bluuebunny/crossref_metadata_2025 huggingface dataset:
The Alzheimer's Amyloid-Degrading Peptidase, Neprilysin: Can We Control It?
Authors: Nalivaeva N. N. , Belyaev N. D. , Zhuravin I. A. , Turner A. J.
Published: July 2012

The amyloid cascade hypothesis of Alzheimer's disease (AD) postulates that accumulation in the brain of amyloidβ-peptide (Aβ) is the primary trigger for neuronal loss specific to this pathology. In healthy brain, Aβlevels are regulated by a dynamic equilibrium between Aβrelease from the amyloid precursor protein (APP) and its removal by perivascular drainage or by amyloid-degrading enzymes (ADEs). During the last decade, the ADE family was fast growing, and currently it embraces more than 20 members. There are solid data supporting involvement of each of them in Aβclearance but a zinc metallopeptidase neprilysin (NEP) is considered as a major ADE. NEP plays an important role in brain function due to its role in terminating neuropeptide signalling and its decrease during ageing or after such pathologies as hypoxia or ischemia contribute significantly to the development of AD pathology. The recently discovered mechanism of epigenetic regulation of NEP by the APP intracellular domain (AICD) opens new avenues for its therapeutic manipulation and raises hope for developing preventive strategies in AD. However, consideration needs to be given to the diverse physiological roles of NEP. This paper critically evaluates general biochemical and physiological functions of NEP and their therapeutic relevance.
Enhanced Oxidative Stress and Physiological Damage in Daphnia magna by Copper in the Presence of Nano‐TiO2
Authors: Fan W. H. , Cui M. M. , Shi Z. W. , Tan C. , Yang X. P.
Published: July 2012

This study examines the potential hazard of an individual nanomaterial on the Cu biotoxicity to aquatic organisms. Daphnia magna in the absence or presence of nano‐TiO2 was exposed to Cu. Maintaining nano‐TiO2 at a safe concentration cannot eliminate its potential hazard. The biomarkers superoxide dismutase, catalase, and Na+/K+‐ATPase in D. magna were measured. Cu in the presence of nano‐TiO2 induced higher levels of oxidative stress and physiological damage because of the sorption of Cu. Nano‐TiO2 also caused Na+/K+‐ATPase inhibition possibly by impeding the Na+/K+ transfer channel. The correlations among the biomarkers, mortality, and accumulation further showed that the overloading reactive oxygen species generation caused by nano‐TiO2 contributed to deeper oxidative stress and physiological regulation, thereby causing greater toxic injury.
A Nonradioactive Fluorimetric SPE-Based Ceramide Kinase Assay Using NBD-C6-Ceramide
Authors: Van Overloop Helena , Van der Hoeven Gerd , Van Veldhoven Paul P.
Published: July 2012

Ceramide kinase (CERK) has been implicated in important cellular processes such as inflammation and apoptosis. Its activity is usually measured using radiolabeled ceramide or [γ-32P]-ATP, followed by extraction, thin-layer chromatography, and detection of the formed labeled ceramide-1-phosphate. To eliminate the use of radioactivity, we developed similarly but independently from the approach by Don and Rosen (2008), a fluorescence-based ceramide kinase assay, using N-[7-(4-nitrobenz-2-oxa-1,3-diazole)]-6-aminohexanoyl-sphingenine (NBD-C6-ceramide) as substrate. ItsKmvalue (4 μM) was comparable to that of N-hexanoyl-sphingenine (C6-ceramide). The produced fluorescent NBD-C6-ceramide-1-phosphate was captured by means of solid-phase extraction on an aminopropyl phase, resulting in a fast and sensitive CERK measurement. By performing this assay in a 96-well format, it is also suitable for high-throughput screening (HTS) to search for CERK modulators. A limited screen revealed that some protein kinase inhibitors (e.g., U-0126;IC504 μM) and ceramide analogues (e.g., fenretinide, AMG-9810;IC501.1 μM) affect CERKin vitro.
Contractile Activity Regulates Inducible Nitric Oxide Synthase Expression and NOiProduction in Cardiomyocytes via a FAK-Dependent Signaling Pathway
Authors: Chu Miensheng , Koshman Yevgeniya , Iyengar Rekha , Kim Taehoon , Russell Brenda , Samarel Allen M.
Published: July 2012

Intracellular nitric oxide (NOi) is a physiological regulator of excitation-contraction coupling, but is also involved in the development of cardiac dysfunction during hypertrophy and heart failure. To determine whether contractile activity regulates nitric oxide synthase (NOS) expression, spontaneously contracting, neonatal rat ventricular myocytes (NRVM) were treat with L-type calcium channel blockers (nifedipine and verapamil) or myosin II ATPase inhibitors (butanedione monoxime (BDM) and blebbistatin) to produce contractile arrest. Both types of inhibitors significantly reduced iNOS but not eNOS expression, and also reducedNOiproduction. Inhibiting contractile activity also reduced focal adhesion kinase (FAK) and AKT phosphorylation. Contraction-induced iNOS expression required FAK and phosphatidylinositol 3-kinase (PI(3)K), as both PF573228 and LY294002 (10 μM, 24 h) eliminated contraction-induced iNOS expression. Similarly, shRNAs specific for FAK (shFAK) caused FAK knockdown, reduced AKT phosphorylation at T308 and S473, and reduced iNOS expression. In contrast, shRNA-mediated knockdown of PYK2, the other member of the FAK-family of protein tyrosine kinases, had much less of an effect. Conversely, overexpression of a constitutively active form of FAK (CD2-FAK) or AKT (Myr-AKT) reversed the inhibitory effect of BDM on iNOS expression andNOiproduction. Thus, contraction-induced iNOS expression andNOiproduction in NRVM are mediated via a FAK-PI(3)K-AKT signaling pathway.
High‐Sensitive Sensor of Dopamine Based on Photoluminescence Quenching of Hierarchical CdS Spherical Aggregates
Authors: Sun Binjie , Wang Changzheng
Published: July 2012

