bluuebunny/crossref_metadata_2025 huggingface dataset:
Block copolymer self-assembly–directed synthesis of mesoporous gyroidal superconductors
Authors: Robbins Spencer W. , Beaucage Peter A. , Sai Hiroaki , Tan Kwan Wee , Werner Jörg G. , Sethna James P. , DiSalvo Francis J. , Gruner Sol M. , Van Dover Robert B. , Wiesner Ulrich
Published: January 2016

Block copolymer self-assembly is used to synthesize three-dimensionally continuous gyroidal mesoporous superconductors of niobium nitride.
HOX
s and lincRNAs: Two sides of the same coin
Authors: De Kumar Bony , Krumlauf Robb
Published: January 2016

The Hox complexes: Domains of dynamic interplay between coding and noncoding transcripts.
Airborne and Grain Dust Fungal Community Compositions Are Shaped Regionally by Plant Genotypes and Farming Practices
Authors: Pellissier Loïc , Oppliger Anne , Hirzel Alexandre H. , Savova-Bianchi Dessislava , Mbayo Guilain , Mascher Fabio , Kellenberger Stefan , Niculita-Hirzel Hélène
Published: January 2016

Chronic exposure to airborne fungi has been associated with different respiratory symptoms and pathologies in occupational populations, such as grain workers. However, the homogeneity in the fungal species composition of these bioaerosols on a large geographical scale and the different drivers that shape these fungal communities remain unclear. In this study, the diversity of fungi in grain dust and in the aerosols released during harvesting was determined across 96 sites at a geographical scale of 560 km
2
along an elevation gradient of 500 m by tag-encoded 454 pyrosequencing of the internal transcribed spacer (ITS) sequences. Associations between the structure of fungal communities in the grain dust and different abiotic (farming system, soil characteristics, and geographic and climatic parameters) and biotic (wheat cultivar and previous crop culture) factors were explored. These analyses revealed a strong relationship between the airborne and grain dust fungal communities and showed the presence of allergenic and mycotoxigenic species in most samples, which highlights the potential contribution of these fungal species to work-related respiratory symptoms of grain workers. The farming system was the major driver of the alpha and beta phylogenetic diversity values of fungal communities. In addition, elevation and soil CaCO
3
concentrations shaped the alpha diversity, whereas wheat cultivar, cropping history, and the number of freezing days per year shaped the taxonomic beta diversity of these communities.
Activity-Based Protein Profiling of Ammonia Monooxygenase in Nitrosomonas europaea
Authors: Bennett Kristen , Sadler Natalie C. , Wright Aaron T. , Yeager Chris , Hyman Michael R.
Published: January 2016

Nitrosomonas europaea
is an aerobic nitrifying bacterium that oxidizes ammonia (NH
3
) to nitrite (NO
2
−
) through the sequential activities of ammonia monooxygenase (AMO) and hydroxylamine dehydrogenase (HAO). Many alkynes are mechanism-based inactivators of AMO, and here we describe an activity-based protein profiling method for this enzyme using 1,7-octadiyne (17OD) as a probe. Inactivation of NH
4
+
-dependent O
2
uptake by
N. europaea
by 17OD was time- and concentration-dependent. The effects of 17OD were specific for ammonia-oxidizing activity, and
de novo
protein synthesis was required to reestablish this activity after cells were exposed to 17OD. Cells were reacted with Alexa Fluor 647 azide using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) (click) reaction, solubilized, and analyzed by SDS-PAGE and infrared (IR) scanning. A fluorescent 28-kDa polypeptide was observed for cells previously exposed to 17OD but not for cells treated with either allylthiourea or acetylene prior to exposure to 17OD or for cells not previously exposed to 17OD. The fluorescent polypeptide was membrane associated and aggregated when heated with β-mercaptoethanol and SDS. The fluorescent polypeptide was also detected in cells pretreated with other diynes, but not in cells pretreated with structural homologs containing a single ethynyl functional group. The membrane fraction from 17OD-treated cells was conjugated with biotin-azide and solubilized in SDS. Streptavidin affinity-purified polypeptides were on-bead trypsin-digested, and amino acid sequences of the peptide fragments were determined by liquid chromatography-mass spectrometry (LC-MS) analysis. Peptide fragments from AmoA were the predominant peptides detected in 17OD-treated samples. In-gel digestion and matrix-assisted laser desorption ionization–tandem time of flight (MALDI-TOF/TOF) analyses also confirmed that the fluorescent 28-kDa polypeptide was AmoA.
Lack of Involvement of Fenton Chemistry in Death of Methicillin-Resistant and Methicillin-Sensitive Strains of Staphylococcus aureus and Destruction of Their Genomes on Wet or Dry Copper Alloy Surfaces
Authors: Warnes Sarah L. , Keevil C. William
Published: January 2016

The pandemic of hospital-acquired infections caused by methicillin-resistant
Staphylococcus aureus
(MRSA) has declined, but the evolution of strains with enhanced virulence and toxins and the increase of community-associated infections are still a threat. In previous studies, 10
7
MRSA bacteria applied as simulated droplet contamination were killed on copper and brass surfaces within 90 min. However, contamination of surfaces is often via finger tips and dries rapidly, and it may be overlooked by cleaning regimes (unlike visible droplets). In this new study, a 5-log reduction of a hardy epidemic strain of MRSA (epidemic methicillin-resistant
S. aureus
16 [EMRSA-16]) was observed following 10 min of contact with copper, and a 4-log reduction was observed on copper nickel and cartridge brass alloys in 15 min. A methicillin-sensitive
S. aureus
(MSSA) strain from an osteomyelitis patient was killed on copper surfaces in 15 min, and 4-log and 3-log reductions occurred within 20 min of contact with copper nickel and cartridge brass, respectively. Bacterial respiration was compromised on copper surfaces, and superoxide was generated as part of the killing mechanism. In addition, destruction of genomic DNA occurs on copper and brass surfaces, allaying concerns about horizontal gene transfer and copper resistance. Incorporation of copper alloy biocidal surfaces may help to reduce the spread of this dangerous pathogen.
Skills retention in Sudanese village midwives 1 year following Helping Babies Breathe training
Authors: ME Arabi Ali , Ibrahim Salah A , Ahmed Sami E , MacGinnea Finn , Hawkes Gavin , Dempsey Eugene , Anthony Ryan C
Published: January 2016

