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme-linked immunosorbent assay (ELISA). Changes in expression of inhibitory kappa B (IkappaB) were monitored by Western blotting and activation of nuclear factor-kappa B (NF-kappaB) was determined in electrophoretic mobility shift assay (EMSAs). TNF-alpha induced release of IL-8 was inhibited by WIN55212-2 which also blocked the degradation of IkappaB-alpha and activation of NF-kappaB induced by TNF-alpha. These data provide strong evidence that WIN55212-2 may modulate IL-8 release by negatively regulating the signaling cascade leading to the activation of NF-kappaB. These findings highlight a potential mechanism for the immunomodulatory properties of cannabinoids and contribute towards acquiring a clear understanding of the role of cannabinoids in inflammation.


Variation in nephrologist visits to patients on hemodialysis across dialysis facilities and geographic locations.
Geographic and other variations in medical practices lead to differences in medical costs, often without a clear link to health outcomes. This work examined variation in the frequency of physician visits to patients receiving hemodialysis to measure the relative importance of provider practice patterns (including those patterns linked to geographic region) and patient health in determining visit frequency. This work analyzed a nationally representative 2006 database of patients receiving hemodialysis in the United States. A variation decomposition analysis of the relative importance of facility, geographic region, and patient characteristics--including demographics, socioeconomic status, and indicators of health status--in explaining physician visit frequency variation was conducted. Finally, the associations between facility, geographic and patient characteristics, and provider visit frequency were measured using multivariable regression. Patient characteristics accounted for only 0.9% of the total visit frequency variation. Accounting for case-mix differences, patients' hemodialysis facilities explained about 24.9% of visit frequency variation, of which 9.3% was explained by geographic region. Visit frequency was more closely associated with many facility and geographic characteristics than indicators of health status. More recent dialysis initiation and recent hospitalization were associated with decreased visit frequency. In hemodialysis, provider visit frequency depends more on geography and facility location and characteristics than patients' health status or acuity of illness. The magnitude of variation unrelated to patient health suggests that provider visit frequency practices do not reflect optimal management of patients on dialysis.


Nanopore film based enrichment and quantification of low abundance hepcidin from human bodily fluids.
Endogenous peptides that represent biological and pathological information of disease have attracted interest for diagnosis. However, the extraction of those low abundance peptides is still a challenge because of the complexity of human bodily fluids (HBF). Hepcidin, a peptide hormone, has been recognized as a biomarker for iron-related diseases. There is no rapid and reliable way to enrich them from HBF. Here we describe a peptide extraction approach based on nanoporous silica thin films to successfully detect hepcidin from HBF. Cooperative functions of nanopore to biomolecule, including capillary adsorption, size-exclusion and electrostatic interaction, were systematically investigated to immobilize the target peptide. To promote this new approach to clinical practices, we further applied it to successfully assay the hepcidin levels in HBF provided by healthy volunteers and patients suffering from inflammation. Our finding provides a high-throughput, rapid, label-free and cost-effective detection method for capturing and quantifying low abundance peptides from HBF. Diagnosing diseases with low concentration peptide biomarkers remains challenging. This team of authors describes a peptide extraction approach based on nanoporous silica thin films to successfully detect low concentrations of hepcidin from human body fluids collected from 119 healthy volunteers and 19 inflammation patients.


Organization and regulation of mitochondrial respiration in plants.
Mitochondrial respiration in plants provides energy for biosynthesis, and its balance with photosynthesis determines the rate of plant biomass accumulation. We describe recent advances in our understanding of the mitochondrial respiratory machinery of cells, including the presence of a classical oxidative phosphorylation system linked to the cytosol by transporters, discussed alongside nonphosphorylating (and, therefore, non-energy conserving) bypasses that alter the efficiency of ATP synthesis and play a role in oxidative stress responses in plants. We consider respiratory regulation in the context of the contrasting roles mitochondria play in different tissues, from photosynthetic leaves to nutrient-acquiring roots. We focus on the molecular nature of this regulation at transcriptional and post-transcriptional levels that allow the respiratory apparatus of plants to help shape organ development and the response of plants to environmental stress. We highlight the challenges for future research considering spatial and temporal changes of respiration in response to changing climatic conditions.


A rapid and computationally inexpensive method to virtually implant current and next-generation stents into subject-specific computational fluid dynamics models.
Computational modeling is often used to quantify hemodynamic alterations induced by stenting, but frequently uses simplified device or vascular representations. Based on a series of Boolean operations, we developed an efficient and robust method for assessing the influence of current and next-generation stents on local hemodynamics and vascular biomechanics quantified by computational fluid dynamics. Stent designs were parameterized to allow easy control over design features including the number, width and circumferential or longitudinal spacing of struts, as well as the implantation diameter and overall length. The approach allowed stents to be automatically regenerated for rapid analysis of the contribution of design features to resulting hemodynamic alterations. The applicability of the method was demonstrated with patient-specific models of a stented coronary artery bifurcation and basilar trunk aneurysm constructed from medical imaging data. In the coronary bifurcation, we analyzed the hemodynamic difference between closed-cell and open-cell stent geometries. We investigated the impact of decreased strut size in stents with a constant porosity for increasing flow stasis within the stented basilar aneurysm model. These examples demonstrate the current method can be used to investigate differences in stent performance in complex vascular beds for a variety of stenting procedures and clinical scenarios.


A hybrid chip based on aerodynamics and electrostatics for the size-dependent classification of ultrafine and nano particles.
Conventional virtual impactors experience a large pressure drop when they classify particles according to size, in particular ultrafine particles smaller than 100 nm in diameter. Therefore, most virtual impactors have been used to classify particles larger than 100 nm. Their cut-off diameters are also fixed by the geometry of their flow channels. In the proposed virtual impactor, particles smaller than 100 nm are accelerated by applying DC potentials to an integrated electrode pair. By the electrical acceleration, the large pressure drop could be significantly decreased and new cut-off diameters smaller than 100 nm could be successfully added. The geometric cut-off diameter (GCD) of the proposed virtual impactor was designed to be 1.0 microm. Performances including the GCD and wall loss were examined by classifying dioctyl sebacate of 100 to 600 nm in size and carbon particles of 0.6 to 10 microm in size. The GCD was measured to be 0.95 microm, and the wall loss was highest at 1.1 microm. To add new cut-off diameters, monodisperse NaCl particles ranging from 15 to 70 nm were classified using the proposed virtual impactor with applying a DC potential of 0.25 to 3.0 kV. In this range of the potential, the new cut-off diameters ranging from 15 to 35 nm was added.


CD3+, CD56+ aggressive variant of large granular lymphocyte leukemia.
Clonal expansions of CD3+ large granular lymphocytes (LGL) have been classified as T-LGL leukemia. The majority of patients with T-LGL leukemia have a chronic disease (years) manifested often by severe neutropenia, rheumatoid arthritis, and mild-to-moderate splenomegaly. The characteristic phenotype of the leukemic LGL is CD3+, CD8+, CD16+, CD57+, and CD56-. In this report we describe an aggressive variant of T-LGL leukemia in which leukemic LGL also expressed CD56, as identified by two-color flow-cytometry analysis. In contrast to the chronic nature typical of T-LGL leukemia, these patients presented with a severe systemic illness that was rapidly progressive and resistant to treatment. Atypical clinical features included rapidly increasing spleen size to massive proportions, extensive lymphadenopathy, and the presence of B symptoms (fever, nightsweats, weight loss). Hematologic and pathologic features were also unusual for T-LGL leukemia. These patients had very high LGL counts at diagnosis (range 11,692 to 26,312 microL), which increased rapidly despite treatment. Histopathologic examination of splenic sections showed extensive infiltration of red pulp cords and sinuses by leukemic cells with atrophy of the white pulp. These clinicopathologic features are similar to those described for patients with natural killer cell (NK)-LGL leukemia, whose cells are also CD56+. However, unlike NK-LGL leukemia, we could not show a direct pathogenic role for Epstein-Barr virus (EBV), as Southern-blot analyses using an EBV-joined termini probe were negative in these patients. Our findings suggest that CD3+, CD56+ LGL leukemia is a distinct clinicopathologic entity separate from the usual CD3+, CD56- T-LGL leukemia. The expression on leukemic LGL of CD56, an adhesion molecule, may determine the aggressive biologic nature of this newly described disease.