Hierarchical CdS spherical aggregates have been fabricated by an assembling strategy starting from nanoparticles, which opens a general way to obtain hierarchical spherical aggregates of different types of materials. The hierarchical CdS spherical aggregates are of high porosity and high surface area, which give rise to unique photoluminescence properties. The desirable properties we report here will spur further developments of novel dopamine photoluminescence sensors based on the high surface area hierarchical CdS spherical aggregates fabricated with our unique assembling strategy. The novel dopamine photoluminescence sensor has a low detection limit of 1.0 × 10−8 M, which is much lower than those reported previously.
The Aggregation of Huntingtin and α-Synuclein
Authors: Chánez-Cárdenas María Elena , Vázquez-Contreras Edgar
Published: July 2012

Huntington’s and Parkinson’s diseases are neurodegenerative disorders associated with unusual protein interactions. Although the origin and evolution of these diseases are completely different, characteristic deposits of protein aggregates (huntingtin and α-synuclein resp.), are a common feature in both diseases. After these observations, many studies are performed with both proteins. Some of them try to understand the nature and driving forces of the aggregation process; others try to find a correlation between the genetic and failure in protein function. Finally with the combination of both approaches, it was proposed that possible strategies deal with pathologic aggregation. Unfortunately, if protein aggregation is a cause or a consequence of the neurodegeneration observed in these pathologies, it is still debatable. This paper describes the process of aggregation of two proteins: huntingtin and α synuclein. The characteristics of the aggregation reaction of these proteins have been followed with novel methods both in vivo and in vitro; these studies include both the combination with other proteins and the presence of various chemical compounds. The ultimate goal of this study was to summarize recent findings on protein aggregation and its possible role as a therapeutic target in neurodegenerative diseases and their role in biomaterial science.
Early Management Experience of Perforation after ERCP
Authors: Li Guohua , Chen Youxiang , Zhou Xiaojiang , Lv Nonghua
Published: July 2012

Background and Aim. Perforation after endoscopic retrograde cholangiopancreatography (ERCP) is a rare complication, but it is associated with significant mortality. This study evaluated the early management experience of these perforations.Patients and Methods. Between November 2003 and December 2011, a total of 8504 ERCPs were performed at our regional endoscopy center. Sixteen perforations (0.45%) were identified and retrospectively reviewed.Results. Nine of these 16 patients with perforations were periampullary, 3 duodenal, 1 gastric fundus, and 3 patients had a perforation of an afferent limb of a Billroth II anastomosis. All patients with perforations were recognized during ERCP by X-ray and managed immediately. One patient with duodenal perforation and three patients with afferent limb perforation received surgery, others received medical conservative treatment which included suturing lesion, endoscopic nasobiliary drainage (ENBD), endoscopic retrograde pancreatic duct drainage (ERPD), gastrointestinal decompression, fasting, broad-spectrum antibiotics, and so on. All patients with perforation recovered successfully.Conclusions. We found that: (1) the diagnosis of perforation during ERCP may be easy, but you must pay attention to it. (2) Most retroperitoneal perforations can recover with only medical conservative treatment in early phase. (3) Most peritoneal perforations need surgery unless you can close the lesion up under endoscopy in early phase.
Contemporary Patient Satisfaction Rates for Three-Piece Inflatable Penile Prostheses
Authors: Bernal Raymond M. , Henry Gerard D.
Published: July 2012

Among the many treatments for erectile dysfunction, implantation of a penile prosthesis has been associated with high patient satisfaction rates. Specifically, the placement of a three-piece inflatable penile prosthesis (IPP) confers the highest rates of satisfaction. We reviewed the literature over the past 20 years regarding satisfaction rates for penile prostheses, with a focus on patients who had undergone an initial IPP implantation for erectile dysfunction. In all, 194 articles were reviewed, and of these, nine met inclusion criteria for analysis and data collation. We determined contemporary satisfaction rates to reflect patients’ experiences with newer products and surgical approaches. Of importance, we noted that varied metrics were used to determine patient satisfaction, and overall satisfaction could not be precisely determined. Nevertheless, we found that patients in general were quite satisfied with their three-piece IPPs and restoration of sexual function. We also identified reasons for patient dissatisfaction and reviewed the literature to find ways by which satisfaction could be improved. Given the various means by which patient satisfaction was determined, future efforts should include standardized and validated questionnaires.
Primary Amoebic Meningoencephalitis in an Iranian Infant
Authors: Movahedi Zahra , Shokrollahi Mohammad Reza , Aghaali Mohammad , Heydari Hosein
Published: July 2012

Introduction.Naegleria fowleri, a free living amoeba, can cause devastating and deadly diseases in humans. This is the first report of primary amoebic meningoencephalitis from Iran.Case report. A five-month-old male infant presented with the history of fever and eye gaze for three days, after beginning of bacterial meningitis, a plain and contrast CT revealed communicated hydrocephalus. In the repeat of CSF analysis on microscopic examination of wet preparation of CSF,Naegleria Fowleriwas seen. Then, Amphotericin B and Rifampin were started. On followup, two months later, the patient was totally asymptomatic.Conclusion. Though occurrence of PAM is rare, this unusual disease has grave prognosis, so infection with free living amoebas must be considered in differential diagnosis of pediatric patients of purulent meningitis without evidence of bacteria on Gram’s stain and imaging findings, nonspecific brain edema on CT or hydrocephalus even without history of contact.
Recent Findings on the Role of Gelatinases (Matrix Metalloproteinase-2 and -9) in Osteoarthritis
Authors: Galasso Olimpio , Familiari Filippo , De Gori Marco , Gasparini Giorgio
Published: July 2012

