Bone marrow-derived stromal cells (BMSCs) interact with fibroblasts in accelerating wound healing.
Bone marrow-derived stromal cells (BMSCs) exhibit extraordinary degree of plasticity and growth factor repertoire for which they have been investigated for repair and regeneration of damaged tissues, but have not been adequately examined for wound healing. The ability of BMSCs to accelerate healing of surgically inflicted cutaneous and fascial wounds was tested in vivo in rats and in vitro using a fibroblast monolayer wound model. Intravenous treatment with BMSCs augmented healing of both cutaneous and fascial wounds as determined by an increase in the biomechanical strength of wounds. In vitro experiments showed that incorporation of BMSCs in fibroblast monolayers accelerates the closure of mechanically disrupted monolayers, which was attributed to the enhanced migration of fibroblasts onto the denuded surfaces. Furthermore, culture medium conditioned by activated BMSCs promoted the closure of defects in monolayers and enhanced the proliferation/growth and directional migration (chemotaxis) of fibroblasts. This study demonstrates that BMSCs significantly augment healing of cutaneous and fascial wounds in vivo at least in part through interaction with fibroblasts in which BMSCs promote growth and chemotaxis of fibroblasts.


Quantifying the roles of host movement and vector dispersal in the transmission of vector-borne diseases of livestock.
The role of host movement in the spread of vector-borne diseases of livestock has been little studied. Here we develop a mathematical framework that allows us to disentangle and quantify the roles of vector dispersal and livestock movement in transmission between farms. We apply this framework to outbreaks of bluetongue virus (BTV) and Schmallenberg virus (SBV) in Great Britain, both of which are spread by Culicoides biting midges and have recently emerged in northern Europe. For BTV we estimate parameters by fitting the model to outbreak data using approximate Bayesian computation, while for SBV we use previously derived estimates. We find that around 90% of transmission of BTV between farms is a result of vector dispersal, while for SBV this proportion is 98%. This difference is a consequence of higher vector competence and shorter duration of viraemia for SBV compared with BTV. For both viruses we estimate that the mean number of secondary infections per infected farm is greater than one for vector dispersal, but below one for livestock movements. Although livestock movements account for a small proportion of transmission and cannot sustain an outbreak on their own, they play an important role in establishing new foci of infection. However, the impact of restricting livestock movements on the spread of both viruses depends critically on assumptions made about the distances over which vector dispersal occurs. If vector dispersal occurs primarily at a local scale (99% of transmission occurs <25 km), movement restrictions are predicted to be effective at reducing spread, but if dispersal occurs frequently over longer distances (99% of transmission occurs <50 km) they are not.


Parkinson's disease dementia and potential therapeutic strategies.
Dementia in Parkinson's disease (PD-D) has only been acknowledged in the recent three decades, but research in this field has accelerated. The purpose of this review was to discuss advances in PD-D regarding biomarker correlates and potential therapeutic targets. Attention and executive dysfunction, memory deficits that improve with cueing, and visual hallucinations are characteristic in PD-D. PD-D dramatically increases the disability and misery of the disease. Current treatment for PD-D is symptomatic, modest, and only transiently effective. There is wide agreement that more effective treatment is needed, but this will require more knowledge about PD-D pathophysiology. Advances in the pathogenesis of PD have focused on the substantia nigra, which is the location from where the pathophysiology of motor symptoms primarily arises in initial stages. In contradistinction, pathology studies have suggested that cognitive decline correlates with cortical and subcortical-cortical projection pathway abnormalities. There is evidence that substantia nigra mechanisms, including protein aggregation of α-synuclein (e.g., Lewy bodies) may also play a role in cortical neuron degeneration. Other different mechanisms, such as Alzheimer's disease pathology (e.g., Aβ aggregation) may be operant for PD-D. Biomarkers of various types are being proposed for the study of PD-D as well as for objective measures of PD-D prediction and progression. Therapeutic targets are currently derived mostly from general PD neurodegeneration research rather than cortical PD neurodegeneration per se. Protein aggregation, genes that are associated with PD, oxidative stress, inflammation, and trophic factors constitute the major classes of therapeutic targets for PD-D. More research is needed on the specific aspects of cortical dysfunction and degeneration that create PD-D.


Theory of concentric circular grid.
Any image-forming optical system is usually tested for aberrations by looking at the image plane. A point source is placed at some distance from the system (this may be a very large or small distance depending on the particular system) and its image by the optical system is studied by any of many methods available. The present paper presents the theory of a method in which a grating containing a series of concentric circles is placed at the image of the point source. Characteristic patterns are derived for the usual aberrations of optical systems and some typical photographs of these patterns are presented. Finally, this method is compared to the Ronchi test in which a series of straight lines are used for the grating.


Biological support media influence the bacterial biofouling community in reverse osmosis water reclamation demonstration plants.
The diversity of the bacterial community developed in different stages of two reverse osmosis (RO) water reclamation demonstration plants designed in a wastewater treatment plant (WWTP) in Tarragona (Spain) was characterized by applying 454-pyrosequencing of the 16S rRNA gene. The plants were fed by secondary treated effluent to a conventional pretreatment train prior to the two-pass RO system. Plants differed in the material used in the filtration process, which was sand in one demonstration plant and Scandinavian schists in the second plant. The results showed the presence of a highly diverse and complex community in the biofilms, mainly composed of members of the Betaproteobacteria and Bacteroidetes in all stages, with the presence of some typical wastewater bacteria, suggesting a feed water origin. Community similarities analyses revealed that samples clustered according to filter type, highlighting the critical influence of the biological supporting medium in biofilm community structure.


Increased circulating levels of betatrophin in individuals with long-standing type 1 diabetes.
The hormone betatrophin was recently described as a potent stimulator of beta cell proliferation in mice. Insulin resistance, but not insulin deficiency, caused upregulation of betatrophin expression. If these findings were found to be fully applicable in humans, this would open up the possibility of future betatrophin treatment in type 1 diabetes. The present study measured for the first time betatrophin concentrations in humans and tested the hypothesis that there would be no difference in circulating betatrophin concentrations between patients with type 1 diabetes and healthy individuals. Betatrophin concentrations in plasma of 33 patients with type 1 diabetes and 24 age-matched healthy controls were measured by ELISA. The study participants were characterised for blood lipids, BMI, plasma glucose and HbA1c, and, for the diabetic patients, their insulin requirements and any residual C-peptide concentrations. Plasma betatrophin concentrations were normally ~300 pg/ml, but were approximately doubled in patients with type 1 diabetes. In the patients, there were no correlations between betatrophin and age, blood lipids, BMI, glucose control or insulin requirement, whereas in controls betatrophin levels increased with age. BMI, blood pressure and triacylglycerol, LDL-cholesterol and HDL-cholesterol levels were similar in patients and healthy controls. Circulating concentrations of betatrophin are increased in type 1 diabetes in contrast with what was recently described in an insulin-deficient mouse model. However, increased betatrophin concentrations do not protect against loss of C-peptide. Betatrophin treatment in type 1 diabetes would therefore probably not be successful without the use of supraphysiological doses or a combination with immune regulatory treatment.


Determination of retardation parameters of multiple-order wave plate using a phase-sensitive heterodyne ellipsometer.
To characterize the linear birefringence of a multiple-order wave plate (MWP), an oblique incidence is one of the methods available. Multiple reflections in the MWP are produced, and oscillations in the phase retardation measurement versus the oblique incident angle are then measured. Therefore, an antireflection coated MWP is required to avoid oscillation of the phase retardation measurement. In this study, we set up a phase-sensitive heterodyne ellipsometer to measure the phase retardations of an uncoated MWP versus the oblique incident angle, which was scanned in the x-z plane and y-z plane independently. Thus, the effect on multiple reflections by the MWP is reduced by means of subtracting the two measured phase retardations from each other. As a result, a highly sensitive and accurate measurement of retardation parameters (RPs), which includes the refractive indices of the extraordinary ray n(e) and ordinary ray n(o), is obtained by this method. On measurement, a sensitivity (n(e),n(o)) of 10(-6) was achieved by this experiment setup. At the same time, the spatial shifting of the P and S waves emerging from the MWP introduced a deviation between experimental results and the theoretical calculation.


Induced biliary excretion of Listeria monocytogenes.
Listeria monocytogenes is a ubiquitous gram-positive bacterium that can cause systemic and often life-threatening disease in immunocompromised hosts. This organism is largely an intracellular pathogen; however, we have determined that it can also grow extracellularly in animals, in the lumen of the gallbladder. The significance of growth in the gallbladder with respect to the pathogenesis and spread of listeriosis depends on the ability of the bacterium to leave this organ and be disseminated to other tissues and into the environment. Should this process be highly inefficient, growth in the gallbladder would have no impact on pathogenesis or spread, but if it occurs efficiently, bacterial growth in this organ may contribute to listeriosis and dissemination of this organism. Here, we use whole-body imaging to determine the efficacy and kinetics of food- and hormone-induced biliary excretion of L. monocytogenes from the murine gallbladder, demonstrating that transit through the bile duct into the intestine can occur within 5 min of induction of gallbladder contraction by food or cholecystokinin and that movement of bacteria through the intestinal lumen can occur very rapidly in the absence of fecal material. These studies demonstrate that L. monocytogenes bacteria replicating in the gallbladder can be expelled from the organ efficiently and that the released bacteria move into the intestinal tract, where they pass into the environment and may possibly reinfect the animal.


Protons and Hydroxide Ions in Aqueous Systems.
Understanding the structure and dynamics of water's constituent ions, proton and hydroxide, has been a subject of numerous experimental and theoretical studies over the last century. Besides their obvious importance in acid-base chemistry, these ions play an important role in numerous applications ranging from enzyme catalysis to environmental chemistry. Despite a long history of research, many fundamental issues regarding their properties continue to be an active area of research. Here, we provide a review of the experimental and theoretical advances made in the last several decades in understanding the structure, dynamics, and transport of the proton and hydroxide ions in different aqueous environments, ranging from water clusters to the bulk liquid and its interfaces with hydrophobic surfaces. The propensity of these ions to accumulate at hydrophobic surfaces has been a subject of intense debate, and we highlight the open issues and challenges in this area. Biological applications reviewed include proton transport along the hydration layer of various membranes and through channel proteins, problems that are at the core of cellular bioenergetics.


