Stability of biomarkers of oxidative stress in canine serum.
The purpose of this study was to determine the effect of storage time and temperature on the measured concentrations of a profile of biomarkers of oxidative stress in canine serum: Trolox equivalent antioxidant capacity (TEAC) measured by two different methods, using acidic medium (TEACA), and horseradish peroxidase (TEACH), cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of plasma (FRAP), total thiol, Paraoxonase 1 (PON1), total oxidant status (TOS), ferrous oxidation xylenol orange (FOX), advanced oxidation protein products (AOPP), reactive oxygen species (ROS), and thiobarbituric reactive species (TBARS). Six serum pools from healthy dogs were divided in various aliquots and stored at 4 °C and 25 °C for a period of 6, 24 and 72 h, and at -20 °C and - 80 °C for a period of 7, 14, 30, 60, 180, and 360 days. In addition, the effect of repeated freeze-thaw cycle was evaluated. For the short-term storage of biomarkers of oxidative stress in canine serum, 4 °C is preferred to room temperature. In case of long-term storage, the optimal stability was achieved when samples were stored at -80 °C. Most of the biomarkers were affected after two freeze-thaw cycles. In addition, as individual oxidative biomarkers can show differences in stability, it is important to consider the stability of each different analyte when it is measured in stored samples and avoid the use of samples stored in conditions in which the analyte is not stable.


Retinoic acid accelerates downregulation of the Xist repressor, Oct4, and increases the likelihood of Xist activation when Tsix is deficient.
Imbalances in X-linked gene dosage between the sexes are resolved by transcriptionally silencing one of two X-chromosomes in female cells of the early mammalian embryo. X-inactivation is triggered by expression of the non-coding Xist gene. In turn, Xist is dually regulated by the antisense Tsix RNA and by the Oct4 pluripotency factor. Although there is general agreement that Tsix is an inhibitor of Xist, some laboratories have observed ectopic Xist induction in differentiating male ES cells when Tsix is mutated, whereas we have not observed significant changes in Xist. These observational differences have led to fundamentally diverse models of X-chromosome counting. Here, we investigate if different methods of cell differentiation and use of all -trans retinoic acid (RA) could be causative factors and how they might impact Xist expression. We compared suspension and cell-adhesion cultures in the presence or absence of RA and find that RA significantly impacts Xist expression in Tsix-mutant male cells. Whereas the standard embryoid body method infrequently leads to ectopic Xist expression, adding RA generates a significant number of Xist-positive male cells. However, while normal Xist clouds in wild-type female cells are robust and well-circumscribed, those found in the RA-treated mutant males are loosely dispersed. Furthermore, ectopic Xist expression does not generally lead to complete gene silencing. We attribute the effect of RA on Xist to RA's repressive influence on Oct4, a pluripotency factor recently shown to regulate Tsix and Xist. RA-treated ES cells exhibit accelerated decreases in Oct4 RNA levels and also display accelerated loss of binding to Xist intron 1. When Tsix is deficient, the faster kinetics of Oct4 loss tip the equilibrium towards Xist expression. However, the aberrant Xist clusters are unlikely to explain elevated cell death, as X-linked silencing does not necessarily correlate with the qualitatively aberrant Xist clusters. We conclude that RA treatment leads to premature downregulation of Oct4 and partial derepression of Xist irrespective of X-chromosome counting. RA-induced Xist clusters in male cells do not result in global or stable silencing, and excess cell death is not observed. These data and RA's known pleiotropic effects on ES transcription networks suggest that RA differentation bypasses normal X-inactivation controls and should be used judiciously. We propose that the likelihood of Xist expression is determined by a balance of multiple Xist activators and repressors, and that levels of Oct4 and Tsix are crucial toward achieving this balance.


Effects of tumor necrosis factor alpha on proliferation and expression of differentiated phenotypes in rabbit costal chondrocytes in culture.
Tumor necrosis factor alpha (TNF alpha) decreased the synthesis of glycosaminoglycan (GAG) in rabbit costal chondrocytes in culture, but did not stimulate the release of GAG from cell layers. Like chondrocytes cultured in control medium, chondrocytes cultured in the presence of TNF alpha produced putative "cartilage-specific" proteoglycans identified by density gradient centrifugation under dissociative conditions. Although TNF alpha decreased the synthesis of the proteoglycans, it did not change their monomeric size, which is a marker of cartilage phenotypes. Moreover, TNF alpha did not affect the responsiveness to parathyroid hormone, insulin-like growth factor I, or transforming growth factor beta, which is known to stimulate GAG synthesis in cultured chondrocytes. TNF alpha decreased the alkaline phosphatase activity in the chondrocytes dose dependently. On the other hand, it stimulated their DNA synthesis slightly, but significantly. The stimulatory effect of TNF alpha on DNA synthesis was potentiated by fibroblast growth factor, epidermal growth factor, and fetal bovine serum. These findings suggest that in the presence of hormones and growth factors, TNF alpha promotes the proliferation of chondrocytes while suppressing their further differentiation at the stage of synthesis of cartilage-specific proteoglycans.


The first mecp2-null zebrafish model shows altered motor behaviors.
Rett syndrome (RTT) is an X-linked neurodevelopmental disorder and one of the most common causes of mental retardation in affected girls. Other symptoms include a rapid regression of motor and cognitive skills after an apparently early normal development. Sporadic mutations in the transcription factor MECP2 has been shown to be present in more than 90% of the patients and several models of MeCP2-deficient mice have been created to understand the role of this gene. These models have pointed toward alterations in the maintenance of the central nervous system rather than its development, in line with the late onset of the disease in humans. However, the exact functions of MeCP2 remain difficult to delineate and the animal models have yielded contradictory results. Here, we present the first mecp2-null allele mutation zebrafish model. Surprisingly and in contrast to MeCP2-null mouse models, mecp2-null zebrafish are viable and fertile. They present nonetheless clear behavioral alterations during their early development, including spontaneous and sensory-evoked motor anomalies, as well as defective thigmotaxis.


Pelvic and breast examination skills curricula in United States medical schools: a survey of obstetrics and gynecology clerkship directors.
Learning to perform pelvic and breast examinations produces anxiety for many medical students. Clerkship directors have long sought strategies to help students become comfortable with the sensitive nature of these examinations. Incorporating standardized patients, simulation and gynecologic teaching associates (GTAs) are approaches gaining widespread use. However, there is a paucity of literature guiding optimal approach and timing. Our primary objective was to survey obstetrics and gynecology (Ob/Gyn) clerkship directors regarding timing and methods for teaching and assessment of pelvic and breast examination skills in United States medical school curricula, and to assess clerkship director satisfaction with current educational strategies at their institutions. Ob/Gyn clerkship directors from all 135 Liaison Committee on Medical Education accredited allopathic United States medical schools were invited to complete an anonymous 15-item web-based questionnaire. The response rate was 70%. Pelvic and breast examinations are most commonly taught during the second and third years of medical school. Pelvic examinations are primarily taught during the Ob/Gyn and Family Medicine (FM) clerkships, while breast examinations are taught during the Ob/Gyn, Surgery and FM clerkships. GTAs teach pelvic and breast examinations at 72 and 65% of schools, respectively. Over 60% of schools use some type of simulation to teach examination skills. Direct observation by Ob/Gyn faculty is used to evaluate pelvic exam skills at 87% of schools and breast exam skills at 80% of schools. Only 40% of Ob/Gyn clerkship directors rated pelvic examination training as excellent, while 18% rated breast examination training as excellent. Pelvic and breast examinations are most commonly taught during the Ob/Gyn clerkship using GTAs, simulation trainers and clinical patients, and are assessed by direct faculty observation during the Ob/Gyn clerkship. While the majority of Ob/Gyn clerkship directors were not highly satisfied with either pelvic or breast examination training programs, they were less likely to describe their breast examination training programs as excellent as compared to pelvic examination training-overall suggesting an opportunity for improvement. The survey results will be useful in identifying future challenges in teaching such skills in a cost-effective manner.


"It's about having money, but also happiness:" A qualitative investigation of how adolescents understand subjective status in themselves and others.
Understanding why adolescent subjective assessments of status matter to their psychosocial outcomes over and above objective assessments of socioeconomic status (SES) requires a better comprehension of how adolescents construct status in themselves and others. Using a qualitative approach, the goal of the current study was to better understand what factors adolescents use to assign status, and how their perceptions of this status vary according to their own SES. Qualitative interviews were conducted with 32 adolescents from Quebec, Canada, from varied socioeconomic backgrounds. In particular, they were shown the ladder from the Subjective Social Status Scale and asked what defined scale placement in their communities. They were also asked where they would place themselves on this scale, and why. The importance of traditional indicators of SES such as money, educational level, and occupational prestige were underlined by almost all adolescents, but other factors (e.g., wellbeing, family life, values concerning work, other-orientation, or rule-following) were also frequently discussed. Adolescents used similar criteria in the placement of themselves and others. Contradictions often emerged, however, as adolescents' scale placement was often higher than what would be expected based on traditional SES indicators. These findings provide a starting point for understanding why adolescents' subjective assessments of status may matter above and beyond traditional assessments of SES, and how we can better measure this construct.


RU 58841, a new specific topical antiandrogen: a candidate of choice for the treatment of acne, androgenetic alopecia and hirsutism.
A new topically active non-steroidal antiandrogen, RU 58841 has been synthesized. It displays high affinity for the hamster prostate and flank organ (F.O.) androgen receptors. In vivo, when topically applied, it exerts a potent dose-dependent regression of F.O. area at a dose as low as 1 microgram/animal while being devoid of antiandrogenic activity on deep accessory sex organs and of any effect on testosterone level up to 100 micrograms/animal. In the same species, after subcutaneous administration, it induces at the dose of 300 micrograms/animal, a small decrease in F.O. area equivalent to that of 1 microgram applied topically and a weak systemic activity. In intact rats, no effects were observed up to 1 microgram/animal whatever the route of administration. These results suggest that RU 58841 might useful for the topical treatment of androgen-dependent skin disorders such as acne, androgenetic alopecia and hirsutism.


