Tomosynthesis imaging: at a translational crossroads.
Tomosynthesis is a decades-old technique for section imaging that has seen a recent upsurge in interest due to its promise to provide three-dimensional information at lower dose and potentially lower cost than CT in certain clinical imaging situations. This renewed interest in tomosynthesis began in the late 1990s as a new generation of flat-panel detectors became available; these detectors were the one missing piece of the picture that had kept tomosynthesis from enjoying significant utilization earlier. In the past decade, tomosynthesis imaging has been investigated in a variety of clinical imaging situations, but the two most prominent have been in breast and chest imaging. Tomosynthesis has the potential to substantially change the way in which breast cancer and pulmonary nodules are detected and managed. Commercial tomosynthesis devices are now available or on the horizon. Many of the remaining research activities with tomosynthesis will be translational in nature and will involve physicist and clinician alike. This overview article provides a forward-looking assessment of the translational questions facing tomosynthesis imaging and anticipates some of the likely research and clinical activities in the next five years.


Flying high--assessing the use of the aerosphere by bats.
Bats feature prominently among organisms that occupy the aerosphere as they extensively use this environment for foraging, but also for dispersal, migration, and behavioral interactions. Differential use of the aerosphere is an important factor structuring bat assemblages, with species exhibiting distinct morphological, physiological, and sensory adaptations to different habitat types. This necessitates comprehensive sampling methodologies such as combined ground-level and canopy-level mist netting as well as acoustic monitoring to assess the presence, diversity, and activity of different functional groups of species adequately. Recent technological advances in acoustic detection and in methods of analysis, coupled with the expansion of libraries of echolocation calls for species identification, now allow for the reliable quantification of species numbers and activity of the scarcely known group of aerial insectivorous bats, particularly in species-rich tropical assemblages. We provide a brief, exemplary overview of recent studies on bats conducted in Panamá to demonstrate the necessity of comprehensive sampling methods and application of new technologies in order to adequately depict assemblage composition and responses of bats to structural changes in habitats induced by fragmentation. In addition to acoustic methods, miniaturization of radio transmitters has provided new insights into the patterns of spatial use of the aerosphere by bats and has identified species-specific differences in mobility as one of the important traits that determines bats' reactions to anthropogenic alterations of the landscape. Following the goals of the symposium on aeroecology, we propose new avenues of research for probing the aerosphere. We discuss how integration of a diverse array of remote sensing tools with data on species distribution and species traits, such as mobility and edge-sensitivity, might provide novel opportunities for the development, and application of conservation-oriented monitoring systems.


Adaptive soft tissue deformation for a virtual reality surgical trainer.
Real time tissue deformation is an important aspect of interactive virtual reality (VR) environments such as medical trainers. Most approaches in deformable modelling use a fixed space discretization. A surgical trainer requires high plausibility of the deformations especially in the area close to the instrument. As the area of intervention is not known a priori, adaptive techniques have to be applied. We present an approach for real time deformation of soft tissue based on a regular FEM mesh of cube elements as opposed to a mesh of tetrahedral elements used by the majority of soft tissue simulators. A regular mesh structure simplifies the local refinement operation as the elements topology and stiffness are known implicitly. We propose an octree-based adaptive multiresolution extension of our basic approach. The volumetric representation of the deformed object is created automatically from medical images or by voxelization of a surface model. The resolution of the volumetric representation is independent of the surface geometry resolution. The surface is deformed according to the simulation performed on the underlying volumetric mesh.


Higher chylomicron remnants and LDL particle numbers associate with CD36 SNPs and DNA methylation sites that reduce CD36.
Cluster of differentiation 36 (CD36) variants influence fasting lipids and risk of metabolic syndrome, but their impact on postprandial lipids, an independent risk factor for cardiovascular disease, is unclear. We determined the effects of SNPs within a ∼410 kb region encompassing CD36 and its proximal and distal promoters on chylomicron (CM) remnants and LDL particles at fasting and at 3.5 and 6 h following a high-fat meal (Genetics of Lipid Lowering Drugs and Diet Network study, n = 1,117). Five promoter variants associated with CMs, four with delayed TG clearance and five with LDL particle number. To assess mechanisms underlying the associations, we queried expression quantitative trait loci, DNA methylation, and ChIP-seq datasets for adipose and heart tissues that function in postprandial lipid clearance. Several SNPs that associated with higher serum lipids correlated with lower adipose and heart CD36 mRNA and aligned to active motifs for PPARγ, a major CD36 regulator. The SNPs also associated with DNA methylation sites that related to reduced CD36 mRNA and higher serum lipids, but mixed-model analyses indicated that the SNPs and methylation independently influence CD36 mRNA. The findings support contributions of CD36 SNPs that reduce adipose and heart CD36 RNA expression to inter-individual variability of postprandial lipid metabolism and document changes in CD36 DNA methylation that influence both CD36 expression and lipids.


Alterations of ubiquitin related proteins in the pathology and development of schizophrenia: Evidence from human and animal studies.
Gene expression analyses in post-mortem schizophrenia brains suggest that a number of ubiquitin proteasome system (UPS) genes are associated with schizophrenia; however the status of UPS proteins in the schizophrenia brain is largely unknown. Ubiquitin related proteins are inherently involved in memory, neuronal survival and morphology, which are processes implicated in neurodevelopmental disorders such as schizophrenia. We examined levels of five UPS proteins (Protein Inhibitor of Activated STAT2 [PIAS2], F-Box and Leucine rich repeat protein 21 [FBXL21], Mouse Double Minute 2 homolog [MDM2], Ubiquitin Carboxyl-Terminal Hydrolase-L1 [UCHL1] and Ubiquitin Conjugating Enzyme E2D1 [UBE2D1]) involved in these neuronal processes, within the dorsolateral prefrontal cortex of post-mortem schizophrenia subjects and matched controls (n = 30/group), in addition to across neurodevelopmental time-points (juvenile, adolescent and adult stages of life), utilizing a well-established neurodevelopmental phencyclidine (PCP) animal model of schizophrenia. We observed significant reductions in PIAS2, FBXL21 and MDM2 in schizophrenia subjects compared to controls (p-values ranging from 0.002 to 0.004). In our developmental PCP model, MDM2 protein was significantly reduced in adult PCP-treated rats compared to controls (p = 0.034). Additionally, FBXL21 (p = 0.022) and UCHL1 (p = 0.022) were significantly decreased, whilst UBE2D1 was increased (p = 0.022), in juvenile phencyclidine-treated rats compared to controls. This is the first study reporting alterations of UPS proteins in post-mortem human schizophrenia subjects and in a neurodevelopmental model of schizophrenia. The findings from this study provide strong support for a role of these UPS proteins in the pathology and development of schizophrenia.


Srp20 regulates TrkB pre-mRNA splicing to generate TrkB-Shc transcripts with implications for Alzheimer's disease.
Previously, we reported elevated levels of the neuron-specific tropomyosin receptor kinase B (TrkB) transcript, TrkB- sarc homology containing (Shc) in the hippocampus of Alzheimer's disease (AD) brains. In this study, we determined how TrkB-Shc transcripts are increased in AD. Utilizing a TrkB minigene transiently transfected into SHSY5Y cells, we found increased exon 19 inclusion in TrkB minigene transcripts (to generate TrkB-Shc) following cellular exposure to amyloid beta 1-42 (Αβ(42)). As this suggested altered TrkB pre-mRNA splicing in AD, we conducted an in silico screening for putative splice regulatory protein-binding sites in the intron/exon splice regulatory regions of exons 18 and 19 of the TrkB gene and then assessed their gene expression profiles using a microarray database of control/AD post-mortem human hippocampal brain tissue. We found significant changes in serine/arginine protein 20 (Srp20) gene expression in AD cases and confirmed this using a second cohort of control/AD. In vitro, we found increased Srp20 mRNA levels in SHSY5Y cells treated with Αβ(42) fibrils. Moreover, Srp20 over-expression was found to increase exon 19 inclusion in TrkB minigene transcripts and ratio of endogenous TrkB-Shc:TrkB-TK+ mRNA expression. Conversely, Srp20 expression knockdown produced the opposite effects. Our findings suggest that dysregulation of factors regulating TrkB pre-mRNA splicing may contribute to gene expression changes that occur in AD.


Enhanced immunogenicity of CTL antigens through mutation of the CD8 binding MHC class I invariant region.
CD8(+) cytotoxic T lymphocytes (CTL) are key determinants of immunity to intracellular pathogens and neoplastic cells. Recognition of specific antigens in the form of peptide-MHC class I complexes (pMHCI) presented on the target cell surface is mediated by T cell receptor (TCR) engagement. The CD8 coreceptor binds to invariant domains of pMHCI and facilitates antigen recognition. Here, we investigate the biological effects of a Q115E substitution in the alpha2 domain of human leukocyte antigen (HLA)-A*0201 that enhances CD8 binding by approximately 50% without altering TCR/pMHCI interactions. Soluble and cell surface-expressed forms of Q115E HLA-A*0201 exhibit enhanced recognition by CTL without loss of specificity. These CD8-enhanced antigens induce greater CD3 zeta chain phosphorylation in cognate CTL leading to substantial increases in cytokine production, proliferation and priming of naive T cells. This effect provides a fundamental new mechanism with which to enhance cellular immunity to specific T cell antigens.


Monte Carlo simulation of the atmospheric point-spread function with an application to correction for the adjacency effect.
Monte Carlo techniques are used to simulate atmospheric point-spread functions (PSF's) that are appropriate for the viewing geometries typical of the Airborne Visible-Infrared Imaging Spectrometer (AVIRIS). A model sensor is located at an altitude of 20 km and views a Lambertian surface through a horizontally homogeneous and vertically stratified atmosphere. Simulations show the effects on the PSF of variation of the aerosol phase function, the aerosol optical thickness, the sensor viewing angle, and the wavelength. An algorithm that uses the PSF to correct high-contrast images for adjacency effects is developed and applied to an AVIRIS image of Big Pine Key in the Florida Keys. A method to approximate the atmospheric PSF's without the need to resort to a Monte Carlo simulation is described. Correction of the AVIRIS image through the use of the approximated PSF is consistent with a previous correction. Error analysis is difficult and scene dependent; however, the correction algorithm is shown to be capable of indicating regions of high-contrast images in which conventional estimates of surface-leaving radiance are likely to be unreliable due to adjacency effects.