Several studies dealing with the pathomechanisms of OA refer to MMP-1, -3, -7, -8, and -13 whereas a smaller number of investigations have pointed out the pathogenic role of gelatinases in OA. These gelatinases are best known for their involvement in pulmonary, myocardial, and neoplastic disease but they are emerging as important proteases implicated in the OA progression. This paper highlights the role of the gelatinases as emerging factors in OA pathogenesis through the regulation of subchondral bone resorption and microvascular invasion. The most significant new findings over the last year that add to our knowledge of the activity of these proteins in OA have been reported.
Combining Diffusion and Grey Models Based on Evolutionary Optimization Algorithms to Forecast Motherboard Shipments
Authors: Wang Fu-Kwun , Hsiao Yu-Yao , Chang Ku-Kuang
Published: July 2012

It is important for executives to predict the future trends. Otherwise, their companies cannot make profitable decisions and investments. The Bass diffusion model can describe the empirical adoption curve for new products and technological innovations. The Grey model provides short‐term forecasts using four data points. This study develops a combined model based on the rolling Grey model (RGM) and the Bass diffusion model to forecast motherboard shipments. In addition, we investigate evolutionary optimization algorithms to determine the optimal parameters. Our results indicate that the combined model using a hybrid algorithm outperforms other methods for the fitting and forecasting processes in terms of mean absolute percentage error.
Optimization of Fermentation Medium for the Production of Glucose Isomerase UsingStreptomyces sp. SB-P1
Authors: Bhasin Sheetal , Modi H. A.
Published: July 2012

The combination of medium ingredients has a profound influence on the metabolic pathways running in the microorganism which regulates the production of numerous metabolites. Glucose isomerase (GI), an enzyme with huge potential in the market, can isomerise glucose into fructose. GI is used widely for the production of High-Fructose Corn Syrup (HFCS). HFCS is used as a sweetener in food and pharmaceutical industries.Streptomycesare well-known producers of numerous enzymes including glucose isomerase. An array of 75 isolates was screened for the production of glucose isomerase. The isolateStreptomyces sp. SB-P1was found to produce maximum amount of extracellular GI. Sucrose and raffinose among pure carbon sources and corn cob and wheat husk among crude agro residues were found to yield high enzyme titers. Potassium nitrate among pure nitrogen sources and soy residues among crude sources gave maximum production. Quantitative effect of carbon, nitrogen, and inducer on GI was also determined. Plackett-Burman design was used to study the effect of different medium ingredients. Sucrose and xylose as carbon sources and peptone and soy residues as nitrogen sources proved to be beneficial for GI production.
Assessment of a Standardized ROS Production Profile in Humans by Electron Paramagnetic Resonance
Authors: Mrakic-Sposta Simona , Gussoni Maristella , Montorsi Michela , Porcelli Simone , Vezzoli Alessandra
Published: July 2012

Despite the growing interest in the role of reactive oxygen species (ROS) in health and disease, reliable quantitative noninvasive methods for the assessment of oxidative stress in humans are still lacking. EPR technique, coupled to a specific spin probe (CMH: 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine) is here presented as the method of choice to gain a direct measurement of ROS in biological fluids and tissues. The study aimed at demonstrating that, differently from currently available “a posteriori” assays of ROS-induced damage by means of biomolecules (e.g., proteins and lipids) spin-trapping EPR provides direct evidence of the “instantaneous” presence of radical species in the sample and, as signal areas are proportional to the number of excited electron spins, lead to absolute concentration levels. Using a recently developed bench top continuous wave system (e-scan EPR scanner, Bruker) dealing with very low ROS concentration levels in small (50 μL) samples, we successfully monitored rapid ROS production changes in peripheral blood of athletes after controlled exercise and sedentary subjects after antioxidant supplementation. The correlation between EPR results and data obtained by various enzymatic assays (e.g., protein carbonyls and thiobarbituric acid reactive substances) was determined too. Synthetically, our method allows reliable, quick, noninvasive quantitative determination of ROS in human peripheral blood.
MicroRNAs Regulate Tumor Angiogenesis Modulated by Endothelial Progenitor Cells
Authors: Plummer Prue N. , Freeman Ruth , Taft Ryan J. , Vider Jelena , Sax Michael , Umer Brittany A. , Gao Dingcheng , Johns Christopher , Mattick John S. , Wilton Stephen D. , Ferro Vito , McMillan Nigel A.J. , Swarbrick Alexander , Mittal Vivek , Mellick Albert S.
Published: July 2012

Bone marrow–derived endothelial progenitor cells (EPC) contribute to the angiogenesis-dependent growth of tumors in mice and humans. EPCs regulate the angiogenic switch via paracrine secretion of proangiogenic growth factors and by direct luminal incorporation into sprouting nascent vessels. miRNAs have emerged as key regulators of several cellular processes including angiogenesis; however, whether miRNAs contribute to bone marrow–mediated angiogenesis has remained unknown. Here, we show that genetic ablation of miRNA-processing enzyme Dicer, specifically in the bone marrow, decreased the number of circulating EPCs, resulting in angiogenesis suppression and impaired tumor growth. Furthermore, genome-wide deep sequencing of small RNAs revealed tumor EPC-intrinsic miRNAs including miR-10b and miR-196b, which have been previously identified as key regulators of HOX signaling and adult stem cell differentiation. Notably, we found that both miR-10b and miR-196b are responsive to vascular endothelial growth factor stimulation and show elevated expression in human high-grade breast tumor vasculature. Strikingly, targeting miR-10b and miR-196b led to significant defects in angiogenesis-mediated tumor growth in mice. Targeting these miRNAs may constitute a novel strategy for inhibiting tumor angiogenesis. Cancer Res; 73(1); 341–52. ©2012 AACR.
Ikk4a/Arf Inactivation with Activation of the NF-κB/IL-6 Pathway Is Sufficient to Drive the Development and Growth of Angiosarcoma
Authors: Yang Jinming , Kantrow Sara , Sai Jiqing , Hawkins Oriana E. , Boothby Mark , Ayers Gregory D. , Young Eric D. , Demicco Elizabeth G. , Lazar Alexander J. , Lev Dina , Richmond Ann
Published: July 2012

