Microarray and cDNA sequence analysis of transcription during nerve-dependent limb regeneration.
Microarray analysis and 454 cDNA sequencing were used to investigate a centuries-old problem in regenerative biology: the basis of nerve-dependent limb regeneration in salamanders. Innervated (NR) and denervated (DL) forelimbs of Mexican axolotls were amputated and transcripts were sampled after 0, 5, and 14 days of regeneration. Considerable similarity was observed between NR and DL transcriptional programs at 5 and 14 days post amputation (dpa). Genes with extracellular functions that are critical to wound healing were upregulated while muscle-specific genes were downregulated. Thus, many processes that are regulated during early limb regeneration do not depend upon nerve-derived factors. The majority of the transcriptional differences between NR and DL limbs were correlated with blastema formation; cell numbers increased in NR limbs after 5 dpa and this yielded distinct transcriptional signatures of cell proliferation in NR limbs at 14 dpa. These transcriptional signatures were not observed in DL limbs. Instead, gene expression changes within DL limbs suggest more diverse and protracted wound-healing responses. 454 cDNA sequencing complemented the microarray analysis by providing deeper sampling of transcriptional programs and associated biological processes. Assembly of new 454 cDNA sequences with existing expressed sequence tag (EST) contigs from the Ambystoma EST database more than doubled (3935 to 9411) the number of non-redundant human-A. mexicanum orthologous sequences. Many new candidate gene sequences were discovered for the first time and these will greatly enable future studies of wound healing, epigenetics, genome stability, and nerve-dependent blastema formation and outgrowth using the axolotl model.


Depression during pegylated interferon-alpha plus ribavirin therapy: prevalence and prediction.
Interferon-alpha (IFN-alpha) plus ribavirin is used to treat hepatitis C virus (HCV) infection and is associated with a high rate of depression. Newer, pegylated preparations of IFN-alpha have a longer half-life, require once-per-week dosing, and may be associated with reduced neuropsychiatric burden. Limited data exist on depression during pegylated IFN-alpha therapy. Depressive symptoms were assessed using the Zung Self-Rating Depression Scale (SDS) in 162 HCV-infected patients at baseline and after 4, 8, 12, and 24 weeks of treatment with pegylated IFN alpha-2b (PEG IFN) plus weight-based (N = 86) versus standard dose (N = 76) ribavirin. Data were collected from March 2001 to April 2003. Compared with baseline, mean SDS index scores were significantly increased by week 4 and remained elevated throughout the study. Thirty-nine percent of the sample experienced moderate to severe depressive symptoms (SDS index score > or = 60) at some point during PEG IFN/ribavirin therapy. Baseline depression scores significantly predicted severity of depressive symptoms during PEG IFN/ribavirin treatment (simple regression analysis: Y = 0.55X + 32.7, p < .0001). In addition, assignment to weight-based ribavirin treatment and history of depression were associated with increased likelihood of developing moderate to severe depressive symptoms (odds ratio [OR] = 2.7, 95% CI = 1.3 to 5.6, p < .01, and OR = 3.3, 95% CI = 1.3 to 8.1, p < .01, respectively). Development of moderate to severe depressive symptoms occurred frequently during PEG IFN/ribavirin treatment and was predicted by baseline depression scores and higher doses of ribavirin. History of major depressive disorder was also a significant predictive factor, but only through association with elevated baseline depression status. All of these factors can be evaluated and addressed to limit neuropsychiatric morbidity during HCV treatment.


Synthesis of Nitrogen-Doped Graphene Quantum Dots at Low Temperature for Electrochemical Sensing Trinitrotoluene.
Nitrogen-doped graphene quantum dots (N-GQDs) are synthesized at low temperature as a new catalyst allowing electrochemical detection of 2,4,6-trinitrotoluene (TNT). N-GQDs are made by an oxidative ultrasonication of graphene oxide (GO) forming nanometer-sized species, which are then chemically reduced and nitrogen doped by reacting with hydrazine. The as-synthesized N-GQDs have an average diameter of ∼2.5 nm with an N/C atomic ratio of up to ∼6.4%. To detect TNT, TNT is first accumulated on N-GQDs modified glassy carbon (N-GQDs/GC) electrode by holding the electrode at a 0 V versus Ag/AgCl for 150 s in an aqueous TNT solution. Next, the N-GQDs/GC electrode with accumulated TNT is transferred to a fresh PBS solution (0.1 M, pH 7.0, without TNT), where the TNT reduction current at -0.36 V versus Ag/AgCl in a linear scan voltammogram (LSV) shows a linear response to TNT concentration in the aqueous solution from 1 to 400 ppb, with a correlation coefficient of 0.999, a detection limit of 0.2 ppb at a signal/noise (S/N) of 3, and a detection sensitivity of 363 ± 7 mA mM(-1) cm(-2). The detection limit of 0.2 ppb of TNT for this new method is much lower than 2 ppb set by the U.S. Environmental Protection Agency for drinking water. Therefore, N-GQDs allow an electrochemical method for assaying TNT in drinking water to determine if levels of TNT are safe or not.


Clinical Presentation and Features of Juvenile-Onset Huntington's Disease: A Systematic Review.
Juvenile-onset Huntington's disease (JHD) is defined by onset at the age of 20 or younger and represents approximately 5% of all HD cases. Patients with JHD present with a broad range of symptoms and signs that only overlap partially with adult-onset HD. A greater awareness and understanding of the presentation of JHD would improve the diagnosis and treatment of this condition. To undertake a systematic review of the literature relating to the clinical features at first presentation of JHD. We searched MEDLINE and EMBASE for all studies describing presenting features of JHD patients, performed quality control, and collated and analysed the data. We screened 2917 records for eligibility, and included 79 studies (n = 285 individuals) in the analysis. All were case reports and case series, synthesising data from 25 different countries. Thirty-four different clinical features at presentation were identified. Four groups of symptoms or signs were present in more than 15% of cases: behavioural disturbance, falls/gait disturbance, cognitive impairment and parkinsonian features. Where data were available, the median age of onset was 9 years, 52% were female, the mutant HTT allele was transmitted paternally in 80% of cases, and the median CAG repeat length was 64. JHD can present with a wide variety of symptoms and signs, with non-motor characteristics being observed most frequently. Greater recognition of these presentations will facilitate early diagnosis and management. Tailored rating scales to score motor, non-motor, and functional impairments specifically in JHD are required to standardise research studies, and are under development.


The impact of hypertension and nicotine on the size of ruptured intracranial aneurysms.
The goal of the study was to analyse retrospectively the impact of risk factors for subarachnoid haemorrhage (SAH) on the size of ruptured intracranial aneurysms in order to identify variables that might influence the discrepancy between average sizes of ruptured and unruptured aneurysms. The records and angiographies of 373 patients treated at our centre due to aneurysmal SAH between 2004 and December 2008 were retrospectively analysed. Modifiable and non-modifiable risk factors for SAH were correlated with exact measurement of aneurysm size using three-dimensional rotational digital subtraction angiographies (3D-DSA). Average maximum aneurysm diameter in patients with combined history of hypertension and cigarette smoking was 5.47±3.22 mm (95% CI 4.71 to 6.24); thus, significantly smaller (p<0.001) than in patients with hypertension only (6.27±3.28 mm, 95% CI 5.75 to 6.78), with cigarette smoking only (7.61±4.29 mm, 95% CI 6.43 to 8.79) and patients with no history for risk factors (8.08±4.73 mm, 95% CI 6.96 to 9.21). Odds ratio (OR) for aneurysm size less than 7 mm in patients with combined hypertension and cigarette smoking was 3.63 (95% CI 1.78 to 7.42), 3.09 (95% CI 1.95 to 4.92) in patients with hypertension only and 1.02 (95% CI 0.64 to 1.62) in patients with cigarette smoking only. The present analysis demonstrates a distinct correlation between hypertension, smoking and the size of ruptured aneurysms in SAH patients. Arterial hypertension and cigarette smoking appear to destabilise cerebral aneurysms' growth. Our data strongly suggest that these factors should also be considered when treatment indications for small unruptured aneurysms are discussed.


The Drosophila shark tyrosine kinase is required for embryonic dorsal closure.
Dorsal closure (DC) in the Drosophila embryo requires the coordinated interaction of two different functional domains of the epidermal cell layer-the leading edge (LE) and the lateral epidermis. In response to activation of a conserved c-Jun amino-terminal kinase (JNK) signaling module, the dorsal-most layer of cells, which constitute the LE of the stretching epithelial sheet, secrete Dpp, a member of the TGFbeta superfamily. Dpp and other LE cell-derived signaling molecules stimulate the bilateral dorsal elongation of cells of the dorsolateral epidermis over the underlaying amnioserosa and the eventual fusion of their LEs along the dorsal midline. We have found that flies bearing a Shark tyrosine kinase gene mutation, shark(1), exhibit a DC-defective phenotype. Dpp fails to be expressed in shark(1) mutant LE cells. Consistent with these observations, epidermal-specific reconstitution of shark function or overexpression of an activated form of c-Jun in the shark(1) mutant background, rescues the DC defect. Thus, Shark regulates the JNK signaling pathway leading to Dpp expression in LE cells. Furthermore, constitutive activation of the Dpp pathway throughout the epidermis fails to rescue the shark(1) DC defect, suggesting that Shark may function in additional pathways in the LE and/or lateral epithelium.


Simultaneous enantiospecific recognition of several β-blocker enantiomers using molecularly imprinted polymer-based electrochemical sensor.
The development of a chiral electrochemical sensor using an electrogenerated molecularly imprinted polymer (MIP)-based ultrathin film using R(+)-atenolol (ATNL) as a template was reported. The proposed sensor exhibited distinctive enantiospecific oxidation peaks toward the R-antipodes of four β-blocker representatives and additional oxidation peaks common to both enantiomers of each studied β-blocker, allowing thus the simultaneous analysis of all of their enantiomers in a single analysis. The specific preconditioning of the polymer by alternative exposure to aqueous and nonaqueous medium was proven to be essential for the chiral recognition ability of the obtained sensor. The rebinding property of the MIP film was studied by using a well-known redox probe, a change in the morphology and diffusive permeability of the thin polymeric layer in the presence of its template being observed. The applicability of the optimized analytical procedure was demonstrated by the analysis of ATNL's enantiomers in the range of 1.88 × 10(-7)-1.88 × 10(-5) mol/L. The developed polymeric interface is the first reported transductor of a chiral electrochemical sensor able to exhibit simultaneous enantiospecificity toward several β-blocker representatives extensively used in the pharmaceutical and biomedical fields, offering good prospects in the simple, cost-effective, and fast assessment of their enantiomeric ratio and total concentration.