[Production chain supply management for public hospitals: a logistical approach to healthcare].
Despite their importance for hospital operations, discussions of healthcare organization logistics and supply and materials management are notably lacking in Brazilian literature. This paper describes a methodology for organizing the supply of medical materials in public hospitals, based on an action-research approach. Interventions were based on the assumption that a significant portion of problems in Brazil's National Health System (SUS) facilities derive from the fact that their clinical and administrative departments do not see themselves as belonging to the same production chain - neither the hospital nor the supply department is aware of what the other produces. The development of the methodology and its main steps are presented and discussed, against a background of recent literature and total quality and supply chain management concepts.


Health impact and damage cost assessment of pesticides in Europe.
Health impacts from pesticide use are of continuous concern in the European population, requiring a constant evaluation of European pesticide policy. However, health impacts have never been quantified accounting for specific crops contributing differently to overall human exposure as well as accounting for individual substances showing distinct environmental behavior and toxicity. We quantify health impacts and related damage costs from exposure to 133 pesticides applied in 24 European countries in 2003 adding up to almost 50% of the total pesticide mass applied in that year. Only 13 substances applied to 3 crop classes (grapes/vines, fruit trees, vegetables) contribute to 90% of the overall health impacts of about 2000 disability-adjusted life years in Europe per year corresponding to annual damage costs of 78 million Euro. Considering uncertainties along the full impact pathway mainly attributable to non-cancer dose-response relationships and residues in treated crops, we obtain an average burden of lifetime lost per person of 2.6 hours (95% confidence interval between 22 seconds and 45.3 days) or costs per person over lifetime of 12 Euro (95% confidence interval between 0.03 Euro and 5142 Euro), respectively. 33 of the 133 assessed substances accounting for 20% of health impacts in 2003 are now banned from the European market according to current legislation. The main limitation in assessing human health impacts from pesticides is related to the lack of systematic application data for all used substances. Since health impacts can be substantially influenced by the choice of pesticides, the need for more information about substance application becomes evident.


Transcorneal oxygenation of the preretinal vitreous.
Transcorneal oxygenation is a promising approach to the treatment of various anterior ocular diseases, but its effect on the vitreous and retina is uncertain. The purpose of this study was to evaluate the ocular conditions necessary to permit transcorneal oxygenation of the preretinal vitreous in the rabbit eye. Oxygen at atmospheric pressure was supplied via a goggle to the eye while preretinal oxygen tensions were measured with a minimally invasive technique: fluorine-19 (19F) nuclear magnetic resonance spectroscopy of a small preretinal droplet of perfluorotributylamine. Experiments were performed on lensectomized eyes with intact posterior lens capsules (group 1) or in eyes that had also undergone posterior capsular discission (group 2) and mechanical vitrectomy (group 3). To better understand the results of the oxygenation studies, a water-soluble, freely diffusible contrast agent was applied topically to the cornea, and its penetration into the aqueous and vitreous was studied using T1-weighted magnetic resonance imaging. Preretinal PO2 was significantly increased after 30 minutes of transcorneal oxygenation in group 3 eyes (P = .002). In contrast, no change was observed in the other groups despite 30 minutes (group 1) or 3 hours (group 2) of oxygen exposure. The imaging studies demonstrated that contrast penetration of the vitreous was uniform and rapid in eyes in group 3, consistent with a fluid flow-dominated process. A much slower process of anterior and midvitreous enhancement was observed in groups 1 and 2 that was consistent with contrast diffusion. Because of bulk fluid movement, transcorneal oxygenation of the retina is feasible in single-compartment (vitrectomized, lensectomized) eyes within a reasonable period (30 minutes). This finding could prove useful in the treatment of retinal diseases in which hypoxia may play a role.


Large-scale genomic studies reveal central role of ABO in sP-selectin and sICAM-1 levels.
P-selectin and intercellular adhesion molecule-1 (ICAM-1) participate in inflammatory processes by promoting adhesion of leukocytes to vascular wall endothelium. Their soluble levels have been associated with adverse cardiovascular events. To identify loci affecting soluble levels of P-selectin (sP-selectin) and ICAM-1 (sICAM-1), we performed a genome-wide association study in a sample of 4115 (sP-selectin) and 9813 (sICAM-1) individuals of European ancestry as a part of The Cohorts for Heart and Aging Research in Genome Epidemiology consortium. The most significant SNP association for sP-selectin was within the SELP gene (rs6136, P = 4.05 x 10(-61)) and for sICAM-1 levels within the ICAM-1 gene (rs3093030, P = 3.53 x 10(-23)). Both sP-selectin and sICAM-1 were associated with ABO gene variants (rs579459, P = 1.86 x 10(-41) and rs649129, P = 1.22 x 10(-15), respectively) and in both cases the observed associations could be accounted for by the A1 allele of the ABO blood group. The absence of an association between ABO blood group and platelet-bound P-selectin levels in an independent subsample (N = 1088) from the ARIC study, suggests that the ABO blood group may influence cleavage of the P-selectin protein from the cell surface or clearance from the circulation, rather than its production and cellular presentation. These results provide new insights into adhesion molecule biology.


A new multidimensional population health indicator for policy makers: absolute level, inequality and spatial clustering - an empirical application using global sub-national infant mortality data.
The need for a multidimensional measure of population health that accounts for its distribution remains a central problem to guide the allocation of limited resources. Absolute proxy measures, like the infant mortality rate (IMR), are limited because they ignore inequality and spatial clustering. We propose a novel, three-part, multidimensional mortality indicator that can be used as the first step to differentiate interventions in a region or country. The three-part indicator (MortalityABC index) combines absolute mortality rate, the Theil Index to calculate mortality inequality and the Getis-Ord G statistic to determine the degree of spatial clustering. The analysis utilises global sub-national IMR data to empirically illustrate the proposed indicator. The three-part indicator is mapped globally to display regional/country variation and further highlight its potential application. Developing countries (e.g. in sub-Saharan Africa) display high levels of absolute mortality as well as variable mortality inequality with evidence of spatial clustering within certain sub-national units ("hotspots"). Although greater inequality is observed outside developed regions, high mortality inequality and spatial clustering are common in both developed and developing countries. Significant positive correlation was observed between the degree of spatial clustering and absolute mortality. The proposed multidimensional indicator should prove useful for spatial allocation of healthcare resources within a country, because it can prompt a wide range of policy options and prioritise high-risk areas. The new indicator demonstrates the inadequacy of IMR as a single measure of population health, and it can also be adapted to lower administrative levels within a country and other population health measures.


Material parameter estimation with terahertz time-domain spectroscopy.
Imaging systems based on terahertz (THz) time-domain spectroscopy offer a range of unique modalities owing to the broad bandwidth, subpicosecond duration, and phase-sensitive detection of the THz pulses. Furthermore, the possibility exists for combining spectroscopic characterization or identification with imaging because the radiation is broadband in nature. To achieve this, we require novel methods for real-time analysis of THz waveforms. This paper describes a robust algorithm for extracting material parameters from measured THz waveforms. Our algorithm simultaneously obtains both the thickness and the complex refractive index of an unknown sample under certain conditions. In contrast, most spectroscopic transmission measurements require knowledge of the sample's thickness for an accurate determination of its optical parameters. Our approach relies on a model-based estimation, a gradient descent search, and the total variation measure. We explore the limits of this technique and compare the results with literature data for optical parameters of several different materials.


