Pre- and postjunctional muscarinic receptors in canine bronchi.
Cholinergic receptors on canine bronchial smooth muscle (mediating excitation) or on the cholinergic nerve endings (modulating acetylcholine release) have not yet been characterized. In this study, the effects of agents selective for the various muscarinic receptors on mechanical and electrical responses in isolated segments of canine bronchial smooth muscle (3rd-6th order) were investigated. Carbachol (Cch), bethanechol (Bch), oxotremorine (Oxo), and McNeil A343 (McN) produced membrane depolarization and contractions with a ranked potency order of Oxo approximately Cch greater than Bch greater than McN. Oxo-induced contractions were antagonized by pirenzepine (Pir), AF-DX 116 (AF-DX), or hexahydrosiladifenidol (Hexa); Schild analysis yielded pA2 values of 6.61, 6.81, and 7.40, respectively. Contractions induced by field stimulation (FS) were antagonized by Pir, AF-DX, or Hexa with a potency order of Hexa greater than AF-DX approximately Pir. FS responses were potentiated by lower (i.e., M1-selective) concentrations of Pir, and antagonized by McN. The non-M1 agonists and antagonists had no such effects. These findings indicate that the postjunctional receptors are of the M3 subtype, and that the inhibitory receptors on the cholinergic nerve endings are of the M1 subtype.


Novel Lines of Research on the Environmental and Human Health Impacts of Nut Consumption.
Nuts have formed part of human diets throughout the ages. In recent decades, research has shown they are key foods in dietary patterns associated with lower chronic disease risk. The current state of climate change, however, has introduced an imperative to review the impact of dietary patterns on the environment with a shift to plant-based diets. Nuts emerge as a significant source of protein in plant-based diets and are a minimally processed and sustainable food. Research in this area is evolving to drive better production methods in varying climate conditions. Nevertheless, nut consumption remains an important contributor to human health. The mechanisms of action can be explained in terms of the nutrients they deliver. Studies of nut consumption have linked components such as monounsaturated fatty acids, plant omega-3 fatty acids, antioxidants, and plant sterols to improved lipoprotein profiles, lower blood pressure, and reduced cardiovascular disease risk. Preliminary research also indicates possible beneficial effects of nut consumption on reproductive health. In any case, the ultimate effects of foods on health are the results of multiple interactive factors, so where nuts fit within dietary patterns is a significant consideration for research translation. This has implications for research methodologies, including categorization within food groups and inclusion in Healthy Dietary Indices. The aim of this narrative review is to outline new focal points for investigation that examine the environmental and some novel human health impacts of nut consumption and discuss future directions for research.


Influence of calcium hydroxide on the post-treatment pain in Endodontics: A systematic review.
Pain of endodontic origin has been a major concern to the patients and the clinicians for many years. Post-operative pain is associated with inflammation in the periradicular tissues caused by irritants egressing from root canal during treatment. It has been suggested that calcium hydroxide intra-canal medicament has pain-preventive properties because of its anti-microbial or tissue altering effects. Some dispute this and reasoned that calcium hydroxide may initiate or increase pain by inducing or increasing inflammation. To evaluate the effectiveness of calcium hydroxide in reducing the post-treatment pain when used as an intra-canal medicament. THE FOLLOWING DATABASES WERE SEARCHED: PubMed CENTRAL (until July 2013), MEDLINE, and Cochrane Database of Systematic Reviews. Bibliographies of clinical studies and reviews identified in the electronic search were analyzed for studies published outside the electronically searched journals. The primary outcome measure was to evaluate the post-treatment pain reduction when calcium hydroxide is used as an intra-canal medicament in patients undergoing root canal therapy. The reviews found some clinical evidence that calcium hydroxide is not very effective in reducing post-treatment pain when it is used alone, but its effectiveness can be increased when used in combination with other medicaments like chlorhexidine and camphorated monochlorophenol (CMCP). Even though calcium hydroxide is one of the most widely used intra-canal medicament due to its anti-microbial properties, there is no clear evidence of its effect on the post-treatment pain after the chemo-mechanical root canal preparation.


Postoperative singultus: an osteopathic approach.
Singultus, or hiccups, is a common medical condition. Despite exponential leaps in medicine, the pathophysiologic cause remains poorly defined. Persistent singultus has been associated with conditions such as pulmonary embolism and myocardial infarction. Singultus is also a well-known postoperative complication. The criterion standard of care for patients with singultus involves ruling out lethal pathologic causes, attempting physical stimulation with Valsava maneuvers or drinking water, and, if no relief has been achieved, administering drugs to ease the symptoms. The authors report a case of a man whose postoperative singultus was successfully managed with osteopathic manipulative treatment. This approach addresses many of the possible underlying neuromechanical causes of the aberrant reflex with minimal potential for adverse effects. Physicians should consider osteopathic manipulative treatment in the care of patients with singultus.


Initial Experience with Novel Single-Use Disposable Ureteroscopy: A Prospective, Single Arm 90-Day Trial of the Axis Ureteroscope.
Novel digital disposable ureteroscopes are becoming more commonplace in endourological practices. We trialed the Axis™ single-use ureteroscope, which is distributed within the United States by Dornier MedTech. Our goal was to demonstrate clinical equivalence to our reusable platform and to evaluate the cost-effectiveness of a single-use digital ureteroscope. We conducted a prospective 90-day trial where all flexible ureteroscopic procedures were completed using the single-use ureteroscope. The cases were monitored for scope failure, scope deficiencies and surgeon satisfaction scores via an immediate postoperative REDCap® survey. We also performed a cost analysis between reusable scopes and the single-use ureteroscope. The cost of the reusable platform included the amortized initial purchase, maintenance and clean processing. Over a 90-day period, we performed 93 flexible ureteroscopy procedures with a single-use ureteroscope, of which 74 were completed using Axis. After controlling for operating room time and disposable items used, the utilization of Axis was associated with an average reduction of $140.31 per case. Extrapolating the per-case savings over an annual case volume, we estimate a total savings of $56,127.15. There were no disposable scope malfunctions. There were 4 (4.3%) cases converted to reusable scopes due to image quality (1), inadequate secondary deflection (2) and a tight ureter (1). Mean±SD score for image quality, mobility and ergonomics was 9.1±1.1, 8.9±1.1, 9.3±1.1, respectively. The 90-day complication rates were equal to our controls. We demonstrated that the Axis single-use digital ureteroscope is equivalent in function to our digital reusable scope and reduced the cost of flexible ureteroscopy.


Novel antibiotic-loaded particles conferring eradication of deep tissue bacterial reservoirs for the treatment of chronic urinary tract infection.
A significant proportion of urinary tract infection (UTI) patients experience recurrent episodes, due to deep tissue infection and treatment-resistant bacterial reservoirs. Direct bladder instillation of antibiotics has proved disappointing in treating UTI, likely due to the failure of infused antibiotics to penetrate the bladder epithelium and accumulate to high enough levels to kill intracellular bacteria. This work investigates the use of nitrofurantoin loaded poly(lactic-co-glycolic acid) (PLGA) particles to improve delivery to intracellular targets for the treatment of chronic UTI. Using electrohydrodynamic atomisation, we produced particles with an average diameter of 2.8 μm. In broth culture experiments, the biodegradable particles were effective against a number of UTI-relevant bacterial strains. Dye-loaded particles demonstrated that intracellular delivery was achieved in all cells in 2D cultures of a human bladder epithelial progenitor cell line in a dose-dependent manner, achieving far higher efficiency and concentration than equivalent quantities of free drug. Time-lapse video microscopy confirmed that delivery occurred within 30 min of administration, to 100% of cells. Moreover, the particles were able to deliver the drug to cells through multiple layers of a 3D human bladder organoid model causing minimal cell toxicity, displaying superior killing of bacterial reservoirs harboured within bladder cells compared with unencapsulated drug. The particles were also able to kill bacterial biofilms more effectively than the free drug. These results illustrate the potential for using antibiotic-loaded microparticles to effectively treat chronic UTIs. Such a delivery method could be extrapolated to other clinical indications where robust intracellular delivery is required, such as oncology and gene therapy.


Topography of sleep slow wave activity in children with attention-deficit/hyperactivity disorder.
Sleep slow wave activity (SWA, EEG power between 1 and 4.5 Hz) is a major characteristic of non-rapid eye movement (NREM) sleep, which seems to be critically involved in cortical plasticity. Studies using high-density electroencephalography (hd-EEG) showed that the topographical distribution of SWA mirrors cortical maturation, expressing a local maximum that is characteristic for a certain age range. We compared the sleep EEG of children with attention-deficit/hyperactivity disorder (ADHD) with healthy controls to explore differences in sleep SWA. All-night hd-EEG recordings (128 electrodes) were performed in a group of nine children diagnosed with ADHD and nine age- and sex-matched healthy controls. SWA topography was calculated and contrasted between the groups. We found a local increase of SWA in a cluster of six electrodes over central regions in children with ADHD compared to control children (+17% ± 6% SE, p < .01). This group difference was specific for the SWA range and stable across the night. Children with ADHD showed a less mature topographical SWA distribution in comparison to healthy children of the same age and sex. This neuromaturational delay in ADHD is in accordance with neuroimaging and behavioral studies. Thus, our study supports the use of sleep SWA topography as a reliable imaging tool for the study of cortical plasticity.


Differences in Morphologic and Hemodynamic Characteristics for "PHASES-Based" Intracranial Aneurysm Locations.
Several recent prospective studies have found that unruptured intracranial aneurysms at various anatomic locations have different propensities for future rupture. This study aims to uncover the lack of understanding regarding rupture-prone characteristics, such as morphology and hemodynamic factors, associated with different intracranial aneurysm location. We investigated the characteristics of 311 unruptured aneurysms at our center. Based on the PHASES study, we separated and compared morphologic and hemodynamic characteristics among 3 aneurysm location groups: 1) internal carotid artery; 2) middle cerebral artery; and 3) anterior communicating, posterior communicating, and posterior circulation arteries. A mixed model statistical analysis showed that size ratio, low wall shear stress area, and pressure loss coefficient were different between the intracranial aneurysm location groups. In addition, a pair-wise comparison showed that ICA aneurysms had lower size ratios, lower wall shear stress areas, and lower pressure loss coefficients compared with MCA aneurysms and compared with the group of anterior communicating, posterior communicating, and posterior circulation aneurysms. There were no statistical differences between MCA aneurysms and the group of anterior communicating, posterior communicating, and posterior circulation aneurysms for morphologic or hemodynamic characteristics. ICA aneurysms may be subjected to less rupture-prone morphologic and hemodynamic characteristics compared with other locations, which could explain the decreased rupture propensity of intracranial aneurysms at this location.


