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http://www.ncbi.nlm.nih.gov/pubmed/23096376
1. G Ital Cardiol (Rome). 2012 Oct;13(10 Suppl 2):50S-54S. doi: 10.1714/1167.12921. [New pharmacological approaches to ischemic heart disease]. [Article in Italian] Raddino R(1), Della Pina P, Gorga E, Brambilla G, Regazzoni V, Gavazzoni M, Dei Cas L. Author information: (1)Dipartimento di Medicina Sperimentale ed Applicata, Universita degli Studi e Spedali Civili, Brescia. riccardo.raddino@libero.it Major steps have been made in the treatment of ischemic heart disease from the discovery of nitrates as antianginal medication to the techniques of percutaneous angioplasty. This incredible therapeutic progress has resulted in a reduced incidence of ischemic heart disease and related mortality and morbidity. However, statistical and epidemiological data indicate that in ischemic heart disease, despite the achievement of great success, there is a necessity for a further step toward treatment, considering the fact that the characteristics of this population are changing (increased prevalence of subendocardial infarction compared with classic transmural infarction, especially in the elderly population). Furthermore, the need for alternative therapeutic approaches to traditional ones is recognized. Ranolazine is a selective inhibitor of Na channels that prevents pathological extension of late Na current developing in the ischemic myocardial cell. This current is responsible for calcium overload, with consequent impairment of diastolic relaxation. Ranolazine reduces Na overload induced by calcium and improves diastolic relaxation and coronary subendocardial flow, without affecting hemodynamic parameters such as blood pressure, heart rate, or inotropic state of the heart, avoiding undesirable side effects. Efficacy of ranolazine has been evaluated in several trials, using clinical and instrumental endpoints (MARISA and CARISA) or, more recently, using endpoints such as mortality and reinfarction (ERICA and MERLIN-TIMI 36). Ivabradine acts through the inhibition of late Na current (also known as If), which controls the spontaneous diastolic depolarization of sinus node cells. The partial inhibition of these channels reduces the frequency of sinus node action potential initiation, resulting in decreased heart rate without effects on contractility, atrio-ventricular conduction, or repolarization. The BEAUTIFUL trial has tested whether the effect of ivabradine in lowering heart rate is able to reduce mortality and cardiovascular morbidity in patients with coronary artery disease and left ventricular systolic dysfunction. The most significant results were obtained in the subgroup of patients with life-limiting exertional angina. In this group, ivabradine significantly reduced the primary endpoint, a composite of cardiovascular death, hospitalization for fatal and nonfatal acute myocardial infarction (AMI) or heart failure, by 24%, and hospitalizations for AMI by 42%. In the subgroup of patients with baseline heart rate >70 bpm, hospitalizations for AMI and revascularization were reduced by 73% and 59%, respectively. DOI: 10.1714/1167.12921 PMID: 23096376 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/14722917
1. Hum Mutat. 2004 Feb;23(2):134-146. doi: 10.1002/humu.10305. Characterization of the somatic mutational spectrum of the neurofibromatosis type 1 (NF1) gene in neurofibromatosis patients with benign and malignant tumors. Upadhyaya M(1), Han S(1), Consoli C(1), Majounie E(1), Horan M(1), Thomas NS(1), Potts C(1), Griffiths S(1), Ruggieri M(2), von Deimling A(3), Cooper DN(1). Author information: (1)Institute of Medical Genetics, University of Wales College of Medicine, Cardiff, UK. (2)Institute of Bioimaging and Pathology of the Central Nervous System, National Research Council, Catania, Italy. (3)Institut für Neuropathologie, Charité, Humboldt-Universität, Berlin, Germany. One of the main features of neurofibromatosis type 1 (NF1) is benign neurofibromas, 10-20% of which become transformed into malignant peripheral nerve sheath tumors (MPNSTs). The molecular basis of NF1 tumorigenesis is, however, still unclear. Ninety-one tumors from 31 NF1 patients were screened for gross changes in the NF1 gene using microsatellite/restriction fragment length polymorphism (RFLP) markers; loss of heterozygosity (LOH) was found in 17 out of 91 (19%) tumors (including two out of seven MPNSTs). Denaturing high performance liquid chromatography (DHPLC) was then used to screen 43 LOH-negative and 10 LOH-positive tumors for NF1 microlesions at both RNA and DNA levels. Thirteen germline and 12 somatic mutations were identified, of which three germline (IVS7-2A>G, 3731delT, 6117delG) and eight somatic (1888delG, 4374-4375delCC, R2129S, 2088delG, 2341del18, IVS27b-5C>T, 4083insT, Q519P) were novel. A mosaic mutation (R2429X) was also identified in a neurofibroma by DHPLC analysis and cloning/sequencing. The observed somatic and germline mutational spectra were similar in terms of mutation type, relative frequency of occurrence, and putative underlying mechanisms of mutagenesis. Tumors lacking mutations were screened for NF1 gene promoter hypermethylation but none were found. Microsatellite instability (MSI) analysis revealed MSI in five out of 11 MPNSTs as compared to none out of 70 neurofibromas (p=1.8 x 10(-5)). The screening of seven MPNSTs for subtle mutations in the CDKN2A and TP53 genes proved negative, although the screening of 11 MPNSTs detected LOH involving either the TP53 or the CDKN2A gene in a total of four tumors. These findings are consistent with the view that NF1 tumorigenesis is a complex multistep process involving a variety of different types of genetic defect at multiple loci. Copyright 2003 Wiley-Liss, Inc. DOI: 10.1002/humu.10305 PMID: 14722917 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16158195
1. Biosci Rep. 2004 Dec;24(6):559-76. doi: 10.1007/s10540-005-2792-x. Pre-UV-treatment of cells results in enhanced host cell reactivation of a UV damaged reporter gene in CHO-AA8 chinese hamster ovary cells but not in transcription-coupled repair deficient CHO-UV61 cells. Liu L(1), Rainbow AJ. Author information: (1)Department of Biology, McMaster University, L8S 4K1, Hamilton, Ontario, Canada, rainbow@mcmaster.ca. We have used a non-replicating recombinant adenovirus, Ad5MCMVlacZ, which expresses the beta-galactosidase reporter gene, to examine both constitutive and inducible repair of UV-damaged DNA in repair proficient CHO-AA8 Chinese hamster ovary cells and in mutant CHO-UV61 cells which are deficient in the transcription-coupled repair (TCR) pathway of nucleotide excision repair. Host cell reactivation (HCR) of beta-galactosidase activity for UV-irradiated Ad5MCMVlacZ was significantly reduced in non-irradiated CHO-UV61 cells compared to that in non-irradiated CHO-AA8 cells suggesting that repair in the transcribed strand of the UV-damaged reporter gene in untreated cells utilizes TCR. Prior UV-irradiation of cells with low UV fluences resulted in a transient enhancement of HCR for expression of the UV-damaged reporter gene in CHO-AA8 cells but not in TCR deficient CHO-UV61 cells. These results suggest the presence of an inducible DNA pathway in CHO cells that results from an enhancement of TCR or a mechanism that involves the TCR pathway. DOI: 10.1007/s10540-005-2792-x PMID: 16158195 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12142466
1. Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10571-4. doi: 10.1073/pnas.162278199. Epub 2002 Jul 25. Clusters of transcription-coupled repair in the human genome. Surrallés J(1), Ramírez MJ, Marcos R, Natarajan AT, Mullenders LH. Author information: (1)Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain. jordi.surralles@uab.es A specialized nucleotide excision repair pathway known as transcription-coupled repair (TCR) counteracts the toxic effects of DNA damage in transcriptionally active genes. The clustering of active genes into gene-rich chromosomal domains predicts that the sites of TCR are unevenly distributed through the genome. To elucidate the genomic organization and chromosomal localization of TCR, we isolated DNA fragments encompassing TCR-mediated repair sites from UV-C irradiated xeroderma pigmentosum group C cells, which can only repair the transcribed strand of active genes. This DNA was used as a molecular probe to visualize TCR in normal metaphase spreads by reverse fluorescence in situ hybridization. Whereas DNA repair sites in normal human cells are evenly distributed through the genome, TCR is highly localized at specific chromosomal domains. Particularly, clusters of TCR sites were identified at early-replicating gene-rich bands and telomeric regions of several chromosomes. High gene-density chromosomes such as chromosome 19 and the GC-rich domains of several chromosomes (T bands) are preferential locations of TCR. Our results demonstrate that the intragenomic localization of TCR resembles the uneven distribution of the human transcriptome, CpG islands, and hyperacetylated histones, enforcing the basic link between DNA repair, transcription, and nuclear organization in a complex genome. DOI: 10.1073/pnas.162278199 PMCID: PMC124978 PMID: 12142466 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/7957102
1. EMBO J. 1994 Nov 15;13(22):5361-9. doi: 10.1002/j.1460-2075.1994.tb06871.x. RAD26, the functional S. cerevisiae homolog of the Cockayne syndrome B gene ERCC6. van Gool AJ(1), Verhage R, Swagemakers SM, van de Putte P, Brouwer J, Troelstra C, Bootsma D, Hoeijmakers JH. Author information: (1)MGC Department of Cell Biology and Genetics, Erasmus University Rotterdam, The Netherlands. Transcription-coupled repair (TCR) is a universal sub-pathway of the nucleotide excision repair (NER) system that is limited to the transcribed strand of active structural genes. It accomplishes the preferential elimination of transcription-blocking DNA lesions and permits rapid resumption of the vital process of transcription. A defect in TCR is responsible for the rare hereditary disorder Cockayne syndrome (CS). Recently we found that mutations in the ERCC6 repair gene, encoding a putative helicase, underly the repair defect of CS complementation group B. Here we report the cloning and characterization of the Saccharomyces cerevisiae homolog of CSB/ERCC6, which we designate RAD26. A rad26 disruption mutant appears viable and grows normally, indicating that the gene does not have an essential function. In analogy with CS, preferential repair of UV-induced cyclobutane pyrimidine dimers in the transcribed strand of the active RBP2 gene is severely impaired. Surprisingly, in contrast to the human CS mutant, yeast RAD26 disruption does not induce any UV-, cisPt- or X-ray sensitivity, explaining why it was not isolated as a mutant before. Recovery of growth after UV exposure was somewhat delayed in rad26. These findings suggest that TCR in lower eukaryotes is not very important for cell survival and that the global genome repair pathway of NER is the major determinant of cellular resistance to genotoxicity. DOI: 10.1002/j.1460-2075.1994.tb06871.x PMCID: PMC395493 PMID: 7957102 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24920614
