Structural basis of enzyme encapsulation into a bacterial nanocompartment.
Compartmentalization is an important organizational feature of life. It occurs at varying levels of complexity ranging from eukaryotic organelles and the bacterial microcompartments, to the molecular reaction chambers formed by enzyme assemblies. The structural basis of enzyme encapsulation in molecular compartments is poorly understood. Here we show, using X-ray crystallographic, biochemical and EM experiments, that a widespread family of conserved bacterial proteins, the linocin-like proteins, form large assemblies that function as a minimal compartment to package enzymes. We refer to this shell-forming protein as 'encapsulin'. The crystal structure of such a particle from Thermotoga maritima determined at 3.1-angstroms resolution reveals that 60 copies of the monomer assemble into a thin, icosahedral shell with a diameter of 240 angstroms. The interior of this nanocompartment is lined with conserved binding sites for short polypeptide tags present as C-terminal extensions of enzymes involved in oxidative-stress response.


Effects of frusemide on electrolyte and acid-base balance during exercise.
This study was undertaken to evaluate the effects of frusemide on the concentration of plasma electrolytes and the relationship between changes in electrolyte concentration and the simultaneous changes in acid-base balance in arterial and venous blood during intense exercise. Five exercise-conditioned Thoroughbred horses were exercised on a high-speed treadmill at a slope of 10% at speeds known to exceed VO2max. Horses participated in 3 randomised exercise trials in which they received either placebo (control), low-dose frusemide (0.5 mg/kg bwt), or high-dose frusemide (1.0 mg/kg) 4 h prior to exercise. Arterial and mixed venous blood samples were drawn before, during and after intense exercise. Packed cell volume (PCV), plasma protein, plasma electrolyte, plasma lactate concentrations and acid-base balance were determined. Frusemide at both dosages induced a significant metabolic alkalosis associated with a decrease in plasma çhloride[ in both arterial and mixed venous blood. There was a highly significant correlation between the strong ion difference (SID) and bicarbonate[ in all blood samples. The effects of frusemide on electrolyte concentration and acid-base balance were largely due to changes in SID.


Casein kinase II phosphorylation of the yeast phospholipid synthesis transcription factor Opi1p.
The transcription factor Opi1p regulates phospholipid synthesis in the yeast Saccharomyces cerevisiae by repressing the expression of several UAS(INO)-containing genes (e.g. INO1). Opi1p repressor activity is most active in inositol-supplemented cells. Regulation of Opi1p repressor activity is mediated by multiple phosphorylations catalyzed by protein kinases A and C. In this work, we showed that Opi1p was also phosphorylated by casein kinase II. Using purified maltose-binding protein-Opi1p as a substrate, casein kinase II activity was dose-and time-dependent and dependent on the concentrations of maltose-binding protein-Opi1p (Km = 25 microg/ml) and ATP (Km = 7 microM). Of three mutations (S10A, S38A, and S239A) in putative phosphorylation sites, 10 only the S10A mutation affected Opi1p phosphorylation. That Ser10 was a specific target of casein kinase II was confirmed by the loss of a phosphopeptide in the S10A mutant protein. The S10A mutation did not affect phosphorylation of Opi1p by either protein kinase A or protein kinase C. Likewise, phosphorylation of Opi1p by casein kinase II was not affected by mutations in protein kinase A (S31A and S251A) and protein (S10A) kinase C (S26A) phosphorylation sites. Expression of the OPI1 allele in an opi1Delta mutant attenuated (2-fold) the repressive effect of Opi1p on INO1 expression, and this effect was only observed when cells were grown in the absence of inositol. These data supported the conclusion that casein kinase II phosphorylation at Ser10 played a role in stimulating the repression of INO1 when Opi1p was not in its most active state (i.e. in inositol-deprived cells).


Glucose-sensitive QCM-sensors via direct surface RAFT polymerization.
Thin, phenylboronic acid-containing polymer coatings are potentially attractive sensory layers for a range of glucose monitoring systems. This contribution presents the synthesis and properties of glucose-sensitive polymer brushes obtained via surface RAFT polymerization of 3-methacrylamido phenylboronic acid (MAPBA). This synthetic strategy is attractive since it allows the controlled growth of PMAPBA brushes with film thicknesses of up to 20 nm via direct polymerization of MAPBA without the need for additional post-polymerization modification or deprotection steps. QCM-D sensor chips modified with a PMAPBA layer respond with a linear change in the shift of the fundamental resonance frequency over a range of physiologically relevant glucose concentrations and are insensitive toward the presence of fructose, thus validating the potential of these polymer brush films as glucose sensory thin coatings.


Prevalence of quadriceps muscle weakness in patients with COPD and its association with disease severity.
COPD presents with an array of extra-pulmonary symptoms of which skeletal muscle dysfunction, particularly of the quadriceps, is well recognized. This contributes to impaired quality of life and increased health care utilization. Work on the quadriceps originated from the observation that a good proportion of COPD patients stop exercise due to the feeling of leg fatigue rather than breathlessness. This study was carried out with the aim of finding the prevalence of quadriceps weakness in a population set and correlate it with severity of COPD. This cross-sectional study was conducted in 75 subjects suffering from COPD aged 45 years or above. COPD severity in the subjects was graded based on the GOLD staging system. A digital hand held dynamometer (HHD) was used to measure quadriceps muscle strength. Descriptive statistics were done, and Pearson's Correlation Coefficient and ANOVA analysis was used for expressing the results. Ninety two percent of subjects were suffering from quadriceps muscle weakness. Quadriceps weakness was present in significantly high proportions even in those suffering from mild disease and belonging to a younger age group. The mean quadriceps muscle force value decreased with disease severity and this relation was found to be significant (P<0.01). Majority of the COPD patients were found to be suffering from quadriceps weakness, which was also present in significant proportions in subjects belonging to younger age groups and suffering from mild disease. These findings indicate that onset of muscle weakness in COPD may precede the onset of symptoms. These findings suggest need for early remedial measure to prevent occurrence of associated systemic diseases.


Does racial concordance between HIV-positive patients and their physicians affect the time to receipt of protease inhibitors?
Compared to whites, African Americans have been found to have greater morbidity and mortality from HIV, partly due to their lower use of effective antiretroviral therapy. Why racial disparities in antiretroviral use exist is not completely understood. We examined whether racial concordance (patients and providers having the same race) affects the time of receipt of protease inhibitors. We analyzed data from a prospective, cohort study of a national probability sample of 1,241 adults receiving HIV care with linked data from 287 providers. We examined the association between patient-provider racial concordance and time from when the Food and Drug Administration approved the first protease inhibitor to the time when patients first received a protease inhibitor. In our unadjusted model, white patients received protease inhibitors much earlier than African-American patients (median 277 days compared to 439 days; P < .0001). Adjusting for patient characteristics only, African-American patients with white providers received protease inhibitors significantly later than African-American patients with African-American providers (median 461 days vs. 342 days respectively; P < .001) and white patients with white providers (median 461 vs. 353 days respectively; P= .002). In this model, no difference was found between African-American patients with African-American providers and white patients with white providers (342 vs. 353 days respectively; P > .20). Adjusting for patients' trust in providers, as well as other patient and provider characteristics in subsequent models, did not account for these differences. Patient-provider racial concordance was associated with time to receipt of protease inhibitor therapy for persons with HIV. Racial concordance should be addressed in programs, policies, and future racial and ethnic health disparity research.


Spry1 and Spry2 are necessary for eyelid closure.
Sproutys (Sprys) are downstream targets and negative feedback regulators of the FGF-Ras-ERK signaling pathway. Our previous studies have shown that Spry1 and Spry2, through negative modulation of FGF-ERK signaling, allow lens vesicle separation from the overlying ectoderm and regulate corneal epithelial proliferation. Here we show that Spry1 and Spry2 are necessary for eyelid closure. Murine palpebral conjunctival epithelial cells that differentiate as inner eyelids and adjacent mesenchymal cells express Spry1 and Spry2 prior to eyelid closure. Conditional deletion of both Spry1 and Spry2, but not either one alone, in the ocular surface epithelial cells result in the "EOB" (eyes open at birth) phenotype suggesting redundant roles for these proteins during eyelid closure. Spry mutant eyelids show increased proliferation of conjunctival epithelial cells with concomitant induction of FGF targets, Erm, Pea3 and Dusp6 and elevated ERK phosphorylation. Peridermal cells at the leading edge of Spry-mutant eyelids showed reduced c-Jun, but not ERK, phosphorylation, reduced F-actin polymerization and reduced motility in vitro. Spry mutant eyelids also showed disruptions in epithelial mesenchymal interactions reflected in the enhanced mesenchymal Spry1 and Spry4 expression, disaggregation of BMP4-positive mesenchymal cells and loss of Shh in the eyelid epithelium. Spry mutant eyelids also showed increased Wnt signaling and reduced expression of Foxc1 and Foxc2, two transcription factors previously shown to be necessary for eyelid closure. Collectively, our results show that conjunctival epithelial Spry1 and Spry2 redundantly promote eyelid closure by (a) stimulating ERK-independent, c-Jun-mediated peridermal migration, (b) suppressing conjunctival epithelial proliferation through FGF-ERK signaling, (c) mediating conjunctival epithelial-mesenchymal interactions and (d) maintaining expression of Foxc1 and Foxc2.


Correlation of structure, function and protein dynamics in GH7 cellobiohydrolases from Trichoderma atroviride, T. reesei and T. harzianum.
The ascomycete fungus Trichoderma reesei is the predominant source of enzymes for industrial conversion of lignocellulose. Its glycoside hydrolase family 7 cellobiohydrolase (GH7 CBH) TreCel7A constitutes nearly half of the enzyme cocktail by weight and is the major workhorse in the cellulose hydrolysis process. The orthologs from Trichoderma atroviride (TatCel7A) and Trichoderma harzianum (ThaCel7A) show high sequence identity with TreCel7A, ~ 80%, and represent naturally evolved combinations of cellulose-binding tunnel-enclosing loop motifs, which have been suggested to influence intrinsic cellobiohydrolase properties, such as endo-initiation, processivity, and off-rate. The TatCel7A, ThaCel7A, and TreCel7A enzymes were characterized for comparison of function. The catalytic domain of TatCel7A was crystallized, and two structures were determined: without ligand and with thio-cellotriose in the active site. Initial hydrolysis of bacterial cellulose was faster with TatCel7A than either ThaCel7A or TreCel7A. In synergistic saccharification of pretreated corn stover, both TatCel7A and ThaCel7A were more efficient than TreCel7A, although TatCel7A was more sensitive to thermal inactivation. Structural analyses and molecular dynamics (MD) simulations were performed to elucidate important structure/function correlations. Moreover, reverse conservation analysis (RCA) of sequence diversity revealed divergent regions of interest located outside the cellulose-binding tunnel of Trichoderma spp. GH7 CBHs. We hypothesize that the combination of loop motifs is the main determinant for the observed differences in Cel7A activity on cellulosic substrates. Fine-tuning of the loop flexibility appears to be an important evolutionary target in Trichoderma spp., a conclusion supported by the RCA data. Our results indicate that, for industrial use, it would be beneficial to combine loop motifs from TatCel7A with the thermostability features of TreCel7A. Furthermore, one region implicated in thermal unfolding is suggested as a primary target for protein engineering.


