bluuebunny/crossref_metadata_2025 huggingface dataset:
INTERACTION OF PULSATILE FLOW WITH PERISTALTIC TRANSPORT OF A VISCOELASTIC FLUID: CASE OF A MAXWELL FLUID
Authors: RACHID H. , OUAZZANI M. T.
Published: September 2014

This article analytically investigates the interaction of pulsatile flow with peristaltic transport of a viscoelastic Maxwell fluid in a cylindrical tube. The flow is considered unsteady even in the wave frame analysis where we impose a periodic pressure gradient. This transport is studied under low Reynolds number and long wavelength approximations. The governing equations are developed up to the second-order in the Deborah number and the Womersley number. We first analyzed the impact of the pulsatile flow, of the occlusion and of the viscoelastic effects of fluid on the pressure rise and on the friction force. Physical behavior of different parameters of the problem has been graphically presented and the influence of these parameters on the mechanical efficiency has been analyzed.
Chapter 18 East Antarctic continental shelf: Prydz Bay and the Mac.Robertson Land Shelf
Authors: O'Brien P. E. , Harris P. T. , Post A. L. , Young N.
Published: September 2014

Prydz Bay and the Mac.Robertson Land Shelf exhibit many of the variations seen on Antarctic continental shelves. The Mac.Robertson shelf is relatively narrow with rugged, inner-shelf topography and shallow outer-shelf banks swept by the west-flowing Antarctic Coastal Current. U-shaped valleys cut across the shelf. It has thin sedimentary cover, deposited and eroded by cycles of glacial advance and retreat through the Neogene and Quaternary. Modern sedimentation is diatom-rich siliceous, muddy ooze in shelf deeps, while, on the banks, phytodetritus, calcareous bioclasts and terrigenous material are mixed by iceberg ploughing. Prydz Bay is a large embayment fed by the Amery Ice Shelf. It has a broad inner-shelf deep area and outer bank, with depths ranging from 2400 m beneath the ice shelf to 100 m on the outer banks. A clockwise gyre flows through the bay. Fine mud and siliceous ooze drape the seafloor; however, banks are scoured by icebergs to depths as great as 500 m. The Mac.Robertson shelf has seen advances to the shelf edge during glacial episodes and retreat during warming and rising sea level. Prydz Bay shows more complexity, with parts of the bay showing partial advance of the ice-grounding zone.
To the islands born: the research potential of submerged landscapes and human habitation sites from the islands of NW Australia
Authors: Ward I. , Veth P. , Manne T.
Published: September 2014

This paper focuses on the continental shelf of NW Australia, and on models for change in littoral and offshore environments of relevance to human occupation over the last 50 kyr. Major island groups occur on the shelf including the Montebello and Barrow islands, and those of the Dampier Archipelago. At lowest sea level around 22 ka, these would have been uplands that then became progressively isolated by subsequent sea-level rise. By integrating archaeological and zooarchaeological records from excavations on these islands with the geology and geomorphology, we interpret palaeoeconomic resource potential in relation to changing sea level and coastline morphology. The preservation potential for submerged archaeological sites and features is also assessed. Current archaeological evidence from these offshore islands indicates that the submerged coastal landscape is likely to have been a potentially rich environment for resources and human occupation, even at times of lowest sea level and regional aridity. Should any exploration of submerged archaeology be carried out in this region, it is likely to be rewarding, offering unique insights into Late Pleistocene coastal occupation.
Marine geological and archaeological evidence of a possible pre-Neolithic site in Pantelleria Island, Central Mediterranean Sea
Authors: Abelli Leonardo , Agosto Maria Vittoria , Casalbore Daniele , Romagnoli Claudia , Bosman Alessandro , Antonioli Fabrizio , Pierdomenico Martina , Sposato Andrea , Chiocci Francesco Latino
Published: September 2014

Recent underwater archaeological surveys recovered hundreds of flint artefacts between depths of 18 and 21 m at Cala Tramontana, a small bay located in the eastern part of Pantelleria Island. Most of the flint artefacts indicate debitage, and are characterized by cores and flakes without any specific morphology. Different lithic tools were also identified, such as fragments of blades, truncations, end-scrapers, points and crested blades. An initial hypothesis is that this lithic industry represents the oldest traces of human visitation to the island, possibly related to the exploitation of the nearby obsidian source, and favoured because of the sheltered coastal configuration of Cala Tramontana and Cala Levante with respect to the dominant winds and related storms. However, the present-day coastal setting in the bay is rather inhospitable, with high cliffs and difficult marine access. In contrast, palaeo-landscape reconstructions by means of high-resolution multibeam bathymetry reveal the possible presence of a small palaeo-beach in the inner part of the bay when the sea level was 15 m lower than at present. By comparing this palaeo-sea level with the eustatic curve (and by excluding possible vertical movements), we roughly estimate an age of the lithic industry of 9.6–7.7 cal ka BP.
Acute consumption of a high-fat diet prior to ischemia-reperfusion results in cardioprotection through NF-κB-dependent regulation of autophagic pathways
Authors: Haar Lauren , Ren Xiaoping , Liu Yong , Koch Sheryl E. , Goines Jillian , Tranter Michael , Engevik Melinda A. , Nieman Michelle , Rubinstein Jack , Jones W. Keith
Published: September 2014