1. J Neurosci. 2014 Jun 11;34(24):8083-97. doi: 10.1523/JNEUROSCI.0543-14.2014. Profilin 1 associates with stress granules and ALS-linked mutations alter stress granule dynamics. Figley MD(1), Bieri G(1), Kolaitis RM(2), Taylor JP(2), Gitler AD(3). Author information: (1)Department of Genetics and Neuroscience Graduate Program, Stanford University School of Medicine, Stanford, California 94305, and. (2)Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105. (3)Department of Genetics and agitler@stanford.edu. Mutations in the PFN1 gene encoding profilin 1 are a rare cause of familial amyotrophic lateral sclerosis (ALS). Profilin 1 is a well studied actin-binding protein but how PFN1 mutations cause ALS is unknown. The budding yeast, Saccharomyces cerevisiae, has one PFN1 ortholog. We expressed the ALS-linked profilin 1 mutant proteins in yeast, demonstrating a loss of protein stability and failure to restore growth to profilin mutant cells, without exhibiting gain-of-function toxicity. This model provides for simple and rapid screening of novel ALS-linked PFN1 variants. To gain insight into potential novel roles for profilin 1, we performed an unbiased, genome-wide synthetic lethal screen with yeast cells lacking profilin (pfy1Δ). Unexpectedly, deletion of several stress granule and processing body genes, including pbp1Δ, were found to be synthetic lethal with pfy1Δ. Mutations in ATXN2, the human ortholog of PBP1, are a known ALS genetic risk factor and ataxin 2 is a stress granule component in mammalian cells. Given this genetic interaction and recent evidence linking stress granule dynamics to ALS pathogenesis, we hypothesized that profilin 1 might also associate with stress granules. Here we report that profilin 1 and related protein profilin 2 are novel stress granule-associated proteins in mouse primary cortical neurons and in human cell lines and that ALS-linked mutations in profilin 1 alter stress granule dynamics, providing further evidence for the potential role of stress granules in ALS pathogenesis. Copyright © 2014 the authors 0270-6474/14/348083-15$15.00/0. DOI: 10.1523/JNEUROSCI.0543-14.2014 PMCID: PMC4051967 PMID: 24920614 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24090136
1. Mol Neurodegener. 2013 Aug 31;8:30. doi: 10.1186/1750-1326-8-30. Amyotrophic lateral sclerosis-linked FUS/TLS alters stress granule assembly and dynamics. Baron DM(1), Kaushansky LJ, Ward CL, Sama RR, Chian RJ, Boggio KJ, Quaresma AJ, Nickerson JA, Bosco DA. Author information: (1)Department of Neurology, University of Massachusetts Medical School, Worcester, MA, USA. Daryl.Bosco@umassmed.edu. BACKGROUND: Amyotrophic lateral sclerosis (ALS)-linked fused in sarcoma/translocated in liposarcoma (FUS/TLS or FUS) is concentrated within cytoplasmic stress granules under conditions of induced stress. Since only the mutants, but not the endogenous wild-type FUS, are associated with stress granules under most of the stress conditions reported to date, the relationship between FUS and stress granules represents a mutant-specific phenotype and thus may be of significance in mutant-induced pathogenesis. While the association of mutant-FUS with stress granules is well established, the effect of the mutant protein on stress granules has not been examined. Here we investigated the effect of mutant-FUS on stress granule formation and dynamics under conditions of oxidative stress. RESULTS: We found that expression of mutant-FUS delays the assembly of stress granules. However, once stress granules containing mutant-FUS are formed, they are more dynamic, larger and more abundant compared to stress granules lacking FUS. Once stress is removed, stress granules disassemble more rapidly in cells expressing mutant-FUS. These effects directly correlate with the degree of mutant-FUS cytoplasmic localization, which is induced by mutations in the nuclear localization signal of the protein. We also determine that the RGG domains within FUS play a key role in its association to stress granules. While there has been speculation that arginine methylation within these RGG domains modulates the incorporation of FUS into stress granules, our results demonstrate that this post-translational modification is not involved. CONCLUSIONS: Our results indicate that mutant-FUS alters the dynamic properties of stress granules, which is consistent with a gain-of-toxic mechanism for mutant-FUS in stress granule assembly and cellular stress response. DOI: 10.1186/1750-1326-8-30 PMCID: PMC3766239 PMID: 24090136 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/1832527
1. Am Rev Respir Dis. 1991 Sep;144(3 Pt 1):564-9. doi: 10.1164/ajrccm/144.3_Pt_1.564. Activity of clarithromycin against Mycobacterium avium infection in patients with the acquired immune deficiency syndrome. A controlled clinical trial. Dautzenberg B(1), Truffot C, Legris S, Meyohas MC, Berlie HC, Mercat A, Chevret S, Grosset J. Author information: (1)Pulmonary Department Groupe Hospitalier Pitié-Salpêtrière, Paris, France. Comment in Am Rev Respir Dis. 1992 May;145(5):1241-2. doi: 10.1164/ajrccm/145.5.1241b. Disseminated Mycobacterium avium infection is common in patients with acquired immune deficiency syndrome (AIDS), but no drug studies have been reported establishing antimicrobial activity against this organism in a controlled, randomized trial. Clarithromycin, a new macrolide, has activity against M. avium in vitro and in animals, but it has not been studied in humans. In this randomized, double-blind, placebo-controlled trial, we measured the ability of clarithromycin to reduce M. avium bacteremia in patients with AIDS and disseminated infection. Of 23 patients initially enrolled, 15 men with late-stage AIDS qualified for the study. One group received clarithromycin alone for 6 wk, then placebo plus rifampin, isoniazid, ethambutol, and clofazimine for 6 wk. The other group received placebo alone, then clarithromycin plus the other four drugs. Colony-forming units (CFU) of M. avium per milliliter of blood were determined by quantitative cultures taken at baseline and every 2 wk thereafter. Minimum inhibitory concentration of clarithromycin for 90% of the strains isolated from patients at baseline, as measured on 7H11 agar at pH 6.6, was 8 micrograms/ml. Eight eligible patients with initial positive cultures who were initially receiving clarithromycin alone had marked declines in blood M. avium CFU; in six cases, CFU decreased to zero. When seven patients were switched to placebo plus the other four drugs, CFU rose in four patients and remained undetectable in three. The five eligible patients initially treated with placebo had progressive CFU increases; when three were switched to clarithromycin plus the four drugs, their CFU declined.(ABSTRACT TRUNCATED AT 250 WORDS) DOI: 10.1164/ajrccm/144.3_Pt_1.564 PMID: 1832527 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11073759
1. Clin Infect Dis. 2000 Nov;31(5):1245-52. doi: 10.1086/317468. Epub 2000 Nov 6. A randomized, double-blind trial comparing azithromycin and clarithromycin in the treatment of disseminated Mycobacterium avium infection in patients with human immunodeficiency virus. Dunne M(1), Fessel J, Kumar P, Dickenson G, Keiser P, Boulos M, Mogyros M, White AC Jr, Cahn P, O'Connor M, Lewi D, Green S, Tilles J, Hicks C, Bissett J, Schneider MM, Benner R. Author information: (1)Pfizer Central Research, Groton, CT 06340, USA. dunnemw@groton.pfizer.com Erratum in Clin Infect Dis 2001 May 1;32(9):1386. Two hundred and forty-six patients infected with human immunodeficiency virus (HIV) who also had disseminated Mycobacterium avium complex received either azithromycin 250 mg every day, azithromycin 600 mg every day, or clarithromycin 500 mg twice a day, each combined with ethambutol, for 24 weeks. Samples drawn from patients were cultured and clinically assessed every 3 weeks up to week 12, then monthly thereafter through week 24 of double-blind therapy and every 3 months while on open-label therapy through the conclusion of the trial. The azithromycin 250 mg arm of the study was dropped after an interim analysis showed a lower rate of clearance of bacteremia. At 24 weeks of therapy, the likelihood of patients' developing 2 consecutive negative cultures (46% vs. 56%, P=.24) or 1 negative culture (59% vs. 61%, P=.80) was similar for azithromycin 600 mg (n=68) and clarithromycin (n=57), respectively. The likelihood of relapse was 39% versus 27% (P=.21) on azithromycin compared with clarithromycin, respectively. Of the 6 patients who experienced relapse, none of those randomized to receive azithromycin developed isolates resistant to macrolides, compared with 2 of 3 patients randomized to receive clarithromycin [corrected]. Mortality was similar in patients comprising each arm of the study (69% vs. 63%; hazard, 95.1% confidence interval, 1.1 [0.7, 1.7]). Azithromycin 600 mg, when given in combination with ethambutol, is an effective agent for the treatment of disseminated M. avium disease in patients infected with HIV. DOI: 10.1086/317468 PMID: 11073759 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/9875582
1. AIDS. 1998 Dec 24;12(18):2439-46. doi: 10.1097/00002030-199818000-00013. A phase II/III trial of antimicrobial therapy with or without amikacin in the treatment of disseminated Mycobacterium avium infection in HIV-infected individuals. AIDS Clinical Trials Group Protocol 135 Study Team. Parenti DM(1), Williams PL, Hafner R, Jacobs MR, Hojczyk P, Hooton TM, Barber TW, Simpson G, van der Horst C, Currier J, Powderly WG, Limjoco M, Ellner JJ. Author information: (1)George Washington University Medical Center, Washington, DC 20037, USA. OBJECTIVE: To determine the clinical and microbiologic benefit of adding amikacin to a four-drug oral regimen for treatment of disseminated Mycobacterium avium infection in HIV-infected patients. DESIGN: A randomized, open-labeled, comparative trial. SETTING: Outpatient clinics. PATIENTS: Seventy-four patients with HIV and symptomatic bacteremic M. avium infection. INTERVENTIONS: Rifampin 10 mg/kg daily, ciprofloxacin 500 mg twice daily, clofazimine 100 mg every day, and ethambutol 15 mg/kg orally daily for 24 weeks, with or without amikacin 10 mg/kg intravenously or intramuscularly 5 days weekly for the first 4 weeks. MAIN OUTCOME MEASURE: Clinical and microbiologic response at 4 weeks; quantitative level of bacteremia with M. avium. RESULTS: No difference in clinical response was noted with the addition of amikacin to the four-drug oral regimen, and only 25% in either group had a complete or partial response at 4 weeks. A comparable quantitative decrease in bacteremia was noted in both treatment groups, with 16% of patients being culture-negative at 4 weeks and 38% at 12 weeks. Toxicities were mainly gastrointestinal. Amikacin was well tolerated. Median survival was 30 weeks in both groups. CONCLUSIONS: The addition of amikacin to a four-drug oral regimen of rifampin, ciprofloxacin, clofazimine, and ethambutol did not provide clinical or microbiologic benefit. DOI: 10.1097/00002030-199818000-00013 PMID: 9875582 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23152885
1. PLoS One. 2012;7(11):e49267. doi: 10.1371/journal.pone.0049267. Epub 2012 Nov 13. The effect of PRMT1-mediated arginine methylation on the subcellular localization, stress granules, and detergent-insoluble aggregates of FUS/TLS. Yamaguchi A(1), Kitajo K. Author information: (1)Department of Neurobiology, Graduate School of Medicine, Chiba University, Chiba, Japan. Fused in sarcoma/translocated in liposarcoma (FUS/TLS) is one of causative genes for familial amyotrophic lateral sclerosis (ALS). In order to identify binding partners for FUS/TLS, we performed a yeast two-hybrid screening and found that protein arginine methyltransferase 1 (PRMT1) is one of binding partners primarily in the nucleus. In vitro and in vivo methylation assays showed that FUS/TLS could be methylated by PRMT1. The modulation of arginine methylation levels by a general methyltransferase inhibitor or conditional over-expression of PRMT1 altered slightly the nucleus-cytoplasmic ratio of FUS/TLS in cell fractionation assays. Although co-localized primarily in the nucleus in normal condition, FUS/TLS and PRMT1 were partially recruited to the cytoplasmic granules under oxidative stress, which were merged with stress granules (SGs) markers in SH-SY5Y cell. C-terminal truncated form of FUS/TLS (FUS-dC), which lacks C-terminal nuclear localization signal (NLS), formed cytoplasmic inclusions like ALS-linked FUS mutants and was partially co-localized with PRMT1. Furthermore, conditional over-expression of PRMT1 reduced the FUS-dC-mediated SGs formation and the detergent-insoluble aggregates in HEK293 cells. These findings indicate that PRMT1-mediated arginine methylation could be implicated in the nucleus-cytoplasmic shuttling of FUS/TLS and in the SGs formation and the detergent-insoluble inclusions of ALS-linked FUS/TLS mutants. DOI: 10.1371/journal.pone.0049267 PMCID: PMC3496700 PMID: 23152885 [Indexed for MEDLINE] Conflict of interest statement: Competing Interests: The authors have declared that no competing interests exist.