Soy food consumption does not lower LDL cholesterol in either equol or nonequol producers.
Health claims link soy protein (SP) consumption, through plasma cholesterol reduction, to a decreased risk of heart disease. Soy isoflavones (ISOs), particularly in individuals who produce equol, might also contribute to lipid lowering and thus reduce SP requirements. The objective was to examine the contributions of SP, ISOs, and equol to the hypocholesterolemic effects of soy foods. Nonsoy consumers (33 men, 58 women) with a plasma total cholesterol (TChol) concentration >5.5 mmol/L participated in a double-blind, placebo-controlled, crossover intervention trial. The subjects consumed 3 diets for 6 wk each in random order, which consisted of foods providing a daily dose of 1) 24 g SP and 70-80 mg ISOs (diet S); 2) 12 g SP, 12 g dairy protein (DP), and 70-80 mg ISOs (diet SD); and 3) 24 g DP without ISOs (diet D). Fasting plasma TChol, LDL cholesterol, HDL cholesterol, and triglycerides (TGs) were measured after each diet. TChol was 3% lower with the S diet (-0.17 +/- 0.06 mmol/L; P < 0.05) than with the D diet, and TGs were 4% lower with both the S (-0.14 +/- 0.05 mmol/L; P < 0.05) and SD (-0.12 +/- 0.05 mmol/L; P < 0.05) diets. There were no significant effects on LDL cholesterol, HDL cholesterol, or the TChol:HDL cholesterol ratio. On the basis of urinary ISOs, 30 subjects were equol producers. Lipids were not affected significantly by equol production. Regular consumption of foods providing 24 g SP/d from ISOs had no significant effect on plasma LDL cholesterol in mildly hypercholesterolemic subjects, regardless of equol-producing status.


Back injuries to fast bowlers in cricket: a prospective study.
Eighty-two high performance young male fast bowlers (mean age 16.8 years) were tested immediately prior to the season for selected kinanthropometric and physiological data. Subjects were also filmed both laterally (200 Hz) and from above (100 Hz) while bowling so that their front foot impacted a force platform during the delivery stride. The players then completed a log book over the ensuing season that detailed their training and playing programmes. All cricket related injuries over this season were assessed by a sports physician who used computerized tomography to assist in the diagnosis of spinal injuries. At the completion of this season the players were grouped according to their injury status (Group 1--bony injury to a vertebra; Group 2--soft tissue injury to the back that caused the player to miss at least one game, and Group 3--no injuries). A one-way analysis of variance was used to identify if any variables were significantly (P less than 0.05) different between the three groups, and a Scheffe post hoc comparison was used to determine which groups were significantly different. Eleven per cent of the players sustained a stress fracture to a vertebra(e) (L4 to S1), while 27 per cent sustained a soft tissue injury to the back. Bowlers with a low longitudinal foot arch were more likely to develop a stress fracture than those with a high arch. Shoulder depression and horizontal flexion strength for the preferred limb and quadriceps power in the non-preferred limb were also significantly related to back injuries. Results suggest that bowlers with the above physical characteristics, who bowl with these biomechanical techniques for extended periods, are predisposed to back injuries.


Dual modulation of MCL-1 and mTOR determines the response to sunitinib.
Most patients who initially respond to treatment with the multi-tyrosine kinase inhibitor sunitinib eventually relapse. Therefore, developing a deeper understanding of the contribution of sunitinib's numerous targets to the clinical response or to resistance is crucial. Here, we have shown that cancer cells respond to clinically relevant doses of sunitinib by enhancing the stability of the antiapoptotic protein MCL-1 and inducing mTORC1 signaling, thus evoking little cytotoxicity. Inhibition of MCL-1 or mTORC1 signaling sensitized cells to clinically relevant doses of sunitinib in vitro and was synergistic with sunitinib in impairing tumor growth in vivo, indicating that these responses are triggered as prosurvival mechanisms that enable cells to tolerate the cytotoxic effects of sunitinib. Furthermore, higher doses of sunitinib were cytotoxic, triggered a decline in MCL-1 levels, and inhibited mTORC1 signaling. Mechanistically, we determined that sunitinib modulates MCL-1 stability by affecting its proteasomal degradation. Dual modulation of MCL-1 stability at different dose ranges of sunitinib was due to differential effects on ERK and GSK3β activity, and the latter also accounted for dual modulation of mTORC1 activity. Finally, comparison of patient samples prior to and following sunitinib treatment suggested that increases in MCL-1 levels and mTORC1 activity correlate with resistance to sunitinib in patients.


Identification of single-copy orthologous genes between Physalis and Solanum lycopersicum and analysis of genetic diversity in Physalis using molecular markers.
The genus Physalis includes a number of commercially important edible and ornamental species. Its high nutritional value and potential medicinal properties leads to the increased commercial interest in the products of this genus worldwide. However, lack of molecular markers prevents the detailed study of genetics and phylogeny in Physalis, which limits the progress of breeding. In the present study, we compared the DNA sequences between Physalis and tomato, and attempted to analyze genetic diversity in Physalis using tomato markers. Blasting 23180 DNA sequences derived from Physalis against the International Tomato Annotation Group (ITAG) Release2.3 Predicted CDS (SL2.40) discovered 3356 single-copy orthologous genes between them. A total of 38 accessions from at least six species of Physalis were subjected to genetic diversity analysis using 97 tomato markers and 25 SSR markers derived from P. peruviana. Majority (73.2%) of tomato markers could amplify DNA fragments from at least one accession of Physalis. Diversity in Physalis at molecular level was also detected. The average Nei's genetic distance between accessions was 0.3806 with a range of 0.2865 to 0.7091. These results indicated Physalis and tomato had similarity at both molecular marker and DNA sequence levels. Therefore, the molecular markers developed in tomato can be used in genetic study in Physalis.


A Statistical Model of the Fluctuations in the Geomagnetic Field from Paleosecular Variation to Reversal
The statistical characteristics of the local magnetic field of Earth during paleosecular variation, excursions, and reversals are described on the basis of a database that gathers the cleaned mean direction and average remanent intensity of 2741 lava flows that have erupted over the last 20 million years. A model consisting of a normally distributed axial dipole component plus an independent isotropic set of vectors with a Maxwellian distribution that simulates secular variation fits the range of geomagnetic fluctuations, in terms of both direction and intensity. This result suggests that the magnitude of secular variation vectors is independent of the magnitude of Earth's axial dipole moment and that the amplitude of secular variation is unchanged during reversals.


Defining the "core microbiome" of the microbial communities in the tonsils of healthy pigs.
Porcine tonsils are the colonization site for many pathogenic as well as commensal microorganisms and are the primary lymphoid tissue encountered by organisms entering through the mouth or nares. The goal of this study was to provide an in-depth characterization of the composition and structure of the tonsillar microbial communities and to define the core microbiome in the tonsils of healthy pigs, using high throughput bar-coded 454-FLX pyrosequencing. Whole tonsils were collected at necropsy from 12 16-week-old finisher pigs from two healthy herds. Tonsil brushes were also used to collect samples from four of these animals. Bacterial DNA was isolated from each sample, amplified by PCR with universal primers specific for the bacterial 16S rRNA genes, and the PCR products sequenced using pyrosequencing. An average of 13,000 sequences were generated from each sample. Microbial community members were identified by sequence comparison to known bacterial 16S rRNA gene sequences.The microbiomes of these healthy herds showed very strong similarities in the major components as well as distinct differences in minor components. Pasteurellaceae dominated the tonsillar microbiome in all animals, comprising ~60% of the total, although the relative proportions of the genera Actinobacillus, Haemophilus, and Pasteurella varied between the herds. Also found in all animals were the genera Alkanindiges, Peptostreptococcus, Veillonella, Streptococcus and Fusobacterium, as well as Enterobacteriaceae and Neisseriaceae. Treponema and Chlamydia were unique to Herd 1, while Arcanobacterium was unique to Herd 2.Tonsil brushes yielded similar results to tissue specimens, although Enterobacteriaceae and obligate anaerobes were more frequently found in tissue than in brush samples, and Chlamydia, an obligately intracellular organism, was not found in brush specimens. We have extended and supported our previous studies with 16S clone libraries, using 16S rRNA gene pyrosequencing to describe the microbial communities in tonsils of healthy pigs. We have defined a core microbiome, dominated by Pasteurellaceae, in tonsil specimens, and have also demonstrated the presence of unique minor components of the tonsillar microbiome present in each herd. We have validated the use of non-invasive tonsil brushes, in comparison to tonsil tissue, which will facilitate future studies.


Development of a Cranberry Standard for Quantification of Insoluble Cranberry (Vaccinium macrocarpon Ait.) Proanthocyanidins.
Cranberry proanthocyanidins (PACs) can be partitioned into soluble PACs, which are extracted with solvents, and insoluble PACs, which remain associated with fibers and proteins after extraction. Most research on cranberry products only quantifies soluble PACs because proper standards for quantifying insoluble PACs are lacking. In this study, we evaluated the ability of a cranberry PAC (c-PAC) standard, reflective of the structural heterogeneity of PACs found in cranberry fruit, to quantify insoluble PACs by the butanol-hydrochloric acid (BuOH-HCl) method. For the first time, a c-PAC standard enabled conversion of BuOH-HCl absorbance values (550 nm) to a weight (milligram) basis, allowing for quantification of insoluble PACs in cranberries. The use of the c-PAC reference standard for sequential analysis of soluble PACs by the method of 4-(dimethylamino)cinnamaldehyde and insoluble PACs by the method of BuOH-HCl provides analytical tools for the standardization of cranberry-based ingredients.


