bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunohistochemical loss of 5‐hydroxymethylcytosine expression in acute myeloid leukaemia: relationship to somatic gene mutations affecting epigenetic pathways
Authors: Magotra Minoti , Sakhdari Ali , Lee Paul J , Tomaszewicz Keith , Dresser Karen , Hutchinson Lloyd M , Woda Bruce A , Chen Benjamin J
Published: July 2016

Aims

Genes affecting epigenetic pathways are frequently mutated in myeloid malignancies, including acute myeloid leukaemia (AML). The genes encoding TET2, IDH1 and IDH2 are among the most commonly mutated genes, and cause defective conversion of 5‐methylcytosine into 5‐hydroxymethylcytosine (5hmC), impairing demethylation of DNA, and presumably serving as driver mutations in leukaemogenesis. The aim of this study was to correlate 5hmC immunohistochemical loss with the mutation status of genes involved in epigenetic pathways in AML.

Methods and results

Immunohistochemical staining with an anti‐5hmC antibody was performed on 41 decalcified, formalin‐fixed paraffin‐embedded (FFPE) bone marrow biopsies from patients with AML. Archived DNA was subjected to next‐generation sequencing for analysis of a panel of genes, including TET2, IDH1, IDH2, WT1 and DNMT3A. TET2, IDH1, IDH2, WT1 and DNMT3A mutations were found in 46% (19/41) of the cases. Ten of 15 cases (67%) with TET2, IDH1, IDH2 or WT1 mutations showed deficient 5hmC staining, whereas nine of 26 cases (35%) without a mutation in these genes showed loss of 5hmC. It is of note that all four cases with TET2 mutations showed deficient 5hmC staining.

Conclusions

Overall, somatic mutations in TET2, IDH1, IDH2, WT1 and DNMT3A were common in our cohort of AML cases. Immunohistochemical staining for 5hmC was lost in the majority of cases harbouring mutations in these genes, reflecting the proposed relationship between dysfunctional epigenetic pathways and leukaemogenesis.
Metabonomics: a developing platform for better understanding Chinese herbal teas as a complementary therapy
Authors: You Rong , Guan Yanqing , Li Lin
Published: July 2016

Chinese herbal teas (CHTs) have gained increasing attention throughout the world as a complementary therapy. Due to the multicomponent composition in herbal teas, the molecular mechanisms of their causality in health effects are still elusive and becoming the main obstacles for their application. There is a pressing need for a more systematic study about their molecular mechanisms to substantiate the health effects of CHTs. With the particular feature of measuring the real metabolic phenotype resulted from molecular interplay between host and environmental factors, metabonomics is especially suited to inspect metabolic responses to CHT intake and provide insights into health effects. Application of metabonomics to uncover the effects of CHTs on human health is foreseen as providing deeper insights of the complicated molecular mechanisms of CHTs by explaining relatively subtle and different effects of herbal tea intervention, classifying the metabolic responses to herbal tea, depicting synergistic effects of CHTs and evaluating the safety of CHTs intake.
Down Syndrome and Fragile X Syndrome in a Colombian Woman: Case Report
Authors: Saldarriaga Wilmar , Ruiz Fabian Andres , Tassone Flora , Hagerman Randi
Published: July 2016

Background

Down syndrome (DS) and Fragile X syndrome (FXS) are the major genetic causes of intellectual disabilities. Here, we present a case of a 32‐year‐old woman with the diagnosis of both FXS and DS. She is the daughter of a 47‐year‐old pre‐mutation woman who also has three sons with FXS.

Methods

Cytogenetic testing detected the presence of a complete trisomy 21. A combination of PCR and Southern blot analysis was utilized to document the presence of the FMR1 full mutation.

Results

The patient has physical characteristics and behavioural disturbances typical of both FXS and DS, which were confirmed by molecular testing. Her treatment plan included a trial of sertraline because of the severity of her shyness and lack of language. She had an excellent response to sertraline with improvement in shyness and social interactions, particularly with family members.

Conclusions

In this study, we report the case of a woman with both FXS and DS, which is the fifth case of FXS and DS in the world's literature. The patient is from Ricaurte, a small town in Colombia, South America, where there is the world's highest prevalence for FXS.
Training of Residential Social Care Staff to Meet the Needs of Older People with Intellectual Disabilities who Develop Age‐Related Health Problems: An Exploratory Study
Authors: Northway Ruth , Jenkins Robert , Holland‐Hart Daniella
Published: July 2016

Background

Despite awareness of the age related health needs of people with intellectual disabilities little is known regarding how residential social care staff are prepared to meet such needs.

Methods

Data were gathered via semi‐structured interviews from 14 managers of supported living settings. Transcripts were thematically analysed.

Findings

Staff may work in supported living settings with no prior experience of care work, and previous knowledge/experience of supporting people in relation to their health is not required. Whilst health related training is provided there is a lack of specific training regarding healthy ageing, and training seems to be reactive to changing needs of tenants meaning that proactive monitoring for changes in health status may not occur.