http://www.ncbi.nlm.nih.gov/pubmed/17431895
1. Am J Med Genet A. 2007 May 1;143A(9):999-1008. doi: 10.1002/ajmg.a.31689. Smith-Magenis syndrome and Moyamoya disease in a patient with del(17)(p11.2p13.1). Girirajan S(1), Mendoza-Londono R, Vlangos CN, Dupuis L, Nowak NJ, Bunyan DJ, Hatchwell E, Elsea SH. Author information: (1)Department of Human Genetics, Medical College of Virginia Campus, Virginia Commonwealth University, Richmond, VA 23298, USA. Chromosomal rearrangements causing microdeletions and microduplications are a major cause of congenital malformation and mental retardation. Because they are not visible by routine chromosome analysis, high resolution whole-genome technologies are required for the detection and diagnosis of small chromosomal abnormalities. Recently, array-comparative genomic hybridization (aCGH) and multiplex ligation-dependent probe amplification (MLPA) have been useful tools for the identification and mapping of deletions and duplications at higher resolution and throughput. Smith-Magenis syndrome (SMS) is a multiple congenital anomalies/mental retardation syndrome caused by deletion or mutation of the retinoic acid induced 1 (RAI1) gene and is often associated with a chromosome 17p11.2 deletion. We report here on the clinical and molecular analysis of a 10-year-old girl with SMS and moyamoya disease (occlusion of the circle of Willis). We have employed a combination of aCGH, FISH, and MLPA to characterize an approximately 6.3 Mb deletion spanning chromosome region 17p11.2-p13.1 in this patient, with the proximal breakpoint within the RAI1 gene. Further, investigation of the genomic architecture at the breakpoint intervals of this large deletion documented the presence of palindromic repeat elements that could potentially form recombination substrates leading to unequal crossover. DOI: 10.1002/ajmg.a.31689 PMID: 17431895 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23121133
1. Dev Med Child Neurol. 2013 Feb;55(2):167-172. doi: 10.1111/dmcn.12013. Epub 2012 Nov 2. Brain magnetic resonance imaging and motor and intellectual functioning in 86 patients born at term with spastic diplegia. Numata Y(1)(2), Onuma A(1), Kobayashi Y(3), Sato-Shirai I(1)(2), Tanaka S(1), Kobayashi S(1), Wakusawa K(1), Inui T(1), Kure S(2), Haginoya K(1). Author information: (1)Department of Pediatric Neurology, Takuto Rehabilitation Center for Children, Sendai. (2)Department of Pediatrics, Tohoku University School of Medicine, Sendai. (3)Department of Pediatrics, Nishitaga National Hospital, Sendai, Japan. Comment in Dev Med Child Neurol. 2013 Feb;55(2):103-104. doi: 10.1111/dmcn.12047. AIM: To investigate the association between magnetic resonance imaging (MRI) patterns and motor function, epileptic episodes, and IQ or developmental quotient in patients born at term with spastic diplegia. METHOD: Eighty-six patients born at term with cerebral palsy (CP) and spastic diplegia (54 males, 32 females; median age 20 y, range 7-42 y) among 829 patients with CP underwent brain MRI between 1990 and 2008. The MRI and clinical findings were analysed retrospectively. Intellectual disability was classified according to the Enjoji developmental test or the Wechsler Intelligence Scale for Children (3rd edition). RESULTS: The median ages at diagnosis of CP, assignment of Gross Motor Function Classification System (GMFCS) level, cognitive assessment, and MRI were 2 years (range 5 mo-8 y), 6 years (2 y 8 mo-19 y), 6 years (1 y 4 mo-19 y), and 7 years (10 mo-30 y) respectively. MRI included normal findings (41.9%), periventricular leukomalacia, hypomyelination, and porencephaly/periventricular venous infarction. The frequency of patients in GMFCS levels III to V and intellectual disability did not differ between those with normal and abnormal MRI findings. Patients with normal MRI findings had significantly fewer epileptic episodes than those with abnormal ones (p=0.001). INTERPRETATION: Varied MRI findings, as well as the presence of severe motor dysfunction and intellectual disability (despite normal MRI), suggest that patients born at term with spastic diplegia had heterogeneous and unidentified pathophysiology. © The Authors. Developmental Medicine & Child Neurology © 2012 Mac Keith Press. DOI: 10.1111/dmcn.12013 PMID: 23121133 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19765185
1. J Neurochem. 2009 Nov;111(4):1051-61. doi: 10.1111/j.1471-4159.2009.06383.x. Epub 2009 Sep 16. TDP-43 is recruited to stress granules in conditions of oxidative insult. Colombrita C(1), Zennaro E, Fallini C, Weber M, Sommacal A, Buratti E, Silani V, Ratti A. Author information: (1)Department of Neurology and Laboratory of Neuroscience, Dino Ferrari Center, Università degli Studi di Milano - IRCCS Istituto Auxologico Italiano, Milan, Italy. Transactive response DNA-binding protein 43 (TDP-43) forms abnormal ubiquitinated and phosphorylated inclusions in brain tissues from patients with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. TDP-43 is a DNA/RNA-binding protein involved in RNA processing, such as transcription, pre-mRNA splicing, mRNA stabilization and transport to dendrites. We found that in response to oxidative stress and to environmental insults of different types TDP-43 is capable to assemble into stress granules (SGs), ribonucleoprotein complexes where protein synthesis is temporarily arrested. We demonstrated that a specific aminoacidic interval (216-315) in the C-terminal region and the RNA-recognition motif 1 domain are both implicated in TDP-43 participation in SGs as their deletion prevented the recruitment of TDP-43 into SGs. Our data show that TDP-43 is a specific component of SGs and not of processing bodies, although we proved that TDP-43 is not necessary for SG formation, and its gene silencing does not impair cell survival during stress. The analysis of spinal cord tissue from ALS patients showed that SG markers are not entrapped in TDP-43 pathological inclusions. Although SGs were not evident in ALS brains, we speculate that an altered control of mRNA translation in stressful conditions may trigger motor neuron degeneration at early stages of the disease. DOI: 10.1111/j.1471-4159.2009.06383.x PMID: 19765185 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15563017
1. J Child Neurol. 2004 Sep;19(9):699-709. doi: 10.1177/08830738040190091101. Molecular genetic basis of tuberous sclerosis complex: from bench to bedside. Au KS(1), Williams AT, Gambello MJ, Northrup H. Author information: (1)Department of Pediatrics, Division of Medical Genetics, The University of Texas Medical School at Houston, Houston, TX 77030, USA. Tuberous sclerosis complex is an autosomal dominant disease of benign tumors occurring in multiple organ systems of the body. Either of two genes, TSC1 or TSC2, can be mutated, resulting in the tuberous sclerosis complex phenotype. The protein products of the tuberous sclerosis complex genes, hamartin (TSC1) and tuberin (TSC2), have been discovered to play important roles in several cell-signaling pathways. Knowledge regarding the function of the tuberin-hamartin complex has led to therapeutic intervention trials. Numerous pathogenic mutations have been elucidated in individuals affected with tuberous sclerosis complex. Information on the type and distribution of nearly 1000 mutations in the two genes is discussed. Mosaicism for tuberous sclerosis complex mutations has been documented, complicating provision of genetic counseling to families. Emerging genotype-phenotype correlations should provide guidance for better medical care of individuals with tuberous sclerosis complex. DOI: 10.1177/08830738040190091101 PMID: 15563017 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21177652
1. Nucleic Acids Res. 2011 Apr;39(8):3103-15. doi: 10.1093/nar/gkq1298. Epub 2010 Dec 21. The chromodomains of CHD1 are critical for enzymatic activity but less important for chromatin localization. Morettini S(1), Tribus M, Zeilner A, Sebald J, Campo-Fernandez B, Scheran G, Wörle H, Podhraski V, Fyodorov DV, Lusser A. Author information: (1)Division of Molecular Biology, Biocenter, Innsbruck Medical University, Fritz-Pregl Strasse 3, 6020 Innsbruck, Austria. The molecular motor protein CHD1 has been implicated in the regulation of transcription and in the transcription-independent genome-wide incorporation of H3.3 into paternal chromatin in Drosophila melanogaster. A key feature of CHD1 is the presence of two chromodomains, which can bind to histone H3 methylated at lysine 4 and thus might serve to recruit and/or maintain CHD1 at the chromatin. Here, we describe genetic and biochemical approaches to the study of the Drosophila CHD1 chromodomains. We found that overall localization of CHD1 on polytene chromosomes does not appreciably change in chromodomain-mutant flies. In contrast, the chromodomains are important for transcription-independent activities of CHD1 during early embryonic development as well as for transcriptional regulation of several heat shock genes. However, neither CHD1 nor its chromodomains are needed for RNA polymerase II localization and H3K4 methylation but loss of CHD1 decreases transcription-induced histone eviction at the Hsp70 gene in vivo. Chromodomain mutations negatively affect the chromatin assembly activities of CHD1 in vitro, and they appear to be involved in linking the ATP-dependent motor to the chromatin assembly function of CHD1. DOI: 10.1093/nar/gkq1298 PMCID: PMC3082874 PMID: 21177652 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16468993
1. Mol Microbiol. 2006 Mar;59(5):1531-41. doi: 10.1111/j.1365-2958.2005.05031.x. The ISWI and CHD1 chromatin remodelling activities influence ADH2 expression and chromatin organization. Xella B(1), Goding C, Agricola E, Di Mauro E, Caserta M. Author information: (1)Istituto Pasteur-Fondazione Cenci Bolognetti, c/o Dipartmento di Genetica e Biologia Molecolare, Universita La Sapienza, 00185 Rome, Italy. Nucleosome remodelling complexes play a key role in gene activation in response to environmental changes by driving promoter chromatin to reach an accessible configuration. They also mediate genome-wide chromatin organization, although their role in processes other than activation-related chromatin remodelling are poorly understood. The Saccharomyces cerevisiae ADH2 gene represents an excellent model for understanding the role of chromatin structure and remodelling in gene regulation. Following glucose depletion, highly positioned promoter nucleosomes are destabilized leading to strictly regulated kinetics of transcriptional activation. Nevertheless, no chromatin remodelling activities responsible for establishing or remodelling ADH2 chromatin structure have been identified to date. Here we show that the absence of the Isw1 and Chd1 ATP-dependent chromatin remodelling activities delays the maximal expression of ADH2 without impairing the chromatin remodelling that occurs upon activation. Instead, a destabilized chromatin structure on the ADH2 coding and termination region is observed in the absence of Isw1 or Chd1 in repressing conditions. The specific Isw1 complex involved in this nucleosome repositioning is Isw1b because the deletion of Ioc2 and Ioc4, but not of Ioc3, causes the same phenotype as the deletion of Isw1. Moreover, the lack of Chd1 combined with the absence of Isw1 and Isw2 impairs nucleosome spacing along the ADH2 gene, and genome-wide in S. cerevisiae. Thus, the ISWI and Chd1 remodelling factors are not only involved in transcription-related chromatin remodelling, but also are required to maintain a specific chromatin configuration across the yeast genome. DOI: 10.1111/j.1365-2958.2005.05031.x PMID: 16468993 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18576213
