Nanosheet Supported Single-Metal Atom Bifunctional Catalyst for Overall Water Splitting.
Nanosheet supported single-atom catalysts (SACs) can make full use of metal atoms and yet entail high selectivity and activity, and bifunctional catalysts can enable higher performance while lowering the cost than two separate unifunctional catalysts. Supported single-atom bifunctional catalysts are therefore of great economic interest and scientific importance. Here, on the basis of first-principles computations, we report a design of the first single-atom bifunctional eletrocatalyst, namely, isolated nickel atom supported on β12 boron monolayer (Ni1/β12-BM), to achieve overall water splitting. This nanosheet supported SAC exhibits remarkable electrocatalytic performance with the computed overpotential for oxygen/hydrogen evolution reaction being just 0.40/0.06 V. The ab initio molecular dynamics simulation shows that the SAC can survive up to 800 K elevated temperature, while enacting a high energy barrier of 1.68 eV to prevent isolated Ni atoms from clustering. A viable experimental route for the synthesis of Ni1/β12-BM SAC is demonstrated from computer simulation. The desired nanosheet supported single-atom bifunctional catalysts not only show great potential for achieving overall water splitting but also offer cost-effective opportunities for advancing clean energy technology.


Nifedipine retard was as effective as angiotensin converting enzyme inhibitors in preventing cardiac events in high-risk hypertensive patients with diabetes and coronary artery disease: the Japan Multicenter Investigation for Cardiovascular Diseases-B (JMIC-B) subgroup analysis.
We stratified findings from the Japan Multicenter Investigation for Cardiovascular Diseases-B according to whether or not the patients had diabetes and compared the incidence of cardiac events occurring over a 3-year period between treatment with nifedipine retard and angiotensin converting enzyme (ACE) inhibitor. The primary endpoint was the overall incidence of cardiac events (cardiac death or sudden death, myocardial infarction, hospitalization for angina pectoris or heart failure, serious arrhythmia, and coronary interventions), and the secondary endpoints were a composite of other events (cerebrovascular accidents, worsening of renal dysfunction, non-cardiovascular events, and total mortality). The results showed no significant difference in the incidence of the primary endpoint between the nifedipine group (n=199) and the ACE inhibitor group (n=173) in diabetic patients: 15.08% vs. 15.03%, relative risk 1.06, p=0.838. Also in nondiabetic patients, no significant difference was observed between the former (n=629) and the latter (n=649): 13.67% vs. 12.33%, relative risk 1.04, p=0.792. Similar results were obtained for the incidence of the secondary endpoints: in diabetic patients, 5.03% vs. 5.20%, relative risk 0.89, p=0.799; in nondiabetic patients, 2.70% vs. 2.47%, relative risk 1.07, p=0.842. Achieved blood pressure levels were 138/76 and 136/77 mmHg in the nifedipine group and 140/78 and 138/79 mmHg in the ACE inhibitor group in diabetic and nondiabetic patients, respectively. This study showed that nifedipine retard was as effective as ACE inhibitors in reducing the incidence of cardiac events in extremely high-risk hypertensive patients with complications of diabetes and coronary artery disease.


Quantification of retinal pigment epithelial phenotypic variation using laser scanning cytometry.
Quantifying phenotypic variation at the level of protein expression (variegation) within populations of retinal pigment epithelium (RPE) cells may be important in the study of pathologies associated with this variation. The lack of quantitative methods for examining single cells, however, and the variable presence of pigment and/or lipofuscin complicate this experimental goal. We have applied the technique of laser scanning cytometry (LSC) to paraffin sections of mouse and human eyes to evaluate the utility of LSC for these measurements. Mouse eyes were perfusion fixed in 4% paraformaldehyde and embedded in paraffin. Postmortem human eyes were fixed and dissected to obtain a 9-mm punch, which was then embedded in paraffin. A laser scanning cytometer equipped with violet, argon, and helium-neon lasers and the detectors for blue, green, and long red were used to record the fluorescence of each individual cell at all three wavelengths. Raw data were recorded and processed using the WinCyte software. Individual nuclei were identified by the fluorescence of the 4',6-diamidino-2-phenylindole (DAPI) nuclear counterstain. Next, RPE cells were uniquely identified in the green channel using an anti-retinal pigment epithelium-specific protein 65 kDa (anti-RPE65) monoclonal antibody with an Alexa Fluor 488-labeled secondary antibody. Mn-superoxide dismutase (MnSOD) was quantified in the long-red channel using an anti-MnSOD antibody and an Alexa Fluor 647-labeled secondary antibody. MnSOD(+) and RPE65(+) cells exhibited peaks in the plot of fluorescence intensity versus cell number, which could be characterized by the mean fluorescence intensity (MFI), the coefficient of variation (CV), and the percentage of total RPE cells that were also labeled for MnSOD. RPE cells can be uniquely identified in human and mouse paraffin sections by immunolabeling with anti-RPE65 antibody. A second antigen, such as MnSOD, can then be probed only within this set of RPE. Results are plotted primarily with the population frequency diagram, which can be subdivided into multiple regions. The data collected for each region include the MFI, the CV, and the number of cells that are immunolabeled in that region. Background interference from pigment or autofluorescent material can be successfully overcome by elevating the concentrations of fluorescent secondary antibodies. In the human and mouse eyes, age-related changes in MFI, CV, and percent RPE cells immunolabeled for MnSOD were observed. The extent of the variability of gene expression in RPE cells at the protein level can be quantified by LSC. Relative changes in the MFI, the CV, and/or percentage of RPE cells double labeled for a second antigen quantify the changes observed. The analysis of these data also suggest whether the effects observed are related to local changes in transcription (alterations of CV) or major changes of protein expression (MFI), which are likely to be due to changes in the chromatin structure. The changes of these variables with age suggest that the observed age-related variegation is primarily due to changes in the chromatin structure in individual cells.


Third trimester maternal vitamin D and early childhood socioemotional development.
Whether maternal vitamin D affects offspring socioemotional development in early childhood has been underexplored. This study examined associations between maternal vitamin D during in the 3rd trimester and offspring socioemotional development between 30 and 59 months. Data from 87 maternal-offspring pairs enrolled in the National Children's Study were used. Total plasma maternal vitamin D (25-hydroxyergocalciferol + 25-hydroxycholecalciferol) was measured between 28 and 35 gestational weeks and categorised as quartiles (Q). Multivariable regression models, adjusting for maternal race/ethnicity, education, and prepregnancy body mass index (BMI [kg/m2 ]), were used to estimate the association between vitamin D and offspring scores on the Brief Infant-Toddler Social and Emotional Assessment (BITSEA). The mean (standard deviation) vitamin D concentration was 86.5 (27.8) nmol/L. The median (range) BITSEA problem score was 6.0 (0.0-30.0), and competence score was 19.0 (7.0-22.0). Maternal vitamin D was inversely related to offspring problem scores. Compared to offspring of women with 25(OH)D in Q1, offspring problem scores were -4.80 (95% confidence interval [CI] -8.29, -1.33) units lower for Q2 vs Q1, -5.64 (95% CI -9.60, -1.68) units lower for Q3 vs Q1, and -4.70 (95% CI -8.59, -0.82) units lower for Q4 vs Q1. Vitamin D was not associated with offspring competence score. Higher maternal vitamin D was associated with lower offspring behaviour problems and not associated with socioemotional competence. These data indicate the association of maternal vitamin D and offspring development may be dependent on the specific developmental component being investigated.


Medical cannibus as an alternative for opioids for chronic pain: A case report.
Opioid medication-related deaths have increased to epidemic proportions in the last decade. This report describes a case of 43-year-old female with a traumatic brain injury who developed chronic pain and opioid dependence. The patient expressed concerns and wanted weaning off opioids. Recent legalization of medical marijuana in Pennsylvania allows us to try it as an alternative to opioids for chronic pain. Medical cannibus has risks associated with administration but is safer than opioids. Our patient was successfully weaned off her opioid medications with the help of medical cannibus and pain remained well controlled. More studies need to be done on using medical cannibus as an alternative to opioids.


An integrated approach to bovine oocyte quality: from phenotype to genes.
In cattle, early embryonic failure plays a major role in the limitation of reproductive performance and is influenced by genetic effects. Suboptimal oocyte quality, including an inadequate store of maternal factors, is suspected to contribute to this phenomenon. In the present study, 13 Montbeliarde cows were phenotyped on oocyte quality, based on their ability to produce viable embryos after in vitro maturation, fertilisation and culture for 7 days. This discriminated two groups of animals, exhibiting developmental rates below 18.8% or above 40.9% (relative to cleaved embryos). Using microarrays, transcriptomic profiles were compared between oocytes collected in vivo from these two groups of animals. The difference in oocyte development potential was associated with changes in transcripts from 60 genes in immature oocytes and 135 genes in mature oocytes (following Bonferroni 5% correction). Of these, 16 and 32 genes were located in previously identified fertility quantitative trait loci. A subset of differential genes was investigated on distinct samples by reverse transcription-quantitative polymerase chain reaction. For SLC25A16, PPP1R14C, ROBO1, AMDHD1 and MEAF6 transcripts, differential expression was confirmed between high and low oocyte potential animals. Further sequencing and searches for polymorphisms will pave the way for implementing their use in genomic selection.


Extracellular matrix adhesion-promoting activities of a dermatan sulfate proteoglycan-associated protein (22K) from bovine fetal skin.
A 22 x 10(3) Mr protein (abbreviated 22K) that copurifies with dermatan sulfate proteoglycans (DS-PGs) following the biochemical fractionation of bovine fetal skin has been evaluated for adhesion-promoting activity in vitro using Balb/c 3T3 cells, as well as bovine and human dermal fibroblasts. Substrata coated with 22K protein promote attachment of a subset of 3T3 and dermal fibroblasts that respond to plasma fibronectin (pFN) substrata. Cells on 22K protein display partial cytoplasmic spreading, comparable to that of cells adhering to cell-binding fragments of pFN. Adhesion activity of 22K is not due to contamination with known adhesive proteins of dermal matrices and is not dermal cell type-specific, since two classes of neuronal cells also respond effectively to 22K substrata. DS-PGs from cartilage or skin completely inhibit 22K adhesion activity when the PGs are adsorbed to 22K substrata under conditions prohibiting PGs from binding to substrata directly. Cartilage chondroitin/keratan sulfate proteoglycan at much higher concentrations is only partially inhibitory. Inhibition by DS-PGs is mediated by DS chains binding to 22K. Properties of the cell surface 'receptor' for 22K protein were tested by several approaches. It is not cell surface DS-PG, since: (1) cells unable to produce this proteoglycan class also responded; (2) cells treated with chondroitinase ABC responded equally well; and (3) substrata of proteoglycan-binding platelet factor-4 generated responses from cells that were quantitatively and qualitatively different. A synthetic peptide in the medium containing the Arg-Gly-Asp-Ser (RGDS) sequence completely inhibited responses to 22K substrata. This observation, coupled with sequencing data of 22K protein revealing an Arg-Gly-Ala-Thr sequence at residues 151-154, suggest that 22K protein mediates adhesion by cell surface integrin binding. Therefore, this newly discovered matrix protein from skin may serve as a communication link between the dermal fibroblast cell surface and its extracellular matrix environment.