Conclusions

Whilst some training is provided for residential social care staff in relation to health and ageing a more proactive approach is required which should include a focus on healthy ageing.
Effectiveness of a Communication for Behavioral Impact (COMBI) Intervention to Reduce Salt Intake in a Vietnamese Province Based on Estimations From Spot Urine Samples
Authors: Do Ha Thi Phuong , Santos Joseph Alvin , Trieu Kathy , Petersen Kristina , Le Mai Bach , Lai Duc Truong , Bauman Adrian , Webster Jacqui
Published: July 2016

This study evaluated the effectiveness of the Communication for Behavioral Impact (COMBI)–Eat Less Salt intervention conducted in Viet Tri, Vietnam. The behavior change intervention was implemented in four wards and four communes for one year, which included mass media communication, school interventions, community programs, and focus on high‐risk groups. Mean sodium excretion was estimated from spot urine samples using different equations. A subsample provided 24‐hour urine to validate estimates from spot urine. Information about salt‐related knowledge and behaviors was also collected. There were 513 participants at both baseline and follow‐up. Mean sodium excretion estimated from spot urines fell significantly from 8.48 g/d at baseline to 8.05 g/d at follow‐up (P=.001). All spot equations demonstrated a significant reduction in sodium levels; however, the change was smaller than the measured 24‐hour urine. Participants showed improved knowledge and behaviors following the intervention. The COMBI intervention was effective in lowering average population salt intake and improving knowledge and behaviors.
Effectiveness of a Proactive Primary Care Program on Preserving Daily Functioning of Older People: A Cluster Randomized Controlled Trial
Authors: Bleijenberg Nienke , Drubbel Irene , Schuurmans Marieke J. , Dam Hester ten , Zuithoff Nicolaas P.A. , Numans Mattijs E. , de Wit Niek J.
Published: July 2016

Objectives

To determine the effectiveness of a proactive primary care program on the daily functioning of older people in primary care.

Design

Single‐blind, three‐arm, cluster‐randomized controlled trial with 1‐year follow‐up.

Setting

Primary care setting, 39 general practices in the Netherlands.

Participants

Community‐dwelling people aged 60 and older (N = 3,092).

Interventions

A frailty screening intervention using routine electronic medical record data to identify older people at risk of adverse events followed by usual care from a general practitioner; after the screening intervention, a nurse‐led care program consisting of a comprehensive geriatric assessment, evidence‐based care planning, care coordination, and follow‐up; usual care.

Measurements

Primary outcome was daily functioning measured using the Katz‐15 (6 activities of daily living (ADLs), 8 instrumental activities of daily living (IADLs), one mobility item (range 0–15)); higher scores indicate greater dependence. Secondary outcomes included quality of life, primary care consultations, hospital admissions, emergency department visits, nursing home admissions, and mortality.

Results

The participants in both intervention arms had less decline in daily functioning than those in the usual care arm at 12 months (mean Katz‐15 score: screening arm, 1.87, 95% confidence interval (CI) = 1.77–1.97; screening and nurse‐led care arm, 1.88, 95% CI = 1.80–1.96; control group, 2.03, 95% CI = 1.92–2.13; P = .03). No differences in quality of life were observed.

Conclusion

Participants in both intervention groups had less decline than those in the control group at 1‐year follow‐up. Despite the statistically significant effect, the clinical relevance is uncertain at this point because of the small differences. Greater customizing of the intervention combined with prolonged follow‐up may lead to more‐robust results.
Oral lichen planus and epithelial dysplasia with lichenoid features: a review and discussion with special reference to diagnosis
Authors: Sanketh D. SharathKumar , Patil Shankargouda , Swetha Balamukundan
Published: July 2016

Despite guidelines for the diagnosis of oral lichen planus (OLP), there has been much difficulty and ambiguity in rendering a straightforward diagnosis. The root of the problem might arise in not following universal guidelines and being too rigid and unilateral in making a diagnosis. Because of its autoimmune pathogenesis, the dynamic nature of OLP has further fueled confusions. This has had repercussions in the form of controversies in its diagnosis, treatment protocols, research, and most importantly, its potentially malignant nature. Oral lichenoid dysplasia (OLD), an enigmatic entity, proposed by Krutchkoff and Eisenberg, has not found universal acceptance by the pathology community. The objective of the present review was to outline these debates and discuss ongoing controversies regarding OLD and uncertainties in the diagnostic criteria of OLP.
Strange and scary memories of the intensive care unit: a qualitative, longitudinal study inspired by Ricoeur's interpretation theory
Authors: Svenningsen Helle , Egerod Ingrid , Dreyer Pia
Published: July 2016

Aims and objectives

To describe the content of former intensive care unit patients’ memories of delusions.

Background

Intensive care unit patients often have strange and frightening experiences during the critical stage of illness. Earlier studies have provided small‐sample in‐depth descriptions of patient experiences in intensive care unit, but large‐scale studies are also needed to inform intensive care unit follow‐up.

Methods

The study had a qualitative design using phenomenological hermeneutic analysis inspired by Ricoeur's interpretive theory. Patients were assessed with Confusion Assessment Method of the Intensive Care Unit for delirium in intensive care unit, and after discharge, memories of delusions were described by 114 of 325 patients in face‐to‐face (after two weeks) and telephone interviews (after two and six months) using the Intensive Care Unit Memory Tool.

Results

Four themes emerged: the ever‐present family, dynamic spaces, surviving challenges and constant motion. Memories of delusions were a vivid mix of fact and fiction, demonstrating dynamic shifts in time, place and motion, but not dependent on the presence of delirium assessed by Confusion Assessment Method of the Intensive Care Unit.

Conclusions

Analysis based on Ricoeurian phenomenological hermeneutics provided insights into themes in intensive care unit patients’ memories of delusions. More studies are needed to understand the meaning of memories of delusions, the commonality of themes and the association between delusions and delirium after an intensive care unit stay.