Biogenesis of mitochondria: dual role of Tom7 in modulating assembly of the preprotein translocase of the outer membrane.
Biogenesis of the translocase of the outer mitochondrial membrane (TOM complex) involves the assembly of the central β-barrel forming protein Tom40 with six different subunits that are embedded in the membrane via α-helical transmembrane segments. The sorting and assembly machinery (SAM complex) of the outer membrane plays a central role in this process. The SAM complex mediates the membrane integration of β-barrel precursor proteins including Tom40. The small Tom proteins Tom5 and Tom6 associate with the precursor of Tom40 at the SAM complex at an early stage of the assembly process and play a stimulatory role in the formation of the mature TOM complex. A fraction of the SAM components interacts with the outer membrane protein mitochondrial distribution and morphology protein 10 (Mdm10) to form the SAM-Mdm10 machinery; however, different views exist on the function of the SAM-Mdm10 complex. We report here that the third small Tom protein, Tom7, plays an inhibitory role at two distinct steps in the biogenesis of the TOM complex. First, Tom7 plays an antagonistic role to Tom5 and Tom6 at the early stage of Tom40 assembly at the SAM complex. Second, Tom7 interacts with Mdm10 that is not bound to the SAM complex, and thus promotes dissociation of the SAM-Mdm10 complex. Since the SAM-Mdm10 complex is required for the biogenesis of Tom22, Tom7 delays the assembly of Tom22 with Tom40 at a late stage of assembly of the TOM complex. Thus, Tom7 modulates the biogenesis of topologically different proteins, the β-barrel forming protein Tom40 and Tom22 that contains a transmembrane α-helix.


Comparisons of bidirectional reflectance distribution function measurements on prepared particulate surfaces and radiative-transfer models.
To understand the connection between single-particle optics and the optics of a closely packed surface, controlled laboratory measurements of bidirectional reflectance distribution functions on layers of polymer and glass spheres are carried out. The measurements are compared with predictions from five radiative-transfer models; the Hapke's models, the Lumme-Bowell model, the BRF algorithm of Mishchenko et al., and the discrete ordinate radiative transfer. It is found that models of strict numerical radiative-transfer equations (RTEs) predict measurements well in some regions but have errors in both forward- and backward-scattering directions. The improved Hapke's model, although it has an anisotropic multiple-scattering term, still produces considerable errors compared with the strict RTE. The difference can be attributed to the exclusion of a diffraction contribution in the Hapke model.


HIV-associated neurocognitive disorders before and during the era of combination antiretroviral therapy: differences in rates, nature, and predictors.
Combination antiretroviral therapy (CART) has greatly reduced medical morbidity and mortality with HIV infection, but high rates of HIV-associated neurocognitive disorders (HAND) continue to be reported. Because large HIV-infected (HIV+) and uninfected (HIV-) groups have not been studied with similar methods in the pre-CART and CART eras, it is unclear whether CART has changed the prevalence, nature, and clinical correlates of HAND. We used comparable methods of subject screening and assessments to classify neurocognitive impairment (NCI) in large groups of HIV + and HIV - participants from the pre-CART era (1988-1995; N = 857) and CART era (2000-2007; N = 937). Impairment rate increased with successive disease stages (CDC stages A, B, and C) in both eras: 25%, 42%, and 52% in pre-CART era and 36%, 40%, and 45% in CART era. In the medically asymptomatic stage (CDC-A), NCI was significantly more common in the CART era. Low nadir CD4 predicted NCI in both eras, whereas degree of current immunosuppression, estimated duration of infection, and viral suppression in CSF (on treatment) were related to impairment only pre-CART. Pattern of NCI also differed: pre-CART had more impairment in motor skills, cognitive speed, and verbal fluency, whereas CART era involved more memory (learning) and executive function impairment. High rates of mild NCI persist at all stages of HIV infection, despite improved viral suppression and immune reconstitution with CART. The consistent association of NCI with nadir CD4 across eras suggests that earlier treatment to prevent severe immunosuppression may also help prevent HAND. Clinical trials targeting HAND prevention should specifically examine timing of ART initiation.


A sequence downstream of AAUAAA is required for rabbit beta-globin mRNA 3'-end formation.
The sequence AAUAAA, found 11-30 base pairs (bp) upstream of the poly(A) site of most non-histone eukaryotic messenger RNAs (mRNAs) forms an essential part of the recognition site for 3'-end processing of the primary transcript. However, the sequence AATAAA is found in transcribed regions of genes and is differentially utilized in genes containing multiple copies of the sequence within the 3'-noncoding region, suggesting that the hexanucleotide alone does not comprise a complete recognition site. Therefore, it seems likely that additional sequences are required to form a complete recognition site for 3'-end formation. We have investigated the sequence requirements for mRNA 3'-end formation using the rabbit beta-globin gene as a model system. Here we demonstrate that an additional sequence 3' to AAUAAA is required for the correct 3'-end formation of rabbit beta-globin mRNA.


Considerations for hydrogel applications to neural bioelectronics.
Hydrogels have been applied across a wide range of biomedical applications due to their versatility, but more recently have garnered interest as materials in bioelectronics due to the capacity to tailor their mechanical and biological properties. Hydrogel coatings in particular have been used to impart softness at the bionic device interface, deliver therapeutics and control cell interactions through presentation of peptides and growth factors. Additionally, the use of dynamic hydrogel properties has been harnessed as shuttles for the implantation of flexible electrode arrays. In all of these applications, the hydrogel must be designed not only to provide the desired performance, but also have no unexpected impacts on the surrounding tissues, such as extensive swelling that can compress the cells at the interface. Appropriate selection and design of hydrogel systems for bioelectronics requires an understanding of the physical, chemical and biological properties of hydrogels as well as their structure-property relationships. This review covers the design rationale for application of hydrogels systems for use in bioelectronic devices with a focus on in vivo applications.


Smelling a single component of male sweat alters levels of cortisol in women.
Rodents use chemosignals to alter endocrine balance in conspecifics. Although responses to human sweat suggest a similar mechanism in humans, no particular component of human sweat capable of altering endocrine balance in conspecifics has yet been isolated and identified. Here, we measured salivary levels of the hormone cortisol in women after smelling pure androstadienone (4,16-androstadien-3-one), a molecule present in the sweat of men that has been suggested as a chemosignal in humans. We found that merely smelling androstadienone maintained significantly higher levels of the hormone cortisol in women. These results suggest that, like rodents, humans can influence the hormonal balance of conspecifics through chemosignals. Critically, this study identified a single component of sweat, androstadienone, as capable of exerting such influence. This result points to a potential role for synthetic human chemosignals in clinical applications.


Antibody and T-cell responses associated with experimental human malaria infection or vaccination show limited relationships.
This study examined specific antibody and T-cell responses associated with experimental malaria infection or malaria vaccination, in malaria-naive human volunteers within phase I/IIa vaccine trials, with a view to investigating inter-relationships between these types of response. Malaria infection was via five bites of Plasmodium falciparum-infected mosquitoes, with individuals reaching patent infection by 11-12 days, having harboured four or five blood-stage cycles before drug clearance. Infection elicited a robust antibody response against merozoite surface protein-119 , correlating with parasite load. Classical class switching was seen from an early IgM to an IgG1-dominant response of increasing affinity. Malaria-specific T-cell responses were detected in the form of interferon-γ and interleukin-4 (IL-4) ELIspot, but their magnitude did not correlate with the magnitude of antibody or its avidity, or with parasite load. Different individuals who were immunized with a virosome vaccine comprising influenza antigens combined with P. falciparum antigens, demonstrated pre-existing interferon-γ, IL-2 and IL-5 ELIspot responses against the influenza antigens, and showed boosting of anti-influenza T-cell responses only for IL-5. The large IgG1-dominated anti-parasite responses showed limited correlation with T-cell responses for magnitude or avidity, both parameters being only negatively correlated for IL-5 secretion versus anti-apical membrane antigen-1 antibody titres. Overall, these findings suggest that cognate T-cell responses across a range of magnitudes contribute towards driving potentially effective antibody responses in infection-induced and vaccine-induced immunity against malaria, and their existence during immunization is beneficial, but magnitudes are mostly not inter-related.


Guidelines for the Detection of Babesia and Theileria Parasites.
The genera Babesia and Theileria (phylum Apicomplexa, order Piroplasmida) are mainly transmitted by Ixodid ticks in which the sexual part of their life cycle followed by sporogony takes place. They include protozoan parasites that infect erythrocytes of a variety of vertebrate hosts, including domestic and wild animals, with some Babesia spp. also infecting humans. Babesia sporozoites transmitted in the tick's saliva during the bloodmeal directly infect erythrocytes, where they asexually multiply to produce pear-shaped merozoites in the process of merogony; whereas a pre-erythrocytic schizogonic life stage in leukocytes is found in Theileria and precedes merogony in the erythrocytes. The wide spectrum of Babesia and Theileria species and their dissimilar characteristics with relation to disease severity, transmission, epidemiology, and drug susceptibility stress the importance of accurate detection of babesiosis and theileriosis and their causative agents. These guidelines review the main methods currently used for the detection of Babesia and Theileria spp. for diagnostic purposes as well as epidemiological studies involving their vertebrate hosts and arthropod vectors. Serological methods were not included once they did not indicate current infection but rather exposure.


Quantifying the Nucleation and Growth Kinetics of Microwave Nanochemistry Enabled by in Situ High-Energy X-ray Scattering.
The fast reaction kinetics presented in the microwave synthesis of colloidal silver nanoparticles was quantitatively studied, for the first time, by integrating a microwave reactor with in situ X-ray diffraction at a high-energy synchrotron beamline. Comprehensive data analysis reveals two different types of reaction kinetics corresponding to the nucleation and growth of the Ag nanoparticles. The formation of seeds (nucleation) follows typical first-order reaction kinetics with activation energy of 20.34 kJ/mol, while the growth of seeds (growth) follows typical self-catalytic reaction kinetics. Varying the synthesis conditions indicates that the microwave colloidal chemistry is independent of concentration of surfactant. These discoveries reveal that the microwave synthesis of Ag nanoparticles proceeds with reaction kinetics significantly different from the synthesis present in conventional oil bath heating. The in situ X-ray diffraction technique reported in this work is promising to enable further understanding of crystalline nanomaterials formed through microwave synthesis.