Relevance of internal time and circadian robustness for cancer patients.
Adequate circadian timing of cancer treatment schedules (chronotherapy) can enhance tolerance and efficacy several-fold in experimental and clinical situations. However, the optimal timing varies according to sex, genetic background and lifestyle. Here, we compute the individual phase of the Circadian Timing System to decipher the internal timing of each patient and find the optimal treatment timing. Twenty-four patients (11 male; 13 female), aged 36 to 77 years, with advanced or metastatic gastro-intestinal cancer were recruited. Inner wrist surface Temperature, arm Activity and Position (TAP) were recorded every 10 min for 12 days, divided into three 4-day spans before, during and after a course of a set chronotherapy schedule. Pertinent indexes, I < O and a new biomarker, DI (degree of temporal internal order maintenance), were computed for each patient and period. Three circadian rhythms and the TAP rhythm grew less stable and more fragmented in response to treatment. Furthermore, large inter- and intra-individual changes were found for T, A, P and TAP patterns, with phase differences of up to 12 hours among patients. A moderate perturbation of temporal internal order was observed, but the administration of fixed chronomodulated chemotherapy partially resynchronized temperature and activity rhythms by the end of the study. The integrated variable TAP, together with the asynchrony among rhythms revealed by the new biomarker DI, would help in the personalization of cancer chronotherapy, taking into account individual circadian phase markers.


Effects of ERβ and ERα on OVX-induced changes in adiposity and insulin resistance.
Loss of ovarian hormones leads to increased adiposity and insulin resistance (IR), increasing the risk for cardiovascular and metabolic diseases. The purpose of this study was to investigate whether the molecular mechanism behind the adverse systemic and adipose tissue-specific metabolic effects of ovariectomy requires loss of signaling through estrogen receptor alpha (ERα) or estrogen receptor β (ERβ). We examined ovariectomized (OVX) and ovary-intactwild-type (WT), ERα-null (αKO), and ERβ-null (βKO) female mice (age ~49 weeks; n = 7-12/group). All mice were fed a phytoestrogen-free diet (<15 mg/kg) and either remained ovary-intact (INT) or were OVX and followed for 12 weeks. Body composition, energy expenditure, glucose tolerance, and adipose tissue gene and protein expression were analyzed. INT αKO were ~25% fatter with reduced energy expenditure compared to age-matched INT WT controls and βKO mice (all P < 0.001). Following OVX, αKO mice did not increase adiposity or experience a further increase in IR, unlike WT and βKO, suggesting that loss of signaling through ERα mediates OVX-induced metabolic dysfunction. In fact, OVX in αKO mice (i.e., signaling through ERβ in the absence of ERα) resulted in reduced adiposity, adipocyte size, and IR (P < 0.05 for all). βKO mice responded adversely to OVX in terms of increased adiposity and development of IR. Together, these findings challenge the paradigm that ERα mediates metabolic protection over ERβ in all settings. These findings lead us to suggest that, following ovarian hormone loss, ERβ may mediate protective metabolic benefits.


The genome of swinepox virus.
Swinepox virus (SWPV), the sole member of the Suipoxvirus genus of the Poxviridae, is the etiologic agent of a worldwide disease specific for swine. Here we report the genomic sequence of SWPV. The 146-kbp SWPV genome consists of a central coding region bounded by identical 3.7-kbp inverted terminal repeats and contains 150 putative genes. Comparison of SWPV with chordopoxviruses reveals 146 conserved genes encoding proteins involved in basic replicative functions, viral virulence, host range, and immune evasion. Notably, these include genes with similarity to genes for gamma interferon (IFN-gamma) receptor, IFN resistance protein, interleukin-18 binding protein, IFN-alpha/beta binding protein, extracellular enveloped virus host range protein, dUTPase, hydroxysteroid dehydrogenase, superoxide dismutase, serpin, herpesvirus major histocompatibility complex inhibitor, ectromelia virus macrophage host range protein, myxoma virus M011L, variola virus B22R, four ankyrin repeat proteins, three kelch-like proteins, five vaccinia virus (VV) A52R-like family proteins, and two G protein-coupled receptors. The most conserved genomic region is centrally located and corresponds to the VV region located between genes F9L and A38L. Within the terminal 13 kbp, colinearity is disrupted and multiple poxvirus gene homologues are absent or share a lower percentage of amino acid identity. Most of these differences involve genes and gene families with likely functions involving viral virulence and host range. Three open reading frames (SPV018, SPV019. and SPV020) are unique for SWPV. Phylogenetic analysis, genome organization, and amino acid identity indicate that SWPV is most closely related to the capripoxvirus lumpy skin disease virus, followed by the yatapoxvirus yaba-like disease virus and the leporipoxviruses. The gene complement of SWPV better defines Suipoxvirus within the Chordopoxvirinae subfamily and provides a basis for future genetic comparisons.


Fasting plasma glucose levels predict steroid-induced abnormal glucose metabolism in patients with non-diabetic chronic kidney disease: a prospective cohort study.
Glucocorticoids-induced abnormal glucose metabolism (AGM) is a common medical problem in patients with non-diabetic chronic kidney disease (CKD). However, little information is available regarding the prediction of steroid-induced AGM in this patient population. In this prospective cohort study, we consecutively enrolled 303 non-diabetic CKD patients with fasting plasma glucose (FPG) levels <5.6 mmol/l and normal oral glucose tolerance test (OGTT). OGTT was performed every 3 months during glucocorticoid treatment to identify new-onset AGM, and patients were followed for 12 months post steroid withdrawal. During 593 person-years, there were 107 incident cases of steroid-induced AGM (18/100 person-year), including 55 (51.4%) diabetes and 52 (48.6%) pre-diabetes. In a multivariate model, each millimole increase per liter in FPG enhanced the risk of AGM by 4.6-fold (hazard ratio 4.58, 95% confidence interval, 2.67-7.83). After adjusting other risk factors, a progressively increased risk of AGM or DM was observed in patients with FPG levels ≥4.8 mmol/l, as compared with those whose levels were ≤4.3 mmol/l (p for trend <0.001). Furthermore, a greater increase in FPG level (≥0.3 mmol/l) during the first 3 months of glucocorticoid treatment was associated with an increased risk for future diabetes. For predicting steroid-induced diabetes, the area under the receiver-operating characteristic curve was 0.90 for the combination of FPG and changes in FPG levels at month 3. Higher-normal FPG and a greater increase in FPG levels during glucocorticoid treatment may help to identify non-diabetic CKD patients at increased risk of steroid-induced AGM or diabetes.


The hepatic circadian clock modulates xenobiotic metabolism in mice.
The circadian clock generates daily cycles of gene expression that regulate physiological processes. The liver plays an important role in xenobiotic metabolism and also has been shown to possess its own cell-based clock. The liver clock is synchronized by the master clock in the brain, and a portion of rhythmic gene expression can be driven by behavior of the organism as a whole even when the hepatic clock is suppressed. So far, however, there is relatively little evidence indicating whether the liver clock is functionally important in modulating xenobiotic metabolism. Thus, mice lacking circadian clock function in the whole body or specifically in liver were challenged with pentobarbital and acetaminophen, and pentobarbital sleep time (PBST) and acetaminophen toxicity, respectively, was assessed at different times of day in mutant and control mice. The results suggest that the liver clock is essential for rhythmic changes in xenobiotic detoxification. Surprisingly, it seems that the way in which the clock is disrupted determines the rate of xenobiotic metabolism in the liver. CLOCK-deficient mice are remarkably resistant to acetaminophen and exhibit a longer PBST, while PERIOD-deficient mice have a short PBST. These results indicate an essential role of the tissue-intrinsic peripheral circadian oscillator in the liver in regulating xenobiotic metabolism.


Polarization-multiplexed encoding at nanometer scales.
Optical data storage was developed using binary encoding primarily due to signal to noise ratio considerations. We report on a multiplexing method that allows a seven fold storage increase, per storage layer, per side, and propose one that can yield theoretically a 20+ fold increase. Multiplexing is achieved by encoding information in polarization via appropriately oriented nanostructures that emit strongly polarized light when excited by unpolarized light. The storage increase is possible due to the significantly reduced crosstalk that results form using unpolarized light.


EGFR-dependent network interactions that pattern Drosophila eggshell appendages.
Similar to other organisms, Drosophila uses its Epidermal Growth Factor Receptor (EGFR) multiple times throughout development. One crucial EGFR-dependent event is patterning of the follicular epithelium during oogenesis. In addition to providing inductive cues necessary for body axes specification, patterning of the follicle cells initiates the formation of two respiratory eggshell appendages. Each appendage is derived from a primordium comprising a patch of cells expressing broad (br) and an adjacent stripe of cells expressing rhomboid (rho). Several mechanisms of eggshell patterning have been proposed in the past, but none of them can explain the highly coordinated expression of br and rho. To address some of the outstanding issues in this system, we synthesized the existing information into a revised mathematical model of follicle cell patterning. Based on the computational model analysis, we propose that dorsal appendage primordia are established by sequential action of feed-forward loops and juxtacrine signals activated by the gradient of EGFR signaling. The model describes pattern formation in a large number of mutants and points to several unanswered questions related to the dynamic interaction of the EGFR and Notch pathways.


Stroma: fertile soil for inflammation.
Biological therapies for the management of immune mediated inflammatory diseases such as rheumatoid arthritis have proven to be extremely successful in recent years. Despite these successes, even the most effective of therapies do not lead to cure. Why chronic inflammation persists indefinitely within the rheumatoid synovium despite an absence of continuous stimulation, and why some patients with early synovitis progress to persistent disease whilst others do not, has remained unexplained. In contrast to the paradigm that stromal cells are biochemically active but immunologically passive, there is now growing evidence that stromal components from the rheumatoid synovium play a crucial part in the immunopathology of rheumatoid arthritis. Stromal cells play a central role in the transformation of an acute, resolving to a chronic inflammatory process, and to the persistence of synovial inflammation and joint destruction through a variety of immune mechanisms. Therapeutic manipulation of the stroma is a largely unexplored, yet potentially vital area of research. Targeting pathogenic stromal cells has the potential to provide a cure for chronic inflammatory disorders such as rheumatoid arthritis.