Previous studies have demonstrated improvement of cardiac function occurs with acute consumption of a high-fat diet (HFD) after myocardial infarction (MI). However, no data exist addressing the effects of acute HFD upon the extent of injury after MI. This study investigates the hypothesis that short-term HFD, prior to infarction, protects the heart against ischemia-reperfusion (I/R) injury through NF-κB-dependent regulation of cell death pathways in the heart. Data show that an acute HFD initiates cardioprotection against MI (>50% reduction in infarct size normalized to risk region) after 24 h to 2 wk of HFD, but protection is completely absent after 6 wk of HFD, when mice are reported to develop pathophysiology related to the diet. Furthermore, cardioprotection after 24 h of HFD persists after an additional 24 h of normal chow feeding and was found to be dependent upon NF-κB activation in cardiomyocytes. This study also indicates that short-term HFD activates autophagic processes (beclin-1, LC-3) preischemia, as seen in other protective stimuli. Increases in beclin-1 and LC-3 were found to be NF-κB-dependent, and administration of chloroquine, an inhibitor of autophagy, abrogated cardioprotection. Our results support that acute high-fat feeding mediates cardioprotection against I/R injury associated with a NF-κB-dependent increase in autophagy and reduced apoptosis, as has been found for ischemic preconditioning.
Central SDF-1/CXCL12 expression and its cardiovascular and sympathetic effects: the role of angiotensin II, TNF-α, and MAP kinase signaling
Authors: Wei Shun-Guang , Zhang Zhi-Hua , Yu Yang , Felder Robert B.
Published: September 2014

The chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) and its receptors are expressed by neurons and glial cells in cardiovascular autonomic regions of the brain, including the hypothalamic paraventricular nucleus (PVN), and contribute to neurohumoral excitation in rats with ischemia-induced heart failure. The present study examined factors regulating the expression of SDF-1 in the PVN and mechanisms mediating its sympatho-excitatory effects. In urethane anesthetized rats, a 4-h intracerebroventricular (ICV) infusion of angiotensin II (ANG II) or tumor necrosis factor-α (TNF-α) in doses that increase mean blood pressure (MBP) and sympathetic drive increased the expression of SDF-1 in PVN. ICV administration of SDF-1 increased the phosphorylation of p44/42 mitogen-activated protein kinase (MAPK), JNK, and p38 MAPK in PVN, along with MBP, heart rate (HR), and renal sympathetic nerve activity (RSNA), but did not affect total p44/42 MAPK, JNK, and p38 MAPK levels. ICV pretreatment with the selective p44/42 MAPK inhibitor PD98059 prevented the SDF-1-induced increases in MBP, HR, and RSNA; ICV pretreatment with the selective JNK and p38 MAPK inhibitors attenuated but did not block these SDF-1-induced excitatory responses. ICV PD98059 also prevented the sympatho-excitatory response to bilateral PVN microinjections of SDF-1. ICV pretreatment with SDF-1 short-hairpin RNA significantly reduced ANG II- and TNF-α-induced phosphorylation of p44/42 MAPK in PVN. These findings identify TNF-α and ANG II as drivers of SDF-1 expression in PVN and suggest that the full expression of their cardiovascular and sympathetic effects depends upon SDF-1-mediated activation of p44/42 MAPK signaling.
Computational modeling of Takotsubo cardiomyopathy: effect of spatially varying β-adrenergic stimulation in the rat left ventricle
Authors: Land Sander , Niederer Steven A. , Louch William E. , Røe Åsmund T. , Aronsen Jan Magnus , Stuckey Daniel J. , Sikkel Markus B. , Tranter Matthew H. , Lyon Alexander R. , Harding Sian E. , Smith Nicolas P.
Published: September 2014

In Takotsubo cardiomyopathy, the left ventricle shows apical ballooning combined with basal hypercontractility. Both clinical observations in humans and recent experimental work on isolated rat ventricular myocytes suggest the dominant mechanisms of this syndrome are related to acute catecholamine overload. However, relating observed differences in single cells to the capacity of such alterations to result in the extreme changes in ventricular shape seen in Takotsubo syndrome is difficult. By using a computational model of the rat left ventricle, we investigate which mechanisms can give rise to the typical shape of the ventricle observed in this syndrome. Three potential dominant mechanisms related to effects of β-adrenergic stimulation were considered: apical-basal variation of calcium transients due to differences in L-type and sarco(endo)plasmic reticulum Ca2+-ATPase activation, apical-basal variation of calcium sensitivity due to differences in troponin I phosphorylation, and apical-basal variation in maximal active tension due to, e.g., the negative inotropic effects of p38 MAPK. Furthermore, we investigated the interaction of these spatial variations in the presence of a failing Frank-Starling mechanism. We conclude that a large portion of the apex needs to be affected by severe changes in calcium regulation or contractile function to result in apical ballooning, and smooth linear variation from apex to base is unlikely to result in the typical ventricular shape observed in this syndrome. A failing Frank-Starling mechanism significantly increases apical ballooning at end systole and may be an important additional factor underpinning Takotsubo syndrome.
Cardiac P2X purinergic receptors as a new pathway for increasing Na+ entry in cardiac myocytes
Authors: Shen Jian-Bing , Yang Ronghua , Pappano Achilles , Liang Bruce T.
Published: September 2014