1. Int J Neurosci. 2008 Aug;118(8):1157-63. doi: 10.1080/00207450801898279. Neurofibromatosis type 1 association with moyamoya disease. Koc F(1), Yerdelen D, Koc Z. Author information: (1)Department of Neurology, Cukurova University Medical School, Adana, Turkey. kocfiliz@gmail.com The neurofibromatoses are genetic disorders of the nervous system that primarily affect the development and growth of neural (nerve) cell tissues. The neurofibromatoses are classified as neurofibromatosis type 1 (NF1) and neurofibromatosis type 2 (NF2). NF1 is the more common type of the neurofibromatoses. The gene responsible for NF1 is located on the chromosome region 17q11.2 and for familial moyamoya disease on chromosome 17q25. This article reports on a 20-year-old female with neurofibromatosis-1 who developed moyamoya syndrome. More extensive reports and further investigations of such families having this combination will certainly provide a better understanding of this link in the near future. DOI: 10.1080/00207450801898279 PMID: 18576213 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16263726
1. J Biol Chem. 2005 Dec 23;280(51):41789-92. doi: 10.1074/jbc.C500395200. Epub 2005 Oct 31. Human but not yeast CHD1 binds directly and selectively to histone H3 methylated at lysine 4 via its tandem chromodomains. Sims RJ 3rd(1), Chen CF, Santos-Rosa H, Kouzarides T, Patel SS, Reinberg D. Author information: (1)Howard Hughes Medical Institute and Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA. Defining the protein factors that directly recognize post-translational, covalent histone modifications is essential toward understanding the impact of these chromatin "marks" on gene regulation. In the current study, we identify human CHD1, an ATP-dependent chromatin remodeling protein, as a factor that directly and selectively recognizes histone H3 methylated on lysine 4. In vitro binding studies identified that CHD1 recognizes di- and trimethyl H3K4 with a dissociation constant (Kd) of approximately 5 microm, whereas monomethyl H3K4 binds CHD1 with a 3-fold lower affinity. Surprisingly, human CHD1 binds to methylated H3K4 in a manner that requires both of its tandem chromodomains. In vitro analyses demonstrate that unlike human CHD1, yeast Chd1 does not bind methylated H3K4. Our findings indicate that yeast and human CHD1 have diverged in their ability to discriminate covalently modified histones and link histone modification-recognition and non-covalent chromatin remodeling activities within a single human protein. DOI: 10.1074/jbc.C500395200 PMCID: PMC1421377 PMID: 16263726 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17098252
1. J Mol Biol. 2007 Jan 26;365(4):1047-62. doi: 10.1016/j.jmb.2006.10.039. Epub 2006 Oct 14. Structural polymorphism of chromodomains in Chd1. Okuda M(1), Horikoshi M, Nishimura Y. Author information: (1)Graduate School of Supramolecular Biology, Yokohama City University, Tsurumi-ku, Yokohama 230-0045, Japan. Chromodomain from heterochromatin protein 1 and polycomb protein is known to be a lysine-methylated histone H3 tail-binding module. Chromo-helicase/ATPase DNA-binding protein 1 (CHD1) is an ATP-dependent chromatin remodeling factor, containing two tandem chromodomains. In human CHD1, both chromodomains are essential for specific binding to a K4 methylated histone H3 (H3 MeK4) peptide and are found to bind cooperatively in the crystal structure. For the budding yeast homologue, Chd1, the second but not the first chromodomain was once reported to bind to an H3 MeK4 peptide. Here, we reveal that neither the second chromodomain nor a region containing tandem chromodomains from yeast Chd1 bind to any lysine-methylated or arginine-methylated histone peptides that we examined. In addition, we examined the structures of the chromodomains from Chd1 by NMR. Although the tertiary structure of the region containing tandem chromodomains could not be obtained, the secondary structure deduced from NMR is well conserved in the tertiary structures of the corresponding first and second chromodomains determined individually by NMR. Both chromodomains of Chd1 demonstrate a structure similar to that of the corresponding part of CHD1, consisting of a three-stranded beta-sheet followed by a C-terminal alpha-helix. However, an additional helix between the first and second beta-strands, which is found in both of the first chromodomains of Chd1 and CHD1, is positioned in an entirely different manner in Chd1 and CHD1. In human CHD1 this helix forms the peptide-binding site. The amino acid sequences of the chromodomains could be well aligned on the basis of these structures. The alignment showed that yeast Chd1 lacks several key functional residues, which are responsible for specific binding to a methylated lysine residue in other chromodomains. Chd1 is likely to have no binding affinity for any H3 MeK peptide, as found in other chromodomain proteins. DOI: 10.1016/j.jmb.2006.10.039 PMID: 17098252 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/8733409
1. Tuber Lung Dis. 1996 Feb;77(1):19-26. doi: 10.1016/s0962-8479(96)90070-2. Clarithromycin against Mycobacterium avium complex infections. Heifets LB(1). Author information: (1)Department of Microbiology, University of Colorado Health Sciences Center, USA. The turning point in antimicrobial therapy of Mycobacterium avium infections came with the development of two new macrolides, clarithromycin and azithromycin. Controlled clinical trials, the first ever conducted with any agent among patients with M. avium infection, indicated the high efficiency of clarithromycin, in either acquired immune deficiency syndrome (AIDS) patients having a disseminated infection or non-AIDS patients with localized pulmonary disease. Monotherapy with clarithromycin resulted in elimination of bacteremia in almost all patients with disseminated infection, which is inevitably followed by a relapse of bacteremia in patients who survived long enough to reach this event. The strains susceptible to clarithromycin isolated before therapy contained 10(-8) or 10(-9) resistant mutants, and the relapses of bacteremia were caused by multiplication of these pre-existing mutants. Clarithromycin-resistance was associated with a mutation in the 23S rRNA gene. Cross-resistance between clarithromycin and azithromycin was confirmed with laboratory mutants and clinical isolates. At least two methods for determining the susceptibility of the M. avium isolates to clarithromycin are available: one is minimum inhibitory concentration (MIC) determination on Mueller-Hinton agar (pH 7.4) supplemented with 10% Oleic acid-albumin-dextrose catalase, the other is MIC determination in 7H12 broth, also at pH 7.4. The breakpoints for 'susceptible' for these methods are < or = 8.0 micrograms/ml and < or = 2.0 micrograms/ml, respectively. The breakpoints for 'resistant' are > 128 micrograms/ml for the agar method and > 32.0 micrograms/ml for the broth method. The predictability value of MIC determination was confirmed by comparing the test results with the patients' clinical and bacteriological response to therapy. The remaining major problem in the therapy of the M. avium infections is a selection of companion drugs to be used in combination with clarithromycin (or azithromycin) to prevent the emergence of the macrolide-resistance. A number of clinical trials are now in progress to find a solution to this problem. DOI: 10.1016/s0962-8479(96)90070-2 PMID: 8733409 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19335127
1. Neurosurg Focus. 2009 Apr;26(4):E11. doi: 10.3171/2009.1.FOCUS08310. Moyamoya disease: a summary. Burke GM(1), Burke AM, Sherma AK, Hurley MC, Batjer HH, Bendok BR. Author information: (1)New York Medical College, Valhalla, New York, USA. Moyamoya, meaning a "hazy puff of smoke" in Japanese, is a chronic, occlusive cerebrovascular disease involving bilateral stenosis or occlusion of the terminal portion of the internal carotid arteries (ICAs) and/or the proximal portions of the anterior cerebral arteries and middle cerebral arteries (MCAs). The Ministry of Health and Welfare of Japan has defined 4 types of moyamoya disease (MMD): ischemic, hemorrhagic, epileptic, and "other." The ischemic type has been shown to predominate in childhood, while the hemorrhagic type is more often observed in the adult population. The highest prevalence of MMD is found in Japan, with a higher female to male ratio. Studies have shown a possible genetic association of MMD linked to chromosome 17 in Japanese cases as well as in cases found in other demographics. During autopsy, intracerebral hematoma is found and most commonly serves as the major cause of death in patients with MMD. Moyamoya vessels at the base of the brain are composed of medium-sized or small muscular arteries emanating from the circle of Willis, mainly the intracranial portions of ICAs, anterior choroidal arteries, and posterior cerebral arteries, forming complex channels that connect with distal positions of the MCAs. Off of these channels are small tortuous and dilated vessels that penetrate into the base of the brain at the site of the thalamoperforate and lenticulostriate arteries. On angiography, there is the characteristic stenosis or occlusion bilaterally at the terminal portion of the ICAs as well as the moyamoya vessels at the base of the brain. Six angiographic stages have been described, from Stage 1, which reveals a narrowing of the carotid forks, to Stage 6, in which the moyamoya vessels disappear and collateral circulation is produced solely from the external carotid arteries. Cases with milder symptoms are usually treated conservatively; however, more severe symptomatic cases are treated using revascularization procedures. Surgical treatments are divided into 3 types: direct, indirect, and combined/other methods. Direct bypass includes superficial temporal artery-MCA bypass or use of other graft types. Indirect procedures bring in circulation to the intracranial regions by introducing newly developed vasculature from newly approximated tissues. These procedures may not be enough to prevent further ischemia; therefore, a combination of direct and indirect procedures is more suitable. This article will give a review of the epidemiology, natural history, pathology, pathophysiology, and diagnostic criteria, including imaging, and briefly describe the surgical treatment of MMD. DOI: 10.3171/2009.1.FOCUS08310 PMID: 19335127 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16584300
1. Microb Drug Resist. 2006 Spring;12(1):1-6. doi: 10.1089/mdr.2006.12.1. In vivo efficacy of phage therapy for Mycobacterium avium infection as delivered by a nonvirulent mycobacterium. Danelishvili L(1), Young LS, Bermudez LE. Author information: (1)Department of Biomedical Sciences, College of Veterinary Medicine, Department of Microbiology, College of Science, Oregon State University, 105 Magruder Hall, Corvallis, OR 97331-8797, USA. The emergence of mycobacteria resistant to currently available antimicrobial agents has become an important problem in modern medicine. Mycobacterium avium and M. tuberculosis are intracellular pathogens that replicate and survive within the mononuclear phagocytes. TM4 is a lytic mycobacteriophage that kills both extracellular M. avium and M. tuberculosis. When delivered by M. smegmatis transiently infected with TM4, it kills both M. avium and M. tuberculosis within RAW 264.7 macrophages. To evaluate the treatment of M. avium infection with phage in vivo, C57 BL/6 mice were infected with M. avium 109 and, 7 days later, treated either once or twice with TM4 phage (7.9 x 10(10) PFU/ml), M. smegmatis (4 x 10(8) cFU/ml), or M. smegmatis with TM4 phage delivered intravenously (i.v.). Treatment with TM4 phage alone or M. smegmatis without TM4 did not show a significant decrease in number of intracellular bacteria in the spleen compared with untreated control. In contrast, administration of M. smegmatis-TM4 resulted in a significant decrease in the number of M. avium in the spleen. However, 23% of bacteria recovered from treated mice were resistant to TM4. These in vivo studies confirmed the in vitro findings that an avirulent mycobacterium can be used as a carrier to deliver antimycobacterial phage intracellularly. DOI: 10.1089/mdr.2006.12.1 PMID: 16584300 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/1387133