Although human angiosarcoma has been associated frequently with mutational inactivation of the tumor suppressor gene Ink4a/Arf, the underlying mechanisms have not been delineated. Here we report that malignant angiosarcoma is associated with high levels of RelA/NF-κB and IL-6 in contrast to normal vessels or benign hemagiomas. Studies of Ink4a/Arf deficient mice not only recapitulate genetic traits observed in human angiosarcoma, but also unveil a possible therapeutic link comprised of the NF-kB/IL-6/Stat3 signaling axis. In Ink4a/Arf−/− cells, NF-κB controlled Stat3 signaling by transcriptionally controlling the expression of IL-6, gp130, and Jak2. Further, IL-6 mediated Stat3 signaling through the sIL-6R. Inhibition of Ikkβ solely in myeloid cells was insufficient to block angiosarcoma development; in contrast, systemic inhibition of Ikkβ, IL-6, or Stat3 markedly inhibited angiosarcoma growth. Our findings offer clinical implications for targeting the NF-kB/IL-6/STAT3 pathway as a rational strategy to treat angiosarcoma. Cancer Res; 72(18); 4682–95. ©2012 AACR.
B-Raf Activation Cooperates with PTEN Loss to Drive c-Myc Expression in Advanced Prostate Cancer
Authors: Wang Jingqiang , Kobayashi Takashi , Floc'h Nicolas , Kinkade Carolyn Waugh , Aytes Alvaro , Dankort David , Lefebvre Celine , Mitrofanova Antonina , Cardiff Robert D. , McMahon Martin , Califano Andrea , Shen Michael M. , Abate-Shen Cory
Published: July 2012

Both the PI3K → Akt → mTOR and mitogen-activated protein kinase (MAPK) signaling pathways are often deregulated in prostate tumors with poor prognosis. Here we describe a new genetically engineered mouse model of prostate cancer in which PI3K-Akt-mTOR signaling is activated by inducible disruption of PTEN, and extracellular signal-regulated kinase 1/2 (ERK1/2) MAPK signaling is activated by inducible expression of a BRAFV600E oncogene. These tissue-specific compound mutant mice develop lethal prostate tumors that are inherently resistant to castration. These tumors bypass cellular senescence and disseminate to lymph nodes, bone marrow, and lungs where they form overt metastases in approximately 30% of the cases. Activation of PI3K → Akt → mTOR and MAPK signaling pathways in these prostate tumors cooperate to upregulate c-Myc. Accordingly, therapeutic treatments with rapamycin and PD0325901 to target these pathways, respectively, attenuate c-Myc levels and reduce tumor and metastatic burden. Together, our findings suggest a generalized therapeutic approach to target c-Myc activation in prostate cancer by combinatorial targeting of the PI3K → Akt → mTOR and ERK1/2 MAPK signaling pathways. Cancer Res; 72(18); 4765–76. ©2012 AACR.
A Phase I Trial of Dasatinib, an Src-Family Kinase Inhibitor, in Combination with Paclitaxel and Carboplatin in Patients with Advanced or Recurrent Ovarian Cancer
Authors: Secord Angeles Alvarez , Teoh Deanna K. , Barry William T. , Yu Miao , Broadwater Gloria , Havrilesky Laura J. , Lee Paula S. , Berchuck Andrew , Lancaster Johnathan , Wenham Robert M.
Published: July 2012

Purpose: We conducted a phase I study of dasatinib, an oral SRC-family tyrosine kinase inhibitor, in combination with paclitaxel and carboplatin in the treatment of advanced and recurrent epithelial ovarian cancer.

Experimental Design: The primary objective was to determine the maximum tolerated dose (MTD). Secondary objectives included defining toxicity, response rate (RR), pharmacokinetics, and pharmacodynamics. Using a “3+3” design, cohorts of three to six patients received paclitaxel (175 mg/m2) and carboplatin (AUC 6) every 3 weeks with escalating doses of dasatinib (100, 120, and 150 mg daily), followed by an eight-patient expansion cohort.

Results: Twenty patients were enrolled between June 2007 and December 2009. The median age was 61 years (range: 42–82) with a median of 2 prior regimens (range: 0–6), and 71% had platinum-sensitive disease. There were three to six patients in each cohort, and eight in the expansion cohort. Pharmacokinetics were observed over the first two cycles of therapy. One DLT was observed in the 100 mg dasatinib cohort (grade 3 myalgia). Other toxicities in all cycles included neutropenia (95% grade 3–4; 91% in the 150 mg dosing cohort), thrombocytopenia (35% grade 3–4), and fatigue (10% grade 3). The RR was 40% [three complete responses, (15%); five partial responses, (25%)],10 patients (50%) had stable disease, and two were not evaluable. The PFS6-month actuarial estimate was 86%. The median PFS and OS were 7.8 and 16.2 months, respectively.

Conclusions: Due to the high incidence of myelosuppression with subsequent cycles, the recommended phase II dose of dasatinib is 150 mg daily in combination with paclitaxel and carboplatin. The combination was safe with evidence of clinical activity. Clin Cancer Res; 18(19); 5489–98. ©2012 AACR.
Myeloid Cells Obtained from the Blood but Not from the Tumor Can Suppress T-cell Proliferation in Patients with Melanoma
Authors: Gros Alena , Turcotte Simon , Wunderlich John R. , Ahmadzadeh Mojgan , Dudley Mark E. , Rosenberg Steven A.
Published: July 2012

Purpose: Myeloid-derived suppressor cells (MDSC) have emerged as an immune-regulatory cell type that is expanded in tumor-bearing mice, but less is known about their immune-suppressive role in patients with cancer.

Experimental Design: To study the importance of MDSC in patients with melanoma, we characterized the frequency, phenotype, and suppressive function of blood myeloid-derived cells and tumor-infiltrating myeloid cells in 26 freshly resected melanomas.

