bluuebunny/crossref_metadata_2025 huggingface dataset:
Heat acclimation attenuates physiological strain and the HSP72, but not HSP90α, mRNA response to acute normobaric hypoxia
Authors: Gibson Oliver R. , Turner Gareth , Tuttle James A. , Taylor Lee , Watt Peter W. , Maxwell Neil S.
Published: July 2015

Heat acclimation (HA) attenuates physiological strain in hot conditions via phenotypic and cellular adaptation. The aim of this study was to determine whether HA reduced physiological strain, and heat shock protein (HSP) 72 and HSP90α mRNA responses in acute normobaric hypoxia. Sixteen male participants completed ten 90-min sessions of isothermic HA (40°C/40% relative humidity) or exercise training [control (CON); 20°C/40% relative humidity]. HA or CON were preceded (HYP1) and proceeded (HYP2) by a 30-min normobaric hypoxic exposure [inspired O2fraction = 0.12; 10-min rest, 10-min cycling at 40% peak O2uptake (V̇o2 peak), 10-min cycling at 65% V̇o2 peak]. HA induced greater rectal temperatures, sweat rate, and heart rates (HR) than CON during the training sessions. HA, but not CON, reduced resting rectal temperatures and resting HR and increased sweat rate and plasma volume. Hemoglobin mass did not change following HA nor CON. HSP72 and HSP90α mRNA increased in response to each HA session, but did not change with CON. HR during HYP2 was lower and O2saturation higher at 65% V̇o2 peakfollowing HA, but not CON. O2uptake/HR was greater at rest and 65% V̇o2 peakin HYP2 following HA, but was unchanged after CON. At rest, the respiratory exchange ratio was reduced during HYP2 following HA, but not CON. The increase in HSP72 mRNA during HYP1 did not occur in HYP2 following HA. In CON, HSP72 mRNA expression was unchanged during HYP1 and HYP2. In HA and CON, increases in HSP90α mRNA during HYP1 were maintained in HYP2. HA reduces physiological strain, and the transcription of HSP72, but not HSP90α mRNA in acute normobaric hypoxia.
The role of myostatin and activin receptor IIB in the regulation of unloading-induced myofiber type-specific skeletal muscle atrophy
Authors: Babcock Lyle W. , Knoblauch Mark , Clarke Mark S. F.
Published: July 2015

Chronic unloading induces decrements in muscle size and strength. This adaptation is governed by a number of molecular factors including myostatin, a potent negative regulator of muscle mass. Myostatin must first be secreted into the circulation and then bind to the membrane-bound activin receptor IIB (actRIIB) to exert its atrophic action. Therefore, we hypothesized that myofiber type-specific atrophy observed after hindlimb suspension (HLS) would be related to myofiber type-specific expression of myostatin and/or actRIIB. Wistar rats underwent HLS for 10 days, after which the tibialis anterior was harvested for frozen cross sectioning. Simultaneous multichannel immunofluorescent staining combined with differential interference contrast imaging was employed to analyze myofiber type-specific expression of myostatin and actRIIB and myofiber type cross-sectional area (CSA) across fiber types, myonuclei, and satellite cells. Hindlimb suspension (HLS) induced significant myofiber type-specific atrophy in myosin heavy chain (MHC) IIx ( P < 0.05) and MHC IIb myofibers ( P < 0.05). Myostatin staining associated with myonuclei was less in HLS rats compared with controls, while satellite cell staining for myostatin remained unchanged. In contrast, the total number myonuclei and satellite cells per myofiber was reduced in HLS compared with ambulatory control rats ( P < 0.01). Sarcoplasmic actRIIB staining differed between myofiber types (I < IIa < IIx < IIb) independent of loading conditions. Myofiber types exhibiting the greatest cytoplasmic staining of actRIIB corresponded to those exhibiting the greatest degree of atrophy following HLS. Our data suggest that differential expression of actRIIB may be responsible for myostatin-induced myofiber type-selective atrophy observed during chronic unloading.
Multineuronal activity patterns identify selective synaptic connections under realistic experimental constraints
Authors: Chambers Brendan , MacLean Jason N.
Published: July 2015

Structured multineuronal activity patterns within local neocortical circuitry are strongly linked to sensory input, motor output, and behavioral choice. These reliable patterns of pairwise lagged firing are the consequence of connectivity since they are not present in rate-matched but unconnected Poisson nulls. It is important to relate multineuronal patterns to their synaptic underpinnings, but it is unclear how effectively statistical dependencies in spiking between neurons identify causal synaptic connections. To assess the feasibility of mapping function onto structure we used a network model that showed a diversity of multineuronal activity patterns and replicated experimental constraints on data acquisition. Using an iterative Bayesian inference algorithm, we detected a select subset of monosynaptic connections substantially more precisely than correlation-based inference, a common alternative approach. We found that precise inference of synaptic connections improved with increasing numbers of diverse multineuronal activity patterns in contrast to increased observations of a single pattern. Surprisingly, neuronal spiking was most effective and precise at revealing causal synaptic connectivity when the lags considered by the iterative Bayesian algorithm encompassed the timescale of synaptic conductance and integration (∼10 ms), rather than synaptic transmission time (∼2 ms), highlighting the importance of synaptic integration in driving postsynaptic spiking. Last, strong synaptic connections were detected preferentially, underscoring their special importance in cortical computation. Even after simulating experimental constraints, top down approaches to cortical connectivity, from function to structure, identify synaptic connections underlying multineuronal activity. These select connections are closely tied to cortical processing.
Functional connectivity in raphé-pontomedullary circuits supports active suppression of breathing during hypocapnic apnea
Authors: Nuding Sarah C. , Segers Lauren S. , Iceman Kimberly E. , O'Connor Russell , Dean Jay B. , Bolser Donald C. , Baekey David M. , Dick Thomas E. , Shannon Roger , Morris Kendall F. , Lindsey Bruce G.
Published: July 2015

