Which steps are essential for the formation of functional legume nodules?
Nodulation is reviewed in terms of the phenotypes proposed by Vincent (1980). Individual legumes may be infectible by one or more of the three bacterial genera (collectively known as rhizobia) Rhizobium, Bradyrhizobium, or Azorhizobium. The type of infection process by which rhizobia gain entry is largely governed by the host genotype. In addition to the widely studied root-hair pathway, infections may be associated with lateral root emergence or occur between root epidermal cells. The exact chemical and physical nature of the root hair/epidermal cell wall is likely to be a critical factor in determining whether infections can proceed. In addition to differing with species, wall composition may be influenced by soil chemical (e.g. Ca2+ ) and biotic factors (e.g. bacteria). Rhizobial features essential for infection include particular surface polysaccharides and the induction of nodulation genes by plant root exudates. Neither of these is likely to be a major barrier to the extension of nodulation to new hosts. Dissemination of rhizobia within developing nodules may be intercellular, via infection threads or by division of a small number of infected cells. All functional symbioses eventually have 'intracellular' bacteria, in the sense that rhizobia are geographically located within the boundary of the host cell walls. However, they remain extracellular in the sense that they are always confined by a membrane which is largely of host cell origin. In some genera they are also surrounded by infection thread walls, probably modified forms of 'invasive' infection thread walls, which allow differentiation of rhizobia into the nitrogen-fixing form. Thus, natural, functional, symbioses may (a) never involve a stage in which bacteria are confined within tubular infection threads or (b) never release bacteria from infection threads. These features are determined by host genotype. The one feature of legume nodules so far found never to vary is the stem-like character of a peripheral vascular system. This contrasts with the central vascular system of actinorhizas and the rhizobial-induced nodules on the Ulmaceous genus Parasponia. Although of great intrinsic interest, this character is unlikely to present an insurmountable barrier to the extension of nodulation to new species. Other features, such as the ability to produce haemoglobin are now known to the in the genetic makeup of many higher plants. The discovery of the wide range of nodule structures occurring in nature, together with work on mutant rhizobia which may bypass critical stages in the nodulation process, suggest various ways in which the extension of nodulation to non-nodulated legumes and to other (initially at least, dicotyledonous) plants may be engineered. CONTENTS Summary 129 I. Introduction 130 II. The symbionts 130 III. Stages in nodulation 132 IV. Stems and nodules 143 V. Prospects for finding/making new symbioses 144 VI. Conclusions 145 Acknowledgements 147 References 147.


Maternal and infant characteristics by mode of vitamin K prophylaxis administration.
The aim of this study was to compare maternal and infant characteristics by mode of VK administration. De-identified computerised birth files of all babies born in New South Wales (NSW), Australia between January 2007 and December 2009 (when VK prophylaxis was measured) were included in the present study. The outcome variable, mode of VK prophylaxis, was recorded by checkbox as oral, IM injection, none or not stated. We analysed population-based birth data from 2007 to 2009 in NSW, Australia and found that IM injection was the most prevalent mode of administration (96.3%, n = 263, 555), followed by oral (2.6%, n = 7023) and none (1.2%, n = 3136). Compared to neonates receiving IM VK, those with oral or none were more likely to have vaginal births without medical interventions at birth centres or planned home births and were less likely to receive hepatitis B vaccination. Among neonates administered oral VK, a larger proportion were preterm births and breastfed at discharge compared to neonates administered VK as an IM injection. Neonates with no VK recorded were more likely to be admitted to neonatal intensive care, but may have received VK later in the birth admission. A small proportion of the Australian neonates may be at risk of inadequate protection from VKBD due to parental concerns about the safety of IM injection of VK to neonates.


Coordination versus coupling of dicyanamide in molybdenum and manganese pyrazole complexes.
The reactions of cis-[MoCl(η(3)-methallyl)(CO)(2)(NCMe)(2)] (methallyl = CH(2)C(CH(3))CH(2)) with Na(NCNCN) and pz*H (pzH, pyrazole, or dmpzH, 3,5-dimethylpyrazole) lead to cis-[Mo(η(3)-methallyl)(CO)(2)(pz*H)(μ-NCNCN-κ(2)N,N)](2) (pzH, 1a; dmpzH, 1b), where dicyanamide is coordinated as bridging ligand. Similar reactions with fac-[MnBr(CO)(3)(NCMe)(2)] lead to the pyrazolylamidino complexes fac-[Mn(pz*H)(CO)(3)(NH═C(pz*)NCN-κ(2)N,N)] (pzH, 2a; dmpzH, 2b), resulting from the coupling of pyrazol with one of the CN bonds of dicyanamide. The second CN bond of dicyanamide in 2a undergoes a second coupling with pyrazole after addition of 1 equiv of fac-[MnBr(CO)(3)(pzH)(2)], yielding the dinuclear doubly coupled complex [{fac-Mn(pzH)(CO)(3)}(2)(μ-NH═C(pz)NC(pz)=NH-κ(4)N,N,N,N)]Br (3). The crystal structure of 3 reveals the presence of two isomers, cis or trans, depending on whether the terminal pyrazoles are coordinated at the same or at different sides of the approximate plane defined by the bridging bis-amidine ligand. Only the cis isomer is detected in the crystal structure of the perchlorate salt of the same bimetallic cation (4), obtained by metathesis with AgClO(4). All the N-bound hydrogen atoms of the cations in 3 or 4 are involved in hydrogen bonds. Some of the C-N bonds of the pyrazolylamidino ligand have a character intermediate between single and double, and theoretical studies were carried out on 2a and 3 to confirm its electronic origin and discard packing effects. Calculations also show the essential role of bromide in the planarity of the tetradentate ligand in the bimetallic complex 3.


Universal vaccination of children against hepatitis A in Chile: a cost-effectiveness study.
To evaluate the healthcare and economic impact of routine hepatitis A vaccination of toddlers in Chile. We used a dynamic model of hepatitis A infection to evaluate the impact of a two-dose vaccination program, administered at ages 12 and 18 months. The model incorporated the changing epidemiology of hepatitis A in Chile and the development of vaccine-induced herd immunity. Our analysis was conducted from the public payer perspective, and an estimation of the societal perspective was performed. Costs are expressed in 2005 U.S. dollars. Vaccination of toddlers rapidly reduced the healthcare burden of hepatitis A. In the base case (95% vaccination coverage, 100-year time horizon, 1% annual decrease in force of infection), the average number of infections fell by 76.6% annually, and associated deaths fell by 59.7%. Even at 50% coverage, the program reduced infection rates substantially. Routine vaccination of toddlers had economic as well as health benefits, saving $4,984 per life-year gained (base case scenario). The program became cost saving after 6 years, and its overall cost-effectiveness per life-year gained was largely unaffected by changes in disease-related costs, herd immunity, coverage rate, and annual decrease in force of infection. Routine vaccination of toddlers will reduce the rates of symptomatic hepatitis A and associated mortality. The two-dose schedule evaluated here will be less expensive than disease-related costs in the absence of vaccination from the sixth year of its implementation. These findings support the establishment of a routine vaccination program for toddlers in Chile.


c-mip impairs podocyte proximal signaling and induces heavy proteinuria.
Idiopathic nephrotic syndrome comprises several podocyte diseases of unknown origin that affect the glomerular podocyte, which controls the permeability of the filtration barrier in the kidney to proteins. It is characterized by the daily loss of more than 3 g of protein in urine and the lack of inflammatory lesions or cell infiltration. We found that the abundance of c-mip (c-maf inducing protein) was increased in the podocytes of patients with various acquired idiopathic nephrotic syndromes in which the podocyte is the main target of injury. Mice engineered to have excessive c-mip in podocytes developed proteinuria without morphological alterations, inflammatory lesions, or cell infiltration. Excessive c-mip blocked podocyte signaling by preventing the interaction of the slit diaphragm transmembrane protein nephrin with the tyrosine kinase Fyn, thereby decreasing phosphorylation of nephrin in vitro and in vivo. Moreover, c-mip inhibited interactions between Fyn and the cytoskeletal regulator N-WASP (neural Wiskott-Aldrich syndrome protein) and between the adaptor protein Nck and nephrin, potentially accounting for cytoskeletal disorganization and the effacement of foot processes seen in idiopathic nephrotic syndromes. The intravenous injection of small interfering RNA targeting c-mip prevented lipopolysaccharide-induced proteinuria in mice. Together, these results identify c-mip as a key component in the molecular pathogenesis of acquired podocyte diseases.


Risk of Psychiatric Disorders Among Individuals With the 22q11.2 Deletion or Duplication: A Danish Nationwide, Register-Based Study.
Microdeletions and duplications have been described at the 22q11.2 locus. However, little is known about the clinical and epidemiologic consequences at the population level. To identify indicators of deletions or duplications at the 22q11.2 locus and estimate the incidence rate ratios (IRRs) and absolute risk for psychiatric disorders in clinically identified individuals with 22q11.2 deletion or duplication. A Danish nationwide register study including all individuals recorded in the Danish Cytogenetic Central Register with a 22q11.2 deletion or duplication was performed. A total of 3 768 943 individuals born in Denmark from 1955 to 2012 were followed up during the study period (total follow-up, 57.1 million person-years). Indicators of 22q11.2 deletion or duplication and cumulative incidences were estimated using a nested case-control design that included individuals from the population-based cohort. Survival analysis was used to compare risk of disease in individuals with and without the 22q11.2 deletion or duplication. The study was conducted from May 7, 2015, to August 14, 2016. The 22q11.2 deletion or duplication. Indicators for carrying a 22q11.2 deletion or duplication, IRR, and cumulative incidences for psychiatric diagnoses (International Statistical Classification of Diseases and Related Health Problems, 10th Revision, codes F00-F99), including schizophrenia-spectrum disorders, mood disorders, neurotic stress-related and somatoform disorders, and a range of developmental and childhood disorders. Among the 3 768 943 participants, 244 (124 [50.8%] male) and 58 (29 [50.0%] male) individuals were clinically identified with a 22q11.2 deletion or duplication, respectively. Mean (SD) age at diagnosis of any psychiatric disorder was 12.5 (8.3) years for individuals with deletions and 6.1 (0.9) years for duplication carriers. A parental diagnosis of schizophrenia-but not of other psychiatric diagnoses-was associated with a 22q11.2 deletion, and parental psychiatric diagnoses other than schizophrenia were associated with duplication carrier status. Both the 22q11.2 deletion (IRR, 4.24; 95% CI, 3.07-5.67) and duplication (IRR, 4.99; 95% CI, 1.79-10.72) was associated with increased risk of any psychiatric disorders. Furthermore, a highly increased risk of intellectual disability was found for the deletion (IRR, 34.08; 95% CI, 22.39-49.27) and duplication (IRR, 33.86; 95% CI, 8.42-87.87). Furthermore, individuals with the 22q11.2 deletion had an increased risk of several psychiatric disorders under study, for example, pervasive developmental disorders (IRR, 9.45; 95% CI, 5.64-14.69) and childhood autism (IRR, 8.94; 95% CI, 3.21-19.23). Individuals with the 22q11.2 deletion or duplication have a significantly increased risk of developing psychiatric disorders. Survival analysis of persons carrying either the 22q11.2 deletion or duplication provides estimates of direct clinical relevance useful to assist clinical ascertainment, genetic counseling, guidance of symptomatic monitoring, and early clinical intervention.


