bluuebunny/crossref_metadata_2025 huggingface dataset:
Oncogenic KRAS regulates BMP4 expression in colon cancer cell lines
Authors: Duerr Eva-Maria , Mizukami Yusuke , Moriichi Kentaro , Gala Manish , Jo Won-Seok , Kikuchi Hirotoshi , Xavier Ramnik J. , Chung Daniel C.
Published: March 2012

Activating mutations in the KRAS oncogene are common in colorectal cancer. However, the complete spectrum of KRAS targets that mediate its tumorigenic effect has not yet been fully delineated. We identified bone morphogenetic protein 4 (Bmp4), a transforming growth factor-β family member that regulates development and tissue homeostasis, as a new target of KRAS. In SW480, Hela, and 293 cells, oncogenic KRASV12downregulated BMP4 RNA levels, a BMP4 promoter luciferase construct, and Bmp4 protein levels. The MEK inhibitor PD98059 but not the phosphatidylinositol 3-kinase inhibitor LY294002 blocked this downregulation of BMP4. To identify the region of the BMP4 promoter that mediated this regulation by KRAS, serial 5′-deletions of the promoter were generated. An inhibitory region was identified between −3,285 and −3,258 bp in the Bmp4 promoter. In summary, oncogenic KRAS can downregulate Bmp4 through a transcriptional pathway that depends on ERK. These findings point to a unique link between two pathways that are frequently altered in colon cancer.
P2X7 receptor-mediated purinergic signaling promotes liver injury in acetaminophen hepatotoxicity in mice
Authors: Hoque Rafaz , Sohail Muhammed Adnan , Salhanick Steven , Malik Ahsan F. , Ghani Ayaz , Robson Simon C. , Mehal Wajahat Z.
Published: March 2012

Inflammation contributes to liver injury in acetaminophen (APAP) hepatotoxicity in mice and is triggered by stimulation of immune cells. The purinergic receptor P2X7 is upstream of the nod-like receptor family, pryin domain containing-3 (NLRP3) inflammasome in immune cells and is activated by ATP and NAD that serve as damage-associated molecular patterns. APAP hepatotoxicity was assessed in mice genetically deficient in P2X7, the key inflammatory receptor for nucleotides ( P2X7−/−), and in wild-type mice. P2X7−/− mice had significantly decreased APAP-induced liver necrosis. In addition, APAP-poisoned mice were treated with the specific P2X7 antagonist A438079 or etheno-NAD, a competitive antagonist of NAD. Pre- or posttreatment with A438079 significantly decreased APAP-induced necrosis and hemorrhage in APAP liver injury in wild-type but not P2X7−/− mice. Pretreatment with etheno-NAD also significantly decreased APAP-induced necrosis and hemorrhage in APAP liver injury. In addition, APAP toxicity in mice lacking the plasma membrane ecto-NTPDase CD39 ( CD39−/−) that metabolizes ATP was examined in parallel with the use of soluble apyrase to deplete extracellular ATP in wild-type mice. CD39−/− mice had increased APAP-induced hemorrhage and mortality, whereas apyrase also decreased APAP-induced mortality. Kupffer cells were treated with extracellular ATP to assess P2X7-dependent inflammasome activation. P2X7 was required for ATP-stimulated IL-1β release. In conclusion, P2X7 and exposure to the ligands ATP and NAD are required for manifestations of APAP-induced hepatotoxicity.
Coordination of motilin and ghrelin regulates the migrating motor complex of gastrointestinal motility in Suncus murinus
Authors: Mondal Anupom , Xie Zuoyun , Miyano Yuki , Tsutsui Chihiro , Sakata Ichiro , Kawamoto Yoichi , Aizawa Sayaka , Tanaka Toru , Oda Sen-ichi , Sakai Takafumi
Published: March 2012

Motilin and ghrelin are the gastrointestinal (GI) hormones released in a fasting state to stimulate the GI motility of the migrating motor complex (MMC). We focused on coordination of the ghrelin/motilin family in gastric contraction in vivo and in vitro using the house musk shrew ( Suncus murinus ), a ghrelin- and motilin-producing mammal. To measure the contractile activity of the stomach in vivo, we recorded GI contractions either in the free-moving conscious or anesthetized S. murinus and examined the effects of administration of motilin and/or ghrelin on spontaneous MMC in the fasting state. In the in vitro study, we also studied the coordinative effect of these hormones on the isolated stomach using an organ bath. In the fasting state, phase I, II, and III contractions were clearly recorded in the gastric body (as observed in humans and dogs). Intravenous infusion of ghrelin stimulated gastric contraction in the latter half of phase I and in the phase II in a dose-dependent manner. Continuous intravenous infusion of ghrelin antagonist (d-Lys3-GHRP6) significantly suppressed spontaneous phase II contractions and prolonged the time of occurrence of the peak of phase III contractions. However, intravenous infusion of motilin antagonist (MA-2029) did not inhibit phase II contractions but delayed the occurrence of phase III contractions of the MMC. In the in vitro study, even though a high dose of ghrelin did not stimulate contraction of stomach preparations, ghrelin administration (10−10-10−7M) with pretreatment of a low dose of motilin (10−10M) induced gastric contraction in a dose-dependent manner. Pretreatment with 10−8M ghrelin enhanced motilin-stimulated gastric contractions by 10 times. The interrelation of these peptides was also demonstrated in the anesthetized S. murinus . The results suggest that ghrelin is important for the phase II contraction and that coordination of motilin and ghrelin are necessary to initiate phase III contraction of the MMC.
p38 MAPK regulates Bax activity and apoptosis in enterocytes at baseline and after intestinal resection
Authors: Wakeman Derek , Guo Jun , Santos Jethrina A. , Wandu Wambui S. , Schneider John E. , McMellen Mark E. , Leinicke Jennifer A. , Erwin Christopher R. , Warner Brad W.
Published: March 2012