Strategic Interviewing to Detect Deception: Cues to Deception across Repeated Interviews.
Previous deception research on repeated interviews found that liars are not less consistent than truth tellers, presumably because liars use a "repeat strategy" to be consistent across interviews. The goal of this study was to design an interview procedure to overcome this strategy. Innocent participants (truth tellers) and guilty participants (liars) had to convince an interviewer that they had performed several innocent activities rather than committing a mock crime. The interview focused on the innocent activities (alibi), contained specific central and peripheral questions, and was repeated after 1 week without forewarning. Cognitive load was increased by asking participants to reply quickly. The liars' answers in replying to both central and peripheral questions were significantly less accurate, less consistent, and more evasive than the truth tellers' answers. Logistic regression analyses yielded classification rates ranging from around 70% (with consistency as the predictor variable), 85% (with evasive answers as the predictor variable), to over 90% (with an improved measure of consistency that incorporated evasive answers as the predictor variable, as well as with response accuracy as the predictor variable). These classification rates were higher than the interviewers' accuracy rate (54%).


Anxiety and depressive symptoms are associated with poor sleep health during a period of COVID-19-induced nationwide lockdown: a cross-sectional analysis of adults in Jordan.
Jordan, a Middle Eastern country, declared a state of national emergency due to COVID-19 and a strict nationwide lockdown on 17 March 2020, banning all travel and movement around the country, potentially impacting mental health. This study sought to investigate the association between mental health (eg, anxiety and depressive symptoms) and sleep health among a sample of Jordanians living through a state of COVID-19-induced nationwide lockdown. Using Facebook, participants (n=1240) in Jordan in March 2020 were recruited and direct to a web-based survey measuring anxiety (items from General Anxiety Disorder 7-item (GAD-7) scale instrument), depressive symptoms (items from Center for Epidemiologic Studies Depression Scale), sleep health (items from the Pittsburgh Sleep Quality Index) and sociodemographic. A modified Poisson regression model with robust error variance. Adjusted prevalence ratios (aPRs) and 95% CIs were estimated to examine how anxiety and depressive symptoms may affect different dimensions of sleep health: (1) poor sleep quality, (2) short sleep duration, (3) encountering sleep problems. The majority of participants reported having experienced mild (33.8%), moderate (12.9%) or severe (6.3%) levels of anxiety during lockdown, and nearly half of respondents reported depressive symptoms during lockdown. Similarly, over 60% of participants reported having experienced at least one sleep problem in the last week, and nearly half reported having had short sleep duration. Importantly, anxiety was associated with poor sleep health outcomes. For example, corresponding to the dose-response relationship between anxiety and sleep health outcomes, those reporting severe anxiety were the most likely to experience poor sleep quality (aPR =8.95; 95% CI=6.12 to 13.08), short sleep duration (aPR =2.23; 95% CI=1.91 to 2.61) and at least one problem sleep problem (aPR=1.73; 95% CI=1.54 to 1.95). Moreover, depressive symptoms were also associated with poor sleep health outcomes. As compared with scoring in the first quartile, scoring fourth quartile was associated with poor sleep quality (aPR=11.82; 95% CI=6.64 to 21.04), short sleep duration (aPR=1.87; 95% CI=1.58 to 2.22), and experiencing at least one sleep problem (aPR=1.90; 95% CI=1.66 to 2.18). Increased levels of anxiety and depressive symptoms can negatively influence sleep health among a sample of Jordanian adults living in a state of COVID-19-induced nationwide lockdown.


Intra-abdominal fat. Part II: Non-cancerous lesions of the adipose tissue localized beyond organs.
Adipose tissue does not belong to the most favorite structures to be visualized by ultrasound. It is not, however, free from various pathologies. The aim of this paper is to make abdominal cavity examiners more familiar with non-cancerous lesions found in intra-abdominal fat. The main focus is lesions that are rarely discussed in the literature. Visceral adiposity is one of important pathogenetic factors contributing to cardiovascular events, metabolic syndrome and even certain neoplasms. That is why this article exposes sonographic features that are the most characteristic of these lesions. The value of ultrasonography in the diagnosis of this pathology is underestimated, and a number of US scan reports do not reflect its presence in any way. Moreover, the article discusses more and more common mesenteritis, the lack of knowledge of which could pose difficulties in explaining the nature of symptoms reported by patients. Furthermore, this review presents lesions referred to in the literature as focal infarction of intra-abdominal fat. This section focuses on infarction of the greater and lesser omentum, epiploic appendagitis, mesenteric volvulus and focal fat necrosis resulting from pancreatitis. These lesions should be assessed with respect to the clinical context, and appropriate techniques of ultrasonography should be employed to allow careful determination of the size, shape, acoustic nature and location of lesions in relation to the integuments and large bowel, as well as their reaction to compression with an ultrasound transducer and behavior during deep inspiration. Moreover, each lesion must be obligatorily assessed in terms of blood flow. Doppler evaluation enables the differentiation between primary and secondary inflammation of intra-abdominal fat. The paper also draws attention to a frequent indirect sign of a pathological process, i.e. thickening and hyperechogenicity of fat, which sometimes indicates an ongoing pathology at a deeper site. This structure may completely conceal the primary lesion rendering it inaccessible for ultrasound. In such cases and in the event of other doubts, computed tomography should be the next diagnostic step.


Blind deconvolution of images using optimal sparse representations.
The relative Newton algorithm, previously proposed for quasi-maximum likelihood blind source separation and blind deconvolution of one-dimensional signals is generalized for blind deconvolution of images. Smooth approximation of the absolute value is used as the nonlinear term for sparse sources. In addition, we propose a method of sparsification, which allows blind deconvolution of arbitrary sources, and show how to find optimal sparsifying transformations by supervised learning.


Multimorbidity in rheumatic conditions.
In recent years, multimorbidity in rheumatic conditions has gained increasing attention. Rheumatologist care for an aging patient population with multiple diseases, therefore multimorbidity is the rule, not the exception. Owing to the high prevalence and the potential interaction of coexisting diseases, multimorbidity needs to be taken into account when treating patients with chronic inflammatory conditions. In this review we address the most prevalent comorbidities in patients with rheumatic conditions and their impact on important outcomes, such as physical function, quality of life, and mortality.


Subtelomeric Rearrangements: Presentation of 21 Probands with Emphasis on Familial Cases.
Intellectual disability affects 2% - 3% of the general population, with a chromosomal abnormality being found in 4% - 28% of these patients and a cryptic subtelomeric abnormality in 3% - 16%. In most cases, these subtelomeric rearrangements are submicroscopic, requiring techniques other than conventional karyotype for detection. They may be de novo or inherited from an affected parent or from a healthy carrier of a balanced chromosomal abnormality. The aim of this study was to characterize patients from our medical genetics center, in whom both a deletion and duplication in subtelomeric regions were found. Clinical and cytogenetic characterization of 21 probands followed at our center, from 1998 until 2017, with subtelomeric rearrangements. There were 21 probands from 19 families presenting with intellectual disability and facial dysmorphisms. Seven had behavior changes, five had epilepsy and 14 presented with some other sign or symptom. Four had chromosomal abnormalities detected by conventional karyotype and four were diagnosed by array-comparative genomic hybridization. In four cases, parental studies were not possible. The online mendelian inheritance in man classification was provided whenever any of the phenotypes (deletion or duplication syndrome) was dominant. Patients and relevant family members were clinically and cytogenetically characterized. Although rare, subtelomeric changes are a substantial cause of syndromic intellectual disability with important familial repercussions. It is essential to remember that a normal array-comparative genomic hybridization result does not exclude a balanced rearrangement in the parents. Parental genetic studies are essential not only for a complete characterization of the rearrangement, but also for accurate genetic counselling and screening of family members at risk for recurrence.


Four-year metabolic outcome of a randomised controlled CD3-antibody trial in recent-onset type 1 diabetic patients depends on their age and baseline residual beta cell mass.
The aim of the study was to examine the 48 month outcome of treating recent-onset type 1 diabetic patients for 6 days with humanised CD3-antibody, ChAglyCD3. Eighty patients, aged 12-39 years, were recruited for a phase 2 multicentre trial and randomised to placebo (n=40) or ChAglyCD3 (n=40) treatment by a third party member; participants and care-givers were blinded. The change in insulin dose (U kg(-1)day(-1)) over 48 months was chosen as primary endpoint and compared in 31 placebo-and 33 ChAglyCD3-treated patients. Adverse events were followed in 35 and 38 patients, respectively. Treatment with ChAglyCD3 delayed the rise in insulin requirements of patients with recent-onset diabetes and reduced its amplitude over 48 months (+0.09 vs +0.32 U kg(-1)day(-1) in the placebo group). Using multivariate analysis this effect was correlated with higher baseline residual beta cell function and a younger age. It was associated with better outcome variables in subgroups selected according to these variables. In the ChAglyCD3 subgroup with higher initial beta cell function, 0/11 patients became C-peptide-negative over 48 months vs 4/9 in the corresponding placebo subgroup. In the subgroup aged <27 years old, antibody treatment preserved initial beta cell function for 36 months (vs >80% decline within 24 months in the placebo subgroup <27 years old), resulted in lower HbA1c concentrations and tended to reduce glycaemic variability (p=0.08). No longterm adverse events were observed. A 6 day ChAglyCD3 treatment can suppress the rise in insulin requirements of recent-onset type 1 diabetic patients over 48 months, depending on their age and initial residual beta cell function. In younger patients this effect is associated with reduced deterioration of metabolic variables. These observations help to define inclusion criteria for prevention trials. ClinicalTrials.gov NCT00627146 Center grants from the Juvenile Diabetes Research Foundation (4-2001-434, 4-2005-1327) and grants from the Belgian Fund for Scientific Research-Flanders and from Brussels Free University-VUB.


A Novel Triage Approach to Identifying Health Concerns.
We investigated the detection of health problems in preventive child health care (PCH) by a novel triage approach for routine health assessments. In the triage approach, all children were preassessed by a physician's assistant, and only those in need of follow-up were assessed by a PCH physician or nurse. In the traditional approach, all children were assessed by a PCH physician or nurse. A prospective cohort design was used with data on routine assessments of 1897 children aged 5 to 6 and 10 to 11 years. Primary outcomes were the detection of overweight, visual disorders, and psychosocial problems, with type of approach (traditional vs triage) as independent variable. To assess the severity of health problems, BMI, Snellen, Strengths and Difficulties Questionnaire, and Child Behavior Checklist, scores were compared for both approaches in subgroups of children with overweight, visual disorders, or psychosocial problems. No significant differences were found between the approaches in terms of the detection of incident cases of overweight, visual disorders, and psychosocial problems. Significantly higher Strengths and Difficulties Questionnaire scores were found in the subgroup with psychosocial problems when the triage approach was used. Marginal differences between the approaches were found for severity of overweight in the subgroup of overweight children. A novel triage approach to PCH resulting in less involvement of physicians and nurses in routine assessments appears to detect health problems as effectively as the traditional approach in place. More research is needed to determine the long-term outcomes of the 2 approaches.