Live cell imaging in Drosophila melanogaster.
Although many of the techniques of live cell imaging in Drosophila melanogaster are also used by the greater community of cell biologists working on other model systems, studying living fly tissues presents unique difficulties with regard to keeping the cells alive, introducing fluorescent probes, and imaging through thick, hazy cytoplasm. This article outlines the major tissue types amenable to study by time-lapse cinematography and different methods for keeping the cells alive. It describes various imaging and associated techniques best suited to following changes in the distribution of fluorescently labeled molecules in real time in these tissues. Imaging, in general, is a rapidly developing discipline, and recent advances in imaging technology are able to greatly extend what can be achieved with live cell imaging of Drosophila tissues. As far as possible, this article includes the latest technical developments and discusses likely future developments in imaging methods that could have an impact on research using Drosophila.


Roles of estrogen receptor and p21(Waf1) in bortezomib-induced growth inhibition in human breast cancer cells.
Proteasome inhibitors such as bortezomib constitute novel therapeutic agents that are currently in clinical use and in clinical trials. In some neoplasms, cyclin-dependent kinase inhibitors (CKI) such as p21(WAF1) have been proposed as key targets of proteasome inhibitors. p21(WAF1) expression can be modulated by p53, a tumor suppressor, and especially in breast cancer cells, by estrogen receptor alpha (ERα), which is highly relevant to cancer growth. We investigated the effects of bortezomib using a panel of six cancer cell lines with variable status of ERα or p53 and found that bortezomib inhibited the growth of all cell lines in the same concentration range irrespective of the ERα expression or the mutational status of p53. Bortezomib treatment significantly enhanced p21(WAF1) protein levels in all cell lines but with different mechanisms according to ERα status. In ERα-positive cells, bortezomib treatment caused a strong increase in p21(WAF1) mRNA, whereas in ERα-negative cells it predominantly enhanced p21(WAF1) protein levels suggesting a posttranslational mechanism of p21(WAF1) regulation in the ERα-negative cells. Moreover, the antiproliferative activity of bortezomib was prevented by ERα silencing or p21(WAF1) knockdown in ERα-positive cells. Collectively, our results highlight the potential roles of ERα and p21(WAF1) in growth inhibition of cancer cells mediated by proteasome inhibitors, such as bortezomib.


SERS substrate based on the flexible hybrid of polydimethylsiloxane and silver colloid decorated with silver nanoparticles.
Various flexible SERS sensors have attracted widespread concern in performing the direct identification of the analytes adsorbed on arbitrary surfaces. Here, a sample method was proposed to integrate plasmonic nanoparticles into polydimethylsiloxane (PDMS) to fabricate flexible substrate for the decoration of silver nanoparticles (AgNPs). The flexible SERS sensor based on AgNPs/AgNPs-PDMS offers highly sensitive Raman detection with enhancement factor up to 8.3 × 109, which can be attributed to the integrative effects from both the increase of the light absorption of the embedded AgNPs in PDMS substrate and the EM enhancement from the adjacent top-top, bottom-bottom and top-bottom AgNPs. After undergoing the cyclic mechanical deformation, the SERS substrate still maintains high mechanical stability and stable SERS signals. However, upon stretching the flexible substrate, there was an amusing phenomenon that SERS signals can be highly increased, which results from that the reduction of lateral nanogaps between top and bottom of the PDMS boundary strengthens the trigger of the plasmon coupling as demonstrated by the simulated result. This result reveals that the tuning and the coupling of the electromagnetic fields can be effectively controlled by the macroscopic mechanical solicitation. That will have an important significance for practical applications in strain-dependent sensors and detectors.


Role of surgical thoracic sympathetic interruption in treatment of facial blushing: a systematic review.
This paper aims to review the evidence to support the effectiveness of sympathectomy as a treatment for facial blushing in terms of relief of facial blushing, patient satisfaction, recurrence of blushing, patients regretting treatment and its associated complications. A systematic search strategy was performed in Ovid-Medline, Embase, Cochrane library and NICE. Studies reporting outcomes of sympathetic interruption in the treatment of facial blushing were retrieved. Nine studies met the inclusion criteria with 1369 patients included in the final analysis. The age range of patients was 8 to 74 years (from 7 studies) with 56% females. Mean follow up was 21 months in 8 studies (range 6 to 30 months). The pooled proportion of patients who had good relief of facial blushing was 78.30% (95% C.I. 58.20% - 98.39%). Complete satisfaction was reported in 84.02% (95% C.I. 71.71% - 96.33%). Compensatory sweating and gustatory sweating were the commonest complications occurring in 74.18% (95% C.I. 58.10% - 90.26%) and 24.42% (95% C.I. 12.22% - 36.61%) respectively. The estimated proportion of patients regretting surgery was 6.79% (C.I 2.08% 11.50%). Sympathetic interruption at T2 or T2-3 ganglia appears to be an effective treatment for facial blushing. However, lack of randomized trials comparing sympathetic interruption with non-surgical methods of treatment and heterogeneity of included studies with respect to assessment of outcome measures preclude strong evidence and definitive recommendations.


Meanings of knowledge and identity in public health nursing in a time of transition: interpretations of public health nurses' narratives.
A changing healthcare system affects the professional identity of nursing groups. Public health nursing has experienced challenges in balancing the paternalistic expert ideology and the empowerment participation ideology. A strong professional identity can legitimate nursing, and possibly influence the quality of nursing work. Narrations from practice can illuminate the nurses' theoretical and practical knowledge and help illustrate their collective professional identity. To illuminate the meaning of public health nursing knowledge and professional identity in a continuously changing public health nursing practice. A qualitative interview study with a purposeful sample of 23 Norwegian public health nurses was carried out. Data were analysed using phenomenological hermeneutics, a descriptive method inspired by Ricoeur's philosophy of interpretation. Three themes emerged (i) Being a generalist: emphasising the need for generalised knowledge and using clinical judgement, (ii) Being one who empowers: focusing on resources and coping strategies, (iii) Being occupied with individual problem solving: focusing on individuals with special needs, using standardised techniques and protocols, and lacking specialised knowledge. Interpretation of the nurses' stories illuminated their need for generalised evidence-based knowledge, but also the importance of using sound clinical judgement in a diverse complex practice, where service users need encouragement, support and expert advice. Time pressures can limit the nurses' involvement. Many had an individual problem-focus more than a primary prevention focus, in contrast to governmental regulations stating that Norwegian public health nurses should focus on health promotion and primary prevention. Public health nurses have a broad generalised knowledge of their special target group giving them a 'specialist generalist' role. Clarification of this role, in relation to jurisdictional borders, can create a strong identity at a time when healthcare policy promotes economic values, professional neutrality and increased collaboration.


Genetic, Biochemical, and Biophysical Methods for Studying FeS Proteins and Their Assembly.
FeS clusters containing proteins are structurally and functionally diverse and present in most organisms. Our understanding of FeS cluster production and insertion into polypeptides has benefited from collaborative efforts between in vitro and in vivo studies. The former allows a detailed description of FeS-containing protein and a deep understanding of the molecular mechanisms catalyzing FeS cluster assembly. The second allows to include metabolic and environmental constraints within the analysis of FeS homeostasis. The interplay and the cross talk between the two approaches have been a key strategy to reach a multileveled integrated understanding of FeS cluster homeostasis. In this chapter, we describe the genetic and biochemical/biophysical strategies that were used in the field of FeS cluster biogenesis, with the aim of providing the reader with a critical view of both approaches. In addition to the description of classic tricks and a series of recommendations, we will also discuss models as well as spectroscopic techniques useful to characterize FeS clusters such as UV-visible, Mössbauer, electronic paramagnetic resonance, resonance Raman, circular dichroism, and nuclear magnetic resonance.


Inefficient and incorrect processing of the Ac transposase transcript in iae1 and wild-type Arabidopsis thaliana.
As part of the analysis of the Arabidopsis mutant iae1-1 (increased Ac excision), quantitative studies of the Ac transposase transcript were conducted. The primary transcript of Ac contains three small introns (introns 1-3; mean size 89 bp) and one larger intron (intron 4; 387 bp). We analysed the splicing of intron 3 and intron 4 in wild-type Arabidopsis and the iae1-1 mutant. Our results demonstrate that the splicing of Ac introns 3 and 4 is inefficient (splicing efficiencies 57 and 30% respectively) compared with that of an intron of an endogenous Arabidopsis gene (PHYB intron 1; splicing efficiency 90%). The poor splicing efficiency of Ac intron 4 was found to correlate with aberrant processing. Steady state levels of total Ac transcript were higher in the iae1-1 mutant than wild-type, but the same aberrant processing occurred. The inefficient processing of Ac in Arabidopsis prompted us to construct an Ac element lacking introns (Ac::cDNA) in an attempt to increase transposition frequencies. Autonomous activity of the Ac::cDNA element was undetectable in Arabidopsis, despite its ability to transpose at high frequency in response to a strong transposase source (35S::transposase) in trans, and the demonstrable autonomy of the same element in tobacco. A number of smaller transcripts were detected in Arabidopsis lines containing Ac::cDNA or Ac. Analysis of these smaller transcripts revealed a high frequency of premature polyadenylation in exon 2 and splicing of cryptic introns.


Optimizing the residency application process: insights from neurological surgery during the pandemic virtual application cycle.
In this article, the authors describe the impact of the COVID-19 virtual match cycle and discuss approaches to optimize future cycles through applicant and neurosurgical education leadership insights. Anonymous surveys of neurosurgery program leaders (program directors and program chairs), program administrators (PAs), and 2020-2021 neurosurgery residency match applicants were distributed by the SNS, in conjunction with the Association of Resident Administrators in Neurological Surgery and AANS Young Neurosurgeons Committee. Responses were received from 77 (67.0%) of 115 PAs, 119 (51.7%) of 230 program leaders, and 124 (44.3%) of 280 applicants representing geographically diverse regions. During the virtual application cycle relative to the previous year, programs received more Electronic Residency Application Service applications (mean 314.8 vs 285.3, p < 0.0001) and conducted more applicant interviews (mean 45.2 vs 39.9, p = 0.0003). More than 50% of applicants applied to > 80 programs; 60.3% received ≤ 20 interview invitations, and 9% received > 40 invitations. Overall, 65% of applicants completed ≤ 20 interviews, whereas 34.7% completed > 20 interviews. Program leaders described one 4-week home subinternship (93.3%) and two 4-week external subinternships (68.9%) as optimal neurosurgical exposure; 62.8% of program leaders found the standardized letter of recommendation template to be somewhat (47.5%) or significantly (15.3%) helpful. Applicants, PAs, and program leaders all strongly preferred a hybrid model of in-person and virtual interview options for future application cycles over all in-person or all virtual options. Ninety-three percent of applicants reported matching within their top 10-ranked programs, and 52.9% of programs matched residents within the same decile ranking as in previous years. Optimizing a national strategy for the neurosurgery application process that prioritizes equity and reduces costs, while ensuring adequate exposure for applicants to gain educational opportunities and evaluate programs, is critical to maintain a successful training system.


