Long-term intensive treatment of type 1 diabetes with the short-acting insulin analog lispro in variable combination with NPH insulin at mealtime.
To establish whether the short-acting insulin analog lispro can be successfully implemented in long-term intensive insulin therapy in type 1 diabetes, and if so, what its effects are on glycemic control and frequency and awareness of hypoglycemia. We randomized 56 type 1 diabetic patients to treatment with either lispro (n = 28) or human regular insulin (Hum-R; n = 28) as mealtime insulin for 1 year (open design, parallel groups). Lispro was injected at mealtime and Hum-R was given 10-40 min before meals (bedtime NPH was continued on both occasions). With lispro, NPH was added at breakfast (approximately 70/30), lunch (approximately 60/40), and supper (approximately 80/20) (mixing percentage of lispro/NPH) to optimize premeal and bedtime blood glucose. Total daily insulin units were no different in the two treatment groups, but with lispro approximately 30% less short-acting insulin at meals and approximately 30% more NPH was needed versus Hum-R (P < 0.05). The bedtime NPH dosage was no different. With lispro + NPH, the mean daily blood glucose was lower than with Hum-R (8.0 +/- 0.1 vs. 8.8 +/- 0.1 mmol/l; P < 0.05), HbA1c was lower (6.34 +/- 0.10 vs. 6.71 +/- 0.11%, mean value over 1 year; P < 0.002), and hypoglycemia (blood glucose < or = 3.8 mmol/l) was less frequent (7.4 +/- 0.5 vs. 11.5 +/- 0.7 episodes/patient-month) and tended to occur more within 90 min after meals than in the postabsorptive state (P < 0.05 vs. Hum-R). After 1 year, plasma adrenaline and symptom responses to experimental, stepped hypoglycemia improved with lispro and were closer to the responses of 12 nondiabetic control subjects versus Hum-R both in terms of thresholds and magnitude (P < 0.05). We concluded that mealtime injection of lispro + NPH improves the 24-h blood glucose and the percentage HbA1c as compared with Hum-R. The improvement can be maintained long term. Intensive therapy with lispro + NPH results in less frequent hypoglycemia and better awareness and counterregulation of hypoglycemia.


Retention of microbial cells in substratum surface features of micrometer and sub-micrometer dimensions.
Surfaces were produced with defined topographical features and surface chemistry. Silicon wafers, and wafers with attached nucleopore filters and quantifoils were coated with titanium using ion beam sputtering technology. Irregularly spaced, but regularly featured surface pits, sizes 0.2 and 0.5 microm, and regularly spaced pits with regular features (1 and 2 microm) diameter were produced. The smallest surface feature that could be successfully produced using this system was of diameter 0.2 microm. Ra, the average absolute deviation of the roughness irregularities from the mean line over one sampling length, Rz, the difference in height between the average of the five highest peaks, and the five lowest valleys along the assessment length of the profile and surface area values increased with surface feature size, with Ra values of 0.04-0.217 microm. There was no significant difference between the contact angles observed for smooth titanium surfaces with 0.2 and 0.5 microm features. However, a significant difference in contact angle was observed between the 1 and 2 microm featured surfaces (p<0.005). Substrata were used in microbial retention assays, using a range of unrelated, differently sized microorganisms. Staphylococcus aureus (cells 0.5-1 microm diameter) were retained in the highest numbers. S. aureus was well retained in the 0.5 microm sized pits and began to accumulate within larger surface features. Rod shaped Pseudomonas aeruginosa (1 microm x 3 microm) were preferentially retained, often end on, within the 1 microm surface features. Some daughter cells of Candida albicans blastospores were retained in 2 microm pits. For S. aureus and P. aeruginosa, the greatest numbers of cells were retained in the largest (2 microm) surface features. The number of C. albicans was similar across all the surfaces. The use of defined surfaces in microbial retention assays may lead to a better understanding of the interaction occurring between cells and surface features.


Simple few-state models reveal hidden complexity in protein folding.
Markov state models constructed from molecular dynamics simulations have recently shown success at modeling protein folding kinetics. Here we introduce two methods, flux PCCA+ (FPCCA+) and sliding constraint rate estimation (SCRE), that allow accurate rate models from protein folding simulations. We apply these techniques to fourteen massive simulation datasets generated by Anton and Folding@home. Our protocol quantitatively identifies the suitability of describing each system using two-state kinetics and predicts experimentally detectable deviations from two-state behavior. An analysis of the villin headpiece and FiP35 WW domain detects multiple native substates that are consistent with experimental data. Applying the same protocol to GTT, NTL9, and protein G suggests that some beta containing proteins can form long-lived native-like states with small register shifts. Even the simplest protein systems show folding and functional dynamics involving three or more states.


Understanding protected area resilience: a multi-scale, social-ecological approach.
Protected areas (PAs) remain central to the conservation of biodiversity. Classical PAs were conceived as areas that would be set aside to maintain a natural state with minimal human influence. However, global environmental change and growing cross-scale anthropogenic influences mean that PAs can no longer be thought of as ecological islands that function independently of the broader social-ecological system in which they are located. For PAs to be resilient (and to contribute to broader social-ecological resilience), they must be able to adapt to changing social and ecological conditions over time in a way that supports the long-term persistence of populations, communities, and ecosystems of conservation concern. We extend Ostrom's social-ecological systems framework to consider the long-term persistence of PAs, as a form of land use embedded in social-ecological systems, with important cross-scale feedbacks. Most notably, we highlight the cross-scale influences and feedbacks on PAs that exist from the local to the global scale, contextualizing PAs within multi-scale social-ecological functional landscapes. Such functional landscapes are integral to understand and manage individual PAs for long-term sustainability. We illustrate our conceptual contribution with three case studies that highlight cross-scale feedbacks and social-ecological interactions in the functioning of PAs and in relation to regional resilience. Our analysis suggests that while ecological, economic, and social processes are often directly relevant to PAs at finer scales, at broader scales, the dominant processes that shape and alter PA resilience are primarily social and economic.


Endotracheal intubation in the field improves survival in patients with severe head injury. Trauma Research and Education Foundation of San Diego.
To measure the effect of prehospital endotracheal intubation on outcome in patients with severe head injury and the percentage of these patients intubated in the field under existing protocol. Retrospective case-control study. Countywide urban trauma system. Trauma patients with blunt injury and scene Glasgow Coma Score of 8 or less, transported by ground ambulance with advanced life support capabilities from January 1, 1991, to December 31, 1995. Severe head injury was defined as head or neck Abbreviated Injury Scale score of 4 or greater. Isolated severe head injury was defined as head or neck Abbreviated Injury Scale score of 4 or greater with no other Abbreviated Injury Scale component greater than 3. One thousand ninety-two patients met initial criteria; of these, 671 had severe head injury, and 351 had isolated severe head injury. None. Mortality and functional status sufficient for discharge to home. Field intubation was associated with significant decreases in mortality from 36% to 26% in the full study group, from 57% to 36% in patients with severe head injury, and from 50% to 23% in patients with isolated severe head injury. Rate of discharge to home was unaffected by field intubation. Between 50% and 60% of study patients were intubated under current paramedic protocol, compared with intubation rates of 85% to 92% for similar patients transported by aeromedical teams operating under expanded indications for intubation. Prehospital endotracheal intubation was associated with improved survival in patients with blunt injury and scene Glasgow Coma Score of 8 or less, especially those with severe head injury by anatomic criteria. Broadening indications for intubation by paramedical personnel has great potential to improve outcome in patients with severe head injury.


Record linkage as a research tool for office-based medical care.
To explore the feasibility of linking records to study health services and health outcomes for primary care patients. A cohort of patients from the Family Medicine Centre at Mount Sinai Hospital was assembled from the clinic's billing records. Their health numbers were linked to the Ontario Hospital Discharge Database. The pattern of hospital admission rates was investigated using International Classification of Diseases (ICD) codes for primary discharge diagnosis. A pilot case-control study of risk factor management for stroke was nested in the cohort. Family medicine clinic based in a teaching hospital. A cohort of 19,654 Family Medicine Centre patients seen at least once since 1991. Admission rates by age, sex, and diagnosis. Numbers of admissions for individual patients, time to readmission, and length of stay. Odds ratios for admission for cerebrovascular disease. The 19,654 patients in the cohort had 14,299 discharges from Ontario hospitals in the 4 years from 1992 to 1995, including 3832 discharges following childbirth. Some patients had many discharges: 4816 people accounted for the 10,467 admissions excluding childbirth. Excluding transfers between institutions, there were 4975 readmissions to hospital during the 4 years, 1392 (28%) of them within 28 days of previous discharge. Admissions for mental disorders accounted for the greatest number of days in hospital. The pilot study of risk factor management suggested that acetylsalicylic acid therapy might not be effective for elderly primary care patients with atrial fibrillation and that calcium channel blocker therapy might be less effective than other therapies for preventing cerebrovascular disease in hypertensive primary care patients. Record linkage combined with data collection by chart review or interview is a useful method for studying the effectiveness of medical care in Canada and might suggest interesting hypotheses for further investigation.


Melatonin can suppress the cytotoxic effects of chlorpyrifos on human hepG2 cell lines.
The cytotoxic effect of chlorpyrifos (CP) on human HepG2 cell lines and the protective role of melatonin were investigated. TD50 of CP for HepGZ cells was also determined. The viability of HepGZ cells decreased with CP treatment in a dose-dependent manner (P <0.05). Preincubation with melatonin prior to CP application caused an increase in cell viability (P <0.05). TD50 of CP for HepG2 was determined as 84.5 microg/mL. A 1-hour melatonin treatment caused a decrease in TD50 from 84.5 to 34.1 microg/mL. The level of thiobarbituric acid reactive substance (TBARS) and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) were determined in cell lines with or without melatonin administration to find out the possible mechanism of melatonin. CP caused a significant decrease in SOD, GSH-Px and CAT activities and an increase in TBARS level (P <0.05). Pre-incubation of cells with melatonin prevented an increase in TBARS. Melatonin also reduced the CP-caused inhibition of the activities of GSH-Px and CAT (P <0.05). It was suggested that CP shows a cytotoxic effect on HepG2 cell lines and melatonin can suppress cytotoxicity caused by CP with its antioxidant properties. Melatonin also reduces TD50 of CP for HepG2 cell lines.