Relevance to clinical practice

Understanding patients’ memories of delusions is beneficial to nurses caring for patients that are anxious, upset or agitated. It opens a window to the world of the patient who is unable to communicate due to intubation and general weakness. We recommend the provision of nurse‐led intensive care unit follow‐up enabling patients to describe and discuss their intensive care unit experiences.
A MIXED CONTROL PROBLEM OF THE MANAGEMENT OF NATURAL RESOURCES
Authors: ALVAREZ MARÍA EVANGELINA , DI MARCO SILVIA , ERDLENBRUCH KATRIN , TIDBALL MABEL
Published: July 2016

In this paper, we study the optimal solutions of a model of natural resource management which allows for both impulse and continuous harvesting policies. This type of model is known in the literature as mixed optimal control problem. In the resource management context, each type of control represents a different harvesting technology, which has a different cost. In particular, we want to know when the following conjecture made by Clark is an optimal solution to this mixed optimal control problem: if the harvesting capacity is unlimited, it is optimal to jump immediately to the steady state of the continuous time problem and then to stay there. We show that under a particular relationship between the continuous and the impulse profit function, the conjecture made by Clark is true. In other cases, however, it is either better to use only continuous control variables or to jump to resource levels which are smaller than the steady state and then let the resource grow back to the steady state. These results emphasize the importance of the cost functions in the modeling of natural resource management.
Geographical thinking in nursing inquiry, part two: performance, possibility, and non‐representational theory
Authors: Andrews Gavin J.
Published: July 2016

Part one in this two paper series reviewed the nature of geographical thinking in nursing research thus far. The current paper builds on it by looking forwards and providing a particular vision for future research. It argues that it is time to once again look to the parent discipline of human geography for inspiration, specifically to its turn towards non‐representational theory, involving an emphasis on life that onflows prior to meaning, significance, and full cognition; on life's ‘taking‐place’. The paper introduces this way of viewing and animating the world. Some potential connections to nursing research and practice are suggested, as are some specific avenues for future inquiry. Explained is how, through non‐representational theory, nursing might be re‐imagined as something that reveals space‐time.
Evolution of Clinical and Electrophysiological Data in Children with a Preexcitation Syndrome
Authors: BREMBILLA‐PERROT BÉATRICE , SELLAL JEAN‐MARC , OLIVIER ARNAUD , VILLEMIN THIBAUT , MOULIN‐ZINSCH ANNE , BEURRIER DANIEL , LETHOR JEAN‐PAUL , MARÇON FRANÇOIS , DE CHILLOU CHRISTIAN , FELBLINGER JACQUES , VINCENT JULIE
Published: July 2016

Background

With ablation, the follow‐up of preexcitation syndrome now is difficult to assess. The purpose was to collect data of children with a preexcitation syndrome studied on two separate occasions within a minimal interval of 1 year.

Methods

This is a retrospective chart review of 47 children initially aged 12 ± 4 years, who underwent two or more invasive electrophysiological studies (EPS) within 1–25 years of one another (6.3 ± 4.8) for occurrence of symptoms or new evaluation.

Results

Among initially symptomatic children (n = 25), four (19%) became asymptomatic and one presented life‐threatening arrhythmia. Among asymptomatic children (n = 22), five became symptomatic (22.7%). Anterograde conduction disappeared in seven of 23 children with initially long accessory pathway‐effective refractory period, but four of six had still induced atrioventricular reentrant tachycardia (AVRT). AVRT was induced at second EPS in three of 13 asymptomatic preexcitation syndrome with negative initial EPS. There were no spontaneous adverse events in the five children with criteria of malignancy at initial EPS; signs of malignancy disappeared in two. At multivariate analysis, AVRT at initial EPS was the only independent factor of symptomatic AVRT during follow‐up. Absence of induced AVRT at initial EPS was the only factor of absence of symptoms and a negative study at the second EPS.

Conclusions

There were no significant changes of data in children after 6.3 ± 4.8 years of follow‐up. Most children with spontaneous/inducible AVRTs at initial EPS had still inducible AVRT at second EPS. Induced AF conducted with high rate has a relatively low prognostic value for the prediction of adverse events.
Atheism and Morality, Guilt and Shame: Why the Moral Complacency of the New Atheism is a Mistake
Authors: Lynch Tony , Dahanayake Nishanathe
Published: July 2016

When it comes to morality, the New Atheists appear to think that their rejection of religion, except for the removal of fundamentalist distortions, changes nothing. We think that this is because they have not thought things through. Atheism might not be a threat to shame morality, but it is certainly a threat to guilt morality. Given that there are reasons to doubt the viability today of shame morality, we face a far greater problem if atheism triumphs than the New Atheists admit.
Clinical outcomes of Ghanaian Buruli ulcer patients who defaulted from antimicrobial therapy
Authors: Klis S. , Kingma R. A. , Tuah W. , van der Werf T. S. , Stienstra Y.
Published: July 2016

Objectives

Buruli ulcer (BU) is a tropical skin disease caused by infection with Mycobacterium ulcerans, which is currently treated with 8 weeks of streptomycin and rifampicin. The evidence to treat BU for a duration of 8 weeks is limited; a recent retrospective study from Australia suggested that a shorter course of antimicrobial therapy might be equally effective. We studied the outcomes of BU in a cohort of Ghanaian patients who defaulted from treatment and as such received less than 8 weeks of antimicrobial therapy.

Methods

A number of days of antimicrobial therapy and patient and lesion characteristics were recorded from charts from a cohort of BU patients treated at Nkawie‐Toase hospital between 2008 and 2012. Patients who defaulted from treatment were retrieved, and lesion characteristics and functional limitations were recorded.