Characteristics of ultra-long range surface plasmon waves at optical frequencies.
It has been reported earlier that ultra-long range surface plasmon waves can be supported if dielectric layers with lower index of refraction than that of the dielectric cladding are placed on either side of the thin metal film. In this paper, we report a further investigation of the ultra-long range surface plasmon modes and the condition to support such ultra-long propagation distances at optical frequencies. We studied the effects of the index of refraction contrast between the inner layer and the cladding dielectrics, the metal film thickness, and the dispersion with wavelength. We present a condition which must be satisfied by the waveguide structure to support the bound ultra-long range surface plasmon mode.


Chemoselective Ullmann coupling at room temperature: a facile access to 2-aminobenzo[b]thiophenes.
Various functionalized 2-aminobenzo[b]thiophenes have been synthesized at room temperature by the Ullmann coupling reaction for the first time. The enantiospecific coupling reaction has been further demonstrated without loss of optical purity. The newly synthesized 2-anilino-3-cyano-benzo[b]thiophenes are transformed into 11-amino-benzothieno[2,3-b]quinolines in the presence of triflic acid.


Fast global image smoothing based on weighted least squares.
This paper presents an efficient technique for performing a spatially inhomogeneous edge-preserving image smoothing, called fast global smoother. Focusing on sparse Laplacian matrices consisting of a data term and a prior term (typically defined using four or eight neighbors for 2D image), our approach efficiently solves such global objective functions. In particular, we approximate the solution of the memory-and computation-intensive large linear system, defined over a d-dimensional spatial domain, by solving a sequence of 1D subsystems. Our separable implementation enables applying a linear-time tridiagonal matrix algorithm to solve d three-point Laplacian matrices iteratively. Our approach combines the best of two paradigms, i.e., efficient edge-preserving filters and optimization-based smoothing. Our method has a comparable runtime to the fast edge-preserving filters, but its global optimization formulation overcomes many limitations of the local filtering approaches. Our method also achieves high-quality results as the state-of-the-art optimization-based techniques, but runs ∼10-30 times faster. Besides, considering the flexibility in defining an objective function, we further propose generalized fast algorithms that perform Lγ norm smoothing (0 < γ < 2) and support an aggregated (robust) data term for handling imprecise data constraints. We demonstrate the effectiveness and efficiency of our techniques in a range of image processing and computer graphics applications.


Preterm heart in adult life: cardiovascular magnetic resonance reveals distinct differences in left ventricular mass, geometry, and function.
Preterm birth leads to an early switch from fetal to postnatal circulation before completion of left ventricular in utero development. In animal studies, this results in an adversely remodeled left ventricle. We determined whether preterm birth is associated with a distinct left ventricular structure and function in humans. A total of 234 individuals 20 to 39 years of age underwent cardiovascular magnetic resonance. One hundred two had been followed prospectively since preterm birth (gestational age=30.3±2.5 week; birth weight=1.3±0.3 kg), and 132 were born at term to uncomplicated pregnancies. Longitudinal and short-axis cine images were used to quantify left ventricular mass, 3-dimensional geometric variation by creation of a unique computational cardiac atlas, and myocardial function. We then determined whether perinatal factors modify these left ventricular parameters. Individuals born preterm had increased left ventricular mass (66.5±10.9 versus 55.4±11.4 g/m(2); P<0.001) with greater prematurity associated with greater mass (r = -0.22, P=0.03). Preterm-born individuals had short left ventricles with small internal diameters and a displaced apex. Ejection fraction was preserved (P>0.99), but both longitudinal systolic (peak strain, strain rate, and velocity, P<0.001) and diastolic (peak strain rate and velocity, P<0.001) function and rotational (apical and basal peak systolic rotation rate, P =0.05 and P =0.006; net twist angle, P=0.02) movement were significantly reduced. A diagnosis of preeclampsia during the pregnancy was associated with further reductions in longitudinal peak systolic strain in the offspring (P=0.02, n=29). Individuals born preterm have increased left ventricular mass in adult life. Furthermore, they exhibit a unique 3-dimensional left ventricular geometry and significant reductions in systolic and diastolic functional parameters. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01487824.


Comparison of Leishmania typing results obtained from 16 European clinical laboratories in 2014.
Leishmaniasis is endemic in southern Europe, and in other European countries cases are diagnosed in travellers who have visited affected areas both within the continent and beyond. Prompt and accurate diagnosis poses a challenge in clinical practice in Europe. Different methods exist for identification of the infecting Leishmania species. Sixteen clinical laboratories in 10 European countries, plus Israel and Turkey, conducted a study to assess their genotyping performance. DNA from 21 promastigote cultures of 13 species was analysed blindly by the routinely used typing method. Five different molecular targets were used, which were analysed with PCR-based methods. Different levels of identification were achieved, and either the Leishmania subgenus, species complex, or actual species were reported. The overall error rate of strains placed in the wrong complex or species was 8.5%. Various reasons for incorrect typing were identified. The study shows there is considerable room for improvement and standardisation of Leishmania typing. The use of well validated standard operating procedures is recommended, covering testing, interpretation, and reporting guidelines. Application of the internal transcribed spacer 1 of the rDNA array should be restricted to Old World samples, while the heat-shock protein 70 gene and the mini-exon can be applied globally.


Hybridization of Evolutionary Operators with Elitist Iterated Racing for the Simulation Optimization of Traffic Lights Programs.
In the traffic light scheduling problem, the evaluation of candidate solutions requires the simulation of a process under various (traffic) scenarios. Thus, good solutions should not only achieve good objective function values, but they must be robust (low variance) across all different scenarios. Previous work has shown that combining IRACE with evolutionary operators is effective for this task due to the power of evolutionary operators in numerical optimization. In this article, we further explore the hybridization of evolutionary operators and the elitist iterated racing of IRACE for the simulation-optimization of traffic light programs. We review previous works from the literature to find the evolutionary operators performing the best when facing this problem to propose new hybrid algorithms. We evaluate our approach over a realistic case study derived from the traffic network of Málaga (Spain) with 275 traffic lights that should be scheduled optimally. The experimental analysis reveals that the hybrid algorithm comprising IRACE plus differential evolution offers statistically better results than the other algorithms when the budget of simulations is low. In contrast, IRACE performs better than the hybrids for a high simulations budget, although the optimization time is much longer.


In the shadow of Apartheid: intergenerational transmission of Black parental trauma as it emerges in the analytical space of inter-racial subjectivities.
Using the construct of projective identification and integrating it with the body of literature on intergenerational transmission of unsymbolized parental trauma, I describe the case of an adult black South African woman called Sibulelo. It is suggested that Sibulelo has unconsciously identified with the disavowed parents and grandparents trauma that they suffered as a result of the system of Apartheid. Such trauma is expressed through her feelings of being dis-located in time and space, as if she is living outside of herself, unplugged from life, and living someone else's life. The paper details the unfolding therapeutic process in relation to my whiteness in the context of her blackness. This brings into sharp focus an exploration of black-white racialized transference-counter-transference matrix in the context of intergenerational trauma. It is a reflective paper and opens up my own counter-transference, thus foregrounding the notion of therapeutic inter-subjectivity. A further contribution to psychoanalytic theory concerns the role of recognition and being seen as a powerful process in facilitating the symbolization of trauma. In addition, if there is no interruption of the cycles of intergenerational trauma, and therefore no symbolization, it becomes an unconscious familial compulsion to repeat. Moreover, this therapy case highlights the idea that as a traumatised family living within a bruised culture of intergenerational transmission of trauma, such repetition of trauma becomes a cultural compulsion to repeat what has not been spoken or named.


Off-label prescribing in palliative care - a cross-sectional national survey of palliative medicine doctors.
Regulatory bodies including the European Medicines Agency register medications (formulation, route of administration) for specific clinical indications. Once registered, prescription is at clinicians' discretion. Off-label use is beyond the registered use. While off-label prescribing may, at times, be appropriate, efficacy and toxicity data are often lacking. The aim of this study was to document off-label use policies (including disclosure and consent) in Australian palliative care units and current practices by palliative care clinicians. A national, cross-sectional survey was conducted online following an invitation letter. The survey asked clinicians their most frequent off-label medication/indication dyads and unit policies. Dyads were classified into unregistered, off-label and on-label, and for the latter, whether medications were nationally subsidised. All Australian palliative medicine Fellows and advanced trainees. Overall, 105 clinicians responded (53% response rate). The majority did not have policies on off-label medications, and documented consent rarely. In all, 236 medication/indication dyads for 36 medications were noted: 45 dyads (19%) were for two unregistered medications, 118 dyads (50%) were for 26 off-label medications and 73 dyads (31%) were for 12 on-label medications. Off-label prescribing with its clinical, legal and ethical implications is common yet poorly recognised by clinicians. A distinction needs to be made between where quality evidence exists but registration has not been updated by the pharmaceutical sponsor and the evidence has not been generated. Further research is required to quantify any iatrogenic harm from off-label prescribing in palliative care.


Anionic halocuprate(II) complexes as catalysts for the oxaziridine-mediated aminohydroxylation of olefins.
We have discovered that the oxaziridine-mediated copper-catalyzed aminohydroxylation reaction recently discovered in our laboratories is dramatically accelerated in the presence of halide additives. The use of this more active catalyst system enables the efficient aminohydroxylation of electronically and sterically deactivated styrenes and also enables the use of nonstereogenic 3,3-dialkyl oxaziridines as terminal oxidants in the aminohydroxylation reaction. We present evidence that anionic halocuprate(II) complexes are the catalytically active species responsible for the increased reactivity under these conditions. This unexpected observation has led us to re-evaluate our mechanistic understanding of this reaction. On the basis of the results of a variety of radical trapping experiments, we propose a modified mechanism that involves a homolytic reaction of the olefin with a copper(II)-activated oxaziridine. Together, the observation that anionic additives significantly increase the oxidizing ability of oxaziridines and the recognition of the radical nature of reactions of oxaziridines under these conditions suggest that a variety of new oxidative transformations catalyzed by halocuprate(II) complexes should be possible.