Discrimination between lectins with similar specificities by ratiometric profiling of binding to glycosylated surfaces; a chemical 'tongue' approach.
Carbohydrate-lectin interactions dictate a range of signalling and recognition processes in biological systems. The exploitation of these, particularly for diagnostic applications, is complicated by the inherent promiscuity of lectins along with their low affinity for individual glycans which themselves are challenging to access (bio)synthetically. Inspired by how a 'tongue' can discriminate between hundreds of flavours using a minimal set of multiplexed sensors and a training algorithm, here individual lectins are 'profiled' based on their unique binding profile (barcode) to a range of monosaccharides. By comparing the relative binding of a panel of 5 lectins to 3 monosaccharide-coated surfaces, it was possible to generate a training algorithm that enables correct identification of lectins, even those with similar glycan preferences. This is demonstrated to be useful for discrimination between the cholera and ricin toxin lectins showing the potential of this minimalist approach for exploiting glycan complexity.


Slithering snakes, angry men and out-group members: what and whom are we evolved to fear?
The preparedness theory of classical conditioning proposed by Seligman (1970, 1971) has been applied extensively over the past 40 years to explain the nature and "source" of human fear and phobias. In this review we examine the formative studies that tested the four defining characteristics of prepared learning with animal fear-relevant stimuli (typically snakes and spiders) and consider claims that fear of social stimuli, such as angry faces, or faces of racial out-group members, may also be acquired utilising the same preferential learning mechanism. Exposition of critical differences between fear learning to animal and social stimuli suggests that a single account cannot adequately explain fear learning with animal and social stimuli. We demonstrate that fear conditioned to social stimuli is less robust than fear conditioned to animal stimuli as it is susceptible to cognitive influence and propose that it may instead reflect on negative stereotypes and social norms. Thus, a theoretical model that can accommodate the influence of both biological and cultural factors is likely to have broader utility in the explanation of fear and avoidance responses than accounts based on a single mechanism.


Cyclooxygenase-2/prostaglandin D2/CRTH2 pathway mediates double-stranded RNA-induced enhancement of allergic airway inflammation.
Respiratory RNA viruses responsible for the common cold often worsen airway inflammation and bronchial responsiveness, two characteristic features of human asthma. We studied the effects of dsRNA, a nucleotide synthesized during viral replication, on airway inflammation and bronchial hyperresponsiveness in murine models of asthma. Intratracheal instillation of poly I:C, a synthetic dsRNA, increased the airway eosinophilia and enhanced bronchial hyperresponsiveness to methacholine in OVA-sensitized, exposed rats. These changes were associated with induction of cyclooxygenase-2 (COX-2) expression and COX-2-dependent PGD2 synthesis in the lungs, particularly in alveolar macrophages. The direct intratracheal instillation of PGD2 enhanced the eosinophilic inflammation in OVA-exposed animals, whereas pretreatment with a dual antagonist against the PGD2 receptor-(CRTH2) and the thromboxane A2 receptor, but not with a thromboxane A2 receptor-specific antagonist, nearly completely eliminated the dsRNA-induced worsening of airway inflammation and bronchial hyperresponsiveness. CRTH2-deficient mice had the same degree of allergen-induced airway eosinophilia as wild-type mice, but they did not exhibit a dsRNA-induced increase in eosinophil accumulation. Our data demonstrate that COX-2-dependent production of PGD2 followed by eosinophil recruitment into the airways via a CRTH2 receptor are the major pathogenetic factors responsible for the dsRNA-induced enhancement of airway inflammation and responsiveness.


Myonuclear number and myosin heavy chain expression in rat soleus single muscle fibers after spaceflight.
The effects of 14 days of spaceflight on myonuclear number, fiber size, and myosin heavy chain (MHC) expression in isolated rat soleus muscle fiber segments were studied. Single soleus muscle fibers from rats flown on the Spacelab Life Sciences-2 14-day mission were compared with those from age-matched ground-based control rats by using confocal microscopy and gel electrophoresis. Spaceflight resulted in a significant reduction in the number of fibers expressing type I MHC and an increase in the number of fibers expressing type IIx or IIa MHC. Space-flight also resulted in an increase in the percentage of fibers coexpressing more than one MHC and in the reexpression of the neonatal isoform of MHC in some fibers. Fiber cross-sectional area was significantly reduced in pure type I MHC-expressing fibers and in fibers coexpressing type I+II MHC but not in fibers expressing one or more type II MHC in the flight rats. The number of myonuclei per millimeter was significantly reduced in type I MHC-expressing fibers from the flight rats but was not significantly different in type I+II and type II MHC-coexpressing fibers. Fibers expressing neonatal MHC were similar in size to control fibers but had significantly fewer myonuclei per millimeter than flight fibers not expressing neonatal MHC. In type I MHC-expressing fibers, the reduction in fiber cross-sectional area was greater than the reduction in myonuclear number; thus the average cytoplasmic volume per myonucleus was significantly lower in flight than in control fibers. The reduction in both myonuclear number and fiber size of fibers expressing type I MHC after 14 days of spaceflight supports the hypothesis that changes in the number of myonuclei may be a contributing factor to the reduction in fiber size associated with chronic unloading of the musculature.


Compassionate Care in Behavior Analytic Treatment: Can Outcomes be Enhanced by Attending to Relationships with Caregivers?
The practice of behavior analysis has become a booming industry with growth to over 30,000 Board Certified Behavior Analysts (BCBAs) who primarily work with children with autism and their families. Most of these BCBAs are relatively novice and have likely been trained in graduate programs that focus primarily on conceptual and technical skills. Successfully working with families of children with autism, however, requires critical interpersonal skills, as well as technical skills. As practitioners strive to respond efficiently and compassionately to distressed families of children with autism, technical skills must be balanced with fluency in relationship-building skills that strengthen the commitment to treatment. The current article provides an outline of important therapeutic relationship skills that should inform the repertoire of any practicing behavior analyst, strategies to cultivate and enhance those skills, and discussion of the potential effects of relationship variables on treatment outcomes.


Involvement of linear polyubiquitylation of NEMO in NF-kappaB activation.
Nuclear factor-kappaB (NF-kappaB) is a key transcription factor in inflammatory, anti-apoptotic and immune processes. The ubiquitin pathway is crucial in regulating the NF-kappaB pathway. We have found that the LUBAC ligase complex, composed of the two RING finger proteins HOIL-1L and HOIP, conjugates a head-to-tail-linked linear polyubiquitin chain to substrates. Here, we demonstrate that LUBAC activates the canonical NF-kappaB pathway by binding to NEMO (NF-kappaB essential modulator, also called IKKgamma) and conjugates linear polyubiquitin chains onto specific Lys residues in the CC2-LZ domain of NEMO in a Ubc13-independent manner. Moreover, in HOIL-1 knockout mice and cells derived from these mice, NF-kappaB signalling induced by pro-inflammatory cytokines such as TNF-alpha and IL-1beta was suppressed, resulting in enhanced TNF-alpha-induced apoptosis in hepatocytes of HOIL-1 knockout mice. These results indicate that LUBAC is involved in the physiological regulation of the canonical NF-kappaB activation pathway through linear polyubiquitylation of NEMO.


A Mind-Body Skills Course Among Nursing and Medical Students: A Pathway for an Improved Perception of Self and the Surrounding World.
Despite increased recognition of self-care and self-awareness as core competences for health care professionals, little attention is paid to these skills during their education. Evidence suggests that a Mind-Body (MB) skills course has the potential to enhance self-care and self-awareness among medical students. However, less is known about the meaning of this course for students and how it affects their personal and professional life. Therefore, we examined the lived experiences with an MB skills course among Dutch medical and Swedish nursing students. This course included various MB techniques, such as mindfulness meditation and guided imagery. Guided by a phenomenological hermeneutical method, three main themes were identified: "ability to be more present," "increased perception and awareness of self," and "connection on a deeper level with others." Overall, participation in the MB skills course served as a pathway to inner awareness and supported connecting with others as well as with the surrounding world.


Single plane illumination fluorescence correlation spectroscopy (SPIM-FCS) probes inhomogeneous three-dimensional environments.
The life sciences require new highly sensitive imaging tools, which allow the quantitative measurement of molecular parameters within a physiological three-dimensional (3D) environment. Therefore, we combined single plane illumination microscopy (SPIM) with camera based fluorescence correlation spectroscopy (FCS). SPIM-FCS provides contiguous particle number and diffusion coefficient images with a high spatial resolution in homo- and heterogeneous 3D specimens and live zebrafish embryos. Our SPIM-FCS recorded up to 4096 spectra within 56 seconds at a laser power of 60 microW without damaging the embryo. This new FCS modality provides more measurements per time and more, less photo-toxic measurements per sample than confocal based methods. In essence, SPIM-FCS offers new opportunities to observe biomolecular interactions quantitatively and functions in a highly multiplexed manner within a physiologically relevant 3D environment.


High-Energy-Density Hydrogen-Ion-Rocking-Chair Hybrid Supercapacitors Based on Ti3C2 T x MXene and Carbon Nanotubes Mediated by Redox Active Molecule.
MXenes have emerged as promising high-volumetric-capacitance supercapacitor electrode materials, whereas their voltage windows are not wide. This disadvantage prevents MXenes from being made into aqueous symmetric supercapacitors with high energy density. To attain high energy density, constructing asymmetric supercapacitors is a reliable design choice. Here, we propose a strategy to achieve high energy density of hydrogen ion aqueous-based hybrid supercapacitors by integrating a negative electrode of Ti3C2 T x MXene and a positive electrode of redox-active hydroquinone (HQ)/carbon nanotubes. The two electrodes are separated by a Nafion film that is proton permeable in H2SO4 electrolyte. Upon charging/discharging, hydrogen ions shuttle back and forth between the cathode and anode for charge compensation. The proton-induced high capacitance of MXene and HQ, along with complementary working voltage windows, simultaneously enhance the electrochemical performance of the device. Specifically, the hybrid supercapacitors operate in a 1.6 V voltage window and deliver a high energy density of 62 Wh kg-1, which substantially exceeds those of the state-of-the-art aqueous asymmetric supercapacitors reported so far. Additionally, the device exhibits excellent cycling stability and the all-solid-state planar hybrid supercapacitor displays exceptional flexibility and integration for bipolar cells to boost the capacitance and voltage output. These encouraging results provide the possibility of designing high-energy-density noble-metal-free asymmetric supercapacitors for practical applications.