Results

About 54% of patients defaulted from therapy or wound care. Forty‐seven defaulters with follow‐up completed had received <56 days of antibiotics. 84% of these patients healed after 32 days or less of antibiotics. There appeared to be an increased rate of healing in smaller lesions; 94% of WHO category I lesions had healed after 32 days or less of antibiotics.

Conclusion

Although numbers were small, and a potential for bias exists, our findings suggest that a reduction in the duration of antimicrobial therapy in BU in small, early lesions is feasible. These findings can serve as a basis for future well‐designed studies.
Exercise training preserves vagal preganglionic neurones and restores parasympathetic tonus in heart failure
Authors: Ichige Marcelo H. A. , Santos Carla R. , Jordão Camila P. , Ceroni Alexandre , Negrão Carlos E. , Michelini Lisete C.
Published: July 2016

Key points

Heart Failure (HF) is accompanied by reduced ventricular function, activation of compensatory neurohormonal mechanisms and marked autonomic dysfunction characterized by exaggerated sympathoexcitation and reduced parasympathetic activity.

With 6 weeks of exercise training, HF‐related loss of choline acetyltransferase (ChAT)‐positive vagal preganglionic neurones is avoided, restoring the parasympathetic tonus to the heart, and the immunoreactivity of dopamine β‐hydroxylase‐positive premotor neurones that drive sympathetic outflow to the heart is reduced. Training‐induced correction of autonomic dysfunction occurs even with the persistence of abnormal ventricular function.

Strong positive correlation between improved parasympathetic tonus to the heart and increased ChAT immunoreactivity in vagal preganglionic neurones after training indicates this is a crucial mechanism to restore autonomic function in heart failure.

Exercise training is an efficient tool to attenuate sympathoexcitation, a hallmark of heart failure (HF). Although sympathetic modulation in HF is widely studied, information regarding parasympathetic control is lacking. We examined the combined effects of sympathetic and vagal tonus to the heart in sedentary (Sed) and exercise trained (ET) HF rats and the contribution of respective premotor and preganglionic neurones. Wistar rats submitted to coronary artery ligation or sham surgery were assigned to training or sedentary protocols for 6 weeks. After haemodynamic, autonomic tonus (atropine and atenolol i.v.) and ventricular function determinations, brains were collected for immunoreactivity assays (choline acetyltransferase, ChATir; dopamine β‐hydroxylase, DBHir) and neuronal counting in the dorsal motor nucleus of vagus (DMV), nucleus ambiguus (NA) and rostroventrolateral medulla (RVLM). HF‐Sed vs. SHAM‐Sed exhibited decreased exercise capacity, reduced ejection fraction, increased left ventricle end diastolic pressure, smaller positive and negative dP/dt, decreased intrinsic heart rate (IHR), lower parasympathetic and higher sympathetic tonus, reduced preganglionic vagal neurones and ChATir in the DMV/NA, and increased RVLM DBHir. Training increased treadmill performance, normalized autonomic tonus and IHR, restored the number of DMV and NA neurones and corrected ChATir without affecting ventricular function. There were strong positive correlations between parasympathetic tonus and ChATir in NA and DMV. RVLM DBHir was also normalized by training, but there was no change in neurone number and no correlation with sympathetic tonus. Training‐induced preservation of preganglionic vagal neurones is crucial to normalize parasympathetic activity and restore autonomic balance to the heart even in the persistence of cardiac dysfunction.
Guidance and Methodologies for Managing Digital Instrumentation and Control Obsolescence
Authors: O’Connor Matthew , Geddes Bruce , Kelley Sean
Published: July 2016

New nuclear plant technology will rely heavily, if not exclusively, on digital equipment. Obsolescence of digital instrumentation and control (I&C) equipment is an inevitable part of plant technology life cycle for new and existing plants. Developing an overall strategic plan can mitigate some of the risks associated with obsolescence. Moreover, when developed as a part of an overall life cycle management plan (LCMP), a strategic obsolescence management approach can identify steps that can be taken at early stages of the technology life cycle to cope proactively with the obsolescence of equipment. Recent work within the advanced nuclear technology (ANT) program at the Electric Power Research Institute (EPRI) (O’Connor et al., 2014, “Advanced Nuclear Technology: Guidance and Methodologies for Managing Digital Instrumentation and Control Obsolescence,” EPRI, Palo Alto, CA, Product ID 3002002852) has developed guidance and methodologies for determining when digital obsolescence is likely to occur, the extent to which it can occur, the risks and impacts due to obsolescence, and strategies that can be used to minimize its effects, all in the context of system LCMP. Worksheets for assessing obsolescence risks and the applicability and limitations of management strategies were developed as a part of this work, and can be used to create or supplement a strategic obsolescence management plan.
Homogenization and Path Independence of the J-Integral in Heterogeneous Materials
Authors: Hsueh Chun-Jen , Bhattacharya Kaushik
Published: July 2016

The J-integral that determines the driving force on a crack tip is a central concept of fracture mechanics. It is particularly useful since it is path independent in homogeneous materials. However, most materials are heterogeneous at a microscopic scale, and the J-integral is not necessarily path independent in heterogeneous media. In this paper, we prove the existence of an effective J-integral in heterogeneous media, show that it can be computed from the knowledge of the macroscopic or homogenized displacement fields and that it is path independent in macroscopically homogeneous media as long as the contours are large compared to the length scale of the heterogeneities. This result justifies the common engineering use of the J-integral.
Physics-Based Multivariable Modeling and Feedback Linearization Control of Melt-Pool Geometry and Temperature in Directed Energy Deposition
Authors: Wang Qian , Li Jianyi , Gouge Michael , Nassar Abdalla R. , (Pan) Michaleris Panagiotis , Reutzel Edward W.
Published: July 2016