The Rheb-TORC1 signaling axis functions as a developmental checkpoint.
In many eukaryotes, the small GTPase Rheb functions as a switch to toggle activity of TOR complex 1 (TORC1) between anabolism and catabolism, thus controlling lifespan, development and autophagy. Our CRISPR-generated, fluorescently tagged endogenous Caenorhabditis elegans RHEB-1 and DAF-15/Raptor are expressed ubiquitously and localize to lysosomes. LET-363/TOR and DAF-15/Raptor are required for development beyond the third larval stage (L3). We observed that deletion of RHEB-1 similarly conferred L3 arrest. Unexpectedly, robust RNAi-mediated depletion of TORC1 components caused arrest at stages prior to L3. Accordingly, conditional depletion of endogenous DAF-15/Raptor in the soma revealed that TORC1 is required at each stage of the life cycle to progress to the next stage. Reversal of DAF-15 depletion permits arrested animals to recover to continue development. Our results are consistent with TORC1 functioning as a developmental checkpoint that governs the decision of the animal to progress through development.


A multicentre, prospective, randomised, double-blind study to measure the treatment effectiveness of abobotulinum A (AboBTXA) among women with refractory interstitial cystitis/bladder pain syndrome.
To determine if abobotulinumtoxin A (AboBTXA) is an effective treatment for interstitial cystitis/bladder pain syndrome (IC/BPS). We performed a double-blind study of 54 women with severe, refractory IC from three referral centres whom we randomly allocated to treatment with hydrodistension + injection of normal saline or to hydrodistension + injection with AboBTXA. The O'Leary-Sant questionnaire consists of problem (OLS-PI) and symptom (OLS-PI) index scores, and bladder diary data were compared between AboBTXA and control patients at baseline and at 3 months of follow-up. Measurements were made beyond 3 months, but no further randomised comparison was possible due to the ability of nonresponsive patients in either group to have AboBTXA treatment. Complete data were available in 50 patients, and in both groups, OLS questionnaires showed improvement at 3 months. Only the OLS-PI was improved in the AboBTXA group (p = 0.04). At 3 months, no difference was found in either OLS-SI or total OLS score. Twelve patients had urinary tract infection (UTI) treated during the follow-up period, which confounded results. In the 38 patients without UTI, there was improvement in total OLS score (p = 0.02), OLS-PI (0.08), and OLS-SI (p = 0.008) for the AboBTXA group at 3 months. Only five AboBTXA compared with two control patients had a 50% reduction in OLS score. For chronic refractory IC/BPS patients, AboBTXA was associated with no overall improvement in total OLS score, although significant benefit was noted in a small number of patients. The absence of posttreatment UTI was associated with a better response to AboBTXA.


A survey of leptospirological studies carried out on the Cape Verde Islands.
The authors subjected 611 human sera, 1295 sera of domestic and 211 sera of wild living animals from two islands of the Cape Verde archipelago to serological tests for leptospirosis. Among the human sera 7.2% reacted positively. Antibodies to L. bratislava of the serogroup Australis were most frequent (59.1% of all positive sera). Antibodies to L. icterohaemorrhagiae were found in 1.3% of the sera tested, to L. sejroe in 1.1%, to L. canicola in 0.7%, to L. pyrogenes in 0.3% and to L. pomona, L. bulgarica and L. grippotyphosa in 0.2% each. The existence of natural foci of leptospiroses could not be proved since tests of 211 wild-living mammals were all negative. The investigation of domestic animal sera showed a positivity of 3.1% with goats being positive most frequently (85% of all positive sera), followed by donkeys with 3.3% and cattle with 2.0%. Sera of dogs, sheep and pigs were all negative. The results indicate that anthropurgic foci exist on the Cape Verde Islands.


Performance of the ASSIGN cardiovascular disease risk score on a UK cohort of patients from general practice.
To evaluate the performance of ASSIGN against the Framingham equations for predicting 10 year risk of cardiovascular disease in a UK cohort of patients from general practice and to make the evaluation comparable to an independent evaluation of QRISK on the same cohort. Prospective open cohort study. Setting 288 practices from England and Wales contributing to The Health Improvement Network (THIN) database. Patients registered with 288 UK practices for some period between January 1995 and March 2006. The number of records available was 1,787,169. First diagnosis of myocardial infarction, coronary heart disease, stroke and transient ischaemic attacks recorded. Methods We implemented the Anderson Framingham Coronary Heart Disease and Stroke models, ASSIGN, and a more recent Framingham Cox proportional-hazards model and analysed their calibration and discrimination. Calibration showed that all models tested over-estimated risk particularly for men. ASSIGN showed better discrimination with higher AUROC (0.756/0.792 for men/women), D statistic (1.35/1.58 for men/women), and R²(30.47%/37.39% for men/women). The performance of ASSIGN was comparable to that of QRISK on the same cohort. Models agreed on 93-97% of categorical (high/lower) risk assessments and when they disagreed, ASSIGN was often closer to the estimated Kaplan-Meier incidence. ASSIGN also provided a steeper gradient of deprivation and discriminated between those with and without recorded family history of CVD. The estimated incidence was twice/three times as high for women/men with a recorded family history of CVD. For systematic CVD risk assessment all models could usefully be applied, but ASSIGN improved on the gradient of deprivation and accounted for recorded family history whereas the Framingham equations did not. However, all models display relatively low specificity and sensitivity. An additional conclusion is that the recording of family history of CVD in primary care databases needs to improve given its importance in risk assessment.


Coordination ability determined transition metal ions substitution of Tb in Tb-Asp fluorescent nanocrystals and a facile ions-detection approach.
Although the synthesis and fluorescent properties of lanthanide-amino acid complex nanostructures have been investigated extensively, limited studies have been reported on metal ions' substitution ability for the lanthanide ions in the complex and their effect on the fluorescent property. In this study, taking biocompatible Tb-aspartic acid (Tb-Asp) complex nanocrystals as a model, the substitution mechanism of metal ions, particularly transition metals, for Tb ions in Tb-Asp nanocrystals and the change in the fluorescent property of the Tb-Asp nanocrystals after substitution were systematically investigated. The experimental results illustrated that metal ions with higher electronegativity, higher valence, and smaller radius possess stronger ability for Tb ions' substitution in Tb-Asp nanocrystals. Based on the effect of substituting ions' concentration on the fluorescent property of Tb-Asp, a facile method for copper ions detection with high sensitivity was proposed by measuring the fluorescent intensity of Tb-Asp nanocrystals' suspensions containing different concentrations of copper ions. The good biocompatibility, great convenience of synthesis and sensitive detection ability make Tb-Asp nanocrystals a very low cost and effective material for metal ions detection, which also opens a new door for practical applications of metal-Asp coordinated nanocrystals.


Periodic mechanical stress activates MEK1/2-ERK1/2 mitogenic signals in rat chondrocytes through Src and PLCγ1.
The mitogenic effects of periodic mechanical stress on chondrocytes have been studied extensively but the mechanisms whereby chondrocytes sense and respond to periodic mechanical stress remain a matter of debate. We explored the signal transduction pathways of chondrocyte proliferation and matrix synthesis under periodic mechanical stress. In particular, we sought to identify the role of the MEK1/2-ERK1/2 signaling pathway in chondrocyte proliferation and matrix synthesis following cyclic physiologic mechanical compression. Under periodic mechanical stress, both rat chondrocyte proliferation and matrix synthesis were significantly increased (P < 0.05) and were associated with increases in the phosphorylation of Src, PLCγ1, MEK1/2, and ERK1/2 (P < 0.05). Pretreatment with the MEK1/2-ERK1/2 selective inhibitor, PD98059, and shRNA targeted to ERK1/2 reduced periodic mechanical stress-induced chondrocyte proliferation and matrix synthesis (P < 0.05), while the phosphorylation levels of Src-Tyr418 and PLCγ1-Tyr783 were not inhibited. Proliferation, matrix synthesis and phosphorylation of MEK1/2-Ser217/221 and ERK1/2-Thr202/Tyr204 were inhibited after pretreatment with the PLCγ1 inhibitor U73122 in chondrocytes in response to periodic mechanical stress (P < 0.05), while the phosphorylation site of Src-Tyr418 was not affected. Inhibition of Src activity with PP2 and shRNA targeted to Src abrogated chondrocyte proliferation and matrix synthesis (P < 0.05) and attenuated PLCγ1, MEK1/2 and ERK1/2 activation in chondrocytes subjected to periodic mechanical stress (P < 0.05). These findings suggest that periodic mechanical stress promotes chondrocyte proliferation and matrix synthesis in part through the Src-PLCγ1-MEK1/2-ERK1/2 signaling pathway, which links these three important signaling molecules into a mitogenic cascade.


The Arf6 activator Efa6/PSD3 confers regional specificity and modulates ethanol consumption in Drosophila and humans.
Ubiquitously expressed genes have been implicated in a variety of specific behaviors, including responses to ethanol. However, the mechanisms that confer this behavioral specificity have remained elusive. Previously, we showed that the ubiquitously expressed small GTPase Arf6 is required for normal ethanol-induced sedation in adult Drosophila. Here, we show that this behavioral response also requires Efa6, one of (at least) three Drosophila Arf6 guanine exchange factors. Ethanol-naive Arf6 and Efa6 mutants were sensitive to ethanol-induced sedation and lacked rapid tolerance upon re-exposure to ethanol, when compared with wild-type flies. In contrast to wild-type flies, both Arf6 and Efa6 mutants preferred alcohol-containing food without prior ethanol experience. An analysis of the human ortholog of Arf6 and orthologs of Efa6 (PSD1-4) revealed that the minor G allele of single nucleotide polymorphism (SNP) rs13265422 in PSD3, as well as a haplotype containing rs13265422, was associated with an increased frequency of drinking and binge drinking episodes in adolescents. The same haplotype was also associated with increased alcohol dependence in an independent European cohort. Unlike the ubiquitously expressed human Arf6 GTPase, PSD3 localization is restricted to the brain, particularly the prefrontal cortex (PFC). Functional magnetic resonance imaging revealed that the same PSD3 haplotype was also associated with a differential functional magnetic resonance imaging signal in the PFC during a Go/No-Go task, which engages PFC-mediated executive control. Our translational analysis, therefore, suggests that PSD3 confers regional specificity to ubiquitous Arf6 in the PFC to modulate human alcohol-drinking behaviors.