Results: Blood and tumor-infiltrating myeloid cells (Lin− CD11b+) could be phenotypically and morphologically classified into monocytes/macrophages, neutrophils, eosinophils, and immature myeloid cells according to marker expression (CD14+, CD14− CD15hi, CD14− CD15int, and CD14− CD15−, respectively). In contrast to the expansion of MDSC reported in tumor-bearing mice, we found no differences in the frequency and phenotype of myeloid subsets in the blood of patients with melanoma compared with healthy donors. Myeloid cells represented 12% of the live cells in the melanoma cell suspensions, and were phenotypically diverse with high tumor-to-tumor variability. Interestingly, a positive association was found between the percentage of Tregs and granulocytic cells (Lin− CD11b+ CD14−CD15+) infiltrating melanoma tumors. However, melanoma-infiltrating myeloid cells displayed impaired suppression of nonspecific T-cell proliferation compared with peripheral blood myeloid cells, in which monocytes and eosinophils were suppressive.

Conclusions: Our findings provide a first characterization of the nature and suppressive function of the melanoma myeloid infiltrate and indicate that the suppressive function of MDSC in patients with melanoma seems far less than that based on murine tumor models. Clin Cancer Res; 18(19); 5212–23. ©2012 AACR.
DNA Damage Repair Pathways in Cancer Stem Cells
Authors: Maugeri-Saccà Marcello , Bartucci Monica , De Maria Ruggero
Published: July 2012

The discovery of tumor-initiating cells endowed with stem-like features has added a further level of complexity to the pathobiology of neoplastic diseases. In the attempt of dissecting the functional properties of this uncommon cellular subpopulation, investigators are taking full advantage of a body of knowledge about adult stem cells, as the “cancer stem cell model” implies that tissue-resident stem cells are the target of the oncogenic process. It is emerging that a plethora of molecular mechanisms protect cancer stem cells (CSC) against chemotherapy- and radiotherapy-induced death stimuli. The ability of CSCs to survive stressful conditions is correlated, among others, with a multifaceted protection of genome integrity by a prompt activation of the DNA damage sensor and repair machinery. Nevertheless, many molecular-targeted agents directed against DNA repair effectors are in late preclinical or clinical development while the identification of predictive biomarkers of response coupled with the validation of robust assays for assessing biomarkers is paving the way for biology-driven clinical trials. Mol Cancer Ther; 11(8); 1627–36. ©2012 AACR.
Combined Therapy with Mutant-Selective EGFR Inhibitor and Met Kinase Inhibitor for Overcoming Erlotinib Resistance in EGFR-Mutant Lung Cancer
Authors: Nakagawa Takayuki , Takeuchi Shinji , Yamada Tadaaki , Nanjo Shigeki , Ishikawa Daisuke , Sano Takako , Kita Kenji , Nakamura Takahiro , Matsumoto Kunio , Suda Kenichi , Mitsudomi Tetsuya , Sekido Yoshitaka , Uenaka Toshimitsu , Yano Seiji
Published: July 2012

Although the EGF receptor tyrosine kinase inhibitors (EGFR-TKI) erlotinib and gefitinib have shown dramatic effects against EGFR mutant lung cancer, patients become resistant by various mechanisms, including gatekeeper EGFR-T790M mutation, Met amplification, and HGF overexpression, thereafter relapsing. Thus, it is urgent to develop novel agents to overcome EGFR-TKI resistance. We have tested the effects of the mutant-selective EGFR-TKI WZ4002 and the mutant-selective Met-TKI E7050 on 3 EGFR mutant lung cancer cell lines resistant to erlotinib by different mechanisms: PC-9/HGF cells with an exon 19 deletion, H1975 with an L858R mutation, and HCC827ER with an exon 19 deletion, with acquired resistance to erlotinib because of HGF gene transfection, gatekeeper T790M mutation, and Met amplification, respectively. WZ4002 inhibited the growth of H1975 cells with a gatekeeper T790M mutation, but did not inhibit the growth of HCC827ER and PC-9/HGF cells. HGF triggered the resistance of H1975 cells to WZ4002, whereas E7050 sensitized HCC827ER, PC-9/HGF, and HGF-treated H1975 cells to WZ4002, inhibiting EGFR and Met phosphorylation and their downstream molecules. Combined treatment potently inhibited the growth of tumors induced in severe-combined immunodeficient mice by H1975, HCC827ER, and PC-9/HGF cells, without any marked adverse events. These therapeutic effects were associated with the inhibition of EGFR and Met phosphorylation in vivo. The combination of a mutant-selective EGFR-TKI and a Met-TKI was effective in suppressing the growth of erlotinib-resistant tumors caused by gatekeeper T790M mutation, Met amplification, and HGF overexpression. Further evaluations in clinical trials are warranted. Mol Cancer Ther; 11(10); 2149–57. ©2012 AACR.
Nox4 Mediates Hypoxia-Stimulated Myofibroblast Differentiation in Nasal Polyp-Derived Fibroblasts
Authors: Moon You-Mi , Kang Hee Joon , Cho Jung-Sun , Park Il-Ho , Lee Heung-Man
Published: July 2012