Secondhand Smoke Exposure in Childhood and Adulthood in Relation to Adult Mortality Among Never Smokers.
Secondhand smoke is known to have adverse effects on the lung and vascular systems in both children and adults. It is unknown if childhood exposure to secondhand smoke is associated with adult mortality. The authors examined associations of childhood and adult secondhand smoke exposure with death from all causes, ischemic heart disease, stroke, and chronic obstructive pulmonary disease among 70,900 never smoking men and women, predominantly aged ≥50 years, from the Cancer Prevention Study-II Nutrition Cohort in 1992-1993. There were 25,899 participant deaths during follow-up through 2014. During 2016-2017, Cox proportional hazards regression models were used to calculate multivariable-adjusted hazard ratios and 95% CIs. Childhood secondhand smoke exposure was not associated with all-cause mortality. However, childhood secondhand smoke (living with a smoker for 16-18 years during childhood) was associated with higher mortality from chronic obstructive pulmonary disease (hazard ratio=1.31, 95% CI=1.05, 1.65). Adult secondhand smoke exposure of ≥10 hours/week at enrollment was associated with a higher risk of all-cause (hazard ratio=1.09, 95% CI=1.04, 1.14); ischemic heart disease (hazard ratio=1.27, 95% CI=1.14, 1.42); stroke (hazard ratio=1.23, 95% CI=1.04, 1.45); and chronic obstructive pulmonary disease (hazard ratio=1.42, 95% CI=0.97, 2.09) mortality. These results suggest that childhood secondhand smoke exposure, as well as adult secondhand smoke exposure, increase the risk of chronic obstructive pulmonary disease death in adulthood. Consistent with previous studies, the results also show that adult secondhand smoke is meaningfully associated with higher mortality from vascular disease and all causes. Overall, these findings provide further evidence for reducing secondhand smoke exposure throughout life.


Osteoblast-specific expression of the fibrous dysplasia (FD)-causing mutation Gsα(R201C) produces a high bone mass phenotype but does not reproduce FD in the mouse.
We recently reported the generation and initial characterization of the first direct model of human fibrous dysplasia (FD; OMIM #174800), obtained through the constitutive systemic expression of one of the disease-causing mutations, Gsα(R201C) , in the mouse. To define the specific pathogenetic role(s) of individual cell types within the stromal/osteogenic system in FD, we generated mice expressing Gsα(R201C) selectively in mature osteoblasts using the 2.3kb Col1a1 promoter. We show here that this results in a striking high bone mass phenotype but not in a mimicry of human FD. The high bone mass phenotype involves specifically a deforming excess of cortical bone and prolonged and ectopic cortical bone remodeling. Expression of genes characteristic of late stages of bone cell differentiation/maturation is profoundly altered as a result of expression of Gsα(R201C) in osteoblasts, and expression of the Wnt inhibitor Sost is reduced. Although high bone mass is, in fact, a feature of some types/stages of FD lesions in humans, it is marrow fibrosis, localized loss of adipocytes and hematopoietic tissue, osteomalacia, and osteolytic changes that together represent the characteristic pathological profile of FD, as well as the sources of specific morbidity. None of these features are reproduced in mice with osteoblast-specific expression of Gsα(R201C) . We further show that hematopoietic progenitor/stem cells, as well as more mature cell compartments, and adipocyte development are normal in these mice. These data demonstrate that effects of Gsα mutations underpinning FD-defining tissue changes and morbidity do not reflect the effects of the mutations on osteoblasts proper.


Key treatment questions in childhood acute lymphoblastic leukemia: results in 5 consecutive trials performed by the ALL-BFM study group from 1981 to 2000.
Between 1981 and 2000, 6 609 children (<18 years of age) were treated in 5 consecutive trials of the Berlin-Frankfurt-Münster (BFM) study group for childhood acute lymphoblastic leukemia (ALL). Patients were treated in up to 82 centers in Germany, Austria, and Switzerland. Probability of 10-year event-free survival (survival) improved from 65% (77%) in study ALL-BFM 81-78% (85%) in ALL-BFM 95. In parallel to relapse reduction, major efforts focused on reducing acute and late toxicity through advanced risk adaptation of treatment. The major findings derived from these ALL-BFM trials were as follows: 1) preventive cranial radiotherapy could be safely reduced to 12 Gy in T-ALL and high-risk ALL patients and eliminated in non-high-risk non-T-ALL patients, if it was replaced by high-dose and intrathecal methotrexate; 2) omission of delayed reintensification severely impaired outcome of low-risk patients; 3) 6 months less maintenance therapy caused an increase in systemic relapses; 4) slow response to an initial 7-day prednisone window was identified as adverse prognostic factor; 5) condensed induction therapy resulted in a significant improvement of outcome; 6) the daunorubicin dose in induction could be safely reduced in low-risk patients; 7) intensification of consolidation/reintensification treatment led to considerable improvement of outcome in high-risk patients.


Determination of vanadium as 4-(2-pyridylazo)resorcinol-hydrogen peroxide ternary complexes by ion-interaction reversed-phase liquid chromatography.
The separation and determination of the vanadium(V) ternary complex formed with 4-(2-pyridylazo)resorcinol (PAR) and hydrogen peroxide using ion-interaction reversed-phase high-performance liquid chromatography on a C18 column has been investigated. The optimal mobile phase was a methanol-water solution (32:68, v/v) containing 3 mM tetrabutylammonium bromide, 5 mM acetic acid and 5 mM citrate buffer at pH 7, with absorbance detection at 540 nm. The stoichiometry of the ternary complex of vanadium at pH 6 in 10 mM acetate buffer using the mole ratio and Job's method by HPLC indicated that the mole ratio of V(V):PAR:H2O2 was 1:1:1. The optimal conditions for precolumn formation of the ternary complex were 10 mM acetate, 7 mM H2O2, 0.3 mM PAR, and pH 6. The method gave relative standard deviations of retention time, peak area and peak height for the ternary complex of 0.187, 0.45 and 0.57%, respectively. The detection limit (at a signal-to-noise ratio of 3) for V(V) was 0.09 ng/ml in the digested sample using a 100-microl injection loop (or 0.09 microg/g in the solid fertiliser sample). The method was applied to the analysis of fertilisers (phosphate rocks and nitrogen, phosphorus and potassium fertiliser). The results for vanadium obtained by the HPLC method agreed well with those from magnetic sector inductively coupled plasma MS analysis.


Endoscopic surgery for malignant tumors of the sinonasal tract and adjacent skull base: a 10-year experience.
The increasing expertise in the field of transnasal endoscopic surgery recently has expanded its indications to include the management of sinonasal malignancies. We report our experience with the endoscopic management of nasoethmoidal malignancies possibly involving the adjacent skull base. A retrospective analysis was performed of patients treated by an exclusive endoscopic approach (EEA) or a cranioendoscopic approach (CEA) from 1996 to 2006 managed by two surgical teams at the Departments of Otorhinolaryngology of the University of Brescia, and the University of Pavia/Insubria-Varese, Italy. One-hundred eighty-four patients were considered eligible for the present analysis. An EEA was performed in 134 patients and the remaining 50 patients underwent the CEA. The most frequent histotypes encountered were adenocarcinoma (37%), squamous cell carcinoma (13.6%), olfactory neuroblastoma (12%), mucosal melanoma (9.2%), and adenoid cystic carcinoma (7.1%). Overall, 86 (46.7%) patients received some form of adjuvant treatment. The patients were followed up for a mean of 34.1 months (range, 2-123 months). The 5-year disease-specific survival was 91.4 +/- 3.9% and 58.8 +/- 8.6% (p = 0.0004) for the EEA and CEA group, respectively. To the best of our knowledge, this is the largest series reported to date of malignant tumors of the sinonasal tract and adjacent skull base treated with pure endoscopic or cranioendoscopic techniques. A 5-year disease-specific survival of 91.4% and 58.8% for the EEA and the CEA groups, respectively, seem to indicate that endoscopic surgery, when properly planned and in expert hands, may be a valid alternative to standard surgical approaches for the management of malignancies of the sinonasal tract.


In-vivo myocardial substrate alteration during perfused ventricular fibrillation.
Earlier work suggests the in-vivo heart alters its substrate utilization as a function of cardiac work. Previous work has also demonstrated the high oxygen requirements of the heart during ventricular fibrillation (VF). The authors hypothesized that myocardial substrate utilization during VF with perfusion is similar to the normal beating heart under conditions of increased workload. Myocardial substrate selection was studied in the in-vivo porcine myocardium using 13carbon nuclear magnetic resonance (13C NMR) under conditions of increased cardiac work (dobutamine group) and VF with extracorporeal perfusion (VF group). Once the animal preparation was completed, metabolic steady state was achieved with the infusion of unlabeled acetate into the left anterior descending (LAD) coronary artery. The infused substrate was then changed to [2-13C] acetate and glutamate pool labeling was monitored by 13C NMR. The glutamate C4 resonance areas at baseline and after intervention of either increased workload (dobutamine group) or perfused VF (VF group) were compared within groups using paired t-tests. Baseline aortic and great cardiac vein lactates, glucose levels, blood gases, hemoglobin levels, and temperatures were similar between groups. In both groups, there was a significant decrease from baseline in the labeling of C4 glutamate peaks (dobutamine group: 20.2+/-14.9 vs 84.7+/-32.7, p = 0.002; and VF group: 49.8+/-24.4 vs 83.9+/-24.4, p = 0.02), indicating selection against acetate oxidation in favor of other endogenous substrates. In the in-vivo heart, despite the absence of functional contractions, changes in substrate utilization during perfused VF are similar to changes that occur with increased workload in the normal beating heart.


Coupling between codon usage, translation and protein export in Escherichia coli.
Proteins destined for export via the Sec-dependent pathway are synthesized with a short N-terminal signal peptide. A requirement for export is that the proteins are in a translocationally competent state. This is a loosely folded state that allows the protein to pass through the SecYEG apparatus and pass into the periplasm. In order to maintain pre-secretory proteins in an export-competent state, there are many factors that slow the folding of the pre-secretory protein in the cytoplasm. These include cytoplasmic chaperones, such as SecB, and the signal recognition particle, which bind the pre-secretory protein and direct it to the cytoplasmic membrane for export. Recently, evidence has been published that non-optimal codons in the signal sequence are important for a time-critical early event to allow the correct folding of pre-secretory proteins. This review details the recent developments in folding of the signal peptide and the pre-secretory protein.