High Prevalence of Diabetes Mellitus in a Cohort of Patients with Chronic Obstructive Pulmonary Disease in Trinidad, West Indies.
Both chronic obstructive pulmonary disease (COPD) and diabetes mellitus (DM) are highly prevalent in Trinidad, West Indies. Our objective was to evaluate the prevalence of DM in a cohort of Trinidadian patients with COPD and investigate the possible impact of diabetes on COPD using standard outcome measures, that is, lung function, exacerbations, quality of life and depression questionnaires, as well as mortality. This was a cross-sectional follow-up study utilizing a cohort of 105 patients from chest clinics in the three major general hospitals in Trinidad. Diabetes was diagnosed based on a glycated hemoglobin (HbA1c) level of ≥6.5% (or a prior self-reported history), and for pre-diabetes, of 5.7%-6.4%. Of 105 patients, 40% fulfilled the criteria for diabetes and 40% for pre-diabetes. Of those diagnosed with diabetes, 38% obtained this diagnosis de novo. A history of intravenous corticosteroid use was associated with higher HbA1c levels (p=0.043) upon diagnosis. The percentage of predicted forced vital capacity was negatively related to HbA1c (p=0.033), but those with diabetes also had a greater body mass index (p=0.001). After a 1-year follow-up, mortality was significantly greater among patients with diabetes (p=0.026). Patients with at least one exacerbation in the past year or poorer lung function parameters had worse quality of life (p≤0.040) and depression (p≤0.018) scores. Notably, 31.4% of the total cohort exhibited clinically significant depression scores. This study revealed that a high proportion of COPD patients in tertiary care had diabetes or pre-diabetes.


Analysis of PHOTOPERIOD SENSITIVITY5 sheds light on the role of phytochromes in photoperiodic flowering in rice.
A great number of plants synchronize flowering with day length. In rice (Oryza sativa), photoperiod is the primary environmental cue that triggers flowering. Here, we show that the s73 mutant, identified in a gamma-irradiated Bahia collection, displays early flowering and photoperiodic insensitivity due to a null mutation in the PHOTOPERIOD SENSITIVITY5 (SE5) gene, which encodes an enzyme implicated in phytochrome chromophore biosynthesis. s73 mutant plants show a number of alterations in the characteristic diurnal expression patterns of master genes involved in photoperiodic control of flowering, resulting in up-regulation of the floral integrator Heading date3a (Hd3a). Early heading date1 (Ehd1), an additional rice floral activator, was also highly expressed in the s73 mutant, suggesting that SE5 represses Ehd1 in wild-type plants. Silencing of Ehd1 in both Bahia and s73 backgrounds indicated that SE5 regulates Ehd1 expression. The data also indicate that SE5 confers photoperiodic sensitivity through regulation of Hd1. These results provide direct evidence that phytochromes inhibit flowering by affecting both Hd1 and Ehd1 flowering pathways.


Isolation and characterization of chemotaxis mutants and genes of Pseudomonas aeruginosa.
Two chemotaxis-defective mutants of Pseudomonas aeruginosa, designated PC1 and PC2, were selected by the swarm plate method after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. These mutants were fully motile but incapable of swarming, suggesting that they had a defect in the intracellular signalling pathway. Computer-assisted capillary assays confirmed that they failed to show behavioral responses to chemical stimuli, including peptone, methyl thiocyanate, and phosphate. Two chemotaxis genes were cloned by phenotypic complementation of PC1 and PC2. From nucleotide sequence analysis, one gene was found to encode a putative polypeptide that was homologous to the enteric CheZ protein, while the other gene was cheY, which had been previously reported (M. N. Starnbach and S. Lory, Mol. Microbiol. 6:459-469, 1992). Deletion and complementation analysis showed that PC1 was a cheY mutant, whereas PC2 had a double mutation in the cheY and cheZ genes. A chromosomal cheZ mutant, constructed by inserting a kanamycin resistance gene cassette into the wild-type gene, changed its swimming direction much more frequently than did wild-type strain PAO1. In contrast, cheY mutants were found to rarely reverse their swimming directions.


Stent Retriever-Assisted Mechanical Thrombectomy for Acute Basilar Artery Occlusion: Single US Institution Experience.
Acute basilar artery occlusion causes devastating strokes that carry high mortality and morbidity. To report the outcomes of mechanical thrombectomy in the posterior circulation with a focus on safety and efficacy of stent retrievers. We retrospectively reviewed our endovascular database for all patients treated with stent retrievers for posterior circulation stroke between June 2012 and June 2014. Twelve patients were identified. The following data were analyzed: thrombus location, previous stroke or transient ischemic attack, thrombus etiology, comorbidities, time from presentation to initiation of endovascular treatment, time from start of angiography to revascularization, and whether intravenous tissue plasminogen activator was administered pre-thrombectomy. Outcome was considered poor when modified Rankin Scale score was >2. Mean patient age was 63.42 years (median, 64.5; range, 28-83 years); 7 were women. Successful recanalization (Thrombolysis in Cerebral Infarction grade 2b or 3) was achieved in 11 of 12 patients (91.7%). Mean discharge modified Rankin Scale score was 2.3 (median, 2.0; standard deviation 1.96; range, 0-6), with a favorable discharge outcome in 9 of 12 (75%) patients. Two patients died as inpatients. Mean follow-up modified Rankin Scale score was 1.4 (median, 1.00; standard deviation 1.075; range, 0-4). Good outcome was achieved in 9 of 10 (90%) patients at last follow-up (mean follow-up duration, 132.42 days [median, 90.50; standard deviation 80.2; range, 8-378 days]). Our single-institution study has shown that good clinical outcomes and successful recanalization with acceptable mortality can be achieved with current stent retrievers.


AUTOMATIC AIRWAY ANALYSIS FOR GENOME-WIDE ASSOCIATION STUDIES IN COPD.
We present an image pipeline for airway phenotype extraction suitable for large-scale genetic and epidemiological studies including genome-wide association studies (GWAS) in Chronic Obstructive Pulmonary Disease (COPD). We use scale-space particles to densely sample intraparenchymal airway locations in a large cohort of high-resolution CT scans. The particle methodology is based on a constrained energy minimization problem that results in a set of candidate airway points situated in both physical space and scale. Those points are further clustered using connected components filtering to increase their specificity. Finally, we use the particle locations to perform airway wall detection using an edge detector based on the zero-crossing of the second order derivative. Given the airway wall locations, we compute three phenotypes for airway disease: wall thickening (Pi10,WA%) and luminal remodeling (P%). We validate the airway extraction technique and present results in 2,500 scans for the association of the extracted phenotypes with clinical outcomes that will be deployed as part of the COPDGene study GWAS analysis.


Clinical heterogeneity of familial colorectal cancer and its influence on screening protocols.
The age at onset of non-polyposis colorectal cancer (CRC) was investigated in 49 families with at least three relatives affected in two successive generations. Forty one of these families satisfy the accepted minimum criteria for hereditary non-polyposis CRC. The remaining eight families were distinguished by a late age of disease onset and, hence, could not be included in the group. The condition in these latter families has been designated provisionally, as late onset familial CRC. The hereditary non-polyposis CRC families include 194 patients, 110 men and 84 women (mean age at diagnosis: 44 years; range: 16-74 years). Ninety two per cent of the patients were diagnosed by age 60. Colorectal cancer was diagnosed at progressively earlier ages in successive generations (p < 0.0005). The late onset CRC families comprised 30 patients, 20 men and 10 women (mean age at diagnosis: 62 years; range: 51-77 years). Fifty eight per cent of the CRCs in the patients belonging to the hereditary non-polyposis CRC families were located in the right colon compared with 13% in the late onset familial CRC group (p < 0.001). Multiple CRCs were found in 23% of the cases in the former but in only 3% in the latter families (p < 0.05). Adenomas associated with CRC were reported in 14.5% of the cases in the hereditary non-polyposis CRC families and in 30% of the cases in the late onset familial CRC group (p = NS). Extracolonic cancers, frequently encountered in hereditary non-polyposis CRC, were not found in the late onset CRC families. These findings indicate that there is a distinct clinical entity of non-polyposis CRC in which colorectal cancer develops at a more advanced age than in hereditary non-polyposis CRC. Future molecular genetic studies should confirm this hypothesis. In the meantime, recognition of these late onset familial CRC families, which will rest on clinical observations, is important because of the implications for the screening protocol.