Ultrastructural features of fetal erythroid precursors infected with parvovirus B19 in vitro: evidence of cell death by apoptosis.
Human parvovirus B19 cannot be cultured in standard cell lines and relatively little is known about the intracellular life-cycle of the virus. In this study, ultrastructural features of B19 infection were examined using haemopoietic cell suspension cultures derived from human fetal liver. Erythroblasts from infected cultures frequently contained crystalline arrays of both full and empty virus-like particles. The number and size of these arrays increased with the duration of culture, and their location changed from exclusively nuclear at 24 h post-infection to both nuclear and cytoplasmic at 3 days post-infection. Arrays were occasionally found in cytoplasmic protuberances which appeared to be pinching off from the cell. The location of the arrays corresponded to the distribution of viral capsid protein determined by immunolabelling at the light microscope level. Cells containing viral crystalline arrays also exhibited nucleolar degeneration, extreme margination of the nuclear heterochromatin, and cytoplasmic vacuolation. These features are typical of cells undergoing individual programmed cell death or 'apoptosis'. The triggering of apoptosis in erythroid precursors by parvovirus B19 may help to explain the apparent lack of a strong inflammatory response to fetal B19 infection and may have implications for understanding the mechanisms of viral spread throughout the host.


Bidirectional-Compounding Effects of Rumination and Negative Emotion in Predicting Impulsive Behavior: Implications for Emotional Cascades.
Influenced by chaos theory, the emotional cascade model proposes that rumination and negative emotion may promote each other in a self-amplifying cycle that increases over time. Accordingly, exponential-compounding effects may better describe the relationship between rumination and negative emotion when they occur in impulsive persons, and predict impulsive behavior. Forty-seven community and undergraduate participants who reported frequent engagement in impulsive behaviors monitored their ruminative thoughts and negative emotion multiple times daily for two weeks using digital recording devices. Hypotheses were tested using cross-lagged mixed model analyses. Findings indicated that rumination predicted subsequent elevations in rumination that lasted over extended periods of time. Rumination and negative emotion predicted increased levels of each other at subsequent assessments, and exponential functions for these associations were supported. Results also supported a synergistic effect between rumination and negative emotion, predicting larger elevations in subsequent rumination and negative emotion than when one variable alone was elevated. Finally, there were synergistic effects of rumination and negative emotion in predicting number of impulsive behaviors subsequently reported. These findings are consistent with the emotional cascade model in suggesting that momentary rumination and negative emotion progressively propagate and magnify each other over time in impulsive people, promoting impulsive behavior.


Exploring the shape deformation of biomembrane tubes with theoretical analysis and computer simulation.
The shape deformation of membrane nanotubes is studied by a combination of theoretical analysis and molecular simulation. First we perform free energy analysis to demonstrate the effects of various factors on two ideal states for the pearling transition, and then we carry out dissipative particle dynamics simulations, through which various types of membrane tube deformation are found, including membrane pearling, buckling, and bulging. Different models for inducing tube deformation, including the osmotic pressure, area difference and spontaneous curvature models, are considered to investigate tubular instabilities. Combined with free energy analysis, our simulations show that the origin of the deformation of membrane tubes in different models can be classified into two categories: effective spontaneous curvature and membrane tension. We further demonstrate that for different models, a positive membrane tension is required for the pearling transition. Finally we show that different models can be coupled to effectively deform the membrane tube.


PLGA/hydrogel biopapers as a stackable substrate for printing HUVEC networks via BioLP.
Two major challenges in tissue engineering are mimicking the native cell-cell arrangements of tissues and maintaining viability of three-dimension (3D) tissues thicker than 300 µm. Cell printing and prevascularization of engineered tissues are promising approaches to meet these challenges. However, the printing technologies used in biofabrication must balance the competing parameters of resolution, speed, and volume, which limit the resolution of thicker 3D structures. We suggest that high-resolution conformal printing techniques can be used to print 2D patterns of vascular cells onto biopaper substrates which can then be stacked to form a thicker tissue construct. Towards this end we created 1 cm × 1 cm × 300 µm biopapers to be used as the transferable, stackable substrate for cell printing. 3.6% w/v poly-lactide-co-glycolide was dissolved in chloroform and poured into molds filled with NaCl crystals. The salt was removed with DI water and the scaffolds were dried and loaded with a Collagen Type I or Matrigel. SEM of the biopapers showed extensive porosity and gel loading throughout. Biological laser printing (BioLP) was used to deposit human umbilical vein endothelial cells (HUVEC) in a simple intersecting pattern to the surface of the biopapers. The cells differentiated and stretched to form networks preserving the printed pattern. In a separate experiment to demonstrate "stackability," individual biopapers were randomly seeded with HUVECs and cultured for 1 day. The mechanically stable and viable biopapers were then stacked and cultured for 4 days. Three-dimensional confocal microscopy showed cell infiltration and survival in the compound multilayer constructs. These results demonstrate the feasibility of stackable "biopapers" as a scaffold to build 3D vascularized tissues with a 2D cell-printing technique.


The influence of cannulation technique on blood flow to the brain in rats undergoing cardiopulmonary bypass: a cautionary "tail".
Recently, there has been an increase in the use of rat models of cardiopulmonary bypass (CPB) for research purposes. Much of this work has focused on cerebral injury associated with CPB. Many of these studies employ a peripheral cannulation approach, often utilizing the caudal artery and internal or external jugular vein. The aim of the present study was to establish whether there is any alteration in blood flow to the brain associated with the use of different cannulation routes. Twenty-four adult male Sprague Dawley rats were allocated to one of three study groups: Group 1- caudal artery return, Group 2 -open-chest aortic return, and Group 3- nonbypass control group. Colored microspheres were injected into all animals at four time points (postinduction, initiation of bypass, midbypass, and end bypass). After the termination of each experiment, the brains were excised, the tissue was digested, the microspheres were harvested, and the global blood flow to the brain was assessed using the reference flow method. There was a significant reduction in blood flow to the brain between both bypass groups and the control group. Additionally, cerebral blood flow was significantly lower in the caudal return group than in the aortic return group. There is a significant drop in blood flow to the brain associated with the initiation and continuation of CPB when compared to non-CPB controls. These results also confirm a considerable cerebral hypoperfusion associated with the peripheral cannulation technique, and suggest that peripheral bypass may exaggerate the influence CPB has on cerebral injury. This technique must therefore be employed with caution.


Copy number variation in obsessive-compulsive disorder and tourette syndrome: a cross-disorder study.
Obsessive-compulsive disorder (OCD) and Tourette syndrome (TS) are heritable neurodevelopmental disorders with a partially shared genetic etiology. This study represents the first genome-wide investigation of large (>500 kb), rare (<1%) copy number variants (CNVs) in OCD and the largest genome-wide CNV analysis in TS to date. The primary analyses used a cross-disorder design for 2,699 case patients (1,613 ascertained for OCD, 1,086 ascertained for TS) and 1,789 controls. Parental data facilitated a de novo analysis in 348 OCD trios. Although no global CNV burden was detected in the cross-disorder analysis or in secondary, disease-specific analyses, there was a 3.3-fold increased burden of large deletions previously associated with other neurodevelopmental disorders (p = .09). Half of these neurodevelopmental deletions were located in a single locus, 16p13.11 (5 case patient deletions: 0 control deletions, p = .08 in the current study, p = .025 compared to published controls). Three 16p13.11 deletions were confirmed de novo, providing further support for the etiological significance of this region. The overall OCD de novo rate was 1.4%, which is intermediate between published rates in controls (0.7%) and in individuals with autism or schizophrenia (2-4%). Several converging lines of evidence implicate 16p13.11 deletions in OCD, with weaker evidence for a role in TS. The trend toward increased overall neurodevelopmental CNV burden in TS and OCD suggests that deletions previously associated with other neurodevelopmental disorders may also contribute to these phenotypes.


[Referral and delivery outcomes in Kivu, Democratic Republic of Congo].
The process of referral between the first and the second level of the health system in the Democratic Republic of Congo is poorly understood. This report intends to study the association between the referral and the hospital perinatal outcomes. Delivery outcomes in a retrospective cohort of 1162 women admitted between June 95 and May 96, in two referral hospitals in Kivu were analyzed according to the referral status and the women's characteristics. Forty-three percent (n=492)of women admitted, corresponding to 2.3% of expected pregnant women, were referred. Referred women had higher risks of obstetrical complications (OR=2.0; CI95%: 1.3-3.1) and intervention (OR=1.5; CI95%: 1.0-2.3) and similar risks of low birth weight and perinatal mortality. Women with complications during the antenatal period had a double risk of intervention and perinatal mortality. The risk of obstetrical intervention was lower when women had attended 2 visits (OR=0.5; CI95%: 0.3-0.8); the risk of low birth weight was lowest only for mothers who had attended one visit (OR=0.5; CI95%: 0.3-0.9). Distance > or =90 minutes walking from home to hospital raised the risk of obstetrical complication (OR=1.7; CI95%: 1.1-2.5), the risk of obstetrical intervention (OR=1.5; CI95%: 1.0-2.1), and the risk of perinatal mortality (OR=1.6; CI95%: 1.0-2.7). Late admission raised the risk of perinatal mortality (OR=1.8; CI95%: 1.2-2.9) and lowered the risk of obstetrical complication (OR=0.7; CI95%: 0.5-1.0). Part payment of care was associated with higher risks of low birth weight (OR=1.9; CI95%: 1.3-2.9), perinatal mortality (OR=2.2; CI95%: 1.4-3.5) and obstetrical intervention (OR=2.4; CI95%: 1.7-3.4). These results suggest a deficit of referred cases considering that 15% of pregnant women in the area covered by the referral hospitals should have been referred. They confirm the negative influence of economic and geographic constraints on the delivery outcomes. They point out the relevance of making reorganization of the referral system a priority.