Inflammatory responses in brain ischemia.
Brain infarction causes tissue death by ischemia due to occlusion of the cerebral vessels and recent work has shown that post stroke inflammation contributes significantly to the development of ischemic pathology. Because secondary damage by brain inflammation may have a longer therapeutic time window compared to the rescue of primary damage following arterial occlusion, controlling inflammation would be an obvious therapeutic target. A substantial amount of experimentall progress in this area has been made in recent years. However, it is difficult to elucidate the precise mechanisms of the inflammatory responses following ischemic stroke because inflammation is a complex series of interactions between inflammatory cells and molecules, all of which could be either detrimental or beneficial. We review recent advances in neuroinflammation and the modulation of inflammatory signaling pathways in brain ischemia. Potential targets for treatment of ischemic stroke will also be covered. The roles of the immune system and brain damage versus repair will help to clarify how immune modulation may treat stroke.


Review: Deciphering animal robustness. A synthesis to facilitate its use in livestock breeding and management.
As the environments in which livestock are reared become more variable, animal robustness becomes an increasingly valuable attribute. Consequently, there is increasing focus on managing and breeding for it. However, robustness is a difficult phenotype to properly characterise because it is a complex trait composed of multiple components, including dynamic elements such as the rates of response to, and recovery from, environmental perturbations. In this review, the following definition of robustness is used: the ability, in the face of environmental constraints, to carry on doing the various things that the animal needs to do to favour its future ability to reproduce. The different elements of this definition are discussed to provide a clearer understanding of the components of robustness. The implications for quantifying robustness are that there is no single measure of robustness but rather that it is the combination of multiple and interacting component mechanisms whose relative value is context dependent. This context encompasses both the prevailing environment and the prevailing selection pressure. One key issue for measuring robustness is to be clear on the use to which the robustness measurements will employed. If the purpose is to identify biomarkers that may be useful for molecular phenotyping or genotyping, the measurements should focus on the physiological mechanisms underlying robustness. However, if the purpose of measuring robustness is to quantify the extent to which animals can adapt to limiting conditions then the measurements should focus on the life functions, the trade-offs between them and the animal's capacity to increase resource acquisition. The time-related aspect of robustness also has important implications. Single time-point measurements are of limited value because they do not permit measurement of responses to (and recovery from) environmental perturbations. The exception being single measurements of the accumulated consequence of a good (or bad) adaptive capacity, such as productive longevity and lifetime efficiency. In contrast, repeated measurements over time have a high potential for quantification of the animal's ability to cope with environmental challenges. Thus, we should be able to quantify differences in adaptive capacity from the data that are increasingly becoming available with the deployment of automated monitoring technology on farm. The challenge for future management and breeding will be how to combine various proxy measures to obtain reliable estimates of robustness components in large populations. A key aspect for achieving this is to define phenotypes from consideration of their biological properties and not just from available measures.


Comparative analysis of Pareto surfaces in multi-criteria IMRT planning.
In the multi-criteria optimization approach to IMRT planning, a given dose distribution is evaluated by a number of convex objective functions that measure tumor coverage and sparing of the different organs at risk. Within this context optimizing the intensity profiles for any fixed set of beams yields a convex Pareto set in the objective space. However, if the number of beam directions and irradiation angles are included as free parameters in the formulation of the optimization problem, the resulting Pareto set becomes more intricate. In this work, a method is presented that allows for the comparison of two convex Pareto sets emerging from two distinct beam configuration choices. For the two competing beam settings, the non-dominated and the dominated points of the corresponding Pareto sets are identified and the distance between the two sets in the objective space is calculated and subsequently plotted. The obtained information enables the planner to decide if, for a given compromise, the current beam setup is optimal. He may then re-adjust his choice accordingly during navigation. The method is applied to an artificial case and two clinical head neck cases. In all cases no configuration is dominating its competitor over the whole Pareto set. For example, in one of the head neck cases a seven-beam configuration turns out to be superior to a nine-beam configuration if the highest priority is the sparing of the spinal cord. The presented method of comparing Pareto sets is not restricted to comparing different beam angle configurations, but will allow for more comprehensive comparisons of competing treatment techniques (e.g., photons versus protons) than with the classical method of comparing single treatment plans.


Synergistic effects of ultrasound and sodium hypochlorite (NaOCl) on reducing Listeria monocytogenes ATCC19118 in broth, stainless steel, and iceberg lettuce.
This study was performed in order to determine whether a combined treatment of ultrasound and sodium hypochlorite (NaOCl) is more effective than individual treatment on reducing Listeria monocytogenes ATCC19118 on stainless steel and iceberg lettuce. The bactericidal effect of ultrasound and NaOCl was investigated in tryptic soy broth (TSB), on stainless steel and iceberg lettuce. Various concentrations of NaOCl (50, 100, 150, and 200 ppm) were tested along with various ultrasound treatment times (5, 20, 40, 60, 80, and 100 min). The combined treatment of ultrasound and NaOCl resulted in greater bacterial reductions than either treatment alone, without causing any significant changes in lettuce texture. The synergistic values of combined ultrasound and NaOCl treatments in TSB, on stainless steel, and on iceberg lettuce were 0.01-0.99 log10 colony-forming units (CFU)/mL, 0.01-0.62 log 10 CFU/g, and 0.12-1.66 log10 CFU/g, respectively. These results suggest that the combination of ultrasound and NaOCl was more effective than each treatment against Listeria monocytogenes, and that this combination can effectively sanitize fresh products such as iceberg lettuce.


Structure determination of symmetric homo-oligomers by a complete search of symmetry configuration space, using NMR restraints and van der Waals packing.
Structural studies of symmetric homo-oligomers provide mechanistic insights into their roles in essential biological processes, including cell signaling and cellular regulation. This paper presents a novel algorithm for homo-oligomeric structure determination, given the subunit structure, that is both complete, in that it evaluates all possible conformations, and data-driven, in that it evaluates conformations separately for consistency with experimental data and for quality of packing. Completeness ensures that the algorithm does not miss the native conformation, and being data-driven enables it to assess the structural precision possible from data alone. Our algorithm performs a branch-and-bound search in the symmetry configuration space, the space of symmetry axis parameters (positions and orientations) defining all possible C(n) homo-oligomeric complexes for a given subunit structure. It eliminates those symmetry axes inconsistent with intersubunit nuclear Overhauser effect (NOE) distance restraints and then identifies conformations representing any consistent, well-packed structure to within a user-defined similarity level. For the human phospholamban pentamer in dodecylphosphocholine micelles, using the structure of one subunit determined from a subset of the experimental NMR data, our algorithm identifies a diverse set of complex structures consistent with the nine intersubunit NOE restraints. The distribution of determined structures provides an objective characterization of structural uncertainty: backbone RMSD to the previously determined structure ranges from 1.07 to 8.85 A, and variance in backbone atomic coordinates is an average of 12.32 A(2). Incorporating vdW packing reduces structural diversity to a maximum backbone RMSD of 6.24 A and an average backbone variance of 6.80 A(2). By comparing data consistency and packing quality under different assumptions of oligomeric number, our algorithm identifies the pentamer as the most likely oligomeric state of phospholamban, demonstrating that it is possible to determine the oligomeric number directly from NMR data. Additional tests on a number of homo-oligomers, from dimer to heptamer, similarly demonstrate the power of our method to provide unbiased determination and evaluation of homo-oligomeric complex structures.


Do Adolescents Use Substances to Relieve Uncomfortable Sensations? A Preliminary Examination of Negative Reinforcement among Adolescent Cannabis and Alcohol Users.
Alcohol and cannabis use are highly prevalent among adolescents and associated with negative consequences. Understanding motivations behind substance use in youth is important for informing prevention and intervention efforts. The present study aims to examine negative reinforcement principles of substance use among adolescent cannabis and alcohol users by pairing a cue reactivity paradigm with an aversive interoceptive stimulus. Adolescents (ages 15-17), classified as controls (CTL; n = 18), cannabis and/or alcohol experimenters (CAN+ALC-EXP; n = 16), or individuals meeting clinical criteria for cannabis and/or alcohol use disorder (CAN+ALC-SUD; n = 13) underwent functional magnetic resonance imaging during which they experienced an aversive interoceptive probe delivered via breathing load while simultaneously performing a cue reactivity paradigm. Participants also provided self-report ratings of how their substance use is positively or negatively reinforced. While experiencing the breathing load, CAN+ALC-SUD exhibited greater (p < 0.05) deactivation in the right amygdala, the left inferior frontal gyrus, and the left parahippocampal gyrus than CAN+ALC-EXP and CTL, who did not differ. Across all substance users, activation during the breathing load within the left parahippocampal gyrus negatively correlated with cannabis and alcohol lifetime use episodes and the left inferior frontal gyrus activity negatively correlated with lifetime alcohol use episodes. CAN+ALC-SUD reported experiencing more positive and negative reinforcement of using their substance of choice than CAN+ALC-EXP; both user groups reported higher levels of positive than negative reinforcement. Adolescents with a cannabis/alcohol use disorder demonstrate an altered response to interoceptive perturbations. However, adolescent cannabis/alcohol use does not appear to be driven by negative reinforcement, as viewing substance images did not dampen this response. Based on self-report data, the experience of positive reinforcement may be stronger for adolescents. Future studies should examine whether positive reinforcement contributes to adolescent substance use.