Increased apoptosis in crypt enterocytes is a key feature of intestinal adaptation following massive small bowel resection (SBR). Expression of the proapoptotic factor Bax has been shown to be required for resection-induced apoptosis. It has also been demonstrated that p38-α MAPK (p38) is necessary for Bax activation and apoptosis in vitro. The present studies were designed to test the hypothesis that p38 is a key regulator of Bax activation during adaptation after SBR in vivo. Enterocyte expression of p38 was deleted by tamoxifen administration to activate villin-Cre in adult mice with a floxed Mapk14 (p38-α) gene. Proximal 50% SBR or sham operations were performed on wild-type (WT) and p38 intestinal knockout (p38-IKO) mice under isoflurane anesthesia. Mice were killed 3 or 7 days after operation, and adaptation was analyzed by measuring intestinal morphology, proliferation, and apoptosis. Bax activity was quantified by immunoprecipitation, followed by Western blotting. After SBR, p38-IKO mice had deeper crypts, longer villi, and accelerated proliferation compared with WT controls. Rates of crypt apoptosis were significantly lower in p38-IKO mice, both at baseline and after SBR. Levels of activated Bax were twofold higher in WT mice after SBR relative to sham. In contrast, activated Bax levels were reduced by 67% in mice after p38 MAPK deletion. Deleted p38 expression within the intestinal epithelium leads to enhanced adaptation and reduced levels of enterocyte apoptosis after massive intestinal resection. p38-regulated Bax activation appears to be an important mechanism underlying resection-induced apoptosis.
DMT1 (IRE) expression in intestinal and erythroid cells is regulated by peripheral benzodiazepine receptor-associated protein 7
Authors: Okazaki Yasumasa , Ma Yuxiang , Yeh Mary , Yin Hong , Li Zhen , Yeh Kwo-yih , Glass Jonathan
Published: March 2012

The divalent metal transporter 1 (DMT1) is essential for cellular uptake of iron, mediating iron absorption across the duodenal brush border membrane. We have previously shown that with iron feeding DMT1 in the brush border membrane undergoes endocytosis into the subapical compartment of enterocytes. To understand the mechanisms of iron-induced endocytosis of DMT1, we used the yeast two-hybrid system to find proteins that interact with DMT1 and isolated from a rat duodenal cDNA library a protein that interacts specifically with the IRE containing isoform of DMT1 {DMT1 [iron-responsive element (IRE)]}. The protein (Genbank AY336075 ) is 97.5% identical with peripheral benzodiazepine receptor-associated protein 7 (PAP7), a protein that interacts with the peripheral benzodiazepine receptor. PAP7 is ubiquitously expressed in the rat and in multiple cell lines with consensus sequences including a nuclear localization signal and a Golgi dynamic domain. PAP7, expressed on the brush border of rat duodenum, copurified with DMT1 in brush border membrane vesicles, and following iron feeding, was internalized in parallel with the internalization of DMT1. To determine if PAP7 plays a role in cellular iron metabolism, we downregulated PAP7 expression in K562 cells with small interfering RNA. Following the decrease in PAP7 protein, DMT1 (IRE) protein but not mRNA was significantly downregulated but without effect on DMT1 (non-IRE), transferin (Tf)R1, or ferritin expression. Lowered levels of PAP7 resulted also in decreased cell proliferation and G1cell cycle arrest. These data are consistent with PAP7 interacting with DMT1 (IRE) and regulating DMT1 (IRE) expression in K562 cells by modulating expression of DMT1 (IRE) protein.
Oral vitamin C enhances the adrenergic vasoconstrictor response to local cooling in human skin
Authors: Yamazaki Fumio
Published: March 2012

Local administration of ascorbic acid (Asc) at a supraphysiological concentration inhibits the cutaneous vasoconstrictor response to local cooling (LC). However, whether orally ingesting Asc inhibits the LC-induced vasoconstrictor response remains unknown. The purpose of the present study was to examine the acute influence of oral Asc on the adrenergic vasoconstrictor response to LC in human skin. In experiment 1, skin blood flow (SkBF) was measured by laser-Doppler flowmetry at three sites (forearm, calf, palm). The three skin sites were locally cooled from 34 to 24°C at −1°C/min and maintained at 24°C for 20 min before (Pre) and 1.5 h after (Post) oral Asc (2-g single dose) or placebo supplementation. Cutaneous vascular conductance (CVC) was calculated as the ratio of SkBF to blood pressure and expressed relative to the baseline value before LC. Oral Asc enhanced ( P < 0.05) the reductions in CVC in the forearm (Pre, −50.3 ± 3.3%; Post, −57.8 ± 2.2%), calf (Pre, −52.6 ± 3.7%; Post, −66.1 ± 4.3%), and palm (Pre, −46.2 ± 6.2%; Post, −60.4 ± 5.6%) during LC. The placebo did not change the responses at any site. In experiment 2, to examine whether the increased vasoconstrictor response caused by oral Asc is due to the adrenergic system, the release of neurotransmitters from adrenergic nerves in forearm skin was blocked locally by iontophoresis of bretylium tosylate (BT). Oral Asc enhanced ( P < 0.05) the reductions in CVC at untreated control sites but did not change the responses at BT-treated sites during LC. In experiment 3, to further examine whether adrenergically mediated vasoconstriction is enhanced by oral Asc, 0.1 mM tyramine was administered using intradermal microdialysis in the forearm skin at 34°C in the Pre and Post periods. Oral Asc increased ( P < 0.05) the tyramine-induced reduction in CVC. These findings suggest that oral Asc acutely enhances the cutaneous vasoconstrictor responses to LC through the modification of adrenergic sympathetic mechanisms.
Unloading stress disturbs muscle regeneration through perturbed recruitment and function of macrophages
Authors: Kohno Shohei , Yamashita Yui , Abe Tomoki , Hirasaka Katsuya , Oarada Motoko , Ohno Ayako , Teshima-Kondo Shigetada , Higashibata Akira , Choi Inho , Mills Edward M. , Okumura Yuushi , Terao Junji , Nikawa Takeshi
Published: March 2012