Impact of the 2002 Canadian forest fires on particulate matter air quality in Baltimore city.
With increasing evidence of adverse health effects associated with particulate matter (PM), the exposure impact of natural sources, such as forest fires, has substantial public health relevance. In addition to the threat to nearby communities, pollutants released from forest fires can travel thousands of kilometers to heavily populated urban areas. There was a dramatic increase in forest fire activity in the province of Quebec, Canada, during July 2002. The transport of PM released from these forest fires was examined using a combination of a moderate-resolution imaging spectroradiometer satellite image, back-trajectories using a hybrid single-particle Lagrangian integrated trajectory, and local light detection and ranging measurements. Time- and size-resolved PM was evaluated at three ambient and four indoor measurement sites using a combination of direct reading instruments (laser, time-of-flight aerosol spectrometer, nephelometer, and an oscillating microbalance). The transport and monitoring results consistently identified a forest fire related PM episode in Baltimore that occurred the first weekend of July 2002 and resulted in as much as a 30-fold increase in ambientfine PM. On the basis of tapered element oscillating microbalance measurements, the 24 h PM25 concentration reached 86 microg/m3 on July 7, 2002, exceeding the 24 h national ambient air quality standard. The episode was primarily comprised of particles less than 2.5 microm in aerodynamic diameter, highlighting the preferential transport of the fraction of PM that is of greatest health concern. Penetration of the ambient episode indoors was efficient (median indoor-to-outdoor ratio 0.91) such that the high ambient levels were similarly experienced indoors. These results are significant in demonstrating the impact of a natural source thousands of kilometers away on ambient levels of and potential exposures to air pollution within an urban center. This research highlights the significance of transboundary air pollution and the need for studies that assess the public health impacts associated with such sources and transport processes.


Mfsd2a Attenuates Blood-Brain Barrier Disruption After Sub-arachnoid Hemorrhage by Inhibiting Caveolae-Mediated Transcellular Transport in Rats.
Blood-brain barrier (BBB) disruption is one of the critical mechanisms of brain injury induced by subarachnoid hemorrhage (SAH). Past studies have often focused on the tight junctions of endothelial cells. However, low transcellular transport levels also play an important role in the normal functioning of the BBB. Major facilitator superfamily domain-containing 2a (Mfsd2a) has been demonstrated to be essential for the maintenance of the normal BBB. Our present study aimed to explore the roles and mechanisms of Mfsd2a in BBB disruption after SAH. In this study, a prechiasmatic cistern single-injection model was used to produce experimental SAH in Sprague-Dawley rats. Specific small-interfering RNA and plasmids were used to downregulate and upregulate the expression of Mfsd2a prior to assessments in our SAH model. Omega-3 fatty acid deficiency diet was used to reduce DHA in rat brain. The expression level of Mfsd2a decreased significantly after SAH and reached its lowest level at 72 h post-SAH, which then gradually recovered. At 72 h after SAH, BBB function was disrupted; upregulation of Mfsd2a reversed this damage, whereas downregulation of Mfsd2a exacerbated this damage. These effects were primarily mediated through transcellular transport, especially for changes in caveolae compared to those of tight junctions. After stopping the supply of omega-3 fatty acids, the effect of Mfsd2a on inhibition of caveolae and protection of the blood-brain barrier was eliminated. Taken together, Mfsd2a inhibits caveolae-based transcellular transport by transporting omega-3 fatty acids to protect the BBB after SAH.


Autotrophic nitrogen removal after ureolytic phosphate precipitation to remove both endogenous and exogenous nitrogen.
Anaerobic digestion yields effluents rich in ammonium and phosphate and poor in biodegradable organic carbon, thereby making them less suitable for conventional biological nitrogen and phosphorus removal. In addition, the demand for fertilizers is increasing, energy prices are rising and global phosphate reserves are declining. This requires both changes in wastewater treatment technologies and implementation of new processes. In this contribution a description is given of the combination of a ureolytic phosphate precipitation (UPP) and an autotrophic nitrogen removal (ANR) process on the anaerobic effluent of a potato processing company. The results obtained show that it is possible to recover phosphate as struvite and to remove the nitrogen with the ANR process. The ANR process was performed in either one or two reactors (partial nitritation + Anammox). The one-reactor configuration operated stably when the dissolved oxygen was kept between 0.1 and 0.35 mg L(-1). The best results for the two-reactor system were obtained when part of the effluent of the UPP was fully nitrified in a nitritation reactor and mixed in a 3:5 volumetric ratio with untreated ammonium-containing effluent. A phosphate and nitrogen removal efficiency of respectively 83 ± 1% and of 86 ± 7% was observed during this experiment.


Patterns of Ancestral Animal Codon Usage Bias Revealed through Holozoan Protists.
Choanoflagellates and filastereans are the closest known single celled relatives of Metazoa within Holozoa and provide insight into how animals evolved from their unicellular ancestors. Codon usage bias has been extensively studied in metazoans, with both natural selection and mutation pressure playing important roles in different species. The disparate nature of metazoan codon usage patterns prevents the reconstruction of ancestral traits. However, traits conserved across holozoan protists highlight characteristics in the unicellular ancestors of Metazoa. Presented here are the patterns of codon usage in the choanoflagellates Monosiga brevicollis and Salpingoeca rosetta, as well as the filasterean Capsaspora owczarzaki. Codon usage is shown to be remarkably conserved. Highly biased genes preferentially use GC-ending codons, however there is limited evidence this is driven by local mutation pressure. The analyses presented provide strong evidence that natural selection, for both translational accuracy and efficiency, dominates codon usage bias in holozoan protists. In particular, the signature of selection for translational accuracy can be detected even in the most weakly biased genes. Biased codon usage is shown to have coevolved with the tRNA species, with optimal codons showing complementary binding to the highest copy number tRNA genes. Furthermore, tRNA modification is shown to be a common feature for amino acids with higher levels of degeneracy and highly biased genes show a strong preference for using modified tRNAs in translation. The translationally optimal codons defined here will be of benefit to future transgenics work in holozoan protists, as their use should maximise protein yields from edited transgenes.


Prognosis of bulky Hodgkin's disease treated with chemotherapy alone or combined with radiotherapy.
The medical records of 396 consecutive patients with a histological diagnosis of Hodgkin's disease were reviewed to assess the prognostic importance of bulky and non-bulky lymphomas. The presence of large lymphadenopathy failed to affect significantly the seven-year results in terms of complete remission (bulky 81.1% v. non-bulky 86.2%), freedom from progression (60.7% v. 65.6%), relapse-free survival (75.1% v. 76.5%) and overall survival (62.7% v. 68.9%). It is noteworthy that in all subsets, ABVD (Adriamycin + bleomycin + vinblastine + dacarbazine), either combined with irradiation or alternated with MOPP (mechlorethamine + vincristine + procarbazine + prednisone), yielded superior results compared with MOPP with or without irradiation. Given the prognostic importance of various bulky sites, the presence of large lymphadenopathy in anatomic regions other than the mediastinum failed to affect results adversely. On the contrary, in patients in stages IIB-IIIA-IIIB, treated with combined modality, the presence of bulky mediastinal involvement did influence prognosis compared with patients with positive but non-bulky mediastinum. At seven years the results were 60.2% v. 79.9% for freedom from progression, 73.2% v. 89.9% for relapse-free survival and 64.8% v. 87.1% for total survival, respectively (P less than 0.03). By contrast, in patients with stage IV disease the extent of mediastinal involvement did not affect results. Nonetheless, the frequency of intrathoracic relapses was higher (26.7%) in patients given chemotherapy alone compared with patients treated with combined modality (11.5%).


Liver transplantation from living donors with Gilbert's syndrome is a safe procedure for both donors and recipients.
Liver transplantation (LT) has become a favorable therapeutic option for patients with end-stage liver diseases. Gilbert's syndrome (GS) is a benign condition characterized by intermittent mild jaundice due to unconjugated hyperbilirubinemia. It is not obvious whether living-donor liver transplantation (LDLT) from a donor with GS could result in a normal outcome for both the recipient and the donor. We aimed to determine whether right lobe hepatectomy is a safe procedure for living donors with GS and LT recipients. Between September 2011 and March 2015, 305 LDLT procedures using right lobe grafts were performed at Atasehir Memorial Hospital, Istanbul, Turkey. Nineteen of 305 LT candidates who had been diagnosed with GS were included in the current study. After a 12-h overnight fast, total and indirect bilirubin levels of donors and recipients were measured. The median follow-up after transplant was 16 months (range 3-36 months). The median age of donors was 25 (range 20-55 yr). Four donors (21%) were female, and 15 donors (89%) were male. The median age of donors was 51 (range 23-68 yr). Eleven recipients (57%) were female, and 8 (43%) were male. The median preoperative total bilirubin level of donors was 1.69 mg/dL (range 1.26-2.43 mg/dL) (normal range <1.2 mg/dL). The median total bilirubin level of donors on postoperative day 7 was 1.04 mg/dL (range 0.71-3.23 mg/dL). As our study has included a large number of donors with GS, it produced reliable evidence that right lobe hepatectomy is a safe procedure for living donors with GS and LT recipients.


40Ar/39Ar dates from the West Siberian Basin: Siberian flood basalt province doubled.
Widespread basaltic volcanism occurred in the region of the West Siberian Basin in central Russia during Permo-Triassic times. New 40Ar/39Ar age determinations on plagioclase grains from deep boreholes in the basin reveal that the basalts were erupted 249.4 +/- 0.5 million years ago. This is synchronous with the bulk of the Siberian Traps, erupted further east on the Siberian Platform. The age and geochemical data confirm that the West Siberian Basin basalts are part of the Siberian Traps and at least double the confirmed area of the volcanic province as a whole. The larger area of volcanism strengthens the link between the volcanism and the end-Permian mass extinction.


Standardized evaluation framework for evaluating coronary artery stenosis detection, stenosis quantification and lumen segmentation algorithms in computed tomography angiography.
Though conventional coronary angiography (CCA) has been the standard of reference for diagnosing coronary artery disease in the past decades, computed tomography angiography (CTA) has rapidly emerged, and is nowadays widely used in clinical practice. Here, we introduce a standardized evaluation framework to reliably evaluate and compare the performance of the algorithms devised to detect and quantify the coronary artery stenoses, and to segment the coronary artery lumen in CTA data. The objective of this evaluation framework is to demonstrate the feasibility of dedicated algorithms to: (1) (semi-)automatically detect and quantify stenosis on CTA, in comparison with quantitative coronary angiography (QCA) and CTA consensus reading, and (2) (semi-)automatically segment the coronary lumen on CTA, in comparison with expert's manual annotation. A database consisting of 48 multicenter multivendor cardiac CTA datasets with corresponding reference standards are described and made available. The algorithms from 11 research groups were quantitatively evaluated and compared. The results show that (1) some of the current stenosis detection/quantification algorithms may be used for triage or as a second-reader in clinical practice, and that (2) automatic lumen segmentation is possible with a precision similar to that obtained by experts. The framework is open for new submissions through the website, at http://coronary.bigr.nl/stenoses/.