Background

Over 80% of deliveries in Sudan occur in isolated villages, attended by village midwives (VMWs). Upgrading newborn resuscitation skills with the Helping Babies Breathe (HBB) programme could improve newborn survival rates.

Objective

To describe the competencies in newborn resuscitation of selected VMWs pre-HBB and post-HBB training.

Methods

In a prospective intervention study, the VMWs’ performances in the HBB Objective Structured Clinical Examination B simulated scenario (manikin requiring face-mask ventilation (FMV)) were digitally recorded and analysed prior to and 3 and 12 months following HBB training. Regular manikin-based practice was encouraged following training.

Results

Pre-HBB training, 42% of 71 VMWs (of whom 61% were functionally illiterate) stimulated the non-breathing manikin by holding it by the legs and either stimulated/slapped (30.4%) or shook (12.7%) it, while 25% (18/71) provided manikin mouth-to-mouth ventilation. The low scorings on the ‘preparation for birth’ (0% and 3.1% at 3 and 12 months, respectively) were mainly due to failure to demonstrate the subitem of ‘cleans hands’. The percentage of VMWs providing manikin FMV within the Golden Minute increased from 37.3% (25/67) to 72.3% (47/65) (p<0.005), but there were no significant differences in the number of VMWs producing at least five FMVs at 3 months (73%, 49/67) and 12 months (58%, 38/65), respectively.

Conclusions

VMWs, despite a high illiteracy rate, absorbed and sustained HBB skills for at least a year. Regular, low intensity, manikin-based skills training with peers may have helped sustain FMV, but not hand-cleansing skills.
Trends in market share of leading cigarette brands in the USA: national survey on drug use and health 2002–2013
Authors: Sharma Anushree , Fix Brian V , Delnevo Cristine , Cummings K Michael , O'Connor Richard J
Published: January 2016

Objectives

The main objective of this study is to examine trends in market share for leading cigarette brands, both before (2002–2008) and after (2009–2013) Food and Drug Administration (FDA) regulation of tobacco products.

Design Data come from the annual National Survey on Drug Use and Health from 2002 through 2013. Descriptive statistics, cross tabulations, and logistic regression were employed. Data were weighted to the US population and adjusted for cigarette consumption. Our analysis is restricted to 164 343 current cigarette smokers who were at least 12 years of age or older, had smoked at least one cigarette in the 30 days prior to the survey, and reported a usual cigarette brand at the time of the survey.

Results

Over 12 years, 14 brands comprised over 77% of the cigarette market. Marlboro consistently held over 38% of the market. Newport held the second highest market share, and increased from 7.2% in 2002 to 10.9% by 2013. Market share of Pall Mall grew by over 400% (1.7% in 2002 vs 8.9% in 2013), likely aided by the 2009 Federal excise tax increase. No clear associations of changes in market share with the implementation of FDA's regulatory authority over tobacco in 2009 were noted.

Conclusions

Tracking market share trends offers clues about brand marketing changing preferences of consumers. Rapidly growing cigarette brands should be monitored to determine if specific marketing practices or design changes are drivers, as these could represent public health concerns. Monitoring trends in cigarette market share could inform regulatory decision-making efforts related to marketing and advertising.
Association of restaurant smoking ban and the incidence of acute myocardial infarction in Finland
Authors: Sipilä Jussi Olli Tapani , Gunn Jarmo Mikael , Kauko Tommi , Rautava Päivi , Kytö Ville
Published: January 2016

Objective

To describe the changes in nationwide acute myocardial infarction (AMI) incidence following the implementation of a law banning smoking indoors in restaurants on 1 June 2007.

Methods

Retrospective registry study of all hospitalisations for AMI in Finland. All 34 887 hospitalisations for AMI between 1 June 2005 and 31 May 2009 were identified from the Care Register for Health Care (CRHC) and statistics for tobacco consumption were obtained from the National Institute for Health and Welfare. Comorbidities for individual hospitalisations were searched from the CRHC.

Results

The incidence rate of AMI was reduced by 6.3% (95% CI 4.1% to 8.6%; p<0.0001) in the latter half of the study period following the smoking ban when adjusted for age, gender and overall population prevalence of smoking. Short-term incidence of AMI (6-month prior vs 6 months after the smoking ban) was also reduced (4.5%, 95% CI 0.2% to 9.0%; p=0.0399) and was largest in the working middle-aged group (40–50 years) but observed also in the oldest age group (>70 years). The incidence rates declined similarly for men and women.

Conclusions

Banning indoor tobacco smoking in restaurants was associated with a mild additional reduction in AMI incidence on a nationwide level in Finland.
Qualitative evaluation of the Safety and Improvement in Primary Care (SIPC) pilot collaborative in Scotland: perceptions and experiences of participating care teams
Authors: Bowie Paul , Halley Lyn , Blamey Avril , Gillies Jill , Houston Neil
Published: January 2016

Objectives

To explore general practitioner (GP) team perceptions and experiences of participating in a large-scale safety and improvement pilot programme to develop and test a range of interventions that were largely new to this setting.