There has been continuing effort in developing analytical, numerical, and empirical models of laser-based additive manufacturing (AM) processes in the literature. However, advanced physics-based models that can be directly used for feedback control design, i.e., control-oriented models, are severely lacking. In this paper, we develop a physics-based multivariable model for directed energy deposition. One important difference between our model from the existing work lies in a novel parameterization of the material transfer rate in the deposition as a function of the process operating parameters. Such parameterization allows an improved characterization of the steady-state melt-pool geometry compared to the existing lumped-parameter models. Predictions of melt-pool geometry and temperature from our model are validated using experimental data obtained from deposition of Ti-6AL-4V and deposition of Inconel® 718 on a laser engineering net shaping (LENS) AM process and finite-element analysis. Then based on this multivariable model, we design a nonlinear multi-input multi-output (MIMO) control, specifically a feedback linearization (FL) control, to track both melt-pool height and temperature reference trajectories using laser power and laser scan speed.
The Effects of Helmet Weight on Hybrid III Head and Neck Responses by Comparing Unhelmeted and Helmeted Impacts
Authors: Jadischke Ron , Viano David C. , McCarthy Joe , King Albert I.
Published: July 2016

Most studies on football helmet performance focus on lowering head acceleration-related parameters to reduce concussions. This has resulted in an increase in helmet size and mass. The objective of this paper was to study the effect of helmet mass on head and upper neck responses. Two independent test series were conducted. In test series one, 90 pendulum impact tests were conducted with four different headform and helmet conditions: unhelmeted Hybrid III headform, Hybrid III headform with a football helmet shell, Hybrid III headform with helmet shell and facemask, and Hybrid III headform with the helmet and facemask with mass added to the shell (n = 90). The Hybrid III neck was used for all the conditions. For all the configurations combined, the shell only, shell and facemask, and weighted helmet conditions resulted in 36%, 43%, and 44% lower resultant head accelerations (p < 0.0001), respectively, when compared to the unhelmeted condition. Head delta-V reductions were 1.1%, 4.5%, and 4.4%, respectively. In contrast, the helmeted conditions resulted in 26%, 41%, and 49% higher resultant neck forces (p < 0.0001), respectively. The increased neck forces were dominated by neck tension. In test series two, testing was conducted with a pneumatic linear impactor (n = 178). Fourteen different helmet makes and models illustrate the same trend. The increased neck forces provide a possible explanation as to why there has not been a corresponding reduction in concussion rates despite improvements in helmets ability to reduce head accelerations.
Basidiomycete yeasts in the cortex of ascomycete macrolichens
Authors: Spribille Toby , Tuovinen Veera , Resl Philipp , Vanderpool Dan , Wolinski Heimo , Aime M. Catherine , Schneider Kevin , Stabentheiner Edith , Toome-Heller Merje , Thor Göran , Mayrhofer Helmut , Johannesson Hanna , McCutcheon John P.
Published: July 2016

Lichens assemble in three parts

Lichen growth forms cannot be recapitulated in the laboratory by culturing the plant and fungal partners together. Spribille
et al.
have discovered that the classical binary view of lichens is too simple. Instead, North American beard-like lichens are constituted of not two but three symbiotic partners: an ascomycetous fungus, a photosynthetic alga, and, unexpectedly, a basidiomycetous yeast. The yeast cells form the characteristic cortex of the lichen thallus and may be important for its shape. The yeasts are ubiquitous and essential partners for most lichens and not the result of lichens being colonized or parasitized by other organisms.

Science
, this issue p.
488
Three risk factors, one mechanism
Authors: Ferrarelli Leslie K.
Published: July 2016

Three mutations associated with Alzheimer’s disease are linked in the clearance of β-amyloid.
How sweet it isn’t
Authors: Wong Wei
Published: July 2016

Synthetic sweeteners trigger a neuronal circuit that stimulates appetite and causes sweet taste sensitization.
Development of a Novel High-Density [
3
H]Hypoxanthine Scintillation Proximity Assay To Assess Plasmodium falciparum Growth
Authors: de Cózar Cristina , Caballero Iván , Colmenarejo Gonzalo , Sanz Laura M. , Álvarez-Ruiz Emilio , Gamo Francisco-Javier , Cid Concepción
Published: July 2016

The discovery and development of new antimalarial drugs are becoming imperative because of the spread of resistance to current clinical treatments. The lack of robustly validated antimalarial targets and the difficulties with the building in of whole-cell activity in screening hits are hampering target-based approaches. However, phenotypic screens of structurally diverse molecule libraries are offering new opportunities for the identification of novel antimalarials. Several methodologies can be used to determine the whole-cell
in vitro
potencies of antimalarial hits. The [
3
H]hypoxanthine incorporation assay is considered the “gold standard” assay for measurement of the activity of antimalarial compounds against intraerythrocytic forms of
Plasmodium falciparum
. However, the method has important limitations, as the assay is not amenable for high-throughput screening since it remains associated with the 96-well plate format. We have overcome this drawback by adapting the [
3
H]hypoxanthine incorporation method to a 384-well high-density format by coupling a homogeneous scintillation proximity assay (SPA) and thus eliminating the limiting filtration step. This SPA has been validated using a diverse set of 1,000 molecules, including both a representative set from the Tres Cantos Antimalarial Set (TCAMS) of compounds and molecules inactive against whole cells. The results were compared with those from the
P. falciparum
lactate dehydrogenase whole-cell assay, another method that is well established as a surrogate for parasite growth and is amenable for high-throughput screening. The results obtained demonstrate that the SPA-based [
3
H]hypoxanthine incorporation assay is a suitable design that is adaptable to high-throughput antimalarial drug screening and that maintains the features, robustness, and reliability of the standard filtration hypoxanthine incorporation method.
Inhibition of Leishmania mexicana Growth by the Tuberculosis Drug SQ109
Authors: García-García Verónica , Oldfield Eric , Benaim Gustavo
Published: July 2016