Advancing the Science of Deprescribing: A Novel Comprehensive Conceptual Framework.
Polypharmacy is common in older adults and associated with inappropriate medication use, adverse drug events, medication nonadherence, higher costs, and increased mortality compared with those without polypharmacy. Deprescribing, the clinically supervised process of stopping or reducing the dose of medications when they cause harm or no longer provide benefit, may improve outcomes. Although potentially beneficial, clinicians struggle to overcome structural, organizational, technological, and cognitive barriers to deprescribing, limiting its use in clinical practice. Deprescribing science would benefit from a unifying conceptual framework to prioritize research. Current deprescribing conceptual frameworks have made important contributions to the field but often with a focus on specific medication classes or aspects of deprescribing. To further this relatively nascent field, we developed a broader deprescribing conceptual framework that builds on prior frameworks and includes patient, prescriber, and system influences; the process of deprescribing; outcomes; and dissemination. Patient factors include patients' biology, experience, values, and preferences. Prescriber factors include rational (eg, based on explicit knowledge) and nonrational (eg, behavioral tendencies, biases, and heuristics) decision making. System factors include resources, incentives, goals, and culture that contribute to deprescribing. The framework separates the deprescribing decision from the deprescribing process. The framework captures the results of deprescribing by examining changes in clinical structures, performance processes, patient experience, health outcomes, and cost. Through testing and refinement, this novel, more comprehensive conceptual framework has the potential to advance deprescribing research by organizing the existing evidence, identifying evidence gaps, and categorizing deprescribing interventions and the settings in which they are applied. J Am Geriatr Soc 67:2018-2022, 2019.


Matrix in River Water, Sediments, and Biofilms Mitigates Mercury Toxicity to Medaka (Oryzias Latipes).
Impacts of an environmental matrix on mercury (Hg) bioavailability and toxicity to medaka (Oryzias latipes) were investigated in matrix-free controls and treatments with a stepwise increased environmental matrix of river water, sediments, and biofilms. Generally, river water enhanced but the presence of sediments and biofilms reduced Hg bioavailability to medaka up to 105 times, so that Hgtotal concentrations/amounts among different environmental media cannot mirror Hg availability and toxicity to medaka. On average, 12.9 and 12.4% of Hg in medaka was, respectively, methylated to methylmercury (MeHg) in matrix-free and -containing treatments, indicating no influence of the environmental matrix on Hg methylation in medaka. All oxidative stress, inflammatory injury, and malformation parameters correlated strongly and significantly with Hgtotal and MeHg concentrations in medaka, notably with steeper slopes in matrix-free controls than in matrix-containing treatments, highlighting that the environmental matrix mitigated Hg and MeHg toxicity to medaka. Moreover, oxidative stress was more strongly mitigated than inflammatory injury according to the stronger decreases of the regression line slopes from matrix-free to -containing treatments. Here, we have newly identified that the potential of the environmental matrix to decrease Hg bioavailability and mitigate Hg toxicity to fish together could buffer Hg ecotoxicity in the aquatic environment.


Eucalyptus globulus wood fractionation by autohydrolysis and organosolv delignification.
This work provides an assessment on the fractionation of Eucalyptus globulus wood by sequential stages of autohydrolysis (to cause the solubilization of hemicelluloses) and organosolv pulping (to dissolve lignin, leaving solids enriched in cellulose). With this approach, valuable products (hemicellulose-derived saccharides, sulphur-free lignin fragments and cellulosic substrates with low contents of residual hemicelluloses) are obtained in separate streams, according to the biomass refinery approach. Autohydrolysis was carried out under optimized operational conditions, and organosolv pulping was performed using uncatalyzed ethanol-water solutions. The effects of the most influential operational variables (autohydrolysis severity, delignification temperature and ethanol concentration in the organosolv stage) on solid yield, solid composition, cellulose susceptibility and recovery of the various fractions was assessed using statistical methods, which enabled the identification of the most favourable operational conditions.


A comprehensive review of advanced nasal delivery: Specially insulin and calcitonin.
Peptide drugs through nasal mucous membrane, such as insulin and calcitonin have been widely used in the medical field. There are always two sides to a coin. One side, intranasal drug delivery can imitate the secretion pattern in human body, having advantages of physiological structure and convenient use. Another side, the low permeability of nasal mucosa, protease environment and clearance effect of nasal cilia hinder the intranasal absorption of peptide drugs. Researchers have taken multiple means to achieve faster therapeutic concentration, lower management dose, and fewer side effects for better nasal preparations. To improve the peptide drugs absorption, various strategies had been explored via the nasal mucosa route. In this paper, we reviewed the achievements of 18 peptide drugs in the past decade about the perspectives of the efficacy, mechanism of enhancing intranasal absorption and safety. The most studies were insulin and calcitonin. As a result, absorption enhancers, nanoparticles (NPs) and bio-adhesive system are the most widely used. Among them, chitosan (CS), cell penetrating peptides (CPPs), tight junction modulators (TJMs), soft NPs and gel/hydrogel are the most promising strategies. Moreover, two or three strategies can be combined to prepare drug vectors. In addition, spray freeze dried (SFD), self-emulsifying nano-system (SEN), and intelligent glucose reaction drug delivery system are new research directions in the future.


[Contagion in literature].
Contagious diseases are very suitable to be incorporated in novels. The fact that contagious diseases are by definition a social phenomenon provides these diseases with a social, political, moral and religious dimension, apart from the medical one. New scientific concepts about contagion are adopted in novels, elaborated further and even used metaphorically, for instance in the metaphor of contagion by ideas.


Polymer Injectivity Test Design Using Numerical Simulation.
Polymer flooding is an enhanced oil recovery (EOR) process, which has received increasing interest in the industry. In this process, water-soluble polymers are used to increase injected water viscosity in order to improve mobility ratio and hence improve reservoir sweep. Polymer solutions are non-Newtonian fluids, i.e., their viscosities are shear dependent. Polymers may exhibit an increase in viscosity at high shear rates in porous media, which can cause injectivity loss. In contrast, at low shear rates they may observe viscosity loss and hence enhance the injectivity. Therefore, due to the complex non-Newtonian rheology of polymers, it is necessary to optimize the design of polymer injectivity tests in order to improve our understanding of the rheology behavior and enhance the design of polymer flood projects. This study has been addressing what information that can be gained from polymer injectivity tests, and how to design the test for maximizing information. The main source of information in the field is from the injection bottom-hole pressure (BHP). Simulation studies have analyzed the response of different non-Newtonian rheology on BHP with variations of rate and time. The results have shown that BHP from injectivity tests can be used to detect in-situ polymer rheology.


Infectious chorioretinitis in an immunocompetent patient: A diagnostic dilemma.
A 44-year-old male presented with a history of defective vision in the right eye for the past 5 months with the previous history of tubercular cervical lymphadenitis. On examination, right eye revealed panuveitis with dense vitritis and chorioretinitis in the superotemporal quadrant. His Mantoux test was positive (25 mm × 25 mm induration), QuantiFERON-TB Gold was test positive, aqueous aspirate was positive for Mycobacterium tuberculosis genome, negative for viruses and toxoplasma, and hence he was initiated on four-drug antitubercular therapy (ATT) with oral steroids. On follow-up, he had worsening of vitritis and intravenous methylprednisolone was given suspecting paradoxical reaction to ATT; however, a repeat AC tap was positive for toxoplasma B1 genome, IgG antitoxoplasma antibody was also positive in serum and aqueous; hence, we switched to systemic antitoxoplasma therapy. He underwent a therapeutic vitrectomy along with intravitreal clindamycin and dexamethasone for persistent vitreous membranes and vitritis. The patient responded well to the treatment with a reduction in vitritis and scarring of the lesion.


Plantar heel pain and foot loading during normal walking.
Plantar heel pain is aggravated by weight-bearing, yet limited evidence exists regarding how people with heel pain load their feet during walking. Knowledge of loading patterns in people with plantar heel pain would enhance the understanding of their foot function and assist in developing intervention strategies. Plantar pressure using the Emed-AT platform (Novel Gmbh, Germany) was collected from 198 people with plantar heel pain and 70 asymptomatic controls during normal walking. Maximum force, force-time integral, peak pressure, pressure-time integral and contact time were measured in four quadrants of the heel, the midfoot and the medial and lateral forefoot. The symptomatic group was sub-divided into equal low-pain and high-pain groups using the Foot Health Status Questionnaire pain score. Following age and body mass comparison, multivariate analyses of covariance were performed to compare the heel pain group to the controls, and the low-pain group to the high-pain group, for each loading variable. The heel pain group displayed lower maximum force beneath the heel, lower peak pressure beneath the postero-lateral heel and lower maximum force beneath the medial forefoot. Force-time integrals were lower beneath the posterior heel regions and higher at the lateral forefoot. People with heel pain also had longer midfoot and forefoot contact time. Higher pain level was associated with lower peak pressure and maximum force beneath regions of the heel. Compared to the controls, people with plantar heel pain demonstrated reduced heel loading and modified forefoot loading consistent with a strategy to offload the painful heel.


Neuronal activity in the cholinoceptive basal forebrain of freely moving narcoleptic dobermans.
Cholinergic stimulation in the basal forebrain (BF) triggers cataplexy in canine narcolepsy. Extracellular single unit recordings in the BF were carried out in freely moving narcoleptic dogs to study the neuronal mechanisms mediating cataplexy induction in the BF. Among the 64 recorded neurons, 12 were wake-active, three were slow wave sleep (SWS)-active, 17 were wake-/REM-active, 11 were REM sleep-active, three were cataplexy-active, and the other 18 were state-independent. Systemic administration of physostigmine, a cholinesterase inhibitor, induces status cataplecticus, decreases SWS and increases acetylcholine levels in the BF. Firing of most of the state-dependent neurons in the BF was significantly modified by physostigmine. Some of these neurons may thus mediate sleep stage changes or the effect on cataplexy observed after cholinergic stimulation in the BF.