Design

Qualitative study using semistructured interviews. Data were analysed thematically.

Subjects and setting

Purposive sample of multiprofessional study participants from 11 GP teams based in 3 Scottish National Health Service (NHS) Boards.

Results

27 participants were interviewed. 3 themes were generated: (1) programme experiences and benefits, for example, a majority of participants referred to gaining new theoretical and experiential safety knowledge (such as how unreliable evidence-based care can be) and skills (such as how to search electronic records for undetected risks) related to the programme interventions; (2) improvements to patient care systems, for example, improvements in care systems reliability using care bundles were reported by many, but this was an evolving process strongly dependent on closer working arrangements between clinical and administrative staff; (3) the utility of the programme improvement interventions, for example, mixed views and experiences of participating in the safety climate survey and meeting to reflect on the feedback report provided were apparent. Initial theories on the utilisation and potential impact of some interventions were refined based on evidence.

Conclusions

The pilot was positively received with many practices reporting improvements in safety systems, team working and communications with colleagues and patients. Barriers and facilitators were identified related to how interventions were used as the programme evolved, while other challenges around spreading implementation beyond this pilot were highlighted.
How do doctors and nurses manage delirium in intensive care units? A qualitative study using focus groups
Authors: Palacios-Ceña Domingo , Cachón-Pérez José Miguel , Martínez-Piedrola Rosa , Gueita-Rodriguez Javier , Perez-de-Heredia Marta , Fernández-de-las-Peñas Cesar
Published: January 2016

Objectives

The aim of this study was to explore the experiences of doctors and nurses caring for patients with delirium in the intensive care unit (ICU) and to describe the process of delirium management.

Setting

This study was performed in 5 ICUs located within 4 hospitals in Madrid (Spain).

Participants

Purposeful sampling was performed which included (1) doctors and nurses working in ICUs, (2) with >1 year experience in the ICU and (3) clinical experience with delirium. 38 professionals participated (19 doctors, 19 nurses), including 22 women and 16 men. The total mean age was 39 years.

Design

A qualitative study using focus groups.

Methods

7 focus groups were held to collect data: 3 nurse focus groups, 3 doctor focus groups and 1 mixed focus group. Each group comprised 6–10 participants. A semistructured questions guide was used. Thematic analysis methods were used to analyse the data.

Results

3 themes were identified: (1) the professional perspective on delirium; (2) implementing pharmacological and non-pharmacological treatment for delirium and (3) work organisation in the ICU. The professionals regarded patients with delirium with uncertainty, and felt they were often underdiagnosed and poorly managed. Doctors displayed discrepancies regarding pharmacological prescriptions and decision-making. The choice of medication was determined by experience. Nurses felt that, for many doctors, delirium was not considered a matter of urgency in the ICU. Nurses encountered difficulties when applying verbal restraint, managing sleep disorders and providing early mobilisation. The lack of a delirium protocol generates conflicts regarding what type of care management to apply, especially during the night shift. A degree of group pressure exists which, in turn, influences the decision-making process and patient care.

Conclusions

Patients with delirium represent complex cases, requiring the implementation of specific protocols. These results serve to improve the process of care in patients with delirium.
Improving the governance of patient safety in emergency care: a systematic review of interventions
Authors: Hesselink Gijs , Berben Sivera , Beune Thimpe , Schoonhoven Lisette
Published: January 2016

Objectives

To systematically review interventions that aim to improve the governance of patient safety within emergency care on effectiveness, reliability, validity and feasibility.

Design

A systematic review of the literature.

Methods

PubMed, EMBASE, Cumulative Index to Nursing and Allied Health Literature, the Cochrane Database of Systematic Reviews and PsychInfo were searched for studies published between January 1990 and July 2014. We included studies evaluating interventions relevant for higher management to oversee and manage patient safety, in prehospital emergency medical service (EMS) organisations and hospital-based emergency departments (EDs). Two reviewers independently selected candidate studies, extracted data and assessed study quality. Studies were categorised according to study quality, setting, sample, intervention characteristics and findings.

Results

Of the 18 included studies, 13 (72%) were non-experimental. Nine studies (50%) reported data on the reliability and/or validity of the intervention. Eight studies (44%) reported on the feasibility of the intervention. Only 4 studies (22%) reported statistically significant effects. The use of a simulation-based training programme and well-designed incident reporting systems led to a statistically significant improvement of safety knowledge and attitudes by ED staff and an increase of incident reports within EDs, respectively.

Conclusions

Characteristics of the interventions included in this review (eg, anonymous incident reporting and validation of incident reports by an independent party) could provide useful input for the design of an effective tool to govern patient safety in EMS organisations and EDs. However, executives cannot rely on a robust set of evidence-based and feasible tools to govern patient safety within their emergency care organisation and in the chain of emergency care. Established strategies from other high-risk sectors need to be evaluated in emergency care settings, using an experimental design with valid outcome measures to strengthen the evidence base.
Disease severity of Shiga toxin-producingE. coliO157 and factors influencing the development of typical haemolytic uraemic syndrome: a retrospective cohort study, 2009–2012
Authors: Launders N , Byrne L , Jenkins C , Harker K , Charlett A , Adak G K
Published: January 2016

Objectives

Assess the disease severity of Shiga toxin-producingEscherichia coli(STEC) O157 infection and factors influencing the development of typical haemolytic uraemic syndrome (tHUS).