Homozygosity for the HLA-DRB1 allele selects for extraarticular manifestations in rheumatoid arthritis.
Seropositive rheumatoid arthritis is genetically linked to a group of HLA-DRB1 alleles sharing a sequence motif within the third hypervariable region. Controversy exists over the role of the distinct allelic variants in affecting not only the risk to develop disease, but also in modifying the expression of the disease. We have stratified 81 patients according to their patterns of disease manifestations and identified the HLA-DRB1 alleles by polymerase chain reaction amplification and subsequent oligonucleotide hybridization. To identify precisely the allelic combinations at the HLA-DRB1 locus, homozygosity was confirmed by locus-specific cDNA amplification and subsequent sequencing. Our study demonstrated a high correlation of allelic combinations of disease-associated HLA-DRB1 alleles with the clinical manifestations. Characteristic genotypes were identified for patients who had progressed toward nodular disease and patients who had developed major organ involvement. Rheumatoid nodules were highly associated with a heterozygosity for two disease associated HLA-DRB1 alleles. Homozygosity for the HLA-DRB1*0401 allele was a characteristic finding for RA patients with major organ involvement. Our data suggest a role of the disease-associated sequence motif in determining severity of the disease. The finding of a codominant function of HLA-DRB1 alleles suggests that the biological function of HLA-DR molecules in thymic selection might be important in the pathogenesis of RA.


Selective use of TRAM in lipopolysaccharide (LPS) and lipoteichoic acid (LTA) induced NF-kappaB activation and cytokine production in primary human cells: TRAM is an adaptor for LPS and LTA signaling.
TLR signal via Toll-IL-1R (TIR) homology domain-containing adaptor proteins. One of these adaptors, Toll-IL-1R domain-containing adaptor inducing IFN-beta-related adaptor molecule (TRAM), has been shown to be essential for TLR4 signaling in TRAM(-/-) mice and cell lines. Previously, we showed that MyD88 or Mal dominant-negative constructs did not inhibit LPS induction of cytokines in primary human M-CSF-derived macrophages. A possible explanation was redundancy of the adaptors during LPS signaling. TRAM is a suitable candidate to compensate for these adaptors. To investigate a potential role for TRAM in LPS signaling in human M-CSF-derived macrophages, we engineered an adenoviral construct expressing dominant-negative TRAM-C117H (AdTRAMdn). Synovial fibroblasts (SF) and human umbilical endothelial cells (HUVECs) were used as a nonmyeloid comparison. AdTRAMdn inhibited LPS-induced signaling in SFs and HUVECs, reducing NF-kappaB activation and cytokine production, but did not inhibit LPS signaling in M-CSF-derived human macrophages. Further investigation of other TLR ligands showed that AdTRAMdn was also able to inhibit signaling initiated by lipoteichoic acid, a TLR2 ligand, in SFs and HUVECs and lipoteichoic acid and macrophage-activating lipopeptide 2 signaling was also inhibited in TRAM(-/-) murine embryonic fibroblasts. We conclude that TRAM is an adaptor protein for both TLR4 and TLR2/6 signaling in SFs, HUVECs, and murine embryonic fibroblasts, but cannot demonstrate a role in human macrophages.


Development of a fast and user-friendly cryopreservation protocol for sweet potato genetic resources.
Sweet potato (Ipomoea batatas) is one of the ten most important staple crops and provides a livelihood for many people around the globe. To adapt to ever-changing circumstances farmers and breeders need to have access to a broad diversity of germplasm. This study focuses on the development of a cryopreservation protocol that allows the long term storage of different sweet potato cultivars. For this, a droplet vitrification protocol was optimized, comparing several parameters; preculture method (0.3 M sucrose vs no preculture); meristem position (axillary vs apical); plant age (3 to 9 weeks); regeneration medium (MS + 2.22 µM BA, Hirai and MS); and length of loading solution treatment (20 to 360 min). Two months after cryopreservation, the regeneration rates of the meristems were compared, which resulted in significant differences for the preculture method, meristem position and loading solution. With these new insights an optimized droplet vitrification protocol was developed with the following parameters: use of 3-9 week old axillary meristems, no preculture phase, 20 min LS treatment, 30 min PVS2 treatment, exposure to liquid nitrogen by droplet vitrification, warming treatment in RS for 15 min, 1 day 0.3 M sucrose recuperation culture, 1 month MS + 2.22 µM BA followed by 1 month of MS cultures. This protocol was subsequently tested on 10 representative accessions resulting in a post cryopreservation regeneration rate of more than 40% for 70% of the tested cultivars, showing that this protocol could be implemented for a large portion of existing sweet potato collections.


Problems in diagnosis and treatment of tuberculosis infection.
Tuberculosis is still a major health problem in industrialized countries due to specific socioeconomic factors and there is the growing need of new rapid and accurate diagnostic methods, in order to achieve higher sensitivity and specificity compared to traditional methods of microscopic sputum examination and culture. Such methods, recently introduced, are nucleic acid amplification (NAA) tests, used directly on clinical specimens and blood tests (QuantiFERON-TB, T-SPOT.TB test), measuring the IFN-gamma released by stimulated T cells. Furthermore, new drugs for the disease need to be developed, aiming to better treatment results and to prevention of Multiple Drug Resistance (MDR) cases. Critical aspects in the management of drug resistance cases should be the careful choices of drugs combination, the close follow up of the patients alongside with the patients adherence to therapy. The role of national and international tuberculosis programs is invaluable in TB control and therapy, as well as the collaboration of all the health system departments. However, most of the clinical problems that may arise are addressed by the International Standards for Tuberculosis Care-ISTC and these guidelines should be taken into consideration, at least until future research provides more promising diagnostic and therapeutic modalities for control of the disease.


Indocyanine Green Angiography Predicts Tissue Necrosis More Accurately Than Thermal Imaging and Near-Infrared Spectroscopy in a Rat Perforator Flap Model.
Clinical examination alone is neither sensitive nor specific for predicting flap necrosis, so several technologies, including indocyanine green angiography, thermal imaging (using the FLIR ONE), and near-infrared spectroscopy, have been developed to supplement perfusion assessment. This study aims to compare the accuracy of these three methods for intraoperatively predicting clinical flap necrosis in a rat perforator flap model. The authors hypothesized that near-infrared spectroscopy, assessing oxygenation rather than direct perfusion, would yield significantly different predictions. A 10 × 3-cm epigastric perforator flap was elevated in 14 adult male rats weighing 250 ± 50 g. Flap perfusion was assessed immediately after flap elevation using thermal imaging, near-infrared spectroscopy, and indocyanine green angiography. Measurements were correlated to the clinical endpoint and gold standard of flap necrosis on postoperative day 7. All three technologies detected significant differences in perfusion along flap length (all p < 0.001), and were associated with significant differences in the odds of developing flap necrosis (all p < 0.001). The areas under the receiver operating characteristic curves were 0.948 for indocyanine green angiography as an absolute value, 0.873 for relative changes with thermal imaging, and 0.792 for tissue oxygenation. The sensitivity, specificity, and accuracy for indocyanine green angiography measured as an absolute value were the highest at 97.8, 87.5, and 92 percent, respectively. Indocyanine green angiography most accurately predicted flap necrosis in this study; however, tissue oximetry and thermal imaging were also capable of predicting necrosis and represented potentially less expensive or more readily available alternatives for objective perfusion assessment. Additional research can further delineate their roles and cost-efficacy in clinical practice.


Integrated optical frequency-resolved Mach-Zehnder interferometers for label-free affinity sensing.
Integrated Optical Frequency-Resolved Mach-Zehnder Interferometry (IO FR-MZI) is introduced as an alternative, cost-efficient operation principle for integrated optical label-free affinity sensors that can combine high sensitivity with high versatility in terms of potential applications and experimental configurations. A detailed theoretical analysis of the method is presented followed by a semi-analytical approximation and numerical calculations in order to quantify the sensitivity and limits of detection of the FR-MZI over Single Wavelength MZI. The obtained results substantiate that IO FR-MZI- based sensors constitute a generic technological platform of high sensitivity that can be implemented into a plethora of detection schemes. For an optimized optical design well below 1mm in length the limit of detection can be as low as 0.025A in terms of adlayer effective thickness allowing for truly miniaturized integrated optical sensors fabricated with high yield with standard microfabrication techniques.


Effect of caffeine and adenosine receptor ligands on the expression of spike-and-wave discharges in Genetic Absence Epilepsy Rats from Strasbourg (GAERS).
The influence of caffeine on epileptic seizures remains a matter of debate. Here we tested on Genetic Absence Epilepsy Rats from Strasbourg (GAERS) the consequences of acute and chronic exposure to caffeine on the expression of spike-and-wave discharges (SWDs). Since caffeine is a mixed nonspecific A(1) and A(2A) adenosine receptor antagonist, we measured also the influence of antagonists and agonists of these receptors on SWD expression. GAERS were equipped with four cortical electrodes over the frontoparietal cortex and the cumulated duration and number of SWDs were recorded for 120 min after the injection of increasing doses of caffeine, specific antagonists and agonists of A(1) and A(2A) adenosine receptors. The effects of chronic caffeine were also studied. In GAERS, caffeine dose-dependently reduced the cumulated number and duration of SWDs which almost disappeared after the injection of the two highest doses of caffeine, 5 and 10 mg/kg. Likewise, the A(1) and A(2A) adenosine receptor antagonists led to a dose-dependent reduction of SWD expression while the agonists dose-dependently increased SWD expression. Conversely, the chronic exposure to caffeine via drinking water for 15 days did not influence SWD expression. With the exception of the two highest doses of caffeine that largely enhanced activity, all compounds including low doses of caffeine had no effect on locomotor activity of GAERS. These data show that the acute exposure to low doses of caffeine, or A(1) and A(2A) adenosine receptor antagonists reduces SWD expression in GAERS, independently from any effect on motor activity. The chronic exposure of GAERS to caffeine does not affect the expression of epilepsy.