Targeting Brd4 for cancer therapy: inhibitors and degraders.
Bromodomain-containing protein 4 (Brd4) plays an important role in mediating the expression of genes involved in cancers and non-cancer diseases such as inflammatory diseases and acute heart failure. Inactivating Brd4 or downregulating its expression inhibits cancer development, leading to the current interest in Brd4 as a promising anticancer drug target. Numerous Brd4 inhibitors have been studied in recent years and some of them are currently in various phases of clinical trials. Recently, selective degradation of target proteins by small bifunctional molecules (PROTACs) has emerged as an attractive drug discovery approach owing to the advantages it could offer over traditional small-molecule inhibitors. A number of Brd4 degraders have been reported and showed more efficient anticancer activities than just protein inhibition. In this review, we will discuss recent findings in the discovery and development of small-molecule inhibitors and degraders that target Brd4 as a potential anticancer agent.


Chronic cerebroventricular infusion of hypertonic sodium chloride in rats reduces hypothalamic sympatho-inhibition and elevates blood pressure.
To determine whether or not salt loading restricted to the brain would elevate blood pressure, chronically implanted osmotic minipumps were used to infuse hypertonic sodium chloride solutions made in artificial cerebrospinal fluid into a jugular vein intravenously or the third cerebral ventricle, intracerebroventricularly, for 11 days in awake rats. During intracerebroventricular infusion of hypertonic sodium chloride, tail-cuff systolic pressures began to rise on day 4 and were significantly elevated by day 9. In contrast, infusions of either artificial cerebrospinal fluid alone, intracerebroventricularly, or hypertonic sodium chloride, intravenously, were ineffective. Confirming the blood pressure elevation thereby detected, systolic and mean pressures recorded directly from indwelling aortic catheters after the same rats had been anesthetized with urethane on day 11, were also significantly higher following intracerebroventricular infusion of hypertonic sodium chloride than after infusion of artificial cerebrospinal fluid alone. Magnitude of depressor and sympatho-inhibitory responses elicited by graded electrical stimulation of the anterior hypothalamus invariably increased with the current strength used for stimulation. At all current strengths used for hypothalamic stimulation, depressor responses as well as attendant decreases in sympathetic neural firing, were smaller in rats that had been infused intracerebroventricularly with hypertonic sodium chloride than in any of the controls. Inhibition seemed specific for the anterior hypothalamus because pressor responses to stimulation of the ventromedial hypothalamus were the same whether or not the intracerebroventricular infusion contained hypertonic sodium chloride. An explanation based on diminished cardiovascular sensitivity also appeared unlikely, since depressor responses to intravenously injected histamine were almost equal in both groups. Our results are compatible with the interpretation that hypertonic sodium chloride infused chronically into the 3rd ventricle acts by reducing anterior hypothalamic inhibition of sympathetic vasomotor tone, and that this, in turn, then elevates blood pressure.


Nanopore sequencing detects structural variants in cancer.
Despite advances in sequencing, structural variants (SVs) remain difficult to reliably detect due to the short read length (<300 bp) of 2nd generation sequencing. Not only do the reads (or paired-end reads) need to straddle a breakpoint, but repetitive elements often lead to ambiguities in the alignment of short reads. We propose to use the long-reads (up to 20 kb) possible with 3rd generation sequencing, specifically nanopore sequencing on the MinION. Nanopore sequencing relies on a similar concept to a Coulter counter, reading the DNA sequence from the change in electrical current resulting from a DNA strand being forced through a nanometer-sized pore embedded in a membrane. Though nanopore sequencing currently has a relatively high mismatch rate that precludes base substitution and small frameshift mutation detection, its accuracy is sufficient for SV detection because of its long reads. In fact, long reads in some cases may improve SV detection efficiency. We have tested nanopore sequencing to detect a series of well-characterized SVs, including large deletions, inversions, and translocations that inactivate the CDKN2A/p16 and SMAD4/DPC4 tumor suppressor genes in pancreatic cancer. Using PCR amplicon mixes, we have demonstrated that nanopore sequencing can detect large deletions, translocations and inversions at dilutions as low as 1:100, with as few as 500 reads per sample. Given the speed, small footprint, and low capital cost, nanopore sequencing could become the ideal tool for the low-level detection of cancer-associated SVs needed for molecular relapse, early detection, or therapeutic monitoring.


Heterometallic molecular precursors for a lithium-iron oxide material: synthesis, solid state structure, solution and gas-phase behaviour, and thermal decomposition.
Three heterometallic single-source precursors with a Li : Fe = 1 : 1 ratio for a LiFeO2 oxide material are reported. Heterometallic compounds LiFeL3 (L = tbaoac (1), ptac (2), and acac(3)) have been obtained on a large scale, in nearly quantitative yields by one-step reactions that employ readily available reagents. The heterometallic precursor LiFe(acac)3 (3) with small, symmetric substituents on the ligand (acac = pentane-2,4-dionate), maintains a 1D polymeric structure in the solid state that limits its volatility and prevents solubility in non-coordinating solvents. The application of the unsymmetrical ligands, tbaoac (tert-butyl acetoacetate) and ptac (1,1,1-trifluoro-5,5-dimethyl-2,4-hexanedionate), that exhibit different bridging properties at the two ends of the ligand, allowed us to change the connectivity pattern within the heterometallic assembly. The latter was demonstrated by structural characterization of heterometallic complexes LiFe(tbaoac)3 (1) and LiFe(ptac)3 (2) that consist of discrete heterocyclic tetranuclear molecules Li2Fe2L6. The compounds are highly volatile and exhibit a congruent sublimation character. DART mass spectrometric investigation revealed the presence of heterometallic molecules in the gas phase. The positive mode spectra are dominated by the presence of [M - L]+ peaks (M = Li2Fe2L6). In accord with their discrete molecular structure, complexes 1 and 2 are highly soluble in nearly all common solvents. In order to test the retention of the heterometallic structure in solution, the diamagnetic analog of 1, LiMg(tbaoac)3 (4), has been isolated. Its tetranuclear molecular structure was found to be isomorphous to that of the iron counterpart. 1H and 7Li NMR spectroscopy unambiguously confirmed the presence of heterometallic molecules in solutions of non-coordinating solvents. The heterometallic precursor 1 was shown to exhibit clean thermal decomposition in air that results in phase-pure α-modification of layered oxide LiFeO2, the prospective cathode material for lithium ion batteries.


Polar opposites: Fine-tuning cytokinesis through SIN asymmetry.
Mitotic exit and cell division must be spatially and temporally integrated to facilitate equal division of genetic material between daughter cells. In the fission yeast, Schizosaccharomyces pombe, a spindle pole body (SPB) localized signaling cascade termed the septation initiation network (SIN) couples mitotic exit with cytokinesis. The SIN is controlled at many levels to ensure that cytokinesis is executed once per cell cycle and only after cells segregate their DNA. An interesting facet of the SIN is that its activity is asymmetric on the two SPBs during anaphase; however, how and why the SIN is asymmetric has remained elusive. Many key factors controlling SIN asymmetry have now been identified, shedding light on the significance of SIN asymmetry in regulating cytokinesis. In this review, we highlight recent advances in our understanding of SIN regulation, with an emphasis on how SIN asymmetry is achieved and how this aspect of SIN regulation fine-tunes cytokinesis.


Pediatricians' role in providing mental health care for children and adolescents: do pediatricians and child and adolescent psychiatrists agree?
Many children who have a mental health disorder do not receive mental health services and are seen only in primary care settings. Perceptions of pediatricians and mental health specialists regarding the role that pediatricians should have in diagnosing and managing children's mental health problems have not been studied. To examine whether primary care pediatricians (PCPs) and child and adolescent psychiatrists (CAPs) agree about: (1) the pediatrician's role in identification, referral, and treatment of childhood mental health (MH) disorders; and (2) barriers to the identification, referral, and treatment of childhood MH disorders. Surveys were mailed in 2005 to 338 PCPs and 75 CAPs in 7 counties surrounding Cleveland, Ohio. Each group was asked whether they agreed that PCPs should be responsible for identifying, treating, or referring 7 prevalent childhood MH problems. Barriers that PCPs face in identification, referral, and treatment of MH problems were also assessed. Analyses were weighted for nonresponse; group differences were assessed via Rao-Scott chi test and weighted regression analyses. Approximately half of PCPs and CAPs returned the survey. With the exception of attention deficit hyperactivity disorder (ADHD), the majority of PCPs and CAPs agreed that pediatricians should be responsible for identifying and referring, but not treating child MH conditions. For ADHD, PCPs were more likely than CAPs to agree that pediatricians should identify and treat affected children. PCPs were more likely than CAPs to agree that pediatricians should be responsible for identifying child/adolescent depression and anxiety disorders; the majority of both groups agree that PCPs should be responsible for referring, but not treating these conditions. Both groups agree that lack of MH services is a barrier to identification, treatment, and referral of child MH problems for PCPs. CAPs were more likely to agree that pediatrician's lack of training in identifying child mental health problems was a barrier, whereas PCPs were more likely to endorse lack of confidence in their ability to treat child MH problems with counseling, long waiting periods to see MH providers, family failure to follow through on referrals, and billing/reimbursement issues as barriers. Most PCPs and CAPs believe it is pediatricians' responsibility to identify and refer, but not treat, the majority of children's mental health problems. Both groups agree that mental health services are not readily available. Future efforts are needed to support PCPs and CAPs in their combined effort to address the mental health needs of children.


The persistence of clinical questions across shifts on an intensive care unit: an observational pilot study.
How do clinical questions emerge and move toward resolution in the intensive care setting over the course of 24 hours? In a 20-bed surgical intensive care unit in a large, tertiary-care teaching hospital, informationists shadowed clinicians for 2 48-hour periods to record questions, noting when they were asked and whether they were answered. Thirty-eight percent of 112 recorded questions remained unanswered. Some unanswered questions persisted across shifts, and clinicians' information-seeking behaviors changed over time. Clinical information services can be improved and integrated more fully into clinical workflows based on a fuller understanding of the life cycle of clinical inquiry.