Supramolecular influence on cis-trans isomerization probed by ion mobility spectrometry.
We used tandem ion mobility spectrometry measurements to investigate how the photo-isomerization of a chromophore (a methylpyridinium derivative) is affected by the complexation with a crown ether. A dramatic blue-shift of the photo-isomerization spectrum was observed upon complexation, which could be well reproduced by ab initio calculations. Our results support that the observed changes in the photo-physical properties of the chromophore originate from the charge-solvation of its pyridinium moiety by the host cage.


A RecQ Protein-like 5 Haplotype is Associated With Colon Cancer.
Emerging murine model data suggests RecQ protein-like 5 (RECQL5) is a tumor suppressor gene. The goal of our study was to test whether RECQL5 gene variants are associated with colon cancer susceptibility. We examined the association of two haplotype-tagging SNPs in RECQL5 and colon cancer in a population-based study of 390 colon cancer cases and 464 population controls. While both crude and covariate-adjusted single SNP analyses were only suggestive for an association with borderline significance (p = 0.07), haplotype analysis shows that individuals carrying the T-G haplotype (rs820196 common allele and rs4789223 minor allele) were at significantly increased risk for colon cancer (OR = 1.34, 95% CI = 1.02-1.76, p = 0.05). Adjustment for age, sex, body mass index, non-steroidal anti-inflammatory use and family history of colon cancer did not alter the results. These results suggest that a haplotype harboring the minor allele of rs4789223 is associated with colon cancer risk. Further study of RECQL5 as a colon cancer susceptibility gene is warranted, particularly with respect to variants in linkage disequilibrium with rs4789223.


Is it mere distraction? Peri-traumatic verbal tasks can increase analogue flashbacks but reduce voluntary memory performance.
Several experiments have shown that we can reduce the frequency of analogue flashbacks with competing tasks presented during a trauma film (i.e. peri-traumatically). A "distraction" hypothesis suggests that any competing task may reduce flashbacks due to distraction and/or a load on executive control. Alternatively, a "modality" hypothesis based on clinical models of PTSD suggests that certain tasks will not protect against intrusions (Experiment 1) and could actually increase them (Experiment 2). Experiment 1 contrasted two concurrent tasks, Verbal Interference (counting backwards in threes) and Visuospatial tapping, against a no-task Control condition during trauma film viewing. The Visuospatial group had significantly fewer intrusions of the film over 1-week than the Control group. Contrary to a distraction account, the Verbal Interference group did not show this effect. Using a larger sample, Experiment 2 showed that the Verbal Interference group (counting backwards in sevens) had more intrusions (and inferior voluntary memory) than no-task Controls. We propose that this is in line with a modality hypothesis concerning trauma flashbacks. Disrupting verbal/conceptual processing during trauma could be harmful for later flashbacks.


Evaluating a systematic voiding programme for patients with urinary incontinence after stroke in secondary care using soft systems analysis and Normalisation Process Theory: findings from the ICONS case study phase.
Urinary incontinence (UI) affects between 40 and 60% of people in hospital after stroke, but is often poorly managed in stroke units. To inform an exploratory trial by three methods: identifying the organisational context for embedding the SVP; exploring health professionals' views around embedding the SVP and measuring presence/absence of UI and frequency of UI episodes at baseline and six weeks post-stroke. A mixed methods single case study included analysis of organisational context using interviews with clinical leaders analysed with soft systems methodology, a process evaluation using interviews with staff delivering the intervention and analysed with Normalisation Process Theory, and outcome evaluation using data from patients receiving the SVP and analysed using descriptive statistics. An 18 bed acute stroke unit in a large Foundation Trust (a 'not for profit' privately controlled entity not accountable to the UK Department of Health) serving a population of 370,000. Health professionals and clinical leaders with a role in either delivering the SVP or linking with it in any capacity were recruited following informed consent. Patients were recruited meeting the following inclusion criteria: aged 18 or over with a diagnosis of stroke; urinary incontinence (UI) as defined by the International Continence Society; conscious; medically stable as judged by the clinical team and with incontinence classified as stress, urge, mixed or 'functional'. All patients admitted to the unit during the intervention period were screened for eligibility; informed consent to collect baseline and outcome data was sought from all eligible patients. Organisational context: 18 health professionals took part in four group interviews. Findings suggest an environment not conducive to therapeutic continence management and a focus on containment of UI. Embedding the SVP into practice: 21 nursing staff took part in six group interviews. Initial confusion gave way to embedding of processes facilitated by new routines and procedures. Patient outcome: 43 patients were recruited; 28 of these commenced the SVP. Of these, 6/28 (21%) were continent at six weeks post-stroke or discharge. It was possible to embed the SVP into practice despite an organisational context not conducive to therapeutic continence care. Recommendations are made for introducing the SVP in a trial context.


Transoral robotic thyroid surgery.
There is currently significant demand for minimally invasive thyroid surgery; however the majority of proposed surgical approaches necessitate a compromise between minimal tissue dissection with a visible cervical scar or extensive tissue dissection with a remote, hidden scar. The development of transoral endoscopic thyroid surgery however provides an approach which is truly minimally invasive, as it conceals the incision within the oral cavity without significantly increasing the amount of required dissection. The transoral endoscopic approach however presents multiple technical challenges, which could be overcome with the incorporation of a robotic operating system. This manuscript summarizes the literature on the feasibility and current clinical experience with transoral robotic thyroid surgery.


Enhanced recovery from ischemia-reperfusion injury in PI3Kα dominant negative hearts: investigating the role of alternate PI3K isoforms, increased glucose oxidation and MAPK signaling.
Classical ischemia-reperfusion (IR) preconditioning relies on phosphatidylinositol 3-kinase (PI3K) for protective signaling. Surprisingly, inhibition of PI3Kα activity using a dominant negative (DN) strategy protected the murine heart from IR injury. It has been proposed that increased signaling through PI3Kγ may contribute to the improved recovery of PI3KαDN hearts following IR. To investigate the mechanism by which PI3KαDN hearts are protected from IR injury, we created a double mutant (PI3KDM) model by crossing p110γ(-/-) (PI3KγKO) with cardiac-specific PI3KαDN mice. The PI3KDM model has morphological and hemodynamic features that are characteristic of both PI3Kγ(-/-) and PI3KαDN mice. Interestingly, when subjected to IR using ex vivo Langendorff perfusion, PI3KDM hearts showed significantly enhanced functional recovery when compared to wildtype (WT) hearts. However, signaling downstream of PI3K through Akt and GSK3β, which has been associated with IR protection, was reduced in PI3KDM hearts. Using ex vivo working heart perfusion, we found no difference in functional recovery after IR between PI3KDM and PI3KαDN; also, glucose oxidation rates were significantly increased in PI3KαDN hearts when compared to WT, and this metabolic shift has been associated with enhanced IR recovery. However, we found that PI3KαDN hearts still had enhanced recovery when perfused exclusively with fatty acids (FA). We then investigated parallel signaling pathways, and found that mitogen-activated protein kinase signaling was increased in PI3KαDN hearts, possibly through the inhibition of negative feedback loops downstream of PI3Kα.


Endocrine abnormalities in HIV-infected women are associated with peak viral load - the Children and Women: AntiRetrovirals and Markers of Aging (CARMA) Cohort.
To investigate the prevalence of endocrine disturbances in a group of HIV-positive (HIV+) women and to identify factors affecting presence of these disorders. To examine specifically whether cellular ageing, as measured by leukocyte telomere length (LTL), is correlated with the presence of endocrine disturbance. A cross-sectional retrospective substudy of an ongoing prospective cohort study. Adult HIV+ (≥19 years) women enrolled in the CARMA (Children and Women: AntiRetrovirals and Markers of Aging) cohort study (N = 192). Prevalences of T2DM, glucose intolerance, dyslipidaemia, thyroid disorders, adrenal insufficiency, hypogonadism, primary ovarian insufficiency (POI), demographics, HIV and hepatitis C virus (HCV) infection status, baseline LTL, combined antiRetroviral therapy (cART) and substance exposures were collected. Statistical analysis included univariable followed by multivariable Poisson regression and step-wise reduction to refine the multivariable model. Prevalence of any endocrine abnormality was 58% (dyslipidaemia 43%, glucose intolerance/T2DM 13%, thyroid disorders 15%). In multivariable analysis, age was associated with number and type (any, glucose, lipid) of abnormality, while increasing body mass index (BMI) was associated with number of diagnoses and with glucose metabolism disorders. Interestingly, peak HIV pVL ≥100 000 copies/ml was associated with any abnormality, total number of disorders and presence of a thyroid disorder, while any disorder, glucose abnormalities and dyslipidaemia were negatively associated with alcohol use. LTL was not associated with number or type of endocrine abnormalities in this study. Further studies examining the relationship between duration and extent of exposure to HIV viraemia in relation to developing abnormal endocrine function are warranted.