Facial skin tightening with an 1100-1800 nm infrared device.
To determine the efficacy of the 1100-1800 nm infrared device for facial and cervical skin tightening. Ten female patients, with a mean age of 56.5 years, received two treatments 1 month apart with a chilled tip infrared device (Titan; Cutera, Brisbane, CA, USA). Individuals were examined and photographed prior to treatment and at 1 and 3 months post-treatment. Three treatment-independent evaluators compared the photographs and graded them on a standardized scale applied to seven regions subdividing the face and neck. After evaluating the photographs, the difference in pretreatment and post-treatment scores was expressed as a percentage. The patients also rated their results. The greatest tightening was achieved over the malar region, the upper neck and the body of the mandible. In these areas the average tightening was 10%, 10%, and 12% respectively. The patients reported a 32% improvement in the appearance of their cheeks and a 20% visible improvement in their necks. Overall, they were pleased with the result of this non-surgical skin tightening.


Lipid management in chronic kidney disease: synopsis of the Kidney Disease: Improving Global Outcomes 2013 clinical practice guideline.
The Kidney Disease: Improving Global Outcomes (KDIGO) organization developed a clinical practice guideline in 2013 on lipid management and treatment of all adults and children with chronic kidney disease (CKD). All forms of CKD are included (non-dialysis-dependent, dialysis-dependent, and kidney transplant recipients). The KDIGO Lipid Guideline Development Work Group defined the scope of the guideline, gathered evidence, determined topics for systematic review, and graded the quality of evidence that had been summarized by an evidence review team. Searches of the English-language literature were conducted through August 2011 and supplemented by targeted searches through June 2013. Final modification of the guidelines was informed by the KDIGO Board of Directors and a public review process involving registered stakeholders. The full guideline includes 13 recommendations; a key element was the recommendation for statin or statin with ezetimibe treatment of adults aged 50 years or older with estimated glomerular filtration rates less than 60 mL/min/1.73 m2 but not treated with long-term dialysis or kidney transplantation. This synopsis focuses on 8 recommendations pertinent to assessment of lipid status and treatment with a statin-based regimen in adults.


Comminuted radial head fractures treated with a modular metallic radial head arthroplasty. Study of outcomes.
Comminuted fractures of the radial head are challenging to treat with open reduction and internal fixation. Radial head arthroplasty is an alternative treatment with results that compare favorably with those reported after open reduction and internal fixation of similar fractures. The purpose of this study was to evaluate the two-year outcomes and the rate of recovery of a closely followed cohort of patients in whom an unreconstructible radial head fracture had been treated with a modular metallic prosthesis. Twenty-six patients (seventeen female and nine male; mean age, fifty-four years) with an unreconstructible comminuted radial head fracture and associated elbow injuries were treated with a modular metallic radial head arthroplasty. Patients who had presented more than four weeks following the injury or had had the radial head arthroplasty as a second-stage or salvage procedure were excluded. Of the twenty-six patients, twenty-two had an associated elbow dislocation, and thirteen of them also had an associated fracture of the coronoid process. Patients were prospectively followed at three, six, twelve, and twenty-four months. Self-reported limb function, general health, range of motion, and isometric strength were assessed by an independent observer. Following treatment of the injury, significant decreases in self-reported and measured impairments were noted over time, with the majority of the recovery occurring by six months and little further recovery noted between six and twenty-four months. There were slight-to-moderate deficits in the range of motion and strength compared with the values on the contralateral, unaffected side. Patient satisfaction was high at three months and remained high at two years. All elbow joints remained stable, no implant required revision, and there was no evidence of overstuffing of the joint. Mild osteoarthritis was seen in five (19%) of the twenty-six patients. An arthroplasty with a modular metallic radial head is a safe and effective option for the treatment of unreconstructible radial head fractures associated with other elbow injuries. Recovery primarily occurs by six months, with minimal additional improvements over the next eighteen months.


Muscle progenitor cells failing to respond to positional cues adopt non-myogenic fates in myf-5 null mice.
Mice that have mutations in both myogenic transcription factors Myf-5 and MyoD totally lack skeletal muscle fibres and their precursor myoblasts, whereas with either mutation alone, muscle is present. Skeletal muscle in the vertebrate body is derived from epithelial somites that respond to environmental signals to form the dorsal epithelial dermomyotome (dermis, muscle) and ventral mesenchymal sclerotome (axial skeleton, ribs). The first muscle, the myotome, forms centrally in the somite, when only myf-5 is programming myogenesis. By targeting the nlacZ reporter gene into the myf-5 locus, we demonstrate that beta-galactosidase+ muscle progenitor cells are present in the dermomyotome of myf-5 null embryos, and that they undergo a normal epithelial-mesenchymal transition; however, they migrate aberrantly. Dorsally, they accumulate under the ectoderm and express a non-muscle dermal marker, Dermo-1. Ventrally, beta-galactosidase+ cells also fail to localize correctly, express a cartilage marker scleraxis, and are subsequently found in ribs. Therefore Myf-5 protein is necessary for cells to respond correctly to positional cues in the embryo and to adopt their myogenic fate. In its absence, muscle progenitors, having activated myf-5, remain multipotent and differentiate into other somitic derivatives according to their local environment.


Neurophysiologic intraoperative monitoring (NIOM) in pediatric patients with polyneuropathy.
Neurophysiologic intraoperative monitoring (NIOM) abnormalities during scoliosis surgery led to a diagnosis of Friedreich's ataxia in this illustrative case. This prompted the retrospective examination of NIOM for pediatric scoliosis surgery in polyneuropathy patients. Among patients who underwent scoliosis surgery in 2010-2017, there were six polyneuropathy patients identified. Their clinical history and baseline NIOM data were reviewed. Scoliosis accompanied Charcot-Marie-Tooth disease, Friedreich's ataxia, and ataxia telangiectasia. Some patients with no recorded somatosensory evoked potentials (SEPs) exhibited motor evoked potentials (MEPs); no patients with absent MEPs had SEPs present. NIOM modifications included SEP stimulation rate; type of SEP electrodes used; train parameters for MEP acquisition; and sweep speed. This sample of NIOM data for previously monitored scoliosis cases in children with polyneuropathy allowed investigation of patterns of findings and troubleshooting attempts to optimize monitoring. Attentiveness to pertinent medical history prepared the NIOM team to change typical recording parameters based on underlying polyneuropathy. A multimodality approach provided useful information as several of these cases would have been unmonitorable with use of SEPs alone. As for the case described, the awareness of NIOM patterns in polyneuropathy may guide evaluations of patients with presumed idiopathic scoliosis who have unrecognized polyneuropathy.


In vitro and in vivo antimicrobial activity of combined therapy of silver nanoparticles and visible blue light against Pseudomonas aeruginosa.
Silver nanoparticles (AgNPs) have been used as potential antimicrobial agents against resistant pathogens. We investigated the possible therapeutic use of AgNPs in combination with visible blue light against a multidrug resistant clinical isolate of Pseudomonas aeruginosa in vitro and in vivo. The antibacterial activity of AgNPs against P. aeruginosa (1×10(5) colony forming unit/mL) was investigated at its minimal inhibitory concentration (MIC) and sub-MIC, alone and in combination with blue light at 460 nm and 250 mW for 2 hours. The effect of this combined therapy on the treated bacteria was then visualized using transmission electron microscope. The therapy was also assessed in the prevention of biofilm formation by P. aeruginosa on AgNP-impregnated gelatin biopolymer discs. Further, in vivo investigations were performed to evaluate the efficacy of the combined therapy to prevent burn-wound colonization and sepsis in mice and, finally, to treat a real infected horse with antibiotic-unresponsive chronic wound. The antimicrobial activity of AgNPs and visible blue light was significantly enhanced (P<0.001) when both agents were combined compared to each agent alone when AgNPs were tested at MIC, 1/2, or 1/4 MIC. Transmission electron microscope showed significant damage to the cells that were treated with the combined therapy compared to other cells that received either the AgNPs or blue light. In addition, the combined treatment significantly (P<0.001) inhibited biofilm formation by P. aeruginosa on gelatin discs compared to each agent individually. Finally, the combined therapy effectively treated a horse suffering from a chronic wound caused by mixed infection, where signs of improvement were observed after 1 week, and the wound completely healed after 4 weeks. To our knowledge, this combinatorial therapy has not been investigated before. It was proved efficient and promising in managing infections caused by multidrug resistant bacteria and could be used as an alternative to conventional antibiotic therapy.


Life-threatening flecainide toxicity. A pharmacodynamic approach.
Ventricular tachyarrhythmias and severe bradycardia, flecainide acetate's most serious side effects, occur in patients with chronic heart disease or in healthy persons after frank overdose. Toxic effects appear to correlate closely with drug levels in plasma. The surface electrocardiogram can indicate toxicity by demonstrating this drug's electrophysiologic potency to depress all major cardiac conduction pathways with marked prolongation of the PR and QRS intervals. We report the clinical course of a young, healthy person who developed polymorphous ventricular tachycardia after taking a 3800-mg dose of flecainide acetate. Sinus rhythm was restored without pacing or cardioversion after infusion of a beta-sympathomimetic agent, physostigmine, and a sodium load. The initial flecainide serum levels were five times greater than the usual upper therapeutic level. Electrocardiography showed that the lengthened electrocardiographic time intervals decreased in correlation with falling drug levels. Flecainide-associated life-threatening arrhythmias and available therapeutic interventions are discussed.