Flavin-Dependent Monooxygenase-Mediated 1,2-Oxazine Construction via Meisenheimer Rearrangement in the Biosynthesis of Paeciloxazine.
The [1,2]-Meisenheimer rearrangement is well known as the [1,2]-migration of an O-substituted hydroxylamine from a tertiary amine N-oxide, and it is frequently employed in organic synthesis to enforce adjacent carbon oxidation or install a 1,2-oxazine core, which is a prevalent structural feature and pharmacophore of many bioactive natural products. Although the [1,2]-Meisenheimer rearrangement was proposed to occur in the biosynthesis of a number of 1,2-oxazine-containing natural products, it has never been proved biosynthetically. Here, we identified the biosynthetic gene cluster of an insecticidal natural product, paeciloxazine (1), from Penicillium janthinellum and characterized a flavin-dependent monooxygenase, PaxA, as the first example that mediates the formation of a 1,2-oxazine moiety via Meisenheimer rearrangement. In vitro biochemical assays, site-directed mutations, docking and molecular dynamics simulations, and density functional theory calculations support the mechanism that PaxA first catalyzes N-oxidation to form an N-oxide intermediate, which undergoes [1,2]-Meisenheimer rearrangement with the assistance of an amino acid with proton transfer property. This study expands the repertoire of rearrangement reactions during the biosynthesis of natural products and provides a new strategy for discovering natural products with N-O tethers by genome mining.


Effect of prostaglandins on cyclic AMP production in cultured human ciliary muscle cells.
Prostaglandins (PGs) lower intraocular pressure by increasing uveoscleral outflow, presumably via a receptor-mediated mechanism coupled to a second messenger pathway in the ciliary muscle. In the present study, we examined the effect of prostanoids on cyclic AMP production in cultured human ciliary muscle cells. Cells were identified based on their expression of smooth muscle specific alpha-actin and monoclonal antibody against desmin. Cyclic AMP production in confluent cells incubated with buffer solution containing various concentrations of prostanoids was analyzed by radioimmunoassay. PGE2 caused a time-dependent increase in cyclic AMP concentrations which reached a maximum after 10 mins. With the exception of PGD2, all prostanoids produced a concentration-dependent increase in cyclic AMP levels with the following rank order of activity: PGE2 > 11-deoxy-PGE1 > 16,16-dimethyl PGE2 > sulprostone > PGF2alpha. PGE2-induced increase on cyclic AMP levels was unaffected by AH6809, an antagonist at both PGD2 (DP) and E2 (EP1) receptors. Flurbiprofen decreased basal cyclic AMP concentrations suggesting that intramurally-generated PGs stimulate the formation of the nucleotide in ciliary smooth muscle cells. PGE2-induced increases in cyclic AMP production was synergistic with those induced by the diterpene activator of adenylyl cyclase, forskolin. We conclude that prostanoids active at EP2-receptors can stimulate cyclic AMP production in cultured human ciliary muscle cells.


Regional bivalent-univalent pairing versus trivalent pairing of a trisomic chromosome in Saccharomyces cerevisiae.
In meiosis I, homologous chromosomes pair, recombine and segregate to opposite poles. These events and subsequent meiosis II ensure that each of the four meiotic products has one complete set of chromosomes. In this study, the meiotic pairing and segregation of a trisomic chromosome in a diploid (2n + 1) yeast strain was examined. We find that trivalent pairing and segregation is the favored arrangement. However, insertions near the centromere in one of the trisomic chromosomes leads to preferential pairing and segregation of the "like" centromeres of the remaining two chromosomes, suggesting that bivalent-univalent pairing and segregation is favored for this region.


Identification of a new locus, Ptr(t), required for rice blast resistance gene Pi-ta-mediated resistance.
Resistance to the blast pathogen Magnaporthe oryzae is proposed to be initiated by physical binding of a putative cytoplasmic receptor encoded by a nucleotide binding site-type resistance gene, Pi-ta, to the processed elicitor encoded by the corresponding avirulence gene AVR-Pita. Here, we report the identification of a new locus, Ptr(t), that is required for Pi-ta-mediated signal recognition. A Pi-ta-expressing susceptible mutant was identified using a genetic screen. Putative mutations at Ptr(t) do not alter recognition specificity to another resistance gene, Pi-k(s), in the Pi-ta homozygote, indicating that Ptr(t) is more likely specific to Pi-ta-mediated signal recognition. Genetic crosses of Pi-ta Ptr(t) and Pi-ta ptr(t) homozygotes suggest that Ptr(t) segregates as a single dominant nuclear gene. A ratio of 1:1 (resistant/susceptible) of a population of BC1 of Pi-ta Ptr(t) with pi-ta ptr(t) homozygotes indicates that Pi-ta and Ptr(t) are linked and cosegregate. Genotyping of mutants of pi-ta ptr(t) and Pi-ta Ptr(t) homozygotes using ten simple sequence repeat markers at the Pi-ta region determined that Pi-ta and Ptr(t) are located within a 9-megabase region and are of indica origin. Identification of Ptr(t) is a significant advancement in studying Pi-ta-mediated signal recognition and transduction.


On the linear geometry of lanthanide hydroxide (Ln-OH, Ln = La-Lu).
Lanthanide hydroxides are key species in a variety of catalytic processes and in the preparation of corresponding oxides. This work explores the fundamental structure and bonding of the simplest lanthanide hydroxide, LnOH (Ln = La-Lu), using density functional theory calculations. Interestingly, the calculations predict that all structures of this series will be linear. Furthermore, these results indicate a valence electron configuration of σ2π4 for all LnOH compounds, suggesting that the lanthanide-hydroxide bond is best characterized as a covalent triple bond.


Avian hind-limb digit length ratios measured from radiographs are sexually dimorphic.
Sexual dimorphism in digit length ratios is well established in humans, and has been reported in other vertebrate species as well, including birds. The sign of sexual dimorphism in digit ratios may, however, vary both within and between vertebrate classes. It has been hypothesized that sex differences in digit ratios arise via differential prenatal exposure of the two sexes to steroids, which may affect the expression of the Hox genes controlling the osteometric development of digits and appendices. Among birds, the evidence for sex dimorphism in hind-limb digit ratios is conflicting, though all previous studies were based on measurements of undissected digits, implying that results could be confounded by sex-related variation in soft tissues. Here we report that digit ratios derived from radiographs of both feet of a large passerine bird, the hooded crow (Corvus corone), are sexually dimorphic, males showing larger 2D : 3D (effect size, r = 0.33) and 2D : 4D than females (effect size, r = 0.28). We also observed a good agreement (r = 0.45) between radiographic estimates of digit ratios and digit ratios calculated based on undissected digit measurements (thus including soft tissues). Importantly, we found that the patterns of sex and side differences were largely coherent between the two methods. Therefore, our findings show for the first time in avian species that sex differences in digit ratios have an osteometric basis, a fundamental prerequisite for a role of Hox genes in originating such dimorphism.


Synthesis and molecular and electronic structures of reduced bis(imino)pyridine cobalt dinitrogen complexes: ligand versus metal reduction.
Sodium amalgam reduction of the aryl-substituted bis(imino)pyridine cobalt dihalide complexes ((Ar)PDI)CoCl(2) and ((iPr)BPDI)CoCl(2) ((Ar)PDI = 2,6-(2,6-R(2)-C(6)H(3)N=CMe)(2)C(5)H(3)N (R = (i)Pr, Et, Me); (iPr)BPDI = 2,6-(2,6-(i)Pr(2)-C(6)H(3)N=CPh)(2)C(5)H(3)N) in the presence of an N(2) atmosphere furnished the corresponding neutral cobalt dinitrogen complexes ((Ar)PDI)CoN(2) and ((iPr)BPDI)CoN(2). Magnetic measurements on these compounds establish doublet ground states. Two examples, ((iPr)PDI)CoN(2) and ((iPr)BPDI)CoN(2), were characterized by X-ray diffraction and exhibit metrical parameters consistent with one-electron chelate reduction and a Co(I) oxidation state. Accordingly, the toluene solution EPR spectrum of ((iPr)PDI)CoN(2) at 23 degrees C exhibits an isotropic signal with a g value of 2.003 and hyperfine coupling constant of 8 x 10(-4) cm(-1) to the I = 7/2 (59)Co center, suggesting a principally bis(imino)pyridine-based SOMO. Additional one-electron reduction of ((iPr)PDI)CoN(2) was accomplished by treatment with Na[C(10)H(8)] in THF and yielded the cobalt dinitrogen anion [((iPr)PDI)CoN(2)](-). DFT calculations on the series of cationic, neutral, and anionic bis(imino)pyridine cobalt dinitrogen compounds establish Co(I) centers in each case and a chelate-centered reduction in each of the sequential one-electron reduction steps. Frequency calculations successfully reproduce the experimentally determined N[triple bond]N infrared stretching frequencies and validate the computational methods. The electronic structures of the reduced cobalt dinitrogen complexes are evaluated in the broader context of bis(imino)pyridine base metal chemistry and the influence of the metal d electron configuration on the preference for closed-shell versus triplet diradical dianions.


Computational complexity in electronic structure.
In quantum chemistry, the price paid by all known efficient model chemistries is either the truncation of the Hilbert space or uncontrolled approximations. Theoretical computer science suggests that these restrictions are not mere shortcomings of the algorithm designers and programmers but could stem from the inherent difficulty of simulating quantum systems. Extensions of computer science and information processing exploiting quantum mechanics has led to new ways of understanding the ultimate limitations of computational power. Interestingly, this perspective helps us understand widely used model chemistries in a new light. In this article, the fundamentals of computational complexity will be reviewed and motivated from the vantage point of chemistry. Then recent results from the computational complexity literature regarding common model chemistries including Hartree-Fock and density functional theory are discussed.


Pelvis Anatomy Tutorial Using Radiological Images.
Improving integration between basic science and clinical application is essential in medical education. Anatomy courses can do this by focusing on medical imaging interpretation. Most imaging textbooks rely on structural identification, which novice learners often struggle to apply to the health care environment, particularly in complex regions like the pelvis, which is multifaceted and differs substantially between sexes. To address this deficit, this resource extends our imaging-based tutorial series. This tutorial was a self-administered PowerPoint incorporating X-ray, computed tomography, and magnetic resonance imaging, which are all often used for the pelvic region, as well as self-quizzing and clinical applications. Using repeated-measures, control/experimental design, the tutorial was evaluated as a review tool for 57 Australian medical students in preclerkship years. Participants were evaluated by a rating self-efficacy scale, knowledge-based testing (multiple-choice, short-answer, and identification questions), and feedback to open-ended questions. Results indicate that the tutorial significantly improved direct knowledge (p = .006), as the experimental group's posttutorial scores for direct questions were superior by 21% on average. Significant improvements occurred specifically for direct short-answer and indirect image-identification questions. These results suggest the tutorial is an effective review tool. While previous tutorials were evaluated as adjunct tools, this tutorial was evaluated post-anatomy teaching with similar results. Students improved in direct and applied anatomy following tutorial exposure. This suggests that the tutorial series comprises valuable review and supplementary materials. None of our tutorials have been evaluated as a sole mechanism for teaching anatomy or imaging.