Force output and energy metabolism during neuromuscular electrical stimulation: a 31P-NMR study.
The purpose of this study was to determine the acute physiologic effects of two electrical stimulation protocols commonly used for muscle rehabilitation. Surface electrodes were used to provide 12 stimulations of the calf musculature. In protocol A the duty cycle was fixed at 1:1 (10-second stimulation: 10-second rest); for protocol B it was 1:5 (10-second stimulation: 50-second rest). We continuously recorded isometric plantarflexor force in six healthy male subjects during stimulation using a load cell connected to a foot pedal ergometer. Metabolic changes in the stimulated gastrocnemius muscle were monitored in the supine position using 31P-NMR spectroscopy (Phillips 1.5 tesla NMR machine). Relative changes in phosphocreatine (PCr), inorganic phosphate (Pi), and intracellular pH (pHi) were obtained during stimulation and recovery, using a 1.5 cm RF surface antenna. Over the 12 stimulations, protocol A produced a significantly (p < 0.001), greater force decline (protocol A: 30.4 +/- 1.3%, protocol B: 13 +/- 0.8%); a significantly (p < 0.005), greater increase in Pi/PCr (protocol A: 210%, protocol B: 50%); and a significantly (p <0.001), lower pHi (protocol A: 6.8 +/- 0.16, protocol B: 7.03 +/- 0.12). We conclude that the shorter duty cycle produces more fatigue throughout the stimulation period, possibly as a result of greater intracellular acidosis and reduced availability of the high energy phosphate PCr. The clinical application of this finding relates to the selection of a stimulation protocol that maximizes strength gains in atrophic vs healthy muscle.


Preoperative Albumin-Bilirubin Grade as a Prognostic Predictor in Colorectal Cancer Patients Who Undergo Radical Resection.
The relationship between liver function and colorectal cancer without liver metastases has not been explored. Therefore, we investigated whether the preoperative albumin-bilirubin grade could predict the prognosis of patients with colorectal cancer (CRC) undergoing radical resection, and we designed a quantifiable predictive model. We retrospectively analyzed data from 284 patients with CRC who underwent radical resection in the Second Affiliated Hospital of the Wenzhou Medical University between January 2011 and January 2016. Patients were divided in two groups according to the calculated cut-off: the high albumin-bilirubin (>-2.48) grade and low albumin-bilirubin (≤-2.48) grade group. Kaplan-Meier curves were constructed to compare the overall survival (OS) between the two groups. Univariate and multivariate analyses were performed to identify the independent risk factors for postoperative complications and OS. Patients with a high albumin-bilirubin grade (n = 165, 58.1%) had a higher rate of postoperative complications (38.2% versus 17.6%, P < 0.001), especially medical (19.4% versus 6.7%, P = 0.002) and severe complications (1.7% versus 7.3%, P = 0.032). They also had a shorter OS (mean survival time, 47.6 versus 54.3 months, P = 0.005), especially patients with tumor-node-metastasis stage III (42.7 months versus 51.6 months, P = 0.036). Age ≥ 70 years (odds ratio [OR] = 2.22, P = 0.003) and high albumin-bilirubin grade (OR = 2.71, P = 0.001) were independent risk factors for postoperative complications, while age ≥ 70 years (hazard ratio [HR] = 2.65, P < 0.001), high albumin-bilirubin grade (HR = 1.81, P = 0.033), tumor-node-metastasis stage II (HR = 13.83, P = 0.010) and III (HR = 23.66, P = 0.002) were independent risk factors of OS. Preoperative albumin-bilirubin grade could predict postoperative complications (especially medical and severe complications) and OS in patients with CRC, especially in those with tumor-node-metastasis stage III.


Immunohistological and Ultrastructural Study of the Inflammatory Response to Perforated Polyimide Cortical Implants: Mechanisms Underlying Deterioration of Electrophysiological Recording Quality.
The deterioration of field potential (FP) recording quality and yield by in vivo multielectrode arrays (MEA) within days to weeks of implantation severely limits progress in basic and applied brain research. The prevailing hypothesis is that implantation of MEA platforms initiate and perpetuate inflammatory processes which culminate in the formation of scar tissue (the foreign body response, FBR) around the implant. The FBR leads to progressive degradation of the recording qualities by displacing neurons away from the electrode surfaces, increasing the resistance between neurons (current source) and the sensing pads and by reducing the neurons' excitable membrane properties and functional synaptic connectivity through the release of pro-inflammatory cytokines. Meticulous attempts to causally relate the cellular composition, cell density, and electrical properties of the FBR have failed to unequivocally correlate the deterioration of recording quality with the histological severity of the FBR. Based on confocal and electron microscope analysis of thin sections of polyimide based MEA implants along with the surrounding brain tissue at different points in time after implantation, we propose that abrupt FP amplitude attenuation occurs at the implant/brain-parenchyma junction as a result of high seal resistance insulation formed by adhering microglia to the implant surfaces. In contrast to the prevailing hypothesis, that FP decrease occurs across the encapsulating scar of the implanted MEA, this mechanism potentially explains why no correlations have been found between the dimensions and density of the FBR and the recording quality. Recognizing that the seal resistance formed by adhering-microglia to the implant constitutes a downstream element undermining extracellular FP recordings, suggests that approaches to mitigate the formation of the insulating glial could lead to improved recording quality and yield.


[P elements comprising mini-white marker gene suppression features in intergenic regions of Drosophila melanogaster genome].
12 492 intergenic regions were stratified into four classes according to protein coding genes mutual orientation flanking an intergenic region. It was revealed that transposon insertion sites number linearly correlates with number of promoters (none, one, or two) in an intergenic region. The vast majority oftransposons reside in intergenic regions with two promoters. Remarkably, the suppression manifestation, on the contrary, was most pronounced in ("promoterless" intergenic regions. Discussion of the phenomenon is based on the chromatin state analysis of various intergenic regions.


[Late diagnosis of Turner syndrome: therapeutic implications].
We report the case of a patient with Turner's syndrome, whose special peculiarity is that its clinical signs have gone unnoticed despite the presence of several morphological features and functional disorders which must have induced it precociously. It's about a 58 years old patient with an aortic metalic valve in treatment with dicumarinics, who came to emergencies with a severe anemic syndrome because of a peptic ulcer propitiated by the hypoprotrombinaemia. In the physical examination some typical signs of the syndrome were detected. The karyotype verified the existance of Turner (45X0). We insist on the need of an early diagnosis, in order to prevent the short stature and to offer a satisfactory sexual development, and we include an actualization of the different approachments in the therapy and management of the disease.


Juvenile versus maturity-onset alopecia areata--a comparative retrospective clinical study.
We report a retrospective study of 209 patients presenting with alopecia areata (AA) at our skin department during the period 1969-1987, with special reference to possible associations, aetiological factors and the relevance of additional investigations. The patients were divided into two groups: (I) those in whom AA developed during childhood; (II) those in whom AA developed in adult life (greater than or equal to 16 years). The aim of this study was to establish whether there is a difference between juvenile and maturity-onset AA and to consider the value of additional investigations in AA. Juvenile AA is more severe and has a less favourable prognosis than the maturity-onset disease. Statistically significant differences between the two groups were not found with respect to AA type and prevalence of auto-antibodies. Bad prognostic signs in AA were early age of onset, atopy (or first-degree family history) and ophiasis and/or onychodystrophy. Initial thyroid function testing seems advisable in patients with AA. Additional valuable investigations may include hair-root examination, determination of antibodies against thyroid tissue, and serum zinc levels. There are indications that psychosomatic factors may play a role in AA. In our study, psychosomatic factors were found in 29% of the juvenile AA and in 17% of the maturity-onset AA patients.


Inquilinus ginsengisoli sp. nov., isolated from soil of a ginseng field.
A Gram-reaction-negative, chemo-organotrophic, non-motile, non-spore-forming, rod-shaped bacterium (strain Gsoil 080(T)) was isolated from soil collected in a ginseng field in Pocheon Province, South Korea, and was investigated by using a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence analysis showed that strain Gsoil 080(T) was related most closely to Inquilinus limosus strains AU0476(T) and AU1979 (98.9 % similarity to both). Strain Gsoil 080(T) shared ≤91.3 % 16S rRNA gene sequence similarity with the type strains of other recognized species examined. The genus Inquilinus belongs to the family Rhodospirillaceae in the order Rhodospirillales, class Alphaproteobacteria. The predominant ubiquinone was Q-10 and the major fatty acids were summed feature 7 (C(18 : 1)ω9c/ω12t/ω7c) and C(19 : 0) cyclo ω8c. The G+C content of the genomic DNA of strain Gsoil 080(T) was 69.9 mol%. The level of DNA-DNA relatedness between strain Gsoil 080(T) and I. limosus LMG 20952(T) was 12 %. The results of genotypic analyses in combination with chemotaxonomic and physiological data demonstrated that strain Gsoil 080(T) represents a novel species of the genus Inquilinus, for which the name Inquilinus ginsengisoli sp. nov. is proposed. The type strain is Gsoil 080(T) (=KCTC 12574(T) =LMG 23638(T)).


Postobstructive pneumonia in lung cancer.
Postobstructive pneumonia can complicate lung cancer, particularly in more advanced stages of the disease, producing significant clinical decline and a poorer prognosis. It can lead to complications such as empyema, lung abscess and fistula formation. Postobstructive pneumonia can also be the first manifestation of an underlying malignancy. There are multiple challenges in the management of these patients. Recognition and treatment of this entity can be complex and includes the use of imaging, administration of broad-spectrum antibiotics to cover the wide variety of microorganisms involved and the use of different interventional modalities to relieve the obstruction. Existing literature on postobstructive pneumonia is scarce. In this article, we review the pathophysiology, different diagnostic methods and the therapeutic options to treat this condition. The utility and efficacy of the various modalities that are currently available in clinical practice to the interventional pulmonologist are described in some detail.


Human Serum and Salivary Metabolomes: Diversity and Closeness.
Saliva, which contains molecular information that may reflect an individual's health status, has become a valuable tool for discovering biomarkers of oral and general diseases. Due to the high vascularization of the salivary glands, there is a molecular exchange between blood and saliva. However, the composition of saliva is complex and influenced by multiple factors. This study aimed to investigate the possible relationships between the salivary and serum metabolomes to gain a comprehensive view of the metabolic phenotype under physiological conditions. Using 1H-NMR spectroscopy, we obtained the serum metabolite profiles of 20 healthy young individuals and compared them with the metabolomes of parotid, submandibular/sublingual, and whole-saliva samples collected concurrently from the same individuals using multivariate and univariate statistical analysis. Our results show that serum is more concentrated and less variable for most of the shared metabolites than the three saliva types. While we found moderate to strong correlations between serum and saliva concentrations of specific metabolites, saliva is not simply an ultrafiltrate of blood. The intense oral metabolism prevents very strong correlations between serum and salivary concentrations. This study contributes to a better understanding of salivary metabolic composition, which is crucial for utilizing saliva in laboratory diagnostics.