Masked hypertension in diabetes mellitus: treatment implications for clinical practice.
Although distinguishing features of masked hypertension in diabetics are well known, the significance of antihypertensive treatment on clinical practice decisions has not been fully explored. We analyzed 9691 subjects from the population-based 11-country International Database on Ambulatory Blood Pressure in Relation to Cardiovascular Outcomes. Prevalence of masked hypertension in untreated normotensive participants was higher (P<0.0001) among 229 diabetics (29.3%, n=67) than among 5486 nondiabetics (18.8%, n=1031). Over a median of 11.0 years of follow-up, the adjusted risk for a composite cardiovascular end point in untreated diabetic-masked hypertensives tended to be higher than in normotensives (hazard rate [HR], 1.96; 95% confidence interval [CI], 0.97-3.97; P=0.059), similar to untreated stage 1 hypertensives (HR, 1.07; CI, 0.58-1.98; P=0.82), but less than stage 2 hypertensives (HR, 0.53; CI, 0.29-0.99; P=0.048). In contrast, cardiovascular risk was not significantly different in antihypertensive-treated diabetic-masked hypertensives, as compared with the normotensive comparator group (HR, 1.13; CI, 0.54-2.35; P=0.75), stage 1 hypertensives (HR, 0.91; CI, 0.49-1.69; P=0.76), and stage 2 hypertensives (HR, 0.65; CI, 0.35-1.20; P=0.17). In the untreated diabetic-masked hypertensive population, mean conventional systolic/diastolic blood pressure was 129.2 ± 8.0/76.0 ± 7.3 mm Hg, and mean daytime systolic/diastolic blood pressure 141.5 ± 9.1/83.7 ± 6.5 mm Hg. In conclusion, masked hypertension occurred in 29% of untreated diabetics, had comparable cardiovascular risk as stage 1 hypertension, and would require considerable reduction in conventional blood pressure to reach daytime ambulatory treatment goal. Importantly, many hypertensive diabetics when receiving antihypertensive therapy can present with normalized conventional and elevated ambulatory blood pressure that mimics masked hypertension.


Esculetin (dihydroxycoumarin) inhibits the production of matrix metalloproteinases in cartilage explants, and oral administration of its prodrug, CPA-926, suppresses cartilage destruction in rabbit experimental osteoarthritis.
To investigate the in vitro effects of 6,7-dihydroxycoumarin (esculetin) on the production of matrix metalloproteinases (MMP) in rabbit articular cartilage, and the in vivo effects of orally administered CPA-926, a prodrug of esculetin, on cartilage destruction in rabbit experimental osteoarthritis (OA). In vitro studies were performed using rabbit articular cartilage explants. Esculetin 10-100 microM was added to cartilage explants in the presence or absence of interleukin 1alpha (IL-1alpha). Effects of esculetin on cartilage metabolism were assessed. Proteoglycan release into medium was determined by dye precipitation with 1,9-dimethylmethylene blue, synthesis of proMMP-1 (interstitial procollagenase) and proMMP-3 (prostromelysin 1) by Western blotting, and collagen degradation activity using FITC labeled collagen. In vivo experimental OA was induced in the knee joints of 15 Japanese adult white rabbits by partial lateral meniscectomy. Ten rabbits were orally administered 200 or 400 mg/kg/day of CPA-926 from the day of surgery for 14 days. The size of the macroscopic erosive area on the femoral condyle and tibial plateau was measured, and cartilage destruction was histologically evaluated. Collagenolytic activities in synovial fluid were measured using FITC labeled collagen as a substrate. In vitro, esculetin inhibited the IL-1alpha induced release of proteoglycan into the medium in a dose dependent manner. The collagenolytic activities in cartilage explant medium induced by IL-1alpha were also suppressed with the addition of 33-100 microM esculetin (p = 0.0209 at 33 and 100 microM, p = 0.0202 at 66 microM). Western blotting of cartilage explant medium showed a decrease in the levels of proMMP-1 and proMMP-3 in the medium by treatment with esculetin. In vivo: At 14 days after surgery, the femoral condyle and tibial plateau in the control group showed macroscopic erosions of cartilage. Compared with the control group, the rabbits treated with CPA-926 at the dose of 400 mg/kg exhibited reduction of the size of the erosive area on the tibial plateau (p = 0.009). Histological evaluation indicated protection against the development of destructive changes in the tibial plateau cartilage at a dose of 200 mg/kg (p = 0.0442) and 400 mg/kg (p = 0.0446) of CPA-926. These results indicate that esculetin inhibits matrix degradation in rabbit joint cartilage explants through the suppression of MMP synthesis, secretion, or activity. Prophylactic administration of its prodrug, CPA-926, appears to provide some protection against cartilage destruction in a short term rabbit experimental OA model.


Primary screening for cervical cancer based on high-risk human papillomavirus (HPV) detection and HPV 16 and HPV 18 genotyping, in comparison to cytology.
The objective of the present study is to assess the performance of a high-risk human papillomavirus (HR-HPV) DNA test with individual HPV-16/HPV-18 genotyping as a method for primary cervical cancer screening compared with liquid-based cytology (LBC) in a population of Greek women taking part in routine cervical cancer screening. The study, conducted by the "HEllenic Real life Multicentric cErvical Screening" (HERMES) study group, involved the recruitment of 4,009 women, aged 25-55, who took part in routine cervical screening at nine Gynecology Departments in Greece. At first visit cervical specimens were collected for LBC and HPV testing using the Roche Cobas 4800 system. Women found positive for either cytology or HPV were referred for colposcopy, whereas women negative for both tests will be retested after three years. The study is ongoing and the results of the first screening round are reported herein. Valid results for cytology and HPV testing were obtained for 3,993 women. The overall prevalence of HR-HPV was 12.7%, of HPV-16 2.7% and of HPV-18 1.4%. Of those referred for colposcopy, cervical intraepithelial neoplasia grade 2 or worse (CIN2+) was detected in 41 women (1.07%). At the threshold of CIN2+, cytology [atypical squamous cells of undetermined significance (ASC-US) or worse] and HPV testing showed a sensitivity of 53.7% and 100% respectively, without change between age groups. Cytology and HPV testing showed specificity of 96.8% and 90.3% respectively, which was increased in older women (≥30) in comparison to younger ones (25-29). Genotyping for HPV16/18 had similar accuracy to cytology for the detection of CIN2+ (sensitivity: 58.5%; specificity 97.5%) as well as for triage to colposcopy (sensitivity: 58.5% vs 53.7% for cytology). HPV testing has much better sensitivity than cytology to identify high-grade cervical lesions with slightly lower specificity. HPV testing with individual HPV-16/HPV-18 genotyping could represent a more accurate methodology for primary cervical cancer screening in comparison to liquid-based cytology, especially in older women.


Routine examination of the newborn: the EMREN study. Evaluation of an extension of the midwife role including a randomised controlled trial of appropriately trained midwives and paediatric senior house officers.
To assess the implications and cost-effectiveness of extending the role of midwives to include the routine (24-hour) examination of the healthy newborn usually carried out by junior doctors. The study included a prospective randomised controlled trial (RCT) with mother and baby dyads randomised to either senior house officer (SHO) or midwife for the routine examination of the newborn. Midwives and SHOs were also videoed while performing the examinations and the videos were rated by an independent consultant and senior midwife. In addition extensive interviews, surveys, consultations and assessments were carried out. A District General Hospital (for the RCT), a London Teaching Hospital, general practices and mothers' homes (for interviews); questionnaires were sent to all maternity units in England (for the National Survey). A total of 826 mother and baby dyads in a District General Hospital in south-east England. Midwives and SHOs, as well as midwifery managers, paediatric consultants; general practitioners (GPs) and representatives of key organisations. A routine examination of a newborn baby was carried out at about 24 hours from birth and a further examination for half the babies in each group, at 10-days at home by the community midwife. Referrals assessed as appropriate and as major or minor by three independent consultants. Problems identified during the first year of life assessed as identifiable at 24 hours. Quality assessment by video against an agreed written proforma. Maternal satisfaction. Opinion of professionals and mothers about aspects of the examination. There was no statistical difference between SHO and midwife examinations in appropriate referral rates to hospital or community or in inappropriate referral rates to hospital. Videoed assessments were assessed as carried out more appropriately by the midwives than by the SHOs. Overall maternal satisfaction was high and higher when a midwife rather than an SHO examined. Few new health problems were identified at the 10-day examination. From the National Survey, it was estimated that about 2% of babies in England are examined by a midwife. If midwives were to examine all babies where there were no complications of birth or antenatal history, there would be savings of about 2 pounds per baby born, equivalent to savings of 1.2 pounds million nationally. Were midwives to examine all babies on normal wards savings would increase to about 4.30 pounds per baby born or 2.5 million pounds nationally. Representatives of the professional bodies were of the opinion that having trained midwives carrying out the examination would be valuable. All component aspects of the study were consistent in showing benefits or at least no significant barriers to suitably qualified, trained midwives carrying out the examinations. Developing the role of the midwife to include examination of the newborn is likely to result in improved quality of examinations and higher satisfaction from mothers. It would slightly reduce overall health service costs, with some increased resources needed by midwifery departments, and some decrease in resource needs of paediatric departments. There is a need for further research into the value of the examination being carried out at home rather than in hospital; the overall unsatisfactory quality of the examination of the hips; and appropriate inclusion criteria for which babies' midwives should examine.