Retrievable IVC Filters: Comprehensive Review of Device-related Complications and Advanced Retrieval Techniques.
Use of retrievable inferior vena cava filters (RIVCFs) has grown exponentially since their introduction into clinical practice, but many of these devices are not retrieved. Some are not retrieved due to poor clinical follow-up, but other devices remain in situ for extended periods because they present significant technical challenges during retrieval. Because of these and other factors, many of these devices were thus left in place permanently. However, recent data have placed a renewed emphasis on device retrieval due to increased risk of RIVCF-related complications, which are positively correlated with filter dwell time. Development of advanced filter retrieval techniques has had a significant impact on the removal of embedded RIVCFs, permitting retrieval of the majority of devices. The purpose of this article is to present an imaging and data review of the dominant device-related factors that complicate RIVCF retrieval and to describe the relevant advanced retrieval techniques to manage these factors. RIVCF imaging is frequently encountered in daily clinical practice via various imaging modalities. Therefore, diagnostic radiologists can play a vital role in identifying filter-related issues. Familiarity with the context for managing these issues in the interventional suite is essential for improving triage and care of patients with RIVCFs. © RSNA, 2017.


Novel Coating of Surgical Suture Confers Antimicrobial Activity Against Porphyromonas gingivalis and Enterococcus faecalis.
The oral cavity is colonized by >10(9) bacteria, many of which can increase heart disease risk when seeded into the bloodstream. Most dentoalveolar surgeries require the use of surgical sutures. Suture placement and removal can increase the risk of postoperative infection and bacteremia. The aim of this study is to evaluate the antimicrobial activity of a novel quaternary ammonium compound, K21, when coated on different suture materials. The periodontal pathogen Porphyromonas gingivalis and the endodontic species Enterococcus faecalis were grown to early log phase and inoculated on enriched Brucella blood agar, on which were placed identical lengths of surgical suture (chromic gut, polyester suture, silk, and nylon suture) and control unwaxed dental floss impregnated with K21 at 5%, 10%, 20%, and 25% volume/volume in ethanol vehicle. Controls included the following: 1) sutures treated with vehicle; 2) untreated sutures; and 3) unwaxed floss. Zones of inhibition in millimeters were measured at five randomized sites per suture/floss for each concentration and material used. Mean ± SD of zones of inhibition were calculated, and analysis of variance (P <0.05) was used to determine whether differences were statistically significant. The results indicate that K21-coated suture at concentrations ranging from 5% to 25%, depending on the type of suture, have antimicrobial activity for P. gingivalis and E. faecalis. Nylon suture coated with K21 at 5%, 10%, 20%, and 25% resulted in zones ranging from 3 to 11 mm. Polyester suture was more effective at lower K21 concentrations with 5% (P = 0.0031), 10% (P = 0.0011), and 20% (P = 0.0002), yielding 7.5, 8.3, and 10.5 mm zones of inhibition. K21-coated silk suture yielded significant zones of inhibition at 25% (P <0.0001), whereas chromic gut was effective at K21 concentrations of 5% (P = 0.0081) and 25% (P <0.0001). It is concluded that K21-coated surgical sutures have antimicrobial activity for bacterial species of direct relevance to postoperative infection and bacteremia.


Meditation and Cardiovascular Health in the US.
The 2017 American Heart Association Scientific Statement on meditation and cardiovascular risk suggested that meditation may be considered as an adjunct to guideline-directed cardiovascular risk-reduction interventions. Meditation could potentially increase physical and mental relaxation, leading to improved outcomes after a major cardiovascular event. We hypothesized that meditation is associated with lower cardiovascular risk in the US general population. Using data from the 2012 and 2017 National Health Interview Survey, we identified all patients with hypercholesterolemia, systemic hypertension (SH), diabetes mellitus (DM), stroke, and coronary artery disease (CAD), as well as those who reported that they meditate. Multivariable logistic regression analyses were performed to evaluate the association between meditation and risk of hypercholesterolemia, SH, DM, stroke, and CAD, adjusting for potential confounders. Of 61,267 the National Health Interview Survey participants, 5,851 (9.6%) participated in some form of meditation. After adjusting for age, gender, body mass index, race, marital status, cigarette smoking, sleeping duration, and depression, meditation was independently associated with a lower prevalence of hypercholesterolemia (odds ratio [OR] 0.65; 95% confidence interval [CI] 0.54 to 0.79; p = 0.001), SH (OR 0.86; 95% CI 0.75 to 0.99; p = 0.04), diabetes (OR 0.70; 95% CI 0.59 to 0.84; p = 0.0001), stroke (OR 0.76; 95% CI 0.58 to 0.99; p = 0.04), or CAD (OR 0.51; 95% CI 0.39 to 0.66; p <0.001), compared with those who did not meditate. In conclusion, using a large national database, we found that meditation is associated with a lower prevalence of cardiovascular risks factors and disease.


Validity and reliability of a smartphone application for the assessment of penile deformity in Peyronie's disease.
Available methods to evaluate men with Peyronie's disease (PD) are limited by the inability to accurately and reproducibly measure penile deformity. AIM.: The study aims to evaluate the performance of a smartphone application for the measurement of penile curvature and narrowing. A smartphone application, the University of Washington Peyronie's Examination Network (UWPEN), was developed for this purpose. To assess penile curvature, 15 single cylinders of malleable penile prostheses were molded to varying curvature angles. Three blinded observers nonsequentially measured the angle of curvature for each prosthetic cylinder using a protractor, goniometer, and UWPEN. To assess girth narrowing, six clay models of the penile shaft were constructed to represent conditions of normal, partial hourglass, circumferential hourglass, and pencil narrowing. Girth was measured using a ruler and UWPEN by the same blinded observers. Statistical analyses compared intertest, interobserver, and intraobserver reliability using the interclass correlation coefficient (ICC). An ICC above 0.75 indicates excellent reproducibility among measurements. Intertest reliability for angle measurements yielded an ICC for the three methods of 1.000. Separately, the ICC for UWPEN vs. the goniometer and protractor was 0.999 and 0.999, respectively. The interobserver ICC for UWPEN, goniometer, and protractor was 0.998, 0.999, and 1.000, respectively. Intertest reliability for girth narrowing measurements yielded an ICC of 0.991. The interobserver ICC for girth narrowing for UWPEN and the ruler was 0.978 and 0.986, respectively. Intraobserver ICC for angle measurements and girth narrowing showed high reliability for all observers and methods. The performance of UWPEN is comparable with and highly correlated with angle measurements obtained from the goniometer and protractor as well as with girth narrowing measurements obtained from a ruler. Measurements are reproducible among different observers. UWPEN may provide a noninvasive, accurate, reliable, and widely accessible method to characterize and track PD over time.


Novel 8-nitroquinolin-2(1H)-ones as NTR-bioactivated antikinetoplastid molecules: Synthesis, electrochemical and SAR study.
To study the antiparasitic 8-nitroquinolin-2(1H)-one pharmacophore, a series of 31 derivatives was synthesized in 1-5 steps and evaluated in vitro against both Leishmania infantum and Trypanosoma brucei brucei. In parallel, the reduction potential of all molecules was measured by cyclic voltammetry. Structure-activity relationships first indicated that antileishmanial activity depends on an intramolecular hydrogen bond (described by X-ray diffraction) between the lactam function and the nitro group, which is responsible for an important shift of the redox potential (+0.3 V in comparison with 8-nitroquinoline). With the assistance of computational chemistry, a set of derivatives presenting a large range of redox potentials (from -1.1 to -0.45 V) was designed and provided a list of suitable molecules to be synthesized and tested. This approach highlighted that, in this series, only substrates with a redox potential above -0.6 V display activity toward L. infantum. Nevertheless, such relation between redox potentials and in vitro antiparasitic activities was not observed in T. b. brucei. Compound 22 is a new hit compound in the series, displaying both antileishmanial and antitrypanosomal activity along with a low cytotoxicity on the human HepG2 cell line. Compound 22 is selectively bioactivated by the type 1 nitroreductases (NTR1) of L. donovani and T. brucei brucei. Moreover, despite being mutagenic in the Ames test, as most of nitroaromatic derivatives, compound 22 was not genotoxic in the comet assay. Preliminary in vitro pharmacokinetic parameters were finally determined and pointed out a good in vitro microsomal stability (half-life > 40 min) and a 92% binding to human albumin.


Comparative analysis of the drought-responsive transcriptome in soybean lines contrasting for canopy wilting.
Drought stress causes significant yield losses in major oil seed crops, such as soybean [Glycine max (L.) Merr]. Few soybean lines have been identified as canopy-wilting tolerant; however, the molecular mechanism conferring tolerance is not fully understood. To understand the biological process, a whole genome transcriptome analysis was performed for leaf tissues of two contrasting soybean lines: drought-susceptible (DS) Pana and drought-tolerant (DT) PI 567690. A pairwise comparison of the DS and DT lines under drought and control conditions detected 1914 and 670 genes with a greater than two-fold change in expression under drought conditions. Pairwise treatment comparison and gene enrichment analysis on the DT line showed the down-regulation of genes associated with protein binding, hydrolase activity, carbohydrate/lipid metabolism, xyloglucan endo-transglycosylases associated with cell-wall, apoplast, and chlorophyll a/b binding proteins. On the other hand, genes that were associated with the biotic stress response, ion binding and transport, the oxido-reductive process and electron carrier activity were up-regulated. Gene enrichment analysis detected UDP glucuronosyl transferase activity-encoding genes to be differentially expressed in PI 567690 under drought stress conditions. We found valuable SNPs variation in aquaporin genes of the DT line that are conserved in known slower canopy-wilting lines, this should facilitate marker-assisted selection in soybeans with improved drought tolerance.