Impact of COVID-19 epidemic on temporal pattern of new HIV diagnoses in Italy, 2021 database.
New HIV diagnoses in Italy decreased drastically in 2020 due to COVID-19 related effects: 50% fewer diagnoses were reported by the National HIV Surveillance System. COVID-19 pandemic impact on HIV surveillance is unclear. We estimated the expected number of new HIV diagnoses in 2020 in order to isolate the impact of the COVID-19 pandemic. We analyzed 29 697 new HIV infections diagnosed from 2012 to 2020, reported to the National HIV Surveillance System. We assessed temporal trends of new HIV diagnoses applying negative binomial mixed effects models. We estimated the COVID-19 impact as the difference between the model-estimated slopes from 2012 to 2019 and the change reported in the diagnoses. The expected number of new HIV diagnoses in 2020 was also estimated and compared with the reported count. Based on the historical trend, we expected a 15% (95% CI: 5-25%) decline of new HIV diagnoses in 2020. We reported, however, a 49% decrease, yielding to a 34% net decrease in the number of new diagnoses. The strongest impact was estimated in northern regions (-40%) and MSM (-38%). We estimated 761 (95% prediction interval: 350-1277) missed diagnoses during 2020, the majority of them occurring in the North (465 cases), among MSM (416) and heterosexual males (217). In 2020, when excluding 15% decrease of new diagnoses attributable to the expected reduction, an additional 34% decrease was observed, representing a large decline in new HIV diagnoses associated with the COVID-19 pandemic.


Characterisation of chondrogenic differentiation of human mesenchymal stem cells using synchrotron FTIR microspectroscopy.
A major limiting factor in stem cell therapy is the accurate identification of the differentiation state of cells destined for transplantation. This study aimed to evaluate the potential of synchrotron radiation Fourier transform infrared (SR-FTIR) microspectroscopy as a novel technique to probe the differentiation state of human mesenchymal stem cells (hMSCs) to chondrocytes over a period of 7, 14 and 21 days of induction. The chondrogenic markers were determined using reverse transcription polymerase chain reaction, histology and immunohistochemistry. The changes of average spectra located near 1338-1230 and 1175-960 cm(-1) indicated increased levels of collagen and aggrecan, respectively, in chondrocyte-induced hMSCs compared with control cells. Classification of independent test spectra using partial least squares discriminant analysis (PLS-DA) could distinguish control and chondrocyte-induced cells with 100% accuracy. We conclude that the SR-FTIR microspectroscopy technique is sensitive for monitoring the differentiation state of stem cells under chondrogenic induction particularly at an early stage. It provides biochemical information that is complimentary to that obtained from conventional techniques, and may give more unambiguous results particularly at the very early stage of cellular differentiation. In addition, the spectroscopic approach is more straightforward, non-destructive and requires less sample preparation compared with the conventional methodologies.


Peak contact stress in human hip during gait.
The contact stress in a human hip is not uniform and it changes with different body positions. The changing location of the peak contact stress during gait may indicate the predilection sites for further development of osteoarthritis in the hip. On the basis of laboratory measurements and by using mathematical models of forces and stresses in human hip we determined the points of the peak contact stress in successive phases of gait. Results show that the peak stress points are mostly located in the posterior-medial portion of the weight bearing area, which corresponds well to the clinical observations. It is also shown that in the pathological conditions of hip dysplasia the peak contact stress trajectory is located more laterally and anteriorly.


Health care resource utilisation in primary care prior to and after a diagnosis of Alzheimer's disease: a retrospective, matched case-control study in the United Kingdom.
This study examined medical resource utilisation patterns in the United Kingdom (UK) prior to and following Alzheimer's disease (AD) diagnosis. A patient cohort aged 65 years and older with newly diagnosed AD between January 2008 and December 2010 was identified through the UK's Clinical Practice Research Datalink (CPRD). Patients with a continuous record in the CPRD (formerly the General Practice Research Database [GPRD]) for both the 3 years prior to, and the 1 year following, AD diagnosis were eligible for inclusion. A control cohort was identified by matching general older adult (GOA) patients to patients with AD based on year of birth, gender, region, and Charlson Comorbidity Index at a ratio of 2:1. Medical resource utilisation was calculated in 6-month intervals over the 4-year study period. Comparisons between AD and GOA control cohorts were conducted using conditional logistic regression for patient characteristics and a generalised linear model for resource utilisation. Data for the AD cohort (N = 3,896) and matched GOA control cohort (N = 7,792) were extracted from the CPRD. The groups were 65% female and the AD cohort had a mean age of 79.9 years (standard deviation 6.5 years) at the date of diagnosis. Over the entire study period, the AD cohort had a significantly higher mean primary care consultation rate than the GOA cohort (p < .0001). While the GOA cohort primary care consultation rate gradually increased over the 4-year period (ranging from 5 to 7 consultations per 6-month period), increases were more pronounced in the AD cohort (ranging from 6 to 11 consultations per 6-month period, peaking during the 6-month periods immediately prior to and post diagnosis). The AD cohort also had a higher overall specialty referral rate than the GOA cohort over the 4-year period (37% vs. 25%, respectively; p < .0001); the largest difference was during the 6 months immediately prior to AD diagnosis (17% vs. 5%, respectively; p < .0001). In the UK, AD diagnosis is associated with significant increases in primary and secondary care resource utilisation, continuing beyond diagnosis. This evidence may be important to health care commissioners to facilitate effective mobilisation of appropriate AD-related health care resources.


A Critical View on In Vitro Analysis of P-glycoprotein (P-gp) Transport Kinetics.
Transport proteins expressed in the different barriers of the human body can have great implications on absorption, distribution, and excretion of drug compounds. Inhibition or saturation of a transporter can potentially alter these absorbtion, distribution, metabolism and elimination properties and thereby also the pharmacokinetic profile and bioavailability of drug compounds. P-glycoprotein (P-gp, ABCB1) is an efflux transporter which is present in most of the barriers of the body, including the small intestine, the blood-brain barrier, the liver, and the kidney. In all these tissues, P-gp may mediate efflux of drug compounds and may also be a potential site for drug-drug interactions. Consequently, there is a need to be able to predict the saturation and inhibition of P-gp and other transporters in vivo. For this purpose, Michaelis-Menten steady-state analysis has been applied to estimate kinetic parameters, such as Km and Vmax, for carrier-mediated transport, whereas half-maximal inhibitor concentration (IC50) and the disassociation constant for an inhibitor/P-gp complex (Ki) have been determined to estimate P-gp inhibition. This review addresses in vitro methods commonly used to study P-gp transport kinetics and aims at providing a critical evaluation of the application of steady-state Michaelis-Menten analysis of kinetic parameters for substrate/P-gp interactions.


Preferences for cardiopulmonary resuscitation.
To examine nurse-patient communication about preferences for cardiopulmonary resuscitation (CPR). Prospective cohort. Sampled were patients and nurses caring for patients enrolled in SUPPORT (1989-91), a multicenter study of seriously-ill hospitalized adults at four U.S. hospitals. Information about patient preferences was obtained by interviews with patients and their designated surrogates. For selected patients, nurses were interviewed prospectively about their understanding of patients' preferences and whether they discussed these preferences with their patients. Nurse demographic information was obtained by questionnaire. Additional patient data were obtained by interview and chart review. Logistic regression was used to identify independent correlates of nurse-patient communication and nurses' understanding of patients' preferences. For 1,763 study patients, 1,427 nurse interviews (response rate 81%) were obtained. The median age of interviewed nurses was 29 years; 96% were women, 68% had a bachelor's or master's degree, and 62% had worked for 5 years or more as a nurse. Nurses reported discussions about CPR with 13% of their patients, and these discussions were more likely if the nurse thought the patient did not want CPR (adjusted odds ratio [AOR] 2.68; 95% CI 1.84 to 3.90), if the nurse had spent more time with the patient (AOR 1.05; 95% CI 1.02 to 1.08) per 5 additional days, if the patient had metastatic cancer (AOR 3.56; 95% CI 1.86 to 6.78), or if the patient was in an intensive care unit at the time of study entry (AOR 2.08; 95% CI 1.26 to 3.42). Diagnosis and study site were also associated with nurses' reports of discussions with patients. Of 551 patients with available data, 58% (n = 317) wanted CPR and 30% (n = 164) did not. Nurses understood patients' CPR preferences correctly for 74% of the patients. Nurses were more likely to understand patients' preferences to forego CPR if the patient was 75 years of age or older (AOR 6.6; 95% CI 2.0 to 22.0) or if the nurse and patient had discussed the patient's preferences (AOR 25.3; 95% CI 6.5 to 98.6) or if the patient had cancer (AOR 10.9; 95% CI 2.3 to 50.1). Nurses' understanding of patients' preferences for CPR was no better than that of physicians or patients' surrogate decision-makers. In this sample of seriously ill hospitalized adults, discussions between patients and nurses about CPR were infrequent. Nurses' understanding of patients' preferences for care was similar to that of physicians and patients' surrogate decision-makers. Educational interventions should focus on increasing the frequency of nurse-patient discussions about end-of-life care and improving nurses' understanding of patients' preferences for care.


Hypophysectomy abolishes rhythms in rat thyroid hormones but not in the thyroid clock.
The endocrine body rhythms including the hypothalamic-pituitary-thyroid axis seem to be regulated by the circadian timing system, and daily rhythmicity of circulating thyroid-stimulating hormone (TSH) is well established. The circadian rhythms are generated by endogenous clocks in the central brain oscillator located in the hypothalamic suprachiasmatic nucleus (SCN) as well as multiple peripheral clocks, but information on the existence and function of a thyroid clock is limited. The molecular machinery in all clock cells is composed of a number of clock genes and their gene products are connected by autoregulatory feedback loops. Here, we provide evidence for a thyroid clock in the rat by demonstrating 24-h antiphase oscillations for the mRNA of the canonical clock genes Per1 and Bmal1, which was unaffected by hypophysectomy. By immunostaining, we supported the existence of a core oscillator in the individual thyroid cells by demonstrating a daily cytoplasmatic-nuclear shuttling of PER1 protein. In normal rats, we found a significant daily rhythmicity in the circulating thyroid hormones preceded by a peak in TSH. In hypophysectomised rats, although the thyroid clock was not affected, the oscillations in circulating thyroid hormones were abolished and the levels were markedly lowered. No daily oscillations in the expression of TSH receptor mRNA were observed in neither control rats nor hypophysectomised rats. Our findings indicate that the daily rhythm of thyroid hormone secretion is governed by SCN signalling via the rhythmic TSH secretion rather than by the local thyroid clock, which was still ticking after hypophysectomy.


