[Estimating probabilities and dealing with mutations in paternity testing--verification of DNA testing with commercially available STR kits].
Since Jeffreys devised a DNA fingerprint in 1985, DNA analysis has been applied to paternity testing. The progress of the techniques, especially the development of polymerase chain reaction (PCR), makes it possible to type 10-15 short tandem repeat (STR) loci in paternity testing by a single test tube. When using a well qualified database, we can now obtain a paternity index (PI) as high as 10(6) in usual trio cases. Furthermore, the DNA testings are now applied to unusual cases, such as personal identification of Japanese war orphans left in China. Here, we reviewed how to calculate the PI likelihood ratio and exclusion probability in a trio case, a motherless case, parent identification without reliable evidence of mother and child relationship, and a sibling case. We also reviewed how to handle single exclusion cases usually derived from a single mutation that is no longer rare when many STR loci are used. Finally, we emphasized the importance of ethical, legal and social counseling for clients in paternity testing. From that point of view, paternity tests by mail should not be allowed because of lack of such counseling.


Genetic diversity and family groups detected in a coyote population with red wolf ancestry on Galveston Island, Texas.
Hybridization can be a conservation concern if genomic introgression leads to the loss of an endangered species' unique genome, or when hybrid offspring are sterile or less fit than their parental species. Yet hybridization can also be an adaptive management tool if rare populations are inbred and have reduced genetic variation, and there is the opportunity to enhance genetic variation through hybridization. The red wolf (Canis rufus) is a critically endangered wolf endemic to the eastern United States, where all extant red wolves are descended from 14 founders which has led to elevated levels of inbreeding over time. Red wolves were considered extirpated from the wild by 1980, but before they disappeared, they interbred with encroaching coyotes creating a genetically admixed population of canids along coastal Texas and Louisiana. In 2018, a genetic study identified individuals on Galveston Island, Texas with significant amounts of red wolf ancestry. We collected 203 fecal samples from Galveston for a more in-depth analysis of this population to identify the amount of red wolf ancestry present and potential mechanisms that support retention of red wolf ancestry on the landscape. We identified 24 individual coyotes from Galveston Island and 8 from mainland Texas with greater than 10% red wolf ancestry. Two of those individuals from mainland Texas had greater than 50% red wolf ancestry estimates. Additionally, this population had 5 private alleles that were absent in the North American reference canid populations used in this study, which included 107 southeastern coyotes, 19 captive red wolves, and 38 gray wolves, possibly representing lost red wolf genetic variation. We also identified several individuals on Galveston Island and the mainland of Texas that retained a unique red wolf mitochondrial haplotype present in the red wolf founding population. On Galveston Island, we identified a minimum of four family groups and found coyotes on the island to be highly related, but not genetically depauperate. We did not find clear associations between red wolf ancestry estimates and landscape features, such as open green space or developed areas. Our results confirm the presence of substantial red wolf ancestry persisting on Galveston Island and adjacent mainland Texas. This population has the potential to benefit future red wolf conservation efforts through novel reproductive techniques and possibly through de-introgression strategies, with the goals of recovering extinct red wolf genetic variation and reducing inbreeding within the species.


Compromised fidelity of B-cell tolerance checkpoints in AChR and MuSK myasthenia gravis.
Myasthenia gravis (MG) is an autoimmune condition in which neurotransmission is impaired by binding of autoantibodies to acetylcholine receptors (AChR) or, in a minority of patients, to muscle specific kinase (MuSK). There are differences in the dominant IgG subclass, pathogenic mechanisms, and treatment responses between the two MG subtypes (AChR or MuSK). The antibodies are thought to be T-cell dependent, but the mechanisms underlying their production are not well understood. One aspect not previously described is whether defects in central and peripheral tolerance checkpoints, which allow autoreactive B cells to accumulate in the naive repertoire, are found in both or either form of MG. An established set of assays that measure the frequency of both polyreactive and autoreactive B cell receptors (BCR) in naive populations was applied to specimens collected from patients with either AChR or MuSK MG and healthy controls. Radioimmuno- and cell-based assays were used to measure BCR binding to AChR and MuSK. The frequency of polyreactive and autoreactive BCRs (n = 262) was higher in both AChR and MuSK MG patients than in healthy controls. None of the MG-derived BCRs bound AChR or MuSK. The results indicate that both these MG subtypes harbor defects in central and peripheral B cell tolerance checkpoints. Defective B cell tolerance may represent a fundamental contributor to autoimmunity in MG and is of particular importance when considering the durability of myasthenia gravis treatment strategies, particularly biologics that eliminate B cells.


The "island rule" and deep-sea gastropods: re-examining the evidence.
One of the most intriguing patterns in mammalian biogeography is the "island rule", which states that colonising species have a tendency to converge in body size, with larger species evolving decreased sizes and smaller species increased sizes. It has recently been suggested that an analogous pattern holds for the colonisation of the deep-sea benthos by marine Gastropoda. In particular, a pioneering study showed that gastropods from the Western Atlantic showed the same graded trend from dwarfism to gigantism that is evident in island endemic mammals. However, subsequent to the publication of the gastropod study, the standard tests of the island rule have been shown to yield false positives at a very high rate, leaving the result open to doubt. The evolution of gastropod body size in the deep sea is reexamined. Using an extended and updated data set, and improved statistical methods, it is shown that some results of the previous study may have been artifactual, but that its central conclusion is robust. It is further shown that the effect is not restricted to a single gastropod clade, that its strength increases markedly with depth, but that it applies even in the mesopelagic zone. The replication of the island rule in a distant taxonomic group and a partially analogous ecological situation could help to uncover the causes of the patterns observed--which are currently much disputed. The gastropod pattern is evident at intermediate depths, and so cannot be attributed to the unique features of abyssal ecology.


Thresholds of microwave-evoked warmth sensations in human skin.
We measured thresholds for microwave-evoked skin sensations of warmth at frequencies of 2.45, 7.5, 10, 35, and 94 GHz. In the same subjects, thresholds of warmth evoked by infrared radiation (IR) were also measured for comparison. Detection thresholds were measured on the skin in the middle of the back in 15 adult male human subjects at all microwave (MW) frequencies and with IR. Long duration (10-s), large area (327-cm2) stimuli were used to minimize any differential effects of temporal or spatial summation. Sensitivity increased monotonically with frequency throughout the range of microwave frequencies tested. The threshold at 94 GHz (4.5 +/- 0.6 mW/cm2) was more than an order of magnitude less than at 2.45 GHz (63.1 +/- 6.7 mW/cm2), and it was comparable to the threshold for IR (5.34 +/- 1.07 mW/cm2).


Type IV Pili in Francisella - A Virulence Trait in an Intracellular Pathogen.
Francisella tularensis is a highly virulent intracellular human pathogen that is capable of rapid proliferation in the infected host. Mutants affected in intracellular survival and growth are highly attenuated which highlights the importance of the intracellular phase of the infection. Genomic analysis has revealed that Francisella encodes all genes required for expression of functional type IV pili (Tfp), and in this focused review we summarize recent findings regarding this system in the pathogenesis of tularemia. Tfp are dynamic adhesive structures that have been identified as major virulence determinants in several human pathogens, but it is not obvious what role these structures could have in an intracellular pathogen like Francisella. In the human pathogenic strains, genes required for secretion and assembly of Tfp and one pilin, PilA, have shown to be required for full virulence. Importantly, specific genetic differences have been identified between the different Francisella subspecies where in the most pathogenic type A variants all genes are intact while several Tfp genes are pseudogenes in the less pathogenic type B strains. This suggests that there has been a selection for expression of Tfp with different properties in the different subspecies. There is also a possibility that the genetic differences reflect adaptation to different environmental niches of the subspecies and plays a role in transmission of tularemia. This is also in line with recent findings where Tfp pilins are found to be glycosylated which could reflect a role for Tfp in the environment to promote survival and transmission. We are still far from understanding the role of Tfp in virulence and transmission of tularemia, but with the genomic information and genetic tools available we are in a good position to address these issues in the future.


Capturing dopaminergic modulation and bimodal membrane behaviour of striatal medium spiny neurons in accurate, reduced models.
Loss of dopamine from the striatum can cause both profound motor deficits, as in Parkinson's disease, and disrupt learning. Yet the effect of dopamine on striatal neurons remains a complex and controversial topic, and is in need of a comprehensive framework. We extend a reduced model of the striatal medium spiny neuron (MSN) to account for dopaminergic modulation of its intrinsic ion channels and synaptic inputs. We tune our D1 and D2 receptor MSN models using data from a recent large-scale compartmental model. The new models capture the input-output relationships for both current injection and spiking input with remarkable accuracy, despite the order of magnitude decrease in system size. They also capture the paired pulse facilitation shown by MSNs. Our dopamine models predict that synaptic effects dominate intrinsic effects for all levels of D1 and D2 receptor activation. We analytically derive a full set of equilibrium points and their stability for the original and dopamine modulated forms of the MSN model. We find that the stability types are not changed by dopamine activation, and our models predict that the MSN is never bistable. Nonetheless, the MSN models can produce a spontaneously bimodal membrane potential similar to that recently observed in vitro following application of NMDA agonists. We demonstrate that this bimodality is created by modelling the agonist effects as slow, irregular and massive jumps in NMDA conductance and, rather than a form of bistability, is due to the voltage-dependent blockade of NMDA receptors. Our models also predict a more pronounced membrane potential bimodality following D1 receptor activation. This work thus establishes reduced yet accurate dopamine-modulated models of MSNs, suitable for use in large-scale models of the striatum. More importantly, these provide a tractable framework for further study of dopamine's effects on computation by individual neurons.