Effects of local authority expenditure on childhood obesity.
Under the 2013 reforms introduced by the Health and Social Care Act (2012), public health responsibilities in England were transferred from the National Health Service to local authorities (LAs). Ring-fenced grants were introduced to support the new responsibilities. The aim of our study was to test whether the level of expenditure in 2013/14 affected the prevalence of childhood obesity in 2016/17. We used National Child Measurement Programme definitions of childhood obesity and datasets. We used LA revenue returns data to derive three measures of per capita expenditure: childhood obesity (<19); physical activity (<19) and the Children's 5-19 Public Health Programme. We ran separate negative binomial models for two age groups of children (4-5 year olds; 10-11 year olds) and conducted sensitivity analyses. With few exceptions, the level of spend in 2013/14 was not significantly associated with the level of childhood obesity in 2016/17. We identified some positive associations between spend on physical activity and the Children's Public Health Programme at baseline (2013/14) and the level of childhood obesity in children aged 4-5 in 2016/17, but the effect was not evident in children aged 10-11. In both age groups, LA levels of childhood obesity in 2016/17 were significantly and positively associated with obesity levels in 2013/14. As these four cohorts comprise entirely different pupils, this underlines the importance of local drivers of childhood obesity. Higher levels of local expenditure are unlikely to be effective in reducing childhood obesity in the short term.


The octopamine receptor Octβ2R regulates ovulation in Drosophila melanogaster.
Oviposition is induced upon mating in most insects. Ovulation is a primary step in oviposition, representing an important target to control insect pests and vectors, but limited information is available on the underlying mechanism. Here we report that the beta adrenergic-like octopamine receptor Octβ2R serves as a key signaling molecule for ovulation and recruits protein kinase A and Ca(2+)/calmodulin-sensitive kinase II as downstream effectors for this activity. We found that the octβ2r homozygous mutant females are sterile. They displayed normal courtship, copulation, sperm storage and post-mating rejection behavior but were unable to lay eggs. We have previously shown that octopamine neurons in the abdominal ganglion innervate the oviduct epithelium. Consistently, restored expression of Octβ2R in oviduct epithelial cells was sufficient to reinstate ovulation and full fecundity in the octβ2r mutant females, demonstrating that the oviduct epithelium is a major site of Octβ2R's function in oviposition. We also found that overexpression of the protein kinase A catalytic subunit or Ca(2+)/calmodulin-sensitive protein kinase II led to partial rescue of octβ2r's sterility. This suggests that Octβ2R activates cAMP as well as additional effectors including Ca(2+)/calmodulin-sensitive protein kinase II for oviposition. All three known beta adrenergic-like octopamine receptors stimulate cAMP production in vitro. Octβ1R, when ectopically expressed in the octβ2r's oviduct epithelium, fully reinstated ovulation and fecundity. Ectopically expressed Octβ3R, on the other hand, partly restored ovulation and fecundity while OAMB-K3 and OAMB-AS that increase Ca(2+) levels yielded partial rescue of ovulation but not fecundity deficit. These observations suggest that Octβ2R have distinct signaling capacities in vivo and activate multiple signaling pathways to induce egg laying. The findings reported here narrow the knowledge gap and offer insight into novel strategies for insect control.


Noise-aware image deconvolution with multidirectional filters.
In this paper we propose an approach for handling noise in deconvolution algorithm based on multidirectional filters. Most image deconvolution techniques are sensitive to the noise. Even a small amount of noise will degrade the quality of image estimation dramatically. We found that by applying a directional low-pass filter to the blurred image, we can reduce the noise level while preserving the blur information in the orthogonal direction to the filter. So we apply a series of directional filters at different orientations to the blurred image, and a guided filter based edge-preserving image deconvolution is used to estimate an accurate Radon transform of the clear image from each filtered image. Finally, we reconstruct the original image using the inverse Radon transform. We compare our deconvolution algorithm with many competitive deconvolution techniques in terms of the improvement in signal-to-noise ratio and visual quality.


Protein oxidation and aging.
A number of systems that generate oxygen free radicals catalyze the oxidative modification of proteins. Such modifications mark enzymes for degradation by cytosolic neutral alkaline proteases. Protein oxidation contributes to the pool of damaged enzymes, which increases in size during aging and in various pathological states. The age-related increase in amounts of oxidized protein may reflect the age-dependent accumulation of unrepaired DNA damage that, in a random manner, affects the concentrations or activities of numerous factors that govern the rates of protein oxidation and the degradation of oxidized protein.


Hovering between Heaven and Hell: An Observational Study Focusing on the Interactions between One Woman with Schizophrenia, Dementia, and Challenging Behaviour and her Care Providers.
This case study aims to illuminate the interactions between one woman (Alice) with schizophrenia, dementia, and challenging behaviour and her professional caregivers. We performed participant observations of these interactions and conducted informal interviews at the residential home where the woman lived. The transcripts were subjected to qualitative content analysis. The results showed that the interactions between Alice and her caregivers were experienced as hovering between heaven and hell. Alice struggled to bring order into her chaotic life world by splitting herself and others, and her caregivers struggled to protect Alice's and their own dignity by limiting her challenging behaviours. They also strived to understand their own and Alice's behaviour. Current practice in caring for people with challenging behaviour usually focuses on symptom reduction through medication and behavioural modification. Instead, we suggest moving toward an understanding of the experiences behind the challenging behaviours and designing person-centred care based on each patients' reality.


Breast cancer and dietary patterns: a systematic review.
This systematic review collates research on the topic of dietary patterns and breast cancer risks. The literature search targeted epidemiological studies published up to December 2012 and was conducted using the Medline (U.S. National Library of Medicine, Bethesda MD, USA) and Lilacs (Latin American and Caribbean Health Sciences, São Paulo, Brazil) databases. The following search terms were used: breast cancer, breast neoplasm, breast carcinoma, diet, food, eating habits, dietary patterns, factor analysis, and principal component analysis. Only studies that used factor analysis techniques and/or principal component analysis were eligible, and a total of 26 studies were included. The findings of these studies suggest the Mediterranean dietary pattern and diets composed largely of vegetables, fruit, fish, and soy are associated with a decreased risk of breast cancer. There was no evidence of an association between traditional dietary patterns and risk of breast cancer, and only one study showed a significant increase in risk associated with the Western dietary pattern. Diets that include alcoholic beverages may be associated with increased risk.


Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases.
The two primary human inflammatory bowel diseases, Crohn's disease (CD) and ulcerative colitis (UC), are idiopathic relapsing disorders characterized by chronic inflammation of the intestinal tract. Although several lines of reasoning suggest that gastrointestinal (GI) microbes influence inflammatory bowel disease (IBD) pathogenesis, the types of microbes involved have not been adequately described. Here we report the results of a culture-independent rRNA sequence analysis of GI tissue samples obtained from CD and UC patients, as well as non-IBD controls. Specimens were obtained through surgery from a variety of intestinal sites and included both pathologically normal and abnormal states. Our results provide comprehensive molecular-based analysis of the microbiota of the human small intestine. Comparison of clone libraries reveals statistically significant differences between the microbiotas of CD and UC patients and those of non-IBD controls. Significantly, our results indicate that a subset of CD and UC samples contained abnormal GI microbiotas, characterized by depletion of commensal bacteria, notably members of the phyla Firmicutes and Bacteroidetes. Patient stratification by GI microbiota provides further evidence that CD represents a spectrum of disease states and suggests that treatment of some forms of IBD may be facilitated by redress of the detected microbiological imbalances.


Anti-Amoxicillin Immunoglobulin E, Histamine-2 Receptor Antagonist Therapy and Mast Cell Activation Syndrome Are Risk Factors for Amoxicillin Anaphylaxis.
β-Lactam antibiotics (mainly amoxicillin, AX) are the drugs that most frequently induce systemic drug allergy reactions. We attempted to identify the risk factors associated with systemic reactions to AX. All patients who were referred to our department for suspected hypersensitivity reactions to AX over a 6-month period were evaluated for anti-AX immunoglobulin E (IgE) levels and skin-test positivity for β-lactams. Age, sex, concomitant diseases, therapies, total IgE, serum tryptase levels and signs and symptoms suggesting mast cell activation syndrome (MCAS) were analyzed in relation to the severity of the reaction in accordance with the Mueller classification. Sixty-seven patients were selected: 39 with mild reactions such as cutaneous or gastrointestinal symptoms (grades I and II) and 28 with severe systemic reactions (grades III and IV). Anti-AX IgE levels and total IgE were significantly higher in severe reactions than in mild ones (p < 0.00005, p = 0.0037). Treatment with histamine-2 receptor antagonists (anti-H2) was significantly related to severe reactions (p = 0.007). No significant correlations were found between the severity of the reactions and dyslipidemia or levels of angiotensin-converting enzyme and tryptase. Anti-AX IgE levels were the most significant immunological parameter distinguishing patients who presented with severe reactions to AX and those with mild reactions. Higher values of total IgE, the use of gastroprotective drugs and signs and symptoms suggesting an MCAS significantly increased the odds ratio of having a severe reaction. The risk of serious adverse reactions to AX increased in older patients and in males, but this trend was not significant.


Metabolic Characterization of Antifolate Responsiveness and Non-responsiveness in Malignant Pleural Mesothelioma Cells.
Antifolates are a class of drugs effective for treating malignant pleural mesothelioma (MPM). The majority of antifolates inhibit enzymes involved in purine and pyrimidine synthesis such as dihydrofolate reductase (DHFR), thymidylate synthase (TYMS), and glycinamide ribonucleotide formyltransferase (GART). In order to select the most suitable patients for effective therapy with drugs targeting specific metabolic pathways, there is a need for better predictive metabolic biomarkers. Antifolates can alter global metabolic pathways in MPM cells, yet the metabolic profile of treated cells has not yet been clearly elucidated. Here we found that MPM cell lines could be categorized into two groups according to their sensitivity or resistance to pemetrexed treatment. We show that pemetrexed susceptibility could be reversed and DNA synthesis rescued in drug-treated cells by the exogenous addition of the nucleotide precursors hypoxanthine and thymidine (HT). We observed that the expression of pemetrexed-targeted enzymes in resistant MPM cells was quantitatively lower than that seen in pemetrexed-sensitive cells. Metabolomic analysis revealed that glycine and choline, which are involved in one-carbon metabolism, were altered after drug treatment in pemetrexed-sensitive but not resistant MPM cells. The addition of HT upregulated the concentration of inosine monophosphate (IMP) in pemetrexed-sensitive MPM cells, indicating that the nucleic acid biosynthesis pathway is important for predicting the efficacy of pemetrexed in MPM cells. Our data provide evidence that may link therapeutic response to the regulation of metabolism, and points to potential biomarkers for informing clinical decisions regarding the most effective therapies for patients with MPM.