Frankincense essential oil suppresses melanoma cancer through down regulation of Bcl-2/Bax cascade signaling and ameliorates heptotoxicity via phase I and II drug metabolizing enzymes.
Melanoma is a deadly form of malignancy and according to the World Health Organization 132,000 new cases of melanoma are diagnosed worldwide each year. Surgical resection and chemo/drug treatments opted for early and late stage of melanoma respectively, however detrimental post surgical and chemotherapy consequences are inevitable. Noticeably melanoma drug treatments are associated with liver injuries such as hepatitis and cholestasis which are very common. Alleviation of these clinical manifestations with better treatment options would enhance prognosis status and patients survival. Natural products which induce cytotoxicity with minimum side effects are of interest to achieve high therapeutic efficiency. In this study we investigated anti-melanoma and hepatoprotective activities of frankincense essential oil (FEO) in both in vitro and in vivo models. Pretreatment with FEO induce a significant (p < 0.05) dose-dependent reduction in the cell viability of mouse (B16-F10) and human melanoma (FM94) but not in the normal human epithelial melanocytes (HNEM). Immunoblot analysis showed that FEO induces down regulation of Bcl-2 and up regulation of BAX in B16-F10 cells whereas in FM94 cells FEO induced dose-dependent cleavage of caspase 3, caspase 9 and PARP. Furthermore, FEO (10 μg/ml) treatment down regulated MCL1 in a time-dependent manner in FM94 cells. In vivo toxicity analysis reveals that weekly single dose of FEO (1200 mg/kg body weight) did not elicit detrimental effect on body weight during four weeks of experimental period. Histology of tissue sections also indicated that there were no observable histopathologic differences in the brain, heart, liver, and kidney compare to control groups. FEO (300 and 600 mg/kg body weight) treatments significantly reduced the tumor burden in C57BL/6 mice melanoma model. Acetaminophen (750 mg/kg body weight) was used to induce hepatic injury in Swiss albino mice. Pre treatment with FEO (250 and 500 mg/kg body weight) for seven days retained hematology (complete blood count), biochemical parameters (AST, ALT, ALK, total bilirubin, total protein, glucose, albumin/globulin ratio, cholesterol and triglyceride), and the level of phase I and II drug metabolizing enzymes (cytochrome P450, cytochromeb5, glutathione-S-transferase) which were obstructed by the administration of acetaminophen. Further liver histology showed that FEO treatments reversed the damages (central vein dilation, hemorrhage, and nuclei condensation) caused by acetaminophen. In conclusion, FEO elicited marked anti-melanoma in both in vitro and in vivo with a significant heptoprotection.


The Bacterial Flagellar Rotary Motor in Action.
The bacterial flagellar motor is one of the few rotary motors in nature. Only ∼50 nm in diameter, this transmembrane, ion-driven nanomachine rotates a semirigid helical flagellum at speeds of up to 1300 rps. It is composed of at least 13 different proteins, in different copy numbers, resulting from the coordinated, sequential expression of more than 40 genes. Structural studies have revealed a great deal of information about the structure of the motor, but the in vivo activity has been more elusive. Using a multidisciplinary approach combining molecular biology with single molecule fluorescence microscopy and novel data analysis recent work has obtained quantitative data on the stoichiometry, dynamics, and turnover of components of functioning motors in vivo under physiological conditions. This has shown that it is not a stable rotary machine, but that its structure is highly dynamic and undergoes adaptive remodeling in response to different intracellular and extracellular signals.


Ketamine augmentation of electroconvulsive therapy to improve neuropsychological and clinical outcomes in depression (Ketamine-ECT): a multicentre, double-blind, randomised, parallel-group, superiority trial.
The use of electroconvulsive therapy (ECT) is limited by concerns about its cognitive adverse effects. Preliminary evidence suggests that administering the glutamate antagonist ketamine with ECT might alleviate cognitive adverse effects and accelerate symptomatic improvement; we tested this in a randomised trial of low-dose ketamine. In this multicentre, randomised, parallel-group study in 11 ECT suites serving inpatient and outpatient care settings in seven National Health Service trusts in the North of England, we recruited severely depressed patients, who were diagnosed as having unipolar or bipolar depressive episodes defined as moderate or severe by DSM-IV criteria, aged at least 18 years, and were able and willing to provide written consent to participate in the study. Patients were randomly assigned (1:1) to ketamine (0·5 mg/kg intravenous bolus) or saline adjunctive to the anaesthetic for the duration of their ECT course. Patients and assessment and ECT treatment teams were masked to treatment allocation, although anaesthetists administering the study medication were not. We analysed the primary outcome, Hopkins Verbal Learning Test-Revised delayed verbal recall (HVLT-R-DR) after four ECT treatments, using a Gaussian repeated measures model in all patients receiving the first ECT treatment. In the same population, safety was assessed by adverse effect monitoring. This trial was registered with International Standard Randomised Controlled Trial Number, number ISRCTN14689382. Between early December, 2012, and mid-June, 2015, 628 patients were screened for eligibility, of whom 79 were randomly assigned to treatment (40 in the ketamine group vs 39 in the saline group). Ketamine (mean 5·17, SD 2·92), when compared with saline (5·54, 3·42), had no benefit on the primary outcome (HVLT-R-DR; difference in means -0·43 [95% CI -1·73 to 0·87]). 15 (45%) of 33 ketamine-treated patients compared with 10 (27%) of 37 patients receiving saline experienced at least one adverse event which included two (6%) of 33 patients who had ketamine-attributable transient psychological effects. Psychiatric adverse events were the most common in both groups (six [27%] of 22 adverse events in the ketamine group vs seven [54%] of 13 in the saline group). No evidence of benefit for ketamine was found although the sample size used was small; however, the results excluded greater than a small to moderate benefit with 95% confidence. The results do not support the use of adjunctive low-dose ketamine in routine ECT treatment. National Institute for Health Research (NIHR) Efficacy and Mechanism Evaluation (EME) programme, an MRC and NIHR partnership.


Sex differences in melanoma survival are not related to mitotic rate of the primary tumor.
Based on prior studies, we concluded that the female advantage in melanoma survival is caused by biological factors and not by differences in patient behavior. In this study, we investigated whether this biological advantage was caused by more aggressive tumors in males, as measured by mitotic rate (MR). Data for patients with complete information on MR, Breslow thickness, ulceration and primary tumor location were extracted from the database of Melanoma Institute Australia in Sydney. A negative binomial regression model was used to assess the independent predictive value of sex for MR. Also, the impact of MR on the sex survival advantage was investigated using Cox proportional hazards models. A total of 9,306 patients were included in the analysis. Although males had a slightly higher MR at diagnosis, sex was not an independent predictor of MR after adjustment for all other prognostic factors: incidence rate ratio 0.98, 95 % confidence interval (CI) 0.93-1.02, p = 0.32. After adjustment for all prognostic factors, females had a survival advantage of 36 % (hazard ratio 0.65, 95 % CI 0.55-0.75, p < 0.001). When added as a confounder, MR did not influence this sex hazard ratio. Sex did not independently predict the aggressiveness of a primary melanoma. Furthermore, MR did not influence the known female survival advantage. Based on these results, the biological trait underlying sex survival differences in melanoma seems not to be tumor-related and therefore is more likely to be caused by host factors.


Estrogen receptor α or β loss in the colon of Min/+ mice promotes crypt expansion and impairs TGFβ and HNF3β signaling.
Adenomatous polyposis coli (APC)-regulated Wnt and transforming growth factor-β (TGFβ) signaling cooperate in the intestine to maintain normal enterocyte functions. Human clinical trials showed that estrogen [17β-estradiol (E2)], the ligand of nuclear receptors estrogen receptor α (ERα) and ERβ, inhibited colorectal cancer (CRC) in women. Consistent with this finding, we reported that E2, ERα and ERβ suppressed intestinal tumorigenesis in the C57BL/6J-Min/+ (Min/+) mouse, a CRC model. Here, we extended our results with further comparisons of colon and small intestine from intact female Apc (+/+) (WT), Min/+ and ER-deficient Min/+ mice. In the colon of ER-deficient Min/+ mice, ER loss reduced TGFβ signaling in crypt base cells as evidenced by minimal expression of the effectors Smad 2, 3 and 4 in these strains. We also found reduced expression of Indian hedgehog (Ihh), bone morphogenetic protein 4 and hepatocyte nuclear factor 3β or FoxA2, factors needed for paracrine signaling between enterocytes and mesenchyme. In proximal colon, ER loss produced a >10-fold increased incidence of crypt fission, a marker for wound healing and tumor promotion. These data, combined with our previous work detailing the specific roles of E2, ERα and ERβ in the colon, suggest that ER activity helps to maintain the intestinal stem cell (ISC) microenvironment by modulating epithelial-stromal crosstalk in ways that regulate cytokine, Wnt and Ihh availability in the extracellular matrix (ECM).


Induction of cyclooxygenase-2 by tumor promoters in transformed and cytochrome P450 2E1-expressing hepatocytes.
The induction of cyclooxygenase (COX)-2 expression has been implicated as a mechanism for the formation of non-hepatic tumors. Recent investigations have demonstrated COX-2 expression in human hepatocellular carcinomas, but little is known about the regulation of hepatocyte COX-2 expression. Employing the adult, rat hepatocyte line RALA255-10G, the effects of cellular transformation or expression of the alcohol-inducible cytochrome P450 2E1 (CYP2E1) on COX-2 expression were examined. Transformed and non-transformed hepatocytes did not express COX-2 by western and northern blot analysis. The tumor promoters phorbol 12-myristate 13-acetate (PMA) and chenodeoxycholic acid (CD) induced COX-2 protein expression in transformed, but not non-transformed cells. CYP2E1-expressing cells lacked constitutive COX-2 expression, and PMA but not CD induced COX-2 in these cells. PMA-treated transformed and CYP2E1-expressing cells expressed functional COX-2 as demonstrated by marked inductions in prostaglandin E(2) synthesis. PMA-induced COX-2 expression in both transformed and CYP2E1-expressing cells resulted from an induction in COX-2 mRNA, and was dependent on extracellular signal-regulated kinase, p38 mitogen-activated protein kinase and phosphatidylinositol 3-kinase. The differential induction of COX-2 by PMA in transformed and non-transformed cells could not be explained by differences in NF-kappaB or C/EBPalpha activation. PMA did not induce COX-2 transcriptional activity as determined by transient transfections with a luciferase reporter gene driven by the COX-2 promoter. The data demonstrate that cellular transformation and CYP2E1 expression fail to lead to the induction of COX-2 expression in hepatocytes. However, these conditions do render hepatocytes susceptible to COX-2 induction from tumor promoters by post-transcriptional mechanisms.