Design

A retrospective cohort study using data collected through routine surveillance questionnaires between 2009 and 2012.

Participants

3323 symptomatic cases of STEC O157.

Main outcome measures

Incidence of human STEC O157 and tHUS, proportion of cases reporting bloody diarrhoea, hospitalisation, tHUS and death. Odds of progression to tHUS and predicted percentage chance of developing tHUS based on case demographics, STEC O157 strain characteristics and clinical symptoms.

Results

From 2009 to 2012, 3323 cases of symptomatic STEC O157 with completed questionnaires were reported, of which 172 developed tHUS (5.18%). Being aged 1–4 years (OR 8.65, 95% CI 5.01 to 14.94, p=0.004) or female (OR 1.61, 95% CI 1.12 to 2.30, p=0.009), being infected with phage type (PT) 21/28 (OR 2.07, 95% CI 1.25 to 3.42, p=0.005) or PT 2 (OR 2.18, 95% CI 1.06 to 4.50, p=0.034), receiving β-lactam antibiotics (OR 4.08, 95% CI 1.43 to 11.68, p=0.009) and presenting with vomiting (OR 3.16, 95% CI 2.16 to 4.62, p<0.001) or bloody diarrhoea (OR 2.10, 95% CI 1.38 to 3.20, p=0.001) were found to be significant risk factors for progression to tHUS. The predicted percentage chance of developing tHUS varied from under 1% to 50% if all risk factors were present.

Conclusions

The data from this study indicate the use of β-lactam antibiotics should be avoided in suspected cases of STEC infection in all age groups, particularly in those under the age of 5.
Is bias in the eye of the beholder? A vignette study to assess recognition of cognitive biases in clinical case workups
Authors: Zwaan Laura , Monteiro Sandra , Sherbino Jonathan , Ilgen Jonathan , Howey Betty , Norman Geoffrey
Published: January 2016

Background

Many authors have implicated cognitive biases as a primary cause of diagnostic error. If this is so, then physicians already familiar with common cognitive biases should consistently identify biases present in a clinical workup. The aim of this paper is to determine whether physicians agree on the presence or absence of particular biases in a clinical case workup and how case outcome knowledge affects bias identification.

Methods

We conducted a web survey of 37 physicians. Each participant read eight cases and listed which biases were present from a list provided. In half the cases the outcome implied a correct diagnosis; in the other half, it implied an incorrect diagnosis. We compared the number of biases identified when the outcome implied a correct or incorrect primary diagnosis. Additionally, the agreement among participants about presence or absence of specific biases was assessed.

Results

When the case outcome implied a correct diagnosis, an average of 1.75 cognitive biases were reported; when incorrect, 3.45 biases (F=71.3, p<0.00001). Individual biases were reported from 73% to 125% more often when an incorrect diagnosis was implied. There was no agreement on presence or absence of individual biases, with κ ranging from 0.000 to 0.044.

Interpretation

Individual physicians are unable to agree on the presence or absence of individual cognitive biases. Their judgements are heavily influenced by hindsight bias; when the outcome implies a diagnostic error, twice as many biases are identified. The results present challenges for current error reduction strategies based on identification of cognitive biases.
The short-term cost of falls, poisonings and scalds occurring at home in children under 5 years old in England: multicentre longitudinal study
Authors: Cooper N J , Kendrick D , Timblin C , Hayes M , Majsak-Newman G , Meteyard K , Hawkins A , Kay B
Published: January 2016

Background

Childhood falls, poisonings and scalds, occurring predominantly in the home, are an important public health problem, yet there is limited evidence on the costs of these injuries to individuals and society.

Objectives

To estimate National Health Service (NHS) and child and family costs of falls, poisonings and scalds.

Methods

We undertook a multicentre longitudinal study of falls, poisonings and scalds in children under 5 years old, set in acute NHS Trusts across four UK study centres. Data from parental self-reported questionnaires on health service resource use, family costs and expenditure were combined with unit cost data from published sources to calculate average cost for participants and injury mechanism.

Results

344 parents completed resource use questionnaires until their child recovered from their injury or until 12 months, whichever came soonest. Most injuries were minor, with >95% recovering within 2 weeks, and 99% within 1 month of the injury. 61% emergency department (ED) attendees were not admitted, 35% admitted for ≤1 day and 4% admitted for ≥2 days. The typical healthcare cost of an admission for ≥2 days was estimated at £2000–3000, for an admission for ≤1 day was £700–1000 and for an ED attendance without admission was £100–180. Family costs were considerable and varied across injury mechanisms. Of all injuries, scalds accrued highest healthcare and family costs.

Conclusions

Falls, poisonings and scalds incur considerable short-term healthcare and family costs. These data can inform injury prevention policy and commissioning of preventive services.
A length-based approach to predator–prey relationships in marine predators
Authors: Juanes Francis
Published: January 2016

Body size is a critical feature of the ecology of most organisms and has been used to describe and understand predator–prey interactions in both terrestrial and aquatic environments. Most previous studies have used prey mass to examine the relationships between predator size and prey size; however, using prey lengths may provide a different perspective, particularly for gape-limited fishes. Using a large database of predator and prey lengths for marine aquatic predators, I found the expected positive wedge-shaped relationship between predator length and prey length and a negative converging relationship between relative prey length (prey–predator length ratio = a measure of trophic niche breadth) and predator length. Distinct patterns in the size scaling of this measure of trophic niche breadth were identified using quantile regression: converging relationships were common among adults but absent among larvae. This difference suggests contrasting ontogenetic foraging opportunities between adults and larvae: a lack of large relative prey sizes for the largest adult predators, and a greater ability of larvae to include larger prey items in their diet as they grow.
Molecular mechanisms of STIM/Orai communication
Authors: Derler Isabella , Jardin Isaac , Romanin Christoph
Published: January 2016