1. J Antimicrob Chemother. 1992 Jun;29(6):693-9. doi: 10.1093/jac/29.6.693. TLC G-65 in combination with other agents in the therapy of Mycobacterium avium infection in beige mice. Cynamon MH(1), Klemens SP, Swenson CE. Author information: (1)Veterans Affairs Medical Center, Syracuse, New York. The activity of TLC G-65 (a liposomal gentamicin preparation), alone and in combination with rifapentine, clarithromycin, clofazimine and ethambutol, was evaluated in the beige mouse (C57BL/6J--bgj/bgj) model of disseminated Mycobacterium avium infection. TLC G-65 was found to be more active than amikacin. The combination of rifapentine and TLC G-65 was more active than either agent alone. The activity of clarithromycin in combination with TLC G-65 was similar to that of either agent alone. Clofazimine improved the activity of TLC G-65 with respect to the spleen, while ethambutol improved the activity with respect to the liver. Clofazimine and ethambutol enhanced the activity of TLC G-65 against bacteria in the lungs. TLC G-65 in combination with rifapentine appears to be an attractive regimen for the treatment of infections caused by bacteria in the M. avium complex. DOI: 10.1093/jac/29.6.693 PMID: 1387133 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19948887
1. Genetics. 2010 Feb;184(2):321-34. doi: 10.1534/genetics.109.111526. Epub 2009 Nov 30. Histone H3K4 and K36 methylation, Chd1 and Rpd3S oppose the functions of Saccharomyces cerevisiae Spt4-Spt5 in transcription. Quan TK(1), Hartzog GA. Author information: (1)Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz, California 95064, USA. Spt4-Spt5, a general transcription elongation factor for RNA polymerase II, also has roles in chromatin regulation. However, the relationships between these functions are not clear. Previously, we isolated suppressors of a Saccharomyces cerevisiae spt5 mutation in genes encoding members of the Paf1 complex, which regulates several cotranscriptional histone modifications, and Chd1, a chromatin remodeling enzyme. Here, we show that this suppression of spt5 can result from loss of histone H3 lysines 4 or 36 methylation, or reduced recruitment of Chd1 or the Rpd3S complex. These spt5 suppressors also rescue the synthetic growth defects observed in spt5 mutants that also lack elongation factor TFIIS. Using a FLO8 reporter gene, we found that a chd1 mutation caused cryptic initiation of transcription. We further observed enhancement of cryptic initiation in chd1 isw1 mutants and increased histone acetylation in a chd1 mutant. We suggest that, as previously proposed for H3 lysine 36 methylation and the Rpd3S complex, H3 lysine 4 methylation and Chd1 function to maintain normal chromatin structures over transcribed genes, and that one function of Spt4-Spt5 is to help RNA polymerase II overcome the repressive effects of these histone modifications and chromatin regulators on transcription. DOI: 10.1534/genetics.109.111526 PMCID: PMC2828714 PMID: 19948887 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18924029
1. Kardiol Pol. 2008 Sep;66(9):982-6; discussion 986. [Myopericarditis complicated with cardiogenic shock mimicking acute coronary syndrome with ST elevation in a patient with hyperthyroidism and diabetes mellitus]. [Article in Polish] Kukla P(1), Bryniarski L, Bromblik A, Szczuka K, Kawecka-Jaszcz K. Author information: (1)Oddział Wewnetrzny, Szpital Specjalistyczny im. H. Klimontowicza, Gorlice. kukla_piotr@poczta.onet.pl We describe a case of a 56 year old man with myopericarditis complicated with cardiogenic shock within first 3 days, mimicking on admission acute myocardial infarction with ST elevation in inferior ECG leads. Additionally, patient presented hyperthyroidism and totally decompensated diabetes mellitus. He required during the first 3 days intravenous infusion of inotropic agents. Cardiac enzymes levels were elevated. Akinesia in mid-inferior and mid-posterior regions in ECHO was observed. On the 10th day ST segment elevation in I, II, V3-V6 and ST depression in aVR was observed in ECG. After stabilisation patient underwent coronarography which showed normal coronary arteries. The final diagnosis was acute myopericarditis complicated with acute heart failure and cardiogenic shock. PMID: 18924029 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22422841
1. Nucleic Acids Res. 2012 Jul;40(12):e90. doi: 10.1093/nar/gks237. Epub 2012 Mar 15. Unveiling combinatorial regulation through the combination of ChIP information and in silico cis-regulatory module detection. Sun H(1), Guns T, Fierro AC, Thorrez L, Nijssen S, Marchal K. Author information: (1)Department of Microbial and Molecular Systems, Katholieke Universiteit Leuven, Leuven, Belgium. Computationally retrieving biologically relevant cis-regulatory modules (CRMs) is not straightforward. Because of the large number of candidates and the imperfection of the screening methods, many spurious CRMs are detected that are as high scoring as the biologically true ones. Using ChIP-information allows not only to reduce the regions in which the binding sites of the assayed transcription factor (TF) should be located, but also allows restricting the valid CRMs to those that contain the assayed TF (here referred to as applying CRM detection in a query-based mode). In this study, we show that exploiting ChIP-information in a query-based way makes in silico CRM detection a much more feasible endeavor. To be able to handle the large datasets, the query-based setting and other specificities proper to CRM detection on ChIP-Seq based data, we developed a novel powerful CRM detection method 'CPModule'. By applying it on a well-studied ChIP-Seq data set involved in self-renewal of mouse embryonic stem cells, we demonstrate how our tool can recover combinatorial regulation of five known TFs that are key in the self-renewal of mouse embryonic stem cells. Additionally, we make a number of new predictions on combinatorial regulation of these five key TFs with other TFs documented in TRANSFAC. DOI: 10.1093/nar/gks237 PMCID: PMC3384348 PMID: 22422841 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/7995828
1. J Cardiovasc Surg (Torino). 1994 Oct;35(5):383-9. Triiodothyronine administration in coronary artery bypass surgery: effect on hemodynamics. Vavouranakis I(1), Sanoudos G, Manios A, Kalogeropoulou K, Sitaras K, Kokkinos C. Author information: (1)Division of Cardiovascular Surgery, NIMTS Hospital, Athens, Greece. OBJECTIVE: The purpose of this study was to investigate any potential hemodynamic effect of intravenously administered triiodothyronine in patients undergoing coronary artery bypass surgery. EXPERIMENTAL DESIGN: Thirty patients were randomized in this single-blind, placebo-controlled trial. Hemodynamic parameters including heart rate, stroke volume index, cardiac index, pulmonary wedge pressure, pulmonary vascular resistances, systemic vascular resistances, and mean blood pressure, were compared between the two groups preoperatively, before the initiation of cardiopulmonary bypass (CPB), 5 minutes after the end of CPB, and 2, 4, 10, 16, and 22 hours thereafter. INTERVENTION: Triiodothyronine was administered as a bolus infusion over a 1 min period after removal of the aortic cross-clamp, (0.15 microgram/kg), before the end of CPB (0.1 microgram/kg), 4 hours after the end of CPB (0.1 microgram/kg), 9 hours after CPB (0.1 microgram/kg), and 14 hours after CPB (0.1 microgram/kg). Patients received inotropes, vasodilators, and diuretics only if specifically indicated. RESULTS: Plasma FT3 levels were higher in the T3 group, but within the normal range. No significant differences were noted in the pre and post CPB hemodynamics between the two groups for the most part of the study except that heart rate was increased in T3 group. A greater number of patients in the control group received vasodilators. No adverse reactions were noted with triiodothyronine administration. CONCLUSION: Triiodothyronine administration in patients undergoing cardiopulmonary bypass surgery is safe, may lessen the need for pharmacological (vasodilator) therapy, but may increase heart rate. PMID: 7995828 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12213743
1. Asian Cardiovasc Thorac Ann. 2002 Sep;10(3):219-22. doi: 10.1177/021849230201000306. Effects of intravenous triiodothyronine during coronary artery bypass surgery. Güden M(1), Akpinar B, Sagğbaş E, Sanisoğlu I, Cakali E, Bayindir O. Author information: (1)Department of Cardiovascular Surgery Kadir Has University, Florence Nightingale Hospital Istanbul, Turkey. A prospective randomized and double-blind study was performed to evaluate whether perioperative triiodothyronine administration has any effect on cardiovascular performance after coronary artery bypass surgery. Sixty patients were assigned to 2 groups of 30 each. When crossclamping ended, group A received an intravenous bolus of triiodothyronine, followed by infusion for 6 hours. Group B received a placebo. Serum triiodothyronine levels and hemo-dynamic parameters were serially measured. Mean postoperative cardiac index was slightly, but not significantly, higher in group A, whereas systemic vascular resistance was significantly lower in group A. Compared with preoperative values, serum triiodothyronine levels dropped significantly in group B at the end of cardiopulmonary bypass and remained low 12 hours postoperatively, while levels rose significantly in group A. No significant differences were detected between the groups in the incidence of arrhythmia, the need for inotropic support, intensive care unit stay, mortality, and morbidity. Perioperative administration of triiodothyronine increased cardiac output slightly and decreased systemic vascular resistance, but it had no effect on operative outcome. Routine use after coronary surgery is thus not recommended. DOI: 10.1177/021849230201000306 PMID: 12213743 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20671200
1. Proc Natl Acad Sci U S A. 2010 Aug 17;107(33):14615-20. doi: 10.1073/pnas.1002876107. Epub 2010 Jul 29. Genome-wide identification of cis-regulatory motifs and modules underlying gene coregulation using statistics and phylogeny. Rouault H(1), Mazouni K, Couturier L, Hakim V, Schweisguth F. Author information: (1)Laboratoire de Physique Statistique, Centre National de la Recherche Scientifique, Université Pierre et Marie Curie, Ecole Normale Supérieure, 75231, Paris Cedex 05, France. Cell fate determination depends in part on the establishment of specific transcriptional programs of gene expression. These programs result from the interpretation of the genomic cis-regulatory information by sequence-specific factors. Decoding this information in sequenced genomes is an important issue. Here, we developed statistical analysis tools to computationally identify the cis-regulatory elements that control gene expression in a set of coregulated genes. Starting with a small number of validated and/or predicted cis-regulatory modules (CRMs) in a reference species as a training set, but with no a priori knowledge of the factors acting in trans, we computationally predicted transcription factor binding sites (TFBSs) and genomic CRMs underlying coregulation. This method was applied to the gene expression program active in Drosophila melanogaster sensory organ precursor cells (SOPs), a specific type of neural progenitor cells. Mutational analysis showed that four, including one newly characterized, out of the five top-ranked families of predicted TFBSs were required for SOP-specific gene expression. Additionaly, 19 out of the 29 top-ranked predicted CRMs directed gene expression in neural progenitor cells, i.e., SOPs or larval brain neuroblasts, with a notable fraction active in SOPs (11/29). We further identified the lola gene as the target of two SOP-specific CRMs and found that the lola gene contributed to SOP specification. The statistics and phylogeny-based tools described here can be more generally applied to identify the cis-regulatory elements of specific gene regulatory networks in any family of related species with sequenced genomes. DOI: 10.1073/pnas.1002876107 PMCID: PMC2930411 PMID: 20671200 [Indexed for MEDLINE] Conflict of interest statement: The authors declare no conflict of interest.