Hypothalamic-Pituitary-Adrenal Axis Activity in Childhood Predicts Emotional Memory Effects and Related Neural Circuitry in Adolescent Girls.
Negative emotional experiences can be more difficult to forget than neutral ones, a phenomenon termed the "emotional memory effect." Individual differences in the strength of the emotional memory effect are associated with emotional health. Thus, understanding the neurobiological underpinnings of the emotional memory effect has important implications, especially for individuals at risk for emotional health problems. Although the neural basis of emotional memory effects has been relatively well defined, less is known about how hormonal factors that can modulate emotional memory, such as glucocorticoids, relate to that neural basis. Importantly, probing the role of glucocorticoids in the stress- and emotion-sensitive period of late childhood to adolescence could provide actionable points of intervention. We addressed this gap by testing whether hypothalamic-pituitary-adrenal (HPA) axis activity during a parent-child conflict task at 11 years of age predicted emotional memory and its primary neural circuitry (i.e., amygdala-hippocampus functional connectivity) at 16 years of age in a longitudinal study of 147 girls (104 with complete data). Results showed that lower HPA axis activity predicted stronger emotional memory effects, r(124) = -.236, p < .01, and higher emotional memory-related functional connectivity between the right hippocampus and the right amygdala, β = -.385, p < .001. These findings suggest that late childhood HPA axis activity may modulate the neural circuitry of emotional memory effects in adolescence, which may confer a potential risk trajectory for emotional health among girls.


Impairment of endothelial function in patients with multiple sclerosis.
Large epidemiological studies suggest higher risk of vascular events in patients with multiple sclerosis (MS). Chronic inflammatory response and oxidative stress, key-players in a process of atherogenesis, are also suspected to play a role in pathophysiology of MS. Prospective studies elucidating risk of atherosclerosis in MS patients are currently missing. The aim of the study was to assess endothelial function in patients with MS and in healthy controls. We enrolled 46 patients with diagnosis of relapsing-remitting MS and age-matched population of 31 healthy subjects. Endothelial function was assessed using peripheral arterial tonometry and expressed as reperfusion hyperemia index (RHI). RHI in MS population was significantly lower than in controls (1.77 vs 2.30; p=0.001), even though control population seemed to have higher burden of known vascular risk factors (significantly higher portion of male sex and significantly higher body mass index; p ≤ 0.001 for both parameters). The presence of MS was the only significant independent variable associated with the RHI (beta=0.396, p<0.001) in multiple linear regression model. Results of our study suggest significant impairment of endothelial function in MS population compared to age matched control population with low burden of vascular risk factors.


Mapping open knowledge institutions: an exploratory analysis of Australian universities.
While the movement for open research has gained momentum in recent years, there remain concerns about the broader commitment to openness in knowledge production and dissemination. Increasingly, universities are under pressure to transform themselves to engage with the wider community and to be more inclusive. Open knowledge institutions (OKIs) provide a framework that encourages universities to act with the principles of openness at their centre; not only should universities embrace digital open access (OA), but also lead actions in cultivating diversity, equity, transparency and positive changes in society. This leads to questions of whether we can evaluate the progress of OKIs and what are potential indicators for OKIs. As an exploratory study, this article reports on the collection and analysis of a list of potential OKI indicators. Data for these indicators are gathered for 43 Australian universities. The indicators provide high-dimensional and complex signals about university performances. They show evidence of large disparities in characteristics such as Indigenous employment and gender equity, and a preference for repository-mediated OA across Australian universities. We demonstrate use of the OKI evaluation framework to categorise these indicators into three platforms of diversity, communication and coordination. The analysis provides new insights into the Australian open knowledge landscape and ways of mapping different paths of OKIs.


Wireless Synthesis and Activation of Electrochemiluminescent Thermoresponsive Janus Objects Using Bipolar Electrochemistry.
In this work, bipolar electrochemistry (BPE) is used as a dual wireless tool to generate and to activate a thermoresponsive electrochemiluminescent (ECL) Janus object. For the first time, BPE allows regioselective growth of a poly(N-isopropylacrylamide) (pNIPAM) hydrogel film on one side of a carbon fiber. It is achieved thanks to the local reduction of persulfate ions, which initiate radical polymerization of NIPAM. By controlling the electric field and the time of the bipolar electrochemical reactions, we are able to control the length and the thickness of the deposit. The resulting pNIPAM film is found to be swollen in water at room temperature and collapsed when heated above 32 °C. We further incorporated a covalently attached ruthenium complex luminophore, Ru(bpy)32+, in the hydrogel film. In the second time, BPE is used to activate remotely the electrogenerated chemiluminescence (ECL) of the Ru(bpy)32+ moieties in the film. We take advantage of the film responsiveness to amplify the ECL signal. Upon collapse of the film, the ECL signal, which is sensitive to the distance between adjacent Ru(bpy)32+ centers, is strongly amplified. It is therefore shown that BPE is a versatile tool to generate highly sophisticated materials based on responsive polymers, which could lead to sensitive sensors.


Effects of T antigen and replication protein A on the initiation of DNA synthesis by DNA polymerase alpha-primase.
Studies of simian virus 40 (SV40) DNA replication in a reconstituted cell-free system have established that T antigen and two cellular replication proteins, replication protein A (RP-A) and DNA polymerase alpha-primase complex, are necessary and sufficient for initiation of DNA synthesis on duplex templates containing the SV40 origin of DNA replication. To better understand the mechanism of initiation of DNA synthesis, we analyzed the functional interactions of T antigen, RP-A, and DNA polymerase alpha-primase on model single-stranded DNA templates. Purified DNA polymerase alpha-primase was capable of initiating DNA synthesis de novo on unprimed single-stranded DNA templates. This reaction involved the synthesis of a short oligoribonucleotide primer which was then extended into a DNA chain. We observed that the synthesis of ribonucleotide primers by DNA polymerase alpha-primase is dramatically stimulated by SV40 T antigen. The presence of T antigen also increased the average length of the DNA product synthesized on primed and unprimed single-stranded DNA templates. These stimulatory effects of T antigen required direct contact with DNA polymerase alpha-primase complex and were most marked at low template and polymerase concentrations. We also observed that the single-stranded DNA binding protein, RP-A, strongly inhibits the primase activity of DNA polymerase alpha-primase, probably by blocking access of the enzyme to the template. T antigen partially reversed the inhibition caused by RP-A. Our data support a model in which DNA priming is mediated by a complex between T antigen and DNA polymerase alpha-primase with the template, while RP-A acts to suppress nonspecific priming events.


Higher Postinduction Infliximab Serum Trough Levels Are Associated With Healing of Fistulizing Perianal Crohn's Disease in Children.
There is some evidence in adults that higher serum infliximab (IFX) levels are needed to adequately treat fistulizing perianal Crohn's disease (CD). However, data in children are lacking. We aimed to determine postinduction serum trough IFX levels that are associated with healing of fistulizing perianal CD (PCD) at week 24. In a multicenter inception cohort study, consecutive children younger than age 17 years with fistulizing perianal CD treated with IFX between April 2014 and June 2017 who had serum trough IFX titers measured before the fourth infusion were included. Area under the receiver operating characteristic curve (AUROC) was calculated to determine the best cutoff to predict fistula healing. A total of 667 children with Crohn's disease were recruited, with 85 (12.7%) patients diagnosed with fistulizing PCD. There were 27 of 52 (52%) children in whom pre-fourth infusion IFX levels were measured (mean age, 12.57 ± 5.12 years). At week 24, 14 of 27 (52%) patients responded with healing/healed PCD, whereas the rest had ongoing active fistulizing disease. The median IFX pre-fourth dose level in the responders was 12.7 ug/mL, compared with 5.4 ug/mL in the active disease group (P = 0.02). There was a strong correlation between IFX levels and healing of fistulizing PCD at week 24 (r = 0.65; P < 0.001). The AUROC was 0.80 (95% confidence interval, 0.64-0.97; P = 0.007) for pre-fourth IFX level to predict response of fistulizing PCD at week 24, and a level of 12.7 ug/mL best predicted fistula healing. Higher trough IFX levels are associated with healing of fistulizing perianal CD.


Resisting global AIDS knowledges: born-again Christian narratives of the epidemic from Papua New Guinea.
The recognition that HIV prevention materials need to be adapted to local cultures is not often sufficiently understood and applied. Counter discourses and determined disputation about the best means of HIV prevention show that success is not simply a matter of mindfully translating globally sanctioned knowledge and presenting it to receptive audiences. Beliefs contrary to global AIDS knowledges will not be displaced inevitably by scientific facts. As this study of born-again Christians in Papua New Guinea shows, there is incommensurability between the globalized approach preferred by the government and the approach of these Christians. The answer may lie in two words: respect and dialogue.


[Bacterial translocation and its surgical implications].
The passage of viable bacteria through epithelial mucosa into lamina propria and then to mesentheric lymphnodes, and possibly other tissues was defined bacterial translocation (BT) by Berg and Garlington. The transepithelial passage of bacteria out of the intestinal lumen seems to occur in a variety of surgical conditions, and it is suspected to be involved in systemic inflammatory response syndrome, sepsis and multiorgan failure. There is increasing evidence that many nosocomial infections are caused by enteric translocating microorganisms, although the exact incidence of BT in humans is difficult to establish. Consensus is emerging that the barrier function of the gut is relevant in established critical illness and in patients at risk of developing sepsis and clinical studies have provided strong evidence that systemic infections often originate from intestinal flora in high risk patients. So the suspect of BT is made when there is infection after trauma, burns, major surgery, chemotherapy and immuno suppression. Bacterial translocation is also associated with organ transplantation, especially with small bowel transplantation. The Authors have summarized published experimental and clinical studies that have tried to understand the occurrence, mechanisms and effects of this complex process. At the present time there is a near full understanding of the relevance of BT like an interplay of diverse factors in a physiologically, immunologically and microbiologically complex intestinal tract. However additional experimental and clinical studies are needed to clarify the relationship between these phenomena and the development of sepsis or multiple organ dysfunction syndrome.


Cranberry, D-mannose and anti-inflammatory agents prevent lower urinary tract symptoms in women undergoing prolapse surgery.
Objective: We assessed the effect on lower urinary tract symptoms (LUTS) of a supplement containing cranberry, D-mannose and anti-inflammatory molecules in postmenopausal women undergoing surgery for cystocele.Study design: Forty postmenopausal women were randomized 1:1 to an active group receiving the nutritional supplement twice a day for 2 weeks starting from surgery, or to a control group receiving surgery only. Primary outcomes were the effectiveness in the postoperative LUTS and urinary tract infections (UTI). LUTS were investigated by a validated questionnaire (ICIQ-FLUTS) at baseline and at week 4. Secondary outcomes were the safety and tolerability of the supplement and other perioperative outcomes.Results: No significant differences were found in perioperative outcomes and in incidence of UTI. After surgery, women treated with the supplement experienced significantly better scores on the filling domain of the questionnaire. A non-significant decrease in voiding scores was also found. No adverse events were detected.Conclusion: The use of an oral supplement containing cranberry, D-mannose and anti-inflammatory molecules decreases the perception of LUTS in postmenopausal women after anterior colporraphy. Our data suggest that perioperative use of nutritional supplements may be useful in the management of postoperative LUTS.