P2X4 receptors (P2X4Rs) are ligand-gated ion channels capable of conducting cations such as Na+. Endogenous cardiac P2X4R can mediate ATP-activated current in adult murine cardiomyocytes. In the present study, we tested the hypothesis that cardiac P2X receptors can induce Na+ entry and modulate Na+ handling. We further determined whether P2X receptor-induced stimulation of the Na+/Ca2+ exchanger (NCX) has a role in modulating the cardiac contractile state. Changes in Na+-K+-ATPase current ( Ip) and NCX current ( INCX) after agonist stimulation were measured in ventricular myocytes of P2X4 transgenic mice using whole cell patch-clamp techniques. The agonist 2-methylthio-ATP (2-meSATP) increased peak Ip from a basal level of 0.52 ± 0.02 to 0.58 ± 0.03 pA/pF. 2-meSATP also increased the Ca2+ entry mode of INCX (0.55 ± 0.09 pA/pF under control conditions vs. 0.82 ± 0.14 pA/pF with 2-meSATP) at a membrane potential of +50 mV. 2-meSATP shifted the reversal potential of INCX from −14 ± 2.3 to −25 ± 4.1 mV, causing an estimated intracellular Na+ concentration increase of 1.28 ± 0.42 mM. These experimental results were closely mimicked by mathematical simulations based on previously established models. KB-R7943 or a structurally different agent preferentially opposing the Ca2+ entry mode of NCX, YM-244769, could inhibit the 2-meSATP-induced increase in cell shortening in transgenic myocytes. Thus, the Ca2+ entry mode of INCX participates in P2X agonist-stimulated contractions. In ventricular myocytes from wild-type mice, the P2X agonist could increase INCX, and KB-R7943 was able to inhibit the contractile effect of endogenous P2X4Rs, indicating a physiological role of these receptors in wild-type cells. The data demonstrate a novel Na+ entry pathway through ligand-gated P2X4Rs in cardiomyocytes.
Regulation of Notch 1 signaling in THP-1 cells enhances M2macrophage differentiation
Authors: Singla Reetu D. , Wang Jing , Singla Dinender K.
Published: September 2014

Macrophage polarization is emerging as an important area of research for the development of novel therapeutics to treat inflammatory diseases. Within the current study, the role of Notch1R in macrophage differentiation was investigated as well as downstream effects in THP-1 monocytes cultured in “inflammation mimicry” media. Interference of Notch signaling was achieved using either the pharmaceutical inhibitor DAPT or Notch1R small interfering RNA (siRNA), and Notch1R expression, macrophage phenotypes, and anti- and proinflammatory cytokine expression were evaluated. Data presented show that Notch1R expression on M1macrophages as well as M1macrophage differentiation is significantly elevated during cellular stress ( P < 0.05). However, under identical culture conditions, interference to Notch signaling via Notch1R inhibition mitigated these results as well as promoted M2macrophage differentiation. Moreover, when subjected to cellular stress, macrophage secretion of proinflammatory cytokines was significantly heightened ( P < 0.05). Importantly, Notch1R inhibition not only diminished proinflammatory cytokine secretion but also enhanced anti-inflammatory protein release ( P < 0.05). Our data suggest that Notch1R plays a pivotal role in M1macrophage differentiation and heightened inflammatory responses. Therefore, we conclude that inhibition of Notch1R and subsequent downstream signaling enhances monocyte to M2polarized macrophage outcomes and promotes anti-inflammatory mediation during cellular stress.
Granule cargo release from bone marrow-derived cells sustains cardiac hypertrophy
Authors: Yang Fanmuyi , Dong Anping , Ahamed Jasimuddin , Sunkara Manjula , Smyth Susan S.
Published: September 2014

Bone marrow-derived inflammatory cells, including platelets, may contribute to the progression of pressure overload-induced left ventricular hypertrophy (LVH). However, the underlying mechanisms for this are still unclear. One potential mechanism is through release of granule cargo. Unc13-d Jinx (Jinx) mice, which lack Munc13-4, a limiting factor in vesicular priming and fusion, have granule secretion defects in a variety of hematopoietic cells, including platelets. In the current study, we investigated the role of granule secretion in the development of LVH and cardiac remodeling using chimeric mice specifically lacking Munc13-4 in marrow-derived cells. Pressure overload was elicited by transverse aortic constriction (TAC). Chimeric mice were created by bone marrow transplantation. Echocardiography, histology staining, immunohistochemistry, real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and mass spectrometry were used to study LVH progression and inflammatory responses. Wild-type (WT) mice that were transplanted with WT bone marrow (WT→WT) and WT mice that received Jinx bone marrow (Jinx→WT) developed LVH and a classic fetal reprogramming response early (7 days) after TAC. However, at late times (5 wk), mice lacking Munc13-4 in bone marrow-derived cells (Jinx→WT) failed to sustain the cardiac hypertrophy observed in WT chimeric mice. No difference in cardiac fibrosis was observed at early or late time points. Reinjection of WT platelets or platelet releasate partially restored cardiac hypertrophy in Jinx chimeric mice. These results suggest that sustained LVH in the setting of pressure overload depends on one or more factors secreted from bone marrow-derived cells, possibly from platelets. Inhibiting granule cargo release may represent a novel target for preventing sustained LVH.
GSK3β-dependent inhibition of AMPK potentiates activation of neutrophils and macrophages and enhances severity of acute lung injury
Authors: Park Dae Won , Jiang Shaoning , Liu Yanping , Siegal Gene P. , Inoki Ken , Abraham Edward , Zmijewski Jaroslaw W.
Published: September 2014

Although AMP-activated protein kinase (AMPK) is involved in regulating carbohydrate and lipid metabolism, activated AMPK also plays an anti-inflammatory role in many cell populations. However, despite the ability of AMPK activation to diminish the severity of inflammatory responses, previous studies have found that AMPK activity is diminished in LPS-treated neutrophils and also in lungs of mice with LPS-induced acute lung injury (ALI). Since GSK3β participates in regulating AMPK activity, we examined potential roles for GSK3β in modulating LPS-induced activation of neutrophils and macrophages and in influencing severity of ALI. We found that GSK3β-dependent phosphorylation of T479-AMPK was associated with pT172 dephosphorylation and inactivation of AMPK following TLR4 engagement. GSK3β inhibitors BIO (6-bromoindirubin-3′-oxime), SB216763 , or siRNA knockdown of GSK3β, but not the PI3K/AKT inhibitor LY294002, prevented Thr172-AMPK dephosphorylation. Exposure to LPS resulted in rapid binding between IKKβ and AMPKα, and phosphorylation of S485-AMPK by IKKβ. These results suggest that IKKβ-dependent phosphorylation of S485-AMPK was an essential step in subsequent phosphorylation and inactivation AMPK by GSK3β. Inhibition of GSK3β activity delayed IκBα degradation and diminished expression of the proinflammatory TNF-α in LPS-stimulated neutrophils and macrophages. In vivo, inhibition of GSK3β decreased the severity of LPS-induced lung injury as assessed by development of pulmonary edema, production of TNF-α and MIP-2, and release of the alarmins HMGB1 and histone 3 in the lungs. These results show that inhibition of AMPK by GSK3β plays an important contributory role in enhancing LPS-induced inflammatory responses, including worsening the severity of ALI.
Impaired TLR4 and HIF expression in cystic fibrosis bronchial epithelial cells downregulates hemeoxygenase-1 and alters iron homeostasis in vitro
Authors: Chillappagari Shashi , Venkatesan Shalini , Garapati Virajith , Mahavadi Poornima , Munder Antje , Seubert Andreas , Sarode Gaurav , Guenther Andreas , Schmeck Bernd T. , Tümmler Burkhard , Henke Markus O.
Published: September 2014