Molecular and biological factors in the prognosis of non-small cell lung cancer.
For patients with non-small cell lung cancer the tumour/node/metastasis (TNM) staging system and other conventional prognostic factors fail to predict the outcome of treatment and survival accurately. New prognostic factors are urgently needed to improve understanding of the biological behaviour of the different subtypes of non-small cell lung cancer and to recognize patients with a good or poor prognosis. This review will focus on molecular and biological factors published in the English language literature between 1988 and 1994. To be included in this survey, the predictive value of a specific prognostic factor had to be confirmed by multivariate analysis in at least two different studies. Blood group antigen expression, ras oncogenes, microvessel density, and factors reflecting the proliferative state of the tumour may be important determinants of outcome of treatment. The search for new determinants of prognosis has provided insight in the complex tumour biology of non-small cell lung cancer and indicated possible targets for tumour therapy. Several promising prognostic factors have now been recognized. To validate these factors, prospective studies of a large patient population are needed. This ultimately serves the recognition of subsets of patients who may benefit from adjuvant therapy.


Transconjunctival Müller muscle recession with levator disinsertion for correction of eyelid retraction associated with thyroid-related orbitopathy.
To evaluate the efficacy of transconjunctival Müller muscle recession and graded levator disinsertion for eyelid retraction in patients with thyroid-related orbitopathy (TRO). Retrospective consecutive case series. Medical record review of 78 TRO patients (107 eyelids) who underwent surgery for upper eyelid retraction in a 5-year period was performed. Main outcome measures were anatomic and functional success, minimal reflex distance (MRD), lagophthalmos, eyelid asymmetry, and patient discomfort. One hundred seven eyelid retraction surgeries were performed on 78 TRO patients (63 women, mean age 49 years); mean follow-up time was 16.7 months. Upper eyelid position, lagophthalmos, exposure keratopathy, and patients' discomfort markedly improved after surgery (P < .001). Marginal reflex distance (MRD1) decreased an average of 2.6 mm from 6 mm pre-operatively to 3.4 mm post-operatively (P < .001); lagophthalmos decreased an average of 0.6 mm from 1.3 mm pre-operatively to 0.4 mm post-operatively (P = .006) Failure rate was 8.4%, most improved with a second surgery. Overcorrection was noticed in three cases (2.8%). Eyelid asymmetry improved from a mean of 1.0 mm pre-operatively to 0.4 mm post-operatively (P = .001); more than 80% of patients showed eyelid asymmetry of 1 mm or less. Transconjunctival Müller muscle and levator recession is safe and effective in correction of mild, moderate, or severe eyelid retraction in TRO patients. The failure rate is less than 10% and may be addressed by a second surgery.


Thresholding of statistical maps in functional neuroimaging using the false discovery rate.
Finding objective and effective thresholds for voxelwise statistics derived from neuroimaging data has been a long-standing problem. With at least one test performed for every voxel in an image, some correction of the thresholds is needed to control the error rates, but standard procedures for multiple hypothesis testing (e.g., Bonferroni) tend to not be sensitive enough to be useful in this context. This paper introduces to the neuroscience literature statistical procedures for controlling the false discovery rate (FDR). Recent theoretical work in statistics suggests that FDR-controlling procedures will be effective for the analysis of neuroimaging data. These procedures operate simultaneously on all voxelwise test statistics to determine which tests should be considered statistically significant. The innovation of the procedures is that they control the expected proportion of the rejected hypotheses that are falsely rejected. We demonstrate this approach using both simulations and functional magnetic resonance imaging data from two simple experiments.


Finite element modeling to compare craniocervical motion in two age-matched pediatric patients without or with Down syndrome: implications for the role of bony geometry in craniocervical junction instability.
Instability of the craniocervical junction (CCJ) is a well-known finding in patients with Down syndrome (DS); however, the relative contributions of bony morphology versus ligamentous laxity responsible for abnormal CCJ motion are unknown. Using finite element modeling, the authors of this study attempted to quantify those relative differences. Two CCJ finite element models were created for age-matched pediatric patients, a patient with DS and a control without DS. Soft tissues and ligamentous structures were added based on bony landmarks from the CT scans. Ligament stiffness values were assigned using published adult ligament stiffness properties. Range of motion (ROM) testing determined that model behavior most closely matched pediatric cadaveric data when ligament stiffness values were scaled down to 25% of those found in adults. These values, along with those assigned to the other soft-tissue materials, were identical for each model to ensure that the only variable between the two was the bone morphology. The finite element models were then subjected to three types of simulations to assess ROM, anterior-posterior (AP) translation displacement, and axial tension. The DS model exhibited more laxity than the normal model at all levels for all of the cardinal ROMs and AP translation. For the CCJ, the flexion-extension, lateral bending, axial rotation, and AP translation values predicted by the DS model were 40.7%, 52.1%, 26.1%, and 39.8% higher, respectively, than those for the normal model. When simulating axial tension, the soft-tissue structural stiffness values predicted by the DS and normal models were nearly identical. The increased laxity exhibited by the DS model in the cardinal ROMs and AP translation, along with the nearly identical soft-tissue structural stiffness values exhibited in axial tension, calls into question the previously held notion that ligamentous laxity is the sole explanation for craniocervical instability in DS.


Denoising analysis of Hadamard transform spectrometry.
We discuss denoising in Hadamard transform spectrometry (HTS) in terms of sensor noise, photon noise, and the sparsity of the source. An analysis based on spectra classification is proposed to estimate the signal-to-noise ratio (SNR) of both HTS and slit-based spectrometry. In contrast with previous theory, it is shown that HTS can improve the sensitivity of the sensor and that HTS outperforms slit-based spectrometry when the signal is dominated by photon noise and the source is sparse. Numerical simulations show that HTS is a good method for improving the poor SNR associated with weak or sparse signals.


Multiangle Imaging Spectroradiometer: optical characterization of the calibration panels.
The reflectance properties of an engineering model (EM) of the Spectralon panel intended for use within an on-board calibrator (OBC) on the NASA Multiangle Imaging Spectroradiometer (MISR) instrument have been fully characterized with regard to panel uniformity and isotropy in response to three incident laser wavelengths of 442, 632.8, and 859.9 nm. A regional variation in the relative bidirectional reflectance factor (RBRF) across the surface of the EM panel, which contributes to spatial nonuniformity at the +/-2% level, has been measured at all three laser wavelengths. Further, a reflectance anisotropy has been identified. The mechanism causing these departures from the ideal Lambertian surface may originate in the sanding of the Spectralon surface in the final stage of preparation. This supposition is corroborated by measurements made on a pressed polytetrafluoroethylene (PTFE) panel in which a greatly reduced anisotropy in panel RBRF is measured. The EM panel RBRF exhibits a deviation from Lambertian characteristics as an off-specular peak in the forward scattering direction. A common crossover point at an angle of reflection of approximately 37 degrees at which the BRF is constant within +/-0.4% for an illumination angle range of theta(i) = 30 degrees-60 degrees is observed at all three wavelengths. Two Spectralon protoflight panels that were fabricated after the EM was studied were also the subject of a uniformity study over part of the area of the Spectralon panels at the 442-nm wavelength. The analysis indicated that the panel uniformity satisfies the +/-0.5% criterion, which indicates improved panel preparation. However, the off-specular peak in the forward scattering direction was essentially unchanged, with the crossover point at approximately 37 degrees.


Atopy among schoolchildren in northern and southern Sweden in relation to pet ownership and early life events.
Studies have suggested a higher prevalence of asthma and allergies in northern, as compared to southern, Scandinavia. The aim of this study was to evaluate regional differences in atopy in relation to pet ownership and certain early life events among schoolchildren (n=2108) aged 10-11 years from Linköping in southern Sweden and Ostersund in northern Sweden. The parents completed a questionnaire, comprising questions on home environment, heredity, socio-economic conditions, and the core questions on symptoms from the International Study of Asthma and Allergies in Childhood. The children were skin-prick tested to eight common inhalant allergens. Information on maternal smoking habits, gestational age, and anthropometric measures were obtained from the Swedish Medical Birth Registry. The prevalence of atopic symptoms and sensitization to pollen were similar in Ostersund and in Linköping. A higher prevalence of sensitization to animal dander among children in Ostersund could be linked to a higher occurrence of pets in the community. Current cat ownership was related to less sensitivity to cat allergen but only in children with an atopic heredity. Ponderal index >30 kg/m3 was related to an increased risk of atopic sensitization, both in Linköping (adjusted odds ratio 2.1; 95% confidence interval 1.1-4.0) and in Ostersund (adjusted odds ratio 2.0; 95% confidence interval 1.1-3.5). Maternal smoking during pregnancy was related to an increased risk of atopic sensitization among children in Linköping, whereas current smoking was associated with a decreased risk of sensitization in Ostersund. In conclusion, we demonstrated that a high occurrence of pets in the community was associated with sensitization, whereas atopic symptoms were essentially unaffected. This study has also suggested an association between body size at birth and atopic sensitization at 10-11 years of age.