Radiation exposure from abdominal imaging studies in patients with intestinal Behçet disease.
Recently, several studies have revealed that diagnostic imaging can result in exposure to harmful levels of ionizing radiation in inflammatory bowel disease patients. However, the extent of radiation exposure in intestinal Behcet disease (BD) patients has not been documented. The aim of this study was to estimate the radiation exposure from abdominal imaging studies in intestinal BD patients. Patients with a diagnosis of intestinal BD established between January 1990 and March 2012 were investigated at a single tertiary academic medical center. The cumulative effective dose (CED) was calculated retrospectively from standard tables and by counting the number of abdominal imaging studies performed. High exposure was defined as CED >50 mSv. In total, 270 patients were included in the study. The mean CED was 41.3 mSv, and 28.1% of patients were exposed to high levels of radiation. Computed tomography (CT) accounted for 81.7% of the total effective dose. In multivariate analyses, predictors of high radiation exposure were azathioprine/6-mercaptopurine use, surgery, and hospitalization. Approximately a quarter of intestinal BD patients were exposed to harmful levels of diagnostic radiation, mainly from CT examination. Clinicians should reduce the number of unnecessary CT examinations and consider low-dose CT profiles or alternative modalities such as magnetic resonance enterography.


Management of epileptic seizures in school-age children: Educational project dedicated to school staff.
The objective of this study was to educate the school staff for a correct management of epileptic seizures in order to increase the safety of young people at school and promoting the administration of rescue drugs and in order to improve care and reduce improper calls to the health emergency number. This project started in January 2016, and it is still ongoing at the Department of Neuroscience of Bambino Gesù Children's Hospital in Rome, Italy. There has been a data cut-off evaluation in November 2018. Two-hour training meetings with the school staff have been organized. The major topics of the training activities were as follows: report what epilepsy is, how to manage students with epileptic seizures, and how to administer rescue medications. During the meetings, the following two questionnaires were administered: one pretest in order to collect personal information and information on awareness of epilepsy, willingness to administer rescue medications, and anxiety in facing a seizure; and one posttest in order to check the knowledge acquired after the training sessions. Statistical analysis was performed using R version 3.2.3 (R Foundation for Statistical Computing, http://www.R-project.org/). Demographics (sex and age) and teaching experience were summarized with descriptive statistics for each variable. Demographics, teaching experience, awareness of disability, and knowledge of epilepsy were correlated to the management of seizures occurring in the classroom before the course; results are reported as odds ratios [OR] and 95% confidence interval (95 CI). Nine hundred school staff members (95% school staff and 5% social workers) entered in the project between January 2016 and November 2018. Seven hundred and forty (82%) returned the questionnaires fulfilled, and not all of them were completely filled. Ninety-eight percent of school staff (676/691) were aware about epilepsy; however, only in 16% (110) the awareness of epilepsy came from medical staff, scientific brochures, or participation in conventions. Thirty-five percent of school staff (248/707) believed that epilepsy reduces learning abilities, and 58% (409/703) believed that children with epilepsy need school support. After the training, 68% of school staff (496/734) correctly filled in the questionnaire related to the management of acute seizures versus 8% of them (57/718) in the prequestionnaire. After the training, 89% of school staff (601/675) were ready to administer rescue medications versus 54% (384/712) before the training. The majority of participants reported that the level of anxiety related to the management of seizures after the training significantly reduced. Results of this project documented an increase in knowledge of epilepsy, a better knowledge on management of acute seizures in the school settings, a reduction in anxiety, and an increase in willingness to administer rescue medications. Further studies should be planned in order to document the changes in the real-world management of seizures, to evaluate if a reduction in hospital admittances might be reached, and to extend the project by assessing, through a questionnaire, the stigma and prejudices against the children affected by epilepsy by their classmates.


Ventral hernia repair with poly-4-hydroxybutyrate mesh.
Biomaterial research has made available a biologically derived fully resorbable poly-4-hydroxybutyrate (P4HB) mesh for use in ventral and incisional hernia repair (VIHR). This study evaluates outcomes of patients undergoing VIHR with P4HB mesh. An IRB-approved prospective pilot study was conducted to assess clinical and quality of life (QOL) outcomes for patients undergoing VIHR with P4HB mesh. Perioperative characteristics were defined. Clinical outcomes, employment status, QOL using 12-item short form survey (SF-12), and pain assessments were followed for 24 months postoperatively. 31 patients underwent VIHR with bioresorbable mesh via a Rives-Stoppa approach with retrorectus mesh placement. The median patient age was 52 years, median body mass index was 33 kg/m2, and just over half of the patients were female. Surgical site occurrences occurred in 19% of patients, most of which were seroma. Hernia recurrence rate was 0% (median follow-up = 414 days). Patients had significantly improved QOL at 24 months compared to baseline for SF-12 physical component summary and role emotional (p < 0.05). Ventral hernia repair with P4HB bioresorbable mesh results in favorable outcomes. Early hernia recurrence was not identified among the patient cohort. Quality of life improvements were noted at 24 months versus baseline for this cohort of patients with bioresorbable mesh. Use of P4HB mesh for ventral hernia repair was found to be feasible in this patient population. (ClinicalTrials.gov Identifier: NCT01863030).


Osteoblasts in bone physiology-mini review.
Bone structural integrity and shape are maintained by removal of old matrix by osteoclasts and in-situ synthesis of new bone by osteoblasts. These cells comprise the basic multicellular unit (BMU). Bone mass maintenance is determined by the net anabolic activity of the BMU, when the matrix elaboration of the osteoblasts equals or exceeds the bone resorption by the osteoclasts. The normal function of the BMU causes a continuous remodeling process of the bone, with deposition of bony matrix (osteoid) along the vectors of the generated force by gravity and attached muscle activity. The osteoblasts are derived from mesenchymal stem cells (MSCs). Circulating hormones and locally produced cytokines and growth factors modulate the replication and differentiation of osteoclast and osteoblast progenitors. The appropriate number of the osteoblasts in the BMU is determined by the differentiation of the precursor bone-marrow stem cells into mature osteoblasts, their proliferation with subsequent maturation into metabolically active osteocytes, and osteoblast degradation by apoptosis. Thus, the two crucial points to target when planning to control the osteoblast population are the processes of cell proliferation and apoptosis, which are regulated by cellular hedgehog and Wnt pathways that involve humoral and mechanical stimulations. Osteoblasts regulate both bone matrix synthesis and mineralization directly by their own synthetic activities, and bone resorption indirectly by its paracrinic effects on osteoclasts. The overall synthetic and regulatory activities of osteoblasts govern bone tissue integrity and shape.


Diagnosis and treatment of autoimmune and IgG4-related hypophysitis: clinical guidelines of the Japan Endocrine Society.
Hypophysitis, which is often accompanied by pituitary dysfunction, is classified into several subtypes based on the cause, histology, and the location of inflammation in the pituitary gland. A definitive diagnosis requires pituitary biopsy, which is invasive, and the process is limited to specialized clinical settings. In this opinion paper, we review the literature associated with hypophysitis, and provide the guidelines of the Japan Endocrine Society for the diagnosis and treatment of autoimmune and IgG4-related hypophysitis.


Iron acquisition by Mycobacterium tuberculosis: isolation and characterization of a family of iron-binding exochelins.
Mycobacterium tuberculosis, the primary agent of tuberculosis, must acquire iron from the host to cause infection. To do so, it releases high-affinity iron-binding siderophores called exochelins. Exochelins are thought to transfer iron to another type of high-affinity iron-binding molecule in the bacterial cell wall, mycobactins, for subsequent utilization by the bacterium. In this paper, we describe the purification of exochelins of M. tuberculosis and their characterization by mass spectrometry. Exochelins comprise a family of molecules whose most abundant species range in mass from 744 to 800 Da in the neutral Fe(3+)-loaded state. The molecules form two 14-Da-increment series, one saturated and the other unsaturated, with the increments reflecting different numbers of CH2 groups on a side chain. These series further subdivide into serine- or threonine-containing species. The virulent M. tuberculosis Erdman strain and the avirulent M. tuberculosis H37Ra strain produce a similar set of exochelins. Based on a comparison of their tandem mass spectra, exochelins share a common core structure with mycobactins. However, exochelins are smaller than mycobactins due to a shorter alkyl side chain, and the side chain of exochelins terminates in a methyl ester. These differences render exochelins more polar than the lipophilic mycobactins and hence soluble in the aqueous extracellular milieu of the bacterium in which they bind iron in the host.


Methods to assess area and volume of wounds - a systematic review.
Wound measurement is important in monitoring the healing process of chronic wounds and in evaluating the effect of treatment. The objective of this systematic review was to evaluate evidence from the literature on accuracy, agreement, reliability and feasibility of wound measurement techniques described since 1994. Studies were identified by searching the electronic databases PubMed, Embase and Cochrane Library. Of the 12 013 studies identified, 43 were included in the review. A total of 30 papers evaluated techniques for measuring wound area and 13 evaluated techniques for measuring wound volume. The six approaches for measuring wound area were simple ruler method (10 papers), mathematical models (5 papers), manual planimetry (10 papers), digital planimetry (16 papers), stereophotogrammetry (2 papers) and digital imaging method (20 papers). Of these studies, 10 evaluated accuracy, 15 agreement, 17 reliability and 25 mentioned feasibility. The number of wounds examined in the studies was highly variable (n = 3-260). Studies evaluating techniques for measuring wound volume included between 1 and 50 wounds and evaluated accuracy (4 studies), agreement (6 studies), reliability (8 studies) and feasibility (12 studies). Digital planimetry and digital imaging were considered the most accurate and reliable methods for area measurement, particularly in larger and irregularly shaped wounds. None of the three-dimensional technologies have so far had a major impact, because of their low accuracy, high cost and complexity in handling the system set-up.