Neuromuscular and circulatory adaptation during combined arm and leg exercise with different maximal work loads.
Cardiopulmonary kinetics and electromyographic activity (EMG) during exhausting exercise were measured in 8 males performing three maximal combined arm+leg exercises (cA+L). These exercises were performed at different rates of work (mean+/-SD; 373+/-48, 429+/-55 and 521+/-102 W) leading to different average exercise work times in all tests and subjects. VO2 reached a plateau versus work rate in every maximal cA+L exercise (range 6 min 33 s to 3 min 13 s). The three different exercise protocols gave a maximal oxygen consumption (VO2MAX) of 4.67+/-0.57, 4.58+/-0.52 and 4.66+/-0.53 l min(-1) (P=0.081), and a maximal heart rate (HRmax) of 190+/-6, 189+/-4 and 189+/-6 beats min(-1) (P=0.673), respectively. Root mean square EMG (EMGRMS) of the vastus lateralis and the triceps brachii muscles increased with increasing rate of work and time in all three cA+L protocols. The study demonstrates that despite different maximal rates of work, leading to different times to exhaustion, the circulatory adaptation to maximal exercise was almost identical in all three protocols that led to a VO2 plateau. The EMG(RMS) data showed increased muscle recruitment with increasing work rate, even though the HRmax and VO2MAX was the same in all three cA+L protocols. In conclusion, these findings do not support the theory of the existence of a central governor (CG) that regulates circulation and neuronal output of skeletal muscles during maximal exercise.


Bronchial epithelial innate and adaptive immunity signals are induced by polycyclic aromatic hydrocarbons.
Polycyclic aromatic hydrocarbons including Benzo[a]pyrene have been recognised as important pollutant chemicals with the potential to influence the respiratory system in disease. Airway epithelial cells are an integral component of how immune responses are directed as a consequence of exposure to inhaled material. It was aim of this study to examine how such cells respond to PAH exposure and to characterise the immune response. Human primary bronchial epithelial cells (HPBECs) were exposed to Benzo[a]pyrene, Benzo[e]pyrene, Fluoranthene and Benzo[b]fluoranthene for 24 h and a repeat exposure up to 7 days, and examined for global gene expression using RNA-Seq. In addition to increased expression of CYP1A1 and other AHR dependent changes, we identified significant increases in innate and adaptive immune signals including, IL-1A, IL-19, SERPINB2, STAT6, HLA-DMB and HLA-DRA. We also observed increased expression of HMOX1 and NQO1, genes involved in the response to oxidative stress. Immune system related gene expression was differentially induced by each compound with Benzo[a]pyrene and Benzo[b]fluoranthene demonstrating the most potent responses. Differential induction paralleled the level to which AHR dependent gene expression and oxidative stress markers were induced. We also observed similar levels of gene expression when cells were exposed to organic extracts from diesel exhaust particles. In conclusion, hazard characterisation of responses to PAH exposure in HPBECs highlights specific responses of both innate and adaptive immunity.


Diurnal variation in suicide timing by age and gender: Evidence from Japan across 41 years.
Previous research on hourly diurnal variation in suicide frequency has often suffered from geographical and time-span limitations in the data. We studied diurnal and daily variations of suicide by analyzing a large dataset based on the national death registry in Japan between 1974 and 2014. The diurnal and daily patterns of 873,268 suicide deaths over 41 years were examined by sex and age group through Poisson regression and visual inspection. We also investigated whether these patterns are related to Japan's economic conditions. Suicide by middle-aged males was most frequent in the early morning especially on Mondays after the end of Japan's high growth period. We also observed large midnight peaks in suicides among young and middle-aged males. The proportion of early morning suicide deaths by young and middle-aged males increased as the country's unemployment rose. Females and elderly males were more likely to die by suicide during the day than at night. Our study examined time of death, not time of suicide attempt. It is possible that there is a discrepancy between the two. Different subpopulations die by suicide at different times of the day and days of the week. Time patterns of suicide varied considerably over time, suggesting that they cannot be explained by biological circadian rhythm alone. Our findings suggest that the patterns are partly explained by economic conditions. Future suicide prevention efforts should consider the time patterns of suicide unique to each subpopulation, especially when economic growth is depressed.


Molecular epidemiology and host genetics of norovirus and rotavirus infections in Portuguese elderly living in aged care homes.
Norovirus (NoV) and rotavirus group A (RVA) are major agents of acute gastroenteritis worldwide. This study aimed to investigate their epidemiological profile in Portuguese elderly living in long-term care facilities and to assess the host genetic factors mediating infection susceptibility. From November 2013 to June 2015, 636 faecal specimens from 169 elderly, mainly asymptomatic, living in nursing homes in Greater Lisbon and Faro district, Portugal, were collected. NoV and RVA were detected by real-time polymerase chain reaction and NoV genotyped by phylogenetic analysis. NoV detection rate was 7.1% (12 of 169). Three GI.3 and one GII.6 strains were genotyped. RVA detection rate was 3.6% (6 of 169), exclusively in asymptomatic individuals. Host genetic factors associated with infection susceptibility were described on 250 samples by saliva-based enzyme-linked immunosorbent assays. The Lewis-negative phenotype was 8.8% (22 of 250) and the rate of nonsecretors was 16.8% (42 of 250). Association to NoV and RVA infection was performed in the subgroup of individuals (n = 147) who delivered both faecal and saliva samples. The majority of NoV- and RVA-positive individuals (90.9% and 83.3%, respectively) were secretor-positive, with Lewis B phenotype. In a subset of individuals, FUT2 and FUT3 genes were genotyped to assess mutations and validate the secretor and Lewis phenotypes. All sequenced nonsecretors were homozygous for FUT2 nonsense mutation G428A. In this study, low detection rates of NoV and RVA infections were found during two winter seasons. However, even in the absence of any outbreak, the importance of finding these infections in a nonepidemic situation in long-term care facilities may have important implications for infection control.


Herbivorous protist growth and grazing rates at in situ and artificially elevated temperatures during an Arctic phytoplankton spring bloom.
To assess protistan grazing impact and temperature sensitivity on plankton population dynamics, we measured bulk and species-specific phytoplankton growth and herbivorous protist grazing rates in Disko Bay, West Greenland in April-May 2011. Rate estimates were made at three different temperatures in situ (0 °C), +3 °C and +6 °C over ambient. In situ Chlorophyll a (Chl a) doubled during the observation period to ∼12 µg Chl a L-1, with 60-97% of Chl a in the >20 µm size-fraction dominated by the diatom genus Chaetoceros. Herbivorous dinoflagellates comprised 60-80% of microplankton grazer biomass. At in situ temperatures, phytoplankton growth or grazing by herbivorous predators <200 µm was not measurable until 11 days after observations commenced. Thereafter, phytoplankton growth was on average 0.25 d-1. Phytoplankton mortality due to herbivorous grazing was only measured on three occasions but the magnitude was substantial, up to 0.58 d-1. Grazing of this magnitude removed ∼100% of primary production. In short-term temperature-shift incubation experiments, phytoplankton growth rate increased significantly (20%) at elevated temperatures. In contrast, herbivorous protist grazing and species-specific growth rates decreased significantly (50%) at +6 °C. This differential response in phytoplankton and herbivores to temperature increases resulted in a decrease of primary production removed with increasing temperature. Phaeocystis spp. abundance was negatively correlated with bulk grazing rate. Growth and grazing rates were variable but showed no evidence of an inherent, low temperature limitation. Herbivorous protist growth rates in this study and in a literature review were comparable to rates from temperate waters. Thus, an inherent physiological inhibition of protistan growth or grazing rates in polar waters is not supported by the data. The large variability between lack of grazing and high rates of primary production removal observed here and confirmed in the literature for polar waters implies larger amplitude fluctuations in phytoplankton biomass than slower, steady grazing losses of primary production.


Stimulant reduction intervention using dosed exercise (STRIDE) - CTN 0037: study protocol for a randomized controlled trial.
There is a need for novel approaches to the treatment of stimulant abuse and dependence. Clinical data examining the use of exercise as a treatment for the abuse of nicotine, alcohol, and other substances suggest that exercise may be a beneficial treatment for stimulant abuse, with direct effects on decreased use and craving. In addition, exercise has the potential to improve other health domains that may be adversely affected by stimulant use or its treatment, such as sleep disturbance, cognitive function, mood, weight gain, quality of life, and anhedonia, since it has been shown to improve many of these domains in a number of other clinical disorders. Furthermore, neurobiological evidence provides plausible mechanisms by which exercise could positively affect treatment outcomes. The current manuscript presents the rationale, design considerations, and study design of the National Institute on Drug Abuse (NIDA) Clinical Trials Network (CTN) CTN-0037 Stimulant Reduction Intervention using Dosed Exercise (STRIDE) study. STRIDE is a multisite randomized clinical trial that compares exercise to health education as potential treatments for stimulant abuse or dependence. This study will evaluate individuals diagnosed with stimulant abuse or dependence who are receiving treatment in a residential setting. Three hundred and thirty eligible and interested participants who provide informed consent will be randomized to one of two treatment arms: Vigorous Intensity High Dose Exercise Augmentation (DEI) or Health Education Intervention Augmentation (HEI). Both groups will receive TAU (i.e., usual care). The treatment arms are structured such that the quantity of visits is similar to allow for equivalent contact between groups. In both arms, participants will begin with supervised sessions 3 times per week during the 12-week acute phase of the study. Supervised sessions will be conducted as one-on-one (i.e., individual) sessions, although other participants may be exercising at the same time. Following the 12-week acute phase, participants will begin a 6-month continuation phase during which time they will attend one weekly supervised DEI or HEI session. ClinicalTrials.gov, NCT01141608 http://clinicaltrials.gov/ct2/show/NCT01141608?term=Stimulant+Reduction+Intervention+using+Dosed+Exercise&rank=1.


Adjusting for bias introduced by instrumental variable estimation in the Cox proportional hazards model.
Instrumental variable (IV) methods are widely used for estimating average treatment effects in the presence of unmeasured confounders. However, the capability of existing IV procedures, and most notably the two-stage residual inclusion (2SRI) algorithm recommended for use in non-linear contexts, to account for unmeasured confounders in the Cox proportional hazard model is unclear. We show that instrumenting an endogenous treatment induces an unmeasured covariate, referred to as an individual frailty in survival analysis parlance, which if not accounted for leads to bias. We propose a new procedure that augments 2SRI with an individual frailty and prove that it is consistent under certain conditions. The finite sample-size behavior is studied across a broad set of conditions via Monte Carlo simulations. Finally, the proposed methodology is used to estimate the average effect of carotid endarterectomy versus carotid stenting on the mortality of patients suffering from carotid artery disease. Results suggest that the 2SRI-frailty estimator generally reduces the bias of both point and interval estimators compared to traditional 2SRI.