Epidemiology of thoracolumbar spine injury in blunt trauma.
To evaluate the prevalence, distribution, and demographics of thoracolumbar (TL) spine injuries following blunt trauma. Prospective, cross-sectional study of a consecutive sample of all blunt trauma patients presenting initially to the emergency department (ED) of a Level 1 trauma center and undergoing thoracic and/or lumbar spine radiography from August 1997 to November 1998. The age, sex, and mechanism of injury of each patient as well as location and type of spine injury were recorded for those patients with vertebral fractures, dislocations, or subluxations. Two thousand four hundred four blunt trauma patients were enrolled. Vertebral injuries were identified in 152 individuals (6.3%, 95% CI = 5.4% to 7.4%). Two hundred sixty distinct anatomic levels of injury were identified in these 152 individuals. Of these 260 injuries, 42 (16.2%) occurred at L1, 38 (14.6%) at L2, 29 (11.1%) at L3, and 27 (10.4%) at T12, making these the most commonly injured vertebrae. Injuries were most common (34 patients) in those aged 30-39 years and were least common (12 patients) in those under 18 years. Compression fractures (52%) were the most common injury in the thoracic spine, while transverse process fractures (48%) were the most common injuries in the lumbar spine. The prevalence of TL injuries in ED blunt trauma patients undergoing TL radiographs is 6.3%. The most commonly injured area of the TL spine is the thoracolumbar junction.


Dental extraction in patients receiving dual antiplatelet therapy.
Dual antiplatelet therapy consists of administering antiplatelet (antiaggregant) drugs (clopidogrel and aspirin) to prevent thrombotic processes, as a preventative measure in patients with acute coronary disease, or in patients subjected to percutaneous coronary intervention. The purpose of this study was to evaluate the efficacy of a protocol for performing dental extraction in patients receiving dual antiplatelet therapy. Thirty-two patients undergoing dental extractions were included in the study. The variables evaluated were: collagen-epinephrine fraction, collagen- adenosine diphosphate fraction, surgical surface, post-surgical measures, and adverse effects. Alveolar sutures and gauzes impregnated with an antifibrinolytic agent (tranexamic acid), which the patient pressed in place for 30 minutes, were applied to all patients as post-surgical measures. Descriptive statistics were calculated and analyzed with Student's t-test to compare pairs of quantitative variables; simple regression analysis was performed using Pearson's correlation coefficient. Statistical significance was set at p<0.05. Collagen/epinephrine fraction was 264.53±55.624 seconds with a range of 135 to 300 seconds, and collagen/ADP fraction was 119.41±44.216 seconds, both values being higher than normal. As a result of the post-surgical measures taken, no patients presented post-operative bleeding, hematoma or infection. Dental extraction was safe for patients receiving dual anti-platelet therapy when using sutures and gauze impregnated with tranexamic acid, which the patient pressed in place for 30 minutes.


Spatial control of high-harmonic generation in hollow fibers.
We demonstrate the control of high-harmonic generation in a hollow fiber by shaping the spatial structure of the generating laser pulse. We use a liquid-crystal-based two-dimensional spatial light modulator to control the spatial phase of the driver pulse. An evolutionary algorithm finds the spatial laser phase distribution that is optimal for reaching maximum total harmonic yield and for selectively enhancing the cutoff region of the spectrum. We show that enhacement of harmonic generation is related to coupling into a single fiber mode. Our results directly show that spatial properties of the laser are important parameters in fully controlling the high-harmonic spectrum. It is thus not possible to derive the controllability of the high-harmonic generation from the single-atom response only.


Fourth Annual Pediatric Interagency Registry for Mechanical Circulatory Support (Pedimacs) Report.
The Pediatric Interagency Registry for Mechanical Circulatory Support (Pedimacs), originally a National Institutes of Health-sponsored U.S. database, provides a platform to understand the population of children supported with ventricular assist devices (VADs) during this time of increasing numbers, new devices, expanding indications, and improved outcomes. Between September 19, 2012, and December 31, 2019, 44 hospitals implanted 1031 devices in 856 patients under 19 years of age. Overall, diagnosis was cardiomyopathy in 497 (58%) patients, congenital heart disease (CHD) in 216 (25%), myocarditis in 85 (10%), and other in 58 (7%). Positive outcome (alive on device or bridge to transplantation and recovery) occurred in 82% at 6 months. The patient cohort for implantable continuous (IC) pumps (n = 365) (age 13.2 ± 3.9 years, 18% Interagency Registry for Mechanically Assisted Circulatory Support [Intermacs] profile 1, 23% intubated at implantation, 16% with CHD) was significantly different from the paracorporeal continuous (PC) pump cohort (n = 212) (age 3.6 ± 4.9 years, 46% Intermacs profile 1, 81% intubated, 42% CHD) and the paracorporeal pulsatile (PP) pump cohort (n = 230) (age 2.7 ± 3.5 years, 31% Intermacs profile 1, 76% intubated, 26% CHD). Consistent with their cohort composition, positive outcomes at 6 months based on device type were the following: IC, 92%; PC, 68%; and PP, 81%. The incidence of cerebrovascular accidents in the IC, PC, and PP cohorts was 7%, 14%, and 15%, respectively. IC VADs, the most common VAD type placed in children, are associated with improved outcomes compared with PP and PC devices, though PP and PC devices are limited to supporting our most challenging patients. Noteworthy, the incidence of cerebrovascular accidents for pediatric VADs has significantly decreased and is now 11% overall. This report demonstrates again that although often attributed to age, size, or device type, much of the burden in mortality and adverse events is correlated to the patient's overall state at VAD implantation.


Analysis of Explosives by GC-UV.
A mixture of explosives was analyzed by gas chromatography (GC) linked to ultraviolet (UV) spectrophotometry that enabled detection in the range of 178-330 nm. The gas-phase UV spectra of 2,4,6-trinitrotoluene (TNT), 2,4-dinitrotoluene (DNT), ethylene glycol dinitrate (EGDN), glycerine trinitrate (NG, nitroglycerine), triacetone triperoxide (TATP), and pentaerythritol tetranitrate (PETN) were successfully recorded. The most interesting aspect of the current application is that it enabled simultaneous detection of both the target analyte and its decomposition products. At suitable elevated temperatures of the transfer line between the GC instrument and the UV detector, a partial decomposition was accomplished. Detection was made in real time and resulted in overlaid spectra of the mother compound and its decomposition product. Hence, the presented approach added another level to the qualitative identification of the explosives in comparison with traditional methods that relies only on the detection of the target analyte. As expected, the decomposition product of EGDN, NG, and PETN was NO, while TATP degraded to acetone. DNT and TNT did not exhibit any decomposition at the temperatures used.


MDHM, a macrophage-stimulatory product of Mycoplasma fermentans, leads to in vitro interleukin-1 (IL-1), IL-6, tumor necrosis factor, and prostaglandin production and is pyrogenic in rabbits.
Mycoplasma fermentans-derived high-molecular-weight material (MDHM) was originally discovered because of its capacity to generate, through the induction of monokine synthesis, cytolytic T lymphocytes in concanavalin A-stimulated thymocyte cultures. This study shows that MDHM-activated macrophages not only released interleukin-6 (IL-6) but also exhibited increased synthesis of cell-associated IL-1 as well as liberation of tumor necrosis factor and prostaglandin. We determined 6-keto prostaglandin F1 alpha since it is the stable metabolite of the bioactive prostacyclin. MDHM appeared to be as potent as lipopolysaccharide in inducing the synthesis of these mediators. Priming with gamma interferon further increased MDHM-mediated IL-6 release. Since monokines can be pyrogenic, we tested the effects of an intravenous injection of MDHM on rectal temperatures and leukocyte counts in rabbits. At 1 h after a bolus injection of MDHM, leukocyte counts dropped to about 35% of the initial values, reflecting a decrease in both lymphocytes and granulocytes. At 4 to 6 h after injection, granulocyte counts began to increase again, whereas lymphocyte counts remained low. No leukocytosis was noted during this time. The lack of leukocytosis can be explained by the failure of MDHM-stimulated macrophages to release IL-1. The property of MDHM to cause IL-6 release from macrophages and the IL-6 growth dependency of the 7TD1 hybridoma cell line were made use of in a coculture assay system to quantitate the activity of MDHM. With this method and macrophages from C3H/HeJ lipopolysaccharide-nonresponder mice, MDHM activity was found to be equally distributed in the mycoplasma growth medium and the sedimented mycoplasmas after sonication.


A qualitative evaluation of frontline clinician perspectives toward antibiotic stewardship programs.
To examine the perspectives of caregivers that are not part of the antibiotic stewardship program (ASP) leadership team (eg, physicians, nurses, and clinical pharmacists), but who interact with ASPs in their role as frontline healthcare workers. Qualitative semistructured interviews. The study was conducted in 2 large national healthcare systems including 7 hospitals in the Veterans' Health Administration and 4 hospitals in Intermountain Healthcare. We interviewed 157 participants. The current analysis includes 123 nonsteward clinicians: 47 physicians, 26 pharmacists, 29 nurses, and 21 hospital leaders. Interviewers utilized a semistructured interview guide based on the Consolidated Framework for Implementation Research (CFIR), which was tailored to the participant's role in the hospital as it related to ASPs. Qualitative analysis was conducted using a codebook based on the CFIR. We identified 4 primary perspectives regarding ASPs. (1) Non-ASP pharmacists considered antibiotic stewardship activities to be a high priority despite the added burden to work duties: (2) Nurses acknowledged limited understanding of ASP activities or involvement with these programs; (3) Physicians criticized ASPs for their restrictions on clinical autonomy and questioned the ability of antibiotic stewards to make recommendations without the full clinical picture; And (4) hospital leaders expressed support for ASPs and recognized the unique challenges faced by non-ASP clinical staff. Further understanding these differing perspectives of ASP implementation will inform possible ways to improve ASP implementation across clinical roles.