Uncovering the unexplored diversity of thioamidated ribosomal peptides in Actinobacteria using the RiPPER genome mining tool.
The rational discovery of new specialized metabolites by genome mining represents a very promising strategy in the quest for new bioactive molecules. Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a major class of natural product that derive from genetically encoded precursor peptides. However, RiPP gene clusters are particularly refractory to reliable bioinformatic predictions due to the absence of a common biosynthetic feature across all pathways. Here, we describe RiPPER, a new tool for the family-independent identification of RiPP precursor peptides and apply this methodology to search for novel thioamidated RiPPs in Actinobacteria. Until now, thioamidation was believed to be a rare post-translational modification, which is catalyzed by a pair of proteins (YcaO and TfuA) in Archaea. In Actinobacteria, the thioviridamide-like molecules are a family of cytotoxic RiPPs that feature multiple thioamides, which are proposed to be introduced by YcaO-TfuA proteins. Using RiPPER, we show that previously undescribed RiPP gene clusters encoding YcaO and TfuA proteins are widespread in Actinobacteria and encode a highly diverse landscape of precursor peptides that are predicted to make thioamidated RiPPs. To illustrate this strategy, we describe the first rational discovery of a new structural class of thioamidated natural products, the thiovarsolins from Streptomyces varsoviensis.


Predicting localised measles outbreak potential in Australia.
Australia has achieved measles elimination as announced in March 2014 by the WHO Western Pacific Regional Committee, based on several lines of evidence. However, despite strong national evidence for elimination, there remains substantial regional variation in vaccine coverage, has resulted in recent outbreaks and potential for increased frequency in the future. In this study, we apply a multiple cohort model of measles immunity, stratified by age and local geographic area to predict trends in the measles reproduction number R. In addition, we use branching process models of outbreak risks to predict state-level probabilities of the occurrence of measles outbreaks over the next 20 years. Our results suggest increasing risks of large measles outbreaks over this period, in particular in the states of Queensland and New South Wales. In addition, there is wide variation in predicted R values by smaller geographic areas, although uncertainty in age-specific immunity limits the precision of our results. Our predictions align with observed outbreaks in Australian states and suggest our approach to determining future outbreak risks could be applied more widely in elimination or near-elimination settings.


Measures of visuospatial short-term memory: the Knox Cube Imitation Test and the Corsi Blocks Test compared.
The present investigation analyzes the characteristics of two tasks that have been considered as a measure of visuospatial abilities: The Knox Cube Imitation Test and the Corsi Blocks Test. The former was originally devised by Knox (1913) to diagnose mental retardation in potential immigrants to the United States, while the latter has been specifically designed to be used in neuropsychological practice by Corsi (1972). Although both tasks have been widely used in the past, there is little empirical research investigating the characteristics of these tasks from a theoretical point of view. In order to do so, we carried out two parallel experiments in which both tasks were presented in a baseline condition as well as in association with three different concurrent tasks (i.e., articulatory suppression, spatial tapping, and random generation) supposed to tap the various components of working memory. Results showed that neither of the tasks can be considered as a pure measure of visuospatial processing and that, at the same time, it is necessary to reconsider the architecture of working memory in order to suggest a more integrated functioning of the system.


Obstetric Care Consensus No. 2: Levels of maternal care.
In the 1970s, studies demonstrated that timely access to risk-appropriate neonatal and obstetric care could reduce perinatal mortality. Since the publication of the Toward Improving the Outcome of Pregnancy report, more than three decades ago, the conceptual framework of regionalization of care of the woman and the newborn has been gradually separated with recent focus almost entirely on the newborn. In this current document, maternal care refers to all aspects of antepartum, intrapartum, and postpartum care of the pregnant woman. The proposed classification system for levels of maternal care pertains to birth centers, basic care (level I), specialty care (level II), subspecialty care (level III), and regional perinatal health care centers (level IV). The goal of regionalized maternal care is for pregnant women at high risk to receive care in facilities that are prepared to provide the required level of specialized care, thereby reducing maternal morbidity and mortality in the United States.


Safety of artemisinins in first trimester of prospectively followed pregnancies: an observational study.
Artemisinins, the most effective antimalarials available, are not recommended for falciparum malaria during the first trimester of pregnancy because of safety concerns. Therefore, quinine is used despite its poor effectiveness. Assessing artemisinin safety requires weighing the risks of malaria and its treatment. We aimed to assess the effect of first-trimester malaria and artemisinin treatment on miscarriage and major congenital malformations. In this observational study, we assessed data from antenatal clinics on the Thai-Myanmar border between Jan 1, 1994, and Dec 31, 2013. We included women who presented to antenatal clinics during their first trimester with a viable fetus. Women were screened for malaria, and data on malaria, antimalarial treatment, and birth outcomes were collected. The relationship between artemisinin treatments (artesunate, dihydroartemisinin, or artemether) and miscarriage or malformation was assessed using Cox regression with left-truncation and time-varying exposures. Of 55 636 pregnancies registered between 1994 and 2013, 25 485 pregnancies were analysed for first-trimester malaria and miscarriage, in which 2558 (10%) had first-trimester malaria. The hazard of miscarriage increased 1·61-fold after an initial first-trimester falciparum episode (95% CI 1·32-1·97; p<0·0001), 3·24-fold following falciparum recurrence (2·24-4·68; p<0·0001), and 2·44-fold (1·01-5·88; p=0·0473) following recurrent symptomatic vivax malaria. No difference was noted in miscarriage in first-line falciparum treatments with artemisinin (n=183) versus quinine (n=842; HR 0·78 [95% CI 0·45-1·34]; p=0·3645) or in risk of major congenital malformations (two [2%] of 109 [95% CI 0·22-6·47] versus eight (1%) of 641 [0·54-2·44], respectively). First-trimester falciparum and vivax malaria both increase the risk of miscarriage. We noted no evidence of an increased risk of miscarriage or of major congenital malformations associated with first-line treatment with an artemisinin derivative compared with quinine. In view of the low efficacy of quinine and wide availability of highly effective artemisinin-based combination therapies, it is time to reconsider first-trimester antimalarial treatment recommendations. The Wellcome Trust and The Bill & Melinda Gates Foundation.


Wait times for diagnosis and treatment of lung cancer: a single-centre experience.
Multiple clinical practice guidelines recommend rapid evaluation of patients with suspected lung cancer. It is uncertain whether delays in diagnosis and management have a negative effect on outcomes. This retrospective study included 551 patients diagnosed with lung cancer through the diagnostic assessment program at the Institut universitaire de cardiologie et de pneumologie de Québec between September 2013 and March 2015. Median wait times between initial referral, diagnosis, and first treatment were calculated and compared with recommended targets. Analyses were performed to evaluate for specific factors associated with longer wait times and for the effect of delays on the outcomes of progression-free survival (pfs), relapse-free survival (rfs) after primary surgical resection, and overall survival (os). Most patients were investigated and treated within recommended targets. Of the entire cohort, 379 patients were treated at our institution. Of those 379 patients, 311 (82%) were treated within recommended targets. In comparing patients within and outside target times, the only statistically significant difference was found in the distribution of treatment modalities: patients meeting targets were more likely to be treated with surgery or chemotherapy rather than with radiation. The pfs on first treatment modality was influenced by clinical stage, but not by time to therapy [hazard ratio (hr): 1.10; p = 0.65]. The os for the entire cohort was also influenced by stage, but not by delays (hr: 1.04; p = 0.87). For the 209 patients treated by surgery with curative intent, a significant reduction in rfs was associated with male sex and TNM stage, but not with delays (hr: 1.11; p = 0.83). The os after primary surgical resection was also associated with TNM stage, but not with delays (hr: 1.82; p = 0.43). Recommended targets for wait times in the investigation and treatment of lung cancer can be achieved within a diagnostic assessment program. Compared with radiation treatment, treatment with surgery or chemotherapy is more likely to be completed within targets. Delays in investigation and treatment do not appear to negatively affect the clinical outcomes of os, rfs, and pfs. Prospective studies are needed to evaluate whether efficient work-up and treatment influence other important variables, such as quality of life, cost of care, and access to therapies while performance status is adequate.


Emergency medicine: what keeps me, what might lose me? A narrative study of consultant views in Wales.
EDs are currently under intense pressure due to increased patient demand. There are major issues with retention of senior personnel, making the specialty a less attractive choice for junior doctors. This study aims to explore what attracted EM consultants to their career and keeps them there. It is hoped this can inform recruitment strategies to increase the popularity of EM to medical students and junior doctors, many of whom have very limited EM exposure. Semistructured interviews were conducted with 10 consultants from Welsh EDs using a narrative approach. Three main themes emerged that influenced the career choice of the consultants interviewed: (1) early exposure to positive EM role models; (2) non-hierarchical team structure; (3) suitability of EM for flexible working. The main reason for consultants leaving was the pressure of work impacting on patient care. The study findings suggest that EM consultants in post are positive about their careers despite the high volume of consultant attrition. This study reinforces the need for dedicated undergraduate EM placements to stimulate interest and encourage medical student EM aspirations. Consultants identified that improving the physical working environment, including organisation, would increase their effectiveness and the attractiveness of EM as a long-term career.