An Indoor Therapeutic Garden for Behavioral Symptoms in Alzheimer's Disease: A Randomized Controlled Trial.
Behavioral and psychological symptoms of dementia (BPSD) affect 60-90% of patients with Alzheimer's disease (AD). To determine if environmental therapy is an effective strategy to reduce BPSD, we tested 163 patients with AD with Neuropsychiatric Inventory (NPI) before and after 6 months of an indoor therapeutic garden (TG) or standard environment. A single-blind randomized controlled trial on AD patients with BPSD. Participants were randomized to an indoor TG (N = 82), or standard environment (control, N = 81) for 6 months. change in the NPI score from baseline (T0) to end of treatment (T1). change in use of quetiapine, cognition, activities of daily living, salivary cortisol, blood pressure from T0 to T1. NPI score significantly ameliorated (TG versus control: -31.8 points), quetiapine dosage (-150 mg), blood pressure (-2.6 mm Hg), and salivary cortisol (-6.4 to -2.1 Nmol/l) were significantly reduced, the Mini-Mental State Examination significantly improved (1.8 points) in the TG versus control arm at T1 (p < 0.001). No adverse events were reported. The indoor TG seems safe and may reduce BPSD, medication intake, and cortisol levels in AD.


Opening Up: Clients' Inner Struggles in the Initial Phase of Therapy.
To explore how clients in clinical settings experience the process of opening up and sharing their inner experiences in the initial phase of therapy. Two psychotherapy sessions of clients (N = 11) were videotaped and followed by interviews. Interpersonal process recall was used to obtain in-depth descriptions of clients' immediate experiences in session. A follow-up interview was conducted 3 months later. The interviews were analyzed using thematic analysis. The data revealed how and why clients distanced themselves from inner experiences in the initial phase of therapy. The overarching theme was "Holding back and struggling to open up," which included four subthemes: (a) fearing the intensity and consequences of negative emotions; (b) experiences of being incapable and bodily stuck; (c) being insecure about one's worthiness and right to share inner experiences with the therapist; and (d) struggling with feeling disloyal to loved ones. The participants held back because they feared different consequences of opening up. A range of concerns led participants to distance themselves from their inner experiences and/or to refrain from openly talking about them to the therapist. Concerns related to appropriate interpersonal conduct as client were especially important. This knowledge is highly relevant to clinicians when building safety for psychotherapeutic work.


Genotype calling and haplotyping in parent-offspring trios.
Emerging sequencing technologies allow common and rare variants to be systematically assayed across the human genome in many individuals. In order to improve variant detection and genotype calling, raw sequence data are typically examined across many individuals. Here, we describe a method for genotype calling in settings where sequence data are available for unrelated individuals and parent-offspring trios and show that modeling trio information can greatly increase the accuracy of inferred genotypes and haplotypes, especially on low to modest depth sequencing data. Our method considers both linkage disequilibrium (LD) patterns and the constraints imposed by family structure when assigning individual genotypes and haplotypes. Using simulations, we show that trios provide higher genotype calling accuracy across the frequency spectrum, both overall and at hard-to-call heterozygous sites. In addition, trios provide greatly improved phasing accuracy--improving the accuracy of downstream analyses (such as genotype imputation) that rely on phased haplotypes. To further evaluate our approach, we analyzed data on the first 508 individuals sequenced by the SardiNIA sequencing project. Our results show that our method reduces the genotyping error rate by 50% compared with analysis using existing methods that ignore family structure. We anticipate our method will facilitate genotype calling and haplotype inference for many ongoing sequencing projects.


Intracoronary imaging of coronary atherosclerosis: validation for diagnosis, prognosis and treatment.
While coronary atherosclerosis is a leading cause of mortality, evaluation of coronary lesions was previously limited to either indirect angiographic assessment of the lumen silhouette or post mortem investigations. Intracoronary (IC) imaging modalities have been developed that allow for visualization and characterization of coronary atheroma in living patients. Used alone or in combination, these modalities have enhanced our understanding of pathobiological mechanisms of atherosclerosis, identified factors responsible for disease progression, and documented the ability of various medications to reverse the processes of plaque growth and destabilization. These methodologies have established a link between in vivo plaque characteristics and subsequent coronary events, thereby improving individual risk stratification, paving the way for risk-tailored systemic therapies and raising the option for pre-emptive interventions. Moreover, IC imaging is increasingly used during coronary interventions to support therapeutic decision-making in angiographically inconclusive disease, guide and optimize procedural results in selected lesion and patient subsets, and unravel mechanisms underlying stent failure. This review aims to summarize current evidence regarding the role of IC imaging for diagnosis and risk stratification of coronary atherosclerosis, and to describe its clinical role for guiding percutaneous coronary interventions. Future perspectives for in-depth plaque characterization using novel techniques and multimodality imaging approaches are also discussed.


Electrical impedance tomography spectroscopy (EITS) for human head imaging.
Electrical impedance tomography (EIT) is a recently developed medical imaging method which has practical advantages for imaging brain function as it is inexpensive, rapid and portable. Its principal use in validated human studies to date has been to image changes in impedance at a single excitation frequency over time, but there are potential applications where it is desirable to obtain images from a single point in time, which could be achieved by imaging over multiple frequencies. We describe a novel multifrequency EIT design which provides up to 64 electrodes for imaging in the head. This was achieved by adding a multiplexer to a single channel of an existing system, the Sheffield Mark 3.5. This provides a flexible protocol for addressing up to 64 electrodes but CMRR decreases from 90 dB to 80 dB and analogue amplifier bandwidth from > 1.6 MHz to 0.8 MHz. This did not significantly affect performance, as cylinders of banana, 10% of the diameter of a saline filled spherical tank, could be visualized with frequency referenced imaging. The design appears to have been an acceptable compromise between practicality and performance and will now be employed in clinical trials of multifrequency EIT in stroke, epilepsy and neonatal brain injury.


Recruitment and follow-up of adolescent and young adult cancer survivors: the AYA HOPE Study.
Cancer is rare in adolescents and young adults (AYA), but these patients have seen little improvement in survival in contrast to most other age groups. Furthermore, participation in research by AYAs is typically low. We conducted a study to examine the feasibility of recruiting a population-based sample of AYA survivors to examine issues of treatment and health outcomes. Individuals diagnosed in 2007-08 and age 15-39 at the time of diagnosis with acute lymphocytic leukemia, Hodgkin lymphoma, non-Hodgkin lymphoma, germ cell cancer or sarcoma were identified by 7 Surveillance, Epidemiology, and End-Results (SEER) cancer registries, mailed surveys within 14 months after diagnosis and again a year later, and had medical records reviewed. 525 (43%) of the eligible patients responded, 39% refused and 17% were lost to follow-up. Extensive efforts were required for most potential respondents (87%). 76% of respondents completed the paper rather than online survey version. In a multivariate model, age, cancer site, education and months from diagnosis to the first mailing of the survey were not associated with participation, although males (p < 0.01), Hispanics and non-Hispanic blacks (p < 0.001) were less likely to participate. 91% of survivors completing the initial survey completed the subsequent survey. Despite the response rate, those who participated adequately reflected the population of AYA cancer survivors. The study demonstrates that cancer registries are valuable foundations for conducting observational, longitudinal population-based research on AYA cancer survivors. Achieving a reasonable response rate in this population is possible, but requires extensive resources.


Sharp upturn of life expectancy in the Netherlands: effect of more health care for the elderly?
During the 1980s and 1990s life expectancy at birth has risen only slowly in the Netherlands. In 2002, however, the rise in life expectancy suddenly accelerated. We studied the possible causes of this remarkable development. Mortality data by age, gender and cause of death were analyzed using life table methods and age-period-cohort modeling. Trends in determinants of mortality (including health care delivery) were compared with trends in mortality. Two-thirds of the increase in life expectancy at birth since 2002 were due to declines in mortality among those aged 65 and over. Declines in mortality reflected a period rather than a cohort effect, and were seen for a wide range of causes of death. Favorable changes in mortality determinants coinciding with the acceleration of mortality decline were mainly seen within the health care system. Health care expenditure rose rapidly after 2001, and was accompanied by a sharp rise of specialist visits, drug prescriptions, hospital admissions and surgical procedures among the elderly. A decline of deaths following non-treatment decisions suggests a change towards more active treatment of elderly patients. Our findings are consistent with the idea that the sharp upturn of life expectancy in the Netherlands was at least partly due to a sharp increase in health care for the elderly, and has been facilitated by a relaxation of budgetary constraints in the health care system.


Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors.
Cytomegalovirus (CMV) elicits long-term T-cell immunity of unparalleled strength, which has allowed the development of highly protective CMV-based vaccine vectors. Counterintuitively, experimental vaccines encoding a single MHC-I restricted epitope offered better immune protection than those expressing entire proteins, including the same epitope. To clarify this conundrum, we generated recombinant murine CMVs (MCMVs) encoding well-characterized MHC-I epitopes at different positions within viral genes and observed strong immune responses and protection against viruses and tumor growth when the epitopes were expressed at the protein C-terminus. We used the M45-encoded conventional epitope HGIRNASFI to dissect this phenomenon at the molecular level. A recombinant MCMV expressing HGIRNASFI on the C-terminus of M45, in contrast to wild-type MCMV, enabled peptide processing by the constitutive proteasome, direct antigen presentation, and an inflation of antigen-specific effector memory cells. Consequently, our results indicate that constitutive proteasome processing of antigenic epitopes in latently infected cells is required for robust inflationary responses. This insight allows utilizing the epitope positioning in the design of CMV-based vectors as a novel strategy for enhancing their efficacy.


Vestibular perception and the vestibulo-ocular reflex in young and older adults.
Quantification of the perceptual thresholds to vestibular stimuli may offer valuable complementary information to that provided by measures of the vestibulo-ocular reflex (VOR). Perceptual thresholds could be particularly important in evaluating some subjects, such as the elderly, who might have a greater potential of central as well as peripheral vestibular dysfunction. The authors hypothesized that perceptual detection and discrimination thresholds would worsen with aging, and that there would be a poor relation between thresholds and traditional measures of the angular VOR represented by gain and phase on rotational chair testing. The authors compared the detection and discrimination thresholds of 19 younger and 16 older adults in response to earth-vertical, 0.5 Hz rotations. Perceptual results of the older subjects were then compared with the gain and phase of their VOR in response to earth-vertical rotations over the frequency range from 0.025 to 0.5 Hz. Detection thresholds were found to be 0.69 ± 0.29 degree/sec (mean ± standard deviation) for the younger participants and 0.81 ± 0.42 degree/sec for older participants. Discrimination thresholds in younger and older adults were 4.83 ± 1.80 degree/sec and 4.33 ± 1.57 degree/sec, respectively. There was no difference in either measure between age groups. Perceptual thresholds were independent of the gain and phase of the VOR. These results indicate that there is no inevitable loss of vestibular perception with aging. Elevated thresholds among the elderly are therefore suggestive of pathology rather than normal consequences of aging. Furthermore, perceptual thresholds offer additional insight, beyond that supplied by the VOR alone, into vestibular function.