Determination of Methyldopa and Paracetamol in Pharmaceutical Samples by a Low Cost Genipa americana L. Polyphenol Oxidase Based Biosensor.
Purpose: Jenipapo fruit (Genipa americana L) is a natural source of polyphenol oxidases (PPOs) whose potential in pharmaceutical analysis is noteworthy. Henceforth, this work reports the electrochemical study of a low-cost PPO-based biosensor produced from the crude extract of Jenipapo fruits and accounts a practical approach to employ this biosensor in the determination of methyldopa and paracetamol in pharmaceutical samples. Methods: In order to investigate the electrochemical properties of the biosensor, theoretical and practical approaches were employed, and both samples and the biosensor were analyzed through electrochemical impedance spectroscopy (EIS) and voltammetric techniques, namely: differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Results: showcased that the biosensor presented good analytical features, as well as low detection limits (8 μmol L-1 for methyldopa and 5 μmol L-1 for paracetamol). The relative standard deviation was less than 5% mid-assay. Conclusion: The use of this biosensor is a reliable, low cost and useful alternative in the pharmaceutic determination of phenolic drugs (e.g. methyldopa and paracetamol).


Well-child care clinical practice redesign at a community health center: provider and staff perspectives.
Community health centers (CHCs) are a key element of the health care safety net for underserved children. They may be an ideal setting to create well-child care (WCC) clinical practice redesign to drastically improve WCC delivery. To examine the perspectives of clinical and administrative staff at a large, multisite urban CHC on alternative ways to deliver WCC services for low-income children aged 0 to 3 years. Eight semistructured interviews were conducted with 4 pediatric teams (each consisting of 1 pediatrician and 2 medical assistants) and 4 CHC executive/administrative staff (Medical Director, COO, CEO, and Nurse Supervisor). Discussions were recorded, transcribed, and analyzed using the constant comparative method of qualitative analysis. Salient themes included WCC delivery challenges and endorsed WCC clinical practice redesign solutions. The 3 main WCC delivery challenges included long wait times due to insurance verification and intake paperwork, lack of time for parent education and sick visits due to WCC visit volume, and absence of a system to encourage physicians to use non-face-to-face communication with parents. To address WCC delivery challenges, CHC providers and administrators endorsed several options for clinical practice redesign in their setting. These included use of a health educator in a team-based model of care, a previsit tool for screening and surveillance, Web site health education, a structured system for non-face-to-face (eg, phone) parent communication, and group visits. CHC-specific strategies for WCC clinical practice redesign endorsed by a large, multisite safety net clinic may lead to more efficient, effective, and family-centered WCC for low-income populations.


Filling Potholes on the Road to Universal Health Coverage in the Philippines.
The fraction of health-care costs financed from prepayment sources is a critical indicator of progress toward Universal Health Coverage. But it does not tell how prepayment varies with the level of health-care costs and between poorer and richer patients. This paper used survey data from the Philippines to estimate inpatient costs paid by the National Health Insurance Program (aka PhilHealth) in 2013-2017 when attempts were made to extend population, service and financial coverage. The mean fraction of the inpatient bill paid by PhilHealth increased by 21 percentage points. Expansions of population coverage do not appear to have been primarily responsible for this increase. Despite the introduction of a catastrophic cover benefit package, the fraction of inpatient costs that were prepaid increased more at lower costs than at higher costs. PhilHealth payments for inpatient care were pro-rich but became substantially less so, possibly because hospitals were no longer permitted to charge poor patients in excess of reimbursement ceilings. Overall, prepayment of inpatient costs increased and became more pro-poor, reflecting gains in insurance and equity.


Cdc25A activity is required for the metaphase II arrest in mouse oocytes.
Mammalian oocytes are arrested in metaphase of second meiosis (MII) until fertilization. This arrest is enforced by the cytostatic factor (CSF), which maintains the M-phase promoting factor (MPF) in a highly active state. Although the continuous synthesis and degradation of cyclin B to maintain the CSF-mediated MII arrest is well established, it is unknown whether cyclin-dependent kinase 1 (Cdk1) phosphorylations are involved in this arrest in mouse oocytes. Here, we show that a dynamic equilibrium of Cdk1 phosphorylation is required to maintain MII arrest. When the Cdc25A phosphatase is downregulated, mouse oocytes are released from MII arrest and MPF becomes inactivated. This inactivation occurs in the absence of cyclin B degradation and is dependent on Wee1B-mediated phosphorylation of Cdk1. Thus, our data demonstrate that Cdk1 activity is maintained during MII arrest not only by cyclin turnover but also by steady state phosphorylation.


Can plateletcrit, an underestimated platelet parameter, be related with preterm labour?
Noninvasive and simple markers are needed for the prediction of preterm delivery in women at risk for preterm labour. The aim of this study was to determine the value of platelet indices in the prediction of preterm delivery. A retrospective study. Routine antenatal care in Zonguldak Bülent Ecevit University between 2008 and 2011. Ninety patients who delivered between 28 and 37 weeks of gestational age and 128 patients who delivered at term. Plateletcrit and other haematological markers, cervical dilatation and effacement, and the neutrophil-to-lymphocyte ratio as an inflammation marker. The role of platelet indices in predicting the preterm delivery. The platelet count, plateletcrit, white blood cell count, red cell distribution width, and neutrophil count were significantly higher in the preterm delivery group. Receiver operating characteristic curve analysis showed that the plateletcrit cut-off value for predicting spontaneous preterm labour was 0.201%, with a sensitivity of 95.6% and specificity of 87.5%; the cut-off value for the platelet count was 234 ? 103/mm3 with a sensitivity of 81.0% and specificity of 71.0%. Plateletcrit is a low-cost, widely available, and noninvasive marker that might be used for the prediction of preterm delivery in patients with a history of preterm labour.


The chemokine system as a therapeutic target in autoimmune thyroid diseases: a focus on the interferon-γ inducible chemokines and their receptor.
The recruitment, trafficking, and in situ maintenance of specific subsets of activated lymphocytes constitute crucial steps for the initiation and perpetuation of chronic autoimmune inflammation. The fact that, after IFN-γ stimulation, thyrocytes secrete CXCR3- binding chemokines, which in turn recruits Th1 lymphocytes expressing CXCR3 and secreting IFN-γ, strongly supports the concept that the interferon-γ inducible chemokines (CXCL9, CXCL10, and CXCL11) and their receptor CXCR3 play an important role in the initiation of autoimmune thyroid diseases (AITD). Thus, interfering with CXCR3 might result in the abrogation of the inflammatory process. The understanding of these pathogenetic mechanisms suggested novel therapeutic approaches, with a growing interest for finding a way to interrupt the interactions between chemokines and their receptors. In this review, we focus on the efforts performed in establishing new pharmacological compounds able to target the chemokine/chemokine receptors system as well as to prevent the secretion of CXCR3-binding chemokines, induced by pro-inflammatory cytokines. The crucial issue of selecting the relevant therapeutic targets in animal models of AITD was also discussed. Although some encouraging results have been reached, major hurdles were encountered on the way to success. As a result, we are still waiting for the first anti-chemokine anti-inflammatory drug. Given the importance of leukocytes recruitment to inflammatory sites, research will continue to address the issue of developing specific chemokine-receptor antagonists. We look forward to the development of these novel pharmacological compounds which will hopefully provide a more valid alternative to the currently used lifelong replacement therapies for AITD.


Peripheral neuropathy due to vitamin deficiency, toxins, and medications.
Peripheral neuropathies secondary to vitamin deficiencies, medications, or toxins are frequently considered but can be difficult to definitively diagnose. Accurate diagnosis is important since these conditions are often treatable and preventable. This article reviews the key features of different types of neuropathies caused by these etiologies and provides a comprehensive list of specific agents that must be kept in mind. While most agents that cause peripheral neuropathy have been known for years, newly developed medications that cause peripheral neuropathy are discussed. Peripheral nerves are susceptible to damage by a wide array of toxins, medications, and vitamin deficiencies. It is important to consider these etiologies when approaching patients with a variety of neuropathic presentations; additionally, etiologic clues may be provided by other systemic symptoms. While length-dependent sensorimotor axonal peripheral neuropathy is the most common presentation, several examples present in a subacute severe fashion, mimicking Guillain-Barré syndrome.


Estimation of crash type frequencies on individual collector roadway segments.
Individual collision types have different underlying causes and thus the relationships between roadway/traffic characteristics and crash frequency are likely to differ across unique collision types. One way these different influences have been studied is by developing separate statistical models for each collision type. While this is the most straightforward approach, developing collision-specific models can be very tedious and can produce unreliable estimates for collision types that are less frequently observed. Moreover, ignoring correlations between different collision types may result in biased and inefficient parameter estimation. To overcome these limitations, researchers have adopted a multivariate approach that explicitly accounts for the correlation among individual collision types. As an alternative to multivariate approach, two-stage approaches have been proposed in which one model is estimated to predict total crash frequency and its prediction is combined with another model, used to predict the proportions of different collision types. More efficient one-stage joint models, in which both the frequency and proportion models are estimated simultaneously and predictions are provided more directly, have also been proposed for macro-level analysis. This study investigates the performance of this joint model paradigm in analyzing unique collision type frequencies on individual road segments. For this, a joint negative binomial-multinomial fractional split (NB-MFS) model is used. Moreover, this study also proposes the use of a multinomial logit (MNL) model to estimate the proportion of different collision types. As total crash frequency NB model and MNL model utilize different datasets, a two-stage estimation process is required, which leads to the two-stage NB-MNL model proposed here. The performance of proposed model is compared with that of collision-specific NB models, multivariate negative binomial (MVNB) model, and NB-MFS model in predicting crash frequency by collision type on two-way two-lane urban-suburban collector roadway segments in Pennsylvania. The goodness of fit statistics show that the NB-MNL model performs better than collision-specific NB models, MVNB model and joint NB-MFS model and is thus a promising approach in predicting crash frequency by collision type.