Efficacy and Safety of Rituximab for the Treatment of Graves' Orbitopathy: A Meta-analysis of Randomized Controlled Trials.
To investigate the efficacy and safety of rituximab in patients with Graves' orbitopathy (GO). Systematic review and meta-analysis of four randomized controlled trials. A total of 293 patients with GO who received rituximab or control (either glucocorticoids, the established first-line therapy [three trials], or saline [one trial]). Relevant studies published before February 2018 were identified from the PubMed, EMBASE, Cochrane Library, and Scopus databases and the ClinicalTrials.gov registry. Individual effect sizes were standardized, and a meta-analysis was conducted to calculate the pooled effect size by using a random-effects model. Treatment efficacy was assessed by measuring the following outcomes: clinical activity score (CAS), sight visual acuity reduction (NOSPECS) score, proptosis, diplopia, changes in eye volume, quality of life, and adverse events. In the four included trials, 113 patients in the rituximab group and 108 patients in the control group were evaluated. Compared with the control group, CAS (weighted mean difference 0.57, 95% confidence interval 0.25-0.89) was significantly reduced at 24 weeks in the rituximab group. Compared with the control group, considerable proptosis reduction was also observed in the rituximab group; however, the difference was not significant. The proportion of adverse events in the rituximab group was not significantly higher than that in the glucocorticoid control group, but one of the included trials indicated that the rituximab group had more serious adverse events than the saline control group. Rituximab is a relatively safe and viable treatment that is superior to glucocorticoids or saline for patients with moderate to severe GO. However, the incidence of serious adverse events was disparate among the included trials. Additional studies involving a larger sample size and investigating the optimal rituximab dosage, frequency, and method of administration are warranted.


Are sore throat patients who hope for antibiotics actually asking for pain relief?
Antibiotics are still overprescribed for self-limiting upper respiratory tract infections such as acute sore throat, and physicians mention patient's desire for antibiotics as a driving force. We studied patients' concerns when visiting their family physician for acute sore throat, more specifically the importance they attach to antibiotic treatment and pain relief. Family physicians in 6 peer groups in Belgium participated in an observational postvisit questionnaire survey. Patients aged 12 years and older making an office visit for acute sore throat were invited to indicate the importance of different reasons for the visit. Sixty-eight family physicians provided data from 298 patients. The 3 most frequently endorsed reasons for visiting the physician were examination to establish the cause of the symptoms, pain relief, and information on the course of the disease. Hopes for an antibiotic ranked 11th of 13 items. Patients who considered antibiotics very/rather important valued pain relief significantly more than patients who considered them little/not important (P <.001). Patients who hoped for antibiotics felt more unwell (P <.001), had more faith in antibiotics to speed recovery (P <.001), and were less convinced that sore throat was a self-limiting disease (P <.012). A multivariate model, adjusted for age, sex, and educational status, showed that the desire for pain relief is a strong predictor of the hope to receive a prescription for antibiotics. Our study suggests that patients with acute sore throat and who hope for antibiotics may in fact want treatment for pain. Trials are needed to test whether exploring patients' expectations about pain management and offering adequate analgesia can assist physicians in managing sore throats without prescribing antibiotics.


Mild head injuries: impact of a national strategy for implementation of management guidelines.
A national survey in 1996 showed insufficient routines for management of patients with mild head injuries in Norwegian hospitals. Since then, the Scandinavian Guidelines for Management of Mild Head Injuries have been published. A cross-sectional questionnaire survey of management practice was performed in all 59 hospitals in 2002. We compared the results with figures from 1996 and evaluated guideline compliance. The proportion of noncompliant hospitals was reduced (p = 0.02) from 52% to 31%. The proportion assessing the patient's level of consciousness according to the Glasgow Coma Scale increased (p = 0.001) from 49% to 80%. The proportion requiring a normal computed tomographic scan if a patient with a history of loss of consciousness was to be sent home from the accident and emergency department increased (p < 0.001) from 1 (2%) to 13 (19%). The Scandinavian Guidelines for Management of Mild Head Injuries have had a significant impact on management practice in Norwegian hospitals.


Modulation of defensive reactivity by GLRB allelic variation: converging evidence from an intermediate phenotype approach.
Representing a phylogenetically old and very basic mechanism of inhibitory neurotransmission, glycine receptors have been implicated in the modulation of behavioral components underlying defensive responding toward threat. As one of the first findings being confirmed by genome-wide association studies for the phenotype of panic disorder and agoraphobia, allelic variation in a gene coding for the glycine receptor beta subunit (GLRB) has recently been associated with increased neural fear network activation and enhanced acoustic startle reflexes. On the basis of two independent healthy control samples, we here aimed to further explore the functional significance of the GLRB genotype (rs7688285) by employing an intermediate phenotype approach. We focused on the phenotype of defensive system reactivity across the levels of brain function, structure, and physiology. Converging evidence across both samples was found for increased neurofunctional activation in the (anterior) insular cortex in GLRB risk allele carriers and altered fear conditioning as a function of genotype. The robustness of GLRB effects is demonstrated by consistent findings across different experimental fear conditioning paradigms and recording sites. Altogether, findings provide translational evidence for glycine neurotransmission as a modulator of the brain's evolutionary old dynamic defensive system and provide further support for a strong, biologically plausible candidate intermediate phenotype of defensive reactivity. As such, glycine-dependent neurotransmission may open up new avenues for mechanistic research on the etiopathogenesis of fear and anxiety disorders.


Serpent, suppressor of hairless and U-shaped are crucial regulators of hedgehog niche expression and prohemocyte maintenance during Drosophila larval hematopoiesis.
The lymph gland is a specialized organ for hematopoiesis, utilized during larval development in Drosophila. This tissue is composed of distinct cellular domains populated by blood cell progenitors (the medullary zone), niche cells that regulate the choice between progenitor quiescence and hemocyte differentiation [the posterior signaling center (PSC)], and mature blood cells of distinct lineages (the cortical zone). Cells of the PSC express the Hedgehog (Hh) signaling molecule, which instructs cells within the neighboring medullary zone to maintain a hematopoietic precursor state while preventing hemocyte differentiation. As a means to understand the regulatory mechanisms controlling Hh production, we characterized a PSC-active transcriptional enhancer that drives hh expression in supportive niche cells. Our findings indicate that a combination of positive and negative transcriptional inputs program the precise PSC expression of the instructive Hh signal. The GATA factor Serpent (Srp) is essential for hh activation in niche cells, whereas the Suppressor of Hairless [Su(H)] and U-shaped (Ush) transcriptional regulators prevent hh expression in blood cell progenitors and differentiated hemocytes. Furthermore, Srp function is required for the proper differentiation of niche cells. Phenotypic analyses also indicated that the normal activity of all three transcriptional regulators is essential for maintaining the progenitor population and preventing premature hemocyte differentiation. Together, these studies provide mechanistic insights into hh transcriptional regulation in hematopoietic progenitor niche cells, and demonstrate the requirement of the Srp, Su(H) and Ush proteins in the control of niche cell differentiation and blood cell precursor maintenance.


Selective Naked-Eye Detection of SARS-CoV-2 Mediated by N Gene Targeted Antisense Oligonucleotide Capped Plasmonic Nanoparticles.
The current outbreak of the pandemic coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) demands its rapid, convenient, and large-scale diagnosis to downregulate its spread within as well as across the communities. But the reliability, reproducibility, and selectivity of majority of such diagnostic tests fail when they are tested either to a viral load at its early representation or to a viral gene mutated during its current spread. In this regard, a selective "naked-eye" detection of SARS-CoV-2 is highly desirable, which can be tested without accessing any advanced instrumental techniques. We herein report the development of a colorimetric assay based on gold nanoparticles (AuNPs), when capped with suitably designed thiol-modified antisense oligonucleotides (ASOs) specific for N-gene (nucleocapsid phosphoprotein) of SARS-CoV-2, could be used for diagnosing positive COVID-19 cases within 10 min from the isolated RNA samples. The thiol-modified ASO-capped AuNPs agglomerate selectively in the presence of its target RNA sequence of SARS-CoV-2 and demonstrate a change in its surface plasmon resonance. Further, the addition of RNaseH cleaves the RNA strand from the RNA-DNA hybrid leading to a visually detectable precipitate from the solution mediated by the additional agglomeration among the AuNPs. The selectivity of the assay has been monitored in the presence of MERS-CoV viral RNA with a limit of detection of 0.18 ng/μL of RNA having SARS-CoV-2 viral load. Thus, the current study reports a selective and visual "naked-eye" detection of COVID-19 causative virus, SARS-CoV-2, without the requirement of any sophisticated instrumental techniques.


Innate lymphoid cells regulate CD4+ T-cell responses to intestinal commensal bacteria.
Innate lymphoid cells (ILCs) are a recently characterized family of immune cells that have critical roles in cytokine-mediated regulation of intestinal epithelial cell barrier integrity. Alterations in ILC responses are associated with multiple chronic human diseases, including inflammatory bowel disease, implicating a role for ILCs in disease pathogenesis. Owing to an inability to target ILCs selectively, experimental studies assessing ILC function have predominantly used mice lacking adaptive immune cells. However, in lymphocyte-sufficient hosts ILCs are vastly outnumbered by CD4(+) T cells, which express similar profiles of effector cytokines. Therefore, the function of ILCs in the presence of adaptive immunity and their potential to influence adaptive immune cell responses remain unknown. To test this, we used genetic or antibody-mediated depletion strategies to target murine ILCs in the presence of an adaptive immune system. We show that loss of retinoic-acid-receptor-related orphan receptor-γt-positive (RORγt(+)) ILCs was associated with dysregulated adaptive immune cell responses against commensal bacteria and low-grade systemic inflammation. Remarkably, ILC-mediated regulation of adaptive immune cells occurred independently of interleukin (IL)-17A, IL-22 or IL-23. Genome-wide transcriptional profiling and functional analyses revealed that RORγt(+) ILCs express major histocompatibility complex class II (MHCII) and can process and present antigen. However, rather than inducing T-cell proliferation, ILCs acted to limit commensal bacteria-specific CD4(+) T-cell responses. Consistent with this, selective deletion of MHCII in murine RORγt(+) ILCs resulted in dysregulated commensal bacteria-dependent CD4(+) T-cell responses that promoted spontaneous intestinal inflammation. These data identify that ILCs maintain intestinal homeostasis through MHCII-dependent interactions with CD4(+) T cells that limit pathological adaptive immune cell responses to commensal bacteria.