TRACE (Trunk Aesthetic Clinical Evaluation), a routine clinical tool to evaluate aesthetics in scoliosis patients: development from the Aesthetic Index (AI) and repeatability.
Aesthetic appearance is of primary importance in the treatment of adolescent idiopathic scoliosis (AIS), but to date tools for routine clinical practice have not become available. The aim of the present study is to develop such a tool and to verify its repeatability. At first we developed the Aesthetic Index (AI), based on a three-point scale for asymmetry of the shoulders, scapulae and waist that we tested for 5 years. From this experience we developed another tool we called TRACE, the acronym of Trunk Aesthetic Clinical Evaluation; TRACE is a 12-point scale based on four sub-scales, shoulders (0-3), scapulae (0-2), hemi-thorax (0-2) and waist (0-4). Posterior-anterior (PA) photographs of one hundred-sixty AIS patients. Each photograph was scored in two independent tests by four observers using AI, and subsequently TRACE. Kappa statistical analysis and 95% level of agreement were used; we also identified the minimum significant change (95% confidence level). We found the intra- and inter-raters repeatability of AI to be fair. Three points out of seven was the minimum significant change between two different evaluations. For TRACE, intra-rater repeatability was fair and inter-raters poor; but the minimum significant change was three (intra-rater), or four (inter-raters) out of twelve points. Widening the scale from 7 (AI) to 12 points (TRACE) increased the clinical sensitivity to changes of the aesthetic scale, even if TRACE has only a fair repeatability. TRACE is a no-cost tool for routine clinical practice in AIS patients. Due to the absence of other comparable validated tools, once the inherent measurement error is known and understood, its routine clinical use by physicians is advised.


Highly conserved nucleotide phosphatase essential for membrane lipid homeostasis in Streptococcus pneumoniae.
Proteins belonging to the DHH family, a member of the phosphoesterase superfamily, are produced by most bacterial species. While some of these proteins are well studied in Bacillus subtilis and Escherichia coli, their functions in Streptococcus pneumoniae remain unclear. Recently, the highly conserved DHH subfamily 1 protein PapP (SP1298) has been reported to play an important role in virulence. Here, we provide a plausible explanation for the attenuated virulence of the papP mutant. Recombinant PapP specifically hydrolyzed nucleotides 3'-phosphoadenosine-5'-phosphate (pAp) and 5'-phosphoadenylyl-(3'->5')-adenosine (pApA). Deletion of papP, potentially leading to pAp/pApA accumulation, resulted in morphological defects and mis-localization of several cell division proteins. Incubation with both polar solvent and detergent led to robust killing of the papP mutant, indicating that membrane integrity is strongly affected. This is in line with previous studies showing that pAp inhibits the ACP synthase, an essential enzyme involved in lipid precursor production. Remarkably, partial inactivation of the lipid biosynthesis pathway, by inhibition of FabF or depletion of FabH, phenocopied the papP mutant. We conclude that pAp and pApA phosphatase activity of PapP is required for maintenance of membrane lipid homeostasis providing an explanation how inactivation of this protein may attenuate pneumococcal virulence.


Microwave absorbing property optimization of starlike ZnO/reduced graphene oxide doped by ZnO nanocrystal composites.
This study constructs a unique structure of reduced graphene oxide (RGO)/ZnO composite combining starlike ZnO with RGO doped by ZnO nanocrystals through an in situ crystallization method. After mixing graphene oxide (GO) dispersion with Zn(NO3)2 and KOH solution, zinc ions absorbed on graphene oxide sheets transform ZnO nanocrystals, and the free zinc ions form starlike ZnO. After thermal treatment, RGO sheets doped by uniformly dispersed ZnO nanocrystals wrap the starlike ZnO or insert in the gap between them. Composites of different mass ratio of RGO to ZnO (9 : 1, 8 : 1, 5.5 : 1, and 2 : 1) have been achieved by modulating the quantity of Zn(NO3)2 and KOH. The dielectric and microwave absorption properties of the composites show dependence on the mass ratio of RGO to ZnO. The sample with the mass ratio of 8 : 1 shows the most prominent microwave absorption properties, with strong absorption (minimum reflection loss of -77.5 dB) and broad effective absorption bandwidth (6.9 GHz). The enhanced microwave absorption properties can be ascribed to the starlike ZnO, which hinder the agglomeration of RGO, as well as the ZnO nanocrystals, effectively improving the interface of the heterostructure and the impedance matching of RGO.


Cold and dry winter conditions are associated with greater SARS-CoV-2 transmission at regional level in western countries during the first epidemic wave.
Higher transmissibility of SARS-CoV-2 in cold and dry weather conditions has been hypothesized since the onset of the COVID-19 pandemic but the level of epidemiological evidence remains low. During the first wave of the pandemic, Spain, Italy, France, Portugal, Canada and USA presented an early spread, a heavy COVID-19 burden, and low initial public health response until lockdowns. In a context when testing was limited, we calculated the basic reproduction number (R0) in 63 regions from the growth in regional death counts. After adjusting for population density, early spread of the epidemic, and age structure, temperature and humidity were negatively associated with SARS-CoV-2 transmissibility. A reduction of mean absolute humidity by 1 g/m3 was associated with a 0.15-unit increase of R0. Below 10 °C, a temperature reduction of 1 °C was associated with a 0.16-unit increase of R0. Our results confirm a dependency of SARS-CoV-2 transmissibility to weather conditions in the absence of control measures during the first wave. The transition from summer to winter, corresponding to drop in temperature associated with an overall decrease in absolute humidity, likely contributed to the intensification of the second wave in north-west hemisphere countries. Non-pharmaceutical interventions must be adjusted to account for increased transmissibility in winter conditions.


What lies beneath? Diffusion EAP-based study of brain tissue microstructure.
Diffusion weighted magnetic resonance signals convey information about tissue microstructure and cytoarchitecture. In the last years, many models have been proposed for recovering the diffusion signal and extracting information to constitute new families of numerical indices. Two main categories of reconstruction models can be identified in diffusion magnetic resonance imaging (DMRI): ensemble average propagator (EAP) models and compartmental models. From both, descriptors can be derived for elucidating the underlying microstructural architecture. While compartmental models indices directly quantify the fraction of different cell compartments in each voxel, EAP-derived indices are only a derivative measure and the effect of the different microstructural configurations on the indices is still unclear. In this paper, we analyze three EAP indices calculated using the 3D Simple Harmonic Oscillator based Reconstruction and Estimation (3D-SHORE) model and estimate their changes with respect to the principal microstructural configurations. We take advantage of the state of the art simulations to quantify the variations of the indices with the simulation parameters. Analysis of in-vivo data correlates the EAP indices with the microstructural parameters obtained from the Neurite Orientation Dispersion and Density Imaging (NODDI) model as a pseudo ground truth for brain data. Results show that the EAP derived indices convey information on the tissue microstructure and that their combined values directly reflect the configuration of the different compartments in each voxel.


Dehydroepiandrosterone secretion in healthy older men and women: effects of testosterone and growth hormone administration in older men.
Aging is associated with diminished gonadal steroid and GH/IGF-I axis activity; whether these changes contribute to the parallel declines of dehydroepiandrosterone (DHEA) and DHEA sulfate (DHEAS) production is unknown, as are the effects of sex steroid and/or GH administration on DHEA and DHEAS production. Our objective was to evaluate morning DHEAS concentrations and nocturnal DHEA secretory dynamics in healthy older men and women, before and after chronic administration of sex steroid(s) alone, GH alone, sex steroid(s) combined with GH, or placebo alone. We compared nocturnal DHEA secretory dynamics (2000 h to 0800 h, sampling every 20 min, analyzed by multiparameter deconvolution and approximate entropy algorithms) in healthy older (65-88 yr) men (n = 68) and women (n = 36), both before and after 26 wk of administration of sex steroid(s) alone [testosterone (T) in men or estrogen/progesterone in women], GH alone, sex steroid(s) combined with GH, or placebo alone. Morning concentrations of DHEAS were lower; nocturnal DHEA pulsatile production rate, burst frequency, and amplitude were higher; and half-life was shorter in women (P < 0.05). Nocturnal integrated DHEA concentrations, total production rate, and approximate entropy did not differ significantly by sex. Because of small treatment group sizes in women, only hormone intervention results in men are presented. In men, T and T plus GH administration significantly decreased nocturnal integrated DHEA but not morning DHEAS concentrations. GH alone exerted no significant effects on nocturnal DHEA secretion or morning DHEAS. Spontaneous nocturnal DHEA secretion is sexually dimorphic in healthy older individuals, and T administration decreases nocturnal DHEA secretion in older men. The clinical significance of sex steroid modulation of DHEA secretion in older persons remains to be elucidated.


A redescription of Carrolla craddocki (Lepospondyli: Brachystelechidae) based on high-resolution CT, and the impacts of miniaturization and fossoriality on morphology.
Some recent morphological analyses have brought into question the monophyly of Lissamphibia (frogs, salamanders, and caecilians). In these analyses, brachystelechid "microsaurs" are found to be sister group to caecilians. To test this hypothesis, the holotype specimen of the brachystelechid Carrolla craddocki was submitted to high-resolution X-ray computed tomography to gain insight into the nature of the morphology supporting the potential relationship between brachystelechids and caecilians. This analysis enabled us to conduct a detailed description of the internal anatomy such as the braincase and otic capsule endocast (the first of its kind for a lepospondyl), and new information regarding the architecture of the skull. Our results suggest brachystelechid cranial morphology is strongly influenced by miniaturization (enlarged sensory organs, anterior placement of the jaw articulation, and combination of both reduced- and hyper-ossifications) and burrowing habits (co-ossified braincase with broad, sloping occipital surface, overlapping joints between skull roof bones, and well-ossified anterior braincase). Characteristics of brachystelechids that appear unrelated to size-reduction and burrowing are the diamond-shaped skull and possible pedicellate dentition. We provide a revised diagnosis for Carrolla and identify possible new characters within the anatomy of the braincase and inner ear. Several characters currently uniting caecilians and "microsaurs" are among those associated with either miniaturization or burrowing, demonstrating that future efforts should continue to focus on fine details of anatomy minimally affected by these influences to contribute to the resolution of the question of the origin of caecilians.


Evaluating the reproducibility of topography systems on spherical surfaces.
Newly devised software was used to compare the ability of the Topographic Modeling System-1 (Computed Anatomy, New York, NY) and the Corneal Analysis System (EyeSys Laboratories, Houston, Tex) to reproduce power measurements on spherical surfaces. Reproducibility results were compared for spheres of 40.00, 42.50, and 44.00 diopters. The program determines the absolute difference in corneal power at defined keratoscope positions for paired examinations of the same eye. Four examinations of each sphere were obtained with each instrument. Individual points were sampled at specific keratoscope locations at 30 degrees-semimeridional intervals. The program compared variability of measurements at four defined ranges of distance from the vertex normal: within 0.60 mm, 0.61 to 1.5 mm, 1.51 to 2.5 mm, and 2.5 mm or greater. The Corneal Analysis System showed significantly greater variability of readings obtained within 0.60 mm of the vertex normal for all three spheres (P = .001 by Duncan's multiple comparison procedure), whereas the Topographic Modeling System-1 showed equally consistent readings within 0.60 mm as it did between 0.61 and 1.5 mm from the vertex normal.