Ca2+ entry into the cell via store-operated Ca2+ release-activated Ca2+ (CRAC) channels triggers diverse signaling cascades that affect cellular processes like cell growth, gene regulation, secretion, and cell death. These store-operated Ca2+ channels open after depletion of intracellular Ca2+ stores, and their main features are fully reconstituted by the two molecular key players: the stromal interaction molecule (STIM) and Orai. STIM represents an endoplasmic reticulum-located Ca2+ sensor, while Orai forms a highly Ca2+-selective ion channel in the plasma membrane. Functional as well as mutagenesis studies together with structural insights about STIM and Orai proteins provide a molecular picture of the interplay of these two key players in the CRAC signaling cascade. This review focuses on the main experimental advances in the understanding of the STIM1-Orai choreography, thereby establishing a portrait of key mechanistic steps in the CRAC channel signaling cascade. The focus is on the activation of the STIM proteins, the subsequent coupling of STIM1 to Orai1, and the consequent structural rearrangements that gate the Orai channels into the open state to allow Ca2+ permeation into the cell.
Endocannabinoids in cerebrovascular regulation
Authors: Benyó Zoltán , Ruisanchez Éva , Leszl-Ishiguro Miriam , Sándor Péter , Pacher Pál
Published: January 2016

The cerebral blood flow is tightly regulated by myogenic, endothelial, metabolic, and neural mechanisms under physiological conditions, and a large body of recent evidence indicates that inflammatory pathways have a major influence on the cerebral blood perfusion in certain central nervous system disorders, like hemorrhagic and ischemic stroke, traumatic brain injury, and vascular dementia. All major cell types involved in cerebrovascular control pathways (i.e., smooth muscle, endothelium, neurons, astrocytes, pericytes, microglia, and leukocytes) are capable of synthesizing endocannabinoids and/or express some or several of their target proteins [i.e., the cannabinoid 1 and 2 (CB1 and CB2) receptors and the transient receptor potential vanilloid type 1 ion channel]. Therefore, the endocannabinoid system may importantly modulate the regulation of cerebral circulation under physiological and pathophysiological conditions in a very complex manner. Experimental data accumulated since the late 1990s indicate that the direct effect of cannabinoids on cerebral vessels is vasodilation mediated, at least in part, by CB1 receptors. Cannabinoid-induced cerebrovascular relaxation involves both a direct inhibition of smooth muscle contractility and a release of vasodilator mediator(s) from the endothelium. However, under stress conditions (e.g., in conscious restrained animals or during hypoxia and hypercapnia), cannabinoid receptor activation was shown to induce a reduction of the cerebral blood flow, probably via inhibition of the electrical and/or metabolic activity of neurons. Finally, in certain cerebrovascular pathologies (e.g., subarachnoid hemorrhage, as well as traumatic and ischemic brain injury), activation of CB2 (and probably yet unidentified non-CB1/non-CB2) receptors appear to improve the blood perfusion of the brain via attenuating vascular inflammation.
RASSF1A Directly Antagonizes RhoA Activity through the Assembly of a Smurf1-Mediated Destruction Complex to Suppress Tumorigenesis
Authors: Lee Min-Goo , Jeong Seong-In , Ko Kyung-Phil , Park Soon-Ki , Ryu Byung-Kyu , Kim Ick-Young , Kim Jeong-Kook , Chi Sung-Gil
Published: January 2016

RASSF1A is a tumor suppressor implicated in many tumorigenic processes; however, the basis for its tumor suppressor functions are not fully understood. Here we show that RASSF1A is a novel antagonist of protumorigenic RhoA activity. Direct interaction between the C-terminal amino acids (256–277) of RASSF1A and active GTP-RhoA was critical for this antagonism. In addition, interaction between the N-terminal amino acids (69-82) of RASSF1A and the ubiquitin E3 ligase Smad ubiquitination regulatory factor 1 (Smurf1) disrupted GTPase activity by facilitating Smurf1-mediated ubiquitination of GTP-RhoA. We noted that the RhoA-binding domain of RASSF1A displayed high sequence homology with Rho-binding motifs in other RhoA effectors, such as Rhotekin. As predicted on this basis, RASSF1A competed with Rhotekin to bind RhoA and to block its activation. RASSF1A mutants unable to bind RhoA or Smurf1 failed to suppress RhoA-induced tumor cell proliferation, drug resistance, epithelial–mesenchymal transition, migration, invasion, and metastasis. Clinically, expression levels of RASSF1A and RhoA were inversely correlated in many types of primary and metastatic tumors and tumor cell lines. Collectively, our findings showed how RASSF1A may suppress tumorigenesis by intrinsically inhibiting the tumor-promoting activity of RhoA, thereby illuminating the potential mechanistic consequences of RASSF1A inactivation in many cancers. Cancer Res; 76(7); 1847–59. ©2016 AACR.
EGF816 Exerts Anticancer Effects in Non–Small Cell Lung Cancer by Irreversibly and Selectively Targeting Primary and Acquired Activating Mutations in the EGF Receptor
Authors: Jia Yong , Juarez Jose , Li Jie , Manuia Mari , Niederst Matthew J. , Tompkins Celin , Timple Noelito , Vaillancourt Mei-Ting , Pferdekamper AnneMarie Culazzo , Lockerman Elizabeth L. , Li Chun , Anderson Jennifer , Costa Carlotta , Liao Debbie , Murphy Eric , DiDonato Michael , Bursulaya Badry , Lelais Gerald , Barretina Jordi , McNeill Matthew , Epple Robert , Marsilje Thomas H. , Pathan Nuzhat , Engelman Jeffrey A. , Michellys Pierre-Yves , McNamara Peter , Harris Jennifer , Bender Steven , Kasibhatla Shailaja
Published: January 2016