Pref-1 preferentially inhibits heat production in brown adipose tissue.
In mammals there are two types of adipocytes with opposing functions. Brown adipocytes are characterized by a high number of mitochondria and are specialized for heat production (thermogenesis), expressing thermogenic genes such as UCP1 (uncoupling protein 1). White adipocytes, on the other hand, store energy. Although many key regulators in the differentiation of white adipocytes have been established, our current knowledge on the same proteins in brown adipogenesis is lagging behind. One example is Pref-1 (pre-adipocyte factor-1), which maintains white pre-adipocytes in an undifferentiated state, but is only poorly characterized in the brown pre-adipocyte lineage. In this issue of the Biochemical Journal, Armengol et al. now shed new light on the role and regulation of Pref-1 in brown pre-adipocytes. First, Pref-1 specifically inhibits the thermogenic gene programme in brown pre-adipocytes. Secondly, they identified the transcription factor C/EBPδ (CCAAT/enhancer-binding protein δ) as a direct positive regulator of Pref-1 expression, whereas this protein does not fulfil this role in white adipogenesis. Taken together, these findings indicate that specific manipulation of brown adipocyte differentiation and/or function without interfering with their white adipocyte counterparts may be possible, which may open up new therapeutic ways to combat obesity-associated health problems.


ASH1 mRNA anchoring requires reorganization of the Myo4p-She3p-She2p transport complex.
One mechanism by which cells post-transcriptionally regulate gene expression is via intercellular and intracellular sorting of mRNA. In Saccharomyces cerevisiae, the localization of ASH1 mRNA to the distal tip of budding cells results in the asymmetric sorting of Ash1p to daughter cell nuclei. Efficient localization of ASH1 mRNA depends upon the activity of four cis-acting localization elements and also upon the activity of trans-factors She2p, She3p, and Myo4p. She2p, She3p, and Myo4p have been proposed to form an ASH1 mRNA localization particle. She2p directly and specifically binds each of the four ASH1 cis-acting localization elements, whereas She3p has been hypothesized to function as an adaptor by recruiting the She2p-mRNA complex to Myo4p, a type V myosin. The Myo4p-She3p-She2p heterotrimeric protein complex has been proposed to localize mRNA to daughter cells using polarized actin cables. Here we demonstrate that whereas the predicted Myo4p-She3p-She2p heterotrimeric complex forms in vivo, it represents a relatively minor species compared with the Myo4p-She3p complex. Furthermore, contrary to a prediction of the heterotrimeric complex model for ASH1 mRNA localization, ASH1 mRNA artificially tethered to She2p is not localized. Upon closer examination, we found that mRNA tightly associated with She2p is transported to daughter cells but is not properly anchored at the bud tip. These results are consistent with a model whereby anchoring of ASH1 mRNA requires molecular remodeling of the Myo4p-She3p-She2p heterotrimeric complex, a process that is apparently altered when mRNA is artificially tethered to She2p.


Method of delivery and pregnancy outcomes in Asia: the WHO global survey on maternal and perinatal health 2007-08.
There has been concern about rising rates of caesarean section worldwide. This Article reports the third phase of the WHO global survey, which aimed to estimate the rate of different methods of delivery and to examine the relation between method of delivery and maternal and perinatal outcomes in selected facilities in Africa and Latin America in 2004-05, and in Asia in 2007-08. Nine countries participated in the Asia global survey: Cambodia, China, India, Japan, Nepal, Philippines, Sri Lanka, Thailand, and Vietnam. In each country, the capital city and two other regions or provinces were randomly selected. We studied all women admitted for delivery during 3 months in institutions with 6000 or fewer expected deliveries per year and during 2 months in those with more than 6000 deliveries. We gathered data for institutions to obtain a detailed description of the health facility and its resources for obstetric care. We obtained data from women's medical records to summarise obstetric and perinatal events. We obtained data for 109 101 of 112 152 deliveries reported in 122 recruited facilities (97% coverage), and analysed 107 950 deliveries. The overall rate of caesarean section was 27.3% (n=29 428) and of operative vaginal delivery was 3.2% (n=3465). Risk of maternal mortality and morbidity index (at least one of: maternal mortality, admission to intensive care unit [ICU], blood transfusion, hysterectomy, or internal iliac artery ligation) was increased for operative vaginal delivery (adjusted odds ratio 2.1, 95% CI 1.7-2.6) and all types of caesarean section (antepartum without indication 2.7, 1.4-5.5; antepartum with indication 10.6, 9.3-12.0; intrapartum without indication 14.2, 9.8-20.7; intrapartum with indication 14.5, 13.2-16.0). For breech presentation, caesarean section, either antepartum (0.2, 0.1-0.3) or intrapartum (0.3, 0.2-0.4), was associated with improved perinatal outcomes, but also with increased risk of stay in neonatal ICU (2.0, 1.1-3.6; and 2.1, 1.2-3.7, respectively). To improve maternal and perinatal outcomes, caesarean section should be done only when there is a medical indication. US Agency for International Development (USAID); UNDP/UNFPA/WHO/World Bank Special Programme of Research, Development and Research Training in Human Reproduction (HRP), WHO, Switzerland; Ministry of Health, Labour and Welfare of Japan; Ministry of Public Health, China; and Indian Council of Medical Research.


Phylodynamics and dispersal of HRSV entails its permanence in the general population in between yearly outbreaks in children.
Human respiratory syncytial virus (HRSV) is one of the major etiologic agents of respiratory tract infections among children worldwide. Here through a comprehensive analysis of the two major HRSV groups A and B (n=1983) which comprise of several genotypes, we present a complex pattern of population dynamics of HRSV over a time period of 50 years (1956-2006). Circulation pattern of HRSV revealed a series of expansions and fluctuations of co-circulating lineages with a predominance of HRSVA. Positively selected amino acid substitutions of the G glycoprotein occurred upon population growth of GB3 with a 60-nucleotide insertion (GB3 Insert), while other genotypes acquired substitutions upon both population growth and decrease, thus possibly reflecting a role for immune selected epitopes in linkage to the traced substitution sites that may have important relevance for vaccine design. Analysis evidenced the co-circulation and predominance of distinct HRSV genotypes in Brazil and suggested a year-round presence of the virus. In Brazil, GA2 and GA5 were the main culprits of HRSV outbreaks until recently, when the GB3 Insert became highly prevalent. Using Bayesian methods, we determined the dispersal patterns of genotypes through several inferred migratory routes. Genotypes spread across continents and between neighboring areas. Crucially, genotypes also remained at any given region for extended periods, independent of seasonal outbreaks possibly maintained by re-infecting the general population.


Prototype development and field-test results of an adaptive multiresolution PANOPTES imaging architecture.
The design, development, and field-test results of a visible-band, folded, multiresolution, adaptive computational imaging system based on the Processing Arrays of Nyquist-limited Observations to Produce a Thin Electro-optic Sensor (PANOPTES) concept is presented. The architectural layout that enables this imager to be adaptive is described, and the control system that ensures reliable field-of-view steering for precision and accuracy in subpixel target registration is explained. A digital superresolution algorithm introduced to obtain high-resolution imagery from field tests conducted in both nighttime and daytime imaging conditions is discussed. The digital superresolution capability of this adaptive PANOPTES architecture is demonstrated via results in which resolution enhancement by a factor of 4 over the detector Nyquist limit is achieved.


Gender Parity in Critical Care Medicine.
Clinical practice guidelines are systematically developed statements to assist practitioner and patient decisions about appropriate health care for specific clinical circumstances. These documents inform and shape patient care around the world. In this Perspective we discuss the importance of diversity on guideline panels, the disproportionately low representation of women on critical care guideline panels, and existing initiatives to increase the representation of women in corporations, universities, and government. We propose five strategies to ensure gender parity within critical care medicine.


Severe xanthomatosis associated with familial apolipoprotein E deficiency.
To present the clinical, dermatological, and histological features of a patient with generalised xanthomatosis, familial apolipoprotein (apo) E deficiency, and unusual type III hyperlipoproteinaemia (HLP). The underlying molecular defect was disclosed using molecular biological techniques. The unusual xanthomas were histologically analysed and the morphology of the abnormal lipoprotein particles examined using electron microscopy. A 10 base pair deletion in exon 4 of the proband's apo epsilon gene (base pairs 4037-4046 coding for amino acids 209-212 of the mature protein) was identified. This is predictive for a reading frameshift encoding a premature stop (TGA) in codon 229. The mutation is responsible for delayed catabolism of atherogenic lipoprotein remnants, lipid storage in monocyte/macrophages, and phenotypic expression of xanthomatosis early in life. Familial apo E deficiency is a rare genetic disease which offers the unique opportunity to study the impact of apo E on lipoprotein metabolism and development of atherosclerosis in humans.


Effects of extreme hemodilution during cardiac surgery on cognitive function in the elderly.
Strategies for neuroprotection including hypothermia and hemodilution have been routinely practiced since the inception of cardiopulmonary bypass. Yet postoperative neurocognitive deficits that diminish the quality of life of cardiac surgery patients are frequent. Because there is uncertainty regarding the impact of hemodilution on perioperative organ function, the authors hypothesized that extreme hemodilution during cardiac surgery would increase the frequency and severity of postoperative neurocognitive deficits. Patients undergoing coronary artery bypass grafting surgery were randomly assigned to either moderate hemodilution (hematocrit on cardiopulmonary bypass >or=27%) or profound hemodilution (hematocrit on cardiopulmonary bypass of 15-18%). Cognitive function was measured preoperatively and 6 weeks postoperatively. The effect of hemodilution on postoperative cognition was tested using multivariable modeling accounting for age, years of education, and baseline levels of cognition. After randomization of 108 patients, the trial was terminated by the Data Safety and Monitoring Board due to the significant occurrence of adverse events, which primarily involved pulmonary complications in the moderate hemodilution group. Multivariable analysis revealed an interaction between hemodilution and age wherein older patients in the profound hemodilution group experienced greater neurocognitive decline (P = 0.03). In this prospective, randomized study of hemodilution during cardiac surgery with cardiopulmonary bypass in adults, the authors report an early termination of the study because of an increase in adverse events. They also observed greater neurocognitive impairment among older patients receiving extreme hemodilution.