We report that the tuberculosis drug SQ109 [
N
-adamantan-2-yl-
N
′-((
E
)-3,7-dimethyl-octa-2,6-dienyl)-ethane-1,2-diamine] has potent activity against the intracellular amastigote form of
Leishmania mexicana
(50% inhibitory concentration [IC
50
], ∼11 nM), with a good selectivity index (>500). It is also active against promastigotes (IC
50
, ∼500 nM) and acts as a protonophore uncoupler, in addition to disrupting Ca
2+
homeostasis by releasing organelle Ca
2+
into the cytoplasm, and as such, it is an interesting new leishmaniasis drug hit candidate.
Short Palate, Lung, and Nasal Epithelial Clone 1 Has Antimicrobial and Antibiofilm Activities against the Burkholderia cepacia Complex
Authors: Ahmad Saira , Tyrrell Jean , Walton William G. , Tripathy Ashutosh , Redinbo Matthew R. , Tarran Robert
Published: July 2016

The opportunistic bacteria of the
Burkholderia cepacia
complex (Bcc) are extremely pathogenic to cystic fibrosis (CF) patients, and acquisition of Bcc bacteria is associated with a significant increase in mortality. Treatment of Bcc infections is difficult because the bacteria are multidrug resistant and able to survive in biofilms. Short palate, lung, and nasal epithelial clone 1 (SPLUNC1) is an innate defense protein that is secreted by the upper airways and pharynx. While SPLUNC1 is known to have antimicrobial functions, its effects on Bcc strains are unclear. We therefore tested the hypothesis that SPLUNC1 is able to impair Bcc growth and biofilm formation. We found that SPLUNC1 exerted bacteriostatic effects against several Bcc clinical isolates, including
B. cenocepacia
strain J2315 (50% inhibitory concentration [IC
50
] = 0.28 μM), and reduced biofilm formation and attachment (IC
50
= 0.11 μM). We then determined which domains of SPLUNC1 are responsible for its antimicrobial activity. Deletions of SPLUNC1's N terminus and α6 helix did not affect its function. However, deletion of the α4 helix attenuated antimicrobial activity, while the corresponding α4 peptide displayed antimicrobial activity. Chronic neutrophilia is a hallmark of CF lung disease, and neutrophil elastase (NE) cleaves SPLUNC1. However, we found that the ability of SPLUNC1 to disrupt biofilm formation was significantly potentiated by NE pretreatment. While the impact of CF on SPLUNC1-Bcc interactions is not currently known, our data suggest that understanding this interaction may have important implications for CF lung disease.
Divergent Transcriptional Responses to Physiological and Xenobiotic Stress in Giardia duodenalis
Authors: Ansell Brendan R. E. , McConville Malcolm J. , Baker Louise , Korhonen Pasi K. , Emery Samantha J. , Svärd Staffan G. , Gasser Robin B. , Jex Aaron R.
Published: July 2016

Understanding how parasites respond to stress can help to identify essential biological processes.
Giardia duodenalis
is a parasitic protist that infects the human gastrointestinal tract and causes 200 to 300 million cases of diarrhea annually. Metronidazole, a major antigiardial drug, is thought to cause oxidative damage within the infective trophozoite form. However, treatment efficacy is suboptimal, due partly to metronidazole-resistant infections. To elucidate conserved and stress-specific responses, we calibrated sublethal metronidazole, hydrogen peroxide, and thermal stresses to exert approximately equal pressure on trophozoite growth and compared transcriptional responses after 24 h of exposure. We identified 252 genes that were differentially transcribed in response to all three stressors, including glycolytic and DNA repair enzymes, a mitogen-activated protein (MAP) kinase, high-cysteine membrane proteins, flavin adenine dinucleotide (FAD) synthetase, and histone modification enzymes. Transcriptional responses appeared to diverge according to physiological or xenobiotic stress. Downregulation of the antioxidant system and α-giardins was observed only under metronidazole-induced stress, whereas upregulation of GARP-like transcription factors and their subordinate genes was observed in response to hydrogen peroxide and thermal stressors. Limited evidence was found in support of stress-specific response elements upstream of differentially transcribed genes; however, antisense derepression and differential regulation of RNA interference machinery suggest multiple epigenetic mechanisms of transcriptional control.
Complete Genome Sequences of Three Outbreak-Associated
Legionella pneumophila
Isolates
Authors: Morrison Shatavia S. , Desai Heta P. , Mercante Jeffrey W. , Lapierre Pascal , Raphael Brian H. , Musser Kimberlee , Winchell Jonas M.
Published: July 2016

We report here the complete genome sequences of three
Legionella pneumophila
isolates that are associated with a Legionnaires' disease outbreak in New York in 2012. Two clinical isolates (D7630 and D7632) and one environmental isolate (D7631) were recovered from this outbreak. A single isolate-specific virulence gene was found in D7632. These isolates were included in a large study evaluating the genomic resolution of various bioinformatics approaches for
L. pneumophila
serogroup 1 isolates.
Role of Serum Amyloid A, Granulocyte-Macrophage Colony-Stimulating Factor, and Bone Marrow Granulocyte-Monocyte Precursor Expansion in Segmented Filamentous Bacterium-Mediated Protection from Entamoeba histolytica
Authors: Burgess Stacey L. , Saleh Mahmoud , Cowardin Carrie A. , Buonomo Erica , Noor Zannatun , Watanabe Koji , Abhyankar Mayuresh , Lajoie Stephane , Wills-Karp Marsha , Petri William A.
Published: July 2016