Penetration of efavirenz into the male genital tract: drug concentrations and antiviral activity in semen and blood of HIV-1-infected men.
Efavirenz is a potent non-nucleoside reverse transcriptase inhibitor, licensed for the treatment of HIV-1. Data on sanctuary site penetration are limited. Therefore, we measured efavirenz concentrations in the blood and semen of 19 HIV-1-positive men and found concentrations in seminal plasma averaged 10% of those in blood plasma. Furthermore, seminal plasma viral loads were suppressed by 24 weeks of therapy in all patients. These data suggest that efavirenz-containing regimens have antiviral activity within the male genital tract.


Core values versus common sense: consequentialist views appear less rooted in morality.
When a speaker presents an opinion, an important factor in audiences' reactions is whether the speaker seems to be basing his or her decision on ethical (as opposed to more pragmatic) concerns. We argue that, despite a consequentialist philosophical tradition that views utilitarian consequences as the basis for moral reasoning, lay perceivers think that speakers using arguments based on consequences do not construe the issue as a moral one. Five experiments show that, for both political views (including real State of the Union quotations) and organizational policies, consequentialist views are seen to express less moralization than deontological views, and even sometimes than views presented with no explicit justification. We also demonstrate that perceived moralization in turn affects speakers' perceived commitment to the issue and authenticity. These findings shed light on lay conceptions of morality and have practical implications for people considering how to express moral opinions publicly.


Infection as Trigger for Congenital Thrombotic Thrombocytopenic Purpura in an Adult Patient.
Congenital thrombotic thrombocytopenic purpura (cTTP) is an inherited disease that is sometimes fatal in early childhood. cTTP is similar to idiopathic thrombotic thrombocytopenic purpura (iTTP); both are characterized by varying levels of thrombocytopenia, microangiopathic hemolytic anemia (MAHA), and end-organ damage secondary to occlusion of the microvasculature. cTTP is caused by a partial or total deficiency or loss of function of ADAMTS-13 (a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13). We report the case of a 33-year-old woman who was mistakenly diagnosed with primary immune thrombocytopenia (ITP) during childhood. The patient was referred to our center with dyspnea, fatigue, fever, and jaundice with no clinical bleeding. Laboratory features were compatible with MAHA; ADAMTS-13 activity was at 0%, with negativity for ADAMTS-13 antibodies. We concluded the final diagnosis was cTTP. The triggering factor identified for MAHA was a double infection: central venous catheter bacterial infection and atypical pneumonia. After 7 days of treatment with antibiotics and ongoing total plasma exchange (TPE), the patient responded favorably. Our patient received fresh frozen plasma (FFP) infusion once every 2 weeks, and prophylactic voriconazole remained under control at the time of writing. As demonstrated in this case, effective treatment of the trigger cause helps reduce the need for continuous FFP exposure and controls the MAHA.


Expression of the Listeria monocytogenes EGD inlA and inlB genes, whose products mediate bacterial entry into tissue culture cell lines, by PrfA-dependent and -independent mechanisms.
Internalization of Listeria monocytogenes into nonphagocytic cell lines in vitro requires the products of the inlAB locus (J.-L. Gaillard, P. Berche, C. Frehel, E. Gouin, and P. Cossart, Cell 65:1127-1141, 1991). By generating isogenic mutants with a chromosomal in-frame deletion in either inlA or inlB, we have identified InlA and InlB as surface-bound proteins of L. monocytogenes with molecular weights of 88,000 and 65,000, respectively. These results were obtained with monoclonal antibodies raised against either protein and corroborated by N-terminal end sequencing of InlA and InlB. By immunoblot analysis, the production of both polypeptides was found to be strongly dependent on growth temperature and, particularly for InlB, on the presence of the PrfA regulator protein. Expression of InlA was not strictly dependent on the presence of the PrfA regulator protein. Transcription analysis of the inlAB locus revealed that the inlA gene was transcribed by several promoters, of which only one is PrfA dependent. This PrfA-dependent inlA promoter, which contains two base substitutions within its putative PrfA DNA-binding palindrome, is responsible for transcription of both inlA and inlB genes. A hitherto unrecognized promoter located 51 bp upstream of the GTG start codon of the inlB gene was also detected. Hence, inlA and inlB are transcribed both individually and in an operon by PrfA-dependent and -independent mechanisms. Tissue culture invasion assays employing various epithelial cell lines demonstrated that both InlA and InlB are required for invasion. In vivo studies using the mouse infection model revealed that both internalin mutants were attenuated for virulence.


The impact of COVID-19 pandemic on vascular registries and clinical trials.
Quality improvement programs and clinical trial research experienced disruption due to the coronavirus disease 2019 (COVID-19) pandemic. Vascular registries showed an immediate impact with significant declines in second-quarter vascular procedure volumes witnessed across Europe and the United States. To better understand the magnitude and impact of the pandemic, organizations and study groups sent grass roots surveys to vascular specialists for needs assessment. Several vascular registries responded quickly by insertion of COVID-19 variables into their data collection forms. More than 80% of clinical trials have been reported delayed or not started due to factors that included loss of enrollment from patient concerns or mandated institutional shutdowns, weighing the risk of trial participation on patient safety. Preliminary data of patients undergoing vascular surgery with active COVID-19 infection show inferior outcomes (morbidity) and increased mortality. Disease-specific vascular surgery study collaboratives about COVID-19 were created for the desire to study the disease in a more focused manner than possible through registry outcomes. This review describes the pandemic effect on multiple VASCUNET registries including Germany (GermanVasc), Sweden (SwedVasc), United Kingdom (UK National Vascular Registry), Australia and New Zealand (bi-national Australasian Vascular Audit), as well as the United States (Society for Vascular Surgery Vascular Quality Initiative). We will highlight the continued collaboration of VASCUNET with the Vascular Quality Initiative in the International Consortium of Vascular Registries as part of the Medical Device Epidemiology Network coordinated registry network. Vascular registries must remain flexible and responsive to new and future real-world problems affecting vascular patients.


The genomic landscape of balanced cytogenetic abnormalities associated with human congenital anomalies.
Despite the clinical significance of balanced chromosomal abnormalities (BCAs), their characterization has largely been restricted to cytogenetic resolution. We explored the landscape of BCAs at nucleotide resolution in 273 subjects with a spectrum of congenital anomalies. Whole-genome sequencing revised 93% of karyotypes and demonstrated complexity that was cryptic to karyotyping in 21% of BCAs, highlighting the limitations of conventional cytogenetic approaches. At least 33.9% of BCAs resulted in gene disruption that likely contributed to the developmental phenotype, 5.2% were associated with pathogenic genomic imbalances, and 7.3% disrupted topologically associated domains (TADs) encompassing known syndromic loci. Remarkably, BCA breakpoints in eight subjects altered a single TAD encompassing MEF2C, a known driver of 5q14.3 microdeletion syndrome, resulting in decreased MEF2C expression. We propose that sequence-level resolution dramatically improves prediction of clinical outcomes for balanced rearrangements and provides insight into new pathogenic mechanisms, such as altered regulation due to changes in chromosome topology.


Crystal structure of the Haemophilus influenzae Hap adhesin reveals an intercellular oligomerization mechanism for bacterial aggregation.
Bacterial biofilms are complex microbial communities that are common in nature and are being recognized increasingly as an important determinant of bacterial virulence. However, the structural determinants of bacterial aggregation and eventual biofilm formation have been poorly defined. In Gram-negative bacteria, a major subgroup of extracellular proteins called self-associating autotransporters (SAATs) can mediate cell-cell adhesion and facilitate biofilm formation. In this study, we used the Haemophilus influenzae Hap autotransporter as a prototype SAAT to understand how bacteria associate with each other. The crystal structure of the H. influenzae Hap(S) passenger domain (harbouring the SAAT domain) was determined to 2.2 Å by X-ray crystallography, revealing an unprecedented intercellular oligomerization mechanism for cell-cell interaction. The C-terminal SAAT domain folds into a triangular-prism-like structure that can mediate Hap-Hap dimerization and higher degrees of multimerization through its F1-F2 edge and F2 face. The intercellular multimerization can give rise to massive buried surfaces that are required for overcoming the repulsive force between cells, leading to bacterial cell-cell interaction and formation of complex microcolonies.


Antibiotic resistance as collateral damage: the tragedy of the commons in a two-disease setting.
We propose a simple two-disease epidemic model where one disease exhibits only a drug-sensitive strain, while the other exhibits both drug-sensitive and drug-resistant strains. Treatment for the first disease may select for resistance in the other. We model antibiotic use as a mathematical game through the study of individual incentives and community welfare. The basic reproduction number is derived and the existence and local stability of the model equilibria are analyzed. When the force of infection of each disease is unaffected by the presence of the other, we find that there is a conflict of interest between individual and community, known as a tragedy of the commons, under targeted treatment toward persons infected by the single strain disease, but there is no conflict under mass treatment. However, we numerically show that individual and social incentive to use antibiotics may show disaccord under mass treatment if the restriction on the transmission ability of the dually infected people is removed, or drug resistant infection is worse than drug sensitive infection, or the uninfected state has a comparative disutility over the infected states.