Hyperventilation is a common feature of disordered breathing. Apnea ensues if CO2drive is sufficiently reduced. We tested the hypothesis that medullary raphé, ventral respiratory column (VRC), and pontine neurons have functional connectivity and persistent or evoked activities appropriate for roles in the suppression of drive and rhythm during hyperventilation and apnea. Phrenic nerve activity, arterial blood pressure, end-tidal CO2, and other parameters were monitored in 10 decerebrate, vagotomized, neuromuscularly-blocked, and artificially ventilated cats. Multielectrode arrays recorded spiking activity of 649 neurons. Loss and return of rhythmic activity during passive hyperventilation to apnea were identified with the S-transform. Diverse fluctuating activity patterns were recorded in the raphé-pontomedullary respiratory network during the transition to hypocapnic apnea. The firing rates of 160 neurons increased during apnea; the rates of 241 others decreased or stopped. VRC inspiratory neurons were usually the last to cease firing or lose rhythmic activity during the transition to apnea. Mayer wave-related oscillations (0.04–0.1 Hz) in firing rate were also disrupted during apnea. Four-hundred neurons (62%) were elements of pairs with at least one hyperventilation-responsive neuron and a correlational signature of interaction identified by cross-correlation or gravitational clustering. Our results support a model with distinct groups of chemoresponsive raphé neurons contributing to hypocapnic apnea through parallel processes that incorporate disfacilitation and active inhibition of inspiratory motor drive by expiratory neurons. During apnea, carotid chemoreceptors can evoke rhythm reemergence and an inspiratory shift in the balance of reciprocal inhibition via suppression of ongoing tonic expiratory neuron activity.
Warfarin Blocks Gas6-Mediated Axl Activation Required for Pancreatic Cancer Epithelial Plasticity and Metastasis
Authors: Kirane Amanda , Ludwig Kathleen F. , Sorrelle Noah , Haaland Gry , Sandal Tone , Ranaweera Renate , Toombs Jason E. , Wang Miao , Dineen Sean P. , Micklem David , Dellinger Michael T. , Lorens James B. , Brekken Rolf A.
Published: July 2015

Repurposing “old” drugs can facilitate rapid clinical translation but necessitates novel mechanistic insight. Warfarin, a vitamin K “antagonist” used clinically for the prevention of thrombosis for more than 50 years, has been shown to have anticancer effects. We hypothesized that the molecular mechanism underlying its antitumor activity is unrelated to its effect on coagulation, but is due to inhibition of the Axl receptor tyrosine kinase on tumor cells. Activation of Axl by its ligand Gas6, a vitamin K-dependent protein, is inhibited at doses of warfarin that do not affect coagulation. Here, we show that inhibiting Gas6-dependent Axl activation with low-dose warfarin, or with other tumor-specific Axl-targeting agents, blocks the progression and spread of pancreatic cancer. Warfarin also inhibited Axl-dependent tumor cell migration, invasiveness, and proliferation while increasing apoptosis and sensitivity to chemotherapy. We conclude that Gas6-induced Axl signaling is a critical driver of pancreatic cancer progression and its inhibition with low-dose warfarin or other Axl-targeting agents may improve outcome in patients with Axl-expressing tumors. Cancer Res; 75(18); 3699–705. ©2015 AACR.
Mast Cells Infiltrating Inflamed or Transformed Gut Alternatively Sustain Mucosal Healing or Tumor Growth
Authors: Rigoni Alice , Bongiovanni Lucia , Burocchi Alessia , Sangaletti Sabina , Danelli Luca , Guarnotta Carla , Lewis Amy , Rizzo Aroldo , Silver Andrew R. , Tripodo Claudio , Colombo Mario P.
Published: July 2015

Mast cells (MC) are immune cells located next to the intestinal epithelium with regulatory function in maintaining the homeostasis of the mucosal barrier. We have investigated MC activities in colon inflammation and cancer in mice either wild-type (WT) or MC-deficient (KitW-sh) reconstituted or not with bone marrow-derived MCs. Colitis was chemically induced with dextran sodium sulfate (DSS). Tumors were induced by administering azoxymethane (AOM) intraperitoneally before DSS. Following DSS withdrawal, KitW-sh mice showed reduced weight gain and impaired tissue repair compared with their WT littermates or KitW-sh mice reconstituted with bone marrow-derived MCs. MCs were localized in areas of mucosal healing rather than damaged areas where they degraded IL33, an alarmin released by epithelial cells during tissue damage. KitW-sh mice reconstituted with MC deficient for mouse mast cell protease 4 did not restore normal mucosal healing or reduce efficiently inflammation after DSS withdrawal. In contrast with MCs recruited during inflammation-associated wound healing, MCs adjacent to transformed epithelial cells acquired a protumorigenic profile. In AOM- and DSS-treated WT mice, high MC density correlated with high-grade carcinomas. In similarly treated KitW-sh mice, tumors were less extended and displayed lower histologic grade. Our results indicate that the interaction of MCs with epithelial cells is dependent on the inflammatory stage, and on the activation of the tissue repair program. Selective targeting of MCs for prevention or treatment of inflammation-associated colon cancer should be timely pondered to allow tissue repair at premalignant stages or to reduce aggressiveness at the tumor stage. Cancer Res; 75(18); 3760–70. ©2015 AACR.
Cyclin-Dependent Kinase Inhibitor AT7519 as a Potential Drug for MYCN-Dependent Neuroblastoma
Authors: Dolman M. Emmy M. , Poon Evon , Ebus Marli E. , den Hartog Ilona J.M. , van Noesel Carel J.M. , Jamin Yann , Hallsworth Albert , Robinson Simon P. , Petrie Kevin , Sparidans Rolf W. , Kok Robbert J. , Versteeg Rogier , Caron Huib N. , Chesler Louis , Molenaar Jan J.
Published: July 2015

Purpose: MYCN-dependent neuroblastomas have low cure rates with current multimodal treatment regimens and novel therapeutic drugs are therefore urgently needed. In previous preclinical studies, we have shown that targeted inhibition of cyclin-dependent kinase 2 (CDK2) resulted in specific killing of MYCN-amplified neuroblastoma cells. This study describes the in vivo preclinical evaluation of the CDK inhibitor AT7519.

