bluuebunny/crossref_metadata_2025 huggingface dataset:
The
mtfA
Transcription Factor Gene Controls Morphogenesis, Gliotoxin Production, and Virulence in the Opportunistic Human Pathogen Aspergillus fumigatus
Authors: Smith Timothy D. , Calvo Ana M.
Published: April 2014

Aspergillus fumigatus
is the leading causative agent of invasive aspergillosis (IA). The number of cases is on the rise, with mortality rates as high as 90% among immunocompromised patients. Molecular genetic studies in
A. fumigatus
could provide novel targets to potentially set the basis for antifungal therapies. In the current study, we investigated the role of the transcription factor gene
mtfA
in
A. fumigatus
. Our results revealed that
mtfA
plays a role in the growth and development of the fungus. Deletion or overexpression of
mtfA
leads to a slight reduction in colony growth, as well as a reduction in conidiation levels, in the overexpression strain compared to the wild-type strain. Furthermore, production of the secondary metabolite gliotoxin increased when
mtfA
was overexpressed, coinciding with an increase in the transcription levels of the gliotoxin genes
gliZ
and
gliP
with respect to the wild type. In addition, our study showed that
mtfA
is also necessary for normal protease activity in
A. fumigatus
; deletion of
mtfA
resulted in a reduction of protease activity compared to wild-type levels. Importantly, the absence of
mtfA
caused a decrease in virulence in the
Galleria mellonella
infection model, indicating that
mtfA
is necessary for
A. fumigatus
wild-type pathogenesis.
Identification and Characterization of a Gene Cluster Required for Proper Rod Shape, Cell Division, and Pathogenesis in Clostridium difficile
Authors: Ransom Eric M. , Williams Kyle B. , Weiss David S. , Ellermeier Craig D.
Published: April 2014

Little is known about cell division in
Clostridium difficile
, a strict anaerobe that causes serious diarrheal diseases in people whose normal intestinal microbiome has been perturbed by treatment with broad-spectrum antibiotics. Here we identify and characterize a gene cluster encoding three cell division proteins found only in
C. difficile
and a small number of closely related bacteria. These proteins were named MldA, MldB, and MldC, for
m
idcell
l
ocalizing
d
ivision proteins. MldA is predicted to be a membrane protein with coiled-coil domains and a peptidoglycan-binding SPOR domain. MldB and MldC are predicted to be cytoplasmic proteins; MldB has two predicted coiled-coil domains, but MldC lacks obvious conserved domains or sequence motifs. Mutants of
mldA
or
mldB
had morphological defects, including loss of rod shape (a curved cell phenotype) and inefficient separation of daughter cells (a chaining phenotype). Fusions of cyan fluorescent protein (CFP) to MldA, MldB, and MldC revealed that all three proteins localize sharply to the division site. This application of CFP was possible because we discovered that O
2
-dependent fluorescent proteins produced anaerobically can acquire fluorescence after cells are fixed with cross-linkers to preserve native patterns of protein localization. Mutants lacking the Mld proteins are severely attenuated for pathogenesis in a hamster model of
C. difficile
infection. Because all three Mld proteins are essentially unique to
C. difficile
, they might be exploited as targets for antibiotics that combat
C. difficile
without disrupting the intestinal microbiome.
Construction and Characterization of
tetH
Overexpression and Knockout Strains of Acidithiobacillus ferrooxidans
Authors: Yu Yangyang , Liu Xiangmei , Wang Huiyan , Li Xiuting , Lin Jianqun
Published: April 2014

Acidithiobacillus ferrooxidans
is a major participant in consortia of microorganisms used for bioleaching. It can obtain energy from the oxidation of Fe
2+
, H
2
, S
0
, and various reduced inorganic sulfur compounds (RISCs). Tetrathionate is a key intermediate during RISC oxidation, hydrolyzed by tetrathionate hydrolase (TetH), and used as sole energy source. In this study, a
tetH
knockout (Δ
tetH
) mutant and a
tetH
overexpression strain were constructed and characterized. The
tetH
overexpression strain grew better on sulfur and tetrathionate and possessed a higher rate of tetrathionate utilization and TetH activity than the wild type. However, its cell yields on tetrathionate were much lower than those on sulfur. The Δ
tetH
mutant could not grow on tetrathionate but could proliferate on sulfur with a lower cell yield than the wild type's, which indicated that tetrathionate hydrolysis is mediated only by TetH, encoded by
tetH
. The Δ
tetH
mutant could survive in ferrous medium with an Fe
2+
oxidation rate similar to that of the wild type. For the
tetH
overexpression strain, the rate was relatively higher than that of the wild type. The reverse transcription-quantitative PCR (qRT-PCR) results showed that
tetH
and
doxD2
acted synergistically, and
doxD2
was considered important in thiosulfate metabolism. Of the two
sqr
genes, AFE_0267 seemed to play as important a role in sulfide oxidation as AFE_1792. This study not only provides a substantial basis for studying the function of the
tetH
gene but also may serve as a model to clarify other candidate genes involved in sulfur oxidation in this organism.
Adaptation of Iron Homeostasis Pathways by a Pseudomonas aeruginosa Pyoverdine Mutant in the Cystic Fibrosis Lung
Authors: Nguyen Angela T. , O'Neill Maura J. , Watts Annabelle M. , Robson Cynthia L. , Lamont Iain L. , Wilks Angela , Oglesby-Sherrouse Amanda G.
Published: April 2014