Increased Mycobacterium tuberculosis antigen-induced gene expression of interferon-gamma, tumor necrosis factor alpha and interleukin-6 in patients with diabetes.
Pakistan ranks fifth in high tuberculosis (TB)-burden countries and seventh among countries with high prevalence rates of diabetes mellitus (DM). DM is a risk factor for TB and worsens disease outcomes. Furthermore, Mycobacterium tuberculosis (MTB) infection can induce glucose intolerance and worsen glycemic control in diabetes. Suppressor of cytokine signaling (SOCS)-1 and -3 molecules regulate cytokine signaling and are important in maintaining an immune balance. In TB, interleukin (IL)-6 upregulation induces SOCS3, which is also a negative regulator of insulin signaling. This research focuses on the mechanism by which SOCS1 and SOCS3 affect insulin resistance and increased susceptibility to TB. We studied gene expression in peripheral blood cells of patients with diabetes (n=10) and healthy endemic controls (EC, n=11) both with and without MTB infection. Mycobacterial antigen (PPD) and mitogen-stimulated SOCS1, SOCS3, interferon-gamma (IFN-γ), IL-6, and tumor necrosis factor alpha (TNFα) mRNA expression levels were determined using real-time polymerase chain reaction. MTB antigen-stimulated mRNA levels of IFN-γ was 10-fold higher, SOCS1 was 4 times greater, TNFα was 10-fold higher, and IL-6 was 2-fold greater in patients with DM than in ECs. Overall levels of PPD-stimulated IL-6 was higher in patients with DM than in ECs (p=.036). Mitogen-induced mRNA levels of IFN-γ were 30-fold higher, SOCS3 was 20 fold higher, and SOCS1 was 4-fold higher in patients with DM than in ECs. Increased proinflammatory cytokine production in response to MTB antigens in diabetes would lead to exacerbated pathology and reduced inflammatory control at the site of MTB infection. This would in turn hamper the resolution of inflammation, resulting in unfavorable disease outcomes.


Risks of self-medication practices.
Self-medication is defined as the selection and use of medicines by individuals (or a member of the individuals' family) to treat self-recognized or self-diagnosed conditions or symptoms. Several benefits have been linked to appropriate self-medication, among them: increased access to medication and relief for the patient, the active role of the patient in his or her own health care, better use of physicians and pharmacists skills and reduced (or at least optimized) burden of governments due to health expenditure linked to the treatment of minor health conditions However, self-medication is far from being a completely safe practice, in particular in the case of non-responsible self-medication. Potential risks of self-medication practices include: incorrect self-diagnosis, delays in seeking medical advice when needed, infrequent but severe adverse reactions, dangerous drug interactions, incorrect manner of administration, incorrect dosage, incorrect choice of therapy, masking of a severe disease and risk of dependence and abuse. In this short review the author analyzes recent literature on some of the most important dangers related to self-medication practices, particularly: polypharmacy and drug interactions, medications abuse or dependence, misdiagnosis and incorrect choice of treatment. The author also proposes measures that could be adopted in order to solve or improve these issues.


Fine-needle aspiration cytology accuracy with palpable gynecologic neoplasms.
From 1984 to 1988, 62 fine-needle aspirations (FNA) were performed on palpable lesions in 59 gynecologic oncology patients at the UCLA Medical Center. Sites of aspiration included abdomen, cervix, vagina, superficial lymph nodes, and pelvic masses. Confirmatory open biopsy (41) or adequate clinical follow-up (17) was obtained in 55 patients (58 aspirates). FNA correctly established the diagnosis of malignancy in 19 of 26 (73%) biopsied patients. The predictive value for a positive test was 100%, and the predictive value for a negative test was 82%. Initial surgical biopsy had been incorrectly benign in 4 of these patients who were shown subsequently to have malignant tumors by FNA and clinical findings. In 7 patients, FNA failed to diagnose the malignancy found by open biopsy. Two of the false-negative FNAs were insufficient and five were in masses where palpation was inadequate. In 17 patients who were followed clinically without open biopsy, FNA correctly predicted the subsequent clinical course in 15 (88%). There were no false-positive FNA diagnoses obtained when cytologic results were correlated with both clinical outcome and surgical biopsy. Aspiration cytology has a specificity of 100% and a sensitivity of 65%. A negative FNA obtained from a clinically suspicious lesion should be followed by a repeat aspiration or surgical biopsy.


Real-time dual-loop electric current measurement for label-free nanofluidic preconcentration chip.
An electrokinetic trapping (EKT)-based nanofluidic preconcentration device with the capability of label-free monitoring trapped biomolecules through real-time dual-loop electric current measurement was demonstrated. Universal current-voltage (I-V) curves of EKT-based preconcentration devices, consisting of two microchannels connected by ion-selective channels, are presented for functional validation and optimal operation; universal onset current curves indicating the appearance of the EKT mechanism serve as a confirmation of the concentrating action. The EKT mechanism and the dissimilarity in the current curves related to the volume flow rate (Q), diffusion coefficient (D), and diffusion layer (DL) thickness were explained by a control volume model with a five-stage preconcentration process. Different behaviors of the trapped molecular plug were categorized based on four modes associated with different degrees of electroosmotic instability (EOI). A label-free approach to preconcentrating (bio)molecules and monitoring the multibehavior molecular plug was demonstrated through real-time electric current monitoring, rather than through the use of microscope images.


Epidermal growth factor receptor T790M testing in progressed lung cancer: A review of sensitive methods for analysis of tissue and liquid biopsy samples.
Lung cancer is one of the major causes of mortality worldwide and is on the rise in India. The identification of epidermal growth factor receptor (EGFR) mutations in nonsmall cell lung cancer (NSCLC) has paved the way for personalized therapy in lung cancer with EGFR-tyrosine kinase inhibitors (TKIs). Despite the proven efficacy of EGFR-TKIs in patients harboring EGFR mutations, their clinical utility is limited by the development of acquired resistance mechanisms by the tumor cells. T790M mutation accounts for 60% of all resistance mechanisms to EGFR TKIs and is responsible for treatment failure with first- and second-generation TKIs. With the development of novel therapeutic agents such as osimertinib to overcome this resistance mechanism, it is essential to detect patients harboring T790M mutation. There are several limitations with the use of tissue biopsy specimens for molecular testing such as poor quality and quantity of sample, tumor heterogeneity, occurrence of complications, and issues with repeat biopsy. Liquid biopsy offers a noninvasive approach that can be used for diagnostic purposes as well as for monitoring treatment response and evaluation of resistance mechanisms. This review focuses on the methods for molecular testing of tissue and liquid biopsy specimens for EGFR mutations, particularly EGFR T790M mutation.


Initial genome scan of the NIMH genetics initiative bipolar pedigrees: chromosomes 4, 7, 9, 18, 19, 20, and 21q.
An initial genome scan was performed on 540 individuals from 97 families segregating bipolar disorder, collected through the National Institutes of Mental Health Genetics Initiative. We report here affected-sib-pair (ASP) data on 126 marker loci (approximately 68,000 genotypes) mapping to chromosomes 4, 7, 9, 18, 19, 20, and 21q, under three affection status models. Modest increases in identical-by-descent (IBD) allele sharing were found at the following loci: D4S2397 and D4S391 (P < 0.05) on 4p, D4S1647 (P < 0.05) on 4q, D7S1802 and D7S1869 (low P = 0.01) on 7p, D9S302 (P = 0.004) on 9q, and D20S604 on 20p and D20S173 on 20q (P < 0.05). In addition, five markers on 7q displayed increased IBD sharing (P = 0.046-0.002). Additional ASP analyses on chromosomes 18 and 21q marker data were performed using disease phenotype models defined previously. On chromosome 18, only D18S40 on 18p and D18S70 on 18q yielded a slight elevation in allele sharing (P = 0.02), implying that the reported linkages in these regions were not confirmed. On chromosome 21q, a cluster of markers within an approximately 9 cM interval: D21S1254, D21S65, D21S1440, and D21S1255 exhibited excess allele sharing (P = 0.041-0.008). Multilocus data on overlapping marker quartets, from D21S1265 to D21S1255, which were consistent with increased IBD sharing (P < 0.01, with a low of 0.0009), overlapped a broad interval of excess allele sharing reported previously, increasing support for a susceptibility locus for bipolar disorder on 21q.


How to engage small retail businesses in workplace violence prevention: Perspectives from small businesses and influential organizations.
Small retail businesses experience high robbery and violent crime rates leading to injury and death. Workplace violence prevention programs (WVPP) based on Crime Prevention Through Environmental Design reduce this risk, but low small business participation limits their effectiveness. Recent dissemination models of occupational safety and health information recommend collaborating with an intermediary organization to engage small businesses. Qualitative interviews with 70 small business operators and 32 representatives of organizations with small business influence were conducted to identify factors and recommendations for improving dissemination of a WVPP. Both study groups recommended promoting WVPPs through personal contacts but differed on other promotion methods and the type of influential groups to target. Small business operators indicated few connections to formal business networks. Dissemination of WVPPs to small businesses may require models inclusive of influential individuals (e.g., respected business owners) as intermediaries to reach small businesses with few formal connections.


Prenatal masculinization of the auditory system in infants: The MIREC-ID study.
Sex differences in inner-ear function are detectable in infants, notably through the measurement of otoacoustic emissions (OAEs). Prevailing theories posit that prenatal exposure to high levels of androgens in boys may weaken OAEs, and that this phenomenon may predominantly affect the right ear/left hemisphere (Geschwind-Galaburda (GG) hypothesis). Yet, actual tests of these models have been difficult to implement in humans. Here we examined the relationship between markers of fetal androgen exposure collected at birth (anogenital distances (AGD); penile length/width, areolar/scrotal/vulvar pigmentation) and at 6 months of age (2nd to 4th digit ratio (2D:4D)) with two types of OAEs, click-evoked OAEs (CEOAEs) and distortion-product OAEs (DPOAEs) (n = 49; 25 boys; 24 girls). We found that, in boys, scrotal pigmentation was inversely associated with the amplitude and reproducibility of CEOAEs in the right ear at 4 kHz, with trends also present in the same ear for mean CEOAE amplitude and CEOAE amplitude at 2 kHz. Penile length was inversely associated with the mean amplitude of DPOAEs in both the right and left ears, as well as with DPOAE amplitude in the right ear at 2 kHz and the reproducibility of CEOAEs in the left ear at 2.8 kHz. Finally, AGD-scrotum in boys was positively associated in boys with the amplitude of DPOAEs in the left ear at 2.8 kHz. Unexpectedly, there were no sex differences in the amplitude or reproducibility of OAEs, nor, in girls, any associations between androgenic markers and auditory function. Nonetheless, these findings, reported for the first time in a sample of human infants, support both the prenatal-androgen-exposure and GG models as explanations for the masculinization of auditory function in male infants.