Kidney cytosine methylation changes improve renal function decline estimation in patients with diabetic kidney disease.
Epigenetic changes might provide the biological explanation for the long-lasting impact of metabolic alterations of diabetic kidney disease development. Here we examined cytosine methylation of human kidney tubules using Illumina Infinium 450 K arrays from 91 subjects with and without diabetes and varying degrees of kidney disease using a cross-sectional design. We identify cytosine methylation changes associated with kidney structural damage and build a model for kidney function decline. We find that the methylation levels of 65 probes are associated with the degree of kidney fibrosis at genome wide significance. In total 471 probes improve the model for kidney function decline. Methylation probes associated with kidney damage and functional decline enrich on kidney regulatory regions and associate with gene expression changes, including epidermal growth factor (EGF). Altogether, our work shows that kidney methylation differences can be detected in patients with diabetic kidney disease and improve kidney function decline models indicating that they are potentially functionally important.


Strong mitochondrial DNA support for a Cretaceous origin of modern avian lineages.
Determining an absolute timescale for avian evolutionary history has proven contentious. The two sources of information available, paleontological data and inference from extant molecular genetic sequences (colloquially, 'rocks' and 'clocks'), have appeared irreconcilable; the fossil record supports a Cenozoic origin for most modern lineages, whereas molecular genetic estimates suggest that these same lineages originated deep within the Cretaceous and survived the K-Pg (Cretaceous-Paleogene; formerly Cretaceous-Tertiary or K-T) mass-extinction event. These two sources of data therefore appear to support fundamentally different models of avian evolution. The paradox has been speculated to reflect deficiencies in the fossil record, unrecognized biases in the treatment of genetic data or both. Here we attempt to explore uncertainty and limit bias entering into molecular divergence time estimates through: (i) improved taxon (n = 135) and character (n = 4594 bp mtDNA) sampling; (ii) inclusion of multiple cladistically tested internal fossil calibration points (n = 18); (iii) correction for lineage-specific rate heterogeneity using a variety of methods (n = 5); (iv) accommodation of uncertainty in tree topology; and (v) testing for possible effects of episodic evolution. The various 'relaxed clock' methods all indicate that the major (basal) lineages of modern birds originated deep within the Cretaceous, although temporal intraordinal diversification patterns differ across methods. We find that topological uncertainty had a systematic but minor influence on date estimates for the origins of major clades, and Bayesian analyses assuming fixed topologies deliver similar results to analyses with unconstrained topologies. We also find that, contrary to expectation, rates of substitution are not autocorrelated across the tree in an ancestor-descendent fashion. Finally, we find no signature of episodic molecular evolution related to either speciation events or the K-Pg boundary that could systematically mislead inferences from genetic data. The 'rock-clock' gap has been interpreted by some to be a result of the vagaries of molecular genetic divergence time estimates. However, despite measures to explore different forms of uncertainty in several key parameters, we fail to reconcile molecular genetic divergence time estimates with dates taken from the fossil record; instead, we find strong support for an ancient origin of modern bird lineages, with many extant orders and families arising in the mid-Cretaceous, consistent with previous molecular estimates. Although there is ample room for improvement on both sides of the 'rock-clock' divide (e.g. accounting for 'ghost' lineages in the fossil record and developing more realistic models of rate evolution for molecular genetic sequences), the consistent and conspicuous disagreement between these two sources of data more likely reflects a genuine difference between estimated ages of (i) stem-group origins and (ii) crown-group morphological diversifications, respectively. Further progress on this problem will benefit from greater communication between paleontologists and molecular phylogeneticists in accounting for error in avian lineage age estimates.


Brominated and chlorinated flame retardants in liver of Greenland shark (Somniosus microcephalus).
Polybrominated diphenyl ethers (PBDEs) are known brominated flame retardants that have now been banned or phased out in many parts of the world. As a consequence, interest in the environmental occurrence of non-PBDE flame retardants has increased. In the present study several potential PBDE replacement products together with short chained chlorinated paraffins (SCCPs) were assessed in Greenland sharks accidentally caught in waters around Iceland between 2001 and 2003. Non-PBDE flame retardants detected were pentabromoethylbenzene (PBEB), 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) and 2,3,5,6-tetrabromo-p-xylene (TBX). The concentrations were lower than levels of BDE-47 but similar to other PBDE congeners previously reported in Greenland shark. The median concentrations of SCCPs was 430 ng g(-1) fat, similar to individual PCB congeners previously reported. This is the first report of SCCPs, BTBPE, PBEB and TBX in any shark species globally and confirms the usefulness of the Greenland shark as a screening species for environmental contamination in the Arctic and sub-Arctic environment.


The aromatic fluctuation index (FLU): a new aromaticity index based on electron delocalization.
In this work, the aromatic fluctuation index (FLU) that describes the fluctuation of electronic charge between adjacent atoms in a given ring is introduced as a new aromaticity measure. This new electronic criterion of aromaticity is based on the fact that aromaticity is related to the cyclic delocalized circulation of pi electrons. It is defined not only considering the amount of electron sharing between contiguous atoms, which should be substantial in aromatic molecules, but also taking into account the similarity of electron sharing between adjacent atoms. For a series of rings in 15 planar polycyclic aromatic hydrocarbons, we have found that, in general, FLU is strongly correlated with other widely used indicators of local aromaticity, such as the harmonic-oscillator model of aromaticity, the nucleus independent chemical shift, and the para-delocalization index (PDI). In contrast to PDI, the FLU index can be applied to study the aromaticity of rings with any number of members and it can be used to analyze both the local and global aromatic character of rings and molecules.


[Adverse events and near misses in medical imaging].
In 2000, the Institute of Medicine's report, To Err Is Human: Building a Safer Health System, caught the public attention documenting the magnitude of the medical error problem and the inherent patient safety: medical errors cause between 44,000 and 98,000 deaths annually in the United States. Currently, there is a growing interest in risk management on the medical field, particularly in the management of adverse events. It has been mainly due to the commitment of the World Health Organization, that this field of research has gained increasing the attention it deserves. Medical imaging is one of the high risk fields for the occurrence of errors, especially due to the multiplicity of techniques, the several stakeholders and the complexity of the whole circuit that involves the conduct of studies. Many of the methods used to analyze patient safety were adapted from risk-management techniques in high-risk industries (e.g. chemical, nuclear power and aviation industry). It is recognized that we can learn more from our mistakes than from our successes and the reporting systems in these industries have provided a valuable contribution to error prevention and risk management techniques. At a minimum, adverse events reporting systems can help to identify hazards and risks, providing important information on the system aspects that should be improved. However, the accumulation of potentially relevant data contributes little to healthcare services improvement. It is crucial to apply models to identify the underlying system failures, the root causes, and enhance the sharing of knowledge and experience. In this paper, it is suggested a solution to reduce adverse events, by identifying and eliminating the root causes that are in their source. How the Eindhoven Classification Model was adapted and extended specifically for the Medical Imaging field is also presented. The proposed approach includes the root causes analysis and introduces incomplete information concepts through the use of logical-mathematical operators formally sustained. This model is the basis of the adverse event and near misses reporting and learning system that was developed for Medical Imaging and is implemented in two Portuguese healthcare institutions. The objectives, characteristics and function of this system are presented throughout this article.


Practice-based research in psychiatry.
The authors describe the APA Practice Research Network (PRN), a national research initiative that ultimately will engage 1,000 practicing psychiatrists in collaborative clinical and services research. The PRN is designed to generate practice-relevant information and to inform future service delivery, policy, and financing decisions pertinent to psychiatry. The authors review the relative strengths and limitations of practice-based research compared with other widely used research methods. They also describe the structure of the PRN and its procedures for recruiting network members and for identifying and developing specific network studies. The three primary sources of data for the PRN are 1) the biannual National Survey of Psychiatric Practice, which provides not only a mechanism for randomly recruiting the two-thirds of network participants who are not volunteers but also a baseline for assessing the generalizability of PRN findings; 2) separate biannual studies of psychiatric patients and treatments to characterize the network patient/treatment denominator, which is used to monitor trends in psychiatric practice and plan network studies; and 3) specific studies. Pilot data from the PRN have yielded detailed information on the diagnostic and clinical characteristics of 725 patients and on the treatments provided by network psychiatrists. The APA PRN offers a powerful complement to traditional clinical and health services research approaches. The PRN will help psychiatry improve its ability to meet patients needs in a context of rapidly evolving scientific and clinical progress and legislative and economic forces affecting health care delivery.


WEIRD languages have misled us, too.
The linguistic and cognitive sciences have severely underestimated the degree of linguistic diversity in the world. Part of the reason for this is that we have projected assumptions based on English and familiar languages onto the rest. We focus on some distortions this has introduced, especially in the study of semantics.


Multivariate analysis of 1.5 million people identifies genetic associations with traits related to self-regulation and addiction.
Behaviors and disorders related to self-regulation, such as substance use, antisocial behavior and attention-deficit/hyperactivity disorder, are collectively referred to as externalizing and have shared genetic liability. We applied a multivariate approach that leverages genetic correlations among externalizing traits for genome-wide association analyses. By pooling data from ~1.5 million people, our approach is statistically more powerful than single-trait analyses and identifies more than 500 genetic loci. The loci were enriched for genes expressed in the brain and related to nervous system development. A polygenic score constructed from our results predicts a range of behavioral and medical outcomes that were not part of genome-wide analyses, including traits that until now lacked well-performing polygenic scores, such as opioid use disorder, suicide, HIV infections, criminal convictions and unemployment. Our findings are consistent with the idea that persistent difficulties in self-regulation can be conceptualized as a neurodevelopmental trait with complex and far-reaching social and health correlates.


The Caenorhabditis elegans genes egl-27 and egr-1 are similar to MTA1, a member of a chromatin regulatory complex, and are redundantly required for embryonic patterning.
We show here that two functionally redundant Caenorhabditis elegans genes, egl-27 and egr-1, have a fundamental role in embryonic patterning. When both are inactivated, cells in essentially all regions of the embryo fail to be properly organised. Tissue determination and differentiation are unaffected and many zygotic patterning genes are expressed normally, including HOX genes. However, hlh-8, a target of the HOX gene mab-5, is not expressed. egl-27 and egr-1 are members of a gene family that includes MTA1, a human gene with elevated expression in metastatic carcinomas. MTA1 is a component of a protein complex with histone deacetylase and nucleosome remodelling activities. We propose that EGL-27 and EGR-1 function as part of a chromatin regulatory complex required for the function of regional patterning genes.