Cystic fibrosis (CF) patients suffer from chronic bacterial lung infections, most notably by
Pseudomonas aeruginosa
, which persists for decades in the lungs and undergoes extensive evolution.
P. aeruginosa
requires iron for virulence and uses the fluorescent siderophore pyoverdine to scavenge and solubilize ferric iron during acute infections. Pyoverdine mutants accumulate in the lungs of some CF patients, however, suggesting that the heme and ferrous iron acquisition pathways of
P. aeruginosa
are more important in this environment. Here, we sought to determine how evolution of
P. aeruginosa
in the CF lung affects iron acquisition and regulatory pathways through the use of longitudinal CF isolates. These analyses demonstrated a significant reduction of siderophore production during the course of CF lung infection in nearly all strains tested. Mass spectrometry analysis of one of these strains showed that the later CF isolate has streamlined the metabolic flux of extracellular heme through the HemO heme oxygenase, resulting in more-efficient heme utilization. Moreover, gene expression analysis shows that iron regulation via the PrrF small RNAs (sRNAs) is enhanced in the later CF isolate. Finally, analysis of
P. aeruginosa
gene expression in the lungs of various CF patients demonstrates that both PrrF and HemO are consistently expressed in the CF lung environment. Combined, these results suggest that heme is a critical source of iron during prolonged infection of the CF lung and that changes in iron and heme regulatory pathways play a crucial role in adaptation of
P. aeruginosa
to this ever-changing host environment.
CugP Is a Novel Ubiquitous Non-GalU-Type Bacterial UDP-Glucose Pyrophosphorylase Found in Cyanobacteria
Authors: Maeda Kaisei , Narikawa Rei , Ikeuchi Masahiko
Published: April 2014

UDP-glucose pyrophosphorylase synthesizes UDP-glucose from UTP and glucose 1-phosphate and exists in almost all species. Most bacteria possess a GalU-type UDP-glucose pyrophosphorylase, whereas many cyanobacteria species do not. In certain cyanobacteria, UDP-glucose is used as a substrate for synthesis of exopolysaccharide cellulose in spite of the absence of GalU-type UDP-glucose pyrophosphorylase. Therefore, there should be an uncharacterized UDP-glucose pyrophosphorylase in cyanobacteria. Here, we show that all cyanobacteria possess a non-GalU-type bacterial UDP-glucose pyrophosphorylase, i.e., CugP, a novel family in the nucleotide triphosphate transferase superfamily. The expressed recombinant
Synechocystis
sp. strain PCC 6803 CugP had pyrophosphorylase activity that was highly specific for UTP and glucose 1-phosphate. The fact that the CugP gene cannot be deleted completely in
Synechocystis
sp. PCC 6803 suggests its central role as the substrate supplier for galactolipid synthesis. Galactolipids are major constituents of the photosynthetic thylakoid membrane and important for photosynthetic activity. Based on phylogenetic analysis, this CugP-type UDP-glucose pyrophosphorylase may have recently been horizontally transferred to certain noncyanobacteria.
Amiodarone-induced myxoedema coma
Authors: Hassan Syed , Ayoub Walaa , Hassan Mona , Wisgerhof Max
Published: April 2014

A 62-year-old man was found to have bradycardia, hypothermia and respiratory failure 3 weeks after initiation of amiodarone therapy for atrial fibrillation. Thyroid-stimulating hormone was found to be 168 μIU/mL (nl. 0.3–5 μIU/mL) and free thyroxine (FT4) was <0.2 ng/dL (nl. 0.8–1.8 ng/dL). He received intravenous fluids, vasopressor therapy and stress dose steroids; he was intubated and admitted to the intensive care unit. He received 500 μg of intravenous levothyroxine in the first 18 h of therapy, and 150 µg intravenous daily thereafter. Haemodynamic improvement, along with complete recovery of mental status, occurred after 48 h. Twelve hours after the initiation of therapy, FT4 was 0.96 ng/dL. The patient was maintained on levothyroxine 175 (g POorally daily. A thyroid ultrasound showed diffuse heterogeneity. The 24 hour excretion of iodine was 3657 (mcg (25–756 ( mcg). The only two cases of amiodarone-induced myxoedema coma in the literature report patient death despite supportive therapy and thyroid hormone replacement. This case represents the most thoroughly investigated case of amiodarone-induced myxoedema coma with a history significant for subclinical thyroid disease.
Metastatic bone lesion due to methotrexate and etanercept 24 years after breast cancer treatment
Authors: Yano Shuichi
Published: April 2014

A 72-year-old woman with rheumatoid arthritis presented with lumbar vertebral bone metastasis 24 years after mammectomy and radiotherapy for breast cancer. She was treated with prednisolone and methotrexate (MTX) for 11 months to which 10 mg of etanercept twice a week was added for a further 8 months. On the basis of this result, the possibility of a metastatic bone lesion appearing many years after cancer treatment should be considered when planning MTX and etanercept therapy.
Acute double flexor tendon ruptures following injection of collagenase clostridium histolyticum (Xiapex) for Dupuytren's contracture
Authors: Povlsen Bo , Singh Sarvpreet
Published: April 2014

We report a case of acute (24 h) double flexor tendon rupture of the little finger after a single injection of collagenase clostridium histolyticum into a palmar Dupuytren's contracture cord which caused metacarpophalangeal joint contracture. Tendon surgery was performed 48 h postinjury with primary repair and standard rehabilitation but it resulted in poor active flexion due to adhesions. Previous papers have suggested that a needle inserted into the flexor tendon can be detected prior to the injection of collagenase by asking the patient to actively move the finger, but our test on an awake patient showed that when a 27-gauge needle was inserted into the flexor tendons through a thick palmar cord, the syringe did not move significantly when the patient moved the finger, and therefore this test does not minimise the risk of iatrogenic tendon injury when using collagenase (Xiapex) for Dupuytren's contracture.
Acute myocardial infarction following honeybee sting
Authors: Puttegowda Beeresh , Chikkabasavaiah Nagamani , Basavappa Ramesh , Akmal Khateeb Syed Tanveer
Published: April 2014