Hemeoxygenase-1 (HO-1), an inducible heat shock protein, is upregulated in response to multiple cellular insults via oxidative stress, lipopolysaccharides (LPS), and hypoxia. In this study, we investigated in vitro the role of Toll-like receptor 4 (TLR4), hypoxia-inducible factor 1α (HIF-1α), and iron on HO-1 expression in cystic fibrosis (CF). Immunohistochemical analysis of TLR4, HO-1, ferritin, and HIF-1α were performed on lung sections of CFTR−/− and wild-type mice. CFBE41o- and 16HBE14o- cell lines were employed for in vitro analysis via immunoblotting, immunofluorescence, real-time PCR, luciferase reporter gene analysis, and iron quantification. We observed a reduced TLR4, HIF-1α, HO-1, and ferritin in CFBE41o- cell line and CF mice. Knockdown studies using TLR4-siRNA in 16HBE14o- revealed significant decrease of HO-1, confirming the role of TLR4 in HO-1 downregulation. Inhibition of HO-1 using tin protoporphyrin in 16HBE14o- cells resulted in increased iron levels, suggesting a probable role of HO-1 in iron accumulation. Additionally, sequestration of excess iron using iron chelators resulted in increased hypoxia response element response in CFBE41o- and 16HBE14o-, implicating a role of iron in HIF-1α stabilization and HO-1. To conclude, our in vitro results demonstrate that multiple regulatory factors, such as impaired TLR4 surface expression, increased intracellular iron, and decreased HIF-1α, downregulate HO-1 expression in CFBE41o- cells.
Effects of postexercise milk consumption on whole body protein balance in youth
Authors: Volterman Kimberly A. , Obeid Joyce , Wilk Boguslaw , Timmons Brian W.
Published: September 2014

In adults, adding protein to a postexercise beverage increases muscle protein turnover and replenishes amino acid stores. Recent focus has shifted toward the use of bovine-based milk and milk products as potential postexercise beverages; however, little is known about how this research translates to the pediatric population. Twenty-eight (15 girls) pre- to early pubertal (PEP, 7–11 yr) and mid- to late-pubertal (MLP, 14–17 yr) children consumed an oral dose of [15N]glycine prior to performing 2 × 20-min cycling bouts at 60% V̇o2 peakin a warm environment (34.5°C, 47.3% relative humidity). Following exercise, participants consumed either water (W), a carbohydrate-electrolyte solution (CES), or skim milk (SM) in a randomized, cross-over fashion in a volume equal to 100% of their body mass loss during exercise. Whole body nitrogen turnover (Q), protein synthesis (S), protein breakdown (B), and whole body protein balance (WBPB) were measured over 16 h. Protein intake from SM was 0.40 ± 0.10 g/kg. Over 16 h, Q and S were significantly greater ( P < 0.01) with SM than W and CES. B demonstrated a trend for a main effect for beverage ( P = 0.063). WBPB was more negative ( P < 0.01) with W and CES than with SM. In the SM trial, WBPB was positive in PEP, although it remained negative in MLP. Boys exhibited significantly more negative WBPB than girls ( P < 0.05). Postexercise milk consumption enhances WBPB compared with W and CES; however, additional protein intake may be required to sustain a net anabolic environment over 16 h.
BRCA1 Suppresses Epithelial-to-Mesenchymal Transition and Stem Cell Dedifferentiation during Mammary and Tumor Development
Authors: Bai Feng , Chan Ho Lam , Scott Alexandria , Smith Matthew D. , Fan Cheng , Herschkowitz Jason I. , Perou Charles M. , Livingstone Alan S. , Robbins David J. , Capobianco Anthony J. , Pei Xin-Hai
Published: September 2014

BRCA1 mutation carriers are predisposed to developing basal-like breast cancers with high metastasis and poor prognosis. Yet, how BRCA1 suppresses formation of basal-like breast cancers is still obscure. Deletion of p18Ink4c (p18), an inhibitor of CDK4 and CDK6, functionally inactivates the RB pathway, stimulates mammary luminal stem cell (LSC) proliferation, and leads to spontaneous luminal tumor development. Alternately, germline mutation of Brca1 shifts the fate of luminal cells to cause luminal-to-basal mammary tumor transformation. Here, we report that disrupting Brca1 by either germline or epithelium-specific mutation in p18-deficient mice activates epithelial-to-mesenchymal transition (EMT) and induces dedifferentiation of LSCs, which associate closely with expansion of basal and cancer stem cells and formation of basal-like tumors. Mechanistically, BRCA1 bound to the TWIST promoter, suppressing its activity and inhibiting EMT in mammary tumor cells. In human luminal cancer cells, BRCA1 silencing was sufficient to activate TWIST and EMT and increase tumor formation. In parallel, TWIST expression and EMT features correlated inversely with BRCA1 expression in human breast cancers. Together, our findings showed that BRCA1 suppressed TWIST and EMT, inhibited LSC dedifferentiation, and repressed expansion of basal stem cells and basal-like tumors. Thus, our work offers the first genetic evidence that Brca1 directly suppresses EMT and LSC dedifferentiation during breast tumorigenesis. Cancer Res; 74(21); 6161–72. ©2014 AACR.
Validation of Biomarkers That Complement CA19.9 in Detecting Early Pancreatic Cancer
Authors: Chan Alison , Prassas Ioannis , Dimitromanolakis Apostolos , Brand Randall E. , Serra Stefano , Diamandis Eleftherios P. , Blasutig Ivan M.
Published: September 2014