3D source localization of interictal spikes in epilepsy patients with MRI lesions.
The present study aims to accurately localize epileptogenic regions which are responsible for epileptic activities in epilepsy patients by means of a new subspace source localization approach, i.e. first principle vectors (FINE), using scalp EEG recordings. Computer simulations were first performed to assess source localization accuracy of FINE in the clinical electrode set-up. The source localization results from FINE were compared with the results from a classic subspace source localization approach, i.e. MUSIC, and their differences were tested statistically using the paired t-test. Other factors influencing the source localization accuracy were assessed statistically by ANOVA. The interictal epileptiform spike data from three adult epilepsy patients with medically intractable partial epilepsy and well-defined symptomatic MRI lesions were then studied using both FINE and MUSIC. The comparison between the electrical sources estimated by the subspace source localization approaches and MRI lesions was made through the coregistration between the EEG recordings and MRI scans. The accuracy of estimations made by FINE and MUSIC was also evaluated and compared by R(2) statistic, which was used to indicate the goodness-of-fit of the estimated sources to the scalp EEG recordings. The three-concentric-spheres head volume conductor model was built for each patient with three spheres of different radii which takes the individual head size and skull thickness into consideration. The results from computer simulations indicate that the improvement of source spatial resolvability and localization accuracy of FINE as compared with MUSIC is significant when simulated sources are closely spaced, deep, or signal-to-noise ratio is low in a clinical electrode set-up. The interictal electrical generators estimated by FINE and MUSIC are in concordance with the patients' structural abnormality, i.e. MRI lesions, in all three patients. The higher R(2) values achieved by FINE than MUSIC indicate that FINE provides a more satisfactory fitting of the scalp potential measurements than MUSIC in all patients. The present results suggest that FINE provides a useful brain source imaging technique, from clinical EEG recordings, for identifying and localizing epileptogenic regions in epilepsy patients with focal partial seizures. The present study may lead to the establishment of a high-resolution source localization technique from scalp-recorded EEGs for aiding presurgical planning in epilepsy patients.


Awareness and knowledge on eye donation among university students.
Four hundred (400) students studying first year Medicine, Dentistry, Laboratory Technology, Pharmacy, Biomedicine and Bioengineering degree courses in the University of Malaya were assessed on their awareness and knowledge on eye donation using an open ended questionnaire. The majority of the students (344, 86%) in this study were aware about eye donation; the awareness was higher in biomedical (77.1%) and medical students (76.7%) compared to the others (55.9%-70.7%). One hundred and eight students (27%) were willing to donate their eyes. Most of the students (376, 94%) did not know about any eye bank in Malaysia. One hundred and sixty (40%) students were aware that whole eye can be removed from the donor and 101 (25.25%) were aware that the cornea can be removed separately. However, only 121 (30.25%) knew that donated eyes were used for corneal grafting. More than half of the students (231, 57.7%) did not know that the donor eye could be stored before transplantation. The results of this study indicate that there is a need to educate the young adults in our society about corneal transplantation so that they can in turn motivate other members of society and their own family members to become eye donors, thus facilitating the availability of donor corneas for corneal transplantation in Malaysia.


Interventions for hyperhidrosis in secondary care: a systematic review and value-of-information analysis.
Hyperhidrosis is uncontrollable excessive sweating that occurs at rest, regardless of temperature. The symptoms of hyperhidrosis can significantly affect quality of life. The management of hyperhidrosis is uncertain and variable. To establish the expected value of undertaking additional research to determine the most effective interventions for the management of refractory primary hyperhidrosis in secondary care. A systematic review and economic model, including a value-of-information (VOI) analysis. Treatments to be prescribed by dermatologists and minor surgical treatments for hyperhidrosis of the hands, feet and axillae were reviewed; as endoscopic thoracic sympathectomy (ETS) is incontestably an end-of-line treatment, it was not reviewed further. Fifteen databases (e.g. CENTRAL, PubMed and PsycINFO), conference proceedings and trial registers were searched from inception to July 2016. Systematic review methods were followed. Pairwise meta-analyses were conducted for comparisons between botulinum toxin (BTX) injections and placebo for axillary hyperhidrosis, but otherwise, owing to evidence limitations, data were synthesised narratively. A decision-analytic model assessed the cost-effectiveness and VOI of five treatments (iontophoresis, medication, BTX, curettage, ETS) in 64 different sequences for axillary hyperhidrosis only. Fifty studies were included in the effectiveness review: 32 randomised controlled trials (RCTs), 17 non-RCTs and one large prospective case series. Most studies were small, rated as having a high risk of bias and poorly reported. The interventions assessed in the review were iontophoresis, BTX, anticholinergic medications, curettage and newer energy-based technologies that damage the sweat gland (e.g. laser, microwave). There is moderate-quality evidence of a large statistically significant effect of BTX on axillary hyperhidrosis symptoms, compared with placebo. There was weak but consistent evidence for iontophoresis for palmar hyperhidrosis. Evidence for other interventions was of low or very low quality. For axillary hyperhidrosis cost-effectiveness results indicated that iontophoresis, BTX, medication, curettage and ETS was the most cost-effective sequence (probability 0.8), with an incremental cost-effectiveness ratio of £9304 per quality-adjusted life-year. Uncertainty associated with study bias was not reflected in the economic results. Patients and clinicians attending an end-of-project workshop were satisfied with the sequence of treatments for axillary hyperhidrosis identified as being cost-effective. All patient advisors considered that the Hyperhidrosis Quality of Life Index was superior to other tools commonly used in hyperhidrosis research for assessing quality of life. The evidence for the clinical effectiveness and safety of second-line treatments for primary hyperhidrosis is limited. This meant that there was insufficient evidence to draw conclusions for most interventions assessed and the cost-effectiveness analysis was restricted to hyperhidrosis of the axilla. Based on anecdotal evidence and inference from evidence for the axillae, participants agreed that a trial of BTX (with anaesthesia) compared with iontophoresis for palmar hyperhidrosis would be most useful. The VOI analysis indicates that further research into the effectiveness of existing medications might be worthwhile, but it is unclear that such trials are of clinical importance. Research that established a robust estimate of the annual incidence of axillary hyperhidrosis in the UK population would reduce the uncertainty in future VOI analyses. This study is registered as PROSPERO CRD42015027803. The National Institute for Health Research Health Technology Assessment programme.


Median nerve: neurophysiological parameters in third trimester of pregnancy.
An examination of neurophysiologic features of median nerve in third trimester of regularly controlled normal risk pregnancies is performed at the Department of Neurophysiology of Primary Health Centre in Tuzla during January / April 2006. Examined group consisted of 40 young females in third trimester of pregnancy, and average age of 25,6 +/- 4,9 years. Control group consisted of young healthy females with average age of 31,1 +/- 4,4 years. Symptoms and signs of carpal tunnel sy. (CST) had 12 patients, but diagnosis is neurophysiologically confirmed in 9 (75%) patients. In group of pregnant females without symptoms 3 (10,7%) patients showed neurophysiologic evidence of CTS. Sensory velocity of right median nerve was significantly lower in pregnant group of patients (p=0,002), but area of sensory potentials on both sides were lower in pregnant group (p<0,0001). Area of CMAP of right median nerve was significantly lower in pregnant group (p=0,0003). Significant differences in CTS group compared with control group were in sensory velocities of median nerve (left median nerve p=0,0007, right p<0,0001), and area of SNAP of both sides (left p<0,0001, right p=0,0001), but area of CMAP right (p=0,0003). In CTS group 7 females had unilateral and 5 had bilateral neurophysiological changes. Our conclusion is that neurophysiological parameters of median nerve in third trimester of pregnancy are changed mainly due to high prevalence of CTS that might disturb quality of life and have psychological and physical implications on future mother. Hence, it is necessary to, continuously, pay enough attention in prevention or treatment of mentioned syndrome in this population group.


If You Make it Free, Will They Come? Using a Physical Activity Accessibility Model to Understand the Use of a Free Children's Recreation Pass.
Children's sedentary lifestyles and low physical activity levels may be countered using population-level interventions. This study examines factors influencing the use of a free community-wide physical activity access pass for grade 5 students (G5AP). A natural experiment with longitudinal data collection. A sample of 881 children completed the 9-month follow-up survey self-reporting where they used the G5AP. Two analyses were conducted: Getis-Ord GI* geographic cluster analysis of the spatial distribution of users, and logistic regression examining the relationship between use and accessibility (informational, economic, and geographic) and mobility options, while accounting for intrapersonal and interpersonal factors. Overall, 44.9% of children used the G5AP with clusters of high use in urban areas and low use in the suburbs. Other factors significantly related to G5AP included gender (girls), informational accessibility (active recruitment), economic accessibility (median household income), geographic accessibility (facilities within 1.6 km of home), and mobility options (access to Boys & Girls Club bus). This study found that a diverse population of children used the G5AP. To continue being successful, community-based physical activity interventions need to ensure that the intervention increases geographic, economic, and informational accessibility and provides mobility options that are available to the target population.


Pro isomerization in MLL1 PHD3-bromo cassette connects H3K4me readout to CyP33 and HDAC-mediated repression.
The MLL1 gene is a frequent target for recurrent chromosomal translocations, resulting in transformation of hematopoietic precursors into leukemia stem cells. Here, we report on structure-function studies that elucidate molecular events in MLL1 binding of histone H3K4me3/2 marks and recruitment of the cyclophilin CyP33. CyP33 contains a PPIase and a RRM domain and regulates MLL1 function through HDAC recruitment. We find that the PPIase domain of CyP33 regulates the conformation of MLL1 through proline isomerization within the PHD3-Bromo linker, thereby disrupting the PHD3-Bromo interface and facilitating binding of the MLL1-PHD3 domain to the CyP33-RRM domain. H3K4me3/2 and CyP33-RRM target different surfaces of MLL1-PHD3 and can bind simultaneously to form a ternary complex. Furthermore, the MLL1-CyP33 interaction is required for repression of HOXA9 and HOXC8 genes in vivo. Our results highlight the role of PHD3-Bromo cassette as a regulatory platform, orchestrating MLL1 binding of H3K4me3/2 marks and cyclophilin-mediated repression through HDAC recruitment.