Skeletal muscle is one of the most sensitive tissues to mechanical loading, and unloading inhibits the regeneration potential of skeletal muscle after injury. This study was designed to elucidate the specific effects of unloading stress on the function of immunocytes during muscle regeneration after injury. We examined immunocyte infiltration and muscle regeneration in cardiotoxin (CTX)-injected soleus muscles of tail-suspended (TS) mice. In CTX-injected TS mice, the cross-sectional area of regenerating myofibers was smaller than that of weight-bearing (WB) mice, indicating that unloading delays muscle regeneration following CTX-induced skeletal muscle damage. Delayed infiltration of macrophages into the injured skeletal muscle was observed in CTX-injected TS mice. Neutrophils and macrophages in CTX-injected TS muscle were presented over a longer period at the injury sites compared with those in CTX-injected WB muscle. Disturbance of activation and differentiation of satellite cells was also observed in CTX-injected TS mice. Further analysis showed that the macrophages in soleus muscles were mainly Ly-6C-positive proinflammatory macrophages, with high expression of tumor necrosis factor-α and interleukin-1β, indicating that unloading causes preferential accumulation and persistence of proinflammatory macrophages in the injured muscle. The phagocytic and myotube formation properties of macrophages from CTX-injected TS skeletal muscle were suppressed compared with those from CTX-injected WB skeletal muscle. We concluded that the disturbed muscle regeneration under unloading is due to impaired macrophage function, inhibition of satellite cell activation, and their cooperation.
Sex-based comparisons of myofibrillar protein synthesis after resistance exercise in the fed state
Authors: West Daniel W. D. , Burd Nicholas A. , Churchward-Venne Tyler A. , Camera Donny M. , Mitchell Cameron J. , Baker Steven K. , Hawley John A. , Coffey Vernon G. , Phillips Stuart M.
Published: March 2012

We made sex-based comparisons of rates of myofibrillar protein synthesis (MPS) and anabolic signaling after a single bout of high-intensity resistance exercise. Eight men (20 ± 10 yr, BMI = 24.3 ± 2.4) and eight women (22 ± 1.8 yr, BMI = 23.0 ± 1.9) underwent primed constant infusions of l-[ ring-13C6]phenylalanine on consecutive days with serial muscle biopsies. Biopsies were taken from the vastus lateralis at rest and 1, 3, 5, 24, 26, and 28 h after exercise. Twenty-five grams of whey protein was ingested immediately and 26 h after exercise. We also measured exercise-induced serum testosterone because it is purported to contribute to increases in myofibrillar protein synthesis (MPS) postexercise and its absence has been hypothesized to attenuate adaptative responses to resistance exercise in women. The exercise-induced area under the testosterone curve was 45-fold greater in men than women in the early (1 h) recovery period following exercise ( P < 0.001). MPS was elevated similarly in men and women (2.3- and 2.7-fold, respectively) 1–5 h postexercise and after protein ingestion following 24 h recovery. Phosphorylation of mTORSer2448 was elevated to a greater extent in men than women acutely after exercise ( P = 0.003), whereas increased phosphorylation of p70S6K1Thr389 was not different between sexes. Androgen receptor content was greater in men (main effect for sex, P = 0.049). Atrogin-1 mRNA abundance was decreased after 5 h recovery in both men and women ( P < 0.001), and MuRF-1 expression was elevated in men after protein ingestion following 24 h recovery ( P = 0.003). These results demonstrate minor sex-based differences in signaling responses and no difference in the MPS response to resistance exercise in the fed state. Interestingly, our data demonstrate that exercise-induced increases in MPS are dissociated from postexercise testosteronemia and that stimulation of MPS occurs effectively with low systemic testosterone concentrations in women.
Airway responsiveness depends on the diffusion rate of methacholine across the airway wall
Authors: Bates Jason H. T. , Stevenson Chelsea A. , Aliyeva Minara , Lundblad Lennart K. A.
Published: March 2012

During methacholine challenge tests of airway responsiveness, it is invariably assumed that the administered dose of agonist is accurately reflected in the dose that eventually reaches the airway smooth muscle (ASM). However, agonist must traverse a variety of tissue obstacles to reach the ASM, during which the agonist is subjected to both enzymatic breakdown and removal by the bronchial and pulmonary circulations. This raises the possibility that a significant fraction of the deposited agonist may never actually make it to the ASM. To understand the nature of this effect, we measured the time course of changes in airway resistance elicited by various durations of methacholine aerosol in mice. We fit to these data a computational model of a dynamically contracting airway responding to agonist that diffuses through an airway compartment, thereby obtaining rate constants that reflect the diffusive barrier to methacholine. We found that these barriers can contribute significantly to the time course of airway narrowing, raising the important possibility that alterations in the diffusive barrier presented by the airway wall may play a role in pathologically altered airway responsiveness.
Monitoring in vivo changes in lung microstructure with 3He MRI in Sendai virus-infected mice
Authors: Wang Wei , Nguyen Nguyet M. , Agapov Eugene , Holtzman Michael J. , Woods Jason C.
Published: March 2012