A novel model of lethal Hendra virus infection in African green monkeys and the effectiveness of ribavirin treatment.
The henipaviruses, Hendra virus (HeV) and Nipah virus (NiV), are emerging zoonotic paramyxoviruses that can cause severe and often lethal neurologic and/or respiratory disease in a wide variety of mammalian hosts, including humans. There are presently no licensed vaccines or treatment options approved for human or veterinarian use. Guinea pigs, hamsters, cats, and ferrets, have been evaluated as animal models of human HeV infection, but studies in nonhuman primates (NHP) have not been reported, and the development and approval of any vaccine or antiviral for human use will likely require efficacy studies in an NHP model. Here, we examined the pathogenesis of HeV in the African green monkey (AGM) following intratracheal inoculation. Exposure of AGMs to HeV produced a uniformly lethal infection, and the observed clinical signs and pathology were highly consistent with HeV-mediated disease seen in humans. Ribavirin has been used to treat patients infected with either HeV or NiV; however, its utility in improving outcome remains, at best, uncertain. We examined the antiviral effect of ribavirin in a cohort of nine AGMs before or after exposure to HeV. Ribavirin treatment delayed disease onset by 1 to 2 days, with no significant benefit for disease progression and outcome. Together our findings introduce a new disease model of acute HeV infection suitable for testing antiviral strategies and also demonstrate that, while ribavirin may have some antiviral activity against the henipaviruses, its use as an effective standalone therapy for HeV infection is questionable.


Novel dual regulators of Pseudomonas aeruginosa essential for productive biofilms and virulence.
Gene regulation network in Pseudomonas aeruginosa is complex. With a relatively large genome (6.2 Mb), there is a significant portion of genes that are proven or predicted to be transcriptional regulators. Many of these regulators have been shown to play important roles in biofilm formation and maintenance. In this study, we present a novel transcriptional regulator, PA1226, which modulates biofilm formation and virulence in P. aeruginosa. Mutation in the gene encoding this regulator abolished the ability of P. aeruginosa to produce biofilms in vitro, without any effect on the planktonic growth. This regulator is also essential for the in vivo fitness and pathogenesis in both Drosophila melanogaster and BALB/c mouse lung infection models. Transcriptome analysis revealed that PA1226 regulates many essential virulence genes/pathways, including those involved in alginate, pili, and LPS biosynthesis. Genes/operons directly regulated by PA1226 and potential binding sequences were identified via ChIP-seq. Attempts to confirm the binding sequences by electrophoretic mobility shift assay led to the discovery of a co-regulator, PA1413, via co-immunoprecipitation assay. PA1226 and PA1413 were shown to bind collaboratively to the promoter regions of their regulons. A model is proposed, summarizing our finding on this novel dual-regulation system.


Discourse analysis: examining the potential for research in palliative care.
Qualitative research methods are an appropriate way to address questions on understandings, feelings and experiences of people in the context of palliative care. They make an assumption that language reflects objective reality. An alternative position is taken by discourse analysis methods, which are concerned with investigating how language and written texts are used to shape and influence behaviours, and provide ways of constructing the world. This paper argues that discourse analysis has benefits in revealing taken-for-granted and hidden aspects of palliative care. These advantages include revealing how common words and terms are used to invoke social practices, knowledge and power, which shape the meaning and delivery of palliative care in different contexts. Advice on conducting discourse analysis is provided, along with two exemplar studies. Finally, we indicate the potential contributions, the disadvantages and the ethical considerations required in using this approach to research.


More standing and just as productive: Effects of a sit-stand desk intervention on call center workers' sitting, standing, and productivity at work in the Opt to Stand pilot study.
This study evaluated the effects of sit-stand desks on workers' objectively and subjectively assessed sitting, physical activity, and productivity. This quasi-experimental study involved one intervention group (n = 16) and one comparison group (n = 15). Participants were call center employees from two job-matched teams at a large telecommunications company in Sydney, Australia (45% female, 33 ± 11 years old). Intervention participants received a sit-stand desk, brief training, and daily e-mail reminders to stand up more frequently for the first 2 weeks post-installation. Control participants carried out their usual work duties at seated desks. Primary outcomes were workday sitting and physical activity assessed using ActivPAL or ActiGraph devices and self-report questionnaires. Productivity outcomes were company-specific objective metrics (e.g., hold time, talking time, absenteeism) and subjective measures. Measurements were taken at baseline, 1, 4, and 19 weeks post-installation. Intervention participants increased standing time after 1 week (+ 73 min/workday (95% CI: 22, 123)) and 4 weeks (+ 96 min/workday (95% CI: 41, 150)) post-intervention, while control group showed no changes. Between-group differences in standing time at one and 4 weeks were + 78 (95% CI: 9, 147) and + 95 min/workday (95% CI: 15, 174), respectively. Sitting time in the intervention group changed by - 64 (95% CI: - 125, - 2), - 76 (95% CI: - 142, - 11), and - 100 min/workday (95% CI: - 172, - 29) at 1, 4, and 19 weeks post-installation, respectively, while the control group showed no changes. No changes were observed in productivity outcomes from baseline to follow-up in either group. Sit-stand desks can increase standing time at work in call center workers without reducing productivity.


Atomic Force Microscopy Identifying Fuel Pyrolysis Products and Directing the Synthesis of Analytical Standards.
Here we present a new method that integrates atomic force microscopy (AFM) with analytical tools such as high-performance liquid chromatography (HPLC) with diode-array ultraviolet-visible (UV) absorbance, and mass spectrometry (MS) along with synthetic chemistry. This allows the detection, identification, and quantification of novel polycyclic aromatic hydrocarbons (PAH) in complex molecular mixtures. This multidisciplinary methodology is employed to characterize the supercritical pyrolysis products of n-decane, a model fuel. The pyrolysis experiments result in a complex mixture of both unsubstituted as well as highly methylated PAH. We demonstrate the AFM-driven discovery of a novel compound, benz[ l]indeno[1,2,3- cd]pyrene, with the chemical structure assignment serving as input for the chemical synthesis of such molecule. The synthesis is verified by AFM, and the synthesized compound is used as a reference standard in analytical measurements, establishing the first-ever unequivocal identification and quantification of this PAH as a fuel product. Moreover, the high-resolution AFM analysis detected several five- to eight-ring PAH, which represents novel fuel pyrolysis and/or combustion products. This work provides a route to develop new analytical standards by symbiotically using AFM, chemical synthesis, and modern analytical tools.


Rice production with minimal irrigation and no nitrogen fertilizer by intensive use of treated municipal wastewater.
We designed a new cultivation system of rice with circulated irrigation to remove nitrogen from treated municipal wastewater effectively and assessed the possibility of nitrogen removal in the new system without any adverse effects on rice production through bench-scale experiments through two seasons. Overgrowth of the rice plant, which can lead to lodging and tasteless rice, was found in the first season probably because nitrogen supply based on standard practice in normal paddy fields was too much in the closed irrigation system. In the second season, therefore, the amount of treated wastewater initially applied to the system was reduced but this resulted in a considerably decreased yield. On the other hand, the taste of the rice was significantly improved. The two-season experiments revealed that the new system enabled rice production with minimal irrigation (approximately 50% on the yield base compared to normal paddy fields) and no nitrogen fertilizer. The system also achieved >95% removal of nitrogen from the treated wastewater used for circulated irrigation. The accumulation of harmful metals in the rice was not observed after one season of cultivation in the new system. The accumulation after cultivation using the same soil repeatedly for a longer time should be examined by further studies.


Update on Tetracycline Susceptibility of Pediococcus acidilactici Based on Strains Isolated from Swiss Cheese and Whey.
Bacterial strains used as starter cultures in the production of fermented foods may act as reservoirs for antibiotic resistance (AbR) genes. To avoid the introduction of such genes into the food chain, the presence of acquired AbR in bacterial strains added to food must be tested. Standard protocols and microbiological cut-off values have been defined to provide practitioners with a basis for evaluating whether their bacterial isolates harbor an acquired resistance to a given antibiotic. Here, we tested the AbR of 24 strains of Pediococcus acidilactici by using the standard protocol and microbiological cut-off values recommended by the European Food Safety Authority. Phenotypic data were complemented by searching for known AbR genes using an in silico analysis of whole genomes. The majority (54.2%) of the strains were able to grow at a tetracycline concentration above the defined cut-off, even though only one strain carried a known tetracycline resistance gene, tetM. The same strain also carried the AbR gene of an erythromycin resistance methylase, ermA, and displayed resistance toward clindamycin and erythromycin. Our results bolster the scarce data on the sensitivity of P. acidilactici to tetracycline and suggest that the microbiological cut-off recommended by the European Food Safety Authority for this antibiotic should be amended.


Parasternal infiltration composite breast augmentation.
The simultaneous combination of both fat grafting to the breast and mammary implants has been recently proposed as a useful technique in breast augmentation surgery. The purpose of this study was to evaluate the aesthetic benefits of selective parasternal fat grafting at the time of primary implant breast augmentation. Fifty-nine consecutive primary breast augmentation patients were studied retrospectively. Patients were divided into two groups: group 1 patients (n = 38) were treated only with breast implants, whereas group 2 patients (n = 21) received breast implants and parasternal fat grafting of 60 to 140 cc of adipose tissue. The length between the medial border of each breast, defined as the parasternal vertical aesthetic line, was measured preoperatively and postoperatively for both groups and compared through statistical analysis. The mean length between the parasternal vertical aesthetic lines in group 1 postoperatively was significantly higher, 2.26 ± 1.24 cm (p < 0.0001); whereas this length for group 2 was significantly lower after surgery, 0.60 ± 0.32 cm (p < 0.0001). No cysts or fat necroses were observed in group 2, presumably because of the low volume of fat used. Parasternal fat grafting performed simultaneously with breast augmentation is a safe procedure, and seems to provide a valuable cosmetic advantage by improving the medial transition zone of the breast implant with the presternal area. It prevents a "separated-breasts" deformity, which may produce unnatural results in implant-based breast augmentations, especially in thin patients.


Nonlinear optical properties of induced transmission filters.
The nonlinear optical (NLO) properties of induced transmission filters (ITFs) based on Ag are experimentally determined using white light continuum pump-probe measurements. The experimental results are supported using simulations based on the matrix transfer method. The magnitude of the NLO response is shown to be 30 times that of an isolated Ag film of comparable thickness. The impacts of design variations on the linear and NLO response are simulated. It is shown that the design can be modified to enhance the NLO response of an ITF by a factor of 2 or more over a perfectly matched ITF structure.