Purpose: Pancreatic ductal adenocarcinoma (PDAC) is a significant cause of cancer mortality. Carbohydrate antigen 19.9 (CA19.9), the only tumor marker available to detect and monitor PDAC, is not sufficiently sensitive and specific to consistently differentiate early cancer from benign disease. In this study, we aimed to validate recently discovered serum protein biomarkers for the early detection of PDAC and ultimately develop a biomarker panel that could discriminate PDAC from other benign disease better than the existing marker CA19.9.

Patients and Methods: We performed a retrospective blinded evaluation of 400 serum samples collected from individuals recruited on a consecutive basis. The sample population consisted of 250 individuals with PDAC at various stages, 130 individuals with benign conditions and 20 healthy individuals. The serum levels of each biomarker were determined by ELISAs or automated immunoassay.

Results: By randomly splitting matched samples into a training (n = 186) and validation (n = 214) set, we were able to develop and validate a biomarker panel consisting of CA19.9, CA125, and LAMC2 that significantly improved the performance of CA19.9 alone. Improved discrimination was observed in the validation set between all PDAC and benign conditions (AUCCA19.9 = 0.80 vs. AUCCA19.9+CA125+LAMC2 = 0.87; P < 0.005) as well as between early-stage PDAC and benign conditions (AUCCA19.9 = 0.69 vs. AUCCA19.9+CA125+LAMC2 = 0.76; P < 0.05) and between early-stage PDAC and chronic pancreatitis (CP; AUCCA19.9 = 0.59 vs. AUCCA19.9+CA125+LAMC2 = 0.74; P < 0.05).

Conclusions: The data demonstrate that a serum protein biomarker panel consisting of CA125, CA19.9, and LAMC2 is able to significantly improve upon the performance of CA19.9 alone in detecting PDAC. Clin Cancer Res; 20(22); 5787–95. ©2014 AACR.
Phase II Study of Perifosine and Sorafenib Dual-Targeted Therapy in Patients with Relapsed or Refractory Lymphoproliferative Diseases
Authors: Guidetti Anna , Carlo-Stella Carmelo , Locatelli Silvia L. , Malorni Walter , Mortarini Roberta , Viviani Simonetta , Russo Domenico , Marchianò Alfonso , Sorasio Roberto , Dodero Anna , Farina Lucia , Giordano Laura , Di Nicola Massimo , Anichini Andrea , Corradini Paolo , Gianni Alessandro M.
Published: September 2014

Purpose: To evaluate safety and activity of perifosine and sorafenib combination therapy in patients with lymphoproliferative diseases.

Experimental Design: Patients with relapsed and refractory lymphoproliferative diseases received perifosine (50 mg twice daily) for 1 month. Patients achieving less than partial response (PR) after perifosine alone were administered the combination therapy [perifosine plus sorafenib (400 mg twice daily)] until progressive disease (PD) or unacceptable toxicity occurred. The pERK and pAKT in peripheral blood lymphocytes as well as serum cytokine levels were investigated as predictive biomarkers of response.

Results: Forty patients enrolled in this study. After 1 month of perifosine alone, 36 who achieved less than PR went on to combination therapy, whereas four patients with chronic lymphocytic leukemia (CLL) who achieved PR continued with perifosine alone for a median of 10 months (range, 4–21). The most common drug-related toxicities were grade 1–2 anemia (17%), thrombocytopenia (9%), diarrhea (25%), joint pain (22%), and hand–foot skin reaction (25%). Three patients experienced grade 3 pneumonitis. Eight patients (22%) achieved PR, 15 (42%) achieved stable disease, and 13 (36%) experienced PD. A 28% PR rate was recorded for 25 patients with Hodgkin lymphoma. Among all patients, median overall survival and progression-free survival were 16 and 5 months, respectively. Early reductions in pERK and pAKT significantly correlated with the probability of clinical response.

Conclusions: Perifosine and sorafenib combination therapy is feasible with manageable toxicity and demonstrates promising activity in patients with Hodgkin lymphoma. The predictive value of pERK and pAKT should be confirmed in a larger patient cohort. Clin Cancer Res; 20(22); 5641–51. ©2014 AACR.
Interrogating Two Schedules of the AKT Inhibitor MK-2206 in Patients with Advanced Solid Tumors Incorporating Novel Pharmacodynamic and Functional Imaging Biomarkers
Authors: Yap Timothy A. , Yan Li , Patnaik Amita , Tunariu Nina , Biondo Andrea , Fearen Ivy , Papadopoulos Kyriakos P. , Olmos David , Baird Richard , Delgado Liliana , Tetteh Ernestina , Beckman Robert A. , Lupinacci Lisa , Riisnaes Ruth , Decordova Shaun , Heaton Simon P. , Swales Karen , deSouza Nandita M. , Leach Martin O. , Garrett Michelle D. , Sullivan Daniel M. , de Bono Johann S. , Tolcher Anthony W.
Published: September 2014

Purpose: Multiple cancers harbor genetic aberrations that impact AKT signaling. MK-2206 is a potent pan-AKT inhibitor with a maximum tolerated dose (MTD) previously established at 60 mg on alternate days (QOD). Due to a long half-life (60–80 hours), a weekly (QW) MK-2206 schedule was pursued to compare intermittent QW and continuous QOD dosing.