Development of Small Molecule MEIS Inhibitors that modulate HSC activity.
Meis1, which belongs to TALE-type class of homeobox gene family, appeared as one of the key regulators of hematopoietic stem cell (HSC) self-renewal and a potential therapeutical target. However, small molecule inhibitors of MEIS1 remained unknown. This led us to develop inhibitors of MEIS1 that could modulate HSC activity. To this end, we have established a library of relevant homeobox family inhibitors and developed a high-throughput in silico screening strategy against homeodomain of MEIS proteins using the AutoDock Vina and PaDEL-ADV platform. We have screened over a million druggable small molecules in silico and selected putative MEIS inhibitors (MEISi) with no predicted cytotoxicity or cardiotoxicity. This was followed by in vitro validation of putative MEIS inhibitors using MEIS dependent luciferase reporter assays and analysis in the ex vivo HSC assays. We have shown that small molecules named MEISi-1 and MEISi-2 significantly inhibit MEIS-luciferase reporters in vitro and induce murine (LSKCD34l°w cells) and human (CD34+, CD133+, and ALDHhi cells) HSC self-renewal ex vivo. In addition, inhibition of MEIS proteins results in downregulation of Meis1 and MEIS1 target gene expression including Hif-1α, Hif-2α and HSC quiescence modulators. MEIS inhibitors are effective in vivo as evident by induced HSC content in the murine bone marrow and downregulation of expression of MEIS target genes. These studies warrant identification of first-in-class MEIS inhibitors as potential pharmaceuticals to be utilized in modulation of HSC activity and bone marrow transplantation studies.


Analysis and interpretation of a unique Arabic finger ring from the Viking Age town of Birka, Sweden.
In this work we used non-destructive SEM imaging and EDS analysis to characterize the material composition of an Arabic finger ring, which was found in a 9(th) c. woman's grave at the Viking Age (A.D. 793-1066) trading center of Birka, Sweden. The ring is set with a violet stone inscribed with Arabic Kufic writing, here interpreted as reading "il-la-lah", i.e. "For/to Allah". The stone was previously thought to be an amethyst, but the current results show it to be coloured glass. The ring has been cast in a high-grade silver alloy (94.5/5.5 Ag/Cu) and retains the post-casting marks from the filing done to remove flash and mold lines. Thus, the ring has rarely been worn, and likely passed from the silversmith to the woman buried at Birka with few owners in between. The ring may therefore constitute material evidence for direct interactions between Viking Age Scandinavia and the Islamic world. Being the only ring with an Arabic inscription found at a Scandinavian archaeological site, it is a unique object among Swedish Viking Age material. The technical analysis presented here provides a better understanding of the properties and background of this intriguing piece of jewelry.


Cigarette smoke-induced induction of antioxidant enzyme activities in airway leukocytes is absent in active smokers with COPD.
Oxidative injury to the airway has been proposed as an important underlying mechanism in the pathogenesis of chronic obstructive pulmonary disease (COPD). As the extent of oxidant-mediated damage is dependent on the endogenous antioxidant defences within the airways, we examined whether COPD was associated with deficiencies in the antioxidant network within the respiratory tract lining fluids (RTLFs) and resident airway leukocytes. We hypothesised that COPD would be associated with both basal depression of antioxidant defences and impaired adaptive antioxidant responses to cigarette smoke. Low molecular weight and enzymatic antioxidants together with metal-handling proteins were quantified in bronchoalveolar lavage fluid and airway leukocytes, derived from current (n=9) and ex-smoking COPD patients (n=15), as well as from smokers with normal lung function (n=16) and healthy never smokers (n=13). Current cigarette smoking was associated with an increase in ascorbate and glutathione within peripheral RTLFs in both smokers with normal lung function compared with healthy never smokers and in COPD smokers compared with COPD ex-smokers. In contrast, intra-cellular antioxidant enzyme activities (glutathione peroxidase, glutathione reductase, and catalase) were only up-regulated in smokers with normal lung function compared with healthy never smokers and not in actively smoking COPD patients relative to COPD ex-smokers. We found no evidence of impaired basal antioxidant defences, within either the RTLFs or airway leukocytes in stable ex-smoking COPD patients compared with healthy never smoking controls. Current cigarette smoking induced an up-regulation of low molecular weight antioxidants in the RTLFs of both control subjects with normal lung function and patients with COPD. Importantly, the present data demonstrated a cigarette smoke-induced increase in intra-cellular antioxidant enzyme activities only within the smokers with normal lung function, implying that patients with COPD who continue to smoke will experience enhanced oxidative stress, prompting disease progression.


Data mining the human gut microbiota for therapeutic targets.
It is well known that microbes have an intricate role in human health and disease. However, targeted strategies for modulating human health through the modification of either human-associated microbial communities or associated human-host targets have yet to be realized. New knowledge about the role of microbial communities in the microbiota of the gastrointestinal tract (GIT) and their collective genomes, the GIT microbiome, in chronic diseases opens new opportunities for therapeutic interventions. GIT microbiota participation in drug metabolism is a further pharmaceutical consideration. In this review, we discuss how computational methods could lead to a systems-level understanding of the global physiology of the host-microbiota superorganism in health and disease. Such knowledge will provide a platform for the identification and development of new therapeutic strategies for chronic diseases possibly involving microbial as well as human-host targets that improve upon existing probiotics, prebiotics or antibiotics. In addition, integrative bioinformatics analysis will further our understanding of the microbial biotransformation of exogenous compounds or xenobiotics, which could lead to safer and more efficacious drugs.


Activation of knot (kn) specifies the 3-4 intervein region in the Drosophila wing.
Hedgehog (Hh) plays an important role in Drosophila wing patterning by inducing expression of Dpp, which serves to organize the wing globally across the A-P axis. We show here how Hh signalling also plays a direct role in patterning the medial wing through the activation of the Hh-target gene, knot (kn). kn is expressed in Hh-responsive cells near the A-P compartment boundary, where its expression is dependent on fu, a component of Hh signalling. kn is required for the proper positioning of veins 3 and 4 and to prevent ectopic venation between them. Furthermore, the expansion anteriorly of the normal kn expression domain causes an associated anterior shift in the position of vein 3 in the resultant wing. Ectopic expression of kn elsewhere in the wing imaginal disc results in the failure to properly activate the vein initiation genes, rho and Dl. Expression of the gene encoding the EGF-receptor (EGFR), which is required for vein initiation and subsequent differentiation, is normally depressed in the 3-4 intervein region. This downregulation of EGFR in the medial portion of the imaginal disc is dependent on kn activity and ectopic expression of kn inactivates EGFR elsewhere in the wing primordium. We propose kn expression in Hh-responsive cells of the wing blade anlagen during the late third instar creates a zone of cells in the medial wing in which vein primordia cannot be induced. The primordia for veins 3 and 4 are laid down adjacent to the kn-imposed vein-free zone, presumably by a signalling factor (such as Vn) also synthesized in the medial region of the wing.


Longitudinal evaluation of the impact of placement development teams on student support in clinical practice.
To investigate the impact of a new structure for supporting healthcare students and mentors in practice placements (Placement Development Teams). The English Model National Partnership Agreement for healthcare education required Strategic Health Authorities, Higher Education Institutions and National Health Service Trusts to redesign strategies for student support. Placement Development Teams are one English University's response to this. This study was phase 2 of a longitudinal qualitative evaluation of Placement Development Teams. Data were collected after establishment of Placement Development Teams, and compared and contrasted with those collected prior to their implementation. Telephone interviews were conducted with key educational stakeholders in Trusts and Strategic Health Authorities. Focus groups were conducted with third year non-medical healthcare students and first year paramedics working in 16 NHS Trusts in the south west peninsula of England. Pre-Placement Development Teams, themes from the students' data were: Supportive and unsupportive behaviour of staff; Mentor allocation; Placement allocation; Benefits of students to the placement area and Perceived control over the learning experience. Post-Placement Development Teams, the themes were Communication; Supportive and unsupportive behaviour of staff; The effect of peers on the placement experience; Knowledge and perceptions of the work of the PDTs. Form the staff data, pre-Placement Development Teams the themes were: Vision for improving student support. Post-Placement Development Teams themes from the staff data were how they provided a central point of contact for student and mentor support; and how they supported students and mentors. Support of students and mentors is particularly important following the introduction of The English Model National Partnership Agreement for healthcare education. Placement Development Teams can facilitate partnership working between higher education institutions and placement providers for student support.


Diffusion rates of cell surface antigens of mouse-human heterokaryons. III. Regulation of lateral diffusion rates by calcium ions.
In mouse-human heterokaryons, the lateral diffusion of major histocompatibility (MHC) antigens in the plasma membrane is enhanced by treatment of parent cells with ouabain. Ouabain treatment is ineffective if the medium lacks calcium ion, or if Verapamil, a blocker of calcium channels, is present. The divalent ionophore A23187 also enhances lateral diffusion of MHC antigens, to the same extent as ouabain, A23187 is effective only if calcium is present in the medium. Thus it appears that increased levels of cell calcium release constraints to lateral diffusion of MHC antigens.


Crystal structure of human myo-inositol monophosphatase 2, the product of the putative susceptibility gene for bipolar disorder, schizophrenia, and febrile seizures.
The human IMPA2 gene, which encodes myo-inositol monophosphatase 2 (IMPA2), is mapped onto 18p11.2, a susceptibility region for bipolar disorder. This chromosomal region has also been proposed to include a susceptibility locus for schizophrenia and febrile seizures. Here we report the crystal structures of human IMPA2 and its complex with calcium and phosphate ions. Human IMPA2 comprises an alpha-beta protein with a five-layered sandwich of alpha-helices and beta-sheets (alpha-beta-alpha-beta-alpha). The crystal structure and analytical ultracentrifugation results indicated that IMPA2 exists as a dimer in solution. The overall structure of IMPA2 is similar to that of IMPA1, except for the loop regions. In IMPA1, the loop region (31-43) is located at the entrance of the active site cavity. In the corresponding region (42-54) of IMPA2, the residues are disordered and partially form an alpha-helix. The structural difference in the opening of the active site cavity suggests that the substrate specificity differs between IMPA1 and IMPA2. The widely opened cavity of IMPA2 implies that the physiological substrate may be a larger compound than inositol monophosphate. The structure of IMPA2 complexed with Ca2+ revealed two metals and one phosphate binding sites, which were the same sites as in IMPA1 complexed with Mn2+ and phosphate, suggesting that the mechanism of the enzymatic reaction is similar to that of IMPA1. The crystal structures of human IMPA2 are useful for understanding the effect of nonsynonymous polymorphism reported in IMPA2, and will contribute to further functional analyses of IMPA2 that potentially predisposes to the vulnerabilities of bipolar disorder, schizophrenia, and febrile seizures.