http://www.ncbi.nlm.nih.gov/pubmed/23958074
1. J Cardiothorac Vasc Anesth. 2013 Dec;27(6):1218-23. doi: 10.1053/j.jvca.2013.01.027. Epub 2013 Aug 16. Efficacy of perioperative oral triiodothyronine replacement therapy in patients undergoing off-pump coronary artery bypass grafting. Choi YS(1), Shim JK, Song JW, Song Y, Yang SY, Kwak YL. Author information: (1)Department of Anesthesiology and Pain Medicine, Anesthesia and Pain Research Institute, Yonsei University College of Medicine. OBJECTIVE: The aim of this study was to assess the effects of oral triiodothyronine (T3) therapy on postoperative thyroid hormone concentrations, hemodynamic variables, and outcomes. DESIGN: A prospective, randomized, controlled, double-blind study. SETTING: Cardiac operating room at a single institution. PARTICIPANTS: One hundred patients undergoing elective off-pump coronary artery bypass graft surgery. INTERVENTIONS: Patients received either 20 μg of oral T3 or placebo every 12 hours starting 20 minutes before anesthetic induction, for a total of 4 doses. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of thyroid hormones were measured serially before surgery, upon arrival in the intensive care unit, and 12, 24, and 36 hours after surgery. Hemodynamic variables also were recorded serially. Postoperative inotrope requirement and major morbidity endpoints were assessed. Serum T3 concentrations were significantly higher with fewer patients having T3 concentrations below the normal range in the T3 group than the placebo group throughout the postoperative period. Hemodynamic variables, postoperative inotrope requirement, and outcome variables showed no differences between the groups. CONCLUSIONS: Oral T3 therapy significantly attenuated the postoperative decline in T3 concentrations in patients undergoing off-pump coronary artery bypass graft surgery. The lack of apparent clinical benefit merits further investigations in patients with reduced cardiac performance. Copyright © 2013 Elsevier Inc. All rights reserved. DOI: 10.1053/j.jvca.2013.01.027 PMID: 23958074 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/10343261
1. J Thorac Cardiovasc Surg. 1999 Jun;117(6):1128-34. doi: 10.1016/s0022-5223(99)70249-7. A randomized double-blind study of the effect of triiodothyronine on cardiac function and morbidity after coronary bypass surgery. Mullis-Jansson SL(1), Argenziano M, Corwin S, Homma S, Weinberg AD, Williams M, Rose EA, Smith CR. Author information: (1)Departments of Anesthesiology, Surgery,and Medicine, Columbia University College of Physicians and Surgeons, New York, NY, USA. BACKGROUND: Although triiodothyronine deficiency has been described after cardiopulmonary bypass, data supporting its use have been conflicting. A double-blind, randomized, placebo-controlled study was undertaken to further define the effect of triiodothyronine on hemodynamics and outcome after coronary artery bypass grafting. METHODS: A total of 170 patients undergoing elective coronary artery bypass grafting were enrolled and completed the study from November 1996 through March 1998. On removal of the aortic crossclamp, patients were randomized to receive either intravenous triiodothyronine (0.4 microgram/kg bolus plus 0.1 microgram/kg infusion administered over a 6-hour period, n = 81) or placebo (n = 89). Outcome variables included hemodynamic profile and inotropic drug/pressor requirements at several time points (mean +/- standard error of the mean), perioperative morbidity (arrhythmia/ischemia/infarction), and mortality. RESULTS: Despite similar baseline characteristics, patients randomized to triiodothyronine had a higher cardiac index and lower inotropic requirements after the operation. Subjects receiving triiodothyronine demonstrated a significantly lower incidence of postoperative myocardial ischemia (4% vs 18%, P =.007) and pacemaker dependence (14% vs 25%, P =.013). Seven patients in the placebo group required postoperative mechanical assistance (intra-aortic balloon pump, n = 4; left ventricular assist device, n = 3), compared with none in the triiodothyronine group (P =.01). There were 2 deaths in the placebo group and no deaths in the triiodothyronine group. CONCLUSIONS: Parenteral triiodothyronine given after crossclamp removal during elective coronary artery bypass grafting significantly improved postoperative ventricular function, reduced the need for treatment with inotropic agents and mechanical devices, and decreased the incidence of myocardial ischemia. The incidence of atrial fibrillation was slightly decreased, and the need for postoperative pacemaker support was reduced. DOI: 10.1016/s0022-5223(99)70249-7 PMID: 10343261 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18606616
1. Nucleic Acids Res. 2008 Aug;36(13):4488-97. doi: 10.1093/nar/gkn407. Epub 2008 Jul 7. MOPAT: a graph-based method to predict recurrent cis-regulatory modules from known motifs. Hu J(1), Hu H, Li X. Author information: (1)Division of Biostatistics, School of Informatics, Indiana University, 410 West 10th Street, Indianapolis, IN 46202, USA. The identification of cis-regulatory modules (CRMs) can greatly advance our understanding of eukaryotic regulatory mechanism. Current methods to predict CRMs from known motifs either depend on multiple alignments or can only deal with a small number of known motifs provided by users. These methods are problematic when binding sites are not well aligned in multiple alignments or when the number of input known motifs is large. We thus developed a new CRM identification method MOPAT (motif pair tree), which identifies CRMs through the identification of motif modules, groups of motifs co-occurring in multiple CRMs. It can identify 'orthologous' CRMs without multiple alignments. It can also find CRMs given a large number of known motifs. We have applied this method to mouse developmental genes, and have evaluated the predicted CRMs and motif modules by microarray expression data and known interacting motif pairs. We show that the expression profiles of the genes containing CRMs of the same motif module correlate significantly better than those of a random set of genes do. We also show that the known interacting motif pairs are significantly included in our predictions. Compared with several current methods, our method shows better performance in identifying meaningful CRMs. DOI: 10.1093/nar/gkn407 PMCID: PMC2490743 PMID: 18606616 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/6966005
1. Johns Hopkins Med J. 1980 Apr;146(4):133-6. Haemophilus influenzae sepsis leading to pericarditis despite antimicrobial therapy. Leggiadro RJ, Balsam D. Acute purulent pericarditis is a well-recognized, though infrequently seen, manifestation of systemic Haemophilus influenzae type b disease. We recently studied two pediatric patients who developed signs of this septic complication during appropriate antibiotic treatment for bacteremia. These case reports should alert physicians to the possibility that pericarditis may become clinically evident in patients with systemic H. influenzae infections many days after initiation of appropriate therapy. The pathophysiology, diagnostic modalities and therapy are briefly reviewed. PMID: 6966005 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16315865
1. Gan To Kagaku Ryoho. 2005 Oct;32(11):1550-2. [Evaluation of host immunity and side effects in breast cancer patients treated with adjuvant chemotherapy (FEC therapy)]. [Article in Japanese] Nagashima Y(1), Sanpei N, Yamamoto S, Yoshino S, Tangoku A, Oka M. Author information: (1)Dept. of Surgery II, Yamaguchi University School of Medicine. BACKGROUND: FEC (5-FU+epirubicin+cyclophosphamide) therapy has been used as adjuvant chemotherapy for breast cancer patients with nodes positive. The aim of this study was to evaluate host immunity and side effects of the FEC therapy. The effect of oral administration of Lentinus edodes mycelia (LEM) was also observed. METHODS: Ten patients were enrolled in this study. The treatment with 5-FU (500 mg/m2), epirubicin (75 mg/m2) and cyclophosphamide (500 mg/m2) was administered every 21 days for 2 cycles, and LEM (9 g/day po) was administered during the 2nd cycle. RESULTS: NK cell activity and the number of white blood cells decreased on the 7th day after the therapy, and they recovered on the 21st day. However, this NK cell activity and the number of white blood cells didn't decrease when the FEC therapy was used with LEM po. CONCLUSIONS: FEC 75 therapy has made some impacts on host immunity, and LEM with the FEC 75 therapy might have prevented host immunity. PMID: 16315865 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12525522
1. J Clin Oncol. 2003 Jan 15;21(2):298-305. doi: 10.1200/JCO.2003.04.148. Randomized trial comparing six versus three cycles of epirubicin-based adjuvant chemotherapy in premenopausal, node-positive breast cancer patients: 10-year follow-up results of the French Adjuvant Study Group 01 trial. Fumoleau P(1), Kerbrat P, Romestaing P, Fargeot P, Brémond A, Namer M, Schraub S, Goudier MJ, Mihura J, Monnier A, Clavère P, Serin D, Seffert P, Pourny C, Facchini T, Jacquin JP, Sztermer JF, Datchary J, Ramos R, Luporsi E. Author information: (1)Département d'Oncologie Médicale, Centre René Gauducheau, Nantes, France. fumoleau@gauducheau-nantes.fnclcc.fr PURPOSE: To evaluate the duration and dose intensity of epirubicin-based regimens in premenopausal patients with lymph node-positive breast cancer. PATIENTS AND METHODS: Between 1986 and 1990, 621 patients with operable breast cancer were randomly assigned to receive fluorouracil (Roche SA, Basel, Switzerland) 500 mg/m2, epirubicin (Pharmacia SA, Milan, Italy) 50 mg/m2, and cyclophosphamide (Asta Medica AG, Frankfurt, Germany) 500 mg/m2 every 21 days (FEC 50) for six cycles (6 FEC 50); FEC 50 for three cycles (3 FEC 50); or the same regimen with epirubicin 75 mg/m2 (FEC 75) for three cycles (3 FEC 75). All patients in the three arms received chest wall irradiation at the end of the third cycle. RESULTS: After a 131-month median follow-up, the 10-year disease-free survival (DFS) was 53.4%, 42.5%, and 43.6% (P =.05) in the three arms, respectively. Pairwise comparisons demonstrate that 6 FEC 50 was superior both to 3 FEC 50 (P =.02) and to 3 FEC 75 (P =.05). The 10-year overall survival (OS) for the 6 FEC 50 arm was 64.3%, for the 3 FEC 50 arm it was 56.6%, and for the 3 FEC 75 arm, it was 59.7% (P =.25), respectively. Pairwise comparisons demonstrate that 6 FEC 50 was more effective than 3 FEC 50 (P =.10). Cox regression analysis demonstrates that OS was significantly better in the 6 FEC 50 than in the 3 FEC 50 arm (P =.046). No severe infections (grade 3 to 4), acute cardiac toxicity, or deaths from toxicity have been observed. Only five patients developed delayed cardiac dysfunctions, and three patients developed acute myeloblastic leukemia. CONCLUSION: After a long-term follow-up in an adjuvant setting, the benefit of six cycles of FEC 50 compared with three cycles, whatever the dose, is highly significant in terms of DFS. As regards OS, the group receiving six cycles of FEC 50 has significantly better results than the group receiving three cycles of FEC 50. DOI: 10.1200/JCO.2003.04.148 PMID: 12525522 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/8808724