Increased Maternal Prenatal Adiposity, Inflammation, and Lower Omega-3 Fatty Acid Levels Influence Child Negative Affect.
Increased maternal adiposity during pregnancy is associated with offspring risk for psychiatric disorders. Inflammation secondary to adiposity is believed to be an important mechanism through which this effect occurs. Although increased adiposity introduces risk, not all children of overweight mothers develop these problems. Gestational factors that modify this risk are not well-understood. If maternal increased adiposity exerts its effects on offspring outcomes by increasing inflammation in the gestational environment, then anti-inflammatory inputs such as omega-3 fatty acids may be one protective factor. The goal of this study was to investigate whether maternal pre-pregnancy body mass index (BMI) and omega-3 fatty acid levels independently and/or interactively predicted offspring infant negative affect, an early life marker of risk for psychopathology. Data came from a prospective study of women recruited during pregnancy and their 6 month old infants (N = 62; 40% female). Maternal pre-pregnancy BMI was pulled from medical charts and third trimester omega-3 fatty acid concentrations were assessed in plasma. Child negative affect was assessed using observer- and maternal-ratings at 6 months of age. Maternal inflammation was indexed by third trimester plasma levels of interleukin-6, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1. Maternal pre-pregnancy BMI was associated with increased infant negative affect whereas eicosapentaenoic acid was associated with less infant negative affect. Maternal omega-3 fatty acid levels moderated the effect of BMI on infant negative affect, such that omega-3 fatty acids buffered children against the negative consequences of increased adiposity. Supporting the role of maternal inflammation in these associations, maternal BMI and omega-3 fatty acid levels interacted to predict maternal third trimester inflammation. Further, maternal inflammation was associated with increased infant negative affect. Results suggest that omega-3 supplementation during pregnancy may protect against offspring behavioral risk associated with increased maternal adiposity.


Mapping online hate: A scientometric analysis on research trends and hotspots in research on online hate.
Internet and social media participation open doors to a plethora of positive opportunities for the general public. However, in addition to these positive aspects, digital technology also provides an effective medium for spreading hateful content in the form of cyberbullying, bigotry, hateful ideologies, and harassment of individuals and groups. This research aims to investigate the growing body of online hate research (OHR) by mapping general research indices, prevalent themes of research, research hotspots, and influential stakeholders such as organizations and contributing regions. For this, we use scientometric techniques and collect research papers from the Web of Science core database published through March 2019. We apply a predefined search strategy to retrieve peer-reviewed OHR and analyze the data using CiteSpace software by identifying influential papers, themes of research, and collaborating institutions. Our results show that higher-income countries contribute most to OHR, with Western countries accounting for most of the publications, funded by North American and European funding agencies. We also observed increased research activity post-2005, starting from more than 50 publications to more than 550 in 2018. This applies to a number of publications as well as citations. The hotbeds of OHR focus on cyberbullying, social media platforms, co-morbid mental disorders, and profiling of aggressors and victims. Moreover, we identified four main clusters of OHR: (1) Cyberbullying, (2) Sexual solicitation and intimate partner violence, (3) Deep learning and automation, and (4) Extremist and online hate groups, which highlight the cross-disciplinary and multifaceted nature of OHR as a field of research. The research has implications for researchers and policymakers engaged in OHR and its associated problems for individuals and society.


Different doses of steroid injection in elderly patients with carpal tunnel syndrome: a triple-blind, randomized, controlled trial.
Carpal tunnel syndrome (CTS) is commonly seen in elderly populations, in part due to increased presence of predisposing comorbidities as well as physiological changes. We aimed at comparing the effectiveness of different doses of steroid using the ultrasound-guided hydrodissection method in elderly patients with CTS. We conducted a prospective, triple-blind, randomized, controlled trial in elderly patients with CTS. Patients were allocated to one of three groups by simplified randomization. Groups I-III received 80 mg triamcinolone (2 mL) and 1 mL of 2% lidocaine; 40 mg triamcinolone (1 mL), 1 mL of 2% lidocaine, and 1 mL normal saline; and 1 mL of 2% lidocaine and 2 mL normal saline, respectively to make up to 3 mL volume. A wrist splint was then applied for support. Outcome measures included the visual analog scale (VAS) and the Boston Carpal Tunnel Questionnaire, and median motor and sensory nerve conduction and its sonographic inlet cross-sectional area were used as objective measures. All data were recorded at baseline and 2, 12, and 24 weeks after injection. The investigators, patients, and statistician were blinded to the treatment assignment. In total, 161 patients were recruited without statistically significant demographic differences between the three groups. There were no statistically significant differences between groups in any outcome, with the exception of the median distal motor latency, which was greater in Group II at all three follow-up visits, and significant baseline VAS difference between Groups I and III. Hydrodissection with lidocaine and normal saline is as effective as hydrodissection with low- and high-dose steroid medication in elderly patients with CTS in this study, but further studies with matched baseline measures and also a sham group are suggested for definitive recommendation.


Outer membrane permeability in Pseudomonas aeruginosa: comparison of a wild-type with an antibiotic-supersusceptible mutant.
The Pseudomonas aeruginosa mutant Z61 has been shown to be highly supersusceptible to a wide range of antibiotics, including beta-lactams, aminoglycosides, rifampin, tetracycline, and chloramphenicol (W. Zimmerman, Int. J. Clin. Pharmacol. Biopharm. 17:131-134, 1979). Spontaneous revertants were isolated, using gentamicin or carbenicillin as selective agents, and shown to have two patterns of susceptibility to a group of 12 antibiotics. Partial revertants had 2- to 10-fold greater resistance to these antibiotics than mutant Z61, whereas full revertants had antibiotic susceptibilities indistinguishable from those of the wild-type strain K799, from which mutant Z61 had been derived. Uptake of a chromogenic beta-lactam nitrocefin was studied in both uninduced and induced cells of all strains by measuring the steady-state rate of nitrocefin hydrolysis by the inducible, periplasmic beta-lactamase in both whole and broken cells. This demonstrated that outer membrane permeability decreased as antibiotic resistance increased in the series mutant Z61, partial revertants, wild type, and full revertants. The data were consistent with the idea of low outer membrane permeability being caused by a low proportion of open functional porins in the outer membrane as the reason for the high natural antibiotic resistance of wild-type P, aeruginosa strains. In addition, it was observed that levels of benzylpenicillin below the minimal inhibitory concentration for mutant Z61 failed to induce beta-lactamase production. The possibility that this was related to the observed increase in outer membrane permeability is discussed. Preliminary evidence is presented that the pore-forming outer membrane porin protein F is not altered in mutant Z61.


A comparison of 2-octyl cyanoacrylate adhesives versus conventional suture materials for eyelid wound closure in rabbits.
To evaluate the clinical efficacy and histopathological tolerance of 2-octyl cyanoacrylate versus conventional suture materials for eyelid wound closure in rabbits. We performed an experimental study on 16 eyes of eight New Zealand albino rabbits. Eyelid incisions of 15 mm were done 4mm from the upper eyelid margin in both eyes. The eyes of the rabbits were divided into two groups: eyelid incisions of the right eye were closed by a 2-octyl cyanoacrylate adhesive (group A) and eyelid incisions of the left eye were closed by 7-0 nylon sutures (group B). At 1, 2, 4, and 8 weeks after surgery, the rabbits were macroscopically examined and then sacrificed. The specimens of their eyelid tissues were stained by a hematoxylin and eosin stain and Masson-trichrome stain, and were observed under microscope. Both eyelid surgical closure methods were found to be equally efficacious in fixing the eyelids of groups A and B, and their clinical efficacy was similar. Histopathological findings of the hematoxylin and eosin stain of group A showed less inflammatory infiltration than group B at 2 weeks. There were no significant histopathological differences between the two groups at 1, 4, and 8 weeks. The degree of fibrosis of the Masson-trichrome stain was similar between the two groups at 8 weeks. The 2-octyl cyanoacrylate adhesive proved to be an effective eyelid closure method and was very well tolerated by the skin surface. 2-Octyl cyanoacrylate could be used as an alternative tissue adhesive for eyelid wound closure along with conventional suture materials.


Localization of glycosaminoglycans in periodontal ligament during physiological and experimental tooth movement.
Localization of chondroitin sulphates in periodontal ligaments (PDL) of rat molar roots during physiological and experimental tooth movement were analysed immunohistochemically with the use of monoclonal antibodies, 3B3 and 2B6, specific to chondroitin 6-sulphate (CH-6S) and chondroitin 4-sulphate/dermatan sulfate (CH-4S/DS), respectively. The maxillary first molars of experimental animals were forced to move laterally with a 10 g weight by U-shaped wires for 3 and 7 d. In control animals, 3B3 epitope was seen in the PDL near to the bone surface facing the distal half of roots, which corresponded to the compressive side during physiological tooth movement. Immunoreactivity for 2B6 was weak or negative in the PDL. Both epitopes were present at osteoid, precementum, lacunae and canaliculli of osteocytes and cementocytes. In 3-d-treated animals, the early stage of hyalinization characterized with visible cells and fibres was observed in the PDL at the buccal side of the mesial root, which showed intense immunoreactivity for 3B3. Further 3B3 positive area seen in control animals changed its position from the distal to the buccal side of the PDL. Immunoreactivity for 2B6 did not change in the PDL of 3-d-treated animals. In 7-d-treated animals, the typical hyalinization characterized with no visible cells and fibres was seen in the PDL at the buccal sides of both mesial and disto-buccal roots, where both epitopes were present at the peripheral part of the tissue. Observation of serial sections suggested that the 3B3-positive area was present at the peripheral part of the 2B6-positive area. The present results suggest that the expression of CH-6S is related to the compressive force in non-hyalinized and hyalinized PDL, whereas that of CH-4S/DS is not influenced by the mechanical stress.