Language phenotypes in children with sex chromosome trisomies.
Background: Sex chromosome trisomies (47,XXX, 47,XXY and 47,XYY) are known to be a risk factor for language disorder, but typical outcomes are hard to estimate, because many cases are identified only when problems are found. Methods: We recruited children aged 5-16 years with all three types of trisomy, and divided them into a High Bias group, identified in the course of investigations for neurodevelopmental problems, and a Low Bias group, identified via prenatal screening or other medical investigations. Children from a twin sample were used to compare the pattern and severity of language problems: they were subdivided according to parental concerns about language/history of speech-language therapy into a No Concerns group (N = 132) and a Language Concerns group (N = 41). Children were individually assessed on a psychometric battery, and a standardized parent checklist. After excluding children with intellectual disability, autism or hearing problems, psychometric data were available for 31 XXX, 20 XXY and 19 XYY Low Bias cases and 13 XXX, 25 XXY and 32 XYY High Bias cases. Results: Variation within each trisomy group was substantial: within the Low Bias group, overall language scores were depressed relative to normative data, but around one-third had no evidence of problems. There was no effect of trisomy type, and the test profile was similar to the Language Concerns comparison group. The rate of problems was much greater in the High Bias children with trisomies. Conclusions: When advising parents after discovery of a trisomy, it is important to emphasise that, though there is an increased risk of language problems, there is a very wide range of outcomes. Severe language problems are more common in those identified via genetic testing for neurodevelopmental problems but these are not typical of children identified on prenatal screening.


Refining retinoic acid stimulation for osteogenic differentiation of murine adipose-derived adult stromal cells.
Murine adipose-derived adult stromal cells (ADAS) seeded onto appropriate scaffolds and pre-incubated with retinoic acid have been shown to generate in vivo bone rapidly. Prompt resorption ensues, however, as a result of osteoclastogenesis, likely secondary to retinoic acid carryover. In this study, we determined the effects of abbreviated retinoic acid exposure on ADAS osteogenic differentiation. Histological staining and gene expression analysis revealed that longer retinoic acid exposure resulted in better in vitro bone differentiation. However, significant osteogenesis was observed in ADAS after just 15 days of retinoic acid supplementation, suggesting that continual culture with retinoic acid is unnecessary for initiation of the osteogenic program. This was confirmed using ADAS pre-incubated in monolayer with an abbreviated 15 days of retinoic acid exposure before implantation into critical-sized calvarial defects. Similar rates of regeneration were observed between ADAS exposed to for 15 days or for a full 25-day course of retinoic acid before defect repair. Furthermore, by limiting retinoic acid exposure to ADAS in monolayer without scaffold, accelerated bone formation was observed without concomitant osteoclastic resorption. These data suggest that skeletal regeneration may be improved by modulating retinoic acid exposure before implantation, markedly accelerating the repair of bone defects using ADAS.


Expanding Omics Resources for Improvement of Soybean Seed Composition Traits.
Food resources of the modern world are strained due to the increasing population. There is an urgent need for innovative methods and approaches to augment food production. Legume seeds are major resources of human food and animal feed with their unique nutrient compositions including oil, protein, carbohydrates, and other beneficial nutrients. Recent advances in next-generation sequencing (NGS) together with "omics" technologies have considerably strengthened soybean research. The availability of well annotated soybean genome sequence along with hundreds of identified quantitative trait loci (QTL) associated with different seed traits can be used for gene discovery and molecular marker development for breeding applications. Despite the remarkable progress in these technologies, the analysis and mining of existing seed genomics data are still challenging due to the complexity of genetic inheritance, metabolic partitioning, and developmental regulations. Integration of "omics tools" is an effective strategy to discover key regulators of various seed traits. In this review, recent advances in "omics" approaches and their use in soybean seed trait investigations are presented along with the available databases and technological platforms and their applicability in the improvement of soybean. This article also highlights the use of modern breeding approaches, such as genome-wide association studies (GWAS), genomic selection (GS), and marker-assisted recurrent selection (MARS) for developing superior cultivars. A catalog of available important resources for major seed composition traits, such as seed oil, protein, carbohydrates, and yield traits are provided to improve the knowledge base and future utilization of this information in the soybean crop improvement programs.


Biomechanical effect of patch graft for large rotator cuff tears: a cadaver study.
It is not possible for some rotator cuff tears to be repaired because of a large defect associated with muscle retraction. The purpose of the current study was to investigate the use of a synthetic patch graft to restore abduction force transmission in the glenohumeral joint with a rotator cuff defect. Shoulders from cadavers (n = 10) were fixed in the hanging arm and in neutral rotation, and loading was applied to the rotator cuff tendons and middle deltoid. After a simulated supraspinatus tendon defect and retraction, a patch graft was inserted into the defect and the effects of reattachment to the greater tuberosity, narrowing of the defect by using a smaller graft, and anterior graft attachment (rotator interval tissue versus subscapularis) were investigated. Abduction torque generation was measured and normalized to the intact condition. Compared with torque generation after creation of a supraspinatus defect (61% of normal torque), abduction torque increased with a graft between the infraspinatus and either the rotator interval (68% of normal) or subscapularis (80% of normal). The optimum grafting technique for abduction torque restoration occurred with a reduced size patch connected anteriorly to the subscapularis and sutured to the greater tuberosity (107% of normal). The patch graft acts to redirect force transmission, thereby providing a potential treatment option for otherwise irreparable defects. These same principles can be applied when tendon transfers are used to reconstruct large or massive cuff tears.


Cryogenic colocalization microscopy for nanometer-distance measurements.
The main limiting factor in spatial resolution of localization microscopy is the number of detected photons. Recently we showed that cryogenic measurements improve the photostability of fluorophores, giving access to Angstrom precision in localization of single molecules. Here, we extend this method to colocalize two fluorophores attached to well-defined positions of a double-stranded DNA. By measuring the separations of the fluorophore pairs prepared at different design positions, we verify the feasibility of cryogenic distance measurement with sub-nanometer accuracy. We discuss the important challenges of our method as well as its potential for further improvement and various applications.


Factors Affecting Treatment with Life-Saving Interventions, Computed Tomography Scans and Specialist Consultations.
Background: Emergency treatments determined by emergency physicians may affect mortality and patient satisfaction. This paper attempts to examine the impact of patient characteristics, health status, the accredited level of hospitals, and triaged levels on the following emergency treatments: immediate life-saving interventions (LSIs), computed tomography (CT) scans, and specialist consultations (SCs). Methods: A multivariate logistic regression model was employed to analyze the impact of patient characteristics, including sex, age, income and the urbanization degree of the patient's residence; patient health status, including records of hospitalization and the number of instances of ambulatory care in the previous year; the Charlson Comorbidity Index (CCI) score; the accredited level of hospitals; and the triaged level of emergency treatments. Results: All the patient characteristics were found to impact receiving LSI, CT and SC, except for income. Furthermore, a better health status was associated with a decreased probability of receiving LSI, CT and SC, but the number of instances of ambulatory care was not found to have a significant impact on receiving CT or SC. This study also found no evidence to support impact of CCI on SC. Hospitals with higher accredited levels were associated with a greater chance of patients receiving emergency treatments of LSI, CT and SC. A higher assigned severity (lower triaged level) led to an increased probability of receiving CT and SC. In terms of LSI, patients assigned to level 4 were found to have a lower chance of treatment than those assigned to level 5. Conclusions: This study found that several patient characteristics, patient health status, the accredited level of medical institutions and the triaged level, were associated with a higher likelihood of receiving emergency treatments. This study suggests that the inequality of medical resources among medical institutions with different accredited levels may yield a crowding-out effect.


The effect of osteoclastic activity on tendon-to-bone healing: an experimental study in rabbits.
Healing of a tendon graft in a bone tunnel depends on bone ingrowth into the interface between tendon and bone. Excessive osteoclastic activity may contribute to bone resorption, tunnel widening, and impaired healing. We hypothesized that inhibition of osteoclastic activity by osteoprotegerin (OPG) would increase bone formation around a tendon graft in anterior cruciate ligament reconstruction in a rabbit model, while increased osteoclastic activity due to the application of receptor activator of nuclear factor-kappa B ligand (RANKL) would impair bone ingrowth. Sixty skeletally mature, male New Zealand White rabbits underwent bilateral anterior cruciate ligament reconstruction. OPG (100 microg per tunnel) or RANKL (10 microg per tunnel) was delivered to the tendon-bone interface with use of a synthetic calcium phosphate carrier vehicle. Twenty animals were killed at two, four, and eight weeks after surgery. Two rabbits from each group were prepared for histological evaluation, and the other rabbits were used for biomechanical testing. A significantly greater amount of bone surrounded the tendon at the healing tendon-bone interface in the OPG-treated limbs compared with the controls and the RANKL-treated limbs at all time-points (p < 0.05). There were significantly fewer osteoclasts in the OPG-treated limbs compared with the controls and the RANKL-treated limbs (p < 0.05). The average tunnel area in the OPG group was significantly smaller than that in the RANKL group (p = 0.003 at two weeks and p = 0.004 at four weeks). The femur-anterior cruciate ligament-tibia complex of the OPG-treated limbs had significantly increased stiffness compared with RANKL-treated limbs at eight weeks (p = 0.04). Osteoprotegerin significantly improves bone formation around the grafted tendon and improves the stiffness at the healing tendon-bone junction in a rabbit model.