Background: Chronic hypoxia is associated with remodeling in various organs. Reactive oxygen species (ROS) derived from NADPH oxidases (Nox), and transforming growth factor-β1 (TGF-β1) have been implicated in the pathogenesis of hypoxia-induced remodeling. The aims of this study were to determine in hypoxia-stimulated nasal polyp-derived fibroblasts (NPDF) the effect of hypoxia on the differentiation of myofibroblasts, the role of ROS, the major Nox homolog mediating myofibroblast differentiation, and the role of TGF-β1. Methods: Eight primary cultures of NPDF were established from nasal polyps, which were incubated under hypoxic conditions. Reverse transcription polymerase chain reaction for αSMA, Nox1, Nox3, Nox4, Nox5, and fibronectin mRNA was performed. Western blotting for α-SMA and fibronectin was done. ROS production was detected using a fluorometer. NPDF were pretreated with ROS scavengers and transfected with siNox4. The TGF-β1 protein level was measured by ELISA. The effect of treatment with TGF-β1 type I tyrosine kinase inhibitor SB431542 on myofibroblast differentiation was observed. Results: Hypoxic stimulation of NPDF significantly increased α-SMA and fibronectin mRNA and protein expression. ROS production was increased by hypoxia, and ROS scavengers inhibited myofibroblast differentiation. Nox4 mRNA was the only Nox homolog increased by hypoxia. Transfection with siNox4 inhibited myofibroblast differentiation. TGF-β1 was secreted endogenously by hypoxic NPDF. SB431542 significantly inhibited myofibroblast differentiation. Conclusions: Hypoxia induces myofibroblast differentiation of NPDF through a signaling pathway involving Nox4-dependent ROS generation and TGF-β1. Therapies targeting Nox4 may be effective against remodeling of nasal polyps.
Dilated Fourth Ventricle in Fetuses with Trisomy 18, Trisomy 13 and Triploidy at 11–13 Weeks’ Gestation
Authors: Loureiro Teresa , Ferreira Ana Fatima A. , Ushakov Fred , Montenegro Nuno , Nicolaides Kypros H.
Published: July 2012

Objective: To determine if in fetuses with aneuploidies the diameter of the fourth cerebral ventricle at 11–13 weeks’ gestation is different from euploid fetuses. Methods: The fourth ventricle at 11–13 weeks’ gestation was assessed in 62 cases of trisomy 21, 32 of trisomy 18, 10 of trisomy 13, and 12 of triploidy and compared to 410 normal euploid fetuses. Transvaginal sonography was carried out and 3D brain volumes were acquired. The fetal head was assessed in an axial plane and the diameter of the fourth ventricle was measured. Values in aneuploid and euploid fetuses were compared. Results: The diameter of the fourth ventricle in trisomy 18, trisomy 13 and triploidy, but not in trisomy 21, was significantly higher than in euploid fetuses. In the euploid fetuses the median diameter of the fourth ventricle was 1.9 mm and the 95th percentile was 2.5 mm. The measurements were above the median and the 95th percentile in 25 (78.1%) and 17 (53.1%) cases of trisomy 18, in 10 (100%) and 8 (80.0%) of trisomy 13, and in 10 (83.3%) and 10 (83.3%) of triploidy. Conclusions: In trisomy 18, trisomy 13 and triploidy the diameter of the fourth ventricle at 11–13 weeks’ gestation is increased.
Allergic Airway Inflammation in Mice Deficient for the Antigen-Processing Protease Cathepsin E
Authors: Pilzner Carolin , Bühling Frank , Reinheckel Thomas , Chwieralski Caroline , Rathinasamy Anchana , Lauenstein Hans-Dieter , Wex Thomas , Welte Tobias , Braun Armin , Groneberg David A.
Published: July 2012

Background: Allergic asthma is a Th2-type chronic inflammatory disease of the lung. It is characterized by infiltration of eosinophils, neutrophils, mast cells and T lymphocytes into the airways. Th2 cytokines like interleukin (IL)-4, IL-5 and chemokines like eotaxin are increased in the asthmatic response. The processing and presentation of exogenous antigens is important in the sensitization to an allergen. Cathepsin E (Ctse) is an intracellular aspartic endoprotease which is expressed in immune cells like dendritic cells (DCs). It was found to play an essential role in the processing and presentation of ovalbumin (OVA). The aim of the present study was to investigate the inhibition of Ctse in two different experimental models of allergic airway inflammation. Methods: Ctse wild-type (Ctse+/+) and Ctse-deficient (Ctse–/–) bone marrow-derived DCs (BMDCs) were pulsed with OVA/OVA peptide and cocultured with OVA transgenic T II (OT II) cells whose proliferation was subsequently analyzed. Two different in vivo asthma models with Ctse+/+ and Ctse–/– mice were performed: an acute OVA-induced and a subchronic Phleum pratense-induced airway inflammation. Results: Proliferation of OT II cells was decreased when cocultured with BMDCs of Ctse–/– mice as compared to cells cocultured with BMDCs of Ctse+/+ mice. In vivo, Ctse deficiency led to reduced lymphocyte influx after allergen sensitization and challenge in both investigated airway inflammation models, compared to their control groups. Conclusion: Ctse deficiency leads to a reduced antigen presentation in vitro. This is followed by a distinct effect on lymphocyte influx in states of allergic airway inflammation in vivo.
Montelukast Inhibits Leukotriene Stimulation of Human Dendritic Cells in vitro
Authors: Ilarraza Ramses , Wu Yingqi , Adamko Darryl J.
Published: July 2012

Background: Leukotrienes are potent inflammatory mediators which modulate immune responses and induce bronchoconstriction in susceptible individuals. Montelukast (MK) is a leukotriene receptor (CysLT1) antagonist that has been shown to prevent exacerbation of asthma. Considering the plethora of potential cellular targets for MK, specific mechanisms for its therapeutic action are still not fully understood. In vitro, we determined whether human dendritic cell function could be affected by leukotriene C4 (LTC4) treatment and whether MK had potential in modulating this response. We also studied the effect of LTC4 in the context of response to an airway virus (respiratory syncytial virus, RSV). Methods: Human monocyte-derived dendritic cells (moDCs) exposed to LTC4, MK, or both, were cocultured with autologous T cells, with or without RSV. The effects of LTC4 and MK on cell function were determined by ELISA and proliferation assays. Results: Both moDCs and their precursors – monocytes – express LTC4 receptor CysLT1, making them potential targets for MK. moDCs cultured with LTC4 release the eosinophil chemoattractant RANTES (CCL5) and induce greater T cell proliferation. Both were blocked by the presence of MK. MK treatment, albeit anti-inflammatory, did not interfere with the moDC-dependent T cell-proliferative responses induced by RSV. Conclusions: LTC4, chronically present in the airways of asthma patients, could induce an exaggerated inflammatory response to airway infection via dendritic cell activation, which would be prevented by MK. Our study provides additional insight into the mechanisms of action of this leukotriene receptor antagonist.
Determinants of Pulmonary Hypertension in Patients with Beta-Thalassemia Major and Normal Ventricular Function
Authors: Vlahos Antonios P. , Koutsouka Frideriki P. , Papamichael Nikolaos D. , Makis Alexandros , Baltogiannis Giannis G. , Athanasiou Eleni , Chaliasos Nikolaos , Bourantas Konstantinos L. , Kolettis Theofilos M.
Published: July 2012