The occurrence of acute myocardial infarction following a honeybee sting has been very rarely reported in the previous literature. Possible pathogenetic mechanisms include severe hypotension, rarely hypertension and coronary vasospasm with subsequent thrombosis of coronary vessels developed after the release of vasoactive, inflammatory and thrombogenic substances contained in the bee venom. This syndrome is also known as Kounis syndrome. We report a case of a 32-year-old man who presented with acute inferior wall myocardial infarction within 3 h of honeybee sting. He was treated with thrombolytic therapy. Coronary angiogram revealed normal coronary arteries.
Amelanotic malignant melanoma of the cervical oesophagus
Authors: Ramaswamy Balakrishnan , Bhandarkar Ajay M , Venkitachalam Shruti , Trivedi Shivangi
Published: April 2014

We report the case of a young woman who presented with progressive dysphagia and swelling in the anterior aspect of the neck of short duration. On evaluation, she was diagnosed with amelanotic malignant melanoma of the cervical oesophagus. She underwent total laryngopharyngo-oesophagectomy with gastric transposition with bilateral modified radical neck dissection with feeding jejunostomy and a permanent tracheostomy with postoperative combined chemoradiation therapy. However, in spite of aggressive treatment, the patient expired 8 months after initial presentation with distant metastasis.
Understanding the role of embarrassment in gynaecological screening: a qualitative study from the ASPIRE cervical cancer screening project in Uganda
Authors: Teng Flora F , Mitchell Sheona M , Sekikubo Musa , Biryabarema Christine , Byamugisha Josaphat K , Steinberg Malcolm , Money Deborah M , Ogilvie Gina S
Published: April 2014

Objective

To define embarrassment and develop an understanding of the role of embarrassment in relation to cervical cancer screening and self-collected human papillomavirus (HPV) DNA testing in Uganda.

Design

Cross-sectional, qualitative study using semistructured one-to-one interviews and focus groups.

Participants

6 key-informant health workers and 16 local women, purposively sampled. Key informant inclusion criteria: Ugandan members of the project team. Focus group inclusion criteria: woman age 30–69 years, Luganda or Swahili speaking, living or working in the target Ugandan community. Exclusion criteria: unwillingness to sign informed consent.

Setting

Primary and tertiary low-resource setting in Kampala, Uganda.

Results

In Luganda, embarrassment relating to cervical cancer is described in two forms. ‘Community embarrassment’ describes discomfort based on how a person may be perceived by others. ‘Personal embarrassment’ relates to shyness or discomfort with her own genitalia. Community embarrassment was described in themes relating to place of study recruitment, amount of privacy in dwellings, personal relationship with health workers, handling of the vaginal swab and misunderstanding of HPV self-collection as HIV testing. Themes of personal embarrassment related to lack of knowledge, age and novelty of the self-collection swab. Overall, embarrassment was a barrier to screening at the outset and diminished over time through education and knowledge. Fatalism regarding cervical cancer diagnosis, worry about results and stigma associated with a cervical cancer diagnosis were other psychosocial barriers described. Overcoming psychosocial barriers to screening can include peer-to-peer education, drama and media campaigns.

Conclusions

Embarrassment and other psychosocial barriers may play a large role at the onset of a screening programme, but over time as education and knowledge increase, and the social norms around screening evolve, its role diminishes. The role of peer-to-peer education and community authorities on healthcare cannot be overlooked and can have a major impact in overcoming psychosocial and social barriers to screening.
Patient and public involvement in research and the Cancer Experiences Collaborative: benefits and challenges
Authors: Froggatt Katherine , Preston Nancy , Turner Mary , Kerr Chris
Published: April 2014

Aims and objectives

The involvement of patients in the design and conduct of research is increasingly promoted by policy and research bodies. The experiences of individuals who become involved in research in this way are not well understood. This study aims to describe the experiences of people's participation in patient and public involvement (PPI) in supportive and palliative care research, specifically with respect to the benefits and challenges of participation for the individuals and the broader research support structures.

Methods

In this qualitative exploratory study, semistructured interviews were undertaken with individuals who had a cancer diagnosis and were involved in a supportive and palliative care research collaborative over a period of 6 years. Recruitment was through the host university organisations involved in the research collaborative. A thematic analysis was undertaken to identify commonalities and differences across their experiences.

Findings

The eight participants in the study were highly motivated and undertook PPI in research alongside other voluntary activities. They identified a number of research and personal benefits: bringing a lay perspective into research, making a difference and personal gains. Personal gains concerned support, new knowledge and skills and greater confidence. The challenges to participation that were identified included the emotional and practical nature of the work, issues of language and identifying the difference made.

Conclusions

Involvement in supportive and palliative care research is valued by people with a cancer diagnosis, but there are ongoing issues of emotional support and good communication through the use of appropriate language in documentation.
On Block-transitive 6-(v, k, λ)-Designs with k at Most 10000
Authors: Tan Qionghua , Liu Weijun , Chen Jing
Published: April 2014

In recent years, there has been a great deal of research concerning the flag-transitive t-designs, however, few about the block-transitive t-designs with t large (t ≥ 4). In 1993, Cameron and Praeger conjectured that there are no non-trivial block-transitive 6-designs. In this paper, we prove that the conjecture is true when k ≤ 10000 and [Formula: see text] is almost simple.
Scaling laws for aggradation, denudation and progradation rates: the case for time-scale invariance at sediment sources and sinks
Authors: Sadler Peter M. , Jerolmack Douglas J.
Published: April 2014