Intestinal segmented filamentous bacteria (SFB) protect from ameba infection, and protection is transferable with bone marrow dendritic cells (BMDCs). SFB cause an increase in serum amyloid A (SAA), suggesting that SAA might mediate SFB's effects on BMDCs. Here we further explored the role of bone marrow in SFB-mediated protection. Transient gut colonization with SFB or SAA administration alone transiently increased the H3K27 histone demethylase Jmjd3, persistently increased bone marrow
Csf2ra
expression and granulocyte monocyte precursors (GMPs), and protected from ameba infection. Pharmacologic inhibition of Jmjd3 H3K27 demethylase activity during SAA treatment or blockade of granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling in SFB-colonized mice prevented GMP expansion, decreased gut neutrophils, and blocked protection from ameba infection. These results indicate that alteration of the microbiota and systemic exposure to SAA can influence myelopoiesis and susceptibility to amebiasis via epigenetic mechanisms. Gut microbiota-marrow communication is a previously unrecognized mechanism of innate protection from infection.
The SaeRS Two-Component System Is a Direct and Dominant Transcriptional Activator of Toxic Shock Syndrome Toxin 1 in Staphylococcus aureus
Authors: Baroja Miren L. , Herfst Christine A. , Kasper Katherine J. , Xu Stacey X. , Gillett Daniel A. , Li Jingru , Reid Gregor , McCormick John K.
Published: July 2016

Toxic shock syndrome toxin 1 (TSST-1) is a
Staphylococcus aureus
superantigen that has been implicated in both menstrual and nonmenstrual toxic shock syndrome (TSS). Despite the important role of TSST-1 in severe human disease, a comprehensive understanding of staphylococcal regulatory factors that control TSST-1 expression remains incomplete. The
S. aureus
exotoxin expression (Sae) operon contains a well-characterized two-component system that regulates a number of important exotoxins in
S. aureus
, although regulation of TSST-1 by the Sae system has not been investigated. We generated a defined deletion mutant of the Sae histidine kinase sensor (
saeS
) in the prototypic menstrual TSS strain
S. aureus
MN8. Mutation of
saeS
resulted in a complete loss of TSST-1 expression. Using both luciferase reporter experiments and quantitative real-time PCR, we demonstrate that the Sae system is an important transcriptional activator of TSST-1 expression. Recombinant SaeR was able to bind directly to the
tst
promoter to a region containing two SaeR consensus binding sites. Although the stand-alone SarA transcriptional regulator has been shown to be both a positive and a negative regulator of TSST-1, deletion of
sarA
in
S. aureus
MN8 resulted in a dramatic overexpression of TSST-1. As expected, mutation of
agr
also reduced TSST-1 expression, but this phenotype appeared to be independent of Sae. A double mutation of
saeS
and
sarA
resulted in the loss of TSST-1 expression. This work indicates that the Sae system is a dominant and direct transcriptional activator that is required for expression of TSST-1.

IMPORTANCE
The TSST-1 superantigen is an exotoxin, produced by some strains of
S. aureus
, that has a clear role in both menstrual and nonmenstrual TSS. Although the well-characterized
agr
quorum sensing system is a known positive regulator of TSST-1, the molecular mechanisms that directly control TSST-1 expression are only partially understood. Our studies demonstrate that the Sae two-component regulatory system is a positive transcriptional regulator that binds directly to the TSST-1 promoter, and furthermore, our data suggest that Sae is required for expression of TSST-1. This work highlights how major regulatory circuits can converge to fine-tune exotoxin expression and suggests that the Sae regulatory system may be an important target for antivirulence strategies.
Complementation of Cobalamin Auxotrophy in Synechococcus sp. Strain PCC 7002 and Validation of a Putative Cobalamin Riboswitch
In Vivo
Authors: Pérez Adam A. , Liu Zhenfeng , Rodionov Dmitry A. , Li Zhongkui , Bryant Donald A.
Published: July 2016

The euryhaline cyanobacterium
Synechococcus
sp. strain PCC 7002 has an obligate requirement for exogenous vitamin B
12
(cobalamin), but little is known about the roles of this compound in cyanobacteria. Bioinformatic analyses suggest that only the terminal enzyme in methionine biosynthesis, methionine synthase, requires cobalamin as a coenzyme in
Synechococcus
sp. strain PCC 7002. Methionine synthase (MetH) catalyzes the transfer of a methyl group from
N
5
-methyl-5,6,7,8-tetrahydrofolate to
l
-homocysteine during
l
-methionine synthesis and uses methylcobalamin as an intermediate methyl donor. Numerous bacteria and plants alternatively employ a cobalamin-independent methionine synthase isozyme, MetE, that catalyzes the same methyl transfer reaction as MetH but uses
N
5
-methyl-5,6,7,8-tetrahydrofolate directly as the methyl donor. The cobalamin auxotrophy of
Synechococcus
sp. strain PCC 7002 was complemented by using the
metE
gene from the closely related cyanobacterium
Synechococcus
sp. strain PCC 73109, which possesses genes for both methionine synthases. This result suggests that methionine biosynthesis is probably the sole use of cobalamin in
Synechococcus
sp. strain PCC 7002. Furthermore, a cobalamin-repressible gene expression system was developed in
Synechococcus
sp. strain PCC 7002 that was used to validate the presence of a cobalamin riboswitch in the promoter region of
metE
from
Synechococcus
sp. strain PCC 73109. This riboswitch acts as a cobalamin-dependent transcriptional attenuator for
metE
in that organism.