The Etiology of Genital Ulcer Disease and Coinfections With Chlamydia trachomatis and Neisseria gonorrhoeae in Zimbabwe: Results From the Zimbabwe STI Etiology Study.
In many countries, sexually transmitted infections (STIs) are treated syndromically. Thus, patients diagnosed as having genital ulcer disease (GUD) in Zimbabwe receive a combination of antimicrobials to treat syphilis, chancroid, lymphogranuloma venereum (LGV), and genital herpes. Periodic studies are necessary to assess the current etiology of GUD and assure the appropriateness of current treatment guidelines. We selected 6 geographically diverse clinics in Zimbabwe serving high numbers of STI cases to enroll men and women with STI syndromes, including GUD. Sexually transmitted infection history and risk behavioral data were collected by questionnaire and uploaded to a Web-based database. Ulcer specimens were obtained for testing using a validated multiplex polymerase chain reaction (M-PCR) assay for Treponema pallidum (TP; primary syphilis), Haemophilus ducreyi (chancroid), LGV-associated strains of Chlamydia trachomatis, and herpes simplex virus (HSV) types 1 and 2. Blood samples were collected for testing with HIV, treponemal, and nontreponemal serologic assays. Among 200 GUD patients, 77 (38.5%) were positive for HSV, 32 (16%) were positive for TP, and 2 (1%) were positive for LGV-associated strains of C trachomatis. No H ducreyi infections were detected. No organism was found in 98 (49.5%) of participants. The overall HIV positivity rate was 52.2% for all GUD patients, with higher rates among women compared with men (59.8% vs 45.2%, P < 0.05) and among patients with HSV (68.6% vs 41.8%, P < 0.0001). Among patients with GUD, 54 (27.3%) had gonorrhea and/or chlamydia infection. However, in this latter group, 66.7% of women and 70.0% of men did not have abnormal vaginal or urethral discharge on examination. Herpes simplex virus is the most common cause of GUD in our survey, followed by T. pallidum. No cases of chancroid were detected. The association of HIV infections with HSV suggests high risk for cotransmission; however, some HSV ulcerations may be due to HSV reactivation among immunocompromised patients. The overall prevalence of gonorrhea and chlamydia was high among patients with GUD and most of them did not meet the criteria for concomitant syndromic management covering these infections.


Targeting wild-type Erythrocyte receptors for Plasmodium falciparum and vivax Merozoites by Zinc Finger Nucleases In- silico: Towards a Genetic Vaccine against Malaria.
Malaria causes immense human morbidity and mortality globally. The plasmodium species vivax and falciparum cause over 75 % clinical malaria cases. Until now, gene-based strategies against malaria have only been applied to plasmodium species and their mosquito-vector. Merozoites of these two respective plasmodium species target and invade red blood cells (RBCs) by using the duffy antigen receptor for chemokines (DARC), and Sialic Acid (SLC4A1) residues of the O-linked glycans of Glycophorin A. RBCs of naturally selected duffy-negative blacks are resistant to P.vivax tropism. We hypothesized that artificial aberration of the host-pathway by target mutagenesis of either RBC -receptors, may abolish or reduce susceptibility of the host to malaria. As a first step towards the experimental actualization of these concepts, we aimed to identify zinc finger arrays (ZFAs) for constructing ZFNs that target genes of either wild-type host-RBC- receptors. In-Silico Gene & Genome Informatics Using the genomic contextual nucleotide-sequences of homo-sapiens darc and glycophorin-a, and the ZFN-consortia software- CoDA-ZiFiT-ZFA and CoDA-ZiFiT-ZFN: we identified 163 and over 1,000 single zinc finger arrays (sZFAs) that bind sequences within the genes for the two respective RBC-receptors. Second, 2 and 18 paired zinc finger arrays (pZFAs) that are precursors for zinc finger nucleases (ZFNs) capable of cleaving the genes for darc and glycophorin-a were respectively assembled. Third, a mega-BLAST evaluation of the genome-wide cleavage specificity of this set of ZFNs was done, revealing alternate homologous nucleotide targets in the human genome other than darc or glycophorin A. ZFNs engineered with these ZFA-precursors--with further optimization to enhance their specificity to only darc and glycophorin-a, could be used in constructing an experimental gene-based-malaria vaccine. Alternatively, meganucleases and transcription activator-like (TAL) nucleases that target conserved stretches of darc and glycophorin-a DNA may serve the purpose of abrogating invasion of RBCs by falciparam and vivax plasmodia species.


C-Myb(+) erythro-myeloid progenitor-derived fetal monocytes give rise to adult tissue-resident macrophages.
Although classified as hematopoietic cells, tissue-resident macrophages (MFs) arise from embryonic precursors that seed the tissues prior to birth to generate a self-renewing population, which is maintained independently of adult hematopoiesis. Here we reveal the identity of these embryonic precursors using an in utero MF-depletion strategy and fate-mapping of yolk sac (YS) and fetal liver (FL) hematopoiesis. We show that YS MFs are the main precursors of microglia, while most other MFs derive from fetal monocytes (MOs). Both YS MFs and fetal MOs arise from erythro-myeloid progenitors (EMPs) generated in the YS. In the YS, EMPs gave rise to MFs without monocytic intermediates, while EMP seeding the FL upon the establishment of blood circulation acquired c-Myb expression and gave rise to fetal MOs that then seeded embryonic tissues and differentiated into MFs. Thus, adult tissue-resident MFs established from hematopoietic stem cell-independent embryonic precursors arise from two distinct developmental programs.


Using the economics of certification to improve the safety and quality of male circumcision in developing countries: three models of implementation.
Although male circumcision (MC) has been a widespread practice in some regions, while relatively new in others, it has recently ascended in popularity as a HIV-reduction intervention, particularly in areas with high rates of HIV but low rates of MC. However, the uptake and potential effectiveness of MC may be hampered by uneven levels of provider training and procedure skill within developing country settings. Indeed, this procedure that is otherwise considered simple and safe has witnessed complication rates as high as 25-35% in some areas, leaving some men with irreversible injuries. To improve the transparency of procedure quality for prospective patients, I borrow from a classic economics approach and advocate a new application in the form of provider certification. Building on some experience in the healthcare systems and economic rationale of high-income counties, I explore the potential for certifying providers of MC in low-income countries and compare and contrast three models of implementation: government agency, private certifiers and private MC device manufacturers. The hope is that increased transparency of provider quality through any or all three types of certifying programmes can better assist local men as they navigate this otherwise complex and unclear medical care market. As more resources are being devoted to MC scale up, I argue that certification should be considered for incorporation as a means of complementing both current and future efforts in order to enhance the effectiveness of MC campaigns. The two models based on privatized certification, as opposed to having the local government underwrite the intervention, may prove most useful when public or philanthropic funding is volatile or incomplete for a given location. The timing for MC campaign adoption and desired speed of scale up may vary across countries in ways that international assistance efforts cannot always immediately and flexibly adapt to. As such, the role of the diverse MC provider marketplace and accompanying quality-revelation mechanisms may take on different levels of importance and expediency across settings as individual countries move forward with their respective HIV prevention campaigns. The subsequent challenge is to creatively design solutions that are sustainable and applicable within diverse host-country environments and expectations. This is where I believe some economic insights are currently lacking in the MC dialogue. Although I believe the three certification models exhibit much potential for enhancing medical care delivery in developing countries, they are not without their challenges, and implementation would not necessarily be a simple process. Local levels of medical knowledge, public and private resource constraints and the integrity of local business transactions and government practices would likely influence the nature and success of a certification intervention. However, with sufficient model adaptability and partnerships across public and private sectors, I argue that many of these implementation issues could be proactively addressed. Creative and careful certification structures should ultimately improve the MC circumstances across a variety of developing countries.