A 2D metal-organic framework/Ni(OH)2 heterostructure for an enhanced oxygen evolution reaction.
2D metal-organic frameworks (MOFs) are widely regarded as promising electrocatalysts for the oxygen evolution reaction (OER). This results from their inherent properties such as a large portion of surface coordinatively unsaturated metal atoms, rapid mass transfer and enhanced conductivity. However, 2D MOFs have a strong tendency to aggregate, which severely limits their potential application in the OER. Here, novel 2D Ni-BDC/Ni(OH)2 (BDC stands for 1,4-benzenedicarboxylate, C8H4O4) hybrid nanosheets are synthesized via a facile sonication-assisted solution method. Because of the rational material design, the large surface area of Ni-BDC is maintained. Significantly, after coupling, the electronic structure of Ni atoms in the Ni(OH)2 component is well modified, leading to the generation of Ni cations with higher oxidation states, which are desirable for the OER. As-prepared Ni-BDC/Ni(OH)2 exhibits high activity, favorable kinetics and strong durability towards the OER. Specifically, the OER current density of Ni-BDC/Ni(OH)2 is 82.5 mA cm-2 at 1.6 V versus a reversible hydrogen electrode (RHE), which is significantly greater than those of Ni-BDC (5.5 times), Ni(OH)2 (20.6 times) and Ir/C (3.0 times). Moreover, the sonication-assisted method developed in this work can be readily adapted for the preparation of various 2D MOF-based hybrid functional materials.


Latent transformed growth-inhibiting factor in human malignant effusions.
An inhibitor of soft agar colony formation by a human breast carcinoma-derived cell line was found to be present in latent form in the majority of cytology-positive human malignant effusions. Prior to dialysis, addition of human malignant effusions resulted in less than 10% alteration in efficiency of colony formation by the BT-20 human breast carcinoma cell line (mean efficiency 1 colony/4.3 cells plated at 14 days; mean colony diameter greater than 0.8 mm). After dialysis (membrane cutoff of 3500 molecular weight), 58 of 70 malignant effusions from patients with a variety of epithelial cell carcinomas resulted in 71% mean inhibition of colony formation (range 19.1-98% inhibition). Similar inhibition of anchorage-independent growth was observed for a human colon cancer-derived cell line (HCT-15) but not for polyoma and murine sarcoma virus-transformed rodent fibroblast lines. The malignant effusion-related transformed cell growth-inhibiting factor (TGIF) was sensitive to heat, sulfhydryl reduction, and protease treatment. TGIF-containing effusion resulted in parallel inhibition of thymidine incorporation in sensitive cell types in vitro. TGIF was precipitable in 28-34% ammonium sulfate with reconstitution of activity after resolubilization. TGIF was partially purified by chromatography on Biogel A-0.5 and Biogel P-100 which yielded two peaks of inhibitory activity. The predominant species had an approximate molecular weight of 110,000 and could be recovered as a single species from DEAE-cellulose at relatively high salt concentrations (0.4 M NaCl). A smaller amount of inhibitory activity was recovered from Biogel P-100 or Biogel P-60 with an apparent molecular weight of 55,000. The higher molecular-weight TGIF which appears to be a dimer of the Mr 55,000 protein is distinguishable from previously described growth-promoting and -inhibiting factors.


Role of human papillomaviruses in carcinogenesis.
The human papillomavirus (HPV) family comprises more than 170 different types that preferentially infect the mucosa of the genitals, upper-respiratory tract, or the skin. The 'high-risk HPV type', a sub-group of mucosal HPVs, is the cause of approximately 5% of all human cancers, which corresponds to one-third of all virus-induced tumours. Within the high-risk group, HPV16 is the most oncogenic type, being responsible for approximatively 50% of all worldwide cervical cancers. Many studies suggest that, in addition to the high-risk mucosal HPV types, certain cutaneous HPVs also have a role in the development of non-melanoma skin cancer (NMSC). Functional studies on the HPV early gene products showed that E6 and E7 play a key role in carcinogenesis. These two proteins use multiple mechanisms to evade host immune surveillance, allowing viral persistence, and to deregulate cell cycle and apoptosis control, thus facilitating the accumulation of DNA damage and ultimately cellular transformation. The demonstration that high-risk HPV types are the etiological agents of cervical cancer allowed the implementation in the clinical routine of novel screening strategies for cervical lesions, as well as the development of a very efficient prophylactic vaccine. Because of these remarkable achievements, there is no doubt that in the coming decades we will witness a dramatic reduction of cervical cancer incidence worldwide.


Myb-binding protein 1A (MYBBP1A) is essential for early embryonic development, controls cell cycle and mitosis, and acts as a tumor suppressor.
MYBBP1A is a predominantly nucleolar transcriptional regulator involved in rDNA synthesis and p53 activation via acetylation. However little further information is available as to its function. Here we report that MYBBP1A is developmentally essential in the mouse prior to blastocyst formation. In cell culture, down-regulation of MYBBP1A decreases the growth rate of wild type mouse embryonic stem cells, mouse embryo fibroblasts (MEFs) and of human HeLa cells, where it also promotes apoptosis. HeLa cells either arrest at G2/M or undergo delayed and anomalous mitosis. At mitosis, MYBBP1A is localized to a parachromosomal region and gene-expression profiling shows that its down-regulation affects genes controlling chromosomal segregation and cell cycle. However, MYBBP1A down-regulation increases the growth rate of the immortalized NIH3T3 cells. Such Mybbp1a down-regulated NIH3T3 cells are more susceptible to Ras-induced transformation and cause more potent Ras-driven tumors. We conclude that MYBBP1A is an essential gene with novel roles at the pre-mitotic level and potential tumor suppressor activity.


Effects of a delocalizable cation on the headgroup of gemini lipids on the lipoplex-type nanoaggregates directly formed from plasmid DNA.
Lipoplex-type nanoaggregates prepared from pEGFP-C3 plasmid DNA (pDNA) and mixed liposomes, with a gemini cationic lipid (CL) [1,2-bis(hexadecyl imidazolium) alkanes], referred as (C16Im)2Cn (where Cn is the alkane spacer length, n = 2, 3, 5, or 12, between the imidazolium heads) and DOPE zwitterionic lipid, have been analyzed by zeta potential, gel electrophoresis, SAXS, cryo-TEM, fluorescence anisotropy, transfection efficiency, fluorescence confocal microscopy, and cell viability/cytotoxicity experiments to establish a structure-biological activity relationship. The study, carried out at several mixed liposome compositions, α, and effective charge ratios, ρeff, of the lipoplex, demonstrates that the transfection of pDNA using CLs initially requires the determination of the effective charge of both. The electrochemical study confirms that CLs with a delocalizable positive charge in their headgroups yield an effective positive charge that is 90% of their expected nominal one, while pDNA is compacted yielding an effective negative charge which is only 10-25% than that of the linear DNA. SAXS diffractograms show that lipoplexes formed by CLs with shorter spacer (n = 2, 3, or 5) present three lamellar structures, two of them in coexistence, while those formed by CL with longest spacer (n = 12) present two additional inverted hexagonal structures. Cryo-TEM micrographs show nanoaggregates with two multilamellar structures, a cluster-type (at low α value) and a fingerprint-type, that coexist with the cluster-type at moderate α composition. The optimized transfection efficiency (TE) of pDNA, in HEK293T, HeLa, and H1299 cells was higher using lipoplexes containing gemini CLs with shorter spacers at low α value. Each lipid formulation did not show any significant levels of toxicity, the reported lipoplexes being adequate DNA vectors for gene therapy and considerably better than both Lipofectamine 2000 and CLs of the 1,2-bis(hexadecyl ammnoniun) alkane series, recently reported.


The Role of Perspective-Taking on Ability to Recognize Fear.
Impairment in the ability to detect certain emotions, such as fear, is linked to multiple disorders and follows a pattern of inter-individual variability and intra-individual stability over time. Deficits in fear recognition are often related to social and interpersonal difficulties but the mechanisms by which this processing deficit might occur are not well understood. One potential mechanism through which impaired fear detection may influence social competency is through diminished perspective-taking, the ability to perceive and consider the point of view of another individual. In the current study, we hypothesized that intra-individual variability in the accuracy of facial emotion recognition is linked to perspective-taking abilities in a well-characterized, non-clinical adult sample. Results indicated that the ability to accurately detect fear in the faces of others was positively correlated with perspective-taking, consistent with initial hypotheses. This relationship appeared to be unique to recognition of fear, as perspective-taking was not significantly associated with recognition of the other basic emotions. Results from this study represent an initial step towards establishing a potential mechanism between some processes of FER and perspective-taking difficulties. It is important to establish the relationship between these processes in a non-clinical adult sample so that we can consider the possibility of a developmental or pathological influence of impoverished perspective-taking on fear perception.


Facilitators and barriers perceived by health professionals in the implementation of Managing Cancer and Living Meaningfully (CALM) psychotherapy in Santiago.
Palliative care (PC) for advanced cancer is guaranteed by law in Chile, but the formal training for it is insufficient. Training models have emerged internationally that enable professionals to be better prepared for the provision of psychotherapy in PC. The objective of this study is to explore health professionals' perceptions of the 'Managing Cancer and Living Meaningfully' (CALM) psychotherapy and the perceived barriers and facilitators to its implementation, based on a theoretical training. A qualitative study was carried out with health professionals working in oncology and/or PC and participating in a CALM training. A focus group was conducted after to explore the experience of CALM training and the perceived barriers and facilitators to its implementation. A thematic analysis of the content and an analysis of the facilitators and barriers to the implementation of mental health services were carried out. Twenty four professionals participated in the training, six of whom were part of the subsequent focus group. There was a consensus that the training was a positive professional experience and that it is a culturally sensitive and feasible intervention for application in Chile. The barriers identified include institutional bureaucracy as resistance to change, the excess workload of the clinical teams and the absence of spaces for more in-depth training. CALM is a useful and relevant framework for the training of health professionals working in oncology and PC. In Chile, there is a need for training spaces on this topic. Future research and organisational studies should evaluate professionals' beliefs about, and resistance to, adopting evidence-based psychotherapeutic interventions.