Mother-to-Infant Bonding in Women with Postpartum Psychosis and Severe Postpartum Depression: A Clinical Cohort Study.
Mother-to-infant bonding is important for long-term child development. The aim of this study was to investigate bonding in women admitted to a Mother and Baby Unit with postpartum depression (PD, n = 64) and postpartum psychosis (PP, n = 91). Participants completed the Postpartum Bonding Questionnaire (PBQ), the Edinburgh Postnatal Depression Scale (EPDS) and the Young Mania Rating Scale (YMRS) weekly during admission. At admission, 57.1% of women with PD had impaired bonding, compared to only 17.6% of women with PP (p-value < 0.001). At discharge, only 18.2% of women with PD and 5.9% of women with PP still experienced impaired bonding (p-value = 0.02). There was a strong association between decrease of depressive and manic symptoms and improved bonding over an eight-week admission period. In a small group of women (5.7%) impaired bonding persisted despite being in remission of their psychiatric disorder. The results from our study show that impaired bonding is a more present and evidently severe problem in postpartum depression but not so much in postpartum psychosis. Treatment of depressive symptoms will improve bonding in almost all women, but clinicians should assess if impaired bonding is still present after remission because for a small group special care and treatment focused on bonding might be required.


In vivo genotoxicity of treated water containing the cylindrospermopsin-producer Cylindrospermopsis raciborskii.
Cylindrospermopsin (CYN) is an alkaloid commonly produced by some cyanobacteria that has been implicated in outbreaks of human illness. The aim of this study was to investigate the genotoxicity of Cylindrospermopsis raciborskii cellular content (including CYN) and its byproducts resulting from chlorination during water treatment. DNA damage in blood and liver cells was analysed by the comet assay and micronucleus test (MN). Mice were injected intraperitoneally with the following treatments: (a) physiological saline, (b) treated water, (c) treated water plus C. raciborskii extract (CYN producer strain, CYPO-011 K), (d) C. raciborskii extract (CYN producer strain, CYPO-011 K), (e) C. raciborskii extract (CYN non producer strain), and (f) treated water plus C. raciborskii extract (CYN non producer strain) extract. After 48 h, samples were taken to perform tests (blood and liver cells to the comet assay and bone marrow to MN test). The CYPO-011 K had a genotoxic and mutagenic effects on liver and bone marrow cells. The group that received chlorine-treated water plus CYPO-011 K also exhibited genotoxic effects in the liver, as well as in the blood, and a mutagenic effect in blood marrow cells. The results emphasise the need of improving CYN monitoring in waters bodies in order to reduce the risk of human exposure.


Optimal periodic perfusion strategy for robust long-term microfluidic cell culture.
Long-term cell culture in microfluidic devices is an essential prerequisite for "on a chip" biological and physiological based studies. We investigated how medium delivery, from continuous to periodic perfusion, affects long-term cell cultures in a microfluidic platform. Computational simulations suggested that different delivery strategies result in different temporal profiles of accumulation and washing out of endogenous (EnF) and exogenous (ExF) factors, respectively. Thus, cultures exposed to the same overall amount of medium with different temporal profiles were analysed in terms of homogeneity, cell morphology and phenotype. Murine and human cell lines (C2C12 and HFF) and mouse embryonic stem cells (mESC) were cultured in microfluidic channels. An ad hoc experimental setup was developed to perform continuous and periodic medium delivery into the chip, tuning the flow rate, the perfusion time, and the interval of perfusion while using the same amount of medium volume. Periodic medium delivery with a short perfusion pulse ensured cell homogeneity compared to standard cell culture. Conversely, a continuous flow resulted in cell heterogeneity, with abnormal morphology and vesiculation. Only dramatic and unfeasible increasing of perfused medium volume in the continuous configuration could rescue normal cell behaviour. Consistent results were obtained for C2C12 and HFF. In order to extend these results to highly sensitive cells, mESC were cultured for 6 days in the microfluidic channels. Our analysis demonstrates that a periodic medium delivery with fast pulses (with a frequency of 4 times per day) resulted in a homogeneous cell culture in terms of cell viability, colony morphology and maintenance of pluripotency markers. According to experimental observations, the computational model provided a rational description of the perfusion strategies and of how they deeply shape the cell microenvironment in microfluidic cell cultures. These results provide new insight to define optimal strategies for homogeneous and robust long-term cell culture in microfluidic systems, an essential prerequisite for lab on chip cell-based applications.


Neutropenia due to copper deficiency in total parenteral nutrition.
Copper (Cu) deficiency has been reported both in pediatric and adult patients on total parenteral nutrition (TPN). Manifestations of Cu deficiency are usually hematologic in the adult. A 56-yr-old patient with a history of subtotal gastrectomy underwent massive small bowel resection and partial colectomy in 1977. TPN was initiated. Intravenous (iv) trace mineral supplements were not available in 1977 at our institution. By June 1978, the patient manifested neutropenia with a white blood cell (WBC) count of 2000/mm3 and 39% neutrophils or a total neutrophil count (TNC) of 780/mm3. The serum Cu level at that time was 5 micrograms/dl (normal, 70-140), and the serum zinc (Zn) level was low, although within normal limits. Oral supplementation with Cu and Zn tablets proved ineffective. Twice weekly infusions of fresh frozen plasma were begun in April 1979, and intravenous chromium supplementation started in September 1979, but neither the WBC count nor the TNC improved. Intravenous Cu and Zn were approved in October 1979, and were added to the TPN formula. Within 2 weeks, the WBC count rose to 6300/mm3, and the neutrophils increased to 83% with a TNC of 5229. After 12 weeks, the serum Cu level rose to 80 micrograms/dl. The increase in WBC count and percentage of neutrophils can be attributed to the addition of Cu to the TPN infusate.


Managing potential health risks from electric powerlines: a decision analysis caught in controversy.
Over the past 20 years, several epidemiological studies have found an association between exposure to electromagnetic fields (EMFs) and health effects, including childhood leukemia and adult brain cancer. However, experts strongly disagree about whether this association is causal and, if so, how strong it is. In this article, we examine several alternatives to reduce EMFs from sources of the California power grid, including undergrounding distribution and transmission lines and reconfiguring or rephasing lines. The alternatives were evaluated in terms of the potential health risk reduction, cost, impacts on service reliability, property values, and many other consequences. Because of the uncertainty about an EMF-health link, the main effort was to determine the sensitivity of the decisions to the probability and seriousness of an EMF hazard. User-friendly computer models were developed to allow stakeholders to change the model assumptions and parameters to analyze the impacts of their own assumptions and estimates on the decision. The analysis clearly demonstrated that only four of the many concerns raised by the stakeholders could make a difference in the decision: health risks, costs, service reliability, and property values. Whether undergrounding, moderate alternatives for EMF reduction, or no change was the best decision depended on a few key factors, including the probability that EMF exposure is a hazard, the severity of this hazard, how the EMF reduction measures are financed, and the impacts on property values. While the analysis did not resolve the EMF issues, it showed that even in the most controversial settings, a little analysis goes a long way to clarifying the issues and to focus the debate.


Sulphur and skin: from Satan to Saddam!
Since the dawn of time, Beelzebub has been showering fire and brimstone (sulphur) on tortured souls, but the cutaneous effects of this have been poorly described. Sulphur has also been used for centuries as a treatment for many skin conditions, such as fungal infections, scabies, psoriasis, eczema and acne. It has also been used extensively in cosmetic preparations and by cosmetic dermatologists treating conditions such as seborrhoeic eczema. Many natural bathing spas have high levels of sulphur; such balneology has been advocated by medical and cosmetic dermatologists as an effective treatment for cutaneous disorders for more than 500 years. Sulphur was often the active agent in many of the so-called 'patent medicines' that became popular in the mid-nineteenth century. Time has not withered medical practitioners' enthusiasm for sulphur. There are various reports in the medical literature of its current use. However sulphur treatment is not without its risks; a sulphur spring dermatitis has been described from a spa bath in Taiwan. With the satanic threat of bio-terrorism, some dermatologists may be treating the effects of contact with sulphur mustard all too soon.


Microbial signatures of oral dysbiosis, periodontitis and edentulism revealed by Gene Meter methodology.
Conceptual models suggest that certain microorganisms (e.g., the "red" complex) are indicative of a specific disease state (e.g., periodontitis); however, recent studies have questioned the validity of these models. Here, the abundances of 500+ microbial species were determined in 16 patients with clinical signs of one of the following oral conditions: periodontitis, established caries, edentulism, and oral health. Our goal was to determine if the abundances of certain microorganisms reflect dysbiosis or a specific clinical condition that could be used as a 'signature' for dental research. Microbial abundances were determined by the analysis of 138,718 calibrated probes using Gene Meter methodology. Each 16S rRNA gene was targeted by an average of 194 unique probes (n=25nt). The calibration involved diluting pooled gene target samples, hybridizing each dilution to a DNA microarray, and fitting the probe intensities to adsorption models. The fit of the model to the experimental data was used to assess individual and aggregate probe behavior; good fits (R2>0.90) were retained for back-calculating microbial abundances from patient samples. The abundance of a gene was determined from the median of all calibrated individual probes or from the calibrated abundance of all aggregated probes. With the exception of genes with low abundances (<2 arbitrary units), the abundances determined by the different calibrations were highly correlated (r~1.0). Seventeen genera were classified as 'signatures of dysbiosis' because they had significantly higher abundances in patients with periodontitis and edentulism when contrasted with health. Similarly, 13 genera were classified as 'signatures of periodontitis', and 14 genera were classified as 'signatures of edentulism'. The signatures could be used, individually or in combination, to assess the clinical status of a patient (e.g., evaluating treatments such as antibiotic therapies). Comparisons of the same patient samples revealed high false negatives (45%) for next-generation-sequencing results and low false positives (7%) for Gene Meter results.


Incidence of Diabetes in Colorectal Cancer Survivors.
A higher risk of colorectal cancer (CRC) in patients with diabetes has been well documented. However, little is known regarding diabetes incidence in CRC survivors. This may have substantial impact on CRC survivorship care as well as enhancing the understanding of the interplay between the two diseases. We explored whether the incidence of diabetes was higher among patients with CRC than matched control subjects. Using population-based data from Ontario, Canada, we generated a dataset comprising 39 707 incident CRC cases and 198 535 age- and sex-matched control subjects (1:5) dating from April 2002 to March 2010. We used cause-specific hazard models to estimate the hazard ratios (HRs) for diabetes overall and in subgroups stratified by receipt of systemic chemotherapy, diagnosis of metastatic disease, and site of cancer. During a mean follow-up of 4.81 years, the association between CRC and diabetes varied: The rate of developing diabetes was 53% higher among CRC patients compared with control subjects in the first year postdiagnosis (HR = 1.53, 95% confidence interval [CI] = 1.42 to 1.64) and remained increased by 19% in the fifth year postdiagnosis (HR = 1.19, 95% CI = 1.05 to 1.35). Findings were similar in subgroups of patients who had colon cancer, received systemic chemotherapy, or had no evidence of metastasis. We found that CRC patients were statistically significantly more likely to develop subsequent diabetes than persons without CRC for up to five years after the diagnosis. Our study suggests that active screening and counseling regarding modifiable risk factors may be warranted in this high-risk group.