Experimental Design: Preclinical drug testing was performed using a panel of MYCN-amplified and MYCN single copy neuroblastoma cell lines and different MYCN-dependent mouse models of neuroblastoma.

Results: AT7519 killed MYCN-amplified neuroblastoma cell lines more potently than MYCN single copy cell lines with a median LC50 value of 1.7 compared to 8.1 μmol/L (P = 0.0053) and a significantly stronger induction of apoptosis. Preclinical studies in female NMRI homozygous (nu/nu) mice with neuroblastoma patient-derived MYCN-amplified AMC711T xenografts revealed dose-dependent growth inhibition, which correlated with intratumoral AT7519 levels. CDK2 target inhibition by AT7519 was confirmed by significant reductions in levels of phosphorylated retinoblastoma (p-Rb) and nucleophosmin (p-NPM). AT7519 treatment of Th-MYCN transgenic mice resulted in improved survival and clinically significant tumor regression (average tumor size reduction of 86% at day 7 after treatment initiation). The improved efficacy of AT7519 observed in Th-MYCN mice correlated with higher tumor exposure to the drug.

Conclusions: This study strongly suggests that AT7519 is a promising drug for the treatment of high-risk neuroblastoma patients with MYCN amplification. Clin Cancer Res; 21(22); 5100–9. ©2015 AACR.
Multiple Gastrointestinal Polyps in Patients Treated with BRAF Inhibitors
Authors: Amaravadi Ravi K. , Hamilton Kathryn E. , Ma Xiaohong , Piao Shengfu , Portillo Armando Del , Nathanson Katherine L. , Carlino Matteo S. , Long Georgina V. , Puzanov Igor , Xu Xiaowei , Morrissette Jennifer J.D. , Tsai Kenneth Y. , Flaherty Keith T. , Sosman Jeffrey A. , Goodman Grant R. , McArthur Grant A. , Rustgi Anil K. , Metz David C. , Schuchter Lynn M. , Chapman Paul B. , Sepulveda Antonia R.
Published: July 2015

Purpose: BRAF inhibitors (BRAFi) extend survival in BRAF-mutant melanoma but can promote the growth of Ras-mutant neoplasms. This study determined if gastrointestinal polyps found in BRAFi-treated patients harbored Ras mutations.

Experimental Design: Colonic and gastric polyps were identified and resected from BRAFi-treated melanoma patients. Next-generation sequencing (NGS) was performed on polyps. The ability of BRAFi to promote polyp formation was functionally characterized in Apc Min+/− mice. MAPK and β-catenin pathway activity was assessed by immunohistochemistry in mouse and human polyps.

Results: Fourteen patients treated with BRAFi underwent endoscopy to assess for polyps. Seven out of 7 patients >40 years of age and treated for >2 years were found to have colonic tubular adenomas with 4 out of the 7 patients having 5 or more polyps. One patient presented with bleeding from hyperplastic gastric polyps that recurred 6 months after BRAFi rechallenge. NGS performed on polyps found no mutations in MAPK pathway genes, but found APC mutations in all tubular adenomas. A significant increase in the number of polyps was observed in BRAFi-treated compared with control-treated Apc Min+/− mice (20.8 ± 9.2 vs 12.8 ± 0.1; P = 0.016). No polyps were observed in BRAFi-treated wild-type mice.

Conclusions: BRAFi may increase the risk of developing hyperplastic gastric polyps and colonic adenomatous polyps. Due to the risk of gastrointestinal bleeding and the possibility of malignant transformation, further studies are needed to determine whether or not endoscopic surveillance should be recommended for patients treated with BRAFi. Clin Cancer Res; 21(23); 5215–21. ©2015 AACR.
EGFR Exon 18 Mutations in Lung Cancer: Molecular Predictors of Augmented Sensitivity to Afatinib or Neratinib as Compared with First- or Third-Generation TKIs
Authors: Kobayashi Yoshihisa , Togashi Yosuke , Yatabe Yasushi , Mizuuchi Hiroshi , Jangchul Park , Kondo Chiaki , Shimoji Masaki , Sato Katsuaki , Suda Kenichi , Tomizawa Kenji , Takemoto Toshiki , Hida Toyoaki , Nishio Kazuto , Mitsudomi Tetsuya
Published: July 2015

Purpose: Lung cancers harboring common EGFR mutations respond to EGFR tyrosine kinase inhibitors (TKI), whereas exon 20 insertions (Ins20) are resistant to them. However, little is known about mutations in exon 18.

Experimental Design: Mutational status of lung cancers between 2001 and 2015 was reviewed. Three representative mutations in exon 18, G719A, E709K, and exon 18 deletion (Del18: delE709_T710insD) were retrovirally introduced into Ba/F3 and NIH/3T3 cells. The 90% inhibitory concentrations (IC90s) of first-generation (1G; gefitinib and erlotinib), second-generation (2G; afatinib, dacomitinib, and neratinib), and third-generation TKIs (3G; AZD9291 and CO1686) were determined.