[Tissue plasminogen activator and plasminogen activator inhibitor gene polymorphisms: possible relation to atherosclerosis and its complications].
In the present review the dates about association of genetic variants of a plasminogen activator inhibitor type 1 (PAI-1) and tissue plasminogen activator (tPA) with ischemic heart disease and atherosclerosis of other localization are toted. The singularities of synthesis of PAI-1 and tPA, possible paths of regulation of their activity briefly surveyed. Known polymorphisms of PAI-1 gene and tPA gene, their localization, frequencies in different ethnic groups are circumscribed. In operation the genetic influence of polymorphisms of PAI-1 gene and tPA gene on synthesis and plasma concentration of PAI-1 and tPA, including under an operation of risk factors of atherosclerosis, and as possible clinical consequences of sectional violations is featured. In the review the possible role of polymorphisms of PAI-1 gene and tPA gene in a pathogenesis of atherosclerosis, association with hazard thrombotic complications for different diseases is considered. Many clinical examinations are circumscribed which have confirmed or not confirmed a possible connection of genetic markers with development and current of ischemic heart disease and other vascular diseases. We considered that there are no enough dates to one valued estimation of possible role of polymorphisms of PAI-1 and tPA genes in pathogenesis of atherosclerosis. It is necessary to get new genetic studies in groups of patient with high risk of ischemic heart disease and atherosclerosis.


Stressful Life Events Around the Time of Unplanned Pregnancy and Women's Health: Exploratory Findings from a National Sample.
Objective Little is known about how women's social context of unintended pregnancy, particularly adverse social circumstances, relates to their general health and wellbeing. We explored associations between stressful life events around the time of unintended pregnancy and physical and mental health. Methods Data are drawn from a national probability study of 1078 U.S. women aged 18-55. Our internet-based survey measured 14 different stressful life events occurring at the time of unintended pregnancy (operationalized as an additive index score), chronic disease and mental health conditions, and current health and wellbeing symptoms (standardized perceived health, depression, stress, and discrimination scales). Multivariable regression modeled relationships between stressful life events and health conditions/symptoms while controlling for sociodemographic and reproductive covariates. Results Among ever-pregnant women (N = 695), stressful life events were associated with all adverse health outcomes/symptoms in unadjusted analyses. In multivariable models, higher stressful life event scores were positively associated with chronic disease (aOR 1.21, CI 1.03-1.41) and mental health (aOR 1.42, CI 1.23-1.64) conditions, higher depression (B 0.37, CI 0.19-0.55), stress (B 0.32, CI 0.22-0.42), and discrimination (B 0.74, CI 0.45-1.04) scores, and negatively associated with ≥ very good perceived health (aOR 0.84, CI 0.73-0.97). Stressful life event effects were strongest for emotional and partner-related sub-scores. Conclusion Women with adverse social circumstances surrounding their unintended pregnancy experienced poorer health. Findings suggest that reproductive health should be considered in the broader context of women's health and wellbeing and have implications for integrated models of care that address women's family planning needs, mental and physical health, and social environments.


Surface Charge Influence on the Phase Separation and Viscosity of Cellulose Nanocrystals.
A series of four cellulose nanocrystal (CNC) suspensions were prepared from bleached softwood kraft pulp using different conditions of sulfuric acid hydrolysis. The CNCs were identical in size (95 nm in length × 5 nm in width) but had different surface charges corresponding to the harshness of the hydrolysis conditions. Consequently, it was possible to isolate the effects of surface charge on the self-assembly and viscosity of the CNC suspensions across surface charges ranging from 0.27%S to 0.89%S. The four suspensions (never-dried, free of added electrolyte) all underwent liquid crystalline phase separation, but the concentration onset for the emergence of the chiral nematic phase shifted to higher values with increasing surface charge. Similarly, suspension viscosity was also influenced by surface charge, with suspensions of lower surface charge CNCs more viscous and tending to gel at lower concentrations. The properties of the suspensions were interpreted in terms of the increase in effective diameter of the nanocrystals due to the surface electrostatic repulsion of the negative sulfate half-esters, as modified by the screening effects of the H+ counterions in the suspensions. The results suggest that there is a threshold surface charge density (∼0.3%S) above which effective volume considerations are dominant across the concentration range relevant to liquid crystalline phase formation. Above this threshold value, phase separation occurs at the same effective volume fraction of CNCs (∼10 vol %), with a corresponding increase in critical concentration due to the decrease in effective diameter that occurs with increasing surface charge. Below or near this threshold value, the formation of end-to-end aggregates may favor gelation and interfere with ordered phase formation.


A factorization approach for cone-beam reconstruction on a circular short-scan.
In this paper, we introduce a new algorithm for 3-D image reconstruction from cone-beam (CB) projections acquired along a partial circular scan. Our algorithm is based on a novel, exact factorization of the initial 3-D reconstruction problem into a set of independent 2-D inversion problems, each of which corresponds to finding the object density on one, single plane. Any such 2-D inversion problem is solved numerically using a projected steepest descent iteration scheme. We present a numerical evaluation of our factorization algorithm using computer-simulated CB data, without and with noise, of the FORBILD head phantom and of a disk phantom. First, we study quantitatively the impact of the reconstruction parameters on the algorithm performance. Next, we present reconstruction results for visual assessment of the achievable image quality and provide, for comparison, results obtained with two other state-of-the-art reconstruction algorithms for the circular short-scan.


Immune tolerance induction with recombinant factor VIII in hemophilia A patients with high responding inhibitors.
Immune tolerance induction (ITI) eradicates inhibitors in patients with hemophilia A. This study was designed to investigate the success rate of ITI in high-responding inhibitor patients with severe hemophilia A using recombinant factor VIII (rFVIII). Twenty-six patients received different ITI regimens until a normal recovery (>66%) and half-life (>6 h) of infused FVIII was achieved. In order to maximize the chance of success, the initiation of ITI was deferred in the majority of patients until the inhibitor declined to <10 BU. Twenty-two patients (85%) had baseline inhibitor levels <10 BU (median 2.3 BU) when ITI began. Within a median of 6 months, immune tolerance was achieved in 19 of 26 patients (73%) including 12/17 (70%) with intron 22 inversion, 5/7 (71%) with other null mutations and two with small deletion/insertions in the F8 gene. In conclusion, recombinant FVIII induces a high rate of immune tolerance even in carriers of null F8 mutations.


Estimation of malondialdehyde levels in serum and saliva of children affected with sickle cell anemia.
Sickle cell anemia (SCA) is an inherited disorder of hemoglobin synthesis characterized by deformed erythrocytes. Hemoglobin S present in sickle-shaped erythrocytes exhibits an enhanced rate of auto-oxidation compared with normal hemoglobin A. It produces more of reactive oxygen species (ROS) which promotes oxidatively stressed environment. ROS degrade the membranes of sickle cell erythrocytes composed of polyunsaturated lipids and form malondialdehyde (MDA) as a by-product. The aim of the study is to evaluate and compare the MDA levels of serum and saliva in SCA patients. A total of 150 children aged 4-12 years were divided into two groups: Group A (n = 75) consisting of children suffering from SCA and Group B (n = 75) consisting of healthy children. Blood and saliva samples were collected aseptically from both the groups, and they were subjected to thiobarbituric acid assay. Absorbance was evaluated spectrophotometrically at 531 nm, and the values of concentration of MDA were derived. The mean MDA levels in serum and saliva were 8.9825 ± 1.04 and 0.5152 ± 0.28, respectively, in Group A and they were found to be higher than mean MDA levels of serum (5.87 ± 0.92) and saliva (0.2929 ± 0.06) of Group B and the difference of their mean was found to be statistically significant. A significant correlation of the MDA was found in saliva and serum of the patients with SCA. This finding suggests that saliva can be effectively used as a noninvasive alternative for assessing the oxidative stress in patients with SCA.


The 'obesity paradox' in atrial fibrillation: observations from the ARISTOTLE (Apixaban for Reduction in Stroke and Other Thromboembolic Events in Atrial Fibrillation) trial.
The prognostic implication of adiposity on clinical outcomes in atrial fibrillation (AF) patients treated with oral anticoagulation is unclear. A total of 17 913 patients in the Apixaban for Reduction in Stroke and Other Thromboembolic Events in Atrial Fibrillation trial had body mass index (BMI) measured at baseline. For the primary analysis, BMI was categorized as normal (18.5 to <25 kg/m2), overweight (25 to <30 kg/m2), and obese (≥30 kg/m2). Waist circumference (WC) was defined as high if >102 cm for men and >88 cm in women. Outcomes were stroke or systemic embolism, a composite endpoint (stroke, systemic embolism, myocardial infarction, or all-cause mortality), all-cause mortality, and major bleeding. Cox models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) across categories of BMI and WC adjusting for established risk factors and treatment allocation. At baseline, 4052 (22.6%) patients had a normal BMI, 6702 (37.4%) were overweight, and 7159 (40.0%) were obese. In multivariable analyses, higher BMI was associated with a lower risk of all-cause mortality [overweight: HR 0.67 (95% CI 0.59-0.78); obese: HR 0.63 (95% CI 0.54-0.74), P < 0.0001] and the composite endpoint [overweight: HR 0.74 (95% CI 0.65-0.84); obese: HR 0.68 (95% CI 0.60-0.78), P < 0.0001] compared with normal BMI. In women, high WC was associated with a 31% lower risk of all-cause mortality (P = 0.001), 27% lower risk of the composite endpoint (P = 0.001), and 28% lower risk of stroke or systemic embolism (P = 0.048) but not in men. There was no significant association between adiposity and major bleeding. In patients with AF treated with oral anticoagulants, higher BMI and WC are associated with a more favourable prognosis.


Photoacoustic and fluorescence measurements of the chilling response and their relationship to carbon dioxide uptake in tomato plants.
The response of tomato plants to various chilling treatments was studied using two approaches for the measurement of photosynthetic activity. One involved the use of a portable fluorometer for the measurement of in-vivo chlorophyll fluorescence, while the other employed a newly introduced photoacoustic system which allowed changes in oxygen evolution to be followed in a leaf disc. A strong correlation was found between results obtained by each system and those obtained by a conventional open gas-exchange system for the determination of CO2 uptake. Both systems of measurements could readily distinguish between the effects of chilling in the dark (at 3° C for 18 h) and chilling at high photon flux density (2000 μmol m(-2) s(-1) for 5h at 5° C). Chilling in the dark had practically no effect on the quantum yield of oxygen evolution, chlorophyll fluorescence or CO2 uptake, while chilling at excessively high photon flux density resulted in a sharp reduction (50-70%) in the quantum yields obtained. The results support the view that photosystem II cannot be the primary site of damage by chilling in the dark, although it is significantly affected by chilling at high light intensity.