Efficacy, Safety, and Durability of Repeated Ketamine Infusions for Comorbid Posttraumatic Stress Disorder and Treatment-Resistant Depression.
The present study examined the efficacy, safety, and durability of repeated ketamine infusions for the treatment of comorbid posttraumatic stress disorder (PTSD) and treatment-resistant depression (TRD) in a sample of veterans. Individuals with comorbid DSM-5-defined PTSD and DSM-IV-defined major depressive disorder (N = 15) received 6 intravenous ketamine infusions (0.5 mg/kg) on a Monday-Wednesday-Friday schedule over a 12-day period from May 2015 to June 2016. Data from outcome measures were collected before and 24 hours after each infusion and weekly for 8 weeks following the final infusion. Continuous measures of symptom change were significant for both disorders and were associated with large effect sizes (mean decrease in PTSD Checklist for DSM-5 score = 33.3 points [95% CI, 23.0-43.5 points], P < .0005, sample size-adjusted Cohen d [d'] = 2.17; mean decrease in Montgomery-Asberg Depression Rating Scale score = 26.6 points [95% CI, 23.0-30.2 points], P < .0005, d' = 4.64). The remission rate for PTSD was 80.0%, and the response rate for TRD was 93.3%. Participants in remission from PTSD after the infusion series (n = 12) had a median time to relapse of 41 days. Similarly, participants whose depression symptoms responded to the infusion series (n = 14) had a median time to relapse of 20 days. Repeated ketamine infusions were associated with transient increases in dissociative symptoms. No participant reported worsening of PTSD symptoms over the study duration. This study, the first open-label study of repeated ketamine infusions in a comorbid population, found rapid and sustained improvement in PTSD and depression symptoms. This report suggests that repeated ketamine treatments are safe and may represent an efficacious treatment for individuals with comorbid PTSD and TRD. ClinicalTrials.gov identifier: NCT02577250.


Pharmaceutical services in hospital-based home health-care agencies--1982.
A survey of Medicare-certified hospital-based home health-care agencies was conducted in May 1982 to determine the type and extent of pharmaceutical services provided. A 12-page questionnaire was mailed to 243 directors of U.S. agencies that were identified from a 1976 directory. The questions elicited information about the characteristics of the director and agency, personnel, reimbursement, and scope of services. The overall response rate was 73.7%. Ninety-five percent of the agency directors were nurses. The median patient census of the agencies was 110. All agencies reported offering visiting-nurse services, 92% provided home health-aide services, and 46% offered home hospice services. Ninety-nine percent and 91% reported receiving reimbursement from Medicare and Medicaid, respectively. One fourth of the agencies provided home services that traditionally involve hospital pharmacists, including intravenous therapy (29%), home chemotherapy (18%), and total parenteral nutrition (18%). Although 85% of the agency directors reported using the services of a pharmacist, only 4% actually employed a pharmacist on their staff. The directors viewed educational programs, drug regimen review, and drug information services as the most important functions of pharmacists in home health-care agencies. Although these agencies provided an array of pharmaceutical services in 1982, very few pharmacists were actually employed. Additional studies are needed to re-evaluate the current status of pharmacy involvement in home health care.


Teratogenic Effects of Third-Generation Antiepileptic Drug, Pregabalin: An In vivo Study.
In the recent past, many third-generation antiepileptic drugs (AEDs) including Pregabalin (PGB) were launched for the treatment of diverse forms of epilepsy with better efficacy and safety profile than first-and-second-generation AEDs, but their teratogenic safety has not been established so far. The present study has been undertaken to evaluate the reproductive and teratogenic potential (external and skeletal) of a novel and third generation AED, PGB in pregnant albino rats. In this study, pregnant subjects were exposed to clinically relevant doses (41, 82 and 123 mg) of PGB from gestation days 6-20, and sacrificed on GD-21, and their fetuses were collected and examined to identify the birth defects and skeletal anomalies. This study revealed that prenatal exposure to PGB induced dose-dependent substantial fetal resorptions, litter size, fetal length and weight; and variety of minor external and internal malformations in fetuses predominant with limbs, tail, eyes, abdomen including hemorrhages, and poor skeletal ossification. Thus, PGB was found to be teratogenic in rats at equivalent therapeutic doses, hence precaution should be taken before prescribing PGB to pregnant women with epilepsy.


Cannabidiol-from Plant to Human Body: A Promising Bioactive Molecule with Multi-Target Effects in Cancer.
Cannabis sativa L. is a plant long used for its textile fibers, seed oil, and oleoresin with medicinal and psychoactive properties. It is the main source of phytocannabinoids, with over 100 compounds detected so far. In recent years, a lot of attention has been given to the main phytochemicals present in Cannabis sativa L., namely, cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC). Compared to THC, CBD has non-psychoactive effects, an advantage for clinical applications of anti-tumor benefits. The review is designed to provide an update regarding the multi-target effects of CBD in different types of cancer. The main focus is on the latest in vitro and in vivo studies that present data regarding the anti-proliferative, pro-apoptotic, cytotoxic, anti-invasive, anti-antiangiogenic, anti-inflammatory, and immunomodulatory properties of CBD together with their mechanisms of action. The latest clinical evidence of the anticancer effects of CBD is also outlined. Moreover, the main aspects of the pharmacological and toxicological profiles are given.


Localization of fibronectin matrix assembly sites on fibroblasts and endothelial cells.
Polymerization of soluble fibronectin into extracellular matrix fibers occurs through the interaction between the amino terminus of fibronectin contained within a 70 kDa fragment and 'matrix assembly sites' on the cell surface. The present studies were performed to localize the 'matrix assembly sites' (defined by 70 kDa binding sites) on newly adherent cells and on cells containing preformed fibronectin matrix. Matrix nucleation sites on newly spread cells were visualized using Texas Red conjugated 70 kDa fragment and were found to colocalize with vinculin and substrate fibronectin fibrils. Cells plated onto vitronectin coated coverslips did not exhibit any 70 kDa binding sites although these cells were well-spread with fully developed focal adhesions. Time course studies indicated that 70 kDa binding sites could be detected on newly adherent cells within 30-40 minutes following cell plating onto fibronectin coated coverslips, prior to the reorganization of substrate fibronectin into fibrils. Similarly, exogenous fibronectin conjugated with Texas Red was also colocalized with vinculin when added to newly adherent cells. The disruption of actin filaments with cytochalasin D both prevented the expression of 70 kDa binding sites and also resulted in the loss of established 70 kDa binding sites on newly spread cells. After 3 days in culture, cells organized an extensive fibronectin matrix and 70 kDa was colocalized with two distinct types of matrix fibronectin fibers: fine linear cell-associated fibers which co-stained with the beta1 integrin and coarse extracellular fibers which did not stain for the beta1 integrin. There was also a third type of fibronectin fiber which was organized into a meshwork structure. There was no localization of either beta1 or 70 kDa to these structures. Treatment of 3-day cells with cytochalasin D resulted in the disruption of cell-matrix fibers and cell-associated 70 kDa binding sites. In contrast, the coarse extracellular matrix fibers as well as the meshwork fibers were unaffected by cytochalasin. In the presence of cytochalasin D, 70 kDa bound to sites which colocalized with the coarse extracellular matrix fibers. These data suggest that de novo assembly of fibronectin matrix occurs at sites of focal adhesion and as fibronectin polymerization proceeds, matrix nucleation sites colocalize along cell associated fibronectin fibers. At later times 70 kDa is localized to a subset of more mature fibronectin-containing fibers. These results suggest that there are at least three morphologically distinct 70 kDa binding sites on adherent cells: one which colocalizes with beta1 to focal adhesions, a second which colocalizes with beta1 and fibronectin in matrix contacts, and a third which localizes to extracellular matrix fibers.


Fourier-transform method for the design of wideband antireflection coatings.
An iterative correction process, recently incorporated into the National Research Council of Canada Fourier-transform thin-film synthesis program, is applied to the design of wideband antireflection coatings. This type of problem is different from those solved in the past by this method. It cannot be handled in a practical way without a correction process. We consider in detail the effects-critical for this application-of constraints on the refractive indices and overall thicknesses of the solutions. Our graded-index and multilayer designs have a remarkable resemblance in performance and refractive-index structure to results obtained by more conventional techniques. The Fourier-transform method is of interest because of its speed and versatility.


WSB-1 regulates the metastatic potential of hormone receptor negative breast cancer.
Metastatic spread is responsible for the majority of cancer-associated deaths. The tumour microenvironment, including hypoxia, is a major driver of metastasis. The aim of this study was to investigate the role of the E3 ligase WSB-1 in breast cancer biology in the context of the hypoxic tumour microenvironment, particularly regarding metastatic spread. In this study, WSB-1 expression was evaluated in breast cancer cell lines and patient samples. In silico analyses were used to determine the impact of WSB-1 expression on distant metastasis-free survival (DMFS) in patients, and correlation between WSB1 expression and hypoxia gene expression signatures. The role of WSB-1 on metastasis promotion was evaluated in vitro and in vivo. High WSB1 expression was associated with decreased DMFS in ER-breast cancer and PR-breast cancer patients. Surprisingly, WSB1 expression was not positively correlated with known hypoxic gene expression signatures in patient samples. Our study is the first to show that WSB-1 knockdown led to decreased metastatic potential in breast cancer hormone receptor-negative models in vitro and in vivo. WSB-1 knockdown was associated with decreased metalloproteinase (MMP) activity, vascular endothelial growth factor (VEGF) secretion, and angiogenic potential. Our data suggests that WSB-1 may be an important regulator of aggressive metastatic disease in hormone receptor-negative breast cancer. WSB-1 could therefore represent a novel regulator and therapeutic target for secondary breast cancer in these patients.


Chelation-directed C-H activation/C-C bond forming reactions catalyzed by Pd(ii) nanoparticles supported on multiwalled carbon nanotubes.
Chelation-directed C-H activation/C-C bond forming reactions utilizing homogeneous palladium(ii) and the Pd(ii)/Pd(iv) catalytic cycle have been previously reported. Here we report the first use of a solid-supported Pd(ii) catalyst [Pd(ii) nanoparticles on multiwalled carbon nanotubes, Pd(ii)/MWCNT] to carry out C-H activation/C-C bond forming reactions. The results presented demonstrate that the solid-supported Pd(ii)/MWCNT catalyst can effectively catalyze these arylation reactions using the Pd(ii)/Pd(iv) catalytic cycle. We also show that the solid-supported catalyst is recyclable, has turnover frequencies up to 2.9-fold higher than the homogeneous catalyst, and results in low levels of residual palladium contamination in the products.