Results: Among 1,402 EGFR mutations, Del19, L858R, and Ins20 were detected in 40%, 47%, and 4%, respectively. Exon 18 mutations, including G719X, E709X, and Del18, were present in 3.2%. Transfected Ba/F3 cells grew in the absence of IL3, and NIH/3T3 cells formed foci with marked pile-up, indicating their oncogenic abilities. IC90s of 1G and 3G TKIs in G719A, E709K, and Del18 were much higher than those in Del19 (by >11–50-fold), whereas IC90s of afatinib were only 3- to 7-fold greater than those for Del19. Notably, cells transfected with G719A and E709K exhibited higher sensitivity to neratinib (by 5–25-fold) than those expressing Del19. Patients with lung cancers harboring G719X exhibited higher response rate to afatinib or neratinib (∼80%) than to 1G TKIs (35%–56%) by compilation of data in the literature.

Conclusions: Lung cancers harboring exon 18 mutations should not be overlooked in clinical practice. These cases can be best treated with afatinib or neratinib, although the currently available in vitro diagnostic kits cannot detect all exon 18 mutations. Clin Cancer Res; 21(23); 5305–13. ©2015 AACR.
“Braking” the Cycle of Resistance in Endocrine Therapy for Breast Cancer
Authors: DeMichele Angela , Chodosh Lewis A.
Published: July 2015

Endocrine resistance leads to recurrence and death from breast cancer. Animal models of endocrine resistance enable preclinical identification of efficacious therapeutic combinations and further our understanding of resistance. This strategy provides new insights into optimally targeting interactions between estrogen receptor (ESR-1) activity and the cell cycle by CDK4/6 inhibitors. Clin Cancer Res; 21(22); 4999–5001. ©2015 AACR.

See related article by Wardell et al., p. 5121
NSCLC Driven by DDR2 Mutation Is Sensitive to Dasatinib and JQ1 Combination Therapy
Authors: Xu Chunxiao , Buczkowski Kevin A. , Zhang Yanxi , Asahina Hajime , Beauchamp Ellen M. , Terai Hideki , Li Yvonne Y. , Meyerson Matthew , Wong Kwok-kin , Hammerman Peter S.
Published: July 2015

Genetically engineered mouse models of lung cancer have demonstrated an important role in understanding the function of novel lung cancer oncogenes and tumor-suppressor genes identified in genomic studies of human lung cancer. Furthermore, these models are important platforms for preclinical therapeutic studies. Here, we generated a mouse model of lung adenocarcinoma driven by mutation of the discoidin domain receptor 2 (DDR2) gene combined with loss of TP53. DDR2L63V;TP53L/L mice developed poorly differentiated lung adenocarcinomas in all transgenic animals analyzed with a latency of 40 to 50 weeks and a median survival of 67.5 weeks. Mice expressing wild-type DDR2 with combined TP53 loss did not form lung cancers. DDR2L63V;TP53L/L tumors displayed robust expression of DDR2 and immunohistochemical markers of lung adenocarcinoma comparable with previously generated models, though also displayed concomitant expression of the squamous cell markers p63 and SOX2. Tumor-derived cell lines were not solely DDR2 dependent and displayed upregulation of and partial dependence on MYCN. Combined treatment with the multitargeted DDR2 inhibitor dasatinib and BET inhibitor JQ1 inhibited tumor growth in vitro and in vivo. Together, these results suggest that DDR2 mutation can drive lung cancer initiation in vivo and provide a novel mouse model for lung cancer therapeutics studies. Mol Cancer Ther; 14(10); 2382–9. ©2015 AACR.
Regulatory T-cell Response to EnterotoxigenicBacteroides fragilisColonization Triggers IL17-Dependent Colon Carcinogenesis
Authors: Geis Abby L. , Fan Hongni , Wu Xinqun , Wu Shaoguang , Huso David L. , Wolfe Jaime L. , Sears Cynthia L. , Pardoll Drew M. , Housseau Franck
Published: July 2015

Many epithelial cancers are associated with chronic inflammation. However, the features of inflammation that are procarcinogenic are not fully understood. Regulatory T cells (Treg) typically restrain overt inflammatory responses and maintain intestinal immune homeostasis. Their immune-suppressive activity can inhibit inflammation-associated cancers. Paradoxically, we show that colonic Tregs initiate IL17-mediated carcinogenesis in multiple intestinal neoplasia mice colonized with the human symbiote enterotoxigenic Bacteroides fragilis (ETBF). Depletion of Tregs in ETBF-colonized C57BL/6 FOXP3DTR mice enhanced colitis but diminished tumorigenesis associated with shifting of mucosal cytokine profile from IL17 to IFNγ; inhibition of ETBF-induced colon tumorigenesis was dependent on reduced IL17 inflammation and was independent of IFNγ. Treg enhancement of IL17 production is cell-extrinsic. IL2 blockade restored Th17 responses and tumor formation in Treg-depleted animals. Our findings demonstrate that Tregs limit the availability of IL2 in the local microenvironment, allowing the Th17 development necessary to promote ETBF-triggered neoplasia, and thus unveil a new mechanism whereby Treg responses to intestinal bacterial infection can promote tumorigenesis.

Significance: Tregs promote an oncogenic immune response to a common human symbiote associated with inflammatory bowel disease and colorectal cancer. Our data define mechanisms by which mucosal Tregs, despite suppressing excessive inflammation, promote the earliest stages of immune procarcinogenesis via enhancement of IL17 production at the expense of IFNγ production. Cancer Discov; 5(10); 1098–109. ©2015 AACR.

See related commentary by Irrazabal and Martin, p. 1021.