Surface-Tuned and Metal-Ion-Responsive Supramolecular Gels Based on Nucleolipids.
Supramolecular synthons based on nucleic acid components, nucleobases and nucleosides, and their derivatives have been highly useful in constructing wide varieties of nanoarchitectures. While most of the design strategies have focused on developing biocompatible delivery vehicles, the potential of nucleoside hybrids in assembling smart materials with tunable and sensing properties, though challenging, is gaining significant attention. Here, we describe the development of novel functional materials with surface tunability and metal-ion responsiveness by using simple nucleolipid supramolecular synthons derived by attaching various fatty acids to the 3'-O or 3',5'-O positions of the sugar residue of thymidine nucleoside. 3',5'-O-Difatty acid-substituted thymidines formed typical organogels in pure organic solvents, whereas, 3'-O-monofatty acid-substituted thymidine nucleolipids formed water-induced gels. A detailed morphological and structural analysis using microscopy, single-crystal and powder X-ray diffraction, and NMR techniques clearly revealed the molecular interactions invoked by nucleobase, sugar, fatty acid chain, and water in setting up the path for hierarchical self-assembly and gelation of thymidine nucleolipids. Interestingly, the surface property of the xerogel film fabricated using 3'-O-monosubstituted nucleolipid gels could be switched from highly hydrophobic to hydrophilic and vice versa depending on the nature of the organic solvent-water mixture used in the gelation process. On the contrary, the gelation process of disubstituted thymidine nucleolipids was highly sensitive to the presence of Hg2+ ions as the metal ion formed a T-Hg-T base pair, thereby disrupting the H-bonding interactions that favored the gelation. Taken together, straightforward synthesis and modification-dependent gelation behavior, surface tunability, and metal-ion responsiveness underscore the potential of these supramolecular nucleolipid synthons in constructing novel functional materials.


Household and food shopping environments: do they play a role in socioeconomic inequalities in fruit and vegetable consumption? A multilevel study among Dutch adults.
Fruit and vegetables are protective of a number of chronic diseases; however, their intakes have been shown to vary by socioeconomic position (SEP). Household and food shopping environmental factors are thought to contribute to these differences. To determine whether household and food shopping environmental factors are associated with fruit and vegetable (FV) intakes, and contribute to socioeconomic inequalities in FV consumption. Cross-sectional data were obtained by a postal questionnaire among 4333 adults (23-85 years) living in 168 neighbourhoods in the south-eastern Netherlands. Participants agreed/disagreed with a number of statements about the characteristics of their household and food shopping environments, including access, prices and quality. Education was used to characterise socioeconomic position (SEP). Main outcome measures were whether or not participants consumed fruit or vegetables on a daily basis. Multilevel logistic regression models examined between-area variance in FV consumption and associations between characteristics of the household and food shopping environments and FV consumption. Only a few household and food shopping environmental factors were significantly associated with fruit and vegetable consumption, and their prevalence was low. Participants who perceived FV to be expensive were more likely to consume them. There were significant socioeconomic inequalities in fruit and vegetable consumption (ORs of not consuming fruit and vegetables were 4.26 and 5.47 among the lowest-educated groups for fruit and vegetables, respectively); however, these were not explained by any household or food shopping environmental factors. Improving access to FV in the household and food shopping environments will only make a small contribution to improving population consumption levels, and may only have a limited effect in reducing socioeconomic inequalities in their consumption.


Characterization of HIV-1 neutralization escape mutants.
Infection by molecularly cloned HIV-1, in the presence of a high-titre neutralizing monoclonal antibody (MAb), resulted in the selection of plaques in MT4 cells releasing HIV resistant to neutralization by the same MAb. The epitope recognized by the MAb was mapped to the V3 neutralization epitope at amino acids 305-321. The HIV-1 variants showed a reduced binding capacity for the selecting MAb as determined by immunofluorescence. Polymerase chain reaction (PCR) amplification of complementary DNA derived from viral RNA, cloning and sequencing identified a base pair (bp) change C----G at position 6663 in variant 110.5/1, predicting a change at amino acid 308 Arg----Gly. No other changes in the epitope were observed by sequencing three other variants. Differential hybridization of PCR amplified viral RNA and DNA, with oligonucleotides specific for the observed bp change or the 'wild type' sequence, indicated that the variants 110.5/1 and 110.5/7 were genotypically mixed for 308Gly/Arg. Subsequent screening of biologically 'recloned' variants 110.5/1 and 110.5/7 identified two subclones homozygous for the 308Gly change. The Arg----Gly change appears to affect the binding of the antibody to the epitope, since the linear peptide substituting 308Gly for 'wild type' 308Arg was 100 times less potent in blocking the neutralization of parental HIV. Amino-acid residue 308 thus appears to be crucial for antibody binding to the epitope. In addition, mutations distant from the monoclonal antibody binding site may also affect neutralization by antibodies recognizing the V3 loop.


Genomic variation in Plasmodium vivax malaria reveals regions under selective pressure.
Although Plasmodium vivax contributes to almost half of all malaria cases outside Africa, it has been relatively neglected compared to the more deadly P. falciparum. It is known that P. vivax populations possess high genetic diversity, differing geographically potentially due to different vector species, host genetics and environmental factors. We analysed the high-quality genomic data for 46 P. vivax isolates spanning 10 countries across 4 continents. Using population genetic methods we identified hotspots of selection pressure, including the previously reported MRP1 and DHPS genes, both putative drug resistance loci. Extra copies and deletions in the promoter region of another drug resistance candidate, MDR1 gene, and duplications in the Duffy binding protein gene (PvDBP) potentially involved in erythrocyte invasion, were also identified. For surveillance applications, continental-informative markers were found in putative drug resistance loci, and we show that organellar polymorphisms could classify P. vivax populations across continents and differentiate between Plasmodia spp. This study has shown that genomic diversity that lies within and between P. vivax populations can be used to elucidate potential drug resistance and invasion mechanisms, as well as facilitate the molecular barcoding of the parasite for surveillance applications.


Comparison of Rockall and Blatchford scores to assess outcome of patients with bleeding peptic ulcers after endoscopic therapy.
To determine the accuracy of Rockall and Blatchford scores for predicting outcome after endoscopic treatment in two groups of patients with bleeding peptic ulcers: those who initially presented with upper gastrointestinal (UGI) bleeding (Group A) and those who developed UGI bleeding during hospital treatment for another condition (Group B). A total of 593 patients who had had endoscopic treatment for bleeding peptic ulcers from January 2009 to July 2010 were divided into Groups A and B. Endoscopic therapy including monotherapy (thermal therapy or hemoclipping) and combination therapy was applied. The Blatchford and complete Rockall scores for the two subgroups were calculated. Predictive statistics for the use of the two scoring systems were then compared for Groups A and B. Thirty-day re-bleeding and mortality rates increased with increased Rockall and Blatchford scores. Rockall scores were more accurate than the Blatchford scores for predicting mortality. However, neither the Rockall nor the Blatchford score could accurately predict recurrence of bleeding. When the results in Group B and Group A subgroups were compared, the average Rockall score for Group A was lower than that for Group B (5.6 vs. 6.3, p < 0.001). In high-risk patients with peptic ulcer bleeding, the Rockall score can better predict 30-day mortality than can the Blatchford score; this was particularly true for Group B patients.


T cell receptor usage and fine specificity of human immunodeficiency virus 1-specific cytotoxic T lymphocyte clones: analysis of quasispecies recognition reveals a dominant response directed against a minor in vivo variant.
Numerous virus-specific, class I-restricted cytotoxic T lymphocyte (CTL) epitopes have been identified, yet little information is available regarding the specificity of the CTL response in persons of the same human histocompatibility leukocyte antigen (HLA) type. In this study, the human immunodeficiency virus (HIV) 1 envelope-specific CTL response was evaluated in five HLA-B14-positive persons. CTL responses specific for a previously described nine-amino acid epitope in gp41 (aa 584-592, ERYLKDQQL) could be identified in all subjects, and CTL clones specific for this epitope could be isolated from four persons. Despite heterogeneous T cell receptor usage, the fine specificity of the clones was similar, as defined by recognition of alanine-substituted peptides as well as peptides representing natural HIV-1 sequence variants. Correlation with in vivo virus sequences revealed that the dominant species in two of the subjects represented poorly recognized variants, with a K-->Q substitution at amino acid 588, whereas no variants were observed in the other two subjects. Although clonal type-specific responses to these dominant variants could be identified, the magnitude of these responses remained small, and the dominant CTL response was directed at the minor in vivo variant. These studies indicate that despite similar epitope-specific immunologic pressure in persons of the same HLA type, the in vivo quasispecies may differ, and that the major in vivo immune response to a given CTL epitope can be directed at a minor variant.


Vibrio2009: the third international conference on the biology of Vibrios.
Vibrio2009, the third international conference on the biology of Vibrios, was held in Rio de Janeiro, Brazil, in November 2009. This conference united researchers studying various aspects of pathogenesis, symbiosis and environmental persistence of this diverse group of marine bacteria. Through many of the presentations, it became apparent how horizontal gene transfer and genetic flexibility has driven the incredible diversity of these microbes. Interestingly, unifying themes of behaviour could be seen in the interaction(s) of Vibrios with other organisms, such as with other bacteria, corals, invertebrates and humans. Presentations illuminated the idea that the path towards symbiosis is not that different from the path towards disease, and that alterations in environmental conditions, such as climate change, can tip the balance and change the Vibrio interactions from benign to pathogenic.