Analysis of Health-Care Charges in Murine Typhus: Need for Improved Clinical Recognition and Diagnostics for Acute Disease.
Murine typhus, caused by Rickettsia typhi, is an undifferentiated febrile illness with no available rapid and sensitive diagnostic assay for use during early disease. We aimed to compare the health-care charges in those diagnosed with murine typhus to those with influenza, a febrile illness with an available rapid diagnostic test. A comparison of health-care-associated charges at the University of Texas Medical Branch at Galveston demonstrated a median of $817 for influenza versus $16,760 for murine typhus (P < 0.0001). Median laboratory ($184 versus $3,254 [P < 0.0001]) and imaging charges ($0 versus $514 [P < 0.0001]) were also higher in those with murine typhus. Those receiving at least one imaging study during their illness were greater in the murine typhus group (91.3% versus 20.3%) (P < 0.0001). The median time needed to establish a confirmed or presumptive diagnosis was 2 days for influenza compared with 9 days for murine typhus (P < 0.0001). The median number of health-care encounters was greater for those with murine typhus (2 versus 1) (P < 0.0001). Eleven patients (15.9%) with influenza were hospitalized as a result of their illness compared with 16 (69.6%) with murine typhus (P < 0.0001). The estimated mortality based on disease severity at presentation by Acute Physiology and Chronic Health Evaluation II scoring was similar in the two groups-both had a median 4% mortality risk (P = 0.0893). These results highlight the need for improved clinical recognition and diagnostics for acute rickettsioses such as murine typhus.


Directed evolution generates a novel oncolytic virus for the treatment of colon cancer.
Viral-mediated oncolysis is a novel cancer therapeutic approach with the potential to be more effective and less toxic than current therapies due to the agents selective growth and amplification in tumor cells. To date, these agents have been highly safe in patients but have generally fallen short of their expected therapeutic value as monotherapies. Consequently, new approaches to generating highly potent oncolytic viruses are needed. To address this need, we developed a new method that we term "Directed Evolution" for creating highly potent oncolytic viruses. Taking the "Directed Evolution" approach, viral diversity was increased by pooling an array of serotypes, then passaging the pools under conditions that invite recombination between serotypes. These highly diverse viral pools were then placed under stringent directed selection to generate and identify highly potent agents. ColoAd1, a complex Ad3/Ad11p chimeric virus, was the initial oncolytic virus derived by this novel methodology. ColoAd1, the first described non-Ad5-based oncolytic Ad, is 2-3 logs more potent and selective than the parent serotypes or the most clinically advanced oncolytic Ad, ONYX-015, in vitro. ColoAd1's efficacy was further tested in vivo in a colon cancer liver metastasis xenograft model following intravenous injection and its ex vivo selectivity was demonstrated on surgically-derived human colorectal tumor tissues. Lastly, we demonstrated the ability to arm ColoAd1 with an exogenous gene establishing the potential to impact the treatment of cancer on multiple levels from a single agent. Using the "Directed Evolution" methodology, we have generated ColoAd1, a novel chimeric oncolytic virus. In vitro, this virus demonstrated a >2 log increase in both potency and selectivity when compared to ONYX-015 on colon cancer cells. These results were further supported by in vivo and ex vivo studies. Furthermore, these results have validated this methodology as a new general approach for deriving clinically-relevant, highly potent anti-cancer virotherapies.


Consulting the oracle: ten lessons from using the Delphi technique in nursing research.
The aim of this paper was to provide insight into the Delphi technique by outlining our personal experiences during its use over a 10-year period in a variety of applications. As a means of achieving consensus on an issue, the Delphi research method has become widely used in healthcare research generally and nursing research in particular. The literature on this technique is expanding, mainly addressing what it is and how it should be used. However, there is still much confusion and uncertainty surrounding it, particularly about issues such as modifications, consensus, anonymity, definition of experts, how 'experts' are selected and how non-respondents are pursued. This issues that arise when planning and carrying out a Delphi study include the definition of consensus; the issue of anonymity vs. quasi-anonymity for participants; how to estimate the time needed to collect the data, analyse each 'round', feed back results to participants, and gain their responses to this feedback; how to define and select the 'experts' who will be asked to participate; how to enhance response rates; and how many 'rounds' to conduct. Many challenges and questions are raised when using the Delphi technique, but there is no doubt that it is an important method for achieving consensus on issues where none previously existed. Researchers need to adapt the method to suit their particular study.


Occurrence of antimicrobial residues in Brazilian food animals in 2008 and 2009.
Brazil is one of the most important countries as a producer and exporter of cattle and poultry. In 2009 cattle accounted for 30% of the export market and 41.4% for poultry meat. The Brazilian National Residues and Contaminants Control Plan (PNCRC) follows the guidelines set by the Codex Alimentarius Commission and checks compliance maximum residue limits (MRLs) to ensure the quality of these commodities. Kidney samples (n = 2978) were analysed between January 2008 and December 2009. Fifteen antibiotics of the macrolide and aminoglycoside groups (clindamycin, eritromycin, lincomycin, tylmicosin, tylosin, amikacin, apramycin, dihydrostreptomycin, gentamycin, higromycin, kanamycin, neomycin, spectinomycin, streptomycin, tobramycin) were determined by a microbiological screening method (FAST) and confirmed/quantified using liquid chromatography (LC-MS/MS and UPLC-MS/MS). In 2008, 1459 samples were analysed by a screening test and liquid chromatography with only one sample (0.07%) exceeded Brazilian legislation limits (>MRL). In 2009, 1519 samples were analysed and none exceeding Brazilian legislation limits (>MRL). The slaughterhouses of 16 states were monitored during the year of 2008, and 18 states were monitored in 2009, being the major producing states most sampled by the PNCRC.


Effect of 12 months of whole-body vibration therapy on bone density and structure in postmenopausal women: a randomized trial.
Although data from studies in animals demonstrated beneficial effects of whole-body vibration (WBV) therapy on bone, clinical trials in postmenopausal women showed conflicting results. To determine whether WBV improves bone density and structure. A 12-month, single-center, superiority, randomized, controlled trial with 3 parallel groups. (ClinicalTrials.gov registration number: NCT00420940) Toronto General Hospital, Ontario, Canada. 202 healthy postmenopausal women with bone mineral density (BMD) T-scores between -1.0 and -2.5 who were not receiving prescription bone medications. Participants were randomly assigned to 1 of 3 groups (1:1:1 ratio) by using a block-randomization scheme and sealed envelopes. They were asked to stand on a low-magnitude (0.3g) 90-Hz or 30-Hz WBV platform for 20 minutes daily or to serve as control participants; all participants received calcium and vitamin D. Bone outcome assessors, who were blinded to group assignment, determined trabecular volumetric BMD and other measurements of the distal tibia and distal radius with high-resolution peripheral quantitative computed tomography and areal BMD with dual-energy x-ray absorptiometry at baseline and at 12 months. 12 months of WBV therapy had no significant effect on any bone outcomes compared with no WBV therapy. For the primary outcome of tibial trabecular volumetric BMD, mean change from baseline was 0.4 mg/cm(3) (95% CI, -0.4 to 1.2 mg/cm(3)) in the 90-Hz WBV group, -0.1 mg/cm(3) (CI, -1.0 to 0.8 mg/cm(3)) in the 30-Hz WBV group, and -0.2 mg/cm(3) (CI, -1.1 to 0.6 mg/cm(3)) in the control group (P = 0.55). Changes in areal BMD at the femoral neck, total hip, and lumbar spine were also similar among the groups. Overall, low-magnitude WBV at both 90 and 30 Hz was well-tolerated. Adherence to WBV ranged from 65% to 79%. Double-blinding was not possible. Whole-body vibration therapy at 0.3g and 90 or 30 Hz for 12 months did not alter BMD or bone structure in postmenopausal women who received calcium and vitamin D supplementation.


Novel imaging approaches to head and neck cancer.
An inadequate supply of oxygen, hypoxia, is an important factor contributing to resistance to treatment in a number of tumor types, including head and neck cancer. Novel imaging methods have been applied to studies of this important prognostic factor. Mammalian cells need oxygen to live but O2 also participates in the cytotoxic effects of ionizing radiation. Hypoxia is often the result of abnormal blood vessels supplying the tumor, increased diffusion distances to tumor cells, and reduced O2 transport capacity of the blood. Its consequences are mediated by a series of hypoxia-initiated genomic changes activating angiogenesis, glycolysis, and other processes that enable tumor cells to survive or escape the O2-deficient environment. Hypoxia has been shown to be important in overall diminished therapeutic response, malignant progression, increased probability of recurrence, locoregional spread, and distant metastases. Strategies are being developed to surmount the cure-limiting consequences of hypoxia, but methods are needed to select patients most likely to benefit from these new treatments. Even though hypoxia is a common tumor phenotype, it is by no means universal and is often heterogeneous within an individual patient. This review considers the biology of hypoxia, its consequences with respect to treatment, methods for measuring oxygenation in tissues, modern techniques for imaging of regional hypoxia, and how information about the oxygenation status of tumors might impact treatment.


Confidential reporting of patient safety events in primary care: results from a multilevel classification of cognitive and system factors.
To classify events of actual or potential harm to primary care patients using a multilevel taxonomy of cognitive and system factors. Observational study of patient safety events obtained via a confidential but not anonymous reporting system. Reports were followed up with interviews where necessary. Events were analysed for their causes and contributing factors using causal trees and were classified using the taxonomy. Five general medical practices in the West Midlands were selected to represent a range of sizes and types of patient population. All practice staff were invited to report patient safety events. Main outcome measures were frequencies of clinical types of events reported, cognitive types of error, types of detection and contributing factors; and relationship between types of error, practice size, patient consequences and detection. 78 reports were relevant to patient safety and analysable. They included 21 (27%) adverse events and 50 (64%) near misses. 16.7% (13/71) had serious patient consequences, including one death. 75.7% (59/78) had the potential for serious patient harm. Most reports referred to administrative errors (25.6%, 20/78). 60% (47/78) of the reports contained sufficient information to characterise cognition: "situation assessment and response selection" was involved in 45% (21/47) of these reports and was often linked to serious potential consequences. The most frequent contributing factor was work organisation, identified in 71 events. This included excessive task demands (47%, 37/71) and fragmentation (28%, 22/71). Even though most reported events were near misses, events with serious patient consequences were also reported. Failures in situation assessment and response selection, a cognitive activity that occurs in both clinical and administrative tasks, was related to serious potential harm.