Recently, a Sendai virus (SeV) model of chronic obstructive lung disease has demonstrated an innate immune response in mouse airways that exhibits similarities to the chronic airway inflammation in human chronic obstructive pulmonary disease (COPD) and asthma, but the effect on distal lung parenchyma has not been investigated. The aim of our study is to image the time course and regional distribution of mouse lung microstructural changes in vivo after SeV infection. 1H and 3He diffusion magnetic resonance imaging (MRI) were successfully performed on five groups of C57BL/6J mice. 1H MR images provided precise anatomical localization and lung volume measurements. 3He lung morphometry was implemented to image and quantify mouse lung geometric microstructural parameters at different time points after SeV infection. 1H MR images detected the SeV-induced pulmonary inflammation in vivo; spatially resolved maps of acinar airway radius R, alveolar depth h, and mean linear intercept Lm were generated from 3He diffusion images. The morphometric parameters R and Lm in the infected group were indistinguishable from PBS-treated mice at day 21, increased slightly at day 49, and were increased with statistical significance at day 77 ( p = 0.02). Increases in R and Lm of infected mice imply that there is a modest increase in alveolar duct radius distal to airway inflammation, particularly in the lung periphery, indicating airspace enlargement after virus infection. Our results indicate that 3He lung morphometry has good sensitivity in quantifying small microstructural changes in the mouse lung and that the Sendai mouse model has the potential to be a valid murine model of COPD.
Aerobic exercise reduces neuronal responses in food reward brain regions
Authors: Evero Nero , Hackett Laura C. , Clark Robert D. , Phelan Suzanne , Hagobian Todd A.
Published: March 2012

Acute exercise suppresses ad libitum energy intake, but little is known about the effects of exercise on food reward brain regions. After an overnight fast, 30 (17 men, 13 women), healthy, habitually active (age = 22.2 ± 0.7 yr, body mass index = 23.6 ± 0.4 kg/m2, V̇o2peak = 44.2 ± 1.5 ml·kg−1·min−1) individuals completed 60 min of exercise on a cycle ergometer or 60 min of rest (no-exercise) in a counterbalanced, crossover fashion. After each condition, blood oxygen level-dependent responses to high-energy food, low-energy food, and control visual cues, were measured by functional magnetic resonance imaging. Exercise, compared with no-exercise, significantly ( P < 0.005) reduced the neuronal response to food (high and low food) cues vs. control cues in the insula (−0.37 ± 0.13 vs. +0.07 ± 0.18%), putamen (−0.39 ± 0.10 vs. −0.10 ± 0.09%), and rolandic operculum (−0.37 ± 0.17 vs. 0.17 ± 0.12%). Exercise alone significantly ( P < 0.005) reduced the neuronal response to high food vs. control and low food vs. control cues in the inferior orbitofrontal cortex (−0.94 ± 0.33%), insula (−0.37 ± 0.13%), and putamen (−0.41 ± 0.10%). No-exercise alone significantly ( P < 0.005) reduced the neuronal response to high vs. control and low vs. control cues in the middle (−0.47 ± 0.15%) and inferior occipital gyrus (−1.00 ± 0.23%). Exercise reduced neuronal responses in brain regions consistent with reduced pleasure of food, reduced incentive motivation to eat, and reduced anticipation and consumption of food. Reduced neuronal response in these food reward brain regions after exercise is in line with the paradigm that acute exercise suppresses subsequent energy intake.
Two-year citations of JAPPL original articles: evidence of a relative age effect
Authors: de Araújo Claudio Gil Soares , de Oliveira Bruno Ribeiro Ramalho , de Oliveira Brito Letícia Vargas , da Matta Thiago Torres , Viana Bruno Ferreira , de Souza Cintia Pereira , de Carvalho Guerreiro Renato , Slama Fabian Antonio , da Matta Mello Portugal Eduardo
Published: March 2012

Several indicators have been used to analyze scientific journals, with the impact factor and the number of citations in a 2-yr calendar time frame (2-YRC) being the most common factors. However, considering that the Journal of Applied Physiology (JAPPL) appears monthly and that calculations of these indicators are based on citations of papers published in previous years, we hypothesized that articles published at the beginning of the year would be cited more in the 2-YRC compared with those appearing in the last issues of the year, a phenomena known as a relative age effect. Our objective was to confirm the existence of a relative age effect in the 2-YRC for original articles published in JAPPL. From 2005 to 2008, a total of 1,726 original articles were published, according to the Web of Science, and 9,973 citations in 2-YRC, varying from 0 to 45, with a mean of 5.78 for individual papers. Although there were no differences in the number of original articles published in a given month ( P = 0.99), the 2-YRC varied considerably throughout the year, being higher for those earlier issues of the year, as shown by the linear regression analysis ( r2 = 0.76; P < 0.001). The 2-YRC began at 6.62 during the first 3 mo of the year, dropping by 10% at each 3-mo period. In summary, the longer an article has been out there, the more citations it collects. The relative age effect is a potential confounding variable for the assessment and interpretation of 2-YRC (using calendar years) from JAPPL original articles.
Prognostic and Predictive Significance of Plasma HGF and IL-8 in a Phase III Trial of Chemoradiation with or without Tirapazamine in Locoregionally Advanced Head and Neck Cancer
Authors: Le Quynh-Thu , Fisher Richard , Oliner Kelly S. , Young Richard J. , Cao Hongbin , Kong Christina , Graves Edward , Hicks Rodney J. , McArthur Grant A. , Peters Lester , O'Sullivan Brian , Giaccia Amato , Rischin Danny
Published: March 2012

Purpose: Hepatocyte growth factor (HGF) is a hypoxia-induced secreted protein that binds to cMet and regulates interleukin (IL)-8 expression. We evaluated the role of circulating HGF and IL-8 as prognostic and predictive factors for efficacy of tirapazamine (TPZ), a hypoxic cell cytotoxin.