1. J Chemother. 1996 Jun;8(3):237-42. doi: 10.1179/joc.1996.8.3.237. Prevention of delayed emesis by a single intravenous bolus dose of 5-HT3-receptor-antagonist in moderately emetogenic chemotherapy. Massidda B(1), Ionta MT. Author information: (1)Division of Medical Oncology, University of Cagliari, Italy. While the use of 5-HT3 receptor antagonists is clearly justified in patients receiving cisplatin, their role with less emetic drugs is still not defined. The aim of our randomized study was to verify the efficacy of the single standard dose of three 5-HT3-receptor-antagonists in moderately emetic chemotherapies. Sixty chemotherapy-naive breast cancer patients of 30 to 71 years in age, P.S. = 0-1, receiving 5-fluorouracil-epirubicin-cyclophosphamide (FEC 75) q 21 days or cyclophosphamide-methotrexate-5-fluorouracil (CMF) or 120 mg/m2 epirubicin or high dose mitomycin-methotrexate-mitoxantrone (MMM) q 14 days (+ G-CSF) or 100 mg/m2 epirubicin (+ G-CSF) were randomized to receive, 15 min before chemotherapy, 8 mg i.v. bolus of ondansetron or 3 mg i.v. granisetron or 5 mg i.v. tropisetron and no further antiemetic therapy in the following days. 180 cycles were evaluable. Complete protection, (the absence of vomiting episodes,) was respectively 75%, 70% and 70% in the acute and 70%, 82%, 72% in the delayed phases, and an absence of nausea was 56%, 37% and 20% in the acute phase and 50%, 35% and 27% in the delayed, respectively. Complete response, (absence of vomiting and absence or mild nausea,) was 74%, 58.6% and 50.8% in the acute and 64%, 63.7%, 47.3% in the delayed phases, respectively. At the statistical analysis no significant differences between the three drugs were found regarding acute vomiting while ondansetron was superior to granisetron and tropisetron in acute (p = 0.018; p < 0.05) and delayed nausea (P = 0.104; p < 0.01). This activity is practically the same as that we reported (Ann Oncol 1994; 6, suppl 8: 204) with a loading dose on day 1 and maintenance for the following 2-5 days, but with a significantly favorable cost-benefit ratio. DOI: 10.1179/joc.1996.8.3.237 PMID: 8808724 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22874562
1. Autophagy. 2012 Nov;8(11):1677-9. doi: 10.4161/auto.21484. Epub 2012 Aug 9. Inactivation of the Cullin (CUL)-RING E3 ligase by the NEDD8-activating enzyme inhibitor MLN4924 triggers protective autophagy in cancer cells. Luo Z(1), Pan Y, Jeong LS, Liu J, Jia L. Author information: (1)Department of Immunology, Shanghai Medical College, Fudan University, Shanghai, China. Comment on Luo Z, Yu G, Lee HW, Li L, Wang L, Yang D, et al. The Nedd8-activating enzyme inhibitor MLN4924 induces autophagy and apoptosis to suppress liver cancer cell growth. Cancer Res. 2012;72:3360–71. doi: 10.1158/0008-5472.CAN-12-0388. The multiunit Cullin (CUL)-RING E3 ligase (CRL) controls diverse biological processes by targeting a mass of substrates for ubiquitination and degradation, whereas its dysfunction causes carcinogenesis. Post-translational neddylation of CUL, a process triggered by the NEDD8-activating enzyme E1 subunit 1 (NAE1), is required for CRL activation. Recently, MLN4924 was discovered via a high-throughput screen as a specific NAE1 inhibitor and first-in-class anticancer drug. By blocking CUL neddylation, MLN4924 inactivates CRL and causes the accumulation of CRL substrates that trigger cell cycle arrest, senescence and/or apoptosis to suppress the growth of cancer cells in vitro and in vivo. Recently, we found that MLN4924 also triggers protective autophagy in response to CRL inactivation. MLN4924-induced autophagy is attributed partially to the inhibition of mechanistic target of rapamycin (also known as mammalian target of rapamycin, MTOR) activity by the accumulation of the MTOR inhibitory protein DEPTOR, as well as reactive oxygen species (ROS)-induced stress. Moreover, the blockage of autophagy response enhances apoptosis in MLN4924-treated cells. Together, our findings not only reveal autophagy as a novel cellular response to CRL inactivation by MLN4924, but also provide a piece of proof-of-concept evidence for the combination of MLN4924 with autophagy inhibitors to enhance therapeutic efficacy. DOI: 10.4161/auto.21484 PMCID: PMC3494597 PMID: 22874562 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11875727
1. Br J Cancer. 2002 Mar 4;86(5):692-7. doi: 10.1038/sj.bjc.6600165. Sequential or alternating administration of docetaxel (Taxotere) combined with FEC in metastatic breast cancer: a randomised phase II trial. Spielmann M(1), Tubiana-Hulin M, Namer M, Mansouri H, Bougnoux Ph, Tubiana-Mathieu N, Lotz V, Eymard JC. Author information: (1)Institut Gustave Roussy, 39-53 rue Camille Desmoulins, 94805 Villejuif, France. spielman@igr.fr The aim of this study, using a Fleming single-stage design, was to explore the efficacy and safety of Taxotere 100 x mg x m(-2) docetaxel and FEC 75 cyclophosphamide 500 mg x m(-2), fluorouracil 500 x mg x m(-2) and epirubicin 75 mg x m(-2), in alternating and sequential schedules for the first-line treatment of metastatic breast cancer. One hundred and thirty-six women were randomly allocated, to one of three treatment regimens: DTX 100 plus FEC 75, alternated for eight courses (ALT); four courses of DTX 100 followed by four courses of FEC 75 (SEQ T); or four courses of FEC 75 followed by four courses of DTX 100 (SEQ F). One hundred and thirty-one women were evaluable for tumour response. Although the treatment outcome was equivalent in the two sequential arms and the alternating regimen (P=0.110, not significant), the response rate was less encouraging in the SEQ F arm (52.3%) than in the other two arms (71.1% for ALT and 70.5% for SEQ T), in which docetaxel was administered first. Time to progression was similar in the ALT, SEQ T and SEQ F arms (9.5, 9.3 and 10.4 months respectively). Grade 3-4 neutropenia was observed in nearly all patients; febrile neutropenia occurred in 9% (ALT), 16% (SEQ T) and 2% (SEQ F) of patients. Few patients (< or =9%) developed grade 3-4 non-haematological toxicities. Relative dose intensity was 97-99% for all regimens. All treatment regimens were active and well tolerated. Copyright 2002 Cancer Research UK DOI: 10.1038/sj.bjc.6600165 PMCID: PMC2375306 PMID: 11875727 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22072567
1. Cancer Res. 2012 Jan 1;72(1):282-93. doi: 10.1158/0008-5472.CAN-11-2866. Epub 2011 Nov 9. Radiosensitization of human pancreatic cancer cells by MLN4924, an investigational NEDD8-activating enzyme inhibitor. Wei D(1), Li H, Yu J, Sebolt JT, Zhao L, Lawrence TS, Smith PG, Morgan MA, Sun Y. Author information: (1)Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, Michigan 48109, USA. Radiotherapy is used in locally advanced pancreatic cancers in which it can improve survival in combination with gemcitabine. However, prognosis is still poor in this setting in which more effective therapies remain needed. MLN4924 is an investigational small molecule currently in phase I clinical trials. MLN4924 inhibits NAE (NEDD8 Activating Enzyme), a pivotal regulator of the E3 ubiquitin ligase SCF (SKP1, Cullins, and F-box protein), that has been implicated recently in DNA damage and repair. In this study, we provide evidence that MLN4924 can be used as an effective radiosensitizer in pancreatic cancer. Specifically, MLN4924 (20-100 nmol/L) effectively inhibited cullin neddylation and sensitized pancreatic cancer cells to ionizing radiation in vitro with a sensitivity enhancement ratio of approximately 1.5. Mechanistically, MLN4924 treatment stimulated an accumulation of several SCF substrates, including CDT1, WEE1, and NOXA, in parallel with an enhancement of radiation-induced DNA damage, aneuploidy, G(2)/M phase cell-cycle arrest, and apoptosis. RNAi-mediated knockdown of CDT1 and WEE1 partially abrogated MLN4924-induced aneuploidy, G(2)/M arrest, and radiosensitization, indicating a causal effect. Furthermore, MLN4924 was an effective radiosensitizer in a mouse xenograft model of human pancreatic cancer. Our findings offer proof-of-concept for use of MLN4924 as a novel class of radiosensitizer for the treatment of pancreatic cancer. ©2011 AACR. DOI: 10.1158/0008-5472.CAN-11-2866 PMCID: PMC3251739 PMID: 22072567 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19360080