Why weight? Modelling sample and observational level variability improves power in RNA-seq analyses.
Variations in sample quality are frequently encountered in small RNA-sequencing experiments, and pose a major challenge in a differential expression analysis. Removal of high variation samples reduces noise, but at a cost of reducing power, thus limiting our ability to detect biologically meaningful changes. Similarly, retaining these samples in the analysis may not reveal any statistically significant changes due to the higher noise level. A compromise is to use all available data, but to down-weight the observations from more variable samples. We describe a statistical approach that facilitates this by modelling heterogeneity at both the sample and observational levels as part of the differential expression analysis. At the sample level this is achieved by fitting a log-linear variance model that includes common sample-specific or group-specific parameters that are shared between genes. The estimated sample variance factors are then converted to weights and combined with observational level weights obtained from the mean-variance relationship of the log-counts-per-million using 'voom'. A comprehensive analysis involving both simulations and experimental RNA-sequencing data demonstrates that this strategy leads to a universally more powerful analysis and fewer false discoveries when compared to conventional approaches. This methodology has wide application and is implemented in the open-source 'limma' package.


Synergistic effect on cardiac energetics by targeting the creatine kinase system: in vivo application of high-resolution 31P-CMRS in the mouse.
Phosphorus cardiovascular magnetic resonance spectroscopy (31P-CMRS) has emerged as an important tool for the preclinical assessment of myocardial energetics in vivo. However, the high rate and diminutive size of the mouse heart is a challenge, resulting in low resolution and poor signal-to-noise. Here we describe a refined high-resolution 31P-CMRS technique and apply it to a novel double transgenic mouse (dTg) with elevated myocardial creatine and creatine kinase (CK) activity. We hypothesised a synergistic effect to augment energetic status, evidenced by an increase in the ratio of phosphocreatine-to-adenosine-triphosphate (PCr/ATP). Single transgenic Creatine Transporter overexpressing (CrT-OE, n = 7) and dTg mice (CrT-OE and CK, n = 6) mice were anaesthetised with isoflurane to acquire 31P-CMRS measurements of the left ventricle (LV) utilising a two-dimensional (2D), threefold under-sampled density-weighted chemical shift imaging (2D-CSI) sequence, which provided high-resolution data with nominal voxel size of 8.5 µl within 70 min. (1H-) cine-CMR data for cardiac function assessment were obtained in the same imaging session. Under a separate examination, mice received invasive haemodynamic assessment, after which tissue was collected for biochemical analysis. Myocardial creatine levels were elevated in all mouse hearts, but only dTg exhibited significantly elevated CK activity, resulting in a 51% higher PCr/ATP ratio in heart (3.01 ± 0.96 vs. 2.04 ± 0.57-mean ± SD; dTg vs. CrT-OE), that was absent from adjacent skeletal muscle. No significant differences were observed for any parameters of LV structure and function, confirming that augmentation of CK activity does not have unforeseen consequences for the heart. We have developed an improved 31P-CMRS methodology for the in vivo assessment of energetics in the murine heart which enabled high-resolution imaging within acceptable scan times. Mice over-expressing both creatine and CK in the heart exhibited a synergistic elevation in PCr/ATP that can now be tested for therapeutic potential in models of chronic heart failure.


Stereochemistry of post-transition metal oxides: revision of the classical lone pair model.
The chemistry of post transition metals is dominated by the group oxidation state N and a lower N-2 oxidation state, which is associated with occupation of a metal s(2) lone pair, as found in compounds of Tl(I), Pb(II) and Bi(III). The preference of these cations for non-centrosymmetric coordination environments has previously been rationalised in terms of direct hybridisation of metal s and p valence orbitals, thus lowering the internal electronic energy of the N-2 ion. This explanation in terms of an on-site second-order Jahn-Teller effect remains the contemporary textbook explanation. In this tutorial review, we review recent progress in this area, based on quantum chemical calculations and X-ray spectroscopic measurements. This recent work has led to a revised model, which highlights the important role of covalent interaction with oxygen in mediating lone pair formation for metal oxides. The role of the anion p atomic orbital in chemical bonding is key to explaining why chalcogenides display a weaker preference for structural distortions in comparison to oxides and halides. The underlying chemical interactions are responsible for the unique physicochemical properties of oxides containing lone pairs and, in particular, to their application as photocatalysts (BiVO(4)), ferroelectrics (PbTiO(3)), multi-ferroics (BiFeO(3)) and p-type semiconductors (SnO). The exploration of lone pair systems remains a viable a venue for the design of functional multi-component oxide compounds.


Network analysis of depressive symptoms in Hong Kong residents during the COVID-19 pandemic.
In network theory depression is conceptualized as a complex network of individual symptoms that influence each other, and central symptoms in the network have the greatest impact on other symptoms. Clinical features of depression are largely determined by sociocultural context. No previous study examined the network structure of depressive symptoms in Hong Kong residents. The aim of this study was to characterize the depressive symptom network structure in a community adult sample in Hong Kong during the COVID-19 pandemic. A total of 11,072 participants were recruited between 24 March and 20 April 2020. Depressive symptoms were measured using the Patient Health Questionnaire-9. The network structure of depressive symptoms was characterized, and indices of "strength", "betweenness", and "closeness" were used to identify symptoms central to the network. Network stability was examined using a case-dropping bootstrap procedure. Guilt, Sad Mood, and Energy symptoms had the highest centrality values. In contrast, Concentration, Suicide, and Sleep had lower centrality values. There were no significant differences in network global strength (p = 0.259), distribution of edge weights (p = 0.73) and individual edge weights (all p values > 0.05 after Holm-Bonferroni corrections) between males and females. Guilt, Sad Mood, and Energy symptoms were central in the depressive symptom network. These central symptoms may be targets for focused treatments and future psychological and neurobiological research to gain novel insight into depression.


Increased efficiency in identifying mixed pollen samples by meta-barcoding with a dual-indexing approach.
Meta-barcoding of mixed pollen samples constitutes a suitable alternative to conventional pollen identification via light microscopy. Current approaches however have limitations in practicability due to low sample throughput and/or inefficient processing methods, e.g. separate steps for amplification and sample indexing. We thus developed a new primer-adapter design for high throughput sequencing with the Illumina technology that remedies these issues. It uses a dual-indexing strategy, where sample-specific combinations of forward and reverse identifiers attached to the barcode marker allow high sample throughput with a single sequencing run. It does not require further adapter ligation steps after amplification. We applied this protocol to 384 pollen samples collected by solitary bees and sequenced all samples together on a single Illumina MiSeq v2 flow cell. According to rarefaction curves, 2,000-3,000 high quality reads per sample were sufficient to assess the complete diversity of 95% of the samples. We were able to detect 650 different plant taxa in total, of which 95% were classified at the species level. Together with the laboratory protocol, we also present an update of the reference database used by the classifier software, which increases the total number of covered global plant species included in the database from 37,403 to 72,325 (93% increase). This study thus offers improvements for the laboratory and bioinformatical workflow to existing approaches regarding data quantity and quality as well as processing effort and cost-effectiveness. Although only tested for pollen samples, it is furthermore applicable to other research questions requiring plant identification in mixed and challenging samples.


The effects of phthalate and nonylphenol exposure on body size and secondary sexual characteristics during puberty.
Some phthalic acid esters (PAEs) and nonylphenol (NP) are endocrine-disrupting chemicals (EDCs) that are widely used in consumer products. Consequently, the general population is exposed simultaneously to both groups of chemicals. To investigate the single- and co-exposure effects of PAEs (DMP, DEP, DnBP, DiBP, BBzP, and DEHP) and NP on obesity and pubertal maturity to compare the body sizes of general adolescents with the complainants of the phthalate-tainted foods scandal that occurred in Taiwan. This study included 270 general adolescents aged 6.5-15.0 years and 38 complainants aged 6.5-8.5 years. Nine metabolites of the five PAEs and of NP were measured in urine. We used a questionnaire to evaluate pubertal maturity, measured anthropometric indices (APs) to assess body size, and collected urine samples to measure the two groups of chemicals. We found that urinary PAE metabolite concentrations (specifically, metabolites of DEP, DnBP, DiBP, and DEHP) were positively associated with the APs for abdominal obesity (including skinfold thickness, waist circumference, waist-to-height ratio, and waist-to-hip) and indicated a dose-response relationship. Mono-methyl phthalate (MMP) exposure was inversely associated with pubarche among boys. The daily intake of DEHP in general adolescents exceeded the reference doses (RfD-20 μg/kgbw/day) and tolerable daily intake (TDI-50 μg/kgbw/day) by 3.4% and 0.4%, respectively. No associations were observed between NP exposure or co-exposure and the APs or pubertal maturity. No significant differences were observed between general adolescents and the complainants with regard to weight, height, or BMI. The study suggests that PAE (specifically, DEP, DnBP, DiBP, and DEHP) exposure is associated with abdominal obesity in adolescents and that the APs for abdominal obesity are more sensitive than BMI for measuring obesity among adolescents. We suggest that the RfD and TDI for PAEs should be revised to provide sufficient protection.


Health resource utilization associated with skeletal-related events: results from a retrospective European study.
Bone complications, also known as skeletal-related events (SREs), are common in patients with bone metastases secondary to advanced cancers. To provide a detailed estimate of the health resource utilization (HRU) burden associated with SREs across eight European countries. Eligible patients from centers in Austria, the Czech Republic, Finland, Greece, Poland, Portugal, Sweden, and Switzerland with bone metastases or lesions secondary to breast cancer, prostate, or lung cancer or multiple myeloma who had experienced at least one SRE (defined as radiation to bone, long-bone pathologic fracture, other bone pathologic fracture, surgery to bone or spinal cord compression) were entered into this study. HRU data were extracted retrospectively from the patients' charts from 3.5 months before the index SRE until 3 months after the index SRE (defined as an SRE preceded by an SRE-free period of at least 6.5 months). Overall, the mean number of inpatient stays per SRE increased from baseline by approximately 0.5-1.5 stays, with increases in the total duration of inpatient stays of approximately 6-37 days per event. All SREs were associated with substantial increases from baseline in the frequency of procedures and the number of outpatient and day-care visits. SREs are associated with substantial HRU owing to considerable increases in the number and duration of inpatient stays, and in the number of procedures, outpatient visits, and day-care visits. These data collectively provide a valuable summary of the real-world SRE burden on European healthcare systems.