Tunable s-SNOM for Nanoscale Infrared Optical Measurement of Electronic Properties of Bilayer Graphene.
Here we directly probe the electronic properties of bilayer graphene using s-SNOM measurements with a broadly tunable laser source over the energy range from 0.3 to 0.54 eV. We tune an OPO/OPA system around the interband resonance of Bernal stacked bilayer graphene (BLG) and extract amplitude and phase of the scattered light. This enables us to retrieve and reconstruct the complex optical conductivity resonance in BLG around 0.39 eV with nanoscale resolution. Our technique opens the door toward nanoscopic noncontact measurements of the electronic properties in complex hybrid 2D and van der Waals material systems, where scanning tunneling spectroscopy cannot access the decisive layers.


Recruitment of CCR6-expressing Th17 cells by CCL20 secreted from plasmin-stimulated macrophages.
In the present study, monocyte-derived human macrophages were differentiated from buffy coats. Naïve CD4⁺ T-cells enriched from peripheral blood mononuclear cells using anti-CD4 magnetic beads and the autoMACS separation system were polarized under T-helper 17 (Th17)-promoting conditions for 6 days to get Th17 cells. The frequency of Th17 cell differentiation and the expression of C-C chemokine receptor type 6 (CCR6) on Th17 cells were investigated by flow cytometry. Plasmin-triggered induction of macrophage inflammatory protein-3alpha/C-C chemokine ligand 20 (CCL20) genes in macrophages was assessed by reverse transcription-polymerase chain reaction, and secreted protein levels were measured by enzyme-linked immunosorbent assay. Th17 cell migration induced by CCL20 secreted from plasmin-stimulated macrophages was tested in vitro by chemotaxis using a transwell system. These results demonstrate that plasmin triggers the expression of chemokine CCL20 messenger RNA and the release of CCL20 protein in human monocyte-derived macrophages, which critically depend on the proteolytic activity of plasmin and activation of p38 mitogen-activated protein kinase and nuclear factor-kappaB signaling pathways. Expression of CCR6 was detected on 87.23 ± 8.6% of Th17 cells in vitro. Similar to chemotaxis triggered by recombinant human CCL20, supernatants collected from plasmin-stimulated macrophage-induced chemotactic migration of Th17 cells, which could be inhibited by an anti-CCL20 neutralizing antibody. These results suggest that plasmin generated in inflamed tissues might elicit production of chemokine CCL20 by human macrophages leading to the recruitment of CCR6 positive Th17 cells to the inflammatory sites.


In situ hybridization of GAD mRNA in monkey and human brain: quantification at both regional and cellular levels.
GAD mRNA was detected in human and monkey brain postmortem by in situ hybridization with an [35S]-labelled copy RNA corresponding to a 2.7-kb fragment of the coding region of human GAD mRNA. A characteristic and reproducible pattern of hybridization was obtained with the anti-sense, but not the sense probe in both monkey and human brain. Microscopic examination of tissue sections showed that only neuronal perikarya, not glial cells, were labelled. The data confirm the heterogeneity of GAD mRNA distribution reported in rodent brain and non-human primate brain.


Effect of depression on diagnosis, treatment, and mortality of men with clinically localized prostate cancer.
Although demographic, clinicopathologic, and socioeconomic differences may affect treatment and outcomes of prostate cancer, the effect of mental health disorders remains unclear. We assessed the effect of previously diagnosed depression on outcomes of men with newly diagnosed prostate cancer. We performed a population-based observational cohort study using Surveillance, Epidemiology, and End Results-Medicare linked data of 41,275 men diagnosed with clinically localized prostate cancer from 2004 to 2007. We identified 1,894 men with a depressive disorder in the 2 years before the prostate cancer diagnosis and determined its effect on treatment and survival. Men with depressive disorder were older, white or Hispanic, unmarried, resided in nonmetropolitan areas and areas of lower median income, and had more comorbidities (P < .05 for all), but there was no variation in clinicopathologic characteristics. In adjusted analyses, men with depressive disorder were more likely to undergo expectant management for low-, intermediate-, and high-risk disease (P ≤ .05, respectively). Conversely, depressed men were less likely to undergo definitive therapy (surgery or radiation) across all risk strata (P < .01, respectively). Depressed men experienced worse overall mortality across risk strata (low: relative risk [RR], 1.86; 95% CI, 1.48 to 2.33; P < .001; intermediate: RR, 1.25; 95% CI, 1.06 to 1.49; P = .01; high: RR, 1.16; 95% CI, 1.03 to 1.32; P = .02). Men with intermediate- or high-risk prostate cancer and a recent diagnosis of depression are less likely to undergo definitive treatment and experience worse overall survival. The effect of depression disorders on prostate cancer treatment and survivorship warrants further study, because both conditions are relatively common in men in the United States.


An evaluation of bilirubin kinetics with respect to the diagnosis of Gilbert's syndrome.
1. The kinetics of the plasma disappearance of bilirubin (2 mg/kg intravenously) were studied in 106 patients with Gilbert's syndrome and in 13 normal subjects. 2. All patients had significant decreases in hepatic bilirubin clearance and transfer rates from plasma to liver, resulting in increased values for plasma retention at 4 h. The calculated value for unconugated bilirubin production was normal in 40% of patients and increased in the remainder. 3. In 29 of the Gilbert's patients their bromosulphthalein kinetics were studied 1 week before the bilirubin test. These results were essentially normal and it was concluded that the hepatic clearance mechanisms for bilirubin and bromosulphthalein are different. 4. In 10 patients the bilirubin transport maximum (Tm) was found to be low whereas the relative storage capacity (S) was normal. Phenobarbitone treatment in four patients resulted in an increase in Tm, and S decreased in two patients and remained unchanged in the other two. 5. These results support the hypothesis that there are several variants of Gilbert's syndrome and that the bilirubin tolerance test is a useful diagnostic test.


Optimal targeting of seasonal influenza vaccination toward younger ages is robust to parameter uncertainty.
Identification of the optimal vaccine allocation for the control of influenza requires consideration of uncertainty arising from numerous unpredictable factors, including viral evolution and diversity within the human population's immunity as well as variation in vaccine efficacy. The best policy must account for diverse potential outcomes based on these uncertainties. Here we used a mathematical model parametrized with survey-based contact data, demographic, and epidemiological data from seasonal influenza in the United States to determine the optimal vaccine allocation for five outcome measures: infections, hospitalizations, deaths, years of life loss, and contingent valuation. We incorporated uncertainty of epidemiological parameters and derive probability distributions of optimal age- and risk-specific allocation of vaccine. Our analysis demonstrated that previous recommendations of targeting schoolchildren (ages 5-17 years) and young adults (18-44 years) are generally robust in the face of uncertainty. However, when the outcome measure is to minimize deaths, years of life loss, or contingent valuation, uncertainty analysis identified scenarios under which it is optimal to target people at high risk for complications, even when vaccine are in abundance.


Fulfilment of knowledge expectations among family members of patients undergoing arthroplasty: a European perspective.
In the recovery process of arthroplasty patients, their family members play an important role due to short hospital stay and increased age of patients. Family members need to have knowledge to be able to support the patient. The aim of this study was to explore expected and received knowledge in family members of arthroplasty patients and describe the relationships between the differences in received and expected knowledge and background factors, country, information and control preferences and access to knowledge. The study was conducted in six European countries (Cyprus, Greece, Finland, Iceland, Spain and Sweden). The study design was cross-cultural, prospective and comparative with two measurement points: pre-operative and at discharge from hospital. Knowledge Expectations of significant other-scale and Krantz Health Opinion Survey were used before surgery and Received Knowledge of significant other-scale and Access to Knowledge at discharge. Patients undergoing elective hip or knee arthroplasty in seventeen hospitals were asked to identify one family member. The sample size was decided by power calculation. A total of 615 participants answered the questionnaires at both measurements. Family members perceived to receive less knowledge than they expected to have, most unfulfilled knowledge expectations were in the financial, social and experiential dimensions of knowledge. Seventy-four per cent of participants had unfulfilled knowledge expectations. Increased access to information from healthcare providers decreased the difference between received and expected knowledge. Compared to family members in southern Europe, those in the Nordic countries had more unfulfilled knowledge expectations and less access to information from healthcare providers. The evidence from this study highlights the need to involve the family members in the educational approach.