Remarkable differences in the biochemical fate of Cd2+, Hg2+, CH3Hg+ and thimerosal in red blood cell lysate.
Humans are environmentally exposed to potentially toxic Cd and Hg species and to the Hg compound thimerosal (THI), an antibactericidal vaccine additive. Previous studies have revealed that Cd2+, Hg2+ and CH3Hg+ are taken up by red blood cells (RBCs) and bind to cytosolic glutathione (GSH) and/or hemoglobin (Hb). Since interactions in the cytosol of RBCs may be linked to their hemolysis, a more comprehensive characterization of these interactions was sought. After the addition of each Cd and Hg species to RBC lysate, the mixtures were analyzed after 5 min, 2 h and 6 h by size-exclusion chromatography (SEC) coupled on-line to an inductively coupled plasma atomic emission spectrometer (ICP-AES). In contrast to previous studies, however, reducing conditions were maintained by employing a 100 mM Tris buffer mobile phase (pH 7.4), which contained ∼2.5 mM of glutathione (GSH). At ≥2 h, ∼85% of Cd2+ weakly interacted with hemoglobin (Hb), while ∼13% eluted as (GS)xCd and ∼2% bound to a ≥70 kDa Cd-binding protein. In contrast, ∼6% of Hg2+ co-eluted with Hb at all time points, while ∼94% eluted as (GS)xHg. The results for CH3Hg+ showed that ∼5% of Hg co-eluted with Hb, while for THI this percentage gradually increased to 12% (6 h). The remaining Hg eluted as GS-HgCH3 and GS-HgCH2CH3. Our results revealed remarkable differences in the interaction of the investigated Cd and Hg species with cytosolic RBC constituents. The formation of (Hb)xHg species, regardless of which Hg compound was added, suggests their mammalian toxicology to be intertwined with the metabolism of Fe.


Coordination of locomotor and cardiorespiratory networks of Lymnaea stagnalis by a pair of identified interneurones.
1. The morphology and electrophysiology of a newly identified bilateral pair of interneurones in the central nervous system of the pulmonate pond snail Lymnaea stagnalis is described. 2. These interneurones, identified as left and right pedal dorsal 11 (L/RPeD11), are electrically coupled to each other as well as to a large number of foot and body wall motoneurones, forming a fast-acting neural network which coordinates the activities of foot and body wall muscles. 3. The left and right sides of the body wall of Lymnaea are innervated by left and right cerebral A cluster neurones. Although these motoneurones have only ipsilateral projections, they are indirectly electrically coupled to their contralateral homologues via their connections with L/RPeD11. Similarly, the activities of left and right pedal G cluster neurones, which are known to be involved in locomotion, are also coordinated by L/RPeD11. 4. Selective ablation of both neurones PeD11 results in the loss of coordination between the bilateral cerebral A clusters. 5. Interneurones L/RPeD11 are multifunctional. In addition to coordinating motoneuronal activity, they make chemical excitatory connections with heart motoneurones. They also synapse upon respiratory motoneurones, hyperpolarizing those involved in pneumostome opening (expiration) and depolarizing those involved in pneumostome closure (inspiration). 6. An identified respiratory interneurone involved in pneumostome closure (visceral dorsal 4) inhibits L/RPeD11 together with all their electrically coupled follower cells. 7. Both L/RPeD11 have strong excitatory effects on another pair of electrically coupled neurones, visceral dorsal 1 and right parietal dorsal 2, which have previously been shown to be sensitive to changes in the partial pressure of environmental oxygen (PO2). 8. Although L/RPeD11 participate in whole-body withdrawal responses, electrical stimulation applied directly to these neurones was not sufficient to induce this behaviour.


Simultaneous Spray Drying for Combination Dry Powder Inhaler Formulations.
Spray drying is a particle engineering technique used to manufacture respirable pharmaceutical powders that are suitable for delivery to the deep lung. It is amenable to processing both small molecules and biologic actives, including proteins. In this work, a simultaneous spray-drying process, termed simul-spray, is described; the process involves two different active pharmaceutical ingredient (API) solutions that are simultaneously atomized through separate nozzles into a single-spray dryer. Collected by a single cyclone, simul-spray produces a uniform mixture of two different active particles in a single-unit operation. While combination therapies for dry powder inhalers containing milled small molecule API are commercially approved, limited options exist for preparing combination treatments that contain both small molecule APIs and biotherapeutic molecules. Simul-spray drying is also ideal for actives which cannot withstand a milling-based particle engineering process, or which require a high dose that is incompatible with a carrier-based formulation. Three combination case studies are demonstrated here, in which bevacizumab is paired with erlotinib, cisplatin, or paclitaxel in a dry powder inhaler formulation. These model systems were chosen for their potential relevance to the local treatment of lung cancer. The resulting formulations preserved the biologic activity of the antibody, achieved target drug concentration, and had aerosol properties suitable for pulmonary delivery.


Prevalence of unintended pregnancy and its associated factors: Evidence from six south Asian countries.
Unintended pregnancy is a significant public health concern in South Asian countries because of its negative association with the socioeconomic and health outcomes for both children and mothers. The present study aimed to explore the prevalence of unintended pregnancy and explore its determinants among women of reproductive age in six South Asian countries. Nationwide latest demography and health survey data from six South Asian countries, including Bangladesh (2014), Pakistan (2017-2018), Nepal (2016), Afghanistan (2015), Maldives (2016-2017) and India (2015-2016) were pooled for the present study. Multivariate analysis was performed to explore the association between unintended pregnancy and its associated factors. Amongst the total women (n = 41,689), overall, 19.1% pregnancies were reported as unintended (ranging from 11.9% in India to 28.4% in Bangladesh). The logistic regression model showed that younger women (15-19 years) had 1.42 times higher chance of unintended pregnancies. The odds of unintended pregnancies was 1.24 times higher for poorest women and 1.19 times higher for poorer women. Further, urban women (aOR = 0.70, 95% CI = 0.50-0.80), women having no children (aOR = 0.10, 95% CI = 0.09-0.12), smaller (≤4) family (aOR = 0.72, 95% CI = 0.67-0.78), those who intent to use contraceptive (aOR = 0.72, 95% CI = 0.60-0.86), currently living with partner (aOR = 0.90, 95% CI = 0.81-0.99), first cohabitation in teenage (≤19 years) (aOR = 0.85, 95% CI = 0.78-0.92) were less likely to report unintended pregnancies. This study has showed that women's age, wealth index, place of residence, number of children, family size, the intention of contraceptive use, living with a partner, and first cohabitation age are essential determinants of unintended pregnancy. These factors should be considered when trying to reduce unintended pregnancy in six South Asian countries. However, there is a need to improve health education, counselling, skills-building, sex education, modern contraceptive use and its access in this region. Intervention programs regarding reproductive health and policies are warranted to reduce rates of unintended pregnancy in South Asian countries.


Duration of shedding of respiratory syncytial virus in a community study of Kenyan children.
Our understanding of the transmission dynamics of respiratory syncytial virus (RSV) infection will be better informed with improved data on the patterns of shedding in cases not limited only to hospital admissions. In a household study, children testing RSV positive by direct immunofluorescent antibody test (DFA) were enrolled. Nasal washings were scheduled right away, then every three days until day 14, every 7 days until day 28 and every 2 weeks until a maximum of 16 weeks, or until the first DFA negative RSV specimen. The relationship between host factors, illness severity and viral shedding was investigated using Cox regression methods. From 151 families a total of 193 children were enrolled with a median age of 21 months (range 1-164 months), 10% infants and 46% male. The rate of recovery from infection was 0.22/person/day (95% CI 0.19-0.25) equivalent to a mean duration of shedding of 4.5 days (95%CI 4.0-5.3), with a median duration of shedding of 4 days (IQR 2-6, range 1-14). Children with a history of RSV infection had a 40% increased rate of recovery i.e. shorter duration of viral shedding (hazard ratio 1.4, 95% CI 1.01-1.86). The rate of cessation of shedding did not differ significantly between males and females, by severity of infection or by age. We provide evidence of a relationship between the duration of shedding and history of infection, which may have a bearing on the relative role of primary versus re-infections in RSV transmission in the community.


The effect of fiscal policy on diet, obesity and chronic disease: a systematic review.
To assess the effect of food taxes and subsidies on diet, body weight and health through a systematic review of the literature. We searched the English-language published and grey literature for empirical and modelling studies on the effects of monetary subsidies or taxes levied on specific food products on consumption habits, body weight and chronic conditions. Empirical studies were dealing with an actual tax, while modelling studies predicted outcomes based on a hypothetical tax or subsidy. Twenty-four studies met the inclusion criteria: 13 were from the peer-reviewed literature and 11 were published on line. There were 8 empirical and 16 modelling studies. Nine studies assessed the impact of taxes on food consumption only, 5 on consumption and body weight, 4 on consumption and disease and 6 on body weight only. In general, taxes and subsidies influenced consumption in the desired direction, with larger taxes being associated with more significant changes in consumption, body weight and disease incidence. However, studies that focused on a single target food or nutrient may have overestimated the impact of taxes by failing to take into account shifts in consumption to other foods. The quality of the evidence was generally low. Almost all studies were conducted in high-income countries. Food taxes and subsidies have the potential to contribute to healthy consumption patterns at the population level. However, current evidence is generally of low quality and the empirical evaluation of existing taxes is a research priority, along with research into the effectiveness and differential impact of food taxes in developing countries.


Open-source, Python-based, hardware and software for controlling behavioural neuroscience experiments.
Laboratory behavioural tasks are an essential research tool. As questions asked of behaviour and brain activity become more sophisticated, the ability to specify and run richly structured tasks becomes more important. An increasing focus on reproducibility also necessitates accurate communication of task logic to other researchers. To these ends, we developed pyControl, a system of open-source hardware and software for controlling behavioural experiments comprising a simple yet flexible Python-based syntax for specifying tasks as extended state machines, hardware modules for building behavioural setups, and a graphical user interface designed for efficiently running high-throughput experiments on many setups in parallel, all with extensive online documentation. These tools make it quicker, easier, and cheaper to implement rich behavioural tasks at scale. As important, pyControl facilitates communication and reproducibility of behavioural experiments through a highly readable task definition syntax and self-documenting features. Here, we outline the system's design and rationale, present validation experiments characterising system performance, and demonstrate example applications in freely moving and head-fixed mouse behaviour.