Non–small cell lung cancer patients carrying oncogenic EGFR mutations initially respond to EGFR-targeted therapy, but later elicit minimal response due to dose-limiting toxicities and acquired resistance. EGF816 is a novel, irreversible mutant-selective EGFR inhibitor that specifically targets EGFR-activating mutations arising de novo and upon resistance acquisition, while sparing wild-type (WT) EGFR. EGF816 potently inhibited the most common EGFR mutations L858R, Ex19del, and T790M in vitro, which translated into strong tumor regressions in vivo in several patient-derived xenograft models. Notably, EGF816 also demonstrated antitumor activity in an exon 20 insertion mutant model. At levels above efficacious doses, EGF816 treatment led to minimal inhibition of WT EGFR and was well tolerated. In single-dose studies, EGF816 provided sustained inhibition of EGFR phosphorylation, consistent with its ability for irreversible binding. Furthermore, combined treatment with EGF816 and INC280, a cMET inhibitor, resulted in durable antitumor efficacy in a xenograft model that initially developed resistance to first-generation EGFR inhibitors via cMET activation. Thus, we report the first preclinical characterization of EGF816 and provide the groundwork for its current evaluation in phase I/II clinical trials in patients harboring EGFR mutations, including T790M. Cancer Res; 76(6); 1591–602. ©2016 AACR.
Impact of Treatment and Insurance on Socioeconomic Disparities in Survival after Adolescent and Young Adult Hodgkin Lymphoma: A Population-Based Study
Authors: Keegan Theresa H.M. , DeRouen Mindy C. , Parsons Helen M. , Clarke Christina A. , Goldberg Debbie , Flowers Christopher R. , Glaser Sally L.
Published: January 2016

Background: Previous studies documented racial/ethnic and socioeconomic disparities in survival after Hodgkin lymphoma among adolescents and young adults (AYA), but did not consider the influence of combined-modality treatment and health insurance.

Methods: Data for 9,353 AYA patients ages 15 to 39 years when diagnosed with Hodgkin lymphoma during 1988 to 2011 were obtained from the California Cancer Registry. Using multivariate Cox proportional hazards regression, we examined the impact of sociodemographic characteristics [race/ethnicity, neighborhood socioeconomic status (SES), and health insurance], initial combined-modality treatment, and subsequent cancers on survival.

Results: Over the 24-year study period, we observed improvements in Hodgkin lymphoma–specific survival by diagnostic period and differences in survival by race/ethnicity, neighborhood SES, and health insurance for a subset of more recently diagnosed patients (2001–2011). In multivariable analyses, Hodgkin lymphoma–specific survival was worse for Blacks than Whites with early-stage [HR: 1.68; 95% confidence interval (CI): 1.14–2.49] and late-stage disease (HR: 1.68; 95% CI, 1.17–2.41) and for Hispanics than Whites with late-stage disease (HR: 1.58; 95% CI, 1.22–2.04). AYAs diagnosed with early-stage disease experienced worse survival if they also resided in lower SES neighborhoods (HR: 2.06; 95% CI, 1.59–2.68). Furthermore, more recently diagnosed AYAs with public health insurance or who were uninsured experienced worse Hodgkin lymphoma–specific survival (HR: 2.08; 95% CI, 1.52–2.84).

Conclusion: Our findings identify several subgroups of Hodgkin lymphoma patients at higher risk for Hodgkin lymphoma mortality.

Impact: Identifying and reducing barriers to recommended treatment and surveillance in these AYAs at much higher risk of mortality is essential to ameliorating these survival disparities. Cancer Epidemiol Biomarkers Prev; 25(2); 264–73. ©2016 AACR.
Functional Analyses of Mutations in Receptor Tyrosine Kinase Genes in Non–Small Cell Lung Cancer: Double-Edged Sword of DDR2
Authors: Terashima Masato , Togashi Yosuke , Sato Katsuaki , Mizuuchi Hiroshi , Sakai Kazuko , Suda Kenichi , Nakamura Yu , Banno Eri , Hayashi Hidetoshi , De Velasco Marco A. , Fujita Yoshihiko , Tomida Shuta , Mitsudomi Tetsuya , Nishio Kazuto
Published: January 2016

Purpose: This study investigated whether mutations of receptor tyrosine kinase (RTK) genes detected using next-generation sequencing (NGS) are suitable therapeutic targets.

Experimental design: Fifty surgically resected non–small cell lung cancer (NSCLC) samples were target resequenced using NGS. We then investigated the functions of the identified RTK gene mutations, including their oncogenic potential, in vitro.

Results: Mutations in RTK genes were found in 20 samples (EGFR, 15; ERBB4, 1; ALK, 1; DDR2, 2; FGFR1, 1), mutations in MAPK pathway genes were found in nine samples (KRAS, 7; NRAS, 1; BRAF, 2), and mutations in PI3K pathway genes were found in three samples (PIK3CA, 1; PTEN, 3). Among the mutations in RTKs, the functions of four mutations were unclear (ERBB4 D245G; DDR2 H246R and E655K; FGFR1 A263V). These mutations did not exhibit any transformational activities. Neither the phosphorylation nor the protein expressions of RTKs were changed by the DDR2 H246R, ERBB4 D245G, and FGFR1 A263V mutations, although the expression level of the DDR2 protein harboring the E655K mutation was particularly low. Collagen stimulation decreased cellular proliferation through p38 activation in the DDR2 wild-type–overexpressed cell lines, whereas the growth-suppressive effect was weakened in DDR2 E655K–overexpressed cell lines. Furthermore, the DDR2 E655K protein strongly bound to ubiquitin ligase E3 (Cbl-b), and the mutant protein expression was increased after treatment with a proteasome inhibitor.