This article is highlighted in the In This Issue feature, p. 1005
Development of Suck and Swallow Mechanisms in Infants
Authors: Lau Chantal
Published: July 2015

Preterm infants' hospital discharge is often delayed due to their inability to feed by mouth safely and competently. No evidence-based supported guidelines are currently available for health-care professionals caring for these infants. Available interventions advocating benefits are not readily acknowledged for lack of rigorous documentation inasmuch as any improvements may ensue from infants' normal maturation. Through research, a growing understanding of the development of nutritive sucking skills has emerged, shedding light on how and why infants may encounter oral feeding difficulties due to the immaturity of specific physiologic functions. Unfortunately, this knowledge has yet to be translated to the clinical practice to improve the diagnoses of oral feeding problems through the development of relevant assessment tools and to enhance infants' oral feeding skills through the development of efficacious preventive and therapeutic interventions. This review focuses on the maturation of the various physiologic functions implicated in the transport of a bolus from the oral cavity to the stomach. Although infants' readiness for oral feeding is deemed attained when suck, swallow, and respiration are coordinated, we do not have a clear definition of what coordination implies. We have learned that each of these functions encompasses a number of elements that mature at different times and rates. Consequently, it would appear that the proper functioning of sucking, the swallow processing, and respiration need to occur at two levels: first, the elements within each function must reach an appropriate functional maturation that can work in synchrony with each other to generate an appropriate suck, swallow process, and respiration; and second, the elements of all these distinct functions, in turn, must be able to do the same at an integrative level to ensure the safe and efficient transport of a bolus from the mouth to the stomach.
Heat Shock Protein 70 and CHIP Promote Nox4 Ubiquitination and Degradation within the Losartan Antioxidative Effect in Proximal Tubule Cells
Authors: Gil Lorenzo Andrea F. , Costantino Valeria V. , Appiolaza Martin López , Cacciamani Valeria , Benardon Maria E. , Bocanegra Victoria , Vallés Patricia G.
Published: July 2015

Background: Angiotensin II/Angiotensin II type 1 receptor (AT1R) effects are dependent on ROS production stimulated by NADPH oxidase activation. Hsp70 regulates a diverse set of signaling pathways through their interactions with proteins. CHIP is a E3 ubiquitin ligase that targets proteins for polyubiquitination and degradation. Aim: We study whether Hsp70/CHIP contribute to the negative regulation of Nox4 after AT1R blockage. Methods/Results: Primary culture of proximal tubule epithelial cells (PTCs) from SHR and WKY were stimulated with Angiotensin II (AII) or treated with Losartan (L) or Losartan plus Angiotensin II (L+AII). Losartan decreased AT1R and Nox4 while enhancing caveolin-1 and Hsp70 protein expression in SHR PTCs. Immunoprecipitation and immunofluorescence proved interaction and colocalization of increased Hsp70/CHIP with decreased Nox4 in SHR PTCs (L) vs (All). Hsp72 knockdown resulted in enhanced Nox4 protein levels, NADPH oxidase activity and ROS generation in (L+AII) revealing that Losartan was unable to abrogate AII effects on Nox4 expression and oxidative activity. Moreover, MG132 exposed PTCs (L) demostrated blocked ubiquitinated Nox4 degradation and increased colocalization of Nox4/Ubiquitin by inmunofluorescence. Conversely, Hsp72 depletion reduced Nox4/Ubiquitin colocalization causing Nox4 upregulation due to proteosomal degradation inhibition, although Losartan treatment. Conclusion: Our study demonstrates that Hsp70 and CHIP mediates the ubiquitination and proteasomal degradation of Nox4 as part of the antioxidative effect of Losartan in SHR.
Impact of p38 MAP Kinase Inhibitors on LPS-Induced Release of TNF-α in Whole Blood and Primary Cells from Different Species
Authors: Fehr Sarah , Unger Anke , Schaeffeler Elke , Herrmann Sonja , Laufer Stefan , Schwab Matthias , Albrecht Wofgang
Published: July 2015

Background/Aims: Inhibition of p38 mitogen-activated protein kinase (p38 MAPK) is promising for the treatment of inflammatory disorders, however, the efficacy of p38 MAPK inhibitors in clinical trials is limited so far. Since functional sensitivity of p38 MAPK is commonly predicted by preclinical species, we systematically investigated interspecies differences including human tissue. Methods: Ex vivo test models were established using whole blood and primary cells from different species such as mice, rats, pigs and humans to compare LPS-induced TNF-α inhibition of four different p38 MAPK reference inhibitors SB 203580, BIRB-796, Pamapimod, and a Losmapimod analogue as well as a proprietary imidazole-based p38 MAPK Inhibitor. Results: All analysed p38 MAPK inhibitors resulted in significant inhibition of LPS-induced TNF-α release but with high interspecies differences for dose sensitivity. IC50 values from human whole blood and PBMC showed significant higher sensitivity towards p38 MAPK inhibition compared with data from pig and rat. Conclusion: Inhibition of TNF-α release by p38 MAPK inhibitors can be reliably identified in well-established laboratory species such as rat or mouse. However, our data indicate that animal models appear to be limited for valid prediction of the inhibitory potential for TNF-α release in humans. Thus, human tissues should be considered early in the drug development process of p38 MAPK inhibitors.
Role of Exercise in the Activation of Brown Adipose Tissue
Authors: Sanchez-Delgado Guillermo , Martinez-Tellez Borja , Olza Josune , Aguilera Concepcion M. , Gil Ángel , Ruiz Jonatan R.
Published: July 2015

Background: The energy-burning capacity of brown adipose tissue (BAT) makes it an attractive target for use in anti-obesity therapies. Moreover, due to its ability to oxidize glucose and lipids, BAT activation has been considered a potential therapy to combat type 2 diabetes and atherogenesis. Summary: BAT is mainly regulated by the sympathetic nervous system (SNS); yet, recent findings have shown a group of novel activators that act independently of the stimulation of the SNS such as cardiac natriuretic peptides, irisin, interleukin-6, β-aminoisobutyric acid and fibroblast growth factor 21 that could influence BAT metabolism. Several strategies are being examined to activate and recruit BAT with no side effects. In this review, we postulate that exercise might activate and recruit human BAT through the activation of SNS, heart and skeletal muscle. Key Messages: Epidemiological and well-designed exercise-based randomized controlled studies are needed to clarify if exercise is able to activate BAT in humans.
Diagnostic Yield of Venous Thrombosis and Pulmonary Embolism by Combined CT Venography and Pulmonary Angiography in Patients with Cryptogenic Stroke and Patent Foramen Ovale
Authors: Lapergue Bertrand , Decroix Jean Pierre , Evrard Serge , Wang Adrien , Bendetowicz David , Offroy Marie Astrid , Graveleau Philippe , Russel Stephanie , Ramdane Nassima , Mellot François , Scherrer Antoine , Bourdain Frédéric
Published: July 2015