Novel biphasic separations utilising highly selective molecularly imprinted polymers as biorecognition solvent extraction agents.
Molecularly imprinted polymers (MIPs) represent a class of artificial receptors that promise an environmentally robust alternative to naturally occurring biorecognition elements of biosensing devices and systems. However, in general, the performance of conventional MIPs in aqueous environments is poor. In the study reported here, this limitation has been addressed by the novel application of MIPs as a solvent extraction solid phase in a biphasic solvent system. This paper describes a previously unreported use of MIPs as solvent extraction reagents, their successful application to aqueous sample media and the opportunities for utilisation of this unique system in novel biosensing and separation procedures. This study demonstrates the development of a novel biphasic solvent system utilising MIP in the extracting phase to enhance both efficiency and selectivity of a simple two phase liquid extraction. Monodisperse propranolol imprinted polymer microspheres [p(divinylbenzene-co-methacrylic acid)] were prepared by precipitation polymerisation. Initially, the affinity of the polymers for (R,S)-propranolol was assessed by established techniques whereby the MIP demonstrated greater affinity for the template than did the non-imprinted control polymer (NIP). Importantly, MIP performance was also assessed using the novel dual solvent system. The depletion of (R,S)-propranolol from the aqueous phase into the polymer containing organic phase was determined. When compared to control extractions containing no polymer the presence of MIP in the extracting solvent phase resulted in an increased extraction of (R,S)-propranolol from the aqueous phase. Importantly, this extraction was significantly greater in the presence of MIP when compared to NIP. This unique principle generates opportunities for MIP based extractions and chemical enrichments in industrial applications, offering commercial, ecological and practical advantages to traditional solvent extraction techniques. The technique is readily transferable to analytical microsystems utilising MIP recognition elements generating promising opportunities for MIP based sensing of aqueous sample media.


Utilization of spinal cord stimulation in patients with failed back surgery syndrome.
Retrospective analysis of a population-based insurance claims data set. To evaluate the use of spinal cord stimulation (SCS) and lumbar reoperation for the treatment of failed back surgery syndrome (FBSS), and examine their associated complications and health care costs. FBSS is a major source of chronic neuropathic pain and affects up to 40% of patients who undergo lumbosacral spine surgery for back pain. Thus far, few economic analyses have been performed comparing the various treatments for FBSS, with these studies involving small sample sizes. In addition, the nationwide practices in the use of SCS for FBSS are unknown. The MarketScan data set was used to analyze patients with FBSS who underwent SCS or spinal reoperation between 2000 and 2009. Propensity score methods were used to match patients who underwent SCS with those who underwent lumbar reoperation to examine health care resource utilization. Postoperative complications were analyzed with multivariate logistic regression. Health care use was analyzed using negative binomial and general linear models. The study cohort included 16,455 patients with FBSS, with 395 undergoing SCS implantation (2.4%). Complication rates at 90 days were significantly lower for SCS than spinal reoperation (P < 0.0001). Also in the matched cohort, hospital stay (P < 0.0001) and associated charges (P = 0.016) were lower for patients with SCS. However outpatient, emergency room, and medication charges were similar between the 2 groups. Overall cost totaling $82,586 at 2 years was slightly higher in the lumbar reoperation group than in the SCS group with total cost of $80,669 (P = 0.88). Although previous studies have demonstrated superior efficacy for the treatment of FBSS, SCS remains underused. Despite no significant decreases in overall health care cost with SCS implantation, because it is associated with decreased complications and improved outcomes, this technology warrants closer consideration for the management of chronic pain in patients with FBSS.


Creating a high-performance system for comparative effectiveness research.
Enactment of the federal stimulus and health reform legislation heralds the beginning of a national comparative effectiveness research program. This article suggests how the Department of Health and Human Services (HHS) can, with collaborators, build a high-performing comparative effectiveness research system. New policies and investments should exploit the rapid-learning potential of electronic health records, computerized databases, data sharing, and research networks. A national database for effectiveness research studies should be established by presidential order. HHS will need to support all of these pieces and take the lead in creating a rapid-learning culture for the US health system.


Peritoneal dialysis and kidney transplant. A two-way ticket in an integrated renal replacement therapy model.
Peritoneal dialysis (PD) seems to be a good option to initiate renal replacement therapy (RRT), but patients with graft failure choose PD less frequently than incident patients (de novo). To describe patient movements between PD and kidney transplantation (TX) and risk factors for failure of the PD technique. Multicentre observational study of patients starting PD between 2003 and 2009 with follow-up up until January 2010. Survival analysis based on switching from PD to HD as an event using Kaplan-Meier (KM) and forward, stepwise Cox proportional hazards models. Hazard ratio and 95% confidence intervals (HR [CI]) are shown. MAIN VARIABLE: Switch from PD to HD. Two-group comparison: PD post transplant (post-TX) patients (76) compared to pure incident PD (de novo-PD) patients (830). 906 PD patients from 19 public hospitals with a mean age of 54.8 years (64.9% male); main ESRD aetiology: glomerulonephritis (25.4%), diabetes (16.7%), vascular-ischaemic (10.7%), interstitial (13.6%) and polycystic (11.2%). Comorbidity conditions: Charlson Index 5.1 (SD 2.4); 21.6% diabetes mellitus (DM), 24.0% cardiovascular (CV) events. Mean follow-up period on PD: 1.85 years (95% CI [1.68-2.02 years]). KM estimation for switching to HD due to PD failure was 5.46 years [4.42-6.50 years]. At the end of follow-up, 88 patients had died, 154 had been transferred to HD and 306 had received a graft (annual rate for patients on waiting list: 0.49 TX per year on PD). The best Cox multivariate model for switching from PD to HD includes: post-TX (HR: 1.63 [1.01-2.63]), DM (HR: 1.69 [1.19-2.40]) and age (1.01 [1.00-1.02]) per year. Post-TX patients were younger (43.8 years vs 55.3 years) and with less comorbidity conditions than de novo-PD patients (DM 18.4% vs 21.9%; CV 15.8% vs 24.7%). However post-TX patients had worse clinical evolution with a rapid decline of renal function (∆-3.88 vs -1.8 ml/min per year); a higher admission rate (0.9 vs 0.62 per year) but similar peritonitis rate (0.45 vs 0.53 episodes per year). They also needed to be transferred to HD more frequently (28.9% vs 15.8%; P<.006) and needed more time to TX (4.8 years vs 1.7 years, Kaplan-Meier). Consequently, time spent on PD was higher in the post-TX group (2.8 vs 1.8 year). Observational study with absence of a standard protocol to switch PD-HD. PD seems to be a good first choice technique due to low mortality and high TX ratio in our area. A previous graft failure is associated with a higher rate of PD-failure but time spent on PD is enough to consider this technique as a good option.


Evaluation of the potential effects of ingredients added to cigarettes. Part 2: chemical composition of mainstream smoke.
Cigarette mainstream smoke from blended research cigarettes with and without the addition of ingredients was analyzed for its chemical composition. In total, 333 ingredients commonly used in cigarette manufacturing were assigned to three different groups. Each group of ingredients was introduced at a low and a high level to the test cigarettes. The list of the 51 smoke constituents determined is based on those analytes suggested for analysis in a US Consumer Product Safety Commission proposal for low ignition cigarettes and cigarette smoke constituents identified by the International Agency for Research on Cancer as worthy of concern and characterized as carcinogens. An increase in the yield of total particulate matter (TPM) in the range of 13 to 28% relative to the control cigarette without ingredients was observed for all test cigarettes. This was presumably caused by the higher transfer rates of the added ingredients to the smoke compared to the transfer from the tobacco part of the filler. When the yields of individual constituents were normalized to the TPM yields, a reduction in the majority of the constituents was observed when compared to the control. For one of the ingredient groups this reduction was especially high: for phenols a maximum of 70%, for polycyclic aromatic hydrocarbons 50%, and for N-nitrosamines 45%. An increase in the amount relative to TPM was observed for a few smoke constituents: hydrogen cyanide and cadmium (one ingredient group), formaldehyde (one ingredient group), and resorcinol and lead (two ingredient groups). These results are consistent with the lack of any increased activity in the in vitro and in vivo assays in this same series of studies (Food and Chemical Toxicology 2002, 40, 105-111; Food and Chemical Toxicology 2002, 40, 113-131). An overall assessment of our data suggests that these ingredients, when added to the tobacco, do not add to the toxicity of smoke, even at the elevated levels tested in this series of studies.


Impaired gastric relaxation in patients with achalasia.
Achalasia is considered a primary motility disorder confined to the oesophagus. The lower oesophageal sphincter (LOS) in achalasia is frequently hypertonic and manifests absent or incomplete relaxation in response to deglution. On the other hand, the LOS and the proximal stomach act physiologically as a functional unit whereby relaxation of the LOS during deglution is associated with receptive relaxation of the proximal stomach. Thus, this study investigated the hypothesis that impaired LOS relaxation in patients with achalasia might be associated with impaired relaxation of the proximal stomach. The study consisted of 20 patients with achalasia and 10 healthy controls. Gastric tone variations were quantified using an electronic barostat. Firstly, the study established the basal gastric tone (intragastric volume at the minimal distending pressure+1 mm Hg) and gastric compliance (volume/pressure relation) during isobaric distension (increasing stepwise the intragastric pressure from 0 to 20 mm Hg up to 600 ml). Secondly, the gastric tone response to cold stress (hand immersion into ice water for five minutes) or to control stimuli (water at 37 degrees) was determined. Basal gastric tone mean (SEM) was similar in achalasia and in healthy controls (125 (9) ml v 138 (9) ml, respectively). Compliance was linear and similar in both groups, which also showed similar gastric extension ratios (58 (7) ml/mm Hg v 57 (6) ml/mm Hg). Cold stress induced a gastric relaxatory response that, as a group, was significantly lower in achalasia than in healthy controls (volume: 43 (20) ml v 141 (42) ml; p < 0.05). The responses in each group were not uniform, five of the 20 patients with achalasia showed definite (volume > 100 ml) relaxatory responses whereas four of the 10 healthy controls did not. In conclusion, reflex gastric relaxation is impaired in most patients with achalasia showing that the proximal stomach, and not exclusively the oesophagus, may be effected by the disease.