Experimental Design: Patients with stages III to IV head and neck cancer were randomized to receive radiotherapy with cisplatin (CIS) or CIS plus TPZ (TPZ/CIS). Eligibility for the substudy included plasma sample availability for HGF and IL-8 assay by ELISA and no major radiation deviations (N = 498). Analyses included adjustment for major prognostic factors. p16INK4A staining (human papillomavirus surrogate) was carried out on available tumors. Thirty-nine patients had hypoxia imaging with 18F-fluoroazomycin arabinoside (18FAZA)–positron emission tomography.

Results: Elevated IL-8 level was associated with worse overall survival (OS) irrespective of treatment. There was an interaction between HGF and treatment arm (P = 0.053); elevated HGF was associated with worse OS in the control but not in the TPZ/CIS arm. Similar trends were observed in analyses restricted to p16INK4A-negative patients. Four subgroups defined by high and low HGF/IL-8 levels were examined for TPZ effect; the test for interaction with arm was P = 0.099. TPZ/CIS seemed to be beneficial for patients with high HGF and IL-8 but adverse for low HGF and high IL-8. Only HGF correlated with 18FAZA tumor standard uptake value.

Conclusions: IL-8 is an independent prognostic factor irrespective of treatment. There is an interaction between HGF and treatment arm. Certain subgroups based on IL-8/HGF levels seemed to do better with TPZ/CIS while others did worse, highlighting the complexity of hypoxia targeting in unselected patients. Clin Cancer Res; 18(6); 1798–807. ©2012 AACR.
Interleukin-27 Inhibits the Growth of Pediatric Acute Myeloid Leukemia in NOD/SCID/Il2rg−/− Mice
Authors: Zorzoli Alessia , Di Carlo Emma , Cocco Claudia , Ognio Emanuela , Ribatti Domenico , Ferretti Elisa , Dufour Carlo , Locatelli Franco , Montagna Daniela , Airoldi Irma
Published: March 2012

Purpose: Acute myeloid leukemia (AML) accounts for more than half of fatal cases in all pediatric leukemia patients; this observation highlights the need of more effective therapies. Thus, we investigated whether interleukin (IL)-27, an immunomodulatory cytokine, functions as an antitumor agent against pediatric AML cells.

Experimental Design: Expression of WSX-1 and gp130 on AML cells from 16 pediatric patients was studied by flow cytometry. Modulation of leukemia cell proliferation or apoptosis upon IL-27 treatment in vitro was tested by bromodeoxyuridine/propidium iodide (PI) and Ki67, or Annexin V/PI staining and flow cytometric analysis. The angiogenic potential of AML cells treated or not with IL-27 was studied by chorioallantoic membrane assay and PCR array. In vivo studies were carried out using nonobese diabetic/severe combined immunodeficient (NOD/SCID)/Il2rg−/− mice injected intravenously with five pediatric AML cell samples. Leukemic cells engrafted in PBS and IL-27–treated animals were studied by immunohistochemical/morphologic analysis and by PCR array for expression angiogenic/dissemination-related genes.

Results: We provided the first demonstration that (i) AML cells injected into NOD/SCID/Il2rg−/− mice gave rise to leukemia dissemination that was severely hampered by IL-27, (ii) compared with controls, leukemia cells harvested from IL-27–treated mice showed significant reduction of their angiogenic and spreading related genes, and (iii) similarly to what was observed in vivo, IL-27 reduced in vitro AML cell proliferation and modulated the expression of different genes involved in the angiogenic/spreading process.

Conclusion: These results provide an experimental rationale for the development of future clinical trials aimed at evaluating the toxicity and efficacy of IL-27. Clin Cancer Res; 18(6); 1630–40. ©2012 AACR.
A High-Throughput Panel for Identifying Clinically Relevant Mutation Profiles in Melanoma
Authors: Dutton-Regester Ken , Irwin Darryl , Hunt Priscilla , Aoude Lauren G. , Tembe Varsha , Pupo Gulietta M. , Lanagan Cathy , Carter Candace D. , O'Connor Linda , O'Rourke Michael , Scolyer Richard A. , Mann Graham J. , Schmidt Christopher W. , Herington Adrian , Hayward Nicholas K.
Published: March 2012

Success with molecular-based targeted drugs in the treatment of cancer has ignited extensive research efforts within the field of personalized therapeutics. However, successful application of such therapies is dependent on the presence or absence of mutations within the patient's tumor that can confer clinical efficacy or drug resistance. Building on these findings, we developed a high-throughput mutation panel for the identification of frequently occurring and clinically relevant mutations in melanoma. An extensive literature search and interrogation of the Catalogue of Somatic Mutations in Cancer database identified more than 1,000 melanoma mutations. Applying a filtering strategy to focus on mutations amenable to the development of targeted drugs, we initially screened 120 known mutations in 271 samples using the Sequenom MassARRAY system. A total of 252 mutations were detected in 17 genes, the highest frequency occurred in BRAF (n = 154, 57%), NRAS (n = 55, 20%), CDK4 (n = 8, 3%), PTK2B (n = 7, 2.5%), and ERBB4 (n = 5, 2%). Based on this initial discovery screen, a total of 46 assays interrogating 39 mutations in 20 genes were designed to develop a melanoma-specific panel. These assays were distributed in multiplexes over 8 wells using strict assay design parameters optimized for sensitive mutation detection. The final melanoma-specific mutation panel is a cost effective, sensitive, high-throughput approach for identifying mutations of clinical relevance to molecular-based therapeutics for the treatment of melanoma. When used in a clinical research setting, the panel may rapidly and accurately identify potentially effective treatment strategies using novel or existing molecularly targeted drugs. Mol Cancer Ther; 11(4); 888–97. ©2012 AACR.
Alexithymia and Severe Psychopathology: A Study on Psychiatric Inpatients
Authors: Picardi Angelo , Caroppo Emanuele , Porcelli Piero , Di Maria Giuseppe , Munittola Giampiero , Martinotti Giovanni
Published: March 2012