Intrinsic and synaptic properties of hippocampal CA1 pyramidal neurons of the Wistar Audiogenic Rat (WAR) strain, a genetic model of epilepsy.
Despite the many studies focusing on epilepsy, a lot of the basic mechanisms underlying seizure susceptibility are mainly unclear. Here, we studied cellular electrical excitability, as well as excitatory and inhibitory synaptic neurotransmission of CA1 pyramidal neurons from the dorsal hippocampus of a genetic model of epilepsy, the Wistar Audiogenic Rat (WARs) in which limbic seizures appear after repeated audiogenic stimulation. We examined intrinsic properties of neurons, as well as EPSCs evoked by Schaffer-collateral stimulation in slices from WARs and Wistar parental strain. We also analyzed spontaneous IPSCs and quantal miniature inhibitory events. Our data show that even in the absence of previous seizures, GABAergic neurotransmission is reduced in the dorsal hippocampus of WARs. We observed a decrease in the frequency of IPSCs and mIPSCs. Moreover, mIPSCs of WARs had faster rise times, indicating that they probably arise from more proximal synapses. Finally, intrinsic membrane properties, firing and excitatory neurotransmission mediated by both NMDA and non-NMDA receptors are similar to the parental strain. Since GABAergic inhibition towards CA1 pyramidal neurons is reduced in WARs, the inhibitory network could be ineffective to prevent the seizure-dependent spread of hyperexcitation. These functional changes could make these animals more susceptible to the limbic seizures observed during the audiogenic kindling.


From what might have been to what must have been: counterfactual thinking creates meaning.
Four experiments explored whether 2 uniquely human characteristics-counterfactual thinking (imagining alternatives to the past) and the fundamental drive to create meaning in life-are causally related. Rather than implying a random quality to life, the authors hypothesized and found that counterfactual thinking heightens the meaningfulness of key life experiences. Reflecting on alternative pathways to pivotal turning points even produced greater meaning than directly reflecting on the meaning of the event itself. Fate perceptions ("it was meant to be") and benefit-finding (recognition of positive consequences) were identified as independent causal links between counterfactual thinking and the construction of meaning. Through counterfactual reflection, the upsides to reality are identified, a belief in fate emerges, and ultimately more meaning is derived from important life events.


Controlling firearms use in Australia: has the 1996 gun law reform produced the decrease in rates of suicide with this method?
Observed reductions in firearm suicides in Australia have been linked to the 1997 national firearms agreement (NFA) introduced following the 1996 Port Arthur massacre. The NFA placed strong access restrictions on firearms. To assess the impact of legislative restrictions on the incidence of firearm suicide in Queensland and explore alternative or contributory factors behind observed declines. The Queensland suicide register (QSR) provided detailed information on all male suicides in Queensland (1990-2004), with additional data for Australia (1968-2004) accessed from other official sources. Trends in suicide rates pre/post NFA, and in method selection, were assessed using negative binomial regressions. Changing method selection patterns were examined using a cohort analysis of 5 years of age classes for Australian males. The observed reduction in firearms suicides was initiated prior to the 1997 introduction of the NFA in Queensland and Australia, with a clear decline observed in Australian figures from 1988. No significant difference was found in the rate pre/post the introduction of the NFA in Queensland; however, a significant difference was found for Australian data, the quality of which is noticeably less satisfactory. A marked age-difference in method choice was observed through a cohort analysis demonstrating both time and age influences. Within sequential birth cohorts, rates of firearms suicides decreased in younger males but increased in hanging suicides; this trend was far less marked in older males. The implemented restrictions may not be responsible for the observed reductions in firearms suicide. Data suggest that a change in social and cultural attitudes could have contributed to the shift in method preference.


NGF improves spatial memory in aged rodents as a function of age.
Aged rats were tested for place navigation in a circular water maze for spatial memory ability at 18 and 30 months of age; 45% of the 18-month-old rats displayed impaired place navigation performance relative to young control rats, while essentially all of the 30-month-old rats were impaired. The aged impaired rats were retested twice during NGF or vehicle infusion in the right lateral ventricle. In the 18-month-old group, NGF-infused rats showed improved retention of previously acquired place navigation performance and improved spatial acuity over the former platform site when the invisible platform was removed. NGF infusion also had a significant effect in the much more severely impaired 30-month-old rats: while the vehicle-infused aged rats showed a progressive decline in the performance between the first and second test weeks, the performance of the NGF-infused rats remained stable throughout the infusion period. The interpretation of these effects in the oldest animals, however, was confounded by a progressive decline in swim speed seen in the vehicle-infused animals. The 30-month-old vehicle-infused control rats showed a significant cell loss and cell shrinkage relative to the young control rats in the septal/diagonal band area, the striatum, and the nucleus basalis as assessed by NGF-receptor (NGFr) and ChAT double-label immunocytochemistry. A significant increase in the size but not in the number of cells was observed on the side of the NGF infusion in the 30-month-old NGF-infused rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Disparities in health care affordability among childhood cancer survivors persist following the Affordable Care Act.
The Affordable Care Act (ACA) included many provisions that may have improved access to care for childhood cancer survivors (CCS). In this study, we sought to compare health insurance coverage and the affordability of health care among adult childhood CCS before and after the implementation of the ACA. Using data from the National Health Interview Survey (NHIS), two cohorts of CCS age 21-65 years old and matched (1:3) controls without a history of cancer were identified. A difference-in-differences analysis was used to compare insurance coverage and health care affordability pre- (2011-2013) and post-ACA (2015-2017). There were 309 CCS identified in the pre-ACA cohort and 324 in the post-ACA cohort. The two cohorts were similar in demographic composition. Prior to the ACA, CCS were 39% more likely to be uninsured than their peers (p = .046). Post, there was no difference in the odds of being uninsured between CCS and their peers. Following implementation of the ACA, the proportion of CCS who reported having difficulty with the affordability of health care decreased (p = .013) as did the proportion reporting skipping needed care due to cost (p < .001). However, 13% of CCS still reported being uninsured, 36% reported difficulty paying for health care, and 13% reported foregoing needed care due to cost. Relative to their peers, CCS saw improvement in foregoing needed care due to cost, but disparities still remain. Although improvements were observed, health care affordability, and medical nonadherence remains a problem for CCS. Additional efforts are needed to improve health care affordability among CCS.


Supporting new graduate nurses making the transition to rural nursing practice: views from experienced rural nurses.
To present the findings from the experienced rural nurse participants of a larger study that explored the transitional experiences of newly graduated nurses making the role transition in rural health care facilities in Australia. There are specific and unique aspects of rural nursing practice that influence the nature and timing of support for new graduate nurses that have not been explored or acknowledged as influencing the new graduate nurses' experience of transition. Specifically, the difficulties and challenges that experienced rural nurses face in providing effective and timely support for new graduate nurses who are making the transition to rural nursing practice is yet to be explored. Using a qualitative case study framework, this study specifically aimed to investigate and describe the nature and timing of support required during the transition to nursing practice that is specific for the rural context and capacity. Individual in-depth interviews were conducted with 16 experienced rural nurses who, at the time of the study, worked with new graduate nurses in the rural practice environment. The findings from this study showed that the provision of timely on-ward support for new graduates making the transition to rural nursing practice is affected and influenced by the skill mix and staffing allocation within the rural environment. As well, there is a lack of awareness by rural nurses of how to meet the on-ward support needs of new graduate nurses. This study has identified the specific and unique aspects of the rural nurse's role and responsibilities for which the new graduate nurse requires incremental learning and intensive clinical support. The findings can be used by rural health services and experienced rural registered nurses to assist in implementing adequate and timely support for new graduate nurses.


Direct observation of ice nucleation events on individual atmospheric particles.
Heterogeneous ice nucleation is a physical chemistry process of critical relevance to a range of topics in the fundamental and applied sciences and technologies. Heterogeneous ice nucleation remains insufficiently understood, partially due to the lack of experimental methods capable of obtaining in situ microscopic details of ice formation over nucleating substrates or particles. We present microscopic observations of ice nucleation events on kaolinite particles at the nanoscale and demonstrate the capability of direct tracking and micro-spectroscopic characterization of individual ice nucleating particles (INPs) in an authentic atmospheric sample. This approach utilizes a custom-built ice nucleation cell, interfaced with an Environmental Scanning Electron Microscope (IN-ESEM platform) operated at temperatures and relative humidities relevant for heterogeneous ice nucleation. The IN-ESEM platform allows dynamic observations of individual ice formation events over particles in isobaric and isothermal experiments. Isothermal experiments on individual kaolinite particles demonstrate that ice crystals preferably nucleate at the edges of the stacked kaolinite platelets, rather than on their basal planes. These experimental observations of the location of ice nucleation provide direct information for further theoretical chemistry predictions of ice formation on kaolinite.


Assessing the value of DNA barcodes and other priority gene regions for molecular phylogenetics of Lepidoptera.
Despite apparently abundant amounts of observable variation and species diversity, the order Lepidoptera exhibits a morphological homogeneity that has provided only a limited number of taxonomic characters and led to widespread use of nucleotides for inferring relationships. This study aims to characterize and develop methods to quantify the value of priority gene regions designated for Lepidoptera molecular systematics. In particular, I assess how the DNA barcode segment of the mitochondrial COI gene performs across a broad temporal range given its number one position of priority, most sequenced status, and the conflicting opinions on its phylogenetic performance. Gene regions commonly sequenced for lepidoptera phylogenetics were scored using multiple measures across three categories: practicality, which includes universality of primers and sequence quality; phylogenetic utility; and phylogenetic signal. I found that alternative measures within a category often appeared correlated, but high scores in one category did not necessarily translate into high scores in another. The DNA barcode was easier to sequence than other genes, and had high scores for utility but low signal above the genus level. Given limited financial resources and time constraints, careful selection of gene regions for molecular phylogenetics is crucial to avoid wasted effort producing partially informative data. This study introduces an approach to assessing the value of gene regions prior to the initiation of new studies and presents empirical results to help guide future selections.