First Measurement of Energy-Dependent Inclusive Muon Neutrino Charged-Current Cross Sections on Argon with the MicroBooNE Detector.
We report a measurement of the energy-dependent total charged-current cross section σ(E_{ν}) for inclusive muon neutrinos scattering on argon, as well as measurements of flux-averaged differential cross sections as a function of muon energy and hadronic energy transfer (ν). Data corresponding to 5.3×10^{19} protons on target of exposure were collected using the MicroBooNE liquid argon time projection chamber located in the Fermilab booster neutrino beam with a mean neutrino energy of approximately 0.8 GeV. The mapping between the true neutrino energy E_{ν} and reconstructed neutrino energy E_{ν}^{rec} and between the energy transfer ν and reconstructed hadronic energy E_{had}^{rec} are validated by comparing the data and Monte Carlo (MC) predictions. In particular, the modeling of the missing hadronic energy and its associated uncertainties are verified by a new method that compares the E_{had}^{rec} distributions between data and a MC prediction after constraining the reconstructed muon kinematic distributions, energy, and polar angle to those of data. The success of this validation gives confidence that the missing energy in the MicroBooNE detector is well modeled and underpins first-time measurements of both the total cross section σ(E_{ν}) and the differential cross section dσ/dν on argon.


Diminished luteinizing hormone pulse frequency and amplitude with aging in the male rat.
Aging in the male rat is associated with a reduction in circulating testosterone levels. One possible cause of this decline is an age-related alteration of central nervous system-mediated LH secretion. To examine the effects of age on the hypothalamo-hypophyseal system, in the absence of gonadal steroid feedback, we studied the pattern of pulsatile LH secretion in castrate male Sprague-Dawley rats, aged 3 months (young), 8 months (middle-aged), and 26 months (old). All animals were castrated, and after 3 weeks, they were implanted with indwelling atrial catheters. One day later, duplicate 25 microliters blood samples were obtained at 4-min intervals for 4 h, while the animals were awake and unrestrained. Serum levels of LH, FSH, and testosterone were measured in animals before castration, and blood LH levels were measured in the postcastration, repeated sampling studies. After castration, middle-aged and old animals exhibited significantly lower mean serum LH levels, associated with a diminished amplitude of LH secretory episodes compared to young rats. In the oldest group, LH pulse frequency was significantly lower compared to middle-aged and young animals. Since the control of LH secretory episodes resides in the central nervous system, we propose that alterations in frequency of LH pulses observed in the aged, castrate male rat are the result of a diminished functional capacity of LHRH-containing neurons or of neurotransmitters that modulate their activity in the aging brain.


Immunotherapy of Childhood Sarcomas.
Pediatric sarcomas are a heterogeneous group of malignant tumors of bone and soft tissue origin. Although more than 100 different histologic subtypes have been described, the majority of pediatric cases belong to the Ewing's family of tumors, rhabdomyosarcoma and osteosarcoma. Most patients that present with localized stage are curable with surgery and/or chemotherapy; however, those with metastatic disease at diagnosis or those who experience a relapse continue to have a very poor prognosis. New therapies for these patients are urgently needed. Immunotherapy is an established treatment modality for both liquid and solid tumors, and in pediatrics, most notably for neuroblastoma and osteosarcoma. In the past, immunomodulatory agents such as interferon, interleukin-2, and liposomal-muramyl tripeptide phosphatidyl-ethanolamine have been tried, with some activity seen in subsets of patients; additionally, various cancer vaccines have been studied with possible benefit. Monoclonal antibody therapies against tumor antigens such as disialoganglioside GD2 or immune checkpoint targets such as CTLA-4 and PD-1 are being actively explored in pediatric sarcomas. Building on the success of adoptive T cell therapy for EBV-related lymphoma, strategies to redirect T cells using chimeric antigen receptors and bispecific antibodies are rapidly evolving with potential for the treatment of sarcomas. This review will focus on recent preclinical and clinical developments in targeted agents for pediatric sarcomas with emphasis on the immunobiology of immune checkpoints, immunoediting, tumor microenvironment, antibody engineering, cell engineering, and tumor vaccines. The future integration of antibody-based and cell-based therapies into an overall treatment strategy of sarcoma will be discussed.


Development of a syndromic surveillance system for detection of disease among livestock entering an auction market.
To develop a syndromic surveillance system based on visual inspection from outside the livestock pens that could be used for detection of disease among livestock entering an auction market. Cross-sectional study. All livestock (beef and dairy cattle, sheep, goats, horses, and pigs) entering a single auction market in Colorado during 30 business days. Procedures-Livestock were enumerated and visually inspected for clinical signs of disease by a veterinarian outside the pens, and clinical signs that were observed were categorized into 12 disease syndromes. Frequency of clinical signs and disease syndromes was then calculated. Data were recorded for a total of 29,371 animal observation days. For all species combined, the most common disease syndrome was respiratory tract disease (218.9 observations/10,000 animal observation days), followed by thin body condition and abnormal ambulation or posture (80.7 and 27.2 observations/10,000 animal observation days, respectively). Together, these 3 disease syndromes accounted for 92.8% of all clinical signs of disease observed. The syndromes least commonly identified were non-injury-related hemorrhage, death, and injury-related hemorrhage (0.0, 0.3, and 0.7 observations/10,000 animal observation days, respectively). Results suggested that a syndromic surveillance system based on visual inspection alone could be developed to identify possible disease conditions among livestock at an auction market. Further studies are needed to determine the sensitivity and specificity of visual observation in detecting disease.


Inorganic carbon assimilation in the Isoetids, Isoetes lacustris L. and Lobelia dortmanna L.
The inorganic carbon fixation patterns of Isoetes lacustris and Lobelia dortmanna from an oligotrophic Scottish loch have been examined by following titratable acidity changes in plant sap and light/dark 14CO2 incorporation by roots and shoots. The diurnal pattern of titratable acidity changes in I. lacustris suggests crassulacean acid metabolism (CAM) while the lack of any change in titratable acidity in L. dortmanna suggests C3 metabolism. Of the carbon fixed by L. dortmanna, 99.9% was taken up through the roots and fixation occurred primarily during the day. In Isoetes, CO2 was taken up by both roots and shoots and during both day and night. Regardless of the site of CO2 uptake, fixation occurred only in the shoots of both plants. Analysis of carbon isotope ratios of plant organic material was used to further investigate the photosynthetic mechanisms of these Isoetids. Considering the absence of a nighttime peak in titratable acidity in L. dortmanna, the Δ13C (Δ=δ13C plant-δ13C source) value of the shoots of L. dortmanna (-14.2‰) is indicative of C3 photosynthesis limited by the rate of CO2 diffusion. The less negative Δ of I. lacustris (-6.0‰) is consistent with both dark acidification of CAM and CO2 limited C3 photosynthesis. This is in contrast to the terrestrial Isoetes durieui which is shown to have a Δ value which is similar to a terrestrial C3 plant. The carbon fixation patterns of these Isoetids suggest that the CO2 concentration in the loch may be growth limiting, and that root uptake and/or dark acidification are means of optimising CO2 supply. However, in view of the relatively high levels of CO2 in sediment and bulk water, it is suggested that low levels of nutrients may also limit growth in these plants.


The Prevalence of Nonprescription Cannabinoid-Based Medicines in British Children's Hospices: Results of a National Survey.
Background: Almost 50,000 children and young people are affected by life-limiting conditions in the United Kingdom, around a third of which use children's hospices. Anecdotal evidence suggests that cannabinoid-based medicines (CBMs), specifically cannabis oil (CO), are being used by families with increasing frequency to manage distressing symptoms. The use of most nonprescription CBMs in the United Kingdom remains illegal. Objective: The objective of the study was to identify the prevalence of CO use by families who use children's hospices in the United Kingdom, and the approaches taken by those services to manage it. Design: An electronic survey was sent to each of the 54 children's hospices in the United Kingdom between May and July 2018, comprising 10 questions. Results: Forty children's hospices from across the four countries of the United Kingdom responded to the survey, representing 74% of British children's hospices. About 87.5% of hospices knew of children who use CO therapeutically. Sixty-nine percent of those hospices have received requests to administer CO during an episode of care. Approaches by organizations around CO management varied across the sectors, including arrangements for storage, administration, and recording of its use. Hospices highlighted how the lack of available guidance made decision making more challenging. Only a third of responding organizations routinely questioned families about the use of cannabis when prescribing medicines. Conclusion: CO is used extensively by children who use children's hospices. Despite recognizing the use of CO, many hospices are unable to support it. There is a need for clear guidelines on how hospices should approach the care needs of children, allowing hospices to meet the needs of children who use CO, and families in a safe, consistent, and relevant way, safeguarding all children, families, and professionals within the organization.


Anthropometric standardisation and quality control protocols for the construction of new, international, fetal and newborn growth standards: the INTERGROWTH-21st Project.
The primary aim of the INTERGROWTH-21(st) Project is to construct new, prescriptive standards describing optimal fetal and preterm postnatal growth. The anthropometric measurements include the head circumference, recumbent length and weight of the infants, and the stature and weight of the parents. In such a large, international, multicentre project, it is critical that all study sites follow standardised protocols to ensure maximal validity of the growth and nutrition indicators used. This paper describes in detail the anthropometric training, standardisation and quality control procedures used to collect data for these new standards. The initial standardisation session was in Nairobi, Kenya, using newborns, which was followed by similar sessions in the eight participating study sites in Brazil, China, India, Italy, Kenya, Oman, UK and USA. The intraobserver and inter-observer technical error of measurement values for head circumference range from 0.3 to 0.4 cm, and for recumbent length from 0.3 to 0.5 cm. These standardisation protocols implemented at each study site worldwide ensure that the anthropometric data collected are of the highest quality to construct international growth standards.


Genetic evaluation of days to harvest in crossbred lambs.
Days to harvest (DTH) is the number of days a lamb is fed before reaching a target level of fatness. Although economically relevant, this trait has not been thoroughly evaluated in sheep. Most lambs harvested in the United Kingdom are crossbreds sired by purebred terminal sires, with Charollais, Suffolk, and Texel most commonly used. Sires from these breeds were selected on an index designed to increase lean growth while constraining fat. The purpose of this research was to 1) evaluate the effects of index selection in terminal sires on DTH and 2) evaluate the feasibility of incorporating DTH into genetic evaluation programs. Charollais, Suffolk, and Texel sheep had participated in sire referencing schemes where genetic links among flocks were established by sharing rams. Rams with high or low index scores were chosen from these schemes and mated to crossbred ewes at 3 farms in the United Kingdom. Lambs were harvested at a target 11% subcutaneous fat. Records on DTH from 6,350 lambs were analyzed in 2 ways: 1) as time to harvest fitting a survival model and 2) as a normally distributed variable in a bivariate analysis with weight at harvest. The survival analysis was stratified by rearing type (single or twin). In both approaches, sires were fitted using a multivariate normal distribution with a relationship matrix. Regardless of model fitted, sire index did not affect DTH (P>0.10). However, Texel-sired lambs reached harvest faster (P<0.01) than either Charollais- or Suffolk-sired lambs although DTH in those 2 breed types did not differ (P>0.1). Ewe lambs reached harvest faster than wethers (P<0.01). Lambs from older ewes were harvested faster (P<0.001). The heritability of DTH was 0.21 from the survival model and 0.20 from the bivariate model. Rank correlation of sire EBV between methods was 0.9, suggesting strong agreement. The use of high or low index sires did not extend DTH in lambs harvested at a target fatness. Importantly, there is no antagonism between improving carcass merit and extending the grazing season. Furthermore, DTH is moderately heritable. If economically justified within a breeding program, it could be reduced through genetic selection.