Mutations in CDC45, Encoding an Essential Component of the Pre-initiation Complex, Cause Meier-Gorlin Syndrome and Craniosynostosis.
DNA replication precisely duplicates the genome to ensure stable inheritance of genetic information. Impaired licensing of origins of replication during the G1 phase of the cell cycle has been implicated in Meier-Gorlin syndrome (MGS), a disorder defined by the triad of short stature, microtia, and a/hypoplastic patellae. Biallelic partial loss-of-function mutations in multiple components of the pre-replication complex (preRC; ORC1, ORC4, ORC6, CDT1, or CDC6) as well as de novo stabilizing mutations in the licensing inhibitor, GMNN, cause MGS. Here we report the identification of mutations in CDC45 in 15 affected individuals from 12 families with MGS and/or craniosynostosis. CDC45 encodes a component of both the pre-initiation (preIC) and CMG helicase complexes, required for initiation of DNA replication origin firing and ongoing DNA synthesis during S-phase itself, respectively, and hence is functionally distinct from previously identified MGS-associated genes. The phenotypes of affected individuals range from syndromic coronal craniosynostosis to severe growth restriction, fulfilling diagnostic criteria for Meier-Gorlin syndrome. All mutations identified were biallelic and included synonymous mutations altering splicing of physiological CDC45 transcripts, as well as amino acid substitutions expected to result in partial loss of function. Functionally, mutations reduce levels of full-length transcripts and protein in subject cells, consistent with partial loss of CDC45 function and a predicted limited rate of DNA replication and cell proliferation. Our findings therefore implicate the preIC as an additional protein complex involved in the etiology of MGS and connect the core cellular machinery of genome replication with growth, chondrogenesis, and cranial suture homeostasis.


Effectiveness and cost-effectiveness of height-screening programmes during the primary school years: a systematic review.
To determine the effectiveness and cost-effectiveness of height screening (of children aged 4 to 11) to identify height-related conditions. Systematic review and economic modelling. We included published and unpublished screening studies of any design, except case reports, conducted in any setting that measured children's height as part of a population-level assessment. Studies were identified by electronic database searches, contact with experts and from bibliographies of retrieved studies. Children aged between 4 and 11 years. Diagnostic yield of height-related conditions and change in quality of life, as measured by quality-adjusted life years (QALYs), for early versus late treatment of underlying conditions. Twelve studies described a height-screening programme and provided data on the diagnostic yield of newly diagnosed height-related conditions. Where reported, yield for growth-hormone deficiency (per 1000 children screened) ranged from 0.05 (1 in 20,000) to 0.62 (approximately 1 in 1500) and for Turner syndrome (per 1000 children screened) was between 0.02 (1 in 50,000) and 0.07 (approximately 1 in 14,000). As a secondary gain, children with other potentially treatable conditions were identified; diagnostic yields ranged from 0.22 to 1.84 per 1000 children screened. Three studies did not detect any new cases, but all of these studies had methodological limitations. Economic modelling suggested that height screening is associated with health improvements and is cost effective for a willingness to pay threshold of pound 30,000 per QALY. This review indicates the utility and acceptable cost-effectiveness of height screening arising from increased detection of height-related disorders and secondary pick-up of other undiagnosed conditions. Further research is needed to obtain more reliable data on quality of life gains and costs associated with early interventions for height-related conditions. The exact role of height-screening programmes in improving child health remains to be determined.


Multifocus optical-resolution photoacoustic microscopy using stimulated Raman scattering and chromatic aberration.
In this Letter, multifocus optical-resolution photoacoustic microscopy is demonstrated using wavelength tuning and chromatic aberration for depth scanning. Discrete focal zones at several depth locations were created by refocusing light from a polarization-maintaining single-mode fiber pumped by a nanosecond fiber laser. The fiber and laser parameters were chosen to take advantage of stimulated Raman scattering (SRS) in the fiber to create a multiwavelength output that could then be bandpass filtered. The collimator lens and objective lens are chosen to take advantage of chromatic aberration in which each generated SRS wavelength peak focuses at a slightly different depth. The maximum amplitude of photoacoustic signals is mapped to form C-scan images. Additionally, all wavelength peaks fired simultaneously offers improved depth-of-field structural imaging at the cost of slight degradation of mainlobe-to-sidelobe ratios. Wavelength-tuned depth scanning over more than 440 μm is demonstrated, significantly greater than the ~100 μm depth of field predicted from our focused Gaussian beams. The improved depth of focus could be valuable for structural imaging of microvascular morphology without the need for mechanical scanning in the depth direction.


Enhanced production and perception of musical pitch in tone language speakers.
Individuals differ markedly with respect to how well they can imitate pitch through singing and in their ability to perceive pitch differences. We explored whether the use of pitch in one's native language can account for some of the differences in these abilities. Results from two studies suggest that individuals whose native language is a tone language, in which pitch contributes to word meaning, are better able to imitate (through singing) and perceptually discriminate musical pitch. These findings support the view that language acquisition fine-tunes the processing of critical auditory dimensions in the speech signal and that this fine-tuning can be carried over into nonlinguistic domains.


Wortmannin inactivates phosphoinositide 3-kinase by covalent modification of Lys-802, a residue involved in the phosphate transfer reaction.
Wortmannin at nanomolar concentrations is a potent and specific inhibitor of phosphoinositide (PI) 3-kinase and has been used extensively to demonstrate the role of this enzyme in diverse signal transduction processes. At higher concentrations, wortmannin inhibits the ataxia telangiectasia gene (ATM)-related DNA-dependent protein kinase (DNA-PKcs). We report here the identification of the site of interaction of wortmannin on the catalytic subunit of PI 3-kinase, p110alpha. At physiological pH (6.5 to 8) wortmannin reacted specifically with p110alpha. Phosphatidylinositol-4,5-diphosphate, ATP, and ATP analogs [adenine and 5'-(4-fluorosulfonylbenzoyl)adenine] competed effectively with wortmannin, while substances containing nucleophilic amino acid side chain functions had no effect at the same concentrations. This suggests that the wortmannin target site is localized in proximity to the substrate-binding site and that residues involved in wortmannin binding have an increased nucleophilicity because of their protein environment. Proteolytic fragments of wortmannin-treated, recombinant p110alpha were mapped with anti-wortmannin and anti-p110alpha peptide antibodies, thus limiting the target site within a 10-kDa fragment, colocalizing with the ATP-binding site. Site-directed mutagenesis of all candidate residues within this region showed that only the conservative Lys-802-to-Arg mutation abolished wortmannin binding. Inhibition of PI 3-kinase occurs, therefore, by the formation of an enamine following the attack of Lys-802 on the furan ring (at C-20) of wortmannin. The Lys-802-to-Arg mutant was also unable to bind FSBA and was catalytically inactive in lipid and protein kinase assays, indicating a crucial role for Lys-802 in the phosphotransfer reaction. In contrast, an Arg-916-to-Pro mutation abolished the catalytic activity whereas covalent wortmannin binding remained intact. Our results provide the basis for the design of novel and specific inhibitors of an enzyme family, including PI kinases and ATM-related genes, that play a central role in many physiological processes.


Evolutionary innovation and diversification of carotenoid-based pigmentation in finches.
The ornaments used by animals to mediate social interactions are diverse, and by reconstructing their evolutionary pathways we can gain new insights into the mechanisms underlying ornamental innovation and variability. Here, we examine variation in plumage carotenoids among the true finches (Aves: Fringillidae) using biochemical and comparative phylogenetic analyses to reconstruct the evolutionary history of carotenoid states and evaluate competing models of carotenoid evolution. Our comparative analyses reveal that the most likely ancestor of finches used dietary carotenoids as yellow plumage colorants, and that the ability to metabolically modify dietary carotenoids into more complex pigments arose secondarily once finches began to use modified carotenoids to create red plumage. Following the evolutionary "innovation" that enabled modified red carotenoid pigments to be deposited as plumage colorants, many finch species subsequently modified carotenoid biochemical pathways to create yellow plumage. However, no reversions to dietary carotenoids were observed. The finding that ornaments and their underlying mechanisms may be operating under different selection regimes-where ornamental trait colors undergo frequent reversions (e.g., between red and yellow plumage) while carotenoid metabolization mechanisms are more conserved-supports a growing empirical framework suggesting different evolutionary patterns for ornaments and the mechanistic innovations that facilitate their diversification.


Minimising pain in farm animals: the 3S approach - 'Suppress, Substitute, Soothe'.
Recently, the French National Institute for Agricultural Research appointed an expert committee to review the issue of pain in food-producing farm animals. To minimise pain, the authors developed a '3S' approach accounting for 'Suppress, Substitute and Soothe' by analogy with the '3Rs' approach of 'Reduction, Refinement and Replacement' applied in the context of animal experimentation. Thus, when addressing the matter of pain, the following steps and solutions could be assessed, in the light of their feasibility (technical constraints, logistics and regulations), acceptability (societal and financial aspects) and availability. The first solution is to suppress any source of pain that brings no obvious advantage to the animals or the producers, as well as sources of pain for which potential benefits are largely exceeded by the negative effects. For instance, tail docking of cattle has recently been eliminated. Genetic selection on the basis of resistance criteria (as e.g. for lameness in cattle and poultry) or reduction of undesirable traits (e.g. boar taint in pigs) may also reduce painful conditions or procedures. The second solution is to substitute a technique causing pain by another less-painful method. For example, if dehorning cattle is unavoidable, it is preferable to perform it at a very young age, cauterising the horn bud. Animal management and constraint systems should be designed to reduce the risk for injury and bruising. Lastly, in situations where pain is known to be present, because of animal management procedures such as dehorning or castration, or because of pathology, for example lameness, systemic or local pharmacological treatments should be used to soothe pain. These treatments should take into account the duration of pain, which, in the case of some management procedures or diseases, may persist for longer periods. The administration of pain medication may require the intervention of veterinarians, but exemptions exist where breeders are allowed to use local anaesthesia (e.g. castration and dehorning in Switzerland). Extension of such exemptions, national or European legislation on pain management, or the introduction of animal welfare codes by retailers into their meat products may help further developments. In addition, veterinarians and farmers should be given the necessary tools and information to take into account animal pain in their management decisions.