Linear rates of sediment aggradation and fluvial incision are inverse functions of measurement interval, a generic consequence of unsteadiness in the underlying processes. This effect results from a one-dimensional approach–that is, vertical rates determined at a single location–and significantly complicates comparisons of rates at different timescales. Mass conservation imposes an important but underutilized constraint; sediment by-passing or eroded from one location must deposit somewhere else. Over the long term, sediment generation and deposition must balance. In principle, the effects of unsteadiness could be eliminated if the total volume of sediment eroded or deposited over different intervals could be measured. In practice, however, obtaining such three-dimensional data from an individual site is virtually impossible. Here, we advance from one- to two-dimensional rate data. We present two new global compilations of data: denudation rates of fluvial uplands; and lateral migration (progradation) rates of siliciclastic lowland and marine systems, from ripple to shelf-slope scale. Important new findings are: (1) upland denudation rates determined from specific sediment yield show little or no dependence of rate on time interval; (2) in the transfer zone between sediment source and sink, rates of erosion and deposition balance over all scales; and (3) progradation mirrors aggradation over all timescales. The product of progradation and aggradation is independent of timescale, implying that global sediment flux into the world’s oceans has been constant on the order of 10
0
m
2
/yr, from scales of months to tens of millions of years. Results show that global rates of denudation and accumulation are time invariant with appropriate spatial averaging; however, site-specific application remains a daunting challenge.
Chronic inhibition of phosphodiesterase 5 with tadalafil attenuates mitochondrial dysfunction in type 2 diabetic hearts: potential role of NO/SIRT1/PGC-1α signaling
Authors: Koka Saisudha , Aluri Hema S. , Xi Lei , Lesnefsky Edward J. , Kukreja Rakesh C.
Published: April 2014

Enhanced nitric oxide (NO) production is known to activate silent information regulator 1 (SIRT1), which is a histone deacetylase that regulates PGC-1α, a regulator of mitochondrial biogenesis and coactivator of transcription factors impacting energy homeostasis. Since phosphodiesterase-5 inhibitors potentiate NO signaling, we hypothesized that chronic treatment with phosphodiesterase-5 inhibitor tadalafil would activate SIRT1-PGC-1α signaling and protect against metabolic stress-induced mitochondrial dysfunction in diabetic hearts. Diabetic db/db mice ( n = 32/group; 40 wk old) were randomized to receive DMSO (10%, 0.2 ml ip) or tadalafil (1 mg/kg ip in 10% DMSO) for 8 wk. Wild-type C57BL mice served as nondiabetic controls. The hearts were excised and homogenized to study SIRT1 activity and downstream protein targets. Mitochondrial function was determined by measuring oxidative phosphorylation (OXPHOS), and reactive oxygen species generation was studied in isolated mitochondria. Tadalafil-treated diabetic mice demonstrated significantly improved left ventricular function, which is associated with increased cardiac SIRT1 activity. Tadalafil also enhanced plasma NO oxidation levels, myocardial SIRT1, PGC-1α expression, and phosphorylation of eNOS, Akt, and AMPK in the diabetic hearts. OXPHOS with the complex I substrate glutamate was decreased by 50% in diabetic hearts compared with the nondiabetic controls. Tadalafil protected OXPHOS with an improved glutamate state 3 respiration rates. The increased reactive oxygen species production from complex I was significantly decreased by tadalafil treatment. In conclusion, chronic treatment with tadalafil activates NO-induced SIRT1-PGC-1α signaling and attenuates mitochondrial dysfunction in type 2 diabetic hearts.
Pioglitazone reduces angiotensin II-induced COX-2 expression through inhibition of ROS production and ET-1 transcription in vascular cells from spontaneously hypertensive rats
Authors: Pérez-Girón Jose V. , Palacios Roberto , Martín Angela , Hernanz Raquel , Aguado Andrea , Martínez-Revelles Sonia , Barrús María T. , Salaices Mercedes , Alonso María J.
Published: April 2014

Glitazones have anti-inflammatory properties by interfering with the transcription of proinflammatory genes, such as cyclooxygenase (COX)-2, and with ROS production, which are increased in hypertension. This study analyzed whether pioglitazone modulates COX-2 expression in hypertension by interfering with ROS and endothelin (ET)-1. In vivo, pioglitazone (2.5 mg·kg−1·day−1, 28 days) reduced the greater levels of COX-2, pre-pro-ET-1, and NADPH oxidase (NOX) expression and activity as well as O2·− production found in aortas from spontaneously hypertensive rats (SHRs). ANG II increased COX-2 and pre-pro-ET-1 levels more in cultured vascular smooth muscle cells from hypertensive rats compared with normotensive rats. The ETA receptor antagonist BQ-123 reduced ANG II-induced COX-2 expression in SHR cells. ANG II also increased NOX-1 expression, NOX activity, and superoxide production in SHR cells; the selective NOX-1 inhibitor ML-171 and catalase reduced ANG II-induced COX-2 and ET-1 transcription. ANG II also increased c-Jun transcription and phospho-JNK1/2, phospho-c-Jun, and p65 NF-κB subunit nuclear protein expression. SP-600125 and lactacystin, JNK and NF-κB inhibitors, respectively, reduced ANG II-induced ET-1, COX-2, and NOX-1 levels and NOX activity. Pioglitazone reduced the effects of ANG II on NOX activity, NOX-1, pre-pro-ET-1, COX-2, and c-Jun mRNA levels, JNK activation, and nuclear phospho-c-Jun and p65 expression. In conclusion, ROS production and ET-1 are involved in ANG II-induced COX-2 expression in SHRs, explaining the greater COX-2 expression observed in this strain. Furthermore, pioglitazone inhibits ANG II-induced COX-2 expression likely by interfering with NF-κB and activator protein-1 proinflammatory pathways and downregulating ROS production and ET-1 transcription, thus contributing to the anti-inflammatory properties of glitazones.
Lung volume quantified by MRI reflects extracellular-matrix deposition and altered pulmonary function in bleomycin models of fibrosis: effects of SOM230
Authors: Egger Christine , Gérard Christelle , Vidotto Nella , Accart Nathalie , Cannet Catherine , Dunbar Andrew , Tigani Bruno , Piaia Alessandro , Jarai Gabor , Jarman Elizabeth , Schmid Herbert A. , Beckmann Nicolau
Published: April 2014