Background: The influence of severe psychopathology on alexithymic features has been poorly studied. This study aimed at investigating whether alexithymia is affected by the presence of psychosis and the severity of psychopathology. Sampling and Methods: 199 consecutive adult psychiatric inpatients, 60 of whom received a Structured Clinical Interview for DSM-IV Axis I diagnosis of psychotic disorder, were administered the 20-item Toronto Alexithymia Scale (TAS-20) and the 24-item Brief Psychiatric Rating Scale (BPRS). A random sample of 50 patients was reassessed at discharge. For all analyses, the Holm-Bonferroni correction was applied when appropriate to account for multiple testing. Results: The TAS-20 showed good internal consistency, acceptable relative stability, and good absolute stability both in patients with and without a psychotic disorder. Internal consistency and relative stability were higher in nonpsychotic than in psychotic patients. Mean TAS-20 score at admission did not differ between psychotic and nonpsychotic patients, and the TAS-20 score showed only a nonsignificant correlation (rho = 0.26) with BPRS score. Alexithymia at discharge was mostly explained (60%) by alexithymia at admission, while it was only modestly explained (4%) by BPRS scores at admission and discharge, with the latter being the only significant predictor. Changes in BPRS scores between admission and discharge explained a nonsignificant proportion (16%) of changes in TAS-20 scores and a moderate (21%) proportion of changes in difficulty identifying feelings. Conclusion: Despite some limitations, these findings suggest that the severity of psychopathology and the presence of a psychotic disorder only modestly influence alexithymic characteristics, and that the TAS-20 may provide a reliable assessment of alexithymia in patients with severe psychopathology, except the most severely impaired ones.
Specific Job Anxiety in Comparison to General Psychosomatic Symptoms at Admission, Discharge and Six Months after Psychosomatic Inpatient Treatment
Authors: Muschalla Beate , Linden Michael
Published: March 2012

Objective: Job anxiety is a severe problem in many patients with chronic mental disorders, as it usually results in specific participation problems in the workplace and long-term sick leave. The aim of this study was to explore the development of sick leave in dependence on general psychosomatic complaints and job anxiety from admission to a psychosomatic inpatient treatment until 6 months after discharge. Method: A convenience sample of 91 patients, suffering from multiple mental disorders, filled in self-rating questionnaires on job anxiety (Job Anxiety Scale) and on general psychosomatic symptom load (Symptom Checklist-90-Revised) at the beginning, the end, and 6 months after discharge from an inpatient psychosomatic treatment. Additionally, sick leave status and employment status were assessed before and 6 months after the treatment. Results: 15.4% of 91 patients were on sick leave before inpatient treatment and at follow-up (SS group), 20.9% were fit for work at intake and follow-up (FF group), 6.6% were fit for work initially and on sick leave later (FS group), and 57.1% on sick leave first and working at follow-up (SF group). In regard to general psychosomatic complaints, there were initially high scores on the SCL, a marked reduction during inpatient treatment, and a bouncing back to initial levels at follow-up for all 4 patient groups. SS and FS patients showed the highest scores at intake and follow-up. Concerning job anxiety, SS patients had the highest scores at all three assessments, while FF patients had significantly lower scores, with only low variation between assessments. SF patients started with comparatively high scores of job anxiety, which even increased before reentering work, but decreased in the follow-up period when they were confronted with work again. FS patients started low (like the FF patients) at intake, reduced their job anxiety further till discharge, but increased to higher scores at follow-up. Conclusions: General psychosomatic symptom load and job anxiety show a different course during treatment and are differently related to sick leave. General psychosomatic symptom load can be understood as a measure of the degree of the chronic illness status, whereas job anxiety reflects specific additional context-related problems, i.e. problems with work. A core finding is that job anxiety is related to work avoidance, but work exposure may reduce job anxiety. This has direct consequences for putting patients on sick leave or not.
Role of Substance Abuse Comorbidity and Personality on the Outcome of Depression in Bipolar Disorder: Harm Avoidance Influences Medium-Term Treatment Outcome
Authors: Mandelli Laura , Mazza Marianna , Di Nicola Marco , Zaninotto Leonardo , Harnic Desiree , Catalano Valeria , Tedeschi Daniela , Martinotti Giovanni , Bria Pietro , Janiri Luigi , Serretti Alessandro
Published: March 2012

Comorbid conditions are frequent in bipolar disorder (BD) and may complicate the treatment and course of illness. We investigated the role of substance use disorder (SUD), axis II personality disorders (PD) and continuous personality traits on the medium-term outcome (6 months) of treatment for bipolar depression. One hundred and thirty-nine BD patients meeting criteria for a depressive episode were included in the study. SUD and PD were diagnosed according to structured interviews. Personality dimensions were evaluated by the Temperament and Character Inventory. Depressive severity over time was evaluated by the Hamilton Rating Scale for Depression. Neither PD nor SUD influenced the outcome of depression. Variables independently associated with a poor outcome were a high baseline severity and high scores for the temperamental trait of Harm Avoidance. Though several limitations characterize the present study, neurotic personality traits seem to be associated with a slower recovery from depressive symptoms in BD, independently from their initial severity.
Baseline Serum Tryptase Levels and Adverse Reactions to Injection Specific Immunotherapy with Airborne Allergens: Is There a Relationship?
Authors: Asero R. , Farioli L. , Pastorello E.A.
Published: March 2012