Conclusions: Our experimental findings suggest that RTK mutations are not always suitable as therapeutic targets. The DDR2 E655K mutation can play a role in cancer progression by reducing the growth-inhibitory effect of collagen. Clin Cancer Res; 22(14); 3663–71. ©2016 AACR.
Sequential Combination Therapy of CDK Inhibition and Doxorubicin Is Synthetically Lethal in p53-Mutant Triple-Negative Breast Cancer
Authors: Jabbour-Leung Natalie A. , Chen Xian , Bui Tuyen , Jiang Yufeng , Yang Dong , Vijayaraghavan Smruthi , McArthur Mark J. , Hunt Kelly K. , Keyomarsi Khandan
Published: January 2016

Triple-negative breast cancer (TNBC) is an aggressive malignancy in which the tumors lack expression of estrogen receptor, progesterone receptor, and HER2. Hence, TNBC patients cannot benefit from clinically available targeted therapies and rely on chemotherapy and surgery for treatment. While initially responding to chemotherapy, TNBC patients are at increased risk of developing distant metastasis and have decreased overall survival compared with non-TNBC patients. A majority of TNBC tumors carry p53 mutations, enabling them to bypass the G1 checkpoint and complete the cell cycle even in the presence of DNA damage. Therefore, we hypothesized that TNBC cells are sensitive to cell-cycle–targeted combination therapy, which leaves nontransformed cells unharmed. Our findings demonstrate that sequential administration of the pan-CDK inhibitor roscovitine before doxorubicin treatment is synthetically lethal explicitly in TNBC cells. Roscovitine treatment arrests TNBC cells in the G2–M cell-cycle phase, priming them for DNA damage. Combination treatment increased frequency of DNA double-strand breaks, while simultaneously reducing recruitment of homologous recombination proteins compared with doxorubicin treatment alone. Furthermore, this combination therapy significantly reduced tumor volume and increased overall survival compared with single drug or concomitant treatment in xenograft studies. Examination of isogenic immortalized human mammary epithelial cells and isogenic tumor cell lines found that abolishment of the p53 pathway is required for combination-induced cytotoxicity, making p53 a putative predictor of response to therapy. By exploiting the specific biologic and molecular characteristics of TNBC tumors, this innovative therapy can greatly impact the treatment and care of TNBC patients. Mol Cancer Ther; 15(4); 593–607. ©2016 AACR.
Gastric Adenocarcinomas Express the Glycosphingolipid Gb3/CD77: Targeting of Gastric Cancer Cells with Shiga Toxin B-Subunit
Authors: Geyer Philipp Emanuel , Maak Matthias , Nitsche Ulrich , Perl Markus , Novotny Alexander , Slotta-Huspenina Julia , Dransart Estelle , Holtorf Anne , Johannes Ludger , Janssen Klaus-Peter
Published: January 2016

The B-subunit of the bacterial Shiga toxin (STxB), which is nontoxic and has low immunogenicity, can be used for tumor targeting of breast, colon, and pancreatic cancer. Here, we tested whether human gastric cancers, which are among the most aggressive tumor entities, express the cellular receptor of Shiga toxin, the glycosphingolipid globotriaosylceramide (Gb3/CD77). The majority of cases showed an extensive staining for Gb3 (36/50 cases, 72%), as evidenced on tissue sections of surgically resected specimen. Gb3 expression was detected independent of type (diffuse/intestinal), and was negatively correlated to increasing tumor–node–metastasis stages (P = 0.0385), as well as with markers for senescence. Gb3 expression in nondiseased gastric mucosa was restricted to chief and parietal cells at the bottom of the gastric glands, and was not elevated in endoscopic samples of gastritis (n = 10). Gb3 expression in established cell lines of gastric carcinoma was heterogeneous, with 6 of 10 lines being positive, evidenced by flow cytometry. STxB was taken up rapidly by live Gb3-positive gastric cancer cells, following the intracellular retrograde transport route, avoiding lysosomes and rapidly reaching the Golgi apparatus and the endoplasmic reticulum. Treatment of the Gb3-expressing gastric carcinoma cell line St3051 with STxB coupled to SN38, the active metabolite of the topoisomerase type I inhibitor irinotecan, resulted in >100-fold increased cytotoxicity, as compared with irinotecan alone. No cytotoxicity was observed on gastric cancer cell lines lacking Gb3 expression, demonstrating receptor specificity of the STxB–SN38 compound. Thus, STxB is a highly specific transport vehicle for cytotoxic agents in gastric carcinoma. Mol Cancer Ther; 15(5); 1008–17. ©2016 AACR.
A Novel Recombinant Anti-CD22 Immunokinase Delivers Proapoptotic Activity of Death-Associated Protein Kinase (DAPK) and Mediates Cytotoxicity in Neoplastic B Cells
Authors: Lilienthal Nils , Lohmann Gregor , Crispatzu Giuliano , Vasyutina Elena , Zittrich Stefan , Mayer Petra , Herling Carmen Diana , Tur Mehmet Kemal , Hallek Michael , Pfitzer Gabriele , Barth Stefan , Herling Marco
Published: January 2016