Idiopathic pulmonary fibrosis is a progressive and lethal disease, characterized by loss of lung elasticity and alveolar surface area, secondary to alveolar epithelial cell injury, reactive inflammation, proliferation of fibroblasts, and deposition of extracellular matrix. The effects of oropharyngeal aspiration of bleomycin in Sprague-Dawley rats and C57BL/6 mice, as well as of intratracheal administration of ovalbumin to actively sensitized Brown Norway rats on total lung volume as assessed noninvasively by magnetic resonance imaging (MRI) were investigated here. Lung injury and volume were quantified by using nongated or respiratory-gated MRI acquisitions [ultrashort echo time (UTE) or gradient-echo techniques]. Lung function of bleomycin-challenged rats was examined additionally using a flexiVent system. Postmortem analyses included histology of collagen and hydroxyproline assays. Bleomycin induced an increase of MRI-assessed total lung volume, lung dry and wet weights, and hydroxyproline content as well as collagen amount. In bleomycin-treated rats, gated MRI showed an increased volume of the lung in the inspiratory and expiratory phases of the respiratory cycle and a temporary decrease of tidal volume. Decreased dynamic lung compliance was found in bleomycin-challenged rats. Bleomycin-induced increase of MRI-detected lung volume was consistent with tissue deposition during fibrotic processes resulting in decreased lung elasticity, whereas influences by edema or emphysema could be excluded. In ovalbumin-challenged rats, total lung volume quantified by MRI remained unchanged. The somatostatin analog, SOM230, was shown to have therapeutic effects on established bleomycin-induced fibrosis in rats. This work suggests MRI-detected total lung volume as readout for tissue-deposition in small rodent bleomycin models of pulmonary fibrosis.
Accumulation of Memory Precursor CD8 T Cells in Regressing Tumors following Combination Therapy with Vaccine and Anti-PD-1 Antibody
Authors: Karyampudi Lavakumar , Lamichhane Purushottam , Scheid Adam D. , Kalli Kimberly R. , Shreeder Barath , Krempski James W. , Behrens Marshall D. , Knutson Keith L.
Published: April 2014

Immunosuppression in the tumor microenvironment blunts vaccine-induced immune effectors. PD-1/B7-H1 is an important inhibitory axis in the tumor microenvironment. Our goal in this study was to determine the effect of blocking this inhibitory axis during and following vaccination against breast cancer. We observed that using anti-PD-1 antibody and a multipeptide vaccine (consisting of immunogenic peptides derived from breast cancer antigens, neu, legumain, and β-catenin) as a combination therapy regimen for the treatment of breast cancer–bearing mice prolonged the vaccine-induced progression-free survival period. This prolonged survival was associated with increase in number of Tc1 and Tc2 CD8 T cells with memory precursor phenotype, CD27+IL-7RhiT-betlo, and decrease in number of PD-1+ dendritic cells (DC) in regressing tumors and enhanced antigen reactivity of tumor-infiltrating CD8 T cells. It was also observed that blockade of PD-1 on tumor DCs enhanced IL-7R expression on CD8 T cells. Taken together, our results suggest that PD-1 blockade enhances breast cancer vaccine efficacy by altering both CD8 T cell and DC components of the tumor microenvironment. Given the recent success of anti-PD-1 monotherapy, our results are encouraging for developing combination therapies for the treatment of patients with cancer in which anti-PD-1 monotherapy alone may be ineffective (i.e., PD-L1–negative tumors). Cancer Res; 74(11); 2974–85. ©2014 AACR.
P-Glycoprotein, CYP3A, and Plasma Carboxylesterase Determine Brain and Blood Disposition of the mTOR Inhibitor Everolimus (Afinitor) in Mice
Authors: Tang Seng Chuan , Sparidans Rolf W. , Cheung Ka Lei , Fukami Tatsuki , Durmus Selvi , Wagenaar Els , Yokoi Tsuyoshi , van Vlijmen Bart J.M. , Beijnen Jos H. , Schinkel Alfred H.
Published: April 2014

Purpose: To clarify the role of ABCB1, ABCG2, and CYP3A in blood and brain exposure of everolimus using knockout mouse models.

Experimental Design: We used wild-type, Abcb1a/1b−/−, Abcg2−/−, Abcb1a/1b;Abcg2−/−, and Cyp3a−/− mice to study everolimus oral bioavailability and brain accumulation.

Results: Following everolimus administration, brain concentrations and brain-to-liver ratios were substantially increased in Abcb1a/1b−/−and Abcb1a/1b;Abcg2−/−, but not Abcg2−/−mice. The fraction of everolimus located in the plasma compartment was highly increased in all knockout strains. In vitro, everolimus was rapidly degraded in wild-type but not knockout plasma. Carboxylesterase 1c (Ces1c), a plasma carboxylesterase gene, was highly upregulated (∼80-fold) in the liver of knockout mice relative to wild-type mice, and plasma Ces1c likely protected everolimus from degradation by binding and stabilizing it. This binding was prevented by preincubation with the carboxylesterase inhibitor BNPP. In vivo knockdown experiments confirmed the involvement of Ces1c in everolimus stabilization. Everolimus also markedly inhibited the hydrolysis of irinotecan and p-nitrophenyl acetate by mouse plasma carboxylesterase and recombinant human CES2, respectively. After correcting for carboxylesterase binding, Cyp3a−/−, but not Abcb1a/1b−/−, Abcg2−/−, or Abcb1a/1b;Abcg2−/−mice, displayed highly (>5-fold) increased oral availability of everolimus.