Background: Elevated baseline serum tryptase levels are associated with severe systemic reactions following hymenoptera stings or venom immunotherapy. Little is known about baseline tryptase levels in patients with respiratory allergy and whether a relationship exists with systemic reactions induced by injection specific immunotherapy (SIT) with airborne allergens. The objective of this study was to measure tryptase levels in subjects with respiratory allergy and analyze the results in the light of tolerance/intolerance to injection SIT. Methods: Baseline serum tryptase levels were measured in 106 adults allergic to different airborne allergens and in 40 normal controls. Thirty-one patients underwent injection SIT, and 15 of these 31 experienced at least one SIT-induced systemic reaction. Results: Patients and normal controls showed similar median tryptase levels (2.98 vs. 3.13 ng/ml, respectively), although these were elevated in 6 patients (6%) versus 0 of 40 controls (0%). Tryptase levels did not differ between those patients with or without a history of systemic reactions (median 3.7 vs. 5.91 ng/ml, not significant). Three of 4 patients showing elevated tryptase levels belonged to the SIT-tolerant group. Elevated tryptase levels were not associated with specific allergens nor with distance from the specific pollen season. A bone marrow aspirate performed in the only patient with a history of systemic reactions following injection SIT and tryptase >11.4 ng/ml showed a normal morphology and phenotype. Conclusions: Unlike patients with hymenoptera venom allergy, in patients with respiratory allergy, elevated serum tryptase levels do not represent a risk factor for adverse reactions to SIT.
Estrogen Stimulates Th2 Cytokine Production and Regulates the Compartmentalisation of Eosinophils during Allergen Challenge in a Mouse Model of Asthma
Authors: Cai Yeping , Zhou Jiansheng , Webb Dianne C.
Published: March 2012

Background: The observation that asthma becomes more prevalent following puberty in females suggests estrogen potentiates the development of this disease. However, most studies examining the role of estrogen in rodent models of asthma are complicated by their reliance on ovariectomised mice in which hormones other than estrogen are also attenuated. Methods: We aimed to understand the influence of estrogen on allergic airway disease by using type I (tamoxifen) or type II (ICI 182,780) antagonists in female mice or delivering estradiol to male mice during aeroallergen challenge. Results: The antagonists showed that estrogen promoted both the mobilisation of bone marrow eosinophils and egression of eosinophils to the airway lumen. These findings were corroborated in male mice treated with estradiol, which increased eosinophil numbers in both blood and airways. Estrogen stimulated goblet cell hyperplasia and baseline lung resistance, but had little effect on the number of eosinophils in the bronchial submucosa or methacholine-induced airway hyperreactivity. Estrogen receptor α was expressed by CD4+ T cells from allergic mice, and estrogen promoted the production of IL-5 and IL-13, and suppressed the production of the eicosanoid 12-HETE by mediastinal lymph node cells. Conclusions: These data show that during aeroallergen challenge, estrogen stimulates Th2 cytokine production, which may be linked to its ability to suppress 12-HETE. Lung resistance at baseline, goblet cell hyperplasia and the compartmentalisation of eosinophils was also influenced by estrogen. However, estrogen does not play a major role in stimulating enhanced sensitivity to methacholine-induced lung resistance.
Neuroinflammation and Common Mechanism in Alzheimer’s Disease and Prion Amyloidosis: Amyloid-Associated Proteins, Neuroinflammation and Neurofibrillary Degeneration
Authors: Rozemuller Annemieke J.M. , Jansen Casper , Carrano Anna , van Haastert Elise S. , Hondius David , van der Vies Saskia M. , Hoozemans Jeroen J.M.
Published: March 2012

Background: In cases with a long (>1 year) clinical duration of prion disease, the prion protein can form amyloid deposits. These cases do not show accumulation of 4-kDa β-amyloid, which is observed in amyloid deposits in Alzheimer’s disease (AD). In AD, amyloid is associated with inflammation and neurofibrillary degeneration, and it is elusive whether prion amyloid is associated with these changes as well. Objectives: The presence of inflammation and neurofibrillary degeneration was evaluated in prion amyloidosis. Material and Methods: Cortical areas of variant Creutzfeldt-Jakob disease (CJD; n = 3), young sporadic CJD (n = 4), different Gerstmann-Sträussler-Scheinker’s disease patients (n = 5) and AD cases (n = 5) were examined using immunohistochemistry and specific stainings for amyloid. Results: In both AD and prion disease cases, which were negative for 4-kDa β-amyloid, parenchymal and vascular amyloid deposits were positive for amyloid-associated proteins such as complement protein and were associated with microglia clusters. Tau and ubiquitin were found near prion plaques in some of the Gerstmann-Sträussler-Scheinker’s disease and sporadic CJD cases and also near vascular prion amyloid deposits. In variant CJD cases, occasionally, microglia clustering was found in plaques but no ubiquitin or complement proteins and hardly tau protein. Conclusions: In both AD and prion disease amyloid formation, irrespective of the protein involved, there seems to be a neuroinflammatory response with secondary neurofibrillary degeneration.
Importance of Age for 3-Year Continuous Behavioral Obesity Treatment Success and Dropout Rate
Authors: Danielsson Pernilla , Svensson Viktoria , Kowalski Jan , Nyberg Gisela , Ekblom Örjan , Marcus Claude
Published: March 2012