The human moment at work.
In the last decade or so, technological changes--mainly voice mail and e-mail--have made a lot of face-to-face interaction unnecessary. Face-to-face contact has also fallen victim to "virtuality"--many people work at home or are otherwise off-site. Indeed, most people today can't imagine life without such technology and the freedom it grants. But Edward Hallowell, a noted psychiatrist who has been treating patients with anxiety disorders--many of them business executives--for more than 20 years, warns that we are in danger of losing what he calls the human moment: an authentic psychological encounter that can happen only when two people share the same physical space. And, he believes, we may be about to discover the destructive power of its absence. The author relates stories of business-people who have dealt firsthand with the misunderstandings caused by an overreliance on technology. An e-mail message is misconstrued. Someone forwards a voice-mail message to the wrong people. A person takes offense because he was not included on a certain circulation list. Was it an accident? Often the consequences of such misunderstandings, taken individually, are minor. Over time, however, they take a larger toll--both on individuals and on the organizations they work for. The problem, however, is not insoluble. The author cites examples of people who have worked successfully to restore face-to-face contact in their organizations. The bottom line is that the strategic use of the human moment adds color to our lives and helps us build confidence and trust at work. We ignore it at our peril.


"This is streets ahead of what we used to do": staff perceptions of virtual clinical pharmacy services in rural and remote Australian hospitals.
The use of medications is the most common intervention in healthcare. However, unsafe medication practices and medication errors are a leading cause of injury and avoidable harm in healthcare systems across the world. A Virtual Clinical Pharmacy Service (VCPS) was introduced in rural and remote New South Wales public hospitals to support safe and effective use of medications. In this model clinical pharmacy services are delivered via a telehealth cart at the patient's bedside and through electronic medical and pharmaceutical record systems. The aim of this research was to understand healthcare staff perspectives of the VCPS and identify areas for improvement. A qualitative approach informed by Appreciative Inquiry was used to investigate healthcare staff perceptions of the VCPS. Focus group discussions (n = 15) with hospital staff and medical officers were conducted via videoconference at each study site. Focus groups explored issues, benefits and barriers 3 months after service implementation. Transcribed data were analysed using thematic analysis and team discussion to synthesise themes. Focus group participants identified the value of the VCPS to patients, to the health service and to themselves. They also identified enhancements to increase value for each of these groups. Perceived benefits to patients included access to specialist medication advice and improved medication knowledge. Staff valued access to an additional, trusted workforce who provided back-up and guidance. Staff also reported confidence in improved patient safety and identification of medication errors. Enhanced compliance with antimicrobial stewardship and hospital accreditation standards were beneficial to the health service. Suggested improvements included extending virtual service hours and widening patient eligibility to include aged care patients. The VCPS brought a positive, collegiate culture regarding medications. Healthcare staff perceived the VCPS was effective and an efficient way for the health service to supply pharmacy services to smaller hospitals. The ease of use, model of delivery, availability, local knowledge and responsiveness of highly skilled pharmacists was the key to user satisfaction. ANZCTR ACTRN12619001757101 , 11/12/2019.


Replacement of H1 linker histone during bovine somatic cell nuclear transfer.
Linker histone variants are involved in regulation of chromosome organization and gene transcription; several subtypes are expressed in the maturing oocyte and developing embryo. In Xenopus and mice, the transition between linker histone variants occurred following nuclear transfer, and apparently contributed to donor nuclear reprogramming. To determine whether such linker histone replacement occurred after bovine nuclear transfer, red fluorescent protein (RFP) tagged H1e (somatic linker histone H1e) donor cells and Venus tagged H1foo eggs were created, enucleated eggs were injected with donor cells, and embryos were created by fusion. Using fluorescence microscopy, release of H1e in the donor nucleus, acquisition of H1foo by donor chromosomes, and the H1foo-to-H1e transition were observed in live cells. Linker histone replacement occurred more slowly in bovine than murine embryos. Low levels of diffuse red fluorescence (H1e) in the donor nucleus were detected 5 h after fusion, at which time green fluorescence (H1foo) had incorporated into donor chromosomes. However, complete replacement did not occur until 8 h after fusion. We concluded that the linker histone transition was sufficiently conserved among species, which provided further evidence regarding its important role in nuclear reprogramming.


Detecting splicing patterns in genes involved in hereditary breast and ovarian cancer.
Interpretation of variants of unknown significance (VUS) is a major challenge for laboratories performing molecular diagnosis of hereditary breast and ovarian cancer (HBOC), especially considering that many genes are now known to be involved in this syndrome. One important way these VUS can have a functional impact is through their effects on RNA splicing. Here we present a custom RNA-Seq assay plus bioinformatics and biostatistics pipeline to analyse specifically alternative and abnormal splicing junctions in 11 targeted HBOC genes. Our pipeline identified 14 new alternative splices in BRCA1 and BRCA2 in addition to detecting the majority of known alternative spliced transcripts therein. We provide here the first global splicing pattern analysis for the other nine genes, which will enable a comprehensive interpretation of splicing defects caused by VUS in HBOC. Previously known splicing alterations were consistently detected, occasionally with a more complex splicing pattern than expected. We also found that splicing in the 11 genes is similar in blood and breast tissue, supporting the utility and simplicity of blood splicing assays. Our pipeline is ready to be integrated into standard molecular diagnosis for HBOC, but it could equally be adapted for an integrative analysis of any multigene disorder.


Intrathoracic versus Cervical ANastomosis after minimally invasive esophagectomy for esophageal cancer: study protocol of the ICAN randomized controlled trial.
Currently, a cervical esophagogastric anastomosis (CEA) is often performed after minimally invasive esophagectomy (MIE). However, the CEA is associated with a considerable incidence of anastomotic leakage requiring reintervention or reoperation and moderate functional results. An intrathoracic esophagogastric anastomosis (IEA) might reduce the incidence of anastomotic leakage, improve functional results and reduce costs. The objective of the ICAN trial is to compare anastomotic leakage and postoperative morbidity, mortality, quality of life and cost-effectiveness between CEA and IEA after MIE. The ICAN trial is an open randomized controlled multicentre superiority trial, comparing CEA (control group) with IEA (intervention group) after MIE. All patients with esophageal cancer planning to undergo curative MIE are considered for inclusion. A total of 200 patients will be included in the study and randomized between the groups in a 1:1 ratio. The primary outcome is anastomotic leakage requiring reintervention or reoperation, and secondary outcomes are (amongst others) other postoperative complications, new onset of organ failure, length of stay, mortality, benign strictures requiring dilatation, quality of life and cost-effectiveness. We hypothesize that an IEA after MIE is associated with a lower incidence of anastomotic leakage requiring reintervention or reoperation than a CEA. The trial is also designed to give answers to additional research questions regarding a possible difference in functional outcome, quality of life and cost-effectiveness. Netherlands Trial Register: NTR4333 . Registered on 23 December 2013.


The effects of TiO2 nanotube arrays with different diameters on macrophage/endothelial cell response and ex vivo hemocompatibility.
Percutaneous coronary intervention with stenting is the most widely adopted surgical technique for the treatment of coronary disease. However, in-stent restenosis (ISR) continues to cause serious concern. Improving the functionality of endothelial cells (ECs) is of importance for dealing with ISR. However, limited successes in in vivo studies imply that the underlying mechanism relating to the process of intervention with a material has not been well understood to date. Another determining factor that has long been underestimated in the design of stents is immune response dominated by macrophages (MΦs). In the present study, TiO2 nanotube arrays (TNAs) with different diameters were fabricated by anodization. The diameter could be precisely adjusted from 15 nm (Nano-15) to 120 nm (Nano-120). The effect of distinct nanodimensions on the behavior of ECs/MΦs and their crosstalk was investigated. The results indicated that Nano-15 not only promoted the initial vitality of ECs and function-related gene expression but also enhanced the activation and spread of MΦs. In addition, Nano-15 downregulated the gene expression of inflammatory cytokines and pro-inflammatory M1 markers, while upregulating the gene expression of pro-healing M2 markers, autophagy markers, and growth factors of MΦs. A further investigation conducted using a specialized specimen-conditioned medium (SCM) from MΦs/ECs disclosed that the SCM derived from Nano-15 was able to manipulate a favorable microenvironment to facilitate MΦ-EC crosstalk via downregulation of inflammation-related gene expression and upregulation of function-related gene expression in ECs. Moreover, we further investigated the hemocompatibility of specimens in an ex vivo animal model. The results indicated that both pure Ti and Nano-15 possess satisfactory hemocompatibility, as manifested by the few thrombi that formed around the specimens. Additionally, in comparison with Nano-120, Nano-15 can significantly alleviate platelet activation and manipulate a loose structure of fibrin fibers in a dynamic situation. This study suggests that the application of Nano-15 as a surface coating for coronary stents may be a promising strategy for preventing ISR.


Aleukemic leukemia presenting with paranoid psychosis.
Mast cells not only synthesize and release serotonin, but also express and are activated through multiple serotonin receptors. Low blood serotonin level might define a specific subset of patients with systemic mastocytosis (SM) who are more likely to present with neurologic and gastrointestinal complaints. Mast cell leukemia (MCL) is a rare and aggressive type of systemic mastocytosis, and psychiatric manifestations in its course have not been well-characterized. We describe herein a unique patient with a KIT D816V mutation positive a leukemic leukemia variant of systemic mastocytosis with gastrointestinal involvement, presenting with a severe and sustained paranoid delusional illness. While diarrhea improved with the use of histamine H1 and H2 receptor antagonists, the psychosis did not, and the disease followed a dramatic course with a rapidly fatal outcome. As there is paucity of literature, diagnosis of MCL presenting with psychiatric symptoms remains a diagnostic challenge and warrants clinicians to be alert of this rare possibility.


Non-parametric and semi-parametric support estimation using SEquential RESampling random walks on biomolecular sequences.
Non-parametric and semi-parametric resampling procedures are widely used to perform support estimation in computational biology and bioinformatics. Among the most widely used methods in this class is the standard bootstrap method, which consists of random sampling with replacement. While not requiring assumptions about any particular parametric model for resampling purposes, the bootstrap and related techniques assume that sites are independent and identically distributed (i.i.d.). The i.i.d. assumption can be an over-simplification for many problems in computational biology and bioinformatics. In particular, sequential dependence within biomolecular sequences is often an essential biological feature due to biochemical function, evolutionary processes such as recombination, and other factors. To relax the simplifying i.i.d. assumption, we propose a new non-parametric/semi-parametric sequential resampling technique that generalizes "Heads-or-Tails" mirrored inputs, a simple but clever technique due to Landan and Graur. The generalized procedure takes the form of random walks along either aligned or unaligned biomolecular sequences. We refer to our new method as the SERES (or "SEquential RESampling") method. To demonstrate the performance of the new technique, we apply SERES to estimate support for the multiple sequence alignment problem. Using simulated and empirical data, we show that SERES-based support estimation yields comparable or typically better performance compared to state-of-the-art methods.