Risk factors for unstructured treatment interruptions and association with survival in low to middle income countries.
Antiretroviral therapy (ART) treatment interruptions lead to poor clinical outcomes with unplanned or unstructured TIs (uTIs) likely to be underreported. This study describes; uTIs, their risk factors and association with survival. Analysis of ART programmatic data from 11 countries across Asia and Africa between 2003 and 2013 where an uTI was defined as a ≥90-day patient initiated break from ART calculated from the last day the previous ART prescription would have run out until the date of the next ART prescription. Factors predicting uTI were assessed with a conditional risk-set multiple failure time-to-event model to account for repeated events per subject. Association between uTI and mortality was assessed using Cox proportional hazards, with a competing risks extension to test for the influence of lost to follow-up (LTFU). 40,632 patients were included from 11 countries across 33 sites (17 Africa, 16 Asia). Median duration of follow-up was 1.61 years (IQR 0.54-3.31 years), 3386 (8.3 %) patients died, and 3453 (8.5 %) were LTFU. There were 14,817 uTIs, with 10,162 (25 %) patients having more than one uTI. In the adjusted model males were at lower risk of uTI (aHR 0.94, p < 0.01, and age 20-59 was protective compared to <20 years (20-39 years aHR 0.87, p < 0.01; 40-59 years aHR 0.86, p < 0.01). Preserved immune function, as measured by higher CD4 cell count, was associated with a reduced rate of uTI compared to CD4 <200 cells/μL (CD4 200-350 cells/μL aHR 0.89, p < 0.01; CD4 >350 cells/μL aHR 0.87, p < 0.01), whereas advanced clinical disease was associated with increased uTI rate (WHO stage 3 aHR 1.10, p < 0.01; WHO stage 4 aHR 1.21, p < 0.01). There was no relationship between uTI and mortality after adjusting for disease status and considering LTFU as a competing risk. uTIs were frequent in people in ART programs in low-middle income countries and associated with younger age, female gender and advanced HIV. uTI did not predict survival when loss to follow-up was considered a competing risk. Further evaluation of uTI predictors and interventions to reduce their occurrence is warranted.


Association analysis of loci implied in "buffering" epistasis.
The existence of buffering mechanisms is an emerging property of biological networks, and this results in the buildup of robustness through evolution. So far, there are no explicit methods to find loci implied in buffering mechanisms. However, buffering can be seen as interaction with genetic background. Here we develop this idea into a tractable model for quantitative genetics, in which the buffering effect of one locus with many other loci is condensed into a single statistical effect, multiplicative on the total additive genetic effect. This allows easier interpretation of the results and simplifies the problem of detecting epistasis from quadratic to linear in the number of loci. Using this formulation, we construct a linear model for genome-wide association studies that estimates and declares the significance of multiplicative epistatic effects at single loci. The model has the form of a variance components, norm reaction model and likelihood ratio tests are used for significance. This model is a generalization and explanation of previous ones. We test our model using bovine data: Brahman and Tropical Composite animals, phenotyped for body weight at yearling and genotyped at high density. After association analysis, we find a number of loci with buffering action in one, the other, or both breeds; these loci do not have a significant statistical additive effect. Most of these loci have been reported in previous studies, either with an additive effect or as footprints of selection. We identify buffering epistatic SNPs present in or near genes reported in the context of signatures of selection in multi-breed cattle population studies. Prominent among these genes are those associated with fertility (INHBA, TSHR, ESRRG, PRLR, and PPARG), growth (MSTN, GHR), coat characteristics (KIT, MITF, PRLR), and heat resistance (HSPA6 and HSPA1A). In these populations, we found loci that have a nonsignificant statistical additive effect but a significant epistatic effect. We argue that the discovery and study of loci associated with buffering effects allow attacking the difficult problems, among others, of the release of maintenance variance in artificial and natural selection, of quick adaptation to the environment, and of opposite signs of marker effects in different backgrounds. We conclude that our method and our results generate promising new perspectives for research in evolutionary and quantitative genetics based on the study of loci that buffer effect of other loci.


Implementing new health interventions in developing countries: why do we lose a decade or more?
It is unclear how long it takes for health interventions to transition from research and development (R&D) to being used against diseases prevalent in resource-poor countries. We undertook an analysis of the time required to begin implementation of four vaccines and three malaria interventions. We evaluated five milestones for each intervention, and assessed if the milestones were associated with beginning implementation. The authors screened World Health Organization (WHO) databases to determine the number of years between first regulatory approval of interventions, and countries beginning implementation. Descriptive analyses of temporal patterns and statistical analyses using logistic regression and Cox proportional hazard models were used to evaluate associations between five milestones and the beginning of implementation for each intervention. The milestones were: (A) presence of a coordinating group focused on the intervention; (B) availability of an intervention tailored to developing country health systems; (C) international financing commitment, and; (D) initial and (E) comprehensive WHO recommendations. Countries were categorized by World Bank income criteria. Five years after regulatory approval, no low-income countries (LICs) had begun implementing any of the vaccines, increasing to an average of only 4% of LICs after 10 years. Each malaria intervention was used by an average of 7% of LICs after five years and 37% after 10 years. Four of the interventions had similar implementation rates to hepatitis B vaccine (HepB), while one was slower and one was faster than HepB. A financing commitment and initial WHO recommendation appeared to be temporally associated with the beginning of implementation. The initial recommendation from WHO was the only milestone associated in all statistical analyses with countries beginning implementation (relative rate = 1.97, P < 0.001). Although possible that four milestones were not associated with countries beginning implementation, we propose an alternative interpretation; that the milestones were not realized early enough in each intervention's development to shorten the time to beginning implementation. We discuss a framework built upon existing literature for consideration during the development of future interventions. Identifying critical milestones and their timing relative to R&D, promises to help new interventions realize their intended public health impact more rapidly.


Iterative Supervised Principal Component Analysis Driven Ligand Design for Regioselective Ti-Catalyzed Pyrrole Synthesis.
The rational design of catalysts remains a challenging endeavor within the broader chemical community owing to the myriad variables that can affect key bond-forming events. Designing selective catalysts for any reaction requires an efficient strategy for discovering predictive structure-activity relationships. Herein, we describe the use of iterative supervised principal component analysis (ISPCA) in de novo catalyst design. The regioselective synthesis of 2,5-dimethyl-1,3,4-triphenyl-1H-pyrrole (C) via a Ti-catalyzed formal [2 + 2 +1] cycloaddition of phenylpropyne and azobenzene was targeted as a proof of principle. The initial reaction conditions led to an unselective mixture of all possible pyrrole regioisomers. ISPCA was conducted on a training set of catalysts, and their performance was regressed against the scores from the top three principal components. Component loadings from this PCA space and k-means clustering were used to inform the design of new test catalysts. The selectivity of a prospective test set was predicted in silico using the ISPCA model, and optimal candidates were synthesized and tested experimentally. This data-driven predictive-modeling workflow was iterated, and after only three generations the catalytic selectivity was improved from 0.5 (statistical mixture of products) to over 11 (>90% C) by incorporating 2,6-dimethyl-4-(pyrrolidin-1-yl)pyridine as a ligand. The origin of catalyst selectivity was probed by examining ISPCA variable loadings in combination with DFT modeling, revealing that ligand lability plays an important role in selectivity. A parallel catalyst search using multivariate linear regression (MLR), a popular approach in catalysis informatics, was also conducted in order to compare these strategies in a hypothetical catalyst scouting campaign. ISPCA appears to be more robust and predictive than MLR when sparse training sets are used that are representative of the data available during the early search for an optimal catalyst. The successful development of a highly selective catalyst without resorting to long, stochastic screening processes demonstrates the inherent power of ISPCA in de novo catalyst design and should motivate the general use of ISPCA in reaction development.


Ursodeoxycholic acid inhibits eosinophil degranulation in patients with primary biliary cirrhosis.
Eosinophilia is a distinctive feature of primary biliary cirrhosis (PBC), especially in its early stages. Intriguingly, treatment with ursodeoxycholic acid (UDCA) ameliorates eosinophilia as well as liver tests in patients with PBC. It remains unknown, however, whether eosinophils in PBC patients are functionally activated and whether UDCA inhibits eosinophil activation. In the present study, we systematically examined eosinophil dynamics in the blood and liver in patients with stage I to II PBC before and after UDCA treatment. We determined serum concentrations of eosinophil granule proteins (major basic protein [MBP] and eosinophil-derived neurotoxin [EDN]) by radioimmunoassay and quantitated eosinophil degranulation using computer-assisted morphometry after MBP immunohistochemistry. Before UDCA treatment, patients with PBC (n = 25) showed significantly higher circulating eosinophil counts (P <. 05) and serum concentrations of MBP (P <.0005) and EDN (P <.02) compared with patients with chronic viral hepatitis (n = 22), autoimmune hepatitis (n = 10), and obstructive jaundice (n = 12). Four-week UDCA treatment significantly reduced blood eosinophil counts (P <.0001) and serum MBP (P <.0001) and EDN (P <.0001) levels in PBC patients. MBP immunohistochemistry and computer-assisted quantitative morphometry showed infiltration and degranulation of eosinophils in the portal tract in patients with PBC and significant reductions in the number of sites and the area occupied by extracellular MBP deposits after UDCA treatment for 2 years (P <.02) but not in placebo-treated patients. Our results suggest that eosinophils in patients with PBC are not only increased in number, but also release granule proteins, and that UDCA treatment inhibits this eosinophil activation/degranulation.


The role of intestinal endotoxin in liver injury: a long and evolving history.
From the mid-1950s, it was observed that liver injury by a variety of toxins greatly sensitized the host to the effects of administered lipopolysaccharide. In the nutritional cirrhosis of choline deficiency, and in acute toxic injury as well, the need for the presence of enteric endotoxin was demonstrated. The universality of this association was striking for almost all agents associated with liver injury. In addition, the presence of endotoxemia in human liver disease was documented in the 1970s, when the hypothesis was first proposed, and correlated with the severity of the disease. Despite imposing evidence of the critical role of enteric endotoxin in liver injury, it did not excite much interest in investigators until the 1980s. With the ability to study effects of alcohol in newer delivery systems, and an increased understanding of the role of Kupffer cells in the process, the original hypothesis has been accepted. This historical review details the progress of this novel concept of disease initiation and suggests future directions to bring potential therapies to the bedside.