Structural evidence for a two-regime photobleaching mechanism in a reversibly switchable fluorescent protein.
Photobleaching, the irreversible photodestruction of a chromophore, severely limits the use of fluorescent proteins (FPs) in optical microscopy. Yet, the mechanisms that govern photobleaching remain poorly understood. In Reversibly Switchable Fluorescent Proteins (RSFPs), a class of FPs that can be repeatedly photoswitched between nonfluorescent and fluorescent states, photobleaching limits the achievable number of switching cycles, a process known as photofatigue. We investigated the photofatigue mechanisms in the protein IrisFP using combined X-ray crystallography, optical in crystallo spectroscopy, mass spectrometry and modeling approaches. At laser-light intensities typical of conventional wide-field fluorescence microscopy, an oxygen-dependent photobleaching pathway was evidenced. Structural modifications induced by singlet-oxygen production within the chromophore pocket revealed the oxidation of two sulfur-containing residues, Met159 and Cys171, locking the chromophore in a nonfluorescent protonated state. At laser-light intensities typical of localization-based nanoscopy (>0.1 kW/cm(2)), a completely different, oxygen-independent photobleaching pathway was found to take place. The conserved Glu212 underwent decarboxylation concomitantly with an extensive rearrangement of the H-bond network around the chromophore, and an sp(2)-to-sp(3) hybridization change of the carbon atom bridging the chromophore cyclic moieties was observed. This two-regime photobleaching mechanism is likely to be a common feature in RSFPs from Anthozoan species, which typically share high structural and sequence identity with IrisFP. In addition, our results suggest that, when such FPs are used, the illumination conditions employed in localization-based super-resolution microscopy might generate less cytotoxicity than those of standard wide-field microscopy at constant absorbed light-dose. Finally, our data will facilitate the rational design of FPs displaying enhanced photoresistance.


Work-related allergic respiratory disease and asthma in spice mill workers is associated with inhalant chili pepper and garlic exposures.
The aim of the study was to determine the prevalence and risk factors for allergic respiratory disease in spice mill workers. A cross-sectional study of 150 workers used European Community Respiratory Health Survey questionnaires, Phadiatop, serum specific IgE (garlic, chili pepper), spirometry and fractional exhaled nitric oxide (FeNO). Personal air samples (n=62) collected from eight-hour shifts were analysed for inhalable particulate mass. Novel immunological assays quantified airborne garlic and chili pepper allergen concentrations. Mean dust particulate mass (geometric mean (GM)=2.06 mg/m(3)), chili pepper (GM=0.44 µg/m(3)) and garlic allergen (GM=0.24 µg/m(3)) were highest in blending and were highly correlated. Workers' mean age was 33 years, 71% were men, 46% current smokers and 45% atopic. Spice-dust-related asthma-like symptoms (17%) were common, as was garlic sensitisation (19%), with 13% being monosensitised and 6% cosensitised to chili pepper. Airflow reversibility and FeNO>50 ppb was present in 4% and 8% of workers respectively. Spice-dust-related ocular-nasal (OR 2.40, CI 1.09 to 5.27) and asthma-like (OR 4.15, CI 1.09 to 15.72) symptoms were strongly associated with airborne garlic in the highly exposed (>0.235 µg/m(3)) workers. Workers monosensitised to garlic were more likely to be exposed to higher airborne chili pepper (>0.92 µg/m(3)) (OR 11.52, CI 1.17 to 113.11) than garlic allergens (OR 5.08, CI 1.17 to 22.08) in this mill. Probable asthma was also more strongly associated with chili pepper than with garlic sensitisation. Exposure to inhalable spice dust (GM >2.06 mg/m(3)) containing garlic (GM>0.24 µg/m³) and chili pepper (GM >0.44 µg/m(3)) allergens increase the risk of allergic respiratory disease and asthma.


Validation of DESS as a DNA Preservation Method for the Detection of Strongyloides spp. in Canine Feces.
Strongyloides stercoralis is a gastrointestinal parasitic nematode with a life cycle that includes free-living and parasitic forms. For both clinical (diagnostic) and environmental evaluation, it is important that we can detect Strongyloides spp. in both human and non-human fecal samples. Real-time PCR is the most feasible method for detecting the parasite in both clinical and environmental samples that have been preserved. However, one of the biggest challenges with PCR detection is DNA degradation during the postage time from rural and remote areas to the laboratory. This study included a laboratory assessment and field validation of DESS (dimethyl sulfoxide, disodium EDTA, and saturated NaCl) preservation of Strongyloides spp. DNA in fecal samples. The laboratory study investigated the capacity of 1:1 and 1:3 sample to DESS ratios to preserve Strongyloides ratti in spike canine feces. It was found that both ratios of DESS significantly prevented DNA degradation compared to the untreated sample. This method was then validated by applying it to the field-collected canine feces and detecting Strongyloides DNA using PCR. A total of 37 canine feces samples were collected and preserved in the 1:3 ratio (sample: DESS) and of these, 17 were positive for Strongyloides spp. The study shows that both 1:1 and 1:3 sample to DESS ratios were able to preserve the Strongyloides spp. DNA in canine feces samples stored at room temperature for up to 56 days. This DESS preservation method presents the most applicable and feasible method for the Strongyloides DNA preservation in field-collected feces.


Ecological consistency across space: a synthesis of the ecological aspects of Dromiciops gliroides in Argentina and Chile.
Dromiciops gliroides is an arboreal marsupial found in the temperate forests of South America (36-43 °S). This species is the sole extant representative of the order Microbiotheria, and is a key seed disperser of many native plant species, including the keystone mistletoe Tristerix corymbosus. Here, we synthesized the current knowledge on the ecological aspects of this species, and compared the available information from Argentina and Chile. Population density (23 ± 2 (mean ± SE) individual/ha) and home range (1.6 ± 0.6 ha) appear to be relatively similar across a marked ecological gradient in the mainland, but lower densities (7 ± 2 individual/ha) and smaller home ranges (0.26 ± 0.04 ha) were detected at island sites. We detected regional variation in body condition in Chile, but there were no significant differences across a wider E-W gradient. Movement patterns fit a random walk model; such behavior might have important consequences in shaping plant's spatial patterns. Although our data suggest that D. gliroides is more tolerant to habitat disturbance than previously thought, its incapability to disperse across non-forested areas suggests that the rapid rate of habitat loss and fragmentation that characterizes southern temperate forests likely poses a serious threat to this species. These ecological similarities are surprising given that forests studied receive dramatically different rainfall and correspond to distinct forest types. The evidence synthetized here dispels some of the myths about this species but also stresses the need for more comprehensive ecological studies across its distribution range.


Thinking glutamatergically: changing concepts of schizophrenia based upon changing neurochemical models.
Clinical concepts of mental illness have always been modulated by underlying theoretical considerations. For the past fifty years, schizophrenia has been considered primarily a disease of dopaminergic neurotransmission. Although this conceptualization has helped greatly in explaining the clinical effects of psychostimulants and guiding the clinical use of both typical and atypical antipsychotics, it has nevertheless shaded how we look at the disorder from both a pathophysiological and therapeutic perspective. For example, most explanatory research in schizophrenia has focused on dopamine-rich regions of the brain, with little investigation of regions of the brain that are relatively dopamine poor. Starting approximately twenty years ago, an alternative formulation of schizophrenia was proposed based upon actions of the "dissociative anesthetic" class of psychotomimetic agents, including phencyclidine (PCP), ketamine and various designer drugs. These compounds induce psychosis by blocking neurotransmission at N-methyl-D-aspartate (NMDA)-type glutamate receptors, suggesting an alternative model for pathogenesis in schizophrenia. As opposed to dopamine, the glutamatergic system is widely distributed throughout the brain and plays a prominent role in sensory processing as well as in subsequent stages of cortical analysis. Glutamatergic theories of schizophrenia, thus, predict that cortical dysfunction will be regionally diffuse but process specific. In addition, NMDA receptors incorporate binding sites for specific endogenous brain compounds, including the amino acids glycine and D-serine and the redox modulator glutathione, and interact closely with dopaminergic, cholinergic and γ-aminobutyric acid (GABA)-ergic systems. Glutamatergic theories, thus, open new potential approaches for treatment of schizophrenia, most of which are only now entering clinical evaluation.


Morphodynamics of phyllotaxis.
A morphodynamic model for phyllotaxis based upon an axiomatic approach is presented. We show that the helical forms of alternate phyllotaxis can be derived from the assumption of the rudiment's growth and movement on the cylindrical embryo surface in the absence of a longitudinal displacement. This leads to the repeating transition of tetragonal packaging of the rudiments into hexagonal packaging and vice versa. Under these conditions, sequences of rudiments produce either left-handed or right-handed helices, the number of which at the circumference of the cylinder corresponds to adjacent numbers of the Fibonacci series. Cross-opposite phyllotaxis forms are defined as superior with respect to the alternate ones, and verticillate phyllotaxis forms as superior with respect to the cross-opposite ones. Different phyllotaxis forms can be interpreted as a result of stretching of crystalline structures of the embryo formed by dense packing of rudiments. The superior phyllotaxis forms can be considered as the additive summation of lower forms. Morphodynamic mechanisms underlying the formation of multiple forms of helical phyllotaxis are discussed.


Total hip replacement.
To provide physicians with a current consensus on total hip replacement. A non-Federal, nonadvocate, 13-member consensus panel representing the fields of orthopedic surgery, rehabilitation and physical medicine, biomechanics and biomaterials, internal medicine, public health, geriatrics and biostatistics, and a public representative. In addition, 27 experts in orthopedic surgery, rehabilitation and physical medicine, biomechanics and biomaterials, rheumatology, geriatrics, and epidemiology presented data to the consensus panel and a conference audience of 425. The literature was search through Medline and an extensive bibliography of references was provided to the panel and the conference audience. Experts prepared abstracts with relevant citations from the literature. Scientific evidence was given precedence over clinical anecdotal experience. The panel, answering predefined consensus questions, developed their conclusions based on the scientific evidence presented in open forum and the scientific literature. The panel composed a draft statement that was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. Total hip replacement is an option for nearly all patients with diseases of the hip that cause chronic discomfort and significant functional impairment. Most patients have an excellent prognosis for long-term improvement in symptoms and physical function. At this time, a cemented femoral component using modern cementing techniques, paired with a porous-coated acetabular component, can give excellent long-term results. Revision of a total hip replacement is indicated when mechanical failure occurs. Continued periodic follow-up is necessary to identify early evidence of impending failure so as to permit remedial action before a catastrophic event.