Conclusions: Brain accumulation of everolimus was restricted by Abcb1, but not Abcg2, suggesting the use of coadministered ABCB1 inhibitors to improve brain tumor treatment. Cyp3a, but not Abcb1a/1b, restricted everolimus oral availability, underscoring drug–drug interaction risks via CYP3A. Upregulated Ces1c likely mediated the tight binding and stabilization of everolimus, causing higher plasma retention in knockout strains. This Ces upregulation might confound other pharmacologic studies. Clin Cancer Res; 20(12); 3133–45. ©2014 AACR.
CADM1 Expression and Stepwise Downregulation of CD7 Are Closely Associated with Clonal Expansion of HTLV-I–Infected Cells in Adult T-cell Leukemia/Lymphoma
Authors: Kobayashi Seiichiro , Nakano Kazumi , Watanabe Eri , Ishigaki Tomohiro , Ohno Nobuhiro , Yuji Koichiro , Oyaizu Naoki , Asanuma Satomi , Yamagishi Makoto , Yamochi Tadanori , Watanabe Nobukazu , Tojo Arinobu , Watanabe Toshiki , Uchimaru Kaoru
Published: April 2014

Purpose: Cell adhesion molecule 1 (CADM1), initially identified as a tumor suppressor gene, has recently been reported to be ectopically expressed in primary adult T-cell leukemia–lymphoma (ATL) cells. We incorporated CADM1 into flow-cytometric analysis to reveal oncogenic mechanisms in human T-cell lymphotrophic virus type I (HTLV-I) infection by purifying cells from the intermediate stages of ATL development.

Experimental Design: We isolated CADM1- and CD7-expressing peripheral blood mononuclear cells of asymptomatic carriers and ATLs using multicolor flow cytometry. Fluorescence-activated cell sorted (FACS) subpopulations were subjected to clonal expansion and gene expression analysis.

Results: HTLV-I–infected cells were efficiently enriched in CADM1+ subpopulations (D, CADM1posCD7dim and N, CADM1posCD7neg). Clonally expanding cells were detected exclusively in these subpopulations in asymptomatic carriers with high proviral load, suggesting that the appearance of D and N could be a surrogate marker of progression from asymptomatic carrier to early ATL. Further disease progression was accompanied by an increase in N with a reciprocal decrease in D, indicating clonal evolution from D to N. The gene expression profiles of D and N in asymptomatic carriers showed similarities to those of indolent ATLs, suggesting that these subpopulations represent premalignant cells. This is further supported by the molecular hallmarks of ATL, that is, drastic downregulation of miR-31 and upregulation of abnormal Helios transcripts.

Conclusion: The CADM1 versus CD7 plot accurately reflects disease progression in HTLV-I infection, and CADM1+ cells with downregulated CD7 in asymptomatic carriers have common properties with those in indolent ATLs. Clin Cancer Res; 20(11); 2851–61. ©2014 AACR.
Druggable Oncogene Fusions in Invasive Mucinous Lung Adenocarcinoma
Authors: Nakaoku Takashi , Tsuta Koji , Ichikawa Hitoshi , Shiraishi Kouya , Sakamoto Hiromi , Enari Masato , Furuta Koh , Shimada Yoko , Ogiwara Hideaki , Watanabe Shun-ichi , Nokihara Hiroshi , Yasuda Kazuki , Hiramoto Masaki , Nammo Takao , Ishigame Teruhide , Schetter Aaron J. , Okayama Hirokazu , Harris Curtis C. , Kim Young Hak , Mishima Michiaki , Yokota Jun , Yoshida Teruhiko , Kohno Takashi
Published: April 2014

Purpose: To identify druggable oncogenic fusions in invasive mucinous adenocarcinoma (IMA) of the lung, a malignant type of lung adenocarcinoma in which KRAS mutations frequently occur.

Experimental Design: From an IMA cohort of 90 cases, consisting of 56 cases (62%) with KRAS mutations and 34 cases without (38%), we conducted whole-transcriptome sequencing of 32 IMAs, including 27 cases without KRAS mutations. We used the sequencing data to identify gene fusions, and then performed functional analyses of the fusion gene products.

Results: We identified oncogenic fusions that occurred mutually exclusively with KRAS mutations: CD74-NRG1, SLC3A2-NRG1, EZR-ERBB4, TRIM24-BRAF, and KIAA1468-RET. NRG1 fusions were present in 17.6% (6/34) of KRAS-negative IMAs. The CD74-NRG1 fusion activated HER2:HER3 signaling, whereas the EZR-ERBB4 and TRIM24-BRAF fusions constitutively activated the ERBB4 and BRAF kinases, respectively. Signaling pathway activation and fusion-induced anchorage-independent growth/tumorigenicity of NIH3T3 cells expressing these fusions were suppressed by tyrosine kinase inhibitors approved for clinical use.