Experimental Design: Patients with advanced cancers were enrolled in a QW dose-escalation phase I study to investigate the safety and pharmacokinetic–pharmacodynamic profiles of tumor and platelet-rich plasma (PRP). The QOD MTD of MK-2206 was also assessed in patients with ovarian and castration-resistant prostate cancers and patients with advanced cancers undergoing multiparametric functional magnetic resonance imaging (MRI) studies, including dynamic contrast-enhanced MRI, diffusion-weighted imaging, magnetic resonance spectroscopy, and intrinsic susceptibility-weighted MRI.

Results: A total of 71 patients were enrolled; 38 patients had 60 mg MK-2206 QOD, whereas 33 received MK-2206 at 90, 135, 150, 200, 250, and 300 mg QW. The QW MK-2206 MTD was established at 200 mg following dose-limiting rash at 250 and 300 mg. QW dosing appeared to be similarly tolerated to QOD, with toxicities including rash, gastrointestinal symptoms, fatigue, and hyperglycemia. Significant AKT pathway blockade was observed with both continuous QOD and intermittent QW dosing of MK-2206 in serially obtained tumor and PRP specimens. The functional imaging studies demonstrated that complex multiparametric MRI protocols may be effectively implemented in a phase I trial.

Conclusions: Treatment with MK-2206 safely results in significant AKT pathway blockade in QOD and QW schedules. The intermittent dose of 200 mg QW is currently used in phase II MK-2206 monotherapy and combination studies (NCT00670488). Clin Cancer Res; 20(22); 5672–85. ©2014 AACR.
Ponatinib Inhibits Polyclonal Drug-Resistant KIT Oncoproteins and Shows Therapeutic Potential in Heavily Pretreated Gastrointestinal Stromal Tumor (GIST) Patients
Authors: Garner Andrew P. , Gozgit Joseph M. , Anjum Rana , Vodala Sadanand , Schrock Alexa , Zhou Tianjun , Serrano Cesar , Eilers Grant , Zhu Meijun , Ketzer Julia , Wardwell Scott , Ning Yaoyu , Song Youngchul , Kohlmann Anna , Wang Frank , Clackson Tim , Heinrich Michael C. , Fletcher Jonathan A. , Bauer Sebastian , Rivera Victor M.
Published: September 2014

Purpose: KIT is the major oncogenic driver of gastrointestinal stromal tumors (GIST). Imatinib, sunitinib, and regorafenib are approved therapies; however, efficacy is often limited by the acquisition of polyclonal secondary resistance mutations in KIT, with those located in the activation (A) loop (exons 17/18) being particularly problematic. Here, we explore the KIT-inhibitory activity of ponatinib in preclinical models and describe initial characterization of its activity in patients with GIST.

Experimental Design: The cellular and in vivo activities of ponatinib, imatinib, sunitinib, and regorafenib against mutant KIT were evaluated using an accelerated mutagenesis assay and a panel of engineered and GIST-derived cell lines. The ponatinib–KIT costructure was also determined. The clinical activity of ponatinib was examined in three patients with GIST previously treated with all three FDA-approved agents.

Results: In engineered and GIST-derived cell lines, ponatinib potently inhibited KIT exon 11 primary mutants and a range of secondary mutants, including those within the A-loop. Ponatinib also induced regression in engineered and GIST-derived tumor models containing these secondary mutations. In a mutagenesis screen, 40 nmol/L ponatinib was sufficient to suppress outgrowth of all secondary mutants except V654A, which was suppressed at 80 nmol/L. This inhibitory profile could be rationalized on the basis of structural analyses. Ponatinib (30 mg daily) displayed encouraging clinical activity in two of three patients with GIST.

Conclusion:Ponatinib possesses potent activity against most major clinically relevant KIT mutants and has demonstrated preliminary evidence of activity in patients with refractory GIST. These data strongly support further evaluation of ponatinib in patients with GIST. Clin Cancer Res; 20(22); 5745–55. ©2014 AACR.
In Vitro and In Vivo Interactions between the HDAC6 Inhibitor Ricolinostat (ACY1215) and the Irreversible Proteasome Inhibitor Carfilzomib in Non-Hodgkin Lymphoma Cells
Authors: Dasmahapatra Girija , Patel Hiral , Friedberg Johnathan , Quayle Steven N. , Jones Simon S. , Grant Steven
Published: September 2014

Interactions between the HDAC6 inhibitor ricolinostat (ACY1215) and the irreversible proteasome inhibitor carfilzomib were examined in non-Hodgkin lymphoma (NHL) models, including diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), and double-hit lymphoma cells. Marked in vitro synergism was observed in multiple cell types associated with activation of cellular stress pathways (e.g., JNK1/2, ERK1/2, and p38) accompanied by increases in DNA damage (γH2A.X), G2–M arrest, and the pronounced induction of mitochondrial injury and apoptosis. Combination treatment with carfilzomib and ricolinostat increased reactive oxygen species (ROS), whereas the antioxidant TBAP attenuated DNA damage, JNK activation, and cell death. Similar interactions occurred in bortezomib-resistant and double-hit DLBCL, MCL, and primary DLBCL cells, but not in normal CD34+ cells. However, ricolinostat did not potentiate inhibition of chymotryptic activity by carfilzomib. shRNA knockdown of JNK1 (but not MEK1/2), or pharmacologic inhibition of p38, significantly reduced carfilzomib–ricolinostat lethality, indicating a functional contribution of these stress pathways to apoptosis. Combined exposure to carfilzomib and ricolinostat also markedly downregulated the cargo-loading protein HR23B. Moreover, HR23B knockdown significantly increased carfilzomib- and ricolinostat-mediated lethality, suggesting a role for this event in cell death. Finally, combined in vivo treatment with carfilzomib and ricolinostat was well tolerated and significantly suppressed tumor growth and increased survival in an MCL xenograft model. Collectively, these findings indicate that carfilzomib and ricolinostat interact synergistically in NHL cells through multiple stress-related mechanisms, and suggest that this strategy warrants further consideration in NHL. Mol Cancer Ther; 13(12); 2886–97. ©2014 AACR.
Development of Human Serine Protease-Based Therapeutics Targeting Fn14 and Identification of Fn14 as a New Target Overexpressed in TNBC
Authors: Zhou Hong , Mohamedali Khalid A. , Gonzalez-Angulo Ana Maria , Cao Yu , Migliorini Mary , Cheung Lawrence H. , LoBello Janine , Lei Xiudong , Qi Yuan , Hittelman Walter N. , Winkles Jeffrey A. , Tran Nhan L. , Rosenblum Michael G.
Published: September 2014