A retrospective study of the long-term survival of RESTORE® dental implants with resorbable blast media surface.
The aim of this study was to retrospectively evaluate the survival and failure rates of RESTORE® implants over a follow-up period of 10-15 years at a university dental hospital and to investigate the factors affecting the survival rate of these dental implants. A total of 247 RESTORE® dental implants with a resorbable blast media (RBM) surface inserted in 86 patients between March 2006 and April 2011 at the Department of Periodontology of Seoul National University Dental Hospital were included. Patients with follow-up periods of less than 10 years were excluded, and data analysis was conducted based on dental records and radiographs. Over a 10- to 15-year period, the cumulative survival rate of the implants was 92.5%. Seventeen implants (6.88%) were explanted due to implant fracture (n=10, 4.05%), peri-implantitis (n=6, 2.43%), and screw fracture (n=1, 0.4%). The results of univariate regression analysis using a Cox proportional hazards model demonstrated that implants placed in male patients (hazard ratio [HR], 4.542; 95% confidence interval [CI], 1.305-15.807; P=0.017) and implants that supported removable prostheses (HR, 15.498; 95% CI, 3.105-77.357; P=0.001) showed statistically significant associations with implant failure. Within the limitations of this retrospective study, the RESTORE® dental implant with an RBM surface has a favorable survival rate with stable clinical outcomes.


RAF kinase inhibitory protein (RKIP) modulates cell cycle kinetics and motility.
RKIP-1 is a metastasis suppressor that is frequently downregulated in aggressive cancers. However, the consequences of RKIP loss in primary or immortalized cells have not yet been explored. Using HEK-293 RKIP depleted (termed HEK-499) and Flp-In T-Rex-293 RKIP inducible cell lines combined with whole transcriptome analysis, we show that RKIP-1 silencing accelerates DNA synthesis and G1/S transition entry by inducing the expression of cdc6, MCM 2, 4, 6, 7, cdc45L, cyclin D2, cyclin E2, cyclin D1, SKP2 and the downregulation of p21(cip1). Moreover, RKIP depletion accelerates the time from nuclear envelop breakdown (NEB) to anaphase markedly, while the upregulation of RKIP shortened the NEB to anaphase time. We show that RKIP depletion induces the expression of NEK6, a molecule known to enhance G2/M transition, and down-regulates G2/M checkpoint molecules like Aurora B, cyclin G1 and sertuin that slow the G2/M transition time. These subtle changes in the kinetics of the cell cycle culminate in a higher proliferation rate of HEK-499 compared to control cells. Finally, we show that RKIP depletion enhances cellular motility by inducing the expression/stabilization of β-catenin, vimentin, MET and PAK1. Overall, our data suggest that modulation of the cell cycle checkpoints and motility by RKIP may be fundamental to its metastasis suppressive function in cancer and that RKIP role in a cell is more intricate and diverse than previously thought.


Cellular Bioenergetics and AMPK and TORC1 Signalling in Blood Lymphoblasts Are Biomarkers of Clinical Status in FMR1 Premutation Carriers.
Fragile X Associated Tremor/Ataxia Syndrome (FXTAS) is a neurodegenerative disorder affecting carriers of premutation alleles (PM) of the X-linked FMR1 gene, which contain CGG repeat expansions of 55-200 range in a non-coding region. This late-onset disorder is characterised by the presence of tremor/ataxia and cognitive decline, associated with the white matter lesions throughout the brain, especially involving the middle cerebellar peduncles. Nearly half of older male and ~ 20% of female PM carriers develop FXTAS. While there is evidence for mitochondrial dysfunction in neural and some peripheral tissues from FXTAS patients (though less obvious in the non-FXTAS PM carriers), the results from peripheral blood mononuclear cells (PBMC) are still controversial. Motor, cognitive, and neuropsychiatric impairments were correlated with measures of mitochondrial and non-mitochondrial respiratory activity, AMPK, and TORC1 cellular stress-sensing protein kinases, and CGG repeat size, in a sample of adult FXTAS male and female carriers. Moreover, the levels of these cellular measures, all derived from Epstein- Barr virus (EBV)- transformed and easily accessible blood lymphoblasts, were compared between the FXTAS (N = 23) and non-FXTAS (n = 30) subgroups, and with baseline data from 33 healthy non-carriers. A significant hyperactivity of cellular bioenergetics components as compared with the baseline data, more marked in the non-FXTAS PMs, was negatively correlated with repeat numbers at the lower end of the CGG-PM distribution. Significant associations of these components with motor impairment measures, including tremor-ataxia and parkinsonism, and neuropsychiatric changes, were prevalent in the FXTAS subgroup. Moreover, a striking elevation of AMPK activity, and a decrease in TORC1 levels, especially in the non-FXTAS carriers, were related to the size of CGG expansion. The bioenergetics changes in blood lymphoblasts are biomarkers of the clinical status of FMR1 carriers. The relationship between these changes and neurological involvement in the affected carriers suggests that brain bioenergetic alterations are reflected in this peripheral tissue. A possible neuroprotective role of stress sensing kinase, AMPK, in PM carriers, should be addressed in future longitudinal studies. A decreased level of TORC1-the mechanistic target of the rapamycin complex, suggests a possible future approach to therapy in FXTAS.


Transfusional iron overload in children with sickle cell anemia on chronic transfusion therapy for secondary stroke prevention.
Chronic transfusion reduces the risk of recurrent stroke in children with sickle cell anemia (SCA) but leads to iron loading. Management of transfusional iron overload in SCA has been reported as suboptimal [1], but studies characterizing monitoring and treatment practices for iron overload in children with SCA, particularly in recent years with the expansion of chelator options, are lacking. We investigated the degree of iron loading and treatment practices of 161 children with SCA receiving transfusions for a history of stroke who participated in the Stroke with Transfusions Changing to Hydroxyurea (SWiTCH) trial. Data obtained during screening, including past and entry liver iron concentration (LIC) measurements, ferritin values, and chelation were analyzed. The mean age at enrollment was 12.9 ± 4 years and the mean duration of transfusion was 7 ± 3.8 years. Baseline LIC (median 12.94 mg/g dw) and serum ferritin (median 3,164 ng/mL) were elevated. Chelation therapy was initiated after a mean of 2.6 years of transfusions. At study entry, 137 were receiving chelation, most of whom (90%) were receiving deferasirox. This study underscores the need for better monitoring of iron burden with timely treatment adjustments in chronically transfused children with SCA.


Estimating mineral requirements of Nellore beef bulls fed with or without inorganic mineral supplementation and the influence on mineral balance.
The objectives of this study were to quantify the mineral balance of Nellore cattle fed with and without Ca, P, and micromineral (MM) supplementation and to estimate the net and dietary mineral requirement for cattle. Nellore cattle ( = 51; 270.4 ± 36.6 kg initial BW and 8 mo age) were assigned to 1 of 3 groups: reference ( = 5), maintenance ( = 4), and performance ( = 42). The reference group was slaughtered prior to the experiment to estimate initial body composition. The maintenance group was used to collect values of animals at low gain and reduced mineral intake. The performance group was assigned to 1 of 6 treatments: sugarcane as the roughage source with a concentrate supplement composed of soybean meal and soybean hulls with and without Ca, P, and MM supplementation; sugarcane as the roughage source with a concentrate supplement composed of soybean meal and ground corn with and without Ca, P, and MM supplementation; and corn silage as the roughage source with a concentrate supplement composed of soybean meal and ground corn with and without Ca, P, and MM supplementation. Orthogonal contrasts were adopted to compare mineral intake, fecal and urinary excretion, and apparent retention among treatments. Maintenance requirements and true retention coefficients were generated with the aid of linear regression between mineral intake and mineral retention. Mineral composition of the body and gain requirements was assessed using nonlinear regression between body mineral content and mineral intake. Mineral intake and fecal and urinary excretion were measured. Intakes of Ca, P, S, Cu, Zn, Mn, Co, and Fe were reduced in the absence of Ca, P, and MM supplementation ( < 0.05). Fecal excretion of Ca, Cu, Zn, Mn, and Co was also reduced in treatments without supplementation ( < 0.01). Overall, excretion and apparent absorption and retention coefficients were reduced when minerals were not supplied ( < 0.05). The use of the true retention coefficient instead of the true absorption coefficient provided a better estimate of mineral requirements. Dietary mineral requirements were lower for P, Cu, and Zn and greater for Fe compared with previously published recommendations. This study provides useful information about mineral requirements and mineral supplementation to obtain adequate dietary mineral supply of Nellore cattle in tropical conditions.


Conservation of coding potential and terminal sequences in four different isolates of Borna disease virus.
We determined the complete nucleotide sequences of two poorly characterized strains of Borna disease virus (BDV) and compared them to reference strains V and He/80. Strain H1766 was almost 98% and 95% identical to strains V and He/80, respectively, whereas strain No/98 was only about 81% identical to both reference strains. In contrast to earlier reports, we found an additional A residue at the extreme 3'-end of the single-stranded RNA genome in all four BDV strains. The exact numbers of nucleotides in the four BDV genomes could not be determined due to a micro-heterogeneity at the 5'-end. If our longest sequence is a correct copy of the viral RNA, the two ends of the BDV genome would show almost perfect complementarity. All three transcription start sites, all four termination sites, both splice donor sites and both major splice acceptor sites are highly conserved, whereas a minor alternative splice acceptor site is not. The L protein of No/98 differs at 7% of its amino acid positions from the polymerase in the other strains, with most differences mapping to the C-terminal moiety of the molecule. Re-evaluation of L protein sequences of strains V and He/80 revealed differences at several positions compared to published information, indicating that variant forms of the viral polymerase have previously been characterized. These results are important because correct structures of genome ends and of the polymerase gene are the most critical parameters for the future development of techniques that will permit the genetic manipulation of BDV.