Identification as translation: The art of choosing the right spokespersons at the securitized border.
This article pursues a translational approach to the securitization of migration. It argues that sociotechnical processes of identification at the border can be conceived of as translations into legible identities of individuals who are unknown to authorities. The article contributes to the materiality debate on securitization across Critical Security Studies (CSS) and Science and Technology Studies (STS) by answering the call to conduct empirical explorations of security, and by revisiting the potential of the early sociology of translation (i.e. actor-network theory) to account for the identification of border crossers. Data collection was conducted at four identification facilities in the Hellenic Republic. Three sets of implications for the CSS-STS debate on the materiality of securitization are discussed. First, a translational approach can replace a representational understanding of identity with a performative apprehension of identification. Second, adopting a translational approach leads to acknowledge that the identification encounter is mediated by multiple, heterogeneous actors. It thus helps to open technological black boxes and reveal the key role of material qualities, affordances and limitations of artefacts. Third, a translational approach to the securitization of migration can help advance the field of 'alterity processing' by appreciating the de facto re-arrangements of institutional orders elicited by techno-political alignments with global security regimes.


Towards understanding of nanoparticle-protein corona.
With the rapid developments of nanotechnology, chances of exposing nanoscale particles to humans (e.g., workers and consumers) also increase correspondingly, which raises serious concerns on their biosafety. Entrance of nanoparticles into diverse biological environment endows them with new and dynamic biological identities as the so-called nanoparticle-protein corona. Therefore, understanding the role of these nanoparticle-protein coronas and resulting biological responses is crucial, as it helps to clarify the biological mechanism and prevent the potential adverse effects of nanoparticles. In this review, we summarize the latest developments relating to the nanoparticle-protein interaction and corresponding biological responses, with an emphasis on the characterization methods, induced biological effects and possible molecular mechanisms. In addition, we overview both the challenges and opportunities (particularly in nanomedicine) raised by this entrance of nanoparticles into the living creatures, especially human beings, with some future perspectives based on our understanding.


Impact of conjoined exposure to the World Trade Center attacks and to other traumatic events on the behavioral problems of preschool children.
To examine the long-term behavioral consequences of exposure to the World Trade Center (WTC) attacks in preschool children and to evaluate whether conjoined exposure to disaster and to other traumatic events has additive effects. Retrospective cohort study. Lower Manhattan, New York. A total of 116 preschool children directly exposed to the WTC attacks. Main Exposures High-intensity WTC attack-related trauma exposure indexed by the child experiencing 1 or more of the following: seeing people jumping out of the towers, seeing dead bodies, seeing injured people, witnessing the towers collapsing, and lifetime history of other trauma exposure. Main Outcome Measure Clinically significant behavioral problems as measured using the Child Behavioral Checklist. Preschool children exposed to high-intensity WTC attack-related events were at increased risk for the sleep problems and anxious/depressed behavioral symptom clusters. Conjoined exposure to high-intensity WTC attack-related events and to other trauma was associated with clinically significant emotionally reactive, anxious/depressed, and sleep-related behavioral problems. Children without a conjoined lifetime history of other trauma did not differ from nonexposed children. Risk of emotionally reactive, anxious/depressed, and attention problems in preschool children exposed to conjoined high-intensity WTC attack-related events and other trauma increased synergistically. Conjoined other trauma exposure seems to amplify the impact of high-intensity WTC attack-related events on behavioral problems. Preschool children exposed to high-intensity events who had no other trauma exposure did not have increased clinically significant behavioral problems. The additive effects of trauma exposure are consistent with an allostatic load hypothesis of stress. More vigorous outreach to trauma-exposed preschool children should become a postdisaster public health priority.


Influence of beta-naphthoflavone on 7,12-dimethylbenz(a)anthracene metabolism, DNA adduction, and tumorigenicity in rainbow trout.
Metabolism, DNA adduction, and tumor induction by 7, 12-dimethylbenz(a)anthracene (DMBA) were examined in cultured trout liver cells and in vivo in trout. Modulating CYP1A1 activity indicated this enzyme plays a significant role in metabolizing DMBA to water-soluble compounds in isolated trout liver cells. The major DMBA metabolites identified in trout liver cells were 10-, 11-, 8,9-, and 5,6-DMBA dihydrodiols, and DMBA, 2- or 3- or 4-phenol; 7-OH-methyl-12-methyl-benz(a)anthracene and 12-OH-methyl-7-methyl-benz(a)anthracene were minor metabolites. A very small amount of DMBA-3,4-dihydrodiol was detected, and polar metabolites, which did not migrate with any DMBA metabolite standards, were observed. Incubating trout hepatocytes with DMBA-3, 4-dihydrodiol produced three prominent, nonpolar adducts indistinguishable from those in mouse embryo cells. However, DMBA-DNA adducts, formed in trout in vivo or in trout liver cells exposed to DMBA, were predominantly more polar than those formed in mouse embryo fibroblasts, and levels of DMBA-DNA adducts formed in trout liver cells were not significantly altered by modulating CYP1A1 activity. No significant repair of DMBA-DNA adducts was detected in cultured trout liver cells over a 48-h period, supporting previous studies indicating that fish are less efficient than mammals in repairing polyaromatic hydrocarbon DNA adducts. Compared to animals receiving DMBA alone, beta-naphthoflavone pretreatment in vivo did not affect hepatic CYP1A1, DMBA-DNA adducts, nor hepatic tumor response; but did significantly reduce tumor response in two other target organs. These results collectively indicate that DMBA bioactivation to DNA-binding metabolites in trout liver cells and mouse embryo cells predominantly involve different metabolic pathways to form the DNA-binding intermediates.


Stability of aqueous suspensions of disulfiram.
The stability of disulfiram in aqueous suspension is reported. Aqueous suspensions of disulfiram were prepared from bulk powder and from disulfiram tablets. Acacia and sodium benzoate were present in the suspension, the final concentration of which was 25 mg/ml disulfiram. Suspensions were stored in amber bottles under fluorescent light at 24 degrees C for 178 days (tablets) and 295 days (bulk powder). Samples were taken at various intervals during storage and assayed by high-pressure liquid chromatography (HPLC) and a modified NF method. Both types of suspension were stable. Potency remained constant, and no new peaks appeared in the chromatograph. Physical integrity of the suspensions was maintained. The suspension made from tablets contained less foam and air than the one made from bulk powder; both types were easily dispersible. A degradation product of disulfiram, diethyldithiocarbamic acid, did not interfere with the assay. The HPLC assay method was more reproducible than the modified NF method. Aqueous suspensions of disulfiram are stable at room temperature for the indicated time periods, and the HPLC assay method for disulfiram is reproducible and accurate.


Reversion at the HiS1 locus of yeast.
The his1 gene (chromosome V) of Saccharomyces cerevisiae specifies phosphoribosyl transferase (E.C.18.104.22.168), the first enzyme of histidine biosynthesis. This hexameric enzyme has both catalytic and regulatory functions. The spontaneous reversion rates of seven his1 mutations were studied. The reversion rates of the alleles at the proximal end of the locus (relative to the centromere) were about 50-fold higher than distal alleles. Spontaneous reversion to prototrophy was studied in diploids homoallelic for each of the seven his1 mutations. Based on tetrad analysis, the prototrophy revertants could be assigned to three classes: (1) revertant tetrads that carried a prototrophic allele indistinguishable from wild type; (2) revertant tetrads that carried a prototrophic allele characterized by histidine excretion and feedback resistance; and (3) revertant tetrads that did not contain a prototrophic spore, but rather a newly derived allele that complemented the original allele intragenically. Four of the seven his1 mutations produced the excretor revertant class, and two mutations produced the complementer revertant class. The significance of these findings to our understanding of gene organization and the catalytic and regulatory functions of gene products are discussed.


Using infections to fight infections: paratransgenic fungi can block malaria transmission in mosquitoes.
EVALUATION OF: Fang W, Vega-Rodríguez J, Ghosh AK et al. Development of transgenic fungi that kill human malaria parasites in mosquitoes. Science 331(6020), 1074-1077 (2011). Paratransgenesis is the genetic manipulation of insect endosymbiotic microorganisms such as bacteria, viruses or fungi. Paratransgenesis has been proposed as a potential method to control vector-borne diseases such as malaria. In this article, Fang and colleagues have used genetic manipulation to insert multiple antimalaria effector genes into the entomopathogenic fungus Metarhizium anisopliae. When the modified fungus was used to infect Anopheles mosquitoes, it expressed the antimalaria effector molecules in the mosquito hemolymph. When several different effector molecules were coexpressed, malaria levels in the mosquito salivary glands were inhibited by up to 98% compared with controls. Significant inhibition could be initiated by as little as seven fungal spores and was very rapid and long lasting. These data suggest that recombinant entomopathogenic fungi could be deployed as part of a strategy to control malaria.


Platelet-type von Willebrand disease: Local disorder of the platelet GPIbα β-switch drives high-affinity binding to von Willebrand factor.
Mutations in the β-switch of GPIbα cause gain-of-function in the platelet-type von Willebrand disease. Structures of free and A1-bound GPIbα suggest that the β-switch undergoes a conformational change from a coil to a β-hairpin. Platelet-type von Willebrand disease (VWD) mutations have been proposed to stabilize the β-switch by shifting the equilibrium in favor of the β-hairpin, a hypothesis predicated on the assumption that the complex crystal structure between A1 and GPIbα is the high-affinity state. Hydrogen-deuterium exchange mass spectrometry is employed to test this hypothesis using G233V, M239V, G233V/M239V, W230L, and D235Y disease variants of GPIbα. If true, the expectation is a decrease in hydrogen-deuterium exchange within the β-switch as a result of newly formed hydrogen bonds between the β-strands of the β-hairpin. Hydrogen-exchange is enhanced, indicating that the β-switch favors the disordered loop conformation. Hydrogen-exchange is corroborated by differential scanning calorimetry, which confirms that these mutations destabilize GPIbα by allowing the β-switch to dissociate from the leucine-rich-repeat (LRR) domain. The stability of GPIbα and its A1 binding affinity, determined by surface plasmon resonance, are correlated to the extent of hydrogen exchange in the β-switch. These studies demonstrate that GPIbα with a disordered loop is binding-competent and support a mechanism in which local disorder in the β-switch exposes the LRR-domain of GPIbα enabling high-affinity interactions with the A1 domain.