Objective: To assess whether first year weight loss, age, and socioeconomic background correlate with the success rate of continuous long-term behavioral obesity treatment. Methods: In a 3-year longitudinal study, obese children (n = 684) were divided into three groups based on age at the start of treatment, age 6–9 years, 10–13 years, and 14–16 years. Results: The mean BMI standard deviation score (BMI-SDS) decline was age-dependent (p = 0.001), independently of adjustment for missing data: –1.8 BMI-SDS units in the youngest, –1.3 in the middle age group, and –0.5 in the oldest age group. SES and parental BMI status did not affect the results. 30% of the adolescents remained in treatment at year 3. There was only a weak correlation between BMI-SDS change after 1 and 3 years: r = 0.51 (p < 0.001). Among children with no BMI-SDS reduction during year 1 (n = 46), 40% had a clinically significantly reduced BMI-SDS after year 3. Conclusion: Behavioral treatment should be initiated at an early age to increase the chance for good results. Childhood obesity treatment should be continued for at least 3 years, regardless of the initial change in BMI-SDS.
Identification of the Translation Start Site of the Human Melanocortin 3 Receptor
Authors: Tarnow Patrick , Rediger Anne , Schulz Angela , Grüters Annette , Biebermann Heike
Published: March 2012

Background: The melanocortin-3-receptor (MC3R) is a G-protein coupled receptor participating in hypothalamic energy metabolism. So far, it was assumed that the translation of the human MC3R starts at the non-conserved first ATG, however, a second evolutionary conserved ATG is located 37 amino acids downstream. One frequent polymorphism, T6K, is located between these two ATGs. Methods: For characterization of the two potential start ATGs, COS-7 cells were transfected with plasmids encoding the longer and the shorter form of the human MC3R. For signal transduction properties, cAMP was measured. Cell surface expression was determined by using an ELISA method. The translational start point of the MC3R was investigated by a GFP-based method. Results: Signal transduction was comparable for the long and the short receptor form. Cell surface expression via aminoterminal hemagglutinin tag could only be detected in the shorter form, but not in the longer one. In our study we show that the translation of the human MC3R protein starts at the evolutionary conserved ATG codon which results in a shorter protein than previously assumed. Conclusion: The polymorphism T6K is not located in the coding region of the human MC3R and has no influence on translation initiation which makes an impact on body weight unlikely.
Physical Fitness Is Independently Related to Blood Leptin Concentration and Insulin Sensitivity Index in Male Subjects with Central Adiposity
Authors: Saafi Mohamed Ali , Frere-Meunier Delphine , Feasson Léonard , Boutahar Nadia , Denis Christian
Published: March 2012

Aim: To compare the maximal power output (MPO) of subjects presenting a central adiposity to those of controls and to study the links between plasma leptin or indices of insulin sensitivity (QUICKI) and physical fitness (PF). Methods: MPO was determined for 169 middle-aged men divided into two groups according to waist circumference (WC– < 94 cm, WC+ ≧ 94 cm) each subdivided in two subgroups with low and high PF (WC-L, WC-H, WC+L, WC+H) determined from the median MPO relative to fat free mass (3.06 W/kgFFM). Results: MPO (W/kgFFM) was lower in WC+ than in WC–. Expressed relative to fat mass, leptin was lower and QUICKI higher in WC– than in WC+. In WC+H, leptin and QUICKI were significantly less disturbed than in WC+L and were independently correlated to MPO (r = –0.36 and r = 0.32 respectively; p < 0.001). In WC+, when visceral perimeter was added to the analysis, the relationships MPO/leptin remained significant but not MPO/QUICKI. Conclusion: The low PF in subjects with abdominal obesity is independently linked to plasma leptin and insulin sensitivity even if leptin and insulin may share common pathways in their peripheral effects. Visceral adiposity participates to the link between MPO and QUICKI, but not between MPO and leptin.
Overweight Prevention Implemented by Primary School Teachers: A Randomised Controlled Trial
Authors: Brandstetter Susanne , Klenk Jochen , Berg Swantje , Galm Christoph , Fritz Michael , Peter Richard , Prokopchuk Dmytro , Steiner Ronald P. , Wartha Olivia , Steinacker Jürgen , Wabitsch Martin
Published: March 2012

Objective: To describe the effects of URMEL-ICE, a German school-based intervention for overweight prevention, on children’s BMI and other measures of fat mass. Methods: A cluster-randomised controlled design was used. The intervention which focused on physical activity, TV time and soft drink consumption was integrated into a second-grade curriculum and was implemented by classroom teachers themselves. It comprised 29 teaching lessons, 2 short exercise blocks per day and 6 family homework lessons. BMI was assessed as primary outcome measure, waist circumference and skinfold thickness as secondary outcomes. Data of 945 children were analysed. Results: Multivariate analyses adjusted for baseline values showed no statistically significant effect of the intervention on BMI, but on waist circumference (–0.85; 95% confidence interval (95% CI) –1.59 to –0.12) and subscapular skinfold thickness (–0.64; 95% CI –1.25 to –0.02). After additional adjustment for individual time lag between baseline and follow-up, these effects were reduced to –0.60 (95% CI –1.25 to 0.05) and –0.61 (95% CI –1.26 to 0.04) and lost their statistical significance. Conclusion: This study contributes to the field of randomised school-based studies on overweight prevention and shows that within a 1-year, integrated intervention no effect on BMI, but a tendency towards effects on fat mass can be achieved.