Conclusions: Oncogenic fusions act as driver mutations in IMAs without KRAS mutations, and thus represent promising therapeutic targets for the treatment of such IMAs. Clin Cancer Res; 20(12); 3087–93. ©2014 AACR.
TLR7 Ligand Augments GM-CSF–Initiated Antitumor Immunity through Activation of Plasmacytoid Dendritic Cells
Authors: Narusawa Megumi , Inoue Hiroyuki , Sakamoto Chika , Matsumura Yumiko , Takahashi Atsushi , Inoue Tomoko , Watanabe Ayumi , Miyamoto Shohei , Miura Yoshie , Hijikata Yasuki , Tanaka Yoshihiro , Inoue Makoto , Takayama Koichi , Okazaki Toshihiko , Hasegawa Mamoru , Nakanishi Yoichi , Tani Kenzaburo
Published: April 2014

Vaccination with irradiated granulocyte macrophage colony-stimulating factor (GM-CSF)–transduced autologous tumor cells (GVAX) has been shown to induce therapeutic antitumor immunity. However, its effectiveness is limited. We therefore attempted to improve the antitumor effect by identifying little-known key pathways in GM-CSF–sensitized dendritic cells (GM-DC) in tumor-draining lymph nodes (TDLN). We initially confirmed that syngeneic mice subcutaneously injected with poorly immunogenic Lewis lung carcinoma (LLC) cells transduced with Sendai virus encoding GM-CSF (LLC/SeV/GM) remarkably rejected the tumor growth. Using cDNA microarrays, we found that expression levels of type I interferon (IFN)–related genes, predominantly expressed in plasmacytoid DCs (pDC), were significantly upregulated in TDLN-derived GM-DCs and focused on pDCs. Indeed, mouse experiments demonstrated that the effective induction of GM-CSF–induced antitumor immunity observed in immunocompetent mice treated with LLC/SeV/GM cells was significantly attenuated when pDC-depleted or IFNα receptor knockout (IFNAR−/−) mice were used. Importantly, in both LLC and CT26 colon cancer–bearing mice, the combinational use of imiquimod with autologous GVAX therapy overcame the refractoriness to GVAX monotherapy accompanied by tolerability. Mechanistically, mice treated with the combined vaccination displayed increased expression levels of CD86, CD9, and Siglec-H, which correlate with an antitumor phenotype, in pDCs, but decreased the ratio of CD4+CD25+FoxP3+ regulatory T cells in TDLNs. Collectively, these findings indicate that the additional use of imiquimod to activate pDCs with type I IFN production, as a positive regulator of T-cell priming, could enhance the immunologic antitumor effects of GVAX therapy, shedding promising light on the understanding and treatment of GM-CSF–based cancer immunotherapy. Cancer Immunol Res; 2(6); 568–80. ©2014 AACR.
Fli1 Acts Downstream of Etv2 to Govern Cell Survival and Vascular Homeostasis via Positive Autoregulation
Authors: Abedin Md J. , Nguyen Annie , Jiang Nan , Perry Cameron E. , Shelton John M. , Watson Dennis K. , Ferdous Anwarul
Published: April 2014

Rationale:

Cardiovascular health depends on proper development and integrity of blood vessels. Ets variant 2 (Etv2), a member of the E26 transforming–specific family of transcription factors, is essential to initiate a transcriptional program leading to vascular morphogenesis in early mouse embryos. However, endothelial expression of the
Etv2
gene ceases at midgestation; therefore, vascular development past this stage must continue independent of Etv2.

Objective:

To identify molecular mechanisms underlying transcriptional regulation of vascular morphogenesis and homeostasis in the absence of Etv2.

Methods and Results:

Using loss- and gain-of-function strategies and a series of molecular techniques, we identify Friend leukemia integration 1 (
Fli1
), another E26 transforming–specific family transcription factor, as a downstream target of Etv2. We demonstrate that Etv2 binds to conserved Ets-binding sites within the promoter region of the
Fli1
gene and governs
Fli1
expression. Importantly, in the absence of Etv2 at midgestation, binding of Etv2 at Ets-binding sites in the
Fli1
promoter is replaced by Fli1 protein itself, sustaining expression of
Fli1
as well as selective Etv2-regulated endothelial genes to promote endothelial cell survival and vascular integrity. Consistent with this, we report that Fli1 binds to the conserved Ets-binding sites within promoter and enhancer regions of other Etv2-regulated endothelial genes, including
Tie2
, to control their expression at and beyond midgestation.

Conclusions:

We have identified a novel positive feed-forward regulatory loop in which Etv2 activates expression of genes involved in vasculogenesis, including
Fli1
. Once the program is activated in early embryos, Fli1 then takes over to sustain the process in the absence of Etv2.
Early Results of Massachusetts Healthcare Reform on Racial, Ethnic, and Socioeconomic Disparities in Cardiovascular Care
Authors: Albert Michelle A. , Ayanian John Z. , Silbaugh Treacy S. , Lovett Ann , Resnic Fred , Jacobs Aryana , Normand Sharon-Lise T.
Published: April 2014

Background—

Insured adults receive invasive cardiovascular procedures more frequently than uninsured adults. We examined the impact of healthcare reform in Massachusetts on use of coronary revascularization procedures and in-hospital and 1-year mortality by race/ethnicity, education, and sex.

Methods and Results—

Using hospital claims data, we compared differences in coronary revascularization rates (coronary artery bypass grafting or percutaneous coronary intervention) and in-hospital mortality by race/ethnicity, education, and sex among Massachusetts residents aged 21 to 64 years hospitalized with a principal discharge diagnosis of ischemic heart disease before (November 1, 2004, to July 31, 2006) and after (December 1, 2006, to September 30, 2008) reform; 1-year mortality was calculated for those undergoing revascularization. Adjusted logistic regression assessed 24 216 discharges before reform and 20 721 discharges after reform. Blacks had 30% lower odds of receiving coronary revascularization than whites in the prereform period. Compared with whites in the postreform period, blacks (odds ratio=0.73; 95% confidence interval, 0.63–0.84) and Hispanics (odds ratio= 0.84; 95% confidence interval, 0.74–0.97) were less likely and Asians (odds ratio=1.29; 95% confidence interval, 1.01–1.65) were more likely to receive coronary revascularization. Patients living in more educated communities, men, and persons with private insurance were more likely to receive coronary revascularization before and after reform. Compared with the prereform period, the adjusted odds of in-hospital mortality were higher in patients living in less-educated communities in the postreform period. No differences in 1-year mortality by race/ethnicity, education, or sex for revascularized patients were observed before or after reform.