The cytokine TWEAK and its receptor, Fn14, have emerged as potentially valuable targets for cancer therapy. Granzyme B (GrB)–containing Fn14-targeted constructs were generated containing either the Fn14 ligand TWEAK (GrB-TWEAK) or an anti-Fn14 humanized single-chain antibody (GrB-Fc-IT4) as the targeting moieties. Both constructs showed high affinity and selective cytotoxicity against a panel of Fn14-expressing human tumor cells including triple-negative breast cancer (TNBC) lines. Cellular expression of the GrB inhibitor PI-9 in target cells had no impact on the cytotoxic effect of either construct. Cellular expression of MDR1 showed no cross-resistance to the fusion constructs. GrB-TWEAK and GrB-Fc-IT4 activated intracellular caspase cascades and cytochrome c–related proapoptotic pathways consistent with the known intracellular functions of GrB in target cells. Treatment of mice bearing established HT-29 xenografts with GrB-TWEAK showed significant tumor growth inhibition compared with vehicle alone (P < 0.05). Both GrB-TWEAK and GrB-Fc-IT4 displayed significant tumor growth inhibition when administered to mice bearing orthotopic MDA-MB-231 (TNBC) tumor xenografts. The Cancer Genome Atlas analysis revealed that Fn14 mRNA expression was significantly higher in TNBC and in HER2-positive disease (P < 0.0001) compared with hormone receptor–positive breast cancer, and in basal-like 2 tumors (P = 0.01) compared with other TNBC molecular subtypes. IHC analysis of a 101 patient TNBC tumor microarray showed that 55 of 101 (54%) of tumors stained positive for Fn14, suggesting that this may be an excellent potential target for precision therapeutic approaches. Targeting Fn14 using fully human, GrB-containing fusion constructs may form the basis for a new class of novel, potent, and highly effective constructs for targeted therapeutic applications. Mol Cancer Ther; 13(11); 2688–705. ©2014 AACR.
Development of Targeted Near-Infrared Imaging Agents for Prostate Cancer
Authors: Wang Xinning , Huang Steve S. , Heston Warren D.W. , Guo Hong , Wang Bing-Cheng , Basilion James P.
Published: September 2014

Prostate cancer is the most common noncutaneous malignancy affecting men in North America. Radical prostatectomy remains a definitive treatment for prostate cancer. However, prostate surgeries are still performed “blindly” with the extent of tumor infiltration past the margins of the surgery only being determined postoperatively. An imaging modality that can be used during surgery is needed to help define the tumor margins. With its abundant expression in prostate cancer, prostate-specific membrane antigen (PSMA) is an ideal target for detection of prostate cancer. The purpose of this study was to develop PSMA-targeted near-infrared (NIR) optical imaging probes for intraoperative visualization of prostate cancer. We synthesized a high-affinity PSMA ligand (PSMA-1) with low molecular weight and further labeled it with commercially available NIR dyes IRDy800 and Cy5.5. PSMA-1 and PSMA-1–NIR conjugates had binding affinities better than the parent ligand Cys-CO-Glu. Selective binding was measured for each of the probes in both in vitro and in vivo studies using competitive binding and uptake studies. Interestingly, the results indicated that the pharmacokinetics of the probes was dependent of the fluorophore conjugated to the PSMA-1 ligand and varied widely. These data suggest that PSMA-targeted probes have the potential to be further developed as contrast agents for clinical intraoperative fluorescence-guided surgery. Mol Cancer Ther; 13(11); 2595–606. ©2014 AACR.
Cancer-Related Therapies at the End of Life in Hospitalized Cancer Patients from Four Swiss Cantons: SAKK 89/09
Authors: Matter-Walstra Klazien W. , Achermann Rita , Rapold Roland , Klingbiel Dirk , Bordoni Andrea , Dehler Silvia , Jundt Gernot , Konzelmann Isabelle , Clough-Gorr Kerri , Szucs Thomas , Pestalozzi Bernhard C. , Schwenkglenks Matthias
Published: September 2014

The use of cancer-related therapies in cancer patients hospitalized at the end of life has increased in many countries over time. Given the scarcity of published Swiss data, the objective of this study was to evaluate the influence of hospital type and other factors on the delivery of health care during the last month before death. Claims data were used to assess health care utilization of cancer patients (identified by cancer registry data of four participating Swiss cantons) who deceased between 2006 and 2008. Primary endpoints were delivery of cancer-related therapies during the last 30 days before death. Multivariate logistic regression assessed the explanatory value of hospital type, patient and geographic characteristics. Of 3,809 identified cancer patients in the claims database, 2,086 patients dying from cancer were hospitalized during the last 30 days before death, generating 2,262 inpatient episodes. Anticancer drug therapy was given in 22.2% and radiotherapy in 11.7% of episodes. Besides age and cancer type, the canton of residence and hospital type showed independent, statistically significant associations with intensity of care, which was highest in university hospitals. These results should initiate a discussion among oncologists in Switzerland and may question the compliance with standard of care guidelines for terminal cancer patients. i 2014 S. Karger AG, Basel
Activities of the European Multiple Sclerosis Platform
Authors: Rogan Emma
Published: September 2014