More than meets the eye: The role of sensory dimensions in psychedelic brain dynamics, experience, and therapeutics.
Psychedelics are undergoing a major resurgence of scientific and clinical interest. While multiple theories and frameworks have been proposed, there is yet no universal agreement on the mechanisms underlying the complex effects of psychedelics on subjective experience and brain dynamics, nor their therapeutic benefits. Despite being prominent in psychedelic phenomenology and distinct from those elicited by other classes of hallucinogens, the effects of psychedelics on low-level sensory - particularly visual - dimensions of experience, and corresponding brain dynamics, have often been disregarded by contemporary research as 'epiphenomenal byproducts'. Here, we review available evidence from neuroimaging, pharmacology, questionnaires, and clinical studies; we propose extensions to existing models, provide testable hypotheses for the potential therapeutic roles of psychedelic-induced visual hallucinations, and simulations of visual phenomena relying on low-level cortical dynamics. In sum, we show that psychedelic-induced alterations in low-level sensory dimensions 1) are unlikely to be entirely causally reconducible to high-level alterations, but rather co-occur with them in a dialogical interplay, and 2) are likely to play a causally relevant role in determining high-level alterations and therapeutic outcomes. We conclude that reevaluating the currently underappreciated role of sensory dimensions in psychedelic states will be highly valuable for neuroscience and clinical practice, and that integrating low-level and domain-specific aspects of psychedelic effects into existing nonspecific models is a necessary step to further understand how these substances effect both acute and long-term change in the human brain.


Actin polymerization and intracellular solvent flow in cell surface blebbing.
The cortical actin gel of eukaryotic cells is postulated to control cell surface activity. One type of protrusion that may offer clues to this regulation are the spherical aneurysms of the surface membrane known as blebs. Blebs occur normally in cells during spreading and alternate with other protrusions, such as ruffles, suggesting similar protrusive machinery is involved. We recently reported that human melanoma cell lines deficient in the actin filament cross-linking protein, ABP-280, show prolonged blebbing, thus allowing close study of blebs and their dynamics. Blebs expand at different rates of volume increase that directly predict the final size achieved by each bleb. These rates decrease as the F-actin concentration of the cells increase over time after plating on a surface, but do so at lower concentrations in ABP-280 expressing cells. Fluorescently labeled actin and phalloidin injections of blebbing cells indicate that a polymerized actin structure is not present initially, but appears later and is responsible for stopping further bleb expansion. Therefore, it is postulated that blebs occur when the fluid-driven expansion of the cell membrane is sufficiently rapid to initially outpace the local rate of actin polymerization. In this model, the rate of intracellular solvent flow driving this expansion decreases as cortical gelation is achieved, whether by factors such as ABP-280, or by concentrated actin polymers alone, thereby leading to decreased size and occurrence of blebs. Since the forces driving bleb extension would always be present in a cell, this process may influence other cell protrusions as well.


Variability in physical constant parameter values from standard data sources and the implication of this variability for risk analysis.
Physical property values are used in environmental risk assessments to estimate media and risk-based concentrations. However, considerable variability has recently been reported with such values. To evaluate potential variability in physical parameter values supporting a variety of regulatory programs, eight data sources were chosen for evaluation, and chemicals appearing in at least four sources were selected. There were 755 chemicals chosen. In addition, chemicals in seven environmentally important subgroups were also identified for evaluation. Nine parameters were selected for analysis-molecular weight (MolWt), melting point (MeltPt), boiling point (BoilPt), vapor pressure (VP), water solubility (AqSOL), Henry's law constant (HLC), octanol-water partition coefficient (Kow), and diffusion coefficients in air (Dair) and water (Dwater). Results show that while 71% of constituents had equal MolWts across data sources, <3% of the constituents had equivalent parameter values across data sources for AqSOL, VP, or HLC. Considerable dissimilarity between certain sources was also observed. Furthermore, measures of dispersion showed considerable variation in data sets for Kow, VP, AqSOL, and HLC compared to measures for MolWt, MeltPt, BoilPt, or Dwater. The magnitude of the observed variability was also noteworthy. For example, the 95th percentile ratio of maximum/minimum parameter values ranged from 1.0 for MolWt to well over 1.0E + 06 for VP, and HLC. Risk and exposure metrics also varied by similar magnitudes. Results with environmentally important subgroups were similar. These results show that there is considerable variability in physical parameter values from standard sources, and that the observed variability could affect potential risk estimates and perhaps risk management decisions.


Is there a correlation between 18F-FDG-PET standardized uptake value, T-classification, histological grading and the anatomic subsites in newly diagnosed squamous cell carcinoma of the head and neck?
(18)F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET)/CT imaging of squamous cell carcinoma of the head and neck (HNSCC) renders the possibility to study metabolic tumor activity by measuring FDG-uptake expressed as maximum standardized uptake value (SUV(max)). A correlation between SUV(max) and several factors including T-classification, histological tumor differentiation or different anatomic subsites is of potential interest in HNSCC. The aim of this study was to evaluate how metabolic tumor activity derived from FDG-PET correlates with prognostic clinical and pathological parameters including these factors. 262 patients with HNSCC undergoing PET/CT for initial staging were assessed separately for a potential correlation between SUV(max) and T-classification, histological grading, and anatomical subsites of the primary tumor. Nonparametric testing showed a significant correlation between SUV(max) and T-classification (P < 0.001). On the contrary, no statistically significant correlation was found between SUV(max) and histological tumor grading. Furthermore, no statistical significant correlation between the different anatomical subsites and SUV(max) were found. There was no significant correlation of SUV(max) and tumor grading after adjustment for T-stage and anatomical localization of the tumor, neither. Metabolic tumor activity correlates with T-stage of HNSCC. However, histological tumor grading does not correlate with SUV(max). The role of primary tumor SUV(max) as a predictor of outcome or survival remains unclear. Clinicians should therefore exercise caution in attributing any clinical importance to SUV(max) obtained from a single PET/CT exam.


An Examination of the Anti-Cancer Properties of Plant Cannabinoids in Preclinical Models of Mesothelioma.
Mesothelioma is an aggressive cancer with limited treatment options and a poor prognosis. Phytocannabinoids possess anti-tumour and palliative properties in multiple cancers, however their effects in mesothelioma are unknown. We investigated the anti-cancer effects and potential mechanisms of action for several phytocannabinoids in mesothelioma cell lines. A panel of 13 phytocannabinoids inhibited growth of human (MSTO and H2452) and rat (II-45) mesothelioma cells in vitro, and cannabidiol (CBD) and cannabigerol (CBG) were the most potent compounds. Treatment with CBD or CBG resulted in G0/G1 arrest, delayed entry into S phase and induced apoptosis. CBD and CBG also significantly reduced mesothelioma cell migration and invasion. These effects were supported by changes in the expression of genes associated with the cell cycle, proliferation, and cell movement following CBD or CBG treatment. Gene expression levels of CNR1, GPR55, and 5HT1A also increased with CBD or CBG treatment. However, treatment with CBD or CBG in a syngeneic orthotopic rat mesothelioma model was unable to increase survival. Our data show that cannabinoids have anti-cancer effects on mesothelioma cells in vitro and alternatives of drug delivery may be needed to enhance their effects in vivo.


Venous thromboembolism and varicose veins share familial susceptibility: a nationwide family study in Sweden.
Varicose veins (VVs) have been associated with venous thromboembolism (VTE), but whether these diseases share familial susceptibility has not been determined. This nationwide study aimed to determine whether VTE shares familial susceptibility with VVs. Swedish Multigeneration Register data for persons aged 0 to 76 years during the period 1964-2008 were linked to the Swedish Inpatient and Outpatient Registers. Familial risks (standardized incidence ratios [SIRs]) of VTE and VVs were examined in 2 ways (ie, bidirectionally): risk of VTE in subjects whose siblings had been diagnosed with VVs and risk of VVs in persons whose siblings had been diagnosed with VTE. The analyses were repeated for spouses to determine the importance of shared adult family environment. In total, 96 810 siblings had VVs and 87 564 had VTE. An increased risk of VTE was observed in persons whose siblings had VVs (SIR 1.30, 95% CI 1.26 to 1.33), whereas persons whose siblings had VTE had an increased risk of VVs (SIR 1.30, 95% CI 1.27 to 1.34). If 2 or more siblings were affected by VTE, the risk for VVs was 1.70 (95% CI 1.53 to 1.88). Conversely, if 2 or more siblings were affected by VVs, the risk for VTE was 1.52 (95% CI 1.38 to 1.67). In spouses of VTE patients, a minor increased risk of VVs was observed (SIR 1.05 for husbands, SIR 1.06 for wives). The risk of VTE in spouses of VV patients was similarly small (SIR 1.01 for husbands, SIR 1.05 for wives). VVs and VTE share familial susceptibility. This novel finding suggests the existence of shared familial and possibly genetic factors.