Insights into olfactory ensheathing cell development from a laser-microdissection and transcriptome-profiling approach.
Olfactory ensheathing cells (OECs) are neural crest-derived glia that ensheath bundles of olfactory axons from their peripheral origins in the olfactory epithelium to their central targets in the olfactory bulb. We took an unbiased laser microdissection and differential RNA-seq approach, validated by in situ hybridization, to identify candidate molecular mechanisms underlying mouse OEC development and differences with the neural crest-derived Schwann cells developing on other peripheral nerves. We identified 25 novel markers for developing OECs in the olfactory mucosa and/or the olfactory nerve layer surrounding the olfactory bulb, of which 15 were OEC-specific (that is, not expressed by Schwann cells). One pan-OEC-specific gene, Ptprz1, encodes a receptor-like tyrosine phosphatase that blocks oligodendrocyte differentiation. Mutant analysis suggests Ptprz1 may also act as a brake on OEC differentiation, and that its loss disrupts olfactory axon targeting. Overall, our results provide new insights into OEC development and the diversification of neural crest-derived glia.


Photomechanical movements of cultured embryonic photoreceptors: regulation by exogenous neuromodulators and by a regulable source of endogenous dopamine.
In the retina of nonmammalian vertebrates, light regulates photoreceptor morphology by causing rod photoreceptor elongation and cone photoreceptor contraction. The opposite photomechanical movements occur in the dark, and proceed with a circadian rhythm in many species in vivo. Using dissociated cultures of embryonic chick retina cells, we have recently demonstrated that photoreceptor cells that differentiate in vitro acquire the capacity of responding to light/dark cycles with photomechanical movements (Stenkamp and Adler, 1993). Here we report that the putative neuromodulators melatonin and dopamine can mimic the effects of darkness and light, respectively, on in vitro photomechanical movement. Pharmacological studies showed that dopamine appears to function by means of a D2-type receptor negatively coupled to adenylate cyclase. The effects of light on the cultured photoreceptors were inhibited by dopamine D2 receptor antagonists, and were attenuated by the dopaminergic neurotoxin 6-hydroxydopamine and by the dopamine synthesis inhibitor alpha-methyl-p-tyrosine. The possible existence of an endogenous source of dopamine in the cultures was also suggested by the presence of tyrosine hydroxylase-like immunoreactivity, and of an Na(+)-dependent mechanism for the accumulation of 3H-dopamine, which was predominantly associated with nonphotoreceptor cells. Additionally, 3H-dopamine release occurred in vitro through a Ca(2+)-dependent mechanism, as well as through reverse function of a nomifensine-sensitive dopamine transporter. Both of these putative release mechanisms appeared to be regulated by light and by melatonin, suggesting a mechanism whereby the putative dopaminergic cells may interact with other cells present in the cultures. These studies suggest that complex paracrine neuromodulatory mechanisms can differentiate in low-density embryonic cell culture, that dopaminergic activities exist in vitro, and that they are important for mediating photomechanical movements.


Home-based arm cycling exercise improves trunk control in persons with incomplete spinal cord injury: an observational study.
Arm cycling is used for cardiorespiratory rehabilitation but its therapeutic effects on the neural control of the trunk after spinal cord injury (SCI) remain unclear. We investigated the effects of single session of arm cycling on corticospinal excitability, and the feasibility of home-based arm cycling exercise training on volitional control of the erector spinae (ES) in individuals with incomplete SCI. Using transcranial magnetic stimulation, we assessed motor evoked potentials (MEPs) in the ES before and after 30 min of arm cycling in 15 individuals with SCI and 15 able-bodied controls (Experiment 1). Both groups showed increased ES MEP size after the arm cycling. The participants with SCI subsequently underwent a 6-week home-based arm cycling exercise training (Experiment 2). MEP amplitudes and activity of the ES, and movements of the trunk during reaching, self-initiated rapid shoulder flexion, and predicted external perturbation tasks were measured. After the training, individuals with SCI reached further and improved trajectory of the trunk during the rapid shoulder flexion task, accompanied by increased ES activity and MEP amplitudes. Exercise adherence was excellent. We demonstrate preserved corticospinal drive after a single arm cycling session and the effects of home-based arm cycling exercise training on trunk function in individuals with SCI.


Light-triggered modulation of cellular electrical activity by ruthenium diimine nanoswitches.
Ruthenium diimine complexes have previously been used to facilitate light-activated electron transfer in the study of redox metalloproteins. Excitation at 488 nm leads to a photoexcited state, in which the complex can either accept or donate an electron, respectively, in the presence of a soluble sacrificial reductant or oxidant. Here, we describe a novel application of these complexes in mediating light-induced changes in cellular electrical activity. We demonstrate that RubpyC17 ([Ru(bpy)(2)(bpy-C17)](2+), where bpy is 2,2'-bipyridine and bpy-C17 is 2,2'-4-heptadecyl-4'-methyl-bipyridine), readily incorporates into the plasma membrane of cells, as evidenced by membrane-confined luminescence. Excitable cells incubated in RubpyC17 and then illuminated at 488 nm in the presence of the reductant ascorbate undergo membrane depolarization leading to firing of action potentials. In contrast, the same experiment performed with the oxidant ferricyanide, instead of ascorbate, leads to hyperpolarization. These experiments suggest that illumination of membrane-associated RubpyC17 in the presence of ascorbate alters the cell membrane potential by increasing the negative charge on the outer face of the cell membrane capacitor, effectively depolarizing the cell membrane. We rule out two alternative explanations for light-induced membrane potential changes, using patch clamp experiments: (1) light-induced direct interaction of RubpyC17 with ion channels and (2) light-induced membrane perforation. We show that incorporation of RubpyC17 into the plasma membrane of neuroendocrine cells enables light-induced secretion as monitored by amperometry. While the present work is focused on ruthenium diimine complexes, the findings point more generally to broader application of other transition metal complexes to mediate light-induced biological changes.


Enzyme estimates of infarct size correlate with functional and clinical outcomes in the setting of ST-segment elevation myocardial infarction.
Cardiac biomarkers are routinely obtained in the setting of suspected myocardial ischemia and infarction. Evidence suggests these markers may correlate with functional and clinical outcomes, but the strength of this correlation is unclear. The relationship between enzyme measures of myocardial necrosis and left ventricular performance and adverse clinical outcomes were explored. Creatine kinase (CK) and CK-MB data were analyzed, as were left ventricular ejection fraction (LVEF) by angiogram, and infarct size by single-photon emission computed tomography (SPECT) imaging in patients in 2 trials: Prompt Reperfusion In Myocardial-infarction Evolution (PRIME), and Efegatran and Streptokinase to Canalize Arteries Like Accelerated Tissue plasminogen activator (ESCALAT). Both trials evaluated efegatran combined with thrombolysis for treating acute ST-segment elevation myocardial infarction (STEMI). Peak CK and CK area-under-the-curve (AUC) correlated significantly with SPECT-determined infarct size 5 to 10 days after enrollment. Peak CK had a statistically significant correlation with LVEF, but CK-AUC and LVEF correlation were less robust. Statistically significant correlations exist between SPECT-determined infarct size and peak CK-MB and CK-MB AUC. However, there was no correlation with LVEF for peak CK-MB and CK-MB AUC. The combined outcome of congestive heart failure and death were significantly associated with CK AUC, CK-MB AUC, peak CK, and peak CK-MB measurements. Peak CK and CK-MB values and AUC calculations have significant correlation with functional outcomes (LVEF- and SPECT-determined infarct size) and death or CHF outcomes in the setting of STEMI. Cardiac biomarkers provide prognostic information and may serve as valid endpoint measurements for phase II clinical trials.