Background/Aims: We sought to define the incidence and predictive factors of pulmonary hypertension in β-thalassemia major. Methods: We studied 27 consecutive patients (19 male, 38 ± 9 years of age) with β-thalassemia major. All the patients had normal (left and right) ventricular (systolic and diastolic) function and underwent echocardiographic assessment of pulmonary artery systolic pressure. Univariate regression and discriminant function analyses were used to identify predictive factors of pulmonary hypertension. Results: Pulmonary hypertension was observed in 18.5% of the patients, but clinically significant disease was detected in only 3.7%. A total of 14 (51.8%) patients had been receiving a combined administration of deferoxamine and deferiprone for 7.0 ± 1.3 years. Amidst a large number of variables examined, ferritin levels and delayed onset of chelation therapy were the only predictors of pulmonary hypertension. Conclusion: Pulmonary hypertension in β-thalassemia major is relatively infrequent and generally mild due to improved chelation therapy. The role of hemochromatosis in pulmonary hypertension development merits further study.
The Use of Complementary and Alternative Medicine by Pediatric Food-Allergic Patients in Japan
Authors: Nakano Taiji , Shimojo Naoki , Okamoto Yoshitaka , Ebisawa Motohiro , Kurihara Kazuyuki , Hoshioka Akira , Yamaguchi Koichi , Ito Komei , Fujisawa Takao , Kameda Makoto , Suehiro Yutaka , Ogura Hideo , Shibata Rumiko , Suzuki Shuichi , Takahashi Yutaka , Ikeda Masanori , Kohno Yoichi
Published: July 2012

Background: In developed countries, increasing food allergy prevalence and concern regarding food allergies have been reported. Although the use of complementary and alternative medicine (CAM) for the treatment of allergic diseases has increased in some Western countries, the actual proportion and patterns of CAM use for pediatric food allergies in Japan are still unknown. Methods: Fourteen allergy centers in Japan participated in the study using a questionnaire survey regarding the use of CAM by pediatric patients. A diagnosis of food allergy was made at each hospital by pediatric allergists. Results: Surveys were completed by parents/guardians, and data were collected for a total of 962 pediatric food-allergic patients. Overall, 8.4% of the participants used CAM to treat a food allergy. The major CAM therapies used were herbal teas (22.2%), including several Japanese herbal teas, Chinese herbal medicine (18.5%) and lactic acid bacteria (16%). Among the participants using CAM to treat food allergy, 13.6% thought that the CAM being used was very effective, while 11.1% of participants thought that CAM caused some type of side effect. Conclusions: Our study is the first large-scale national survey regarding the use of CAM in pediatric patients with food allergies in Japan. Unlike in the USA, which has a higher rate of CAM use (17%), approximately 8.4% of food-allergic patients used CAM in Japan. Interestingly, the major types of CAM used in Japan differed from those used in the USA. Cultural differences and food customs may affect the use of CAM.
Specific Immunotherapy Normalizes Tryptophan Concentrations in Patients with Allergic Rhinitis
Authors: Kofler H. , Kurz K. , Grander G. , Fuchs D.
Published: July 2012

Background/Aims: An immune shift towards Th2-type immunity seems to be critical in the pathogenesis of allergic asthma and rhinitis. In a previous study, we found higher serum tryptophan concentrations in patients with seasonal tree or grass pollen rhinoconjunctivitis who underwent specific immunotherapy (SCIT) than in controls, and those with the highest levels at baseline responded less well to SCIT. In the present study, we examined whether ‘booster immunotherapy’ after cessation of SCIT had any influence on tryptophan metabolism during follow-up. Methods: Serum concentrations of tryptophan, kynurenine and neopterin were assayed in 19 patients (mean age: 26.2 years; 6 females) allergic to grass and/or tree pollen before and after they had received a booster immunotherapy with 4 injections of an allergoid vaccine (Pollinex Quattro; Bencard Vienna, Austria) over 8 ± 3 months outside the pollen season. Results: Serum tryptophan and kynurenine concentrations decreased after booster immunotherapy (mean ± SD, before immunotherapy: 81.1 ± 14.2 µmol/l, after immunotherapy: 61.4 ± 20.9 µmol/l and before immunotherapy: 2.25 ± 0.44, after immunotherapy: 1.69 ± 0.70 µmol/l, respectively; both p < 0.01); this was especially true in those responders who also tended to have lower baseline kynurenine concentrations as compared with nonresponders (p = 0.05). Finally, a correlation between changes in tryptophan metabolism and neopterin concentrations was observed after immunotherapy. Conclusions: The decrease in tryptophan and kynurenine concentrations following booster immunotherapy in hay fever patients strengthens the hypothesis that tryptophan metabolism might be involved in the course of allergic responses. However, it is still unclear whether the abnormal tryptophan metabolism in pollinosis patients is related to indoleamine 2,3-dioxygenase and/or to a specific cytokine background.
A Population-Based Study of Fish Allergy in the Philippines, Singapore and Thailand
Authors: Connett Gary James , Gerez Irvin , Cabrera-Morales Elizabeth Ann , Yuenyongviwat Araya , Ngamphaiboon Jarungchit , Chatchatee Pantipa , Sangsupawanich Pasuree , Soh Shu-E , Yap Gaik-Chin , Shek Lynette Pei-Chi , Lee Bee-Wah
Published: July 2012