A web-based intervention for abused women: the New Zealand isafe randomised controlled trial protocol.
Intimate partner violence (IPV) and its associated negative mental health consequences are significant for women in New Zealand and internationally. One of the most widely recommended interventions is safety planning. However, few women experiencing violence access specialist services for safety planning. A safety decision aid, weighing the dangers of leaving or staying in an abusive relationship, gives women the opportunity to prioritise, plan and take action to increase safety for themselves and their children. This randomised controlled trial is testing the effectiveness of an innovative, interactive web-based safety decision aid. The trial is an international collaborative concurrent replication of a USA trial (IRIS study NCT01312103), regionalised for the Aotearoa New Zealand culture and offers fully automated online trial recruitment, eligibility screening and consent. In a fully automated web-based trial (isafe) 340 abused women will be randomly assigned in equal numbers to a safety decision aid intervention or usual safety planning control website. Intervention components include: (a) safety priority setting, (b) danger assessment and (c) an individually tailored safety action plan. Self-reported outcome measures are collected at baseline and 3, 6, and 12-months post-baseline. Primary outcomes are depression (measured by Center for Epidemiologic Studies Depression Scale, Revised) and IPV exposure (measured by Severity Violence Against Women Scale) at 12 months post-baseline. Secondary outcomes include PTSD, psychological abuse, decisional conflict, safety behaviors and danger in the relationship. This trial will provide much-needed information on the potential relationships among safety planning, improved mental health, reduced violence as well as decreased decisional conflict related to safety in the abusive relationship. The novel web-based safety decision aid intervention may provide a cost-effective, easily accessed safety-planning resource that can be translated into clinical and community practice by multiple health disciplines and advocates. The trial will also provide information about how women in abusive relationships safely access safety information and resources through the Internet. Finally, the trial will inform other research teams on the feasibility and acceptability of fully automated recruitment, eligibility screening, consent and retention procedures. Trial registered on 03 July 2012 on the Australian New Zealand Clinical Trials Registry ACTRN12612000708853 .


Determinants of choosing public or private health care among patients with sexually transmitted infections in Uganda.
To identify variables that distinguish patients with sexually transmitted infections (STIs) who seek care in public or private health units. To recommend measures for improved care of patients with STIs. Patients with STIs were interviewed at public (n = 101) or private health units (n = 124). Information was collected on attitudinal, normative, and self-efficacy beliefs; STI symptoms; health-seeking behavior; sociodemographic characteristics, and on partner referral. Choosing private health units is favored by age >25 years, favorable beliefs towards private health units (e.g., they cure or prevent STIs and give adequate drug doses); unfavorable beliefs towards public health units (such as they make STIs chronic and have corrupt staff); not being influenced by sexual partner(s) in choice of treatment site, being likely to chose a treatment site if sexual partners were not treated free, and being likely to choose a treatment site if not recommend by a friend. This 9-variable model correctly classified 214 (95%) of the 225 patients (model chi squared = 192, 9 degrees of freedom, P <0.001). Psychosocial variables markedly influence choice of health care provider. Improving quality of care will enhance STI management and help to modify the unfavorable psychosocial beliefs.


Allele frequencies and minor contributor match statistic convergence using simulated population replicates.
Probabilistic genotyping permits a comparison of forensic evidence given hypotheses regarding the origin of observed short tandem repeat alleles in a mixed DNA profile. Using the publicly available R package forensim, it has been proposed that mixtures with non-contributors from low genetic diversity populations are more likely to be mistakenly identified as contributors to a mixture than non-contributors from high genetic diversity populations. We hypothesized that these observations are attributed to the unique distribution of alleles in the reference population and may not generalize to other samplings of the same population. We used forensim to simulate 200 US populations (50 each of self-reported African-American, Asian-American, European-American, and Hispanic descent). We compared likelihood ratios for 2400 mixtures to those derived from published data and identified stark differences. A minimum of ten population replicates were required to reduce observed differences relative to published data. Deviations from Hardy-Weinberg equilibrium and allele frequency distributions suggest that simulated populations should be sufficiently evaluated for expectations of population genetic parameters prior to use in DNA mixture modeling experiments. Overall, our findings support the utility of forensim and further describe its suitability to model population genetic parameters but suggest that a single population replicate (directly ascertained or simulated) may be insufficient to make conclusions about a given DNA mixture.


Time-resolved microfocused small-angle X-ray scattering investigation of the microfluidic concentration of charged nanoparticles.
We describe the concentration process of a dispersion of silica nanoparticles undergoing evaporation in a dedicated microfluidic device. Using microfocused small-angle X-ray scattering, we measure in time and space both the concentration field of the dispersion and its structure factor. We show that the electrostatic interactions affect the concentration rate by strongly enhancing the collective diffusion coefficient of the nanoparticle dispersion. En route towards high concentrations, the nanoparticles eventually undergo a liquid-solid phase transition in which we evidence crystallites of micron size.


The Use of Social Media to Increase the Impact of Health Research: Systematic Review.
Academics in all disciplines increasingly use social media to share their publications on the internet, reaching out to different audiences. In the last few years, specific indicators of social media impact have been developed (eg, Altmetrics), to complement traditional bibliometric indicators (eg, citation count and h-index). In health research, it is unclear whether social media impact also translates into research impact. The primary aim of this study was to systematically review the literature on the impact of using social media on the dissemination of health research. The secondary aim was to assess the correlation between Altmetrics and traditional citation-based metrics. We conducted a systematic review to identify studies that evaluated the use of social media to disseminate research published in health-related journals. We specifically looked at studies that described experimental or correlational studies linking the use of social media with outcomes related to bibliometrics. We searched the Medical Literature Analysis and Retrieval System Online (MEDLINE), Excerpta Medica dataBASE (EMBASE), and Cumulative Index to Nursing and Allied Health Literature (CINAHL) databases using a predefined search strategy (International Prospective Register of Systematic Reviews: CRD42017057709). We conducted independent and duplicate study selection and data extraction. Given the heterogeneity of the included studies, we summarized the findings through a narrative synthesis. Of a total of 18,624 retrieved citations, we included 51 studies: 7 (14%) impact studies (answering the primary aim) and 44 (86%) correlational studies (answering the secondary aim). Impact studies reported mixed results with several limitations, including the use of interventions of inappropriately low intensity and short duration. The majority of correlational studies suggested a positive association between traditional bibliometrics and social media metrics (eg, number of mentions) in health research. We have identified suggestive yet inconclusive evidence on the impact of using social media to increase the number of citations in health research. Further studies with better design are needed to assess the causal link between social media impact and bibliometrics.


Tocotrienol is the most effective vitamin E for reducing endothelial expression of adhesion molecules and adhesion to monocytes.
Alpha-tocopherol and its esterified derivatives have been shown to be effective in reducing monocytic-endothelial cell adhesion. However, the effect of alpha-tocotrienol (alpha-T3) has not been characterized. In the present study, using human umbilical vein endothelial cells (HUVEC) as the model system, we examined the relative inhibitory effects of alpha-T3 and other vitamin E derivatives on cell surface adhesion molecule expression under TNF-alpha stimulation. Using enzyme-linked immunosorbent assay, we demonstrated that alpha-T3 markedly inhibited the surface expression of vascular cell adhesion molecule-1 in TNF-alpha activated HUVEC in a dose- and time-dependent manner. The optimal inhibition was observed at 25 micromol/l alpha-T3 within 24 h (77+/-5%) without cytotoxicity. In addition, the surface expression of intercellular adhesion molecule-1 and E-selectin were also reduced by 40+/-7 and 42+/-5%, respectively. In order to further evaluate the effects of alpha-T3 on the vascular endothelium, we investigated the ability of monocytes to adhere to endothelial cells. Interestingly, a 63+/-3% decrease in monocytic cell adherence was observed. Compared to alpha-tocopherol and alpha-tocopheryl succinate, alpha-T3 displayed a more profound inhibitory effect on adhesion molecule expression and monocytic cell adherence. This inhibitory action by alpha-T3 on TNF-alpha-induced monocyte adhesion was shown to be NF-kappaB dependent and was interestingly reversed with co-incubation with farnesol and geranylgeraniol, suggesting a role for prenylated proteins in the regulation of adhesion molecule expression. In summary, the above results suggest that alpha-T3 is a potent and effective agent in the reduction of cellular adhesion molecule expression and monocytic cell adherence.


Methods for increasing the rate of anammox attachment in a sidestream deammonification MBBR.
Deammonification (partial nitritation-anammox) is a proven process for the treatment of high-nitrogen waste streams, but long startup time is a known drawback of this technology. In a deammonification moving bed biofilm reactor (MBBR), startup time could potentially be decreased by increasing the attachment rate of anammox bacteria (AMX) on virgin plastic media. Previous studies have shown that bacterial adhesion rates can be increased by surface modification or by the development of a preliminary biofilm. This is the first study on increasing AMX attachment rates in a deammonification MBBR using these methods. Experimental media consisted of three different wet-chemical surface treatments, and also media transferred from a full-scale mainstream fully nitrifying integrated fixed-film activated sludge (IFAS) reactor. Following startup of a full-scale deammonification reactor, the experimental media were placed in the full-scale reactor and removed for activity rate measurements and biomass testing after 1 and 2 months. The media transferred from the IFAS process exhibited a rapid increase in AMX activity rates (1.1 g/m(2)/day NH(4)(+) removal and 1.4 g/m(2)/day NO(2)(-) removal) as compared to the control (0.2 g/m(2)/day NH(4)(+) removal and 0.1 g/m(2)/day NO(2)(-) removal) after 1 month. Two out of three of the surface modifications resulted in significantly higher AMX activity than the control at 1 and 2 months. No nitrite oxidizing bacteria activity was detected in either the surface modified media or IFAS media batch tests. The results indicate that startup time of a deammonification MBBR could potentially be decreased through surface modification of the plastic media or through the transfer of media from a mature IFAS process.


Organohalogenated contaminants in white-tailed eagle (Haliaeetus albicilla) nestlings: An assessment of relationships to immunoglobulin levels, telomeres and oxidative stress.
Biomagnifying organohalogenated compounds (OHCs) may have adverse effects on the health of birds, especially marine avian top predators that accumulate high OHC loads. Contaminants may impair the humoral immunity and also influence the antioxidant enzyme activity (i.e. oxidative stress). Moreover, physical conditions and oxidative stress during development may reduce telomere lengths, one of the main mechanisms explaining cell senescence. To examine the potential effects of environmental contaminants on physiological biomarkers of health, OHCs with different 'physicochemical' properties were related to immunoglobulin Y levels (IgY; humoral immunity), superoxide dismutase enzyme (SOD) activity in blood plasma, and telomere length (measured in red blood cells) in individual 7-8weeks old nestlings (n=35) of white-tailed eagles (Haliaeetus albicilla) in the Norwegian Sub-Arctic. Different organochlorines (OCs) and perfluoroalkylated substances (PFASs) were measured in blood plasma of nestlings, demonstrating higher concentrations of the emerging contaminants (PFASs), notably perfluorooctane sulfonate (PFOS), compared to legacy OCs. There were no relationships between the contaminant loads and plasma IgY levels. Moreover, differences between years were found for telomere lengths, but this was not related to contaminants and more likely a result of different developmental conditions. However, there were significant and negative relationships between the OC loadings and the SOD activity. This suggests that some legacy OCs challenge the antioxidant capacity in nestlings of white-tailed eagles.