Conclusions—

Reducing insurance barriers to receipt of coronary revascularization procedures has not yet eliminated preexisting demographic and educational disparities in access to these procedures.
The spatial concentration of bias: An examination of the community factors that influence residents’ perceptions of bias crime
Authors: Sydes Michelle L , Wickes Rebecca L , Higginson Angela
Published: April 2014

Emerging scholarship indicates that bias crimes are concentrated in particular types of places. Currently, only a small number of studies consider the ecological factors that influence official reports of bias crime. Results from these studies indicate that the community processes and structures associated with the occurrence of non-bias crime may operate differently for bias crime. We use administrative and survey data from approximately 4000 residents living across 148 communities in Brisbane, Queensland to examine the ecological drivers of bias crime. Using multi-level logistic regression, we examine the community and household factors associated with residents’ perceptions of bias crime. Here, we focus not only on the structural demographics of the community, but also on the degree to which community cohesion influences whether or not residents perceive bias crime as a problem in their community. We find that poverty and ethnic diversity are positively associated with residents’ perceptions of bias crime. Further, residents living in communities with higher levels of community cohesion are less likely to perceive bias crime as a problem in their community. The level of community cohesion fully mediates the impact of ethnic diversity and partially mediates the effect of poverty on residents’ perceptions of bias crime.
“The Discipline Stop”
Authors: Brockenbrough Ed
Published: April 2014

Calls for the recruitment and retention of more Black male teachers have unfolded amid popular depictions of Black men as patriarchal disciplinarians. Against that backdrop, this article investigates how 11 Black male teachers were positioned as disciplinary agents in a predominantly Black urban school district on the east coast of the United States. As they described the discipline-related expectations they encountered in their jobs, study participants critiqued and complicated prevailing perceptions of Black male teachers as authoritarian disciplinarians for Black students. Through careful analyses of participants’ narratives, this article offers new insights into how Black male teachers negotiate their roles as disciplinarians, and it raises several questions that could drive future efforts to understand and support Black male teachers’ disciplinary practices in today’s urban schools.
The invisible flâneuse: European cities and the African sex worker in Chika Unigwe’s On Black Sisters’ Street
Authors: Bastida-Rodriguez Patricia
Published: April 2014

Recent migration to Europe has become the focus of some interesting fiction produced in the last decade. One such narrative is On Black Sisters’ Street (2009) by Nigerian-born Belgian writer Chika Unigwe, which deals with the experiences of undocumented African immigrants in the Flemish city of Antwerp who must survive as sex workers in order to pay the human trafficking networks that brought them to Europe. However, the novel does not only stand as a testimony of a sad social reality but also as an exploration of urban space, urban movement, and subjectivity in contemporary European cities. This article examines the intersections between the protagonists’ use of urban space, their social status as prostitutes, and the emotions circulating about them in the city, since they will lead to relevant insights about contemporary urban movement and its literary representation.
Effects of nanoclay and coupling agent on fungal degradation and water absorption of sanding dust/high density polyethylene composites
Authors: Ashori Alireza , Shahrebabak Amineh Behzadi , Madhoushi Mehrab
Published: April 2014

In this study, the effects of two variable parameters namely nanoclay and coupling agent on the durability against fungal attack and water absorption of wood plastic composites (WPCs) were investigated. Composites based on high density polyethylene (HDPE), sanding dusts (SDs), and nanoclay (montmorillonite type) were made by melt compounding and then hot-pressing. Maleic anhydride grafted polyethylene (MAPE) was used as coupling agent. Treated and untreated (control) composites were exposed to the fungal decay using white-rot fungus ( Trametes versicolor) for 4, 8, and 12 weeks according to the modified ASTM D 1413 standard. The experimental results indicated that the exposure of the composites to a five-cycle of boiling and drying caused serious damage to the interfacial adhesion between SDs and HDPE matrix due to contraction and swelling stresses developed during the cyclic immersion. The composites filled with nanoclay were more resistant than the control sample to decay. However, based on statistical analysis significant differences in water absorption of the composites were not observed after addition of nanoclay. In general, fungal degradation and water absorption were significantly decreased by the incorporation of MAPE. However, addition of 6 wt% MAPE improved both properties more than 3 wt% MAPE. Morphological studies showed fungal colonization and some micro voids, which accelerate water absorption. In conclusion, this work has shown that addition of nanoclay and coupling agent can considerably decrease fungal decay and water absorption of WPCs.
Contact and Compromise
Authors: Pickett Justin T. , Baker Thomas , Metcalfe Christi , Gertz Marc , Bellandi Rose
Published: April 2014

Objectives:

Informed by intergroup contact theory, this study explores the relationships between intergroup contact, perceived out-group threat, and support for conciliatory solutions to the violent conflict between Israeli Jews and Palestinians in Israel and the Palestinian territories.

Methods:

Regression and structural equation models analyze public opinion data collected in Israel in 2011 and 2012. The analyses assess whether quantity and quality of Israeli Jews’ contact with Israeli Arabs in day-to-day encounters are associated with their support for conciliatory policies.

Results:

The quality, but not the quantity, of contact is associated with lower levels of perceived Palestinian threat and, in turn, with increased support for compromise.

Conclusion:

The current study provides initial evidence that everyday interactions with Israeli Arabs, when they occur under optimal conditions, may have the potential to reduce Israeli Jews’ perceptions of Palestinian threat and, in turn, increase their support for compromise.