Climate Change, Extreme Weather, and Intimate Partner Violence in East African Agrarian-Based Economies.
Severe weather events can be a catalyst for intimate partner violence, particularly in agricultural settings. This research explores the association between weather and violence in parts of East Africa that rely on subsistence farming. We used IPUMS-DHS data from Uganda in 2006, Zimbabwe in 2010, and Mozambique in 2011 for intimate partner violence frequency and EM-DAT data to identify weather events by region in the year of and year prior to IPUMS-DHS data collection. This work is grounded in a conceptual framework that illustrates the mechanisms through which violence increases. We used logistic regression to estimate the odds of reporting violence in regions with severe weather events. The odds of reporting violence were 25% greater in regions with severe weather compared to regions without in Uganda (OR = 1.25, 95% CI: 1.11-1.41), 38% greater in Zimbabwe (OR = 1.38, 95% CI: 1.13-1.70), and 91% greater in Mozambique (OR = 1.91, 95% CI: 1.64-2.23). Our results add to the growing body of evidence showing that extreme weather can increase women's and girls' vulnerability to violence. Moreover, this analysis demonstrates that climate justice and intimate partner violence must be addressed together.


Staphylococcal Immune Evasion Proteins: Structure, Function, and Host Adaptation.
Staphylococcus aureus is a successful human and animal pathogen. Its pathogenicity is linked to its ability to secrete a large amount of virulence factors. These secreted proteins interfere with many critical components of the immune system, both innate and adaptive, and hamper proper immune functioning. In recent years, numerous studies have been conducted in order to understand the molecular mechanism underlying the interaction of evasion molecules with the host immune system. Structural studies have fundamentally contributed to our understanding of the mechanisms of action of the individual factors. Furthermore, such studies revealed one of the most striking characteristics of the secreted immune evasion molecules: their conserved structure. Despite high-sequence variability, most immune evasion molecules belong to a small number of structural categories. Another remarkable characteristic is that S. aureus carries most of these virulence factors on mobile genetic elements (MGE) or ex-MGE in its accessory genome. Coevolution of pathogen and host has resulted in immune evasion molecules with a highly host-specific function and prevalence. In this review, we explore how these shared structures and genomic locations relate to function and host specificity. This is discussed in the context of therapeutic options for these immune evasion molecules in infectious as well as in inflammatory diseases.


Cell junctions in amphibian skin.
Cell junctions have been investigated in the amphibian epidermis, a stratified squamous epithelium, and compared to those described previously in simple columnar epithelia of mammalian cavitary organs. In adult frogs and toads, and in larvae approaching metamorphosis, belts of membrane fusion or zonulae occludentes of considerable depth are regularly found between adjoining cells of the outermost layer of the stratum corneum, binding the cells together into a continuous, uninterrupted sheet. Another set of occluding zonules appears in the second cornified layer (when such a layer is present), and a third set usually occurs in the outermost layer of the stratum granulosum. Specialized elements described as "modified" and "composite" desmosomes are encountered along the lateral and basal aspects, respectively, of the cornified cells; ordinary desmosomes and maculae occludentes (i.e., spots of membrane fusion) are found in all other strata. The usual 200 A intercellular gap is generally maintained between the cells of the stratum germinativum at the basal ends of the intercellular spaces. Hence, the intercellular spaces of the epidermis form a largely continuous network, closed to the external medium and open to the dermal interstitia. The situation is comparable to that found in columnar epithelia, except that the intercellular spaces are much more extensive, and an extracellular subcompartment (or two) apparently exists in the stratum corneum and between the latter and the stratum granulosum. The last subcompartment is usually filled with a dense substance, probably derived from discharged secretory granules. The tripartite junctional complex characteristic of lumen-lining epithelia (i.e., a zonula occludens followed by a zonula adhaerens, and desmosome) is seen only in early larvae; in adults and in larvae approaching metamorphosis, the occluding zonule is followed directly by a series of modified desmosomes. Interpreted in the light of current physiological data, these findings suggest that the diffusion of water, ions, and small, water-soluble molecules is impeded along the intercellular spaces of the epidermis by zonulae occludentes while it is facilitated from cell to cell within the epidermis by zonulae and maculae occludentes.


A Marked Point Process Framework for Extracellular Electrical Potentials.
Neuromodulations are an important component of extracellular electrical potentials (EEP), such as the Electroencephalogram (EEG), Electrocorticogram (ECoG) and Local Field Potentials (LFP). This spatially temporal organized multi-frequency transient (phasic) activity reflects the multiscale spatiotemporal synchronization of neuronal populations in response to external stimuli or internal physiological processes. We propose a novel generative statistical model of a single EEP channel, where the collected signal is regarded as the noisy addition of reoccurring, multi-frequency phasic events over time. One of the main advantages of the proposed framework is the exceptional temporal resolution in the time location of the EEP phasic events, e.g., up to the sampling period utilized in the data collection. Therefore, this allows for the first time a description of neuromodulation in EEPs as a Marked Point Process (MPP), represented by their amplitude, center frequency, duration, and time of occurrence. The generative model for the multi-frequency phasic events exploits sparseness and involves a shift-invariant implementation of the clustering technique known as k-means. The cost function incorporates a robust estimation component based on correntropy to mitigate the outliers caused by the inherent noise in the EEP. Lastly, the background EEP activity is explicitly modeled as the non-sparse component of the collected signal to further improve the delineation of the multi-frequency phasic events in time. The framework is validated using two publicly available datasets: the DREAMS sleep spindles database and one of the Brain-Computer Interface (BCI) competition datasets. The results achieve benchmark performance and provide novel quantitative descriptions based on power, event rates and timing in order to assess behavioral correlates beyond the classical power spectrum-based analysis. This opens the possibility for a unifying point process framework of multiscale brain activity where simultaneous recordings of EEP and the underlying single neuron spike activity can be integrated and regarded as marked and simple point processes, respectively.


Linking drugs to obscure illnesses: lessons from pure red cell aplasia, nephrogenic systemic fibrosis, and Reye's syndrome. a report from the Southern Network on Adverse Reactions (SONAR).
Identification of serious adverse drug reactions (sADRS) associated with commonly used drugs can elude detection for years. Reye's syndrome (RS), nephrogenic systemic fibrosis (NSF), and pure red cell aplasia (PRCA) among chronic kidney disease (CKD) patients were recognized in 1951, 2000, and 1998, respectively. Reports associating these syndromes with aspirin, gadodiamide, and epoetin, were published 29, 6, and 4 years later, respectively. We obtained primary information from clinicians who identified causes of these sADRs and reviewed factors contributing to delayed identification of these toxicities. Overall, 3,500 aspirin-associated RS cases in the United States, 1,605 gadolinium-associated NSF cases, and 181 epoetin-associated PRCA cases were reported. Delays in FDA regulation of over-the- counter medications and administration of aspirin to children contributed to development of RS. For NSF, in 1996, the Danish Medicine Agency approved high-dose gadodiamide administration to chronic kidney disease (CKD) patients undergoing MR scans. Overall, 88 % of Danish NSF cases were from two hospitals and 97 % of United States' NSF cases were from 60 hospitals. These hospitals frequently administered high-doses of gadodiamide to CKD patients. Another factor was the decision to administer linear chelated contrast agents versus lower risk macrocyclic chelated agents. For PRCA, increased use of subcutaneous epoetin formulations to CKD patients, in part due to convenience and cost-savings considerations, and a European regulatory requirement requiring removal of albumin as a stabilizer, led to toxicity. Overall, 81, 13, and 17 years elapsed between drug introduction into practice and identification of a causal relationship for aspirin, erythropoietin, and gadodiamide, respectively. A substantial decline in new cases of these sADRs occurred within two years of identification of the offending drug. Clinicians should be vigilant for sADRs, even for frequently-prescribed pharmaceuticals, particularly in settings where formulation or regulatory changes have occurred, or when over-the-counter, off-label, or pediatric use is common.