The coexistence of antiphospholipid syndrome and systemic lupus erythematosus in Colombians.
To examine the prevalence and associated factors related to the coexistence of antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE) in a cohort of Colombian patients with SLE, and to discuss the coexistence of APS with other autoimmune diseases (ADs). A total of 376 patients with SLE were assessed for the presence of the following: 1) confirmed APS; 2) positivity for antiphospholipid (aPL) antibodies without a prior thromboembolic nor obstetric event; and 3) SLE patients without APS nor positivity for aPL antibodies. Comparisons between groups 1 and 3 were evaluated by bivariate and multivariate analysis. Although the prevalence of aPL antibodies was 54%, APS was present in just 9.3% of SLE patients. In our series, besides cardiovascular disease (AOR 3.38, 95% CI 1.11-10.96, p = 0.035), pulmonary involvement (AOR 5.06, 95% CI 1.56-16.74, p = 0.007) and positivity for rheumatoid factor (AOR 4.68, 95%IC 1.63-14.98, p = 0.006) were factors significantly associated with APS-SLE. APS also may coexist with rheumatoid arthritis, Sjögren's syndrome, autoimmune thyroid diseases, systemic sclerosis, systemic vasculitis, dermatopolymyositis, primary biliary cirrhosis and autoimmune hepatitis. APS is a systemic AD that may coexist with other ADs, the most common being SLE. Awareness of this polyautoimmunity should be addressed promptly to establish strategies for controlling modifiable risk factors in those patients.


International Society of Geriatric Oncology consensus on geriatric assessment in older patients with cancer.
To update the International Society of Geriatric Oncology (SIOG) 2005 recommendations on geriatric assessment (GA) in older patients with cancer. SIOG composed a panel with expertise in geriatric oncology to develop consensus statements after literature review of key evidence on the following topics: rationale for performing GA; findings from a GA performed in geriatric oncology patients; ability of GA to predict oncology treatment–related complications; association between GA findings and overall survival (OS); impact of GA findings on oncology treatment decisions; composition of a GA, including domains and tools; and methods for implementing GA in clinical care. GA can be valuable in oncology practice for following reasons: detection of impairment not identified in routine history or physical examination, ability to predict severe treatment-related toxicity, ability to predict OS in a variety of tumors and treatment settings, and ability to influence treatment choice and intensity. The panel recommended that the following domains be evaluated in a GA: functional status, comorbidity, cognition, mental health status, fatigue, social status and support, nutrition, and presence of geriatric syndromes. Although several combinations of tools and various models are available for implementation of GA in oncology practice, the expert panel could not endorse one over another. There is mounting data regarding the utility of GA in oncology practice; however, additional research is needed to continue to strengthen the evidence base.


The distribution of polar ejection forces determines the amplitude of chromosome directional instability.
Polar ejection forces have often been hypothesized to guide directional instability of mitotic chromosomes, but a direct link has never been established. This has led, in part, to the resurgence of alternative theories. By taking advantage of extremely precise femtosecond pulsed laser microsurgery, we abruptly alter the magnitude of polar ejection forces by severing vertebrate chromosome arms. Reduction of polar ejection forces increases the amplitude of directional instability without altering other characteristics, thus establishing a direct link between polar ejection forces and the direction of chromosome movements. We find that polar ejection forces limit the range of chromosome oscillations by increasing the probability that motors at a leading kinetochore abruptly disengage or turn off, leading to a direction reversal. From the relation between the change in oscillation amplitude and the amount a chromosome arm is shortened, we are able to map the distribution of polar ejection forces across the spindle, which is surprisingly different from previously assumed distributions. These results allow us to differentiate between the mechanisms proposed to underlie the directional instability of chromosomes.


Single molecule based SNP detection using designed DNA carriers and solid-state nanopores.
Single nucleotide polymorphisms (SNPs) play a crucial role as molecular markers in medical and diagnostic application. We demonstrate a nanopore based method for SNP detection at the single molecule level. Designed DNA carriers are used to distinguish DNA strands containing only one single base difference and follow strand displacement kinetics.


Basic developmental rules and their implications for epilepsy in the immature brain.
The construction of the human brain with its 10(15) synapses follows a set of complex developmentally and environmentally regulated steps. A series of sequences have been described that are instrumental, in the sense that a failure of any one of them leads to dramatic, life-long consequences. Hence the importance of determining the sequential maturation of neurons, synapses and cortical maps. It is also important to determine how network-driven events become installed, as neuronal activity intervenes in all of these steps and modulates, for better or worse, the outcome. A fundamental consequence of these sequential events is that any disruption will have very different consequences depending on when it occurs, indeed, "when is as important as what". An obvious aspect of these general features is related to seizures. In fact, the developing brain has both a higher incidence of seizures in human and animal models, and experiences seizures that can produce long-lasting consequences that are also stage-dependent. This seminar and the series of slides presented are an introduction to these issues, summing up several studies made notably by INMED researchers during the last two decades (http://www.inmednet.com). It concentrates on four basic developmental rules: i) the generation by very immature neurons, of very large currents mediated by the activation of receptors in neurons that bear no synapses. This is due to the release of GABA that diffuses to distal sites and acts as a paracrine factor; ii) the excitatory/inhibitory shift of the actions of GABA during development because of a progressive reduction in the intracellular chloride concentration; iii) the sequential formation of GABAergic synapses and networks before glutamatergic ones, implying that, at an early stage, all the excitatory drive will be GABAergic; iv) the presence, at an early stage, of a unique, primitive pattern in all developing structures, this pattern disappears when most GABAergic synapses have shifted to their adult configuration. Several consequences of these sequences are described including: i) a control of neuronal migration by GABA-acting drugs, and the possibility that migration disorders are also generated by environmental factors that include the effects of GABA-acting agents; ii) If GABA excites immature neurons and inhibits adult one, then GABA-acting agents will also produce different effects on the mother and the embryo; iii) early brain oscillations are generated by the periphery and propagate centrally - notably to the sensory-motor cortex, suggesting that peripherally-generated movements may provide an important signal for the formation of cortical maps, in keeping with the importance of embryonic movements; iv) "seizures beget seizures" in the developing brain. This has now been shown in a triple chamber with the two intact hippocampi that we developed, and with which it has been possible to show that only recurrent seizures that include high frequency oscillations can transform the naïve, contralateral hippocampus to an epileptic one that seizes spontaneously. Most interestingly, at an early developmental stage, when GABA excites many neurons and the density of glutamatergic synapses is not sufficiently high, purely glutamatergic seizures cannot lead to long-term consequences, the additional excitatory drive provided by GABAergic synapses is needed. In other words, at that stage, blocking GABA synapses generates seizures, as in adults, but these do not lead to long-term consequences. The mechanisms that underlie these differences is due to the need for high frequency oscillations (> 80 Hz or so), and these can only be generated when GABA synapses are operative in the developing brain: GABA receptor antagonists are ictogenic, but not epileptogenic. To facilitate teaching purposes the paper is published together with supplemental data (as a PowerPoint presentation included in the accompanying DVD), thus allowing an overview of important developmental steps and their implications.


The appropriate use of approximate entropy and sample entropy with short data sets.
Approximate entropy (ApEn) and sample entropy (SampEn) are mathematical algorithms created to measure the repeatability or predictability within a time series. Both algorithms are extremely sensitive to their input parameters: m (length of the data segment being compared), r (similarity criterion), and N (length of data). There is no established consensus on parameter selection in short data sets, especially for biological data. Therefore, the purpose of this research was to examine the robustness of these two entropy algorithms by exploring the effect of changing parameter values on short data sets. Data with known theoretical entropy qualities as well as experimental data from both healthy young and older adults was utilized. Our results demonstrate that both ApEn and SampEn are extremely sensitive to parameter choices, especially for very short data sets, N ≤ 200. We suggest using N larger than 200, an m of 2 and examine several r values before selecting your parameters. Extreme caution should be used when choosing parameters for experimental studies with both algorithms. Based on our current findings, it appears that SampEn is more reliable for short data sets. SampEn was less sensitive to changes in data length and demonstrated fewer problems with relative consistency.


European Society of Cardiology guidance for the diagnosis and management of cardiovascular disease during the COVID-19 pandemic: part 1-epidemiology, pathophysiology, and diagnosis.
Since its emergence in early 2020, the novel severe acute respiratory syndrome coronavirus 2 causing coronavirus disease 2019 (COVID-19) has reached pandemic levels, and there have been repeated outbreaks across the globe. The aim of this two-part series is to provide practical knowledge and guidance to aid clinicians in the diagnosis and management of cardiovascular disease (CVD) in association with COVID-19. A narrative literature review of the available evidence has been performed, and the resulting information has been organized into two parts. The first, reported here, focuses on the epidemiology, pathophysiology, and diagnosis of cardiovascular (CV) conditions that may be manifest in patients with COVID-19. The second part, which will follow in a later edition of the journal, addresses the topics of care pathways, treatment, and follow-up of CV conditions in patients with COVID-19. This comprehensive review is not a formal guideline but rather a document that provides a summary of current knowledge and guidance to practicing clinicians managing patients with CVD and COVID-19. The recommendations are mainly the result of observations and personal experience from healthcare providers. Therefore, the information provided here may be subject to change with increasing knowledge, evidence from prospective studies, and changes in the pandemic. Likewise, the guidance provided in the document should not interfere with recommendations provided by local and national healthcare authorities.


PDE4: arrested at the border.
Compartmentation plays a critical role in determining the specificity and efficacy of signaling by adenosine 3',5'-monophosphate (cAMP) and other second messengers. It has become apparent that all of the protein components of the cAMP signaling pathway are subject to regulation and may be localized to particular subcellular compartments. This Perspective discusses recent research concerning the importance of cAMP phosphodiesterase localization in cAMP signaling, functional implications of the organization of this pathway's protein components into macromolecular complexes, and common themes in the compartmentation of cAMP- and Ca2+-mediated signaling pathways.