Functional plasticity in oyster gut microbiomes along a eutrophication gradient in an urbanized estuary.
Oysters in coastal environments are subject to fluctuating environmental conditions that may impact the ecosystem services they provide. Oyster-associated microbiomes are responsible for some of these services, particularly nutrient cycling in benthic habitats. The effects of climate change on host-associated microbiome composition are well-known, but functional changes and how they may impact host physiology and ecosystem functioning are poorly characterized. We investigated how environmental parameters affect oyster-associated microbial community structure and function along a trophic gradient in Narragansett Bay, Rhode Island, USA. Adult eastern oyster, Crassostrea virginica, gut and seawater samples were collected at 5 sites along this estuarine nutrient gradient in August 2017. Samples were analyzed by 16S rRNA gene sequencing to characterize bacterial community structures and metatranscriptomes were sequenced to determine oyster gut microbiome responses to local environments. There were significant differences in bacterial community structure between the eastern oyster gut and water samples, suggesting selection of certain taxa by the oyster host. Increasing salinity, pH, and dissolved oxygen, and decreasing nitrate, nitrite and phosphate concentrations were observed along the North to South gradient. Transcriptionally active bacterial taxa were similar for the different sites, but expression of oyster-associated microbial genes involved in nutrient (nitrogen and phosphorus) cycling varied throughout the Bay, reflecting the local nutrient regimes and prevailing environmental conditions. The observed shifts in microbial community composition and function inform how estuarine conditions affect host-associated microbiomes and their ecosystem services. As the effects of estuarine acidification are expected to increase due to the combined effects of eutrophication, coastal pollution, and climate change, it is important to determine relationships between host health, microbial community structure, and environmental conditions in benthic communities.


Behavior mapping: a method for linking preschool physical activity and outdoor design.
The preschool that children attend has been shown to be a significant but variable predictor of physical activity of 3- to 5-yr-olds, whereas the time outdoors has been found to be a strong correlate of physical activity. Researchers speculate that variations in preschool physical activity may be attributed to variations in preschool policies and practices, including the form and content of outdoor physical environments. However, assessment methods linking them to physical activity are limited. Improved understanding of links between environment and activity could be used to influence childcare policy, which is highly regulated, usually at state level, to create outdoor environments more conducive to children's informal play and physical activity. The purpose of this article was to introduce behavior mapping as a direct observation method on the basis of the theories of behavior setting and affordance and to demonstrate its sensitivity to gathering physical activity and associated environmental data at a sufficiently detailed level to affect built environment design policy. Behavior mapping data, including outdoor environmental characteristics and children's physical activity levels, were obtained in two preschool centers at the Research Triangle region, NC. Physical activity levels at the two centers varied across different types of behavior settings, including pathways, play structures, and open areas. The same type of setting with different attributes, such as circular versus straight pathways, and open areas with different ground surfaces, such as asphalt, compacted soil, woodchips, and sand, attracted different levels of physical activity. Behavior mapping provides a promising method for objectively measuring relationships between physical behavior settings and directly associated activity levels.


Building bridges in dietary counselling: an exploratory study examining the usefulness of wellness and wellbeing concepts.
Over the last decade, professional discourse around health ownership has been evolving to recognise an individually-driven wellness/wellbeing approach. Concurrently, dietetic competencies have changed to include client-centred counselling incorporating individual client's perspectives within dietary prescriptions. The present exploratory research aimed to explore how client-centred counselling practice was being represented in the dietetic literature and to examine dietitians' perspectives about working with clients in the current environment. To explore the professional position, a literature search was conducted using keywords encompassing client-centred care and competency within professional dietetic journals (2001-2010). To develop a contextual case study, 10 in-depth interviews with dietitians delivering weight-loss prescriptions within a clinical trial were conducted. Recordings of their perspectives on roles, opportunities/barriers and counselling strategies were transcribed verbatim and examined using inductive thematic and content analysis. Eleven articles were incorporated into a narrative review describing practice issues related to traditional forms of consultation and the effectiveness of client-centred approaches. The over-riding theme from the interviews (Professional Identity Dilemma) highlighted tension felt by dietitians in their dual role as nutrition expert and counsellor, trained to deliver biomedical imperatives (clinical targets), and their challenge to accept client-defined health perspectives. Supporting themes (Adherence factors and Constructs of health) exposed details on barriers to dietary change and the impact of contextual factors on this change that were linked to wellness and wellbeing concepts. Appreciating wellness and wellbeing concepts may add a useful adjunct to client-centred approaches to dietary counselling through building bridges between clinical targets and client health perspectives.


International medical electives in selected African countries: a phenomenological study on host experience.
To explore the host experience on international medical electives at a selection of hospitals in low- and middle-income countries in Africa. Outcomes of the study may inform and improve the preparation of global health curriculum, pre-elective training and debriefing for international medical electives. A descriptive phenomenological study was undertaken, involving semi-structured interviews with ten elective hosts at seven study sites in three African countries. Purposive convenience sampling augmented by snowballing was utilised to recruit study participants. The data were thematically analysed and interpreted with reflexivity to generate an accurate aggregate of the experience of participants in hosting international medical electives. Six main themes emerged from the thematic analysis of interview data: international medical student contribution to host hospitals, host professional and personal fulfiment, barriers to student learning experience, international medical student preparedness, hope for reciprocity and barriers to cultural immersion and patient care. Study participants described the experience of hosting international medical elective students as overwhelmingly positive. However, they highlighted issues such as barriers to students' learning experience and the lack of reciprocity between host and sending institutions as areas which could be addressed to optimize the experience for both hosts and international medical students. An understanding of the host experience provides stakeholders with a clearer idea of what is important in preparation, organisation and evaluation of the elective experience. This study provides the impetus for further research to examine the effectiveness of introducing appropriate pre-departure training and post-elective debriefing to students embarking on international medical electives.


The design and statistical aspects of VIETNARMS: a strategic post-licensing trial of multiple oral direct-acting antiviral hepatitis C treatment strategies in Vietnam.
Eliminating hepatitis C is hampered by the costs of direct-acting antiviral treatment and the need to treat hard-to-reach populations. Access could be widened by shortening or simplifying treatment, but limited research means it is unclear which approaches could achieve sufficiently high cure rates to be acceptable. We present the statistical aspects of a multi-arm trial designed to test multiple strategies simultaneously and a monitoring mechanism to detect and stop individual randomly assigned groups with unacceptably low cure rates quickly. The VIETNARMS trial will factorially randomly assign patients to two drug regimens, three treatment-shortening strategies or control, and adjunctive ribavirin or no adjunctive ribavirin with shortening strategies (14 randomly assigned groups). We will use Bayesian monitoring at interim analyses to detect and stop recruitment into unsuccessful strategies, defined by more than 0.95 posterior probability that the true cure rate is less than 90% for the individual randomly assigned group (non-comparative). Final comparisons will be non-inferiority for regimens (margin 5%) and strategies (margin 10%) and superiority for adjunctive ribavirin. Here, we tested the operating characteristics of the stopping guideline for individual randomly assigned groups, planned interim analysis timings and explored power at the final analysis. A beta (4.5, 0.5) prior for the true cure rate produces less than 0.05 probability of incorrectly stopping an individual randomly assigned group with a true cure rate of more than 90%. Groups with very low cure rates (<60%) are very likely (>0.9 probability) to stop after about 25% of patients are recruited. Groups with moderately low cure rates (80%) are likely to stop (0.7 probability) before overall recruitment finishes. Interim analyses 7, 10, 13 and 18 months after recruitment commences provide good probabilities of stopping inferior individual randomly assigned groups. For an overall true cure rate of 95%, power is more than 90% to confirm non-inferiority in the regimen and strategy comparisons, regardless of the control cure rate, and to detect a 5% absolute difference in the ribavirin comparison. The operating characteristics of the stopping guideline are appropriate, and interim analyses can be timed to detect individual randomly assigned groups that are highly likely to have suboptimal performance at various stages. Therefore, our design is suitable for evaluating treatment-shortening or -simplifying strategies. ISRCTN registry: ISRCTN61522291. Registered on 4 October 2019.


Protein oxidation in G0 cells of Saccharomyces cerevisiae depends on the state rather than rate of respiration and is enhanced in pos9 but not yap1 mutants.
Immunodetection of protein carbonyl groups demonstrates that growth arrest elicited by carbon or nitrogen starvation causes an increased oxidation of proteins in Saccharomyces cerevisiae. Mutant analysis suggests that the response regulator Pos9p is involved in mitigating self-inflicted oxidative damages in G(0) cells, whereas Yap1p is primarily required in growing cells. The data also suggest that oxidation of target proteins is not a priori an effect of arrest of cell division or nutrient depletion and cannot be explained by the respiratory activity alone nor a high ratio of catabolic/anabolic activity in G(0) cells. Instead, we observed that starvation elicits a transition in the respiratory state (from phosphorylating to nonphosphorylating respiration) and that this transition is associated with a stepwise increase in protein oxidation. During carbon starvation, this transition and increase in oxidation occurs immediately as the carbon source is depleted, growth is arrested, and the respiratory rate falls drastically. In contrast, during nitrogen starvation and excess carbon the respiratory state transition and stepwise increase in protein oxidation are markedly delayed and occur long after the nitrogen source has been depleted and division and growth-arrested. Oxidation in G(0) cells could be enhanced by treating cells with low concentrations of antimycin A and attenuated with myxothiazol, indicating that protein oxidation is intimately linked to reactive oxygen species generated by semiquinones of the Q-cycle. Thus, the work presented suggests that the degree of coupling in the mitochondrial respiratory apparatus rather then the overall rate of respiration affects the degree of protein oxidation in nondividing yeast cells.


Light-induced selective deposition of metals on gold-tipped CdSe-seeded CdS nanorods.
We introduce a facile approach for the selective deposition of metals on Au-tipped CdSe-seeded CdS nanorods that exploits the transfer of electrons from CdS to the Au tips upon UV excitation. This light-induced deposition method was used for the deposition of Pd under mild conditions, which produced a Pd/Au alloyed tip while preserving the rest of the semiconductor nanoarchitecture. The highly site-selective deposition method was extended to the deposition of Fe, yielding monodispersed, structurally complex Au core/Fe(x)O(y) hollow shell-tipped semiconductor nanorods. These structurally well-defined rods were found to exhibit magnetic functionality. The synthetic strategies described in this work expand on the range of metals that can be deposited on heterostructured semiconductor nanorods, opening up new avenues for the hierarchical buildup of structural complexity and therefore multifunctionality in nanoparticles.