Background: Paradoxical embolism via a patent foramen ovale (PFO) has been suggested as a potential stroke mechanism. Combined CT venography and pulmonary angiography (CVPA) is a simple, validated and accurate technique to diagnose deep venous thrombosis (DVT) or pulmonary embolism (PE). We sought to assess the prevalence of DVT or PE among patients with PFO and cryptogenic stroke (CS) by CVPA. Methods: Patients were identified retrospectively from a clinical registry of consecutive patients with stroke admitted to our Stroke Unit. The following criteria were required for inclusion in this study: CS, PFO identified by transthoracic echography using contrast medium and CVPA performed during the hospitalization following stroke. Results: A total of 114 patients with PFO underwent a CVPA within 7 days (interquartile range 4-9) from stroke symptom onset. On cerebral imaging, 11% had multiple infarcts. CVPA documented deep vein thrombosis (DVT) in 10 patients (8.8%) and PE in 5 patients (4.4%), that is, a total of 12 patients with prevalence of 10.5% (95% CI 5.5-17.7). Patients with PE-DVT had higher D-dimers and C reactive protein level than patients without PE-DVT (p < 0.05). Conclusion: CVPA may be used by the stroke team in the work-up of suspected paradoxical embolism among cryptogenic ischemic stroke patients with PFO.
Psoriasiform Drug Eruption Caused by Abatacept: Immunohistochemical Investigation of STAT Signaling
Authors: Tanita Kayo , Fujimura Taku , Kakizaki Aya , Furudate Sadanori , Kusakari Yoshiyuki , Aiba Setsuya
Published: July 2015

Abatacept is a biological immune modifier that is used for the treatment of rheumatoid arthritis. Although psoriasiform drug eruption is reported as one of the cutaneous adverse effects of abatacept, the precise mechanisms are not fully understood. In this report, we describe a 65-year-old Japanese man with psoriasiform drug eruption caused by abatacept. Interestingly, immunohistochemical staining revealed that the epidermal keratinocytes in the basal layer and lower layers of the stratum spinosum were positive for pSTAT3, partially positive for pSTAT1 and negative for pSTAT6, which is similar to conventional psoriasis vulgaris. Our present study suggests that psoriasiform drug eruption caused by abatacept might develop by similar immunological mechanisms as those of psoriasis vulgaris.
Low Bone Turnover in Chronic Kidney Disease Is Associated with Decreased VEGF-A Expression and Osteoblast Differentiation
Authors: Chen Neal X. , O''Neill Kalisha D. , Allen Matthew R. , Newman Christopher L. , Moe Sharon M.
Published: July 2015

Background: Low turnover bone (low bone formation rates (BFRs)) with decreased osteoblast number is common in patients with chronic kidney disease (CKD) and attributed to ‘over-suppression' of the parathyroid hormone (PTH) despite supra-physiologic levels. An alternative hypothesis is abnormal osteoblast differentiation, resulting in low BFRs due to reduced VEGF-A. Methods: We analyzed the expression of VEGF-A and mesenchymal stem cell (MSC) differentiation factors in freshly isolated bone marrow (BM) cells, and in BM cell-derived MSC in rats with different levels of BFRs and PTH (modulated by calcium and zoledronic acid). The regulators of VEGF in MSC were also determined. Results: VEGF-A expression was reduced in the BM cells from CKD vs. normal animals (p < 0.02). In BM-derived MSC from CKD, there were decreased osteoblast transcription factors and mineralization. In CKD animals, the BM VEGF-A expression was positively correlated with BFR (r = 0.80, p < 0.001). Reducing BFRs in CKD animals led to reductions in VEGF-A expression and osteoblast transcription factors regardless of the PTH level. We therefore examined other regulators of VEGF-A and found decreased expression of hypoxia-inducible factor-1α and the master transcription factor of antioxidants nuclear factor (erythroid-derived 2)-like 2 in CKD animals with low PTH. Conclusion: Low BFRs in CKD are associated with a basal decrease in VEGF-A expression in BM that may be driven by altered hypoxia and oxidative stress.
The Role of Fludrocortisone in Cognition and Mood in Patients with Primary Adrenal Insufficiency (Addison's Disease)
Authors: Schultebraucks Katharina , Wingenfeld Katja , Otte Christian , Quinkler Marcus
Published: July 2015