Antagonists of protein kinase C inhibit rat retinal glutamate transport activity in situ.
Neuronal and glial high-affinity transporters regulate extracellular glutamate concentration, thereby terminating synaptic transmission and preventing neuronal excitotoxicity. Glutamate transporter activity has been shown to be modulated by protein kinase C (PKC) in cell culture. This is the first study to demonstrate such modulation in situ, by following the fate of the non-metabolisable glutamate transporter substrate, d-aspartate. In the rat retina, pan-isoform PKC inhibition with chelerythrine suppressed glutamate uptake by GLAST (glutamate/aspartate transporter), the dominant excitatory amino acid transporter localized to the glial Müller cells. This effect was mimicked by rottlerin but not by Gö6976, suggesting the involvement of the PKCdelta isoform, but not PKCalpha, beta or gamma. Western blotting and immunohistochemical labeling revealed that the suppression of glutamate transport was not due to a change in transporter expression. Inhibition of PKCdelta selectively suppressed GLAST but not neuronal glutamate transporter activity. These data suggest that the targeting of specific glutamate transporters with isoform-specific modulators of PKC activity may have significant implications for the understanding of neurodegenerative conditions arising from compromised glutamate homeostasis, e.g. glaucoma and amyotrophic lateral sclerosis.


Seat back and head restraint response during low-speed rear-end automobile collisions.
Automobile seat backs and head restraints play a key safety role during low-speed rear-end collisions, yet few studies have explored the effect of collision variables on seat response. In this study, the effects of vehicle speed change and seat belt use on dynamic seat back and head restraint response during low-speed rear-end automobile collisions were examined. Four human subjects were repeatedly exposed to vehicle-to-vehicle rear-end collisions with speed changes of 2, 4, 6 and 8 km/h. Seat back force and deflection, and head restraint force were measured. The point of application of the resultant force applied to the seat back and head restraint were determined. The magnitude and time of peak kinematic and kinetic response parameters were used in a two-way repeated-measures analysis of variance (ANOVA) for speed change and seat belt use. The results showed that 20 of the 24 seat back and head restraint response parameters varied with speed change and none of the parameters varied with seat belt use. Head restraint forces, seat back forces and seat back deflections increased approximately linearly with speed change, whereas time to peak response, direction and moment arm of the forces remained either constant or varied only slightly over the range of speed changes tested.


Characteristics and prediction of RNA editing sites in transcripts of the Moss Takakia lepidozioides chloroplast.
RNA editing in land plant organelles is a process primarily involving the conversion of cytidine to uridine in pre-mRNAs. The process is required for gene expression in plant organelles, because this conversion alters the encoded amino acid residues and improves the sequence identity to homologous proteins. A recent study uncovered that proteins encoded in the nuclear genome are essential for editing site recognition in chloroplasts; the mechanisms by which this recognition occurs remain unclear. To understand these mechanisms, we determined the genomic and cDNA sequences of moss Takakia lepidozioides chloroplast genes, then computationally analyzed the sequences within -30 to +10 nucleotides of RNA editing sites (neighbor sequences) likely to be recognized by trans-factors. As the T. lepidozioides chloroplast has many RNA editing sites, the analysis of these sequences provides a unique opportunity to perform statistical analyses of chloroplast RNA editing sites. We divided the 302 obtained neighbor sequences into eight groups based on sequence similarity to identify group-specific patterns. The patterns were then applied to predict novel RNA editing sites in T. lepidozioides transcripts; approximately 60% of these predicted sites are true editing sites. The success of this prediction algorithm suggests that the obtained patterns are indicative of key sites recognized by trans-factors around editing sites of T. lepidozioides chloroplast genes.


Feasibility and effectiveness of the baby friendly community initiative in rural Kenya: study protocol for a randomized controlled trial.
Interventions promoting optimal infant and young child nutrition could prevent a fifth of under-5 deaths in countries with high mortality. Poor infant and young child feeding practices are widely documented in Kenya, with potential detrimental effects on child growth, health and survival. Effective strategies to improve these practices are needed. This study aims to pilot implementation of the Baby Friendly Community Initiative (BFCI), a global initiative aimed at promoting optimal infant and young child feeding practices, to determine its feasibility and effectiveness with regards to infant feeding practices, nutrition and health outcomes in a rural setting in Kenya. The study, employing a cluster-randomized trial design, will be conducted in rural Kenya. A total of 12 clusters, constituting community units within the government's Community Health Strategy, will be randomized, with half allocated to the intervention and the other half to the control arm. A total of 812 pregnant women and their respective children will be recruited into the study. The mother-child pairs will be followed up until the child is 6 months old. Recruitment will last approximately 1 year from January 2015, and the study will run for 3 years, from 2014 to 2016. The intervention will involve regular counseling and support of mothers by trained community health workers and health professionals on maternal, infant and young child nutrition. Regular assessment of knowledge, attitudes and practices on maternal, infant and young child nutrition will be done, coupled with assessment of nutritional status of the mother-child pairs and morbidity for the children. Statistical methods will include analysis of covariance, multinomial logistic regression and multilevel modeling. The study is funded by the NIH and USAID through the Program for Enhanced Research (PEER) Health. Findings from the study outlined in this protocol will inform potential feasibility and effectiveness of a community-based intervention aimed at promoting optimal breastfeeding and other infant feeding practices. The intervention, if proved feasible and effective, will inform policy and practice in Kenya and similar settings, particularly regarding implementation of the baby friendly community initiative. ISRCTN03467700 ; Date of Registration: 24 September 2014.


Impact of 6-month frozen storage of cervical specimens in alkaline buffer conditions on human papillomavirus genotyping.
The impact of 6-month storage of cervical specimens under alkaline conditions that occurs as the result of Hybrid Capture 2 testing on human papillomavirus (HPV) genotyping is not well documented. To examine this issue, 143 frozen hc2-positive specimens in specimen transport medium were selected at random from each of the following groups: specimens stored for 6 months, 4 months, and 2.5 months under alkaline pH (pH 12-13) and specimens stored 1 month at neutral pH (pH 6-7) as controls. Specimens were tested in a masked fashion for 20 HPV genotypes (HPV6, 11, 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, and 82) using a prototype, research-use-only GP5+/6+ L1 consensus PCR method and multiplex hybridization using Luminex xMAP for detection of specific HPV genotypes One control specimen had missing test results. There were no statistical differences in the number of HPV genotypes detected, number of carcinogenic HPV genotypes detected, or in the signal strength among HPV-positive results across groups. Six-month frozen storage of cervical specimens at alkaline pH had little impact on testing for HPV genotypes among hc2-positive women using this HPV genotyping method.


Obstetric risk avoidance: will anyone be offering obstetrics in private practice by the end of the decade?
Obstetric claims inflation is increasing the cost of covering obstetric risk. This is leading to obstetric risk avoidance by those offering insurance and by practitioners who do not perform enough deliveries to cover the cost of obstetric risk indemnity. By the end of the decade indemnifying obstetric risk will probably be too expensive for doctors in private practice. Non-indemnified doctors will be unable or unwilling to do private deliveries; however, women will still fall pregnant and require delivery. These women will inevitably be forced to deliver in provincial facilities, shifting the workload and liability to the state.


Na+/H+ exchanger regulatory factor-1 is involved in chemokine receptor homodimer CCR5 internalization and signal transduction but does not affect CXCR4 homodimer or CXCR4-CCR5 heterodimer.
Chemokine receptors are members of the G protein-coupled receptor (GPCR) family. CCR5 is also the principal co-receptor for macrophage-tropic strains of human immunodeficiency virus, type 1 (HIV-1), and efforts have been made to develop ligands to inhibit HIV-1 infection by promoting CCR5 receptor endocytosis. Given the nature of GPCRs and their propensity to form oligomers, one can consider ligand-based therapies as unselective in terms of the oligomeric composition of complexes. For example, a ligand targeting a CCR5 homomer could likely induce signal transduction on a heteromeric CCR5-CXCR4. Other avenues could therefore be explored. We identified a receptor adaptor interacting specifically with one receptor complex but not others. NHERF1, an adaptor known for its role in desensitization, internalization, and regulation of the ERK signaling cascade for several GPCRs, interacts via its PDZ2 domain with the CCR5 homodimer but not with the CXCR4-CCR5 heterodimer or CXCR4 homodimer. To further characterize this interaction, we also show that NHERF1 increases the CCR5 recruitment of arrestin2 following stimulation. NHERF1 is also involved in CCR5 internalization, as we demonstrate that co-expression of constructs bearing the PDZ2 domain can block CCR5 internalization. We also show that NHERF1 potentiates RANTES (regulated on activation normal T cell expressed and secreted)-induced ERK1/2 phosphorylation via CCR5 activation and that this activation requires NHERF1 but not arrestin2. Taken together, our results suggest that oligomeric receptor complexes can associate specifically with partners and that in this case NHERF1 could represent an interesting new target for the regulation of CCR5 internalization and potentially HIV infection.