Transmission of ranavirus between ectothermic vertebrate hosts.
Transmission is an essential process that contributes to the survival of pathogens. Ranaviruses are known to infect different classes of lower vertebrates including amphibians, fishes and reptiles. Differences in the likelihood of infection among ectothermic vertebrate hosts could explain the successful yearlong persistence of ranaviruses in aquatic environments. The goal of this study was to determine if transmission of a Frog Virus 3 (FV3)-like ranavirus was possible among three species from different ectothermic vertebrate classes: Cope's gray treefrog (Hyla chrysoscelis) larvae, mosquito fish (Gambusia affinis), and red-eared slider (Trachemys scripta elegans). We housed individuals previously exposed to the FV3-like ranavirus with naïve (unexposed) individuals in containers divided by plastic mesh screen to permit water flow between subjects. Our results showed that infected gray treefrog larvae were capable of transmitting ranavirus to naïve larval conspecifics and turtles (60% and 30% infection, respectively), but not to fish. Also, infected turtles and fish transmitted ranavirus to 50% and 10% of the naïve gray treefrog larvae, respectively. Nearly all infected amphibians experienced mortality, whereas infected turtles and fish did not die. Our results demonstrate that ranavirus can be transmitted through water among ectothermic vertebrate classes, which has not been reported previously. Moreover, fish and reptiles might serve as reservoirs for ranavirus given their ability to live with subclinical infections. Subclinical infections of ranavirus in fish and aquatic turtles could contribute to the pathogen's persistence, especially when highly susceptible hosts like amphibians are absent as a result of seasonal fluctuations in relative abundance.


Giant piezoelectricity on Si for hyperactive MEMS.
Microelectromechanical systems (MEMS) incorporating active piezoelectric layers offer integrated actuation, sensing, and transduction. The broad implementation of such active MEMS has long been constrained by the inability to integrate materials with giant piezoelectric response, such as Pb(Mg(1/3)Nb(2/3))O(3)-PbTiO(3) (PMN-PT). We synthesized high-quality PMN-PT epitaxial thin films on vicinal (001) Si wafers with the use of an epitaxial (001) SrTiO(3) template layer with superior piezoelectric coefficients (e(31,f) = -27 ± 3 coulombs per square meter) and figures of merit for piezoelectric energy-harvesting systems. We have incorporated these heterostructures into microcantilevers that are actuated with extremely low drive voltage due to thin-film piezoelectric properties that rival bulk PMN-PT single crystals. These epitaxial heterostructures exhibit very large electromechanical coupling for ultrasound medical imaging, microfluidic control, mechanical sensing, and energy harvesting.


Multiple shRNAs expressed by an inducible pol II promoter can knock down the expression of multiple target genes.
RNA interference (RNAi) has been increasingly used for reverse genetics. Both pol III and pol II promoters have been used to synthesize short hairpin RNA (shRNA) for knockdown of gene expression in mammalian cells and animals. Compared with gene knockout approaches, RNAi has the advantage of being simple, quick, and low cost. Here we describe a method that enhances this advantage where knockdown of expression of multiple genes in the same cells is required. A tetracycline-regulated pol II promoter construct allows the expression of up to three shRNA genes that have been cloned into introns of a transcript bearing green fluorescent protein (GFP) coding sequences. This method may be used to establish stable knockdown cell lines and may also prove useful for investigating gene-gene interactions in transgenic animals.


Initiation and duration of laryngeal closure during the pharyngeal swallow in post-stroke patients.
As a bolus enters the pharynx during the swallow, the airway is protected by laryngeal closure, a process characterized by approximation of the vocal folds plus approximation of the arytenoid cartilages to the base of the epiglottis. The purpose of this study was to measure initiation of laryngeal closure (ILC) and laryngeal closure duration (LCD) in three groups of subjects: (1) ten stroke patients who aspirated before and during the swallow (aspirators), (2) ten stroke patients who did not aspirate (nonaspirators), and (3) ten normal control subjects. Means and standard deviations of ILC and LCD were analyzed for both 5-ml and 10-ml thin-liquid boluses using a 100-ms timer during subsequent analysis of videofluoroscopic swallowing examinations. There were significant differences between aspirators and control subjects for both ILC and LCD, and significant differences between aspirators and nonaspirators for ILC. There were no significant differences between aspirators and nonaspirators for LCD. Both delayed ILC and reduced LCD were associated with post-stroke aspiration. Delayed ILC is a significant indicator of overall risk of aspiration. Clinical implications for these findings are discussed.


Immune-Related Adverse Events and Immune Checkpoint Inhibitor Efficacy in Patients with Gastrointestinal Cancer with Food and Drug Administration-Approved Indications for Immunotherapy.
Immune-related adverse event (IRAE) onset may represent a clinical biomarker for anti-programmed cell death protein 1 (PD-1) antibody response based on emerging evidence from patients with various advanced malignancies. This phenomenon has not been previously reported in a multidisease cohort of patients with gastrointestinal (GI) cancer with Food and Drug Administration (FDA)-approved indications to receive immune checkpoint inhibitor therapy. The study was a multicenter retrospective cohort analysis of 76 patients with GI cancer who had received anti-PD-1 antibodies for FDA-approved indications. The primary and secondary outcomes of the study were progression-free survival (PFS) and overall survival (OS) in patients based upon IRAE presence, respectively. PFS and OS were estimated by the Kaplan-Meier method; a Cox proportional-hazards model adjusted for IRAE onset, patient age, and enrolling institution was used to analyze outcomes. Median PFS and OS were prolonged in patients who experienced IRAEs compared with those who did not experience them (PFS: not reached [NR] vs. 3.9 months [hazard ratio (HR) 0.13, 95% confidence interval (CI) 0.05-0.3, p < .001]; OS: NR vs. 7.4 months [HR 0.11, 95% CI 0.03-0.36, p < .001]). Among patients who experienced IRAEs, there were no significant differences in PFS and OS by either initial IRAE severity, management, or time to onset. Patients with gastrointestinal cancer who experienced IRAEs while on anti-PD-1 antibodies demonstrated significant improvements in PFS and OS compared with their counterparts who did not develop IRAEs. Although these findings add to results from studies in other tumor types, larger prospective studies are needed prior to clinical adoption of IRAE onset as a biomarker for immune checkpoint inhibitor response. Predictive clinical biomarkers for immune checkpoint inhibitor response have been understudied in the field of immuno-oncology. Immune-related adverse event onset appears to be one such biomarker. Across tumor types, immune-related adverse event onset has been associated with response to anti-programmed cell death protein 1 (PD-1) antibodies. The results of this study demonstrate this for the first time in patients with gastrointestinal cancer receiving anti-PD-1 antibodies. Before immune-related adverse event onset can be adopted clinically as a predictive biomarker for immune checkpoint inhibitor response, however, larger prospective studies are needed to better understand the nuances between immune-related adverse event characteristics (severity, site, management, timing of onset) and immune checkpoint inhibitor effectiveness.


Anti-NMDA Receptor Encephalitis and Vaccination.
Anti-N-methyl-d-aspartate (Anti-NMDA) receptor encephalitis is an acute autoimmune neurological disorder. The cause of this disease is often unknown, and previous studies revealed that it might be caused by a virus, vaccine or tumor. It occurs more often in females than in males. Several cases were reported to be related to vaccination such as the H1N1 vaccine and tetanus/diphtheria/pertussis and polio vaccines. In this study, we reported an anti-NMDA receptor encephalitis case that may be caused by Japanese encephalitis vaccination. To investigate the association between anti-NMDA receptor encephalitis and vaccination, we analyzed the phylogenetic relationship of the microRNAs, which significantly regulate these vaccine viruses or bacteria, and the phylogenetic relationship of these viruses and bacteria. This reveals that anti-NMDA receptor encephalitis may be caused by Japanese encephalitis vaccination, as well as H1N1 vaccination or tetanus/diphtheria/pertussis and polio vaccinations, from the phylogenetic viewpoint.


Coding of Electronic Laboratory Reports for Biosurveillance, Selected United States Hospitals, 2011.
Objective Electronic laboratory reporting has been promoted as a public health priority. The Office of the U.S. National Coordinator for Health Information Technology has endorsed two coding systems: Logical Observation Identifiers Names and Codes (LOINC) for laboratory test orders and Systemized Nomenclature of Medicine-Clinical Terms (SNOMED CT) for test results. Materials and Methods We examined LOINC and SNOMED CT code use in electronic laboratory data reported in 2011 by 63 non-federal hospitals to BioSense electronic syndromic surveillance system. We analyzed the frequencies, characteristics, and code concepts of test orders and results. Results A total of 14,028,774 laboratory test orders or results were reported. No test orders used SNOMED CT codes. To describe test orders, 77% used a LOINC code, 17% had no value, and 6% had a non-informative value, "OTH". Thirty-three percent (33%) of test results had missing or non-informative codes. For test results with at least one informative value, 91.8% had only LOINC codes, 0.7% had only SNOMED codes, and 7.4% had both. Of 108 SNOMED CT codes reported without LOINC codes, 45% could be matched to at least one LOINC code. Conclusion Missing or non-informative codes comprised almost a quarter of laboratory test orders and a third of test results reported to BioSense by non-federal hospitals. Use of LOINC codes for laboratory test results was more common than use of SNOMED CT. Complete and standardized coding could improve the usefulness of laboratory data for public health surveillance and response.


The placenta is simply a neuroendocrine parasite.
This paper will document the early scientific observations that kindled my neuroendocrinological interest in pre-eclampsia, a life-threatening disease that affects both mother and baby. My interest in this subject started with the placental origin of melanotrophin activity, moving on, through corticotrophin-releasing factor and its binding protein, to a tachykinin modified specifically in the placenta by phosphocholine, a post-translational moiety normally used by parasites to avoid immune surveillance and rejection. This work may finally have led to an understanding of the identity of the elusive placental factor that, whilst attempting to compensate for the poor implantation of the placenta, causes the many symptoms seen in the mother during pre-eclampsia.


Genetic heterogeneity among three Adi tribes of Arunachal Pradesh, India.
We studied the distribution of ABO blood groups among three little known subtribes of the Adi tribe, namely, the Panggi, Komkar, and Padam, of the East and Upper Siang districts of Arunachal Pradesh, India. Blood group O was the predominant group in the Komkar and Padam, whereas group A was the predominant group in the Panggi. Blood group AB was found to be the least frequent group in all three studied populations. The populations showed significant differences in blood groups A (43% in Panggi, 23% in Komkar, and 18% in Padam) and O (33% in Panggi, 54% in Komkar, and 61% in Padam). The chi-square test indicated significant deviation from Hardy-Weinberg equilibrium, suggesting high heterogeneity among the tribes.