Structuring feedback and debriefing to achieve mastery learning goals.
Mastery learning is a powerful educational strategy in which learners gain knowledge and skills that are rigorously measured against predetermined mastery standards with different learners needing variable time to reach uniform outcomes. Central to mastery learning are repetitive deliberate practice and robust feedback that promote performance improvement. Traditional health care simulation involves a simulation exercise followed by a facilitated postevent debriefing in which learners discuss what went well and what they should do differently next time, usually without additional opportunities to apply the specific new knowledge. Mastery learning approaches enable learners to "try again" until they master the skill in question. Despite the growing body of health care simulation literature documenting the efficacy of mastery learning models, to date insufficient details have been reported on how to design and implement the feedback and debriefing components of deliberate-practice-based educational interventions. Using simulation-based training for adult and pediatric advanced life support as case studies, this article focuses on how to prepare learners for feedback and debriefing by establishing a supportive yet challenging learning environment; how to implement educational interventions that maximize opportunities for deliberate practice with feedback and reflection during debriefing; describing the role of within-event debriefing or "microdebriefing" (i.e., during a pause in the simulation scenario or during ongoing case management without interruption), as a strategy to promote performance improvement; and highlighting directions for future research in feedback and debriefing for mastery learning.


Poor immune reconstitution is associated with symptomatic BK polyomavirus viruria in allogeneic stem cell transplant recipients.
BK polyomavirus (BKPyV) infections are known indicators of immune suppression in hematopoietic stem cell transplant (HSCT) recipients; they can lead to hemorrhagic cystitis, ureteral stenosis, renal dysfunction, and prolonged hospital stays. In this study, we determined transplant-associated variables and immune parameters that can predict for the risk of BKPyV viruria. We hypothesized that BKPyV infection is a marker of poor immune recovery. We analyzed all engrafted patients undergoing first allogeneic HSCT at MD Anderson Cancer Center in Houston between January 2004 and December 2012. We evaluated their immune parameters and their transplant-associated factors. BKPyV positivity was defined as BKPyV detection in urine by polymerase chain reaction testing. Cox proportional hazards model, as well as competing risk analysis method using subdistribution hazard models with death as competing risk, were applied to assess risk of BKPyV viruria. We identified a total of 2477 patients with a median age of 52 years. BKPyV viruria was manifest in 25% (n=629) of the patients. The median time from transplantation to BKPyV viruria development was 42 days among the patients who had BKPyV viruria. On multivariate analysis, tumor type, acute GVHD, chronic GVHD, myeloablative conditioning regimen, cord blood as the graft source, CD3+ , CD4+ , CD8+ , CD56+ , NK counts, and low platelet count were shown to be significantly associated with BKPyV infection. These finding were further confirmed when models incorporating the competing risk of death yielded similar findings. In this study, we report significant associations between BKPyV reactivation following allogeneic HSCT and suppressed immune variables. In addition, this study provides valuable information on the immune status of HSCT recipients as a predictor of BKPyV infections that may in turn help us formulate plans for more effective prevention and treatment of this infection.


Antitumor effect of gold as revealed by growth suppression of cultured cancer cells.
Gold agents have been widely used for the treatment of rheumatoid arthritis. We studied the growth inhibiting effect of such an agent on malignant cells in vitro. HCT-15, AGS cells derived from a human malignancy, and Meth/A cells from a malignant lymphoma of Balb/C mice were cultured separately with gold agent at concentrations of 2 micrograms/ml. Four days after the cultures had been incubated in a 5% CO2 incubator at 37 degrees C, cell counts were made; and significance of differences was analyzed by Student's t test. Additionally, HCT-15 cells were cultured with gold for two days, and then the cells were analyzed by flow cytometry. The growth of HCT-15, AGS, and Meth/A cells was suppressed by gold. Fifty percent suppression was observed at a concentration between 50 micrograms/ml and 10 micrograms/ml for HCT-15 cells, between 125 micrograms/ml and 50 micrograms/ml for AGS cells, and between 125 micrograms/ml and 50 micrograms/ml for Meth/A cells. Fifty percent suppression of HCT-15 cell growth by cisplatinum was found between 50 micrograms/ml and 10 micrograms/ml. Flow cytometric findings showed a significant rise in the tetraploid peak, a mild rise in the resion between diploid and tetraploid peaks, and an increase in cells with a ploidy greater than four. These data suggest that gold blocks the S phase, G2 to M phase, and M phase as well. To observe the cytotoxicity of gold, each of 10 of 4 week-old Balb/C mice was injected s.c. at a dose of 10 mg/kg or 2 mg/kg every other day for a total of 3 injections, or was administered the gold at 30 mg/kg/day p.o. for 10 days. All mice were still alive after 20 days of observation. Cisplatinum at a dose of 10 mg/kg was also injected s.c. one time into each of 10 mice, and 60% of the animals died within 10 days after the injection.


Genetic characterisation of dough rheological properties in a wheat doubled haploid population: additive genetic effects and epistatic interactions.
Doubled haploid lines (n = 160) from a cross between wheat cultivars 'Cranbrook' (high dough extensibility) and 'Halberd' (low dough extensibility) were grown at three Australian locations. The parents differ at all high- and low-molecular-weight glutenin loci. Dough rheological parameters were measured using small-scale testing procedures, and quantitative trait locus (QTL) mapping procedures were carried out using an existing well-saturated genetic linkage map for this cross. Genetic parameters were estimated using three software packages: QTLCartographer, Epistat and Genstat. Results indicated that environmental factors are a major determinant of dough extensibility across the three trial sites, whereas genotypic factors are the major determinants of dough strength. Composite interval mapping analysis across the 21 linkage groups revealed that as expected, the main additive QTLs for dough rheological properties are located at the high- and low-molecular-weight glutenin loci. A new QTL on chromosome 5A for M-extensibility (a mixograph-estimated measure of extensibility) was detected. Analysis of epistatic interactions revealed that there were significant conditional epistatic interactions related with the additive effects of glutenin loci on dough rheological properties. Therefore, the additive genetic effects of glutenins on dough rheological properties are conditional upon the genetic background of the wheat line. The molecular basis of the interactions with the glutenin loci may be via proteins that modify or alter the gluten protein matrix or variations in the expression level of the glutenin genes. Reverse-phase high performance liquid chromatography analysis of the molar number of individual glutenin subunits across the population showed that certain conditional epistases resulted in increased expression of the affected glutenin. The epistatic interactions detected in this study provide a possible explanation of the variable genetic effects of some glutenins on quality attributes in different genetic backgrounds and provide essential information for the accurate prediction of glutenin related variance in marker-assisted wheat breeding.


The seroepidemiology of rubella in western Europe.
Most of the countries in western Europe have now implemented mass infant rubella immunization programmes, instead of or in addition to selective vaccination in order to achieve the elimination of congenital rubella syndrome. The European countries Denmark, England and Wales, Finland, France, Germany, Italy and the Netherlands undertook large, national serological surveys collecting several thousand serum specimens during 1994-8. Antibodies against rubella virus were detected by a variety of enzyme immuno-assays. Comparability of the assay results was achieved by a standardized methodology. The age- and sex-stratified serological results were related to the schedules, coverage of rubella vaccination and the incidence in these countries. The results show widely differing levels of immunity to rubella both in the general population and in the specific age groups of males and females. A low rate (< 5%) of susceptibles in childhood and adolescents of both sexes was obtained only in Finland and the Netherlands. Countries such as Italy with only moderate coverage for the infant immunization programme currently have both high susceptibility levels in the general population and in the at-risk population. The likelihood is of continued epidemics of rubella with cases of congenital rubella syndrome. The continued implementation of selective vaccination will help to offset the impact of this ongoing transmission and to protect women on reaching childbearing age.


HIV/AIDS affects blood and blood product use at Groote Schuur Hospital, Cape Town.
Use of blood and blood products in the medical wards at Groote Schuur Hospital, Cape Town, has increased substantially and significantly increased expenditure. It was suspected that the increased burden of HIV/AIDS could be a contributing factor. Doctors voluntarily completed a structured questionnaire when blood or blood products were utilised over a 3-month period in 2009. Statistical analysis was performed using Microsoft Excel, SPSS and STATISTICA. Of 67 patients analysed, 46 (68.6%) were female, mean age 36.7 (standard deviation (SD) 8.7) years; 21 (31.3%) were male, mean age 39.3 (SD 13.5) years; and 41 (61.2%) were HIV positive, of whom 17 (41.5%) were on antiretroviral therapy (ART). HIV-infected patients were on average 10 years younger than HIV-uninfected patients (p=0.012). Anaemia was the cytopenia necessitating transfusion in 68.7% of cases, but its causes differed between HIV-infected and uninfected patients. The median CD4 count was 203 cells/ml (range 24 - 540) for HIV-infected patients on ART and 74 cells/ml (range 2 - 276) for those not on ART (p=0.012). The mean numbers of packed red cell and fresh-frozen plasma units transfused in the HIV-infected not on ART, HIV-infected on ART and HIV-uninfected groups were 3.3, 2.0 and 1.5 (p=0.013) and 13.5, 2.7 and 1.0 (p.<0.001), respectively. ART in HIV-positive patients markedly decreased transfusion requirements (p<0.001). There was one minor transfusion reaction. HIV/AIDS is a significant factor contributing to the increased use of blood and blood products in the medical wards at Groote Schuur Hospital. Being on ART appeared to reduce the requirement for blood and blood products.


Large-area atomically thin MoS2 nanosheets prepared using electrochemical exfoliation.
Molybdenum disulfide (MoS2) is an extremely intriguing material because of its unique electrical and optical properties. The preparation of large-area and high-quality MoS2 nanosheets is an important step in a wide range of applications. This study demonstrates that monolayer and few-layer MoS2 nanosheets can be obtained from electrochemical exfoliation of bulk MoS2 crystals. The lateral size of the exfoliated MoS2 nanosheets is in the 5-50 μm range, which is much larger than that of chemically or liquid-phase exfoliated MoS2 nanosheets. The MoS2 nanosheets undergo low levels of oxidation during electrochemical exfoliation. In addition, microscopic and spectroscopic characterizations indicate that the exfoliated MoS2 nanosheets are of high quality and have an intrinsic structure. A back-gate field-effect transistor was fabricated using an exfoliated monolayer MoS2 nanosheet. The on/off current ratio is over 10(6), and the field-effect mobility is approximately 1.2 cm(2) V(-1) s(-1); these values are comparable to the results for micromechanically exfoliated MoS2 nanosheets. The electrochemical exfoliation method is simple and scalable, and it can be applied to exfoliate other transition metal dichalcogenides.