Twitter Analytics to Inform Provisional Guidance for COVID-19 Challenges in the Meatpacking Industry.
The COVID-19 pandemic raised considerable challenges to obtain reliable guidance to help occupational health practitioners, workers, and stakeholders building up efficient prevention strategies at the workplace, between the constant increase of publications in the domain, the time required to run high-quality research and systematic reviews, and the urgent need to identify areas for prevention at the workplace. Social Media and Twitter, in particular, have already been used in research and constitute a useful source of information to identify community needs and topics of interest for prevention in the meatpacking industry. In this commentary, we introduce the methods and tools we used to screen relevant posts on Twitter. Twitter analytics is a way to capture real-time concerns of the community and help ensure compliance with the notion of social accountability. As such research has limitations in terms of exhaustiveness and level of evidence, it should be considered as provisional guidance to direct both actions at the workplace and further conventional research projects.


Extracellular cyclophilin A possesses chemotaxic activity in cattle.
Cyclophilin A (CyPA) was originally discovered in bovine thymocytes as a cytosolic binding protein of the immunosuppressive drug cyclosporine A. Recent studies have revealed that in mice and humans, CyPA is secreted from cells in injured or infected tissues and plays a role in recruiting inflammatory cells in those tissues. Here we found that in cattle abundant level of extracellular CyPA was observed in tissues with inflammation. To aid in investigating the role of extracellular CyPA in cattle, we generated recombinant bovine CyPA (rbCyPA) and tested its biological activity as an inflammatory mediator. When bovine peripheral blood cells were treated with rbCyPA in vitro, we observed that rbCyPA reacts with the membranous surface of granulocytes, monocytes and lymphocytes. Chemotaxis analysis showed that the granulocytes migrate toward rbCyPA and the migration is inhibited by pre-treatment with an anti-bovine CyPA antibody. These results indicate that, as for mice and humans, extracellular CyPA possesses chemotactic activity to recruit inflammatory cells (e.g., granulocytes) in cattle, and could thus be a potential therapeutic target for the treatment of inflammation.


Aminoglycoside-modifying enzymes determine the innate susceptibility to aminoglycoside antibiotics in rapidly growing mycobacteria.
Infections caused by the rapidly growing mycobacterium (RGM) Mycobacterium abscessus are notoriously difficult to treat due to the innate resistance of M. abscessus to most clinically available antimicrobials. Aminoglycoside antibiotics (AGA) are a cornerstone of antimicrobial chemotherapy against M. abscessus infections, although little is known about intrinsic drug resistance mechanisms. We investigated the role of chromosomally encoded putative aminoglycoside-modifying enzymes (AME) in AGA susceptibility in M. abscessus. Clinical isolates of M. abscessus were tested for susceptibility to a series of AGA with different substituents at positions 2', 3' and 4' of ring 1 in MIC assays. Cell-free extracts of M. abscessus type strain ATCC 19977 and Mycobacterium smegmatis strains SZ380 [aac(2')-Id(+)], EP10 [aac(2')-Id(-)] and SZ461 [aac(2')-Id(+), rrs A1408G] were investigated for AGA acetylation activity using thin-layer chromatography (TLC). Cell-free ribosome translation assays were performed to directly study drug-target interaction. Cell-free translation assays demonstrated that ribosomes of M. abscessus and M. smegmatis show comparable susceptibility to all tested AGA. MIC assays for M. abscessus and M. smegmatis, however, consistently showed the lowest MIC values for 2'-hydroxy-AGA as compared with 2'-amino-AGA, indicating that an aminoglycoside-2'-acetyltransferase, Aac(2'), contributes to innate AGA susceptibility. TLC experiments confirmed enzymatic activity consistent with Aac(2'). Using M. smegmatis as a model for RGM, acetyltransferase activity was shown to be up-regulated in response to AGA-induced inhibition of protein synthesis. Our findings point to AME as important determinants of AGA susceptibility in M. abscessus.


Shared decision-making following disclosure of coronary heart disease genetic risk: results from a randomized clinical trial.
Whether disclosure of genetic risk for coronary heart disease (CHD) influences shared decision-making (SDM) regarding use of statins to reduce CHD risk is unknown. We randomized 207 patients, age 45-65 years, at intermediate CHD risk, and not on statins, to receive the 10-year risk of CHD based on conventional risk factors alone (n=103) or in combination with a genetic risk score (n=104). A genetic counselor disclosed this information followed by a physician visit for SDM regarding statin therapy. A novel decision aid was used in both encounters to disclose the CHD risk estimates and facilitate SDM regarding statin use. Patients reported their decision quality and physician visit satisfaction using validated surveys. There were no statistically significant differences between the two groups in the SDM score, satisfaction with the clinical encounter, perception of the quality of the discussion or of participation in decision-making and physician visit satisfaction scores. Quantitative analyses of a random subset of 80 video-recorded encounters using the OPTION5 scale also showed no significant difference in SDM between the two groups. Disclosure of CHD genetic risk using an electronic health record-linked decision aid did not adversely affect SDM or patients' satisfaction with the clinical encounter. NCT01936675; Results.


Enlarging the toolbox for allergen epitope definition with an allergen-type model protein.
Birch pollen-allergic subjects produce polyclonal cross-reactive IgE antibodies that mediate pollen-associated food allergies. The major allergen Bet v 1 and its homologs in plant foods bind IgE in their native protein conformation. Information on location, number and clinical relevance of IgE epitopes is limited. We addressed the use of an allergen-related protein model to identify amino acids critical for IgE binding of PR-10 allergens. Norcoclaurine synthase (NCS) from meadow rue is structurally homologous to Bet v 1 but does not bind Bet v 1-reactive IgE. NCS was used as the template for epitope grafting. NCS variants were tested with sera from 70 birch pollen allergic subjects and with monoclonal antibody BV16 reported to compete with IgE binding to Bet v 1. We generated an NCS variant (Δ29NCSN57/I58E/D60N/V63P/D68K) harboring an IgE epitope of Bet v 1. Bet v 1-type protein folding of the NCS variant was evaluated by 1H-15N-HSQC NMR spectroscopy. BV16 bound the NCS variant and 71% (50/70 sera) of our study population showed significant IgE binding. We observed IgE and BV16 cross-reactivity to the epitope presented by the NCS variant in a subgroup of Bet v 1-related allergens. Moreover BV16 blocked IgE binding to the NCS variant. Antibody cross-reactivity depended on a defined orientation of amino acids within the Bet v 1-type conformation. Our system allows the evaluation of patient-specific epitope profiles and will facilitate both the identification of clinically relevant epitopes as biomarkers and the monitoring of therapeutic outcomes to improve diagnosis, prognosis, and therapy of allergies caused by PR-10 proteins.


Genome-wide footprints of pig domestication and selection revealed through massive parallel sequencing of pooled DNA.
Artificial selection has caused rapid evolution in domesticated species. The identification of selection footprints across domesticated genomes can contribute to uncover the genetic basis of phenotypic diversity. Genome wide footprints of pig domestication and selection were identified using massive parallel sequencing of pooled reduced representation libraries (RRL) representing ∼2% of the genome from wild boar and four domestic pig breeds (Large White, Landrace, Duroc and Pietrain) which have been under strong selection for muscle development, growth, behavior and coat color. Using specifically developed statistical methods that account for DNA pooling, low mean sequencing depth, and sequencing errors, we provide genome-wide estimates of nucleotide diversity and genetic differentiation in pig. Widespread signals suggestive of positive and balancing selection were found and the strongest signals were observed in Pietrain, one of the breeds most intensively selected for muscle development. Most signals were population-specific but affected genomic regions which harbored genes for common biological categories including coat color, brain development, muscle development, growth, metabolism, olfaction and immunity. Genetic differentiation in regions harboring genes related to muscle development and growth was higher between breeds than between a given breed and the wild boar. These results, suggest that although domesticated breeds have experienced similar selective pressures, selection has acted upon different genes. This might reflect the multiple domestication events of European breeds or could be the result of subsequent introgression of Asian alleles. Overall, it was estimated that approximately 7% of the porcine genome has been affected by selection events. This study illustrates that the massive parallel sequencing of genomic pools is a cost-effective approach to identify footprints of selection.


Cable Tension Optimization for an Epicardial Parallel Wire Robot.
HeartPrinter is a novel under-constrained 3-cable parallel wire robot designed for minimally invasive epicardial interventions. The robot adheres to the beating heart using vacuum suction at its anchor points, with a central injector head that operates within the triangular workspace formed by the anchors, and is actuated by cables for multipoint direct gene therapy injections. Minimizing cable tensions can reduce forces on the heart at the anchor points while supporting rapid delivery of accurate injections and minimizing procedure time, risk of damage to the robot, and strain to the heart. However, cable tensions must be sufficient to hold the injector head's position as the heart moves and to prevent excessive cable slack. We pose a linear optimization problem to minimize the sum of cable tension magnitudes for HeartPrinter while ensuring the injector head is held in static equilibrium and the tensions are constrained within a feasible range. We use Karush-Kuhn-Tucker optimality conditions to derive conditional algebraic expressions for optimal cable tensions as a function of injector head position and workspace geometry, and we identify regions of injector head positions where particular combinations of cable tensions are optimally at minimum allowable tensions. The approach can rapidly solve for the minimum set of cable tensions for any robot workspace geometry and injector head position and determine whether an injection site is attainable.


Evaluation of Kerr constant of blue-phase liquid crystals by measuring off-axis retardation in vertical electric field cells.
Because of the nonuniform electric field of the in-plane-switching cell in the thickness direction, an accurate and efficient way for evaluating the Kerr constant of blue-phase liquid crystal (BPLC) needs to be developed. This study demonstrates a method for evaluating the Kerr constant by measuring the off-axis-induced retardation (R(th)) change in normal vertical field cells using a commercial polarimeter. The angle-dependent behavior of the R(th) change is observed as an electric-tunable positive C retarder. In this paper, a sigmoid fitting model has been chosen for calculating the Kerr constant for considering the very small intrinsic birefringence of the BPLC.