Association Between Diabetes and Cause-Specific Mortality in Rural and Urban Areas of China.
In China, diabetes prevalence has increased substantially in recent decades, but there are no reliable estimates of the excess mortality currently associated with diabetes. To assess the proportional excess mortality associated with diabetes and estimate the diabetes-related absolute excess mortality in rural and urban areas of China. A 7-year nationwide prospective study of 512 869 adults aged 30 to 79 years from 10 (5 rural and 5 urban) regions in China, who were recruited between June 2004 and July 2008 and were followed up until January 2014. Diabetes (previously diagnosed or detected by screening) recorded at baseline. All-cause and cause-specific mortality, collected through established death registries. Cox regression was used to estimate adjusted mortality rate ratio (RR) comparing individuals with diabetes vs those without diabetes at baseline. Among the 512 869 participants, the mean (SD) age was 51.5 (10.7) years, 59% (n = 302 618) were women, and 5.9% (n = 30 280) had diabetes (4.1% in rural areas, 8.1% in urban areas, 5.8% of men, 6.1% of women, 3.1% had been previously diagnosed, and 2.8% were detected by screening). During 3.64 million person-years of follow-up, there were 24 909 deaths, including 3384 among individuals with diabetes. Compared with adults without diabetes, individuals with diabetes had a significantly increased risk of all-cause mortality (1373 vs 646 deaths per 100 000; adjusted RR, 2.00 [95% CI, 1.93-2.08]), which was higher in rural areas than in urban areas (rural RR, 2.17 [95% CI, 2.07-2.29]; urban RR, 1.83 [95% CI, 1.73-1.94]). Presence of diabetes was associated with increased mortality from ischemic heart disease (3287 deaths; RR, 2.40 [95% CI, 2.19-2.63]), stroke (4444 deaths; RR, 1.98 [95% CI, 1.81-2.17]), chronic liver disease (481 deaths; RR, 2.32 [95% CI, 1.76-3.06]), infections (425 deaths; RR, 2.29 [95% CI, 1.76-2.99]), and cancer of the liver (1325 deaths; RR, 1.54 [95% CI, 1.28-1.86]), pancreas (357 deaths; RR, 1.84 [95% CI, 1.35-2.51]), female breast (217 deaths; RR, 1.84 [95% CI, 1.24-2.74]), and female reproductive system (210 deaths; RR, 1.81 [95% CI, 1.20-2.74]). For chronic kidney disease (365 deaths), the RR was higher in rural areas (18.69 [95% CI, 14.22-24.57]) than in urban areas (6.83 [95% CI, 4.73-9.88]). Among those with diabetes, 10% of all deaths (16% rural; 4% urban) were due to definite or probable diabetic ketoacidosis or coma (408 deaths). Among adults in China, diabetes was associated with increased mortality from a range of cardiovascular and noncardiovascular diseases. Although diabetes was more common in urban areas, it was associated with greater excess mortality in rural areas.


Outcomes of Technical Variant Liver Transplantation versus Whole Liver Transplantation for Pediatric Patients: A Meta-Analysis.
To overcome the shortage of appropriate-sized whole liver grafts for children, technical variant liver transplantation has been practiced for decades. We perform a meta-analysis to compare the survival rates and incidence of surgical complications between pediatric whole liver transplantation and technical variant liver transplantation. To identify relevant studies up to January 2014, we searched PubMed/Medline, Embase, and Cochrane library databases. The primary outcomes measured were patient and graft survival rates, and the secondary outcomes were the incidence of surgical complications. The outcomes were pooled using a fixed-effects model or random-effects model. The one-year, three-year, five-year patient survival rates and one-year, three-year graft survival rates were significantly higher in whole liver transplantation than technical variant liver transplantation (OR = 1.62, 1.90, 1.65, 1.78, and 1.62, respectively, p<0.05). There was no significant difference in five-year graft survival rate between the two groups (OR = 1.47, p = 0.10). The incidence of portal vein thrombosis and biliary complications were significantly lower in the whole liver transplantation group (OR = 0.45 and 0.42, both p<0.05). The incidence of hepatic artery thrombosis was comparable between the two groups (OR = 1.21, p = 0.61). Pediatric whole liver transplantation is associated with better outcomes than technical variant liver transplantation. Continuing efforts should be made to minimize surgical complications to improve the outcomes of technical variant liver transplantation.


PPARβ/δ governs Wnt signaling and bone turnover.
Peroxisome proliferator-activated receptors (PPARs) act as metabolic sensors and central regulators of fat and glucose homeostasis. Furthermore, PPARγ has been implicated as major catabolic regulator of bone mass in mice and humans. However, a potential involvement of other PPAR subtypes in the regulation of bone homeostasis has remained elusive. Here we report a previously unrecognized role of PPARβ/δ as a key regulator of bone turnover and the crosstalk between osteoblasts and osteoclasts. In contrast to activation of PPARγ, activation of PPARβ/δ amplified Wnt-dependent and β-catenin-dependent signaling and gene expression in osteoblasts, resulting in increased expression of osteoprotegerin (OPG) and attenuation of osteoblast-mediated osteoclastogenesis. Accordingly, PPARβ/δ-deficient mice had lower Wnt signaling activity, lower serum concentrations of OPG, higher numbers of osteoclasts and osteopenia. Pharmacological activation of PPARβ/δ in a mouse model of postmenopausal osteoporosis led to normalization of the altered ratio of tumor necrosis factor superfamily, member 11 (RANKL, also called TNFSF11) to OPG, a rebalancing of bone turnover and the restoration of normal bone density. Our findings identify PPARβ/δ as a promising target for an alternative approach in the treatment of osteoporosis and related diseases.


Evaluation of Ostrinia nubilalis (Lepidoptera: Crambidae) neonate preferences for corn and weeds in corn.
Choice tests were conducted to determine feeding preferences of European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), neonates for 15 species of plants. Percentage of neonates accepting (found on) each leaf disc after 24 h was measured using choice tests. Initially, nine species of plants were evaluated. The following year, 10 plant species were evaluated during O. nubilalis first generation and 11 species during the second generation. Pennsylvania smartweed, Polygonum pennsylvanicum (L.), had the highest percentage of neonates accepting leaf discs in both years. Other plants with high acceptance rates included swamp smartweed, Polygonum amphibium L.; velvetleaf, Abutilon theophrasti Medicus; cocklebur, Xanthium strumarium L.; and yellow foxtail, Setaria glauca (L.). Corn, Zea mays L., consistently had low percentages of neonates accepting leaf discs along with common waterhemp, Amaranthus rudis Sauer. Implications these results may have on O. nubilalis host plant selection in central Iowa's corn dominated landscape are considered.


Avoiding cytotoxicity of transposases by dose-controlled mRNA delivery.
The Sleeping Beauty (SB) transposase and its newly developed hyperactive variant, SB100X, are of increasing interest for genome modification in experimental models and gene therapy. The potential cytotoxicity of transposases requires careful assessment, considering that residual integration events of transposase expression vectors delivered by physicochemical transfection or episomal retroviral vectors may lead to permanent transposase expression and resulting uncontrollable transposition. Comparing retrovirus-based approaches for delivery of mRNA, episomal DNA or integrating DNA, we found that conventional SB transposase, SB100X and a newly developed codon-optimized SB100Xo may trigger premitotic arrest and apoptosis. Cell stress induced by continued SB overexpression was self-limiting due to the induction of cell death, which occurred even in the absence of a co-transfected transposable element. The cytotoxic effects of SB transposase were strictly dose dependent and heralded by induction of p53 and c-Jun. Inactivating mutations in SB's catalytic domain could not abrogate cytotoxicity, suggesting a mechanism independent of DNA cleavage activity. An improved approach of retrovirus particle-mediated mRNA transfer allowed transient and dose-controlled expression of SB100X, supported efficient transposition and prevented cytotoxicity. Transposase-mediated gene transfer can thus be tuned to maintain high efficiency in the absence of overt cell damage.


A study of microemulsions as prolonged-release injectables through in-situ phase transition.
Microemulsions (MEs) have been studied extensively as colloidal carriers for the delivery of both water-soluble and lipid-soluble drugs. Our previous study showed that addition of water to ME formulations resulted in phase transition to either liquid crystal (LC) or coarse emulsion (CE). The aim of this study was to investigate whether these MEs could be used as drug delivery vehicles for prolonged release through in-situ phase transition following extravascular injection. Three ME formulations from the same pseudo-ternary phase diagram were investigated with respect to their phase transition behavior, and in-vivo drug release; a coarse emulsion-forming ME (CE-ME), an oil rich LC-forming ME (LC-ME1), and an oil poor LC-forming ME (LC-ME2). CE-ME was a W/O ME and both LC-MEs were O/W type. The release profiles of (99m)Tc labeled MEs following subcutaneous (SC) injection in rabbits were investigated with gamma-scintigraphy. The CE-ME dispersed readily in water, forming a CE, whereas the LC-forming MEs formed 'depots' in water. Polarized microscopy revealed a LC boundary spontaneously formed at the water/ME interface for the LC-MEs with the LC-ME2 forming a substantially thicker LC layer. The CE resulting from the water-induced transition of the CE-forming ME had a higher viscosity than the MEs, but lower than the LCs resulted from LC-MEs. Compared to LC-ME1, LC-ME2 underwent more rapid phase transition and the resultant LC had significant higher viscosity. The LCs formed from both ME formulations exhibited pseudoplastic properties; increasing the shear rate decreased the apparent viscosity exponentially. Following SC injection into the animal thigh, the LC-MEs had more prolonged release of (99m)Tc in a first-order manner, than CE-ME. The oil poor LC-ME2 had the slowest release with a t1/2 of 77min, 2.3 times longer than the oil rich LC-ME1; consistent with the thickness of LC layer formation observed in-vitro and their relative viscosities. In conclusion, the present in-vivo study has demonstrated the application of MEs as extravascular injectable drug delivery systems for sustained release. The retention of the vehicles at the injection site and the release rate were determined predominantly by their phase transition rather than ME type or oil content.


Regulation of intramembranous absorption and amniotic fluid volume by constituents in fetal sheep urine.
Our objective was to test the hypothesis that fetal urine contains a substance(s) that regulates amniotic fluid volume by altering the rate of intramembranous absorption of amniotic fluid. In late gestation ovine fetuses, amniotic fluid volumes, urine, and lung liquid production rates, swallowed volumes and intramembranous volume and solute absorption rates were measured over 2-day periods under control conditions and when urine was removed and continuously replaced at an equal rate with exogenous fluid. Intramembranous volume absorption rate decreased by 40% when urine was replaced with lactated Ringer solution or lactated Ringer solution diluted 50% with water. Amniotic fluid volume doubled under both conditions. Analysis of the intramembranous sodium and chloride fluxes suggests that the active but not passive component of intramembranous volume absorption was altered by urine replacement, whereas both active and passive components of solute fluxes were altered. We conclude that fetal urine contains an unidentified substance(s) that stimulates active intramembranous transport of amniotic fluid across the amnion into the underlying fetal vasculature and thereby functions as a regulator of amniotic fluid volume.