Informational gene phylogenies do not support a fourth domain of life for nucleocytoplasmic large DNA viruses.
Mimivirus is a nucleocytoplasmic large DNA virus (NCLDV) with a genome size (1.2 Mb) and coding capacity ( 1000 genes) comparable to that of some cellular organisms. Unlike other viruses, Mimivirus and its NCLDV relatives encode homologs of broadly conserved informational genes found in Bacteria, Archaea, and Eukaryotes, raising the possibility that they could be placed on the tree of life. A recent phylogenetic analysis of these genes showed the NCLDVs emerging as a monophyletic group branching between Eukaryotes and Archaea. These trees were interpreted as evidence for an independent "fourth domain" of life that may have contributed DNA processing genes to the ancestral eukaryote. However, the analysis of ancient evolutionary events is challenging, and tree reconstruction is susceptible to bias resulting from non-phylogenetic signals in the data. These include compositional heterogeneity and homoplasy, which can lead to the spurious grouping of compositionally-similar or fast-evolving sequences. Here, we show that these informational gene alignments contain both significant compositional heterogeneity and homoplasy, which were not adequately modelled in the original analysis. When we use more realistic evolutionary models that better fit the data, the resulting trees are unable to reject a simple null hypothesis in which these informational genes, like many other NCLDV genes, were acquired by horizontal transfer from eukaryotic hosts. Our results suggest that a fourth domain is not required to explain the available sequence data.


A rare case of mucormycosis in the scalp.
Mucormycosis is a rapidly progressive and frequently lethal form of fungal infection. Primary cutaneous form is rare. We presented a case of cutaneous mucormycosis in the scalp which led to visual loss in a 54-year-old diabetic woman. A 54-year-old woman patient was admitted with the complaining of the wound with purulent discharge in the scalp. An ulcerated discharging lesion with necrotic hemorrhagic crusts in the left parietal region of the scalp and wheals with fluctuating from this lesion to the left periorbital area were observed. Biochemical investigations revealed elevated blood sugar level and urine ketone bodies were present. The patient had vision loss in the left eye. In the smears thick-walled non-septate hyphae were detected. Rhizopus spp. was isolated from culture. Antidiabetic therapy and liposomal amphotericin B was initiated. The scalp lesion improved. Early diagnosis is life saving in mucormycosis.


Cytomorphology of primary small-cell (Merkel-cell) carcinoma of the skin in fine needle aspirates.
The histologic appearance of primary small-cell carcinoma of the skin (the so-called Merkel-cell tumor) is similar to other small-cell tumors that may metastasize to the dermis. Significance has been placed on the electron microscopic appearance of this tumor since the ultrastructural features of this neoplasm are helpful in distinguishing it from most of the other neoplasms considered in the differential diagnosis. To determine whether any additional morphologic criteria might exist to distinguish this neoplasm, the fine needle aspirate appearance of a primary small-cell carcinoma of the skin was studied and compared to that of similar preparations of other small-cell tumors that could potentially involve the dermis. Cells of this unusual tumor were round and showed neither cohesiveness nor nuclear molding. Mitoses were numerous. The chromatin pattern was bland. The cytologic features of this tumor can aid in the distinction of primary small-cell carcinoma of the skin from other metastatic small-cell neoplastic lesions in the dermis of adults.


Bradysomnia in Parkinson's disease.
Polysomnography studies in Parkinson's disease (PD) patients show altered sleep microstructure with decreased level of arousability, indicating impaired sleep-wake dynamics in PD. The aim of this study was to investigate dynamical aspects of sleep EEG in PD as compared to healthy controls. In this retrospective, controlled study, we applied a previously established mathematical model of sleep EEG analysis (state space model) to PD patients and age- and gender-matched healthy volunteers (N=64). Dynamical aspects of sleep were quantified by measuring the spectral variability of the sleep EEG (by means of state space velocity). State space analysis revealed preserved global sleep-wake architecture in PD patients, but the velocity of sleep stage transitions was significantly reduced as compared to healthy controls. Correlation analysis revealed a strong association of state space velocity with arousal scores and daily dopamine agonist intake. Quantitative analysis of spectral sleep EEG variability (state space velocity) revealed reduced sleep-wake dynamics in PD patients as compared to control subjects. We propose state space velocity as an objective and quantitative measure for altered sleep microstructure and as a potential biomarker of sleep alterations in PD, not accessible by conventional sleep analysis.


LPV control of glucose for Diabetes type I.
This paper considers the problem of automatically controlling the glucose level in a Diabetes type I patient. Three issues have been considered: model uncertainty, time-varying/nonlinear phenomena and controller implementation. To that end, the dynamical model of the insulin/glucose relation is framed as a Linear Parameter Varying system and a controller is designed based on it. In addition, this framework allows not only a better performance than other classical methods, but also provides stability and performance guarantees. Design computations are based on convex Linear Matrix Inequality (LMI) optimization. Implementation is based on a low order controller whose dynamics adapts according to the glucose levels measured in real-time.


Experimental study of the inhibition effect of CXCL12/CXCR4 in malignant pleural mesothelioma.
Previous studies have demonstrated that CXCL12/CXCR4 axis is closely related to tumors such as malignant pleural mesothelioma (MPM). This research was conducted in order to detect whether CXCL12/CXCR4 inhibitors could restrain MPM and have a synergistic effect with chemotherapy, also to investigate the relationship of CXCL12/CXCR4 with other gene expressions in MPM. Forty mice were injected MPM cells and randomly divided into four groups: the PBS (control group), AMD3100 (CXCR4-CXCL12 antagonist), pemetrexed and AMD3100 plus pemetrexed. The mice were treated respectively for duration of 3 weeks. The size, bioluminescence and weight of tumors were measured. The differences between gene expressions in each group were analyzed. The tumor weights of each treatment group were lower than that of the control group (p<0.05). The bioluminescence of the tumor of the AMD3100 treatment group and the AMD3100 plus pemetrexed treatment group were lower than that of the control group (p<0.05), and AMD3100 was shown to have synergistic effects with pemetrexed (p<0.05). Among the 2.5 billion genes, several hundreds of genes expressed differently between groups. Results show that AMD3100 and pemetrexed can inhibit the growth of MPM in vivo, also that there is a better result if both are used together. Our findings suggest that CXCL12/CXCR4 axis affects a certain amount of gene expression in MPM.


Inhibitory effects of taraxasterol and aqueous extract of Taraxacum officinale on calcium oxalate crystallization: in vitro study.
We investigated and compared the effects of taraxasterol, aqueous extract of T. officinale (AET) aerial part, and potassium citrate (PC) on calcium oxalate (CaOx) crystallization in vitro. CaOx crystallization was induced by adding sodium oxalate to synthetic urine. Taraxasterol (2.5, 5, 7.5 and 12.5 μg/mL), extract (1, 2, 4 and 8 mg/mL), and PC (100, 150, 200 and 350 mg/mL) were subjected to anti-crystallization activities. The absorbance and %inhibition of nucleation of CaOx crystals were evaluated by spectrophotometer at 2, 4, 6, 8, 10, 20, 30, 40, 50 and 60 min and the number and morphology of crystals were studied by light microscopy after 60 min. Presence of taraxasterol, extract and PC decreased absorbance in experimental samples compared to control, significantly. The nucleation of crystals is inhibited by taraxasterol, extract, and PC (26-64, 55-63 and 60-70%, respectively). The number of CaOx crystals were decreased in presence of taraxasterol (p < .01), extract (p < .001), and PC (p < .001) in a dose-dependent manner. Presence of taraxasterol, extract, and PC decreased the number of CaC2O4 monohydrate, while increased CaC2O4 dihydrate crystals, significantly. Also, the diameter of CaC2O4 dihydrate crystals was decreased in presence of taraxasterol, extract and PC, significantly. This research indicated that taraxasterol and extract have anti-crystallization activities and effectiveness of the extract is more potent than taraxasterol. It could be because of another constituent in the extract with the synergistic effect.


Layer speciation and electronic structure investigation of freestanding hexagonal boron nitride nanosheets.
Chemical imaging, thickness mapping, layer speciation and polarization dependence have been performed on single and multilayered (up to three layers and trilayered nanosheets overlapping to form 6 and 9 layers) hexagonal boron nitride (hBN) nanosheets by scanning transmission X-ray microscopy. Spatially-resolved XANES directly from freestanding regions of different layers has been extracted and compared with sample normal and 30° tilted configurations. Notably a double feature σ* excitonic state and a stable high energy σ* state were observed at the boron site in addition to the intense π* excitonic state. The boron projected σ* DOS, especially the first σ* exciton, is sensitive to surface modification, particularly in the single layered hBN nanosheet which shows more significant detectable contaminants and defects such as tri-coordinated boron/nitrogen oxide. The nitrogen site has shown very weak or no excitonic character. The distinct excitonic effect on boron and nitrogen was interpreted to the partly ionic state of hBN. Bulk XANES of hBN nanosheets was also measured to confirm the spectro-microscopic STXM result. Finally, the unoccupied electronic structures of hBN and graphene were compared.


Total dissociative electron attachment cross sections of selected amino acids.
Total dissociative electron attachment cross sections are presented for the amino acids, glycine, alanine, proline, phenylalanine, and tryptophan, at energies below the first ionization energy. Cross section magnitudes were determined by observation of positive ion production and normalization to ionization cross sections calculated using the binary-encounter-Bethe method. The prominent 1.2 eV feature in the cross sections of the amino acids and the closely related HCOOH molecule is widely attributed to the attachment into the -COOH pi* orbital. The authors discuss evidence that direct attachment to the lowest sigma* orbital may instead be responsible. A close correlation between the energies of the core-excited anion states of glycine, alanine, and proline and the ionization energies of the neutral molecules is found. A prominent feature in the total dissociative electron attachment cross section of these compounds is absent in previous studies using mass analysis, suggesting that the missing fragment is energetic H-.