The serine/threonine death-associated protein kinases (DAPK) provide pro-death signals in response to (oncogenic) cellular stresses. Lost DAPK expression due to (epi)genetic silencing is found in a broad spectrum of cancers. Within B-cell lymphomas, deficiency of the prototypic family member DAPK1 represents a predisposing or early tumorigenic lesion and high-frequency promoter methylation marks more aggressive diseases. On the basis of protein studies and meta-analyzed gene expression profiling data, we show here that within the low-level context of B-lymphocytic DAPK, particularly CLL cells have lost DAPK1 expression. To target this potential vulnerability, we conceptualized B-cell–specific cytotoxic reconstitution of the DAPK1 tumor suppressor in the format of an immunokinase. After rounds of selections for its most potent cytolytic moiety and optimal ligand part, a DK1KD-SGIII fusion protein containing a constitutive DAPK1 mutant, DK1KD, linked to the scFv SGIII against the B-cell–exclusive endocytic glyco-receptor CD22 was created. Its high purity and large-scale recombinant production provided a stable, selectively binding, and efficiently internalizing construct with preserved robust catalytic activity. DK1KD-SGIII specifically and efficiently killed CD22-positive cells of lymphoma lines and primary CLL samples, sparing healthy donor– or CLL patient–derived non-B cells. The mode of cell death was predominantly PARP-mediated and caspase-dependent conventional apoptosis as well as triggering of an autophagic program. The notoriously high apoptotic threshold of CLL could be overcome by DK1KD-SGIII in vitro also in cases with poor prognostic features, such as therapy resistance. The manufacturing feasibility of the novel CD22-targeting DAPK immunokinase and its selective antileukemic efficiency encourage intensified studies towards specific clinical application. Mol Cancer Ther; 15(5); 971–84. ©2016 AACR.
Preclinical Characterization of G1T28: A Novel CDK4/6 Inhibitor for Reduction of Chemotherapy-Induced Myelosuppression
Authors: Bisi John E. , Sorrentino Jessica A. , Roberts Patrick J. , Tavares Francis X. , Strum Jay C.
Published: January 2016

Chemotherapy-induced myelosuppression continues to represent the major dose-limiting toxicity of cytotoxic chemotherapy, which can be manifested as neutropenia, lymphopenia, anemia, and thrombocytopenia. As such, myelosuppression is the source of many of the adverse side effects of cancer treatment including infection, sepsis, bleeding, and fatigue, thus resulting in the need for hospitalizations, hematopoietic growth factor support, and transfusions (red blood cells and/or platelets). Moreover, clinical concerns raised by myelosuppression commonly lead to chemotherapy dose reductions, therefore limiting therapeutic dose intensity, and reducing the antitumor effectiveness of the treatment. Currently, the only course of treatment for myelosuppression is growth factor support which is suboptimal. These treatments are lineage specific, do not protect the bone marrow from the chemotherapy-inducing cytotoxic effects, and the safety and toxicity of each agent is extremely specific. Here, we describe the preclinical development of G1T28, a novel potent and selective CDK4/6 inhibitor that transiently and reversibly regulates the proliferation of murine and canine bone marrow hematopoietic stem and progenitor cells and provides multilineage protection from the hematologic toxicity of chemotherapy. Furthermore, G1T28 does not decrease the efficacy of cytotoxic chemotherapy on RB1-deficient tumors. G1T28 is currently in clinical development for the reduction of chemotherapy-induced myelosuppression in first- and second-line treatment of small-cell lung cancer. Mol Cancer Ther; 15(5); 783–93. ©2016 AACR.
EC-70124, a Novel Glycosylated Indolocarbazole Multikinase Inhibitor, Reverts Tumorigenic and Stem Cell Properties in Prostate Cancer by Inhibiting STAT3 and NF-κB
Authors: Civenni Gianluca , Longoni Nicole , Costales Paula , Dallavalle Cecilia , García Inclán Cristina , Albino Domenico , Nuñez Luz Elena , Morís Francisco , Carbone Giuseppina M. , Catapano Carlo V.
Published: January 2016

Cancer stem cells (CSC) contribute to disease progression and treatment failure in prostate cancer because of their intrinsic resistance to current therapies. The transcription factors NF-κB and STAT3 are frequently activated in advanced prostate cancer and sustain expansion of prostate CSCs. EC-70124 is a novel chimeric indolocarbazole compound generated by metabolic engineering of the biosynthetic pathways of glycosylated indolocarbazoles, such as staurosporine and rebeccamycin. In vitro kinome analyses revealed that EC-70124 acted as a multikinase inhibitor with potent activity against IKKβ and JAK2. In this study, we show that EC-70124 blocked concomitantly NF-κB and STAT3 in prostate cancer cells and particularly prostate CSCs, which exhibited overactivation of these transcription factors. Phosphorylation of IkB and STAT3 (Tyr705), the immediate targets of IKKβ and JAK2, respectively, was rapidly inhibited in vitro by EC-70124 at concentrations that were well below plasma levels in mice. Furthermore, the drug blocked activation of NF-κB and STAT3 reporters and suppressed transcription of their target genes. Treatment with EC-70124 impaired proliferation and colony formation in vitro and delayed development of prostate tumor xenografts. Notably, EC-70124 had profound effects on the prostate CSC subpopulation both in vitro and in vivo. Thus, EC-70124 is a potent inhibitor of the NF-κB and STAT3 signaling pathways and blocked tumor growth and maintenance of prostate CSCs. EC-70124 may provide the basis for developing new therapeutic strategies that combine agents directed to the CSC component and the bulk tumor cell population for treatment of advanced prostate cancer. Mol Cancer Ther; 15(5); 806–18. ©2016 AACR.