Background: Primary adrenal insufficiency (AI) requires hormone replacement therapy with fludrocortisone and hydrocortisone stimulating glucocorticoid (GR) and mineralocorticoid receptors (MR). Evidence from animal and human studies shows that MR function is crucial for cognitive function and mood. Regarding patients with AI, very little is known about the role of MR in cognitive function and mood. Methods: A repeated-measures within-subject design was used to determine whether cognitive function and mood are related to MR occupation in patients with AI. Intraindividually, patients were examined twice, with 1 week between testing days: once with fludrocortisone (high MR occupation) and once without fludrocortisone (low MR occupation). All patients kept their stable regimen of hydrocortisone. The assessment of cognitive function included executive function, attention, and verbal, visuospatial and working memory. Additionally, mood and blood pressure were measured. Results: Verbal memory improved significantly during high MR occupation (after fludrocortisone intake) compared to low MR occupation [without fludrocortisone, t(29) = -2.1, p = 0.046]. There were trend level differences in the Number-Combination test [t(29) = -1.9, p = 0.074] and in the Stroop interference task [t(29) = -1.9, p = 0.068]. No significant differences in visuospatial and working memory were found. Furthermore, the current mood state was better during high MR occupation compared to low MR occupation [t(29) = -2.4, p = 0.023] as was diastolic blood pressure [F(2, 29) = 3.6, p = 0.07]. Conclusions: Cognitive function and mood in patients with AI depend in part on MR occupation. Because the medium effect size indicates a potential clinical significance, further studies should systematically examine which dosages of fludrocortisone are associated with optimal cognitive function and mood in AI patients.
HMGB1 binds to activated platelets via the receptor for advanced glycation end products and is present in platelet rich human coronary artery thrombi
Authors: Bode Michael , Haenel David , Hagemeyer Christoph E. , Seeba Hannah , Duerschmied Daniel , Bassler Nicole , Jandeleit-Dahm Karin A. , Sweet Matthew J. , Agrotis Alex , Bobik Alex , Peter Karlheinz , Ahrens Ingo , Chen Yung-Chih , Topcic Danijal
Published: July 2015

High mobility group box 1 (HMGB1) acts as both a nuclear protein that regulates gene expression, as well as a pro-inflammatory alarmin that is released from necrotic or activated cells. Recently, HMGB1-expression in human atherosclerotic plaques was identified. Therapeutic blockade of HMGB1 reduced the development of diet-induced atherosclerosis in ApoE knockout mice. Thus, we hypothesised an interaction between HMGB1 and activated platelets. Binding of recombinant HMGB1 to platelets was assessed by flow cytometry. HMGB1 bound to thrombin-activated human platelets (MFI 2.49 vs 25.01, p=0.0079). Blood from wild-type, TLR4 and RAGE knockout mice was used to determine potential HMGB1 receptors on platelets. HMGB1 bound to platelets from wild type C57Bl6 (MFI 2.64 vs 20.3, p< 0.05), and TLR4-/- mice (MFI 2.11 vs 25.65, p< 0.05) but failed to show binding to platelets from RAGE-/- mice (p > 0.05). RAGE expression on human platelets was detected by RT-PCR with mRNA extracted from highly purified platelets and confirmed by Western blot and immunofluorescence microscopy. Platelet activation increased RAGE surface expression (MFI 4.85 vs 6.74, p< 0.05). Expression of HMGB1 in human coronary artery thrombi was demonstrated by immunohistochemistry and revealed high expression levels. Platelets bind HMGB1 upon thrombin-induced activation. Platelet specific expression of RAGE could be detected at the mRNA and protein level and is involved in the binding of HMGB1. Furthermore, platelet activation up-regulates platelet surface expression of RAGE. HMGB1 is highly expressed in platelet-rich human coronary artery thrombi pointing towards a central role for HMGB1 in atherothrombosis, thereby suggesting the possibility of platelet targeted anti-inflammatory therapies for atherothrombosis.
Extracorporeal Membrane Oxygenation as a Bridge to Pediatric Heart Transplantation
Authors: Dipchand Anne I. , Mahle William T. , Tresler Margaret , Naftel David C. , Almond Christopher , Kirklin James K. , Pruitt Elizabeth , Webber Steven A.
Published: July 2015

Background—

Current organ allocation algorithms direct hearts to the sickest recipients to mitigate death while waiting. This may result in lower post-transplant (Tx) survival for high-risk candidates mandating close examination to determine the appropriateness of different technologies as a bridge to Tx.

Methods and Results—

We analyzed all patients (<18 years old) from the Pediatric Heart Transplant Study (PHTS) database listed for heart Tx (1993–2013) to determine the effect of extracorporeal membrane oxygenation (ECMO) support at the time of listing and the time of Tx on waitlist mortality and post-Tx outcomes. Eight percent of patients were listed on ECMO, and within 12 months, 49% had undergone Tx, 35% were deceased, and 16% were alive waiting. Survival at 12 months after listing (censored at Tx) was worse in patients on ECMO at listing (50%) compared with ventricular assist device at listing (76%) or not on ECMO or ventricular assist device at listing (76%;
P
<0.0001). Two hundred three (5%) patients underwent Tx from ECMO; 135 (67%) had been on ECMO since listing, and 67 (33%) had deteriorated to ECMO support while waiting. Survival after Tx was worse in patients who underwent Tx from ECMO (3 years: 64%) versus on ventricular assist device at Tx (3 years: 84%) or not on ECMO/ventricular assist device at Tx (3 years: 85%;
P
<0.0001). Patients transplanted from ECMO at age <1 year had the worst survival.

Conclusions—

Pediatric patients requiring ECMO support before heart Tx have poor outcomes. Prioritization of donor hearts to children waitlisted on ECMO warrants careful consideration because of ECMO’s high pre- and post-Tx mortality.
Residual Risk of Stroke and Death in Anticoagulated Patients According to the Type of Atrial Fibrillation
Authors: Senoo Keitaro , Lip Gregory Y.H. , Lane Deirdre A. , Büller Harry R. , Kotecha Dipak
Published: July 2015

Background and Purpose—

Atrial fibrillation (AF) and heart failure frequently coexist and are associated with increased morbidity and mortality. We investigated the prognosis of anticoagulated patients with permanent AF and nonpermanent AF according to preexisting heart failure in the AMADEUS (Evaluating the Use of SR34006 Compared to Warfarin or Acenocoumarol in Patients With Atrial Fibrillation) trial.

Methods—

The primary outcome was a composite of cardiovascular death and stroke or systemic embolism, analyzed using a Cox proportional hazards model, adjusted for baseline age, sex, diabetes mellitus, hypertension, creatinine, and previous cardiovascular diseases. The median follow-up was 11.6 months (interquartile range, 6.2–15.2).