IMPORTANCE
Synechococcus
sp. strain PCC 7002 is a cobalamin auxotroph because, like eukaryotic marine algae, it uses a cobalamin-dependent methionine synthase (MetH) for the final step of
l
-methionine biosynthesis but cannot synthesize cobalamin
de novo
. Heterologous expression of
metE
, encoding cobalamin-independent methionine synthase, from
Synechococcus
sp. strain PCC 73109, relieved this auxotrophy and enabled the construction of a truly autotrophic
Synechococcus
sp. strain PCC 7002 more suitable for large-scale industrial applications. Characterization of a cobalamin riboswitch expands the genetic toolbox for
Synechococcus
sp. strain PCC 7002 by providing a cobalamin-repressible expression system.
Stair climbing/descending exercise for a short time decreases blood glucose levels after a meal in people with type 2 diabetes
Authors: Honda Hiroto , Igaki Makoto , Hatanaka Yuki , Komatsu Motoaki , Tanaka Shin-ichiro , Miki Tetsuo , Suzuki Taiga , Takaishi Tetsuo , Hayashi Tatsuya
Published: July 2016

Objective

We examined whether stair climbing–descending exercise (ST-EX), a convenient method to increase physical activity in daily life, for a short period would acutely improve the postprandial blood glucose (BG) response in people with type 2 diabetes (T2D).

Methods

16 people with T2D (age 65.4±1.1 years) participated in 2 separate sessions. After an overnight fast, each participant consumed a test meal and then kept resting for 180 min, except when performing each 3 min bout of ST-EX at 60 and 120 min after the meal (ST-EX session), or kept resting for 180 min (REST session). ST-EX comprised 6 continuous repetitions of climbing to the second floor (21 steps) at a rate of 80–110 steps/min followed by walking down slowly to the first floor at a free step rate.

Results

The BG at 60 min after the meal during the ST-EX session (immediately before the first ST-EX) did not differ from that during the REST session, but analysis of variance revealed a significant interaction between time and treatment (p<0.01). The BG at 150 min after the meal (30 min after the second ST-EX) was significantly lower than that during the REST session (p<0.01). The area under the curve was also 18% lower during the ST-EX session than during the REST session (p<0.05). The heart rate and blood lactate levels indicated that the actual intensity of ST-EX was ‘hard’. In contrast, the rating of perceived exertion (RPE) indicated that the overall intensity of ST-EX was ‘moderate’ because of decreased RPE scores during descent.

Conclusions

The present findings suggest that performing 3 min ST-EX 60 and 120 min after a meal may be a useful strategy to accelerate the decrease in postprandial BG levels in people with T2D.
Pilot study to test the feasibility of a trial design and complex intervention onPRIoritisingMUltimedication inMultimorbidity in general practices (PRIMUMpilot)
Authors: Muth Christiane , Harder Sebastian , Uhlmann Lorenz , Rochon Justine , Fullerton Birgit , Güthlin Corina , Erler Antje , Beyer Martin , van den Akker Marjan , Perera Rafael , Knottnerus André , Valderas Jose M , Gerlach Ferdinand M , Haefeli Walter E
Published: July 2016

Objective

To improve medication appropriateness and adherence in elderly patients with multimorbidity, we developed a complex intervention involving general practitioners (GPs) and their healthcare assistants (HCA). In accordance with the Medical Research Council guidance on developing and evaluating complex interventions, we prepared for the main study by testing the feasibility of the intervention and study design in a cluster randomised pilot study.

Setting

20 general practices in Hesse, Germany.

Participants

100 cognitively intact patients ≥65 years with ≥3 chronic conditions, ≥5 chronic prescriptions and capable of participating in telephone interviews; 94 patients completed the study.

Intervention

The HCA conducted a checklist-based interview with patients on medication-related problems and reconciled their medications. Assisted by a computerised decision-support system (CDSS), the GPs discussed medication intake with patients and adjusted their medication regimens. The control group continued with usual care.

Outcome measures

Feasibility of the intervention and required time were assessed for GPs, HCAs and patients using mixed methods (questionnaires, interviews and case vignettes after completion of the study). The feasibility of the study was assessed concerning success of achieving recruitment targets, balancing cluster sizes and minimising drop-out rates. Exploratory outcomes included the medication appropriateness index (MAI), quality of life, functional status and adherence-related measures. MAI was evaluated blinded to group assignment, and intra-rater/inter-rater reliability was assessed for a subsample of prescriptions.

Results

10 practices were randomised and analysed per group. GPs/HCAs were satisfied with the interventions despite the time required (35/45 min/patient). In case vignettes, GPs/HCAs needed help using the CDSS. The study made no patients feel uneasy. Intra-rater/inter-rater reliability for MAI was excellent. Inclusion criteria were challenging and potentially inadequate, and should therefore be adjusted. Outcome measures on pain, functionality and self-reported adherence were unfeasible due to frequent missing values, an incorrect manual or potentially invalid results.

Conclusions

Intervention and trial design were feasible. The pilot study revealed important limitations that influenced the design and conduct of the main study, thus highlighting the value of piloting complex interventions.

Trial registration number

ISRCTN99691973; Results.