Background: The European Multiple Sclerosis Platform (EMSP) is an umbrella organization for national multiple sclerosis (MS) patients' associations in 34 European countries. Its focus is to improve the quality of life (QoL) of individuals with MS and inform society about the challenges of living with this chronic and debilitating disease. Summary: The EMSP, which is comprised of 39 MS societies across Europe, represents the interests of more than 600,000 individuals with MS and another 1.5 million persons affected by the disease, including family and caregivers. The organization works to achieve high quality, equitable treatment and support for persons with MS throughout Europe, as reflected in its main activities and projects. These include: the European Code of Good Practice in MS; a European MS register (EUReMS); a MS Barometer to assess QoL; the MS Nurse PROfessional curriculum to provide standards for MS nurse training; a photographic exhibition entitled Under Pressure; the Believe and Achieve project which focuses on employment for young people with MS; and collaboration in World MS Day, an international MS awareness day. Key Messages: The activities and projects of the EMSP are geared towards making a difference in the lives of individuals with MS in Europe and their wider support networks, including physicians, neurologists, nurses and carers.
Variations in the Blood Phenytoin Levels during Long-Term Combined Treatment with S-1 and Phenytoin
Authors: Negoro Yutaka , Higashi Takashi , Matsuoka Hiroaki , Watanabe Kyohei , Igarashi Toshiaki , Kayano Yuichiro , Yano Ryoichi , Nakamura Toshiaki , Masada Mikio
Published: September 2014

Although combination therapy with the oral fluoropyrimidine anticancer drug S-1 and the anticonvulsant phenytoin (PHT) is known to increase blood levels of PHT and the risk of intoxication, reports on long-term monitoring of blood levels of PHT during combined S-1 and PHT treatment and a thorough understanding of their interaction are lacking. This report aims to describe interactive effects of S-1 and PHT through long-term therapeutic drug monitoring of PHT. A 72-year-old male had been prescribed oral PHT (130 mg/day) for over 20 years and started receiving S-1 therapy (80 mg/day for 4 weeks, followed by a 2-week rest) as postoperative adjuvant chemotherapy for gastric cancer. The blood PHT level was continuously monitored. Prior to receiving S-1, the patient's blood PHT concentration was 6.0 μg/ml, but it increased during S-1 therapy, reaching 22.9 μg/ml on day 84 (during a rest period of second cycle S-1 therapy). After reducing his PHT dosage to 100 mg/day, it never reached toxic levels (4.0-10.4 μg/ml). It was difficult to keep blood PHT concentrations constant because of the time lag between the period of combined use of S-1 and PHT and the timing of manifestation and disappearance of the drug interaction. The DIPS probability scale indicated a highly probable interaction between S-1 and PHT. We conclude that, when S-1 and PHT are used concurrently, occurrence and disappearance time of their interaction need to be predicted to maintain an effective and safe PHT concentration.
Reversal of Hypoxia in Murine Atherosclerosis Prevents Necrotic Core Expansion by Enhancing Efferocytosis
Authors: Marsch Elke , Theelen Thomas L. , Demandt Jasper A.F. , Jeurissen Mike , van Gink Mathijs , Verjans Robin , Janssen Anique , Cleutjens Jack P. , Meex Steven J.R. , Donners Marjo M. , Haenen Guido R. , Schalkwijk Casper G. , Dubois Ludwig J. , Lambin Philippe , Mallat Ziad , Gijbels Marion J. , Heemskerk Johan W.M. , Fisher Edward A. , Biessen Erik A.L. , Janssen Ben J. , Daemen Mat J.A.P. , Sluimer Judith C.
Published: September 2014

Objective—

Advanced murine and human plaques are hypoxic, but it remains unclear whether plaque hypoxia is causally related to atherogenesis. Here, we test the hypothesis that reversal of hypoxia in atherosclerotic plaques by breathing hyperoxic carbogen gas will prevent atherosclerosis.

Approach and Results—

Low-density lipoprotein receptor–deficient mice (LDLR
-/-
) were fed a Western-type diet, exposed to carbogen (95% O
2
, 5% CO
2
) or air, and the effect on plaque hypoxia, size, and phenotype was studied. First, the hypoxic marker pimonidazole was detected in murine LDLR
-/-
plaque macrophages from plaque initiation onwards. Second, the efficacy of breathing carbogen (90 minutes, single exposure) was studied. Compared with air, carbogen increased arterial blood pO
2
5-fold in LDLR
-/-
mice and reduced plaque hypoxia in advanced plaques of the aortic root (−32%) and arch (−84%). Finally, the effect of repeated carbogen exposure on progression of atherosclerosis was studied in LDLR
-/-
mice fed a Western-type diet for an initial 4 weeks, followed by 4 weeks of diet and carbogen or air (both 90 min/d). Carbogen reduced plaque hypoxia (−40%), necrotic core size (−37%), and TUNEL
+
(terminal uridine nick-end labeling positive) apoptotic cell content (−50%) and increased efferocytosis of apoptotic cells by cluster of differentiation 107b
+
(CD107b, MAC3) macrophages (+36%) in advanced plaques of the aortic root. Plaque size, plasma cholesterol, hematopoiesis, and systemic inflammation were unchanged. In vitro, hypoxia hampered efferocytosis by bone marrow–derived macrophages, which was dependent on the receptor Mer tyrosine kinase.

Conclusions—

Carbogen restored murine plaque oxygenation and prevented necrotic core expansion by enhancing efferocytosis, likely via Mer tyrosine kinase. Thus, plaque hypoxia is causally related to necrotic core expansion.