Chromosome instability in lymphocytes: a potential indicator of predisposition to oral premalignant lesions.
Oral premalignant lesions (OPLs) are related to tobacco use and mark individuals at high risk for oral cancer development. Increased mutagen sensitivity as measured by an in vitro mutagen challenge assay has been shown to be a risk factor for upper aerodigestive tract cancers. In this case control study, we used two assays with mutagens relevant to tobacco exposure (benzo[a]pyrene diol epoxide (BPDE) and bleomycin) to see whether sensitivity to these mutagens could be used as biomarkers for assessing risk of premalignant lesions. Furthermore, we evaluated whether 3p21.3 is a molecular target of BPDE damage in lymphocytes of patients with OPLs. There were 82 patients with OPLs and 89 healthy controls frequency matched to the cases on age, sex, ethnicity, and smoking status. These subjects' lymphocytes were treated in two separate experiments with either 2 microM BPDE for 24 h or 0.03 units/ml bleomycin for 5 h, and the frequency of induced chromatid breakage in Giemsa-stained preparations was determined. BPDE-induced 3p21.3 aberrations were scored by fluorescent in situ hybridization technique in 1000 interphases/sample. We found that the mean BPDE-induced chromatid breaks per cell were higher in cases than controls (1.05 +/- 0.40 and 0.55 +/- 0.27, respectively; P < 0.01). Similar results were evident with bleomycin-induced chromatid breaks per cell (0.78 +/- 0.37 and 0.57 +/- 0.31, respectively; P < 0.01). After adjusting for age, sex, ethnicity, and smoking status, significantly elevated odds ratios (95% confidence interval) for OPL risk were noted for BPDE sensitivity [12.96 (5.51, 30.46)] and bleomycin sensitivity [3.33 (1.64, 6.77)]. When subjects were categorized into quartiles of the number of breaks per cell, a dose response was observed for both assays. The adjusted odds ratios for subjects with increasing numbers of breaks per cell in quartiles were 2.34, 9.14, and 54.04 for BPDE sensitivity and 1.92, 3.33, and 7.15 for bleomycin sensitivity, respectively. Subjects sensitive to both mutagens had a 50-fold increased risk for OPLs. In addition, there were significantly more BPDE-induced chromosome aberrations at the 3p21.3 locus in cases (51.13/1000) than in controls (40.93/1000; P < 0.0001). However, no such difference was observed for 3q13, a control locus. BPDE-induced 3p21.3 aberrations were associated with an elevated risk for OPLs of 6.08 (2.57, 14.4). The degree of BPDE sensitivity at 3p21.3 and risk for OPLs increased in a dose-dependent manner. In summary, BDPE sensitivity and bleomycin sensitivity appear to be individually and jointly associated with elevated risk of OPLs. Furthermore, 3p21.3 may be a molecular target of BPDE in OPLs. This is the first study to examine mutagen sensitivity in a premalignant condition. The next step is to correlate these findings in surrogate (lymphocyte) tissue with molecular events in the target tissue.


Stroke patients' awareness of risk and readiness to change behaviors.
Behavior change is an important component of secondary stroke prevention. The transtheoretical model, which describes behavior change as occurring through a series of stages, may be a useful way of assessing patients' readiness to change behavior. The model has been successfully applied to other chronic conditions and argues that people progressing "forward" through the stages are more likely to successfully change their behavior. The aim of this study was to describe stroke patients' readiness to change behaviors for stroke-related risk factors using this model, in the absence of a behavior modification intervention. Patients (n = 27) from an acute stroke ward of a major metropolitan hospital in Brisbane, Australia, were interviewed prior to and at 3 months following hospital discharge regarding their awareness of stroke risk factors and their readiness to change stroke risk-related behaviors. At both points in time, 30% of patients could not spontaneously nominate one or more stroke risk factors. Despite a trend of "forward" progression in stages of change between the 2 interviews for behaviors relating to hypertension, heart disease, and high cholesterol, there were no statistically significant changes over time for any of the behaviors. Patients' readiness to change stroke risk-related behaviors differed for each risk factor. Acknowledging that patients' readiness to change may differ for each risk factor may promote more effective facilitation of stroke secondary prevention behaviors.


Accuracy of genotype imputation in Labrador Retrievers.
The dog is a valuable model species for the genetic analysis of complex traits, and the use of genotype imputation in dogs will be an important tool for future studies. It is of particular interest to analyse the effect of factors like single nucleotide polymorphism (SNP) density of genotyping arrays and relatedness between dogs on imputation accuracy due to the acknowledged genetic and pedigree structure of dog breeds. In this study, we simulated different genotyping strategies based on data from 1179 Labrador Retriever dogs. The study involved 5826 SNPs on chromosome 1 representing the high density (HighD) array; the low-density (LowD) array was simulated by masking different proportions of SNPs on the HighD array. The correlations between true and imputed genotypes for a realistic masking level of 87.5% ranged from 0.92 to 0.97, depending on the scenario used. A correlation of 0.92 was found for a likely scenario (10% of dogs genotyped using HighD, 87.5% of HighD SNPs masked in the LowD array), which indicates that genotype imputation in Labrador Retrievers can be a valuable tool to reduce experimental costs while increasing sample size. Furthermore, we show that genotype imputation can be performed successfully even without pedigree information and with low relatedness between dogs in the reference and validation sets. Based on these results, the impact of genotype imputation was evaluated in a genome-wide association analysis and genomic prediction in Labrador Retrievers.


Systemic amyloidosis: lessons from β2-microglobulin.
β2-Microglobulin is responsible for systemic amyloidosis affecting patients undergoing long-term hemodialysis. Its genetic variant D76N causes a very rare form of familial systemic amyloidosis. These two types of amyloidoses differ significantly in terms of the tissue localization of deposits and for major pathological features. Considering how the amyloidogenesis of the β2-microglobulin mechanism has been scrutinized in depth for the last three decades, the comparative analysis of molecular and pathological properties of wild type β2-microglobulin and of the D76N variant offers a unique opportunity to critically reconsider the current understanding of the relation between the protein's structural properties and its pathologic behavior.


Molecular cloning of chromogranin A from rat pheochromocytoma cells.
Chromogranin A (CgA) is the major soluble protein in catecholamine storage vesicles. To gain insight into its function, we isolated CgA clones from a size-selected lambda gt10 rat pheochromocytoma complementary DNA (cDNA) library. The longest cDNA insert identified was 2.2 kb and encoded the entire 462-amino acid open reading frame of rat CgA including an 18-amino acid hydrophobic signal peptide. Comparison of rat CgA with the recently published sequences of bovine CgA and human CgA revealed regions of strong homology at the N-and COOH-termini as well as variant areas predominantly in the middle portion of the molecule. Regions highly conserved and therefore suggestive of functional importance included 1) multiple paired basic residues, which may serve as proteolytic processing signals; 2) a region homologous to porcine pancreastatin, a putative modulator of peptide hormone release; and 3) a short hydrophobic disulfide loop region near the N-terminus that may have a role in the targeting of CgA to secretory vesicles. On the other hand, lack of conservation of the membrane attachment sequence arginine-glycine-aspartic acid argues against its functional importance in CgA. In addition, the presence of a unique polyglutamine region in rat CgA points to a possible messenger RNA (mRNA) splice junction. Northern blot experiments demonstrated the presence of an approximately 2.2 kb rat CgA mRNA in a neuroendocrine distribution (adrenal, brain, pheochromocytoma cells, but not skeletal muscle, heart, or kidney). Southern blot studies were consistent with the presence of a single CgA gene within the rat pheochromocytoma cell genome. Finally, comparison of the present rat pheochromocytoma cDNA clones with those recently obtained from normal rat adrenal gland reveals minor but apparently real differences that suggest CgA microheterogeneity.


Inverse-Weighted Survival Games.
Deep models trained through maximum likelihood have achieved state-of-the-art results for survival analysis. Despite this training scheme, practitioners evaluate models under other criteria, such as binary classification losses at a chosen set of time horizons, e.g. Brier score (BS) and Bernoulli log likelihood (BLL). Models trained with maximum likelihood may have poor BS or BLL since maximum likelihood does not directly optimize these criteria. Directly optimizing criteria like BS requires inverse-weighting by the censoring distribution. However, estimating the censoring model under these metrics requires inverse-weighting by the failure distribution. The objective for each model requires the other, but neither are known. To resolve this dilemma, we introduce Inverse-Weighted Survival Games. In these games, objectives for each model are built from re-weighted estimates featuring the other model, where the latter is held fixed during training. When the loss is proper, we show that the games always have the true failure and censoring distributions as a stationary point. This means models in the game do not leave the correct distributions once reached. We construct one case where this stationary point is unique. We show that these games optimize BS on simulations and then apply these principles on real world cancer and critically-ill patient data.


Chimpanzee and felid diet composition is influenced by prey brain size.
Prey use a wide variety of anti-predator defence strategies, including morphological and chemical defences as well as behavioural traits (risk-modulated habitat use, changes in activity patterns, foraging decisions and group living). The critical test of how effective anti-predator strategies are is to relate them to relative indices of mortality across predators. Here, we compare biases in predator diet composition with prey characteristics and show that chimpanzee (Pan troglodytes) and felid show the strongest and the most consistent predator bias towards small-brained prey. We propose that large-brained prey are likely to be more effective at evading predators because they can effectively alter their behavioural responses to specific predator encounters. Thus, we provide evidence for the hypothesis that brain size evolution is potentially driven by selection for more sophisticated and behaviourally flexible anti-predator strategies.


Disruption of the FMN-A524 interaction cascade and Glu513-induced collapse of the hydrophobic barrier promotes light-induced Jα-helix unfolding in AsLOV2.
The C-terminal Jα-helix of the Avena sativa's Light Oxygen and Voltage (AsLOV2) protein, unfolds on exposure to blue light. This characteristic seeks relevance in applications related to engineering novel biological photoswitches. Using molecular dynamics simulations and the Markov state modeling (MSM) approach we provide the mechanism that explains the stepwise unfolding of the Jα-helix. The unfolding was resolved into seven steps represented by the structurally distinguishable states distributed over the initiation and the post initiation phases. Whereas, the initiation phase occurs due to the collapse of the interaction cascade FMN-Q513-N492-L480-W491-Q479-V520-A524, the onset of the post initiation phase is marked by breaking of the hydrophobic interactions between the Jα-helix and the Iβ-strand. This study indicates that the displacement of N492 out of the FMN binding pocket, not necessarily requiring Q513, is essential for the initiation of the Jα-helix unfolding. Rather, the structural reorientation of Q513 activates the protein to cross the hydrophobic barrier and enter the post initiation phase. Similarly, the structural deviations in N482, rather than its integral role in unfolding, could enhance the unfolding rates. Furthermore, the MSM studies on the wild-type and the Q513 mutant, provide the spatiotemporal roadmap that lay out the possible pathways of structural transition between the dark and the light states of the protein. Overall, the study provides insights useful to enhance the performance of AsLOV2-based photoswitches.