Background: Fish allergy is the third most common food allergy after milk and egg in parts of Europe, but there is little data about prevalence in South East Asia where it is an important part of regular diets. Objective: We aimed to obtain an estimate of the population prevalence of fish allergy among older children in the Philippines, Singapore and Thailand. Methods: The population prevalence of fish allergy in 14- to 16-year-old children in the 3 countries was evaluated using a structured written questionnaire which was distributed to students of randomly selected secondary schools. An extended questionnaire to determine convincing fish allergy on the basis of typical clinical manifestations within 2 h of ingestion was administered to those with positive responses. Results: From acohort of 25,842 students, responses were 81.1% in the Philippines (n = 11,434), 67.9% in Singapore (n = 6,498) and 80.2% (n = 2,034) in Thailand. Using criteria for convincing food allergy, fish allergy was much higher in the Philippines [2.29%, 95% confidence interval (CI) 2.02–2.56] than in Singapore (0.26%, 95% CI 0.14–0.79) and Thailand (0.29%, 95% CI 0.06–0.52). Weighted multiple logistic regression analyses showed that compared to the Philippines, prevalence rates were lower in Singapore [odds ratio (OR) 0.40, 95% CI 0.27–0.60, p < 0.0001] and Thailand (OR 0.13, 95% CI 0.05–0.33, p < 0.0001). Females were more likely to have fish allergy compared to males for all children combined (OR 1.32, 95% CI 1.11–1.58, p = 0.002). Most allergies appeared mild, as only 28% of cases sought medical consultation at the time of the reaction and 31.2% of cases reported continued exposure despite allergic symptoms. Conclusion: Fish allergy in late childhood is more common in the Philippines compared to Singapore and Thailand. Differences in food processing, dietary habits and other cultural practices might be important risk factors for the development of fish allergy in these populations.
Enhancement of the Frequency and Function of IL-10-Secreting Type I T Regulatory Cells after 1 Year of Cluster Allergen-Specific Immunotherapy
Authors: Lou Wei , Wang Chengshuo , Wang Yang , Han Demin , Zhang Luo
Published: July 2012

Background: Allergen-specific immunotherapy (SIT) is a highly effective treatment for allergic diseases, but the underlying mechanisms are unclear. In this study, we investigated the effects of cluster SIT with Dermatophagoides pteronyssinus (Der p) on IL-10-secreting type I T regulatory (Tr1) cells in Der p-sensitized patients with allergic rhinitis. Methods: This was a prospective randomized study involving 68 participants (aged 18–60 years) of whom 38 were patients with allergic rhinitis and received Der p-SIT for 1 year and 30 were nonallergic controls. IL-10+IL-4–CD4+ T cells were measured by flow cytometry for the patient group at baseline and at the end of 1 year of SIT, and for nonallergic controls. Similarly, IL-10 in supernatants from allergen-stimulated peripheral blood mononuclear cell (PBMC) cultures were measured by ELISA, and the suppressive effect of Tr1 cells on cell proliferation and cytokine release (IFN-γ and IL-4) in PBMCs was estimated in cultures from both groups. Allergen-specific serum IgE and IgG4 were also assessed by RAST and ELISA for the SIT group. Results: The levels of IL-10-secreting Tr1 cells, IgG4 and allergen-induced IL-10 synthesis from PBMC cultures were significantly increased and the function of Tr1 cells was enhanced after 1 year of SIT compared to baseline levels. In contrast, the level of IgE was not significantly changed. Conclusion: These data suggest that the cluster Der p-SIT may enhance the frequency and function of IL-10-secreting Tr1 cells.
Nizatidine Improves Clinical Symptoms and Gastric Emptying in Patients with Functional Dyspepsia Accompanied by Impaired Gastric Emptying
Authors: Futagami S. , Shimpuku M. , Song J.M. , Kodaka Y. , Yamawaki H. , Nagoya H. , Shindo T. , Kawagoe T. , Horie A. , Gudis K. , Iwakiri K. , Sakamoto C.
Published: July 2012

Background/Aims: In this crossover study, we investigated whether nizatidine, a H2-receptor antagonist, can alleviate clinical symptoms and gastric emptying in patients with Rome III-based functional dyspepsia (FD) with or without impaired gastric emptying. Methods: We enrolled 30 patients presenting with FD symptoms (epigastric pain syndrome, n = 6; postprandial distress syndrome, n = 24). Rome III-based FD patients were treated with nizatidine (300 mg/day) or placebo for 4 weeks in a crossover trial. Gastric motility was mainly evaluated with the Tmax value using the 13C-acetate breath test. Meal-related symptoms were defined as postprandial fullness and early satiation. Gastroesophageal symptom was defined as a burning feeling rising from the stomach or lower chest up toward the neck. Acylated- and desacylated ghrelin levels were evaluated by the ELISA method. Clinical symptoms, gastric emptying and ghrelin levels were evaluated at three different points during the study (pretreatment, after 4 weeks former treatment and after 4 weeks later treatment). The primary end point of this study was to determine whether nizatidine would improve clinical symptoms and gastric emptying in FD patients with or without impaired gastric emptying via affecting ghrelin levels. Results: Meal-related symptoms of the patients treated with nizatidine improved significantly (21/30; 70%) compared to those treated with placebo (3/30; 10%). In addition, nizatidine treatment also significantly improved gastroesophageal symptoms (16/30; 53%) compared to those treated with placebo (0/30; 0%). Nizatidine treatment in patients with FD accompanied by impaired gastric emptying significantly improved clinical symptoms and Tmax value as a marker of gastric emptying (10/11, 91%; 9/11, 82%) compared to placebo therapy, respectively. There were no significant differences in ghrelin levels between nizatidine treatment and placebo therapy. Conclusion: Nizatidine administration significantly improved both gastric emptying and clinical symptoms in FD patients with impaired gastric emptying.