Results—

Nonpermanent AF was present in 2072 patients (46% of cohort), of which 339 (16%) had preexisting heart failure. A total of 2484 patients had permanent AF (54% of cohort), with a higher burden of heart failure including 730 patients (29%;
P
<0.001). Overall, death because of cardiovascular causes occurred in 57 patients and 45 had stroke or systemic embolism (1.4/100 person-years for each). Overall, the adjusted incidence of the composite outcome was higher in patients with permanent AF than in patients with nonpermanent AF. In multivariate analysis, permanency of AF, creatinine, prior cerebrovascular events, and previous coronary disease were independently associated with the primary outcome. The hazard ratio for permanent versus nonpermanent AF was 1.68 (95% confidence interval, 1.08–2.55;
P
=0.02). The presence of heart failure increased the risk of adverse outcomes in a similar way in both permanent and nonpermanent AF (interaction
P
value=0.76).

Conclusions—

The risk of cardiovascular death, stroke, or systemic embolism is higher in anticoagulated patients with permanent AF than in those with nonpermanent AF, regardless of preexisting heart failure.
Prevalence of Individuals Experiencing the Effects of Stroke in Canada
Authors: Krueger Hans , Koot Jacqueline , Hall Ruth E. , O’Callaghan Christina , Bayley Mark , Corbett Dale
Published: July 2015

Background and Purpose—

Previous estimates of the number and prevalence of individuals experiencing the effects of stroke in Canada are out of date and exclude critical population groups. It is essential to have complete data that report on stroke disability for monitoring and planning purposes. The objective was to provide an updated estimate of the number of individuals experiencing the effects of stroke in Canada (and its regions), trending since 2000 and forecasted prevalence to 2038.

Methods—

The prevalence, trends, and projected number of individuals experiencing the effects of stroke were estimated using region-specific survey data and adjusted to account for children aged <12 years and individuals living in homes for the aged.

Results—

In 2013, we estimate that there were 405 000 individuals experiencing the effects of stroke in Canada, yielding a prevalence of 1.15%. This value is expected to increase to between 654 000 and 726 000 by 2038. Trends in stroke data between 2000 and 2012 suggest a nonsignificant decrease in stroke prevalence, but a substantial and rising increase in the number of individuals experiencing the effects of stroke. Stroke prevalence varied considerably between regions.

Conclusions—

Previous estimates of stroke prevalence have underestimated the true number of individuals experiencing the effects of stroke in Canada. Furthermore, the projected increases that will result from population growth and demographic changes highlight the importance of maintaining up-to-date estimates.
Putin’s Elite and the Legacies of Soviet Quasi-Modernisation in Contemporary Russia
Authors: Pakulski Jan
Published: July 2015

In spite of its rich energy resources and strategic location, and in spite of the declared commitment to further liberal reforms by politi-cal leaders, Russia seems to be experiencing some transition and de-velopmental problems. Economic growth is uneven and relatively slow (compared with other BRICS and oil/gas exporters). The Russian state remains centralistic, controlling, corrupt and inert. Russian politics is undergoing a retrogressive ‘authoritarian turn’ accompanied by a tightening of control over the media. The political competition for the top political offices is so skewed, the opposition so restricted, and the mass media so constrained, that the Freedom House no longer classi-fies Russia as ‘free’ and ‘democratic’. State patronage, protectionism and the taxation burden remain high, corruption is endemic, access to financing is limited – all constraining Russia’s development and pre-venting the advanced modernisation of the state, economy and society. These developments in contemporary Russia are analysed and ex-plained as correlates of the outlooks, strategic goals and power inter-ests of the new Russian elite – Putin’s siloviki. The developmental problems of contemporary Russia are linked to the ‘Soviet legacies’ of socialist ‘quasi-modernisation’ revived by Putin’s siloviki in their at-tempt to consolidate power and stabilise politics by ‘restoring’ Rus-sia’s power and influence.
Beyond the Sneer: Revisiting Musical Socialist Realism
Authors: Crotty Joel
Published: July 2015

The author engages with how communism and socialist realism have been re-presented in post-communist Europe. Communist cultural artefacts are displayed in museums and theme parks on a continuum from dangerous relics to benign kitsch. He argues for neutrality when dealing with musical socialist realism and demonstrates some of the pitfalls in taking that position through examples of Romanian art music from the mid-20th century.
An Unknown Australian War Novel: Miles Franklin’s Nemari ništa: Six Months with the Serbs
Authors: Vladiv-Glover Slobodanka
Published: July 2015

Miles Franklin’s sketches from the Balkan front are discussed in the context of Franklin as an Australian writer of “the soil” whose observations of the Serbian soldiers and life in the hospital camp on the Salonika front take the form of sketches of manners. The sketch of manners is announced in the literary manifestoes of the French, the English and the Russians as part of a poetics of Realism in the European literary canon. These prescribe the capturing of a moment in time as the task of the writer, who acts as a ‘local historian’ perpetuating the memory of the manners and mores of his contemporaries for posterity. Franklin’s sketches comply also with the idea of ‘circumstantial beauty’ propounded by Charles Baudelaire in his essay on Constantin Guys. It is argued that the Serbs portrayed by Franklin become a symbol of the desire of the age. The sketches transcend the modest claims of the writer of being simple testimony to the Serb as she saw him and are a chronicle of feelings of the author, her reactions to sounds, sights and the humanity she encounters. It is the personalized emotive point of view, described as “Australianism” which imparts unity to the sketches and gives grounds for calling Six Months with the Serbs an Australian war novel.
Introduction
Authors: Sujecka Jolanta
Published: July 2015

IntroductionThe text presents the leading subject of the issue, and discusses the question of cultural dis-contiuity, with special interest to its Balkan context.