1. Nature. 2009 Apr 9;458(7239):732-6. doi: 10.1038/nature07884. An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer. Soucy TA(1), Smith PG, Milhollen MA, Berger AJ, Gavin JM, Adhikari S, Brownell JE, Burke KE, Cardin DP, Critchley S, Cullis CA, Doucette A, Garnsey JJ, Gaulin JL, Gershman RE, Lublinsky AR, McDonald A, Mizutani H, Narayanan U, Olhava EJ, Peluso S, Rezaei M, Sintchak MD, Talreja T, Thomas MP, Traore T, Vyskocil S, Weatherhead GS, Yu J, Zhang J, Dick LR, Claiborne CF, Rolfe M, Bolen JB, Langston SP. Author information: (1)Discovery, Millennium Pharmaceuticals, Inc., 40 Landsdowne Street, Cambridge, Massachusetts 02139, USA. teresa.soucy@mpi.com Comment in Nature. 2009 Apr 9;458(7239):709-10. doi: 10.1038/458709a. The clinical development of an inhibitor of cellular proteasome function suggests that compounds targeting other components of the ubiquitin-proteasome system might prove useful for the treatment of human malignancies. NEDD8-activating enzyme (NAE) is an essential component of the NEDD8 conjugation pathway that controls the activity of the cullin-RING subtype of ubiquitin ligases, thereby regulating the turnover of a subset of proteins upstream of the proteasome. Substrates of cullin-RING ligases have important roles in cellular processes associated with cancer cell growth and survival pathways. Here we describe MLN4924, a potent and selective inhibitor of NAE. MLN4924 disrupts cullin-RING ligase-mediated protein turnover leading to apoptotic death in human tumour cells by a new mechanism of action, the deregulation of S-phase DNA synthesis. MLN4924 suppressed the growth of human tumour xenografts in mice at compound exposures that were well tolerated. Our data suggest that NAE inhibitors may hold promise for the treatment of cancer. DOI: 10.1038/nature07884 PMID: 19360080 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24672057
1. Mol Cancer Ther. 2014 Jun;13(6):1625-35. doi: 10.1158/1535-7163.MCT-13-0634. Epub 2014 Mar 26. Nedd8-activating enzyme inhibitor MLN4924 provides synergy with mitomycin C through interactions with ATR, BRCA1/BRCA2, and chromatin dynamics pathways. Garcia K(1), Blank JL(1), Bouck DC(1), Liu XJ(1), Sappal DS(1), Hather G(1), Cosmopoulos K(1), Thomas MP(1), Kuranda M(1), Pickard MD(1), Liu R(1), Bandi S(1), Smith PG(1), Lightcap ES(2). Author information: (1)Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts. (2)Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts eric.lightcap@takeda.com. MLN4924 is an investigational small-molecule inhibitor of the Nedd8-activating enzyme currently in phase I clinical trials. MLN4924 induces DNA damage via rereplication in most cell lines. This distinct mechanism of DNA damage may affect its ability to combine with standard-of-care agents and may affect the clinical development of MLN4924. As such, we studied its interaction with other DNA-damaging agents. Mitomycin C, cisplatin, cytarabine, UV radiation, SN-38, and gemcitabine demonstrated synergy in combination with MLN4924 in vitro. The combination of mitomycin C and MLN4924 was shown to be synergistic in a mouse xenograft model. Importantly, depletion of genes within the ataxia telangiectasia and Rad3 related (ATR) and BRCA1/BRCA2 pathways, chromatin modification, and transcription-coupled repair reduced the synergy between mitomycin C and MLN4924. In addition, comet assay demonstrated increased DNA strand breaks with the combination of MLN4924 and mitomycin C. Our data suggest that mitomycin C causes stalled replication forks, which when combined with rereplication induced by MLN4924 results in frequent replication fork collisions, leading to cell death. This study provides a straightforward approach to understand the mechanism of synergy, which may provide useful information for the clinical development of these combinations. ©2014 American Association for Cancer Research. DOI: 10.1158/1535-7163.MCT-13-0634 PMID: 24672057 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21417215
1. J Org Chem. 2011 May 6;76(9):3557-61. doi: 10.1021/jo2001897. Epub 2011 Mar 21. Stereoselective synthesis of MLN4924, an inhibitor of NEDD8-activating enzyme. Lee HW(1), Nam SK, Choi WJ, Kim HO, Jeong LS. Author information: (1)Department of Bioinspired Science and Laboratory of Medicinal Chemistry, College of Pharmacy, Ewha Womans University, Seoul 120-750, Korea. Erratum in J Org Chem. 2011 May 6;76(9):3614. MLN4924 (1), which is in clinical trials as an anticancer agent, was stereoselectively synthesized from d-ribose via a route involving stereoselective reduction, regioselective cleavage of an isopropylidene moiety, and selective displacement of a cyclic sulfate moiety as key steps. © 2011 American Chemical Society DOI: 10.1021/jo2001897 PMID: 21417215 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21463634
1. J Mol Biol. 2011 Jun 3;409(2):136-45. doi: 10.1016/j.jmb.2011.03.023. Epub 2011 Apr 2. Neddylation-induced conformational control regulates cullin RING ligase activity in vivo. Boh BK(1), Smith PG, Hagen T. Author information: (1)NUS Graduate School for Integrative Sciences and Engineering (NGS), National University of Singapore, Singapore 117456, Singapore. Cullin RING ligases (CRLs) constitute the largest family of ubiquitin ligases with diverse cellular functions. Conjugation of the ubiquitin-like molecule Nedd8 to a conserved lysine residue on the cullin scaffold is essential for the activity of CRLs. Using structural studies and in vitro assays, it has been demonstrated that neddylation stimulates CRL activity through conformational rearrangement of the cullin C-terminal winged-helix B domain and Rbx1 RING subdomain from a closed architecture to an open and dynamic structure, thus promoting ubiquitin transfer onto the substrate. Here, we tested whether the proposed mechanism operates in vivo in intact cells and applies to other CRL family members. To inhibit cellular neddylation, we used a cell line with tetracycline-inducible expression of a dominant-negative form of the Nedd8 E2 enzyme or treatment of cells with the Nedd8 E1 inhibitor MLN4924. Using these cellular systems, we show that different mutants of Cul2 and Cul3 and of Rbx1 that confer increased Rbx1 flexibility mimic neddylation and rescue CRL activity in intact cells. Our findings indicate that in vivo neddylation functions by inducing conformational changes in the C-terminal domain of Cul2 and Cul3 that free the RING domain of Rbx1 and bridge the gap for ubiquitin transfer onto the substrate. Copyright © 2011 Elsevier Ltd. All rights reserved. DOI: 10.1016/j.jmb.2011.03.023 PMID: 21463634 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22246439
1. Mol Cancer Ther. 2012 Apr;11(4):942-51. doi: 10.1158/1535-7163.MCT-11-0563. Epub 2012 Jan 12. Molecular and cellular effects of NEDD8-activating enzyme inhibition in myeloma. McMillin DW(1), Jacobs HM, Delmore JE, Buon L, Hunter ZR, Monrose V, Yu J, Smith PG, Richardson PG, Anderson KC, Treon SP, Kung AL, Mitsiades CS. Author information: (1)Department of Medical Oncology, Jerome Lipper Multiple Myeloma Center, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02215, USA. The NEDD8-activating enzyme is upstream of the 20S proteasome in the ubiquitin/proteasome pathway and catalyzes the first step in the neddylation pathway. NEDD8 modification of cullins is required for ubiquitination of cullin-ring ligases that regulate degradation of a distinct subset of proteins. The more targeted impact of NEDD8-activating enzyme on protein degradation prompted us to study MLN4924, an investigational NEDD8-activating enzyme inhibitor, in preclinical multiple myeloma models. In vitro treatment with MLN4924 led to dose-dependent decrease of viability (EC(50) = 25-150 nmol/L) in a panel of human multiple myeloma cell lines. MLN4924 was similarly active against a bortezomib-resistant ANBL-6 subline and its bortezomib-sensitive parental cells. MLN4924 had submicromolar activity (EC(50) values <500 nmol/L) against primary CD138(+) multiple myeloma patient cells and exhibited at least additive effect when combined with dexamethasone, doxorubicin, and bortezomib against MM.1S cells. The bortezomib-induced compensatory upregulation of transcripts for ubiquitin/proteasome was not observed with MLN4924 treatment, suggesting distinct functional roles of NEDD8-activating enzyme versus 20S proteasome. MLN4924 was well tolerated at doses up to 60 mg/kg 2× daily and significantly reduced tumor burden in both a subcutaneous and an orthotopic mouse model of multiple myeloma. These studies provide the framework for the clinical investigation of MLN4924 in multiple myeloma. DOI: 10.1158/1535-7163.MCT-11-0563 PMCID: PMC3755358 PMID: 22246439 [Indexed for MEDLINE] Conflict of interest statement: Disclosure of Potential Conflicts of Interest DWM is a founder and equity holder in Axios Biosciences. HMJ, JD, LB, ZH, VM, ALK have nothing to disclose. JY, PG are employees of Millennium Pharmaceuticals (The Takeda Oncology company); P.G.R. has received honoraria from Millennium, Celgene and Novartis; as well as research support from Johnson & Johnson and Bristol-Myers Squibb. K.C.A. has received research grants and honoraria from Millennium and Celgene, has been a consultant for Millennium, Celgene, Novartis, Bristol-Myers Squibb, Merck and Onyx and is a founder of Acetylon Pharmaceutials. SPT has received honoraria from and is consultant/member of advisory board for Millennium. CSM has received in the past Consultant honoraria from Millennium Pharmaceuticals, Novartis Pharmaceuticals, Bristol-Myers Squibb, Merck & Co., Kosan Pharmaceuticals, Pharmion, Johnson & Johnson and Amnis Therapeutics, licensing royalties from PharmaMar, and research funding from Amgen Pharmaceuticals, AVEO Pharma, EMD Serono, Sunesis, Gloucester Pharmaceuticals and Johnson & Johnson.
http://www.ncbi.nlm.nih.gov/pubmed/19920172
1. Proc Natl Acad Sci U S A. 2009 Dec 8;106(49):20776-81. doi: 10.1073/pnas.0906998106. Epub 2009 Nov 17. The anti-apoptotic protein HAX-1 is a regulator of cardiac function. Zhao W(1), Waggoner JR, Zhang ZG, Lam CK, Han P, Qian J, Schroder PM, Mitton B, Kontrogianni-Konstantopoulos A, Robia SL, Kranias EG. Author information: (1)Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0575, USA. The HS-1 associated protein X-1 (HAX-1) is a ubiquitously expressed protein that protects cardiomyocytes from programmed cell death. Here we identify HAX-1 as a regulator of contractility and calcium cycling in the heart. HAX-1 overexpression reduced sarcoplasmic reticulum Ca-ATPase (SERCA2) pump activity in isolated cardiomyocytes and in vivo, leading to depressed myocyte calcium kinetics and mechanics. Conversely, downregulation of HAX-1 enhanced calcium cycling and contractility. The inhibitory effects of HAX-1 were abolished upon phosphorylation of phospholamban, which plays a fundamental role in controlling basal contractility and constitutes a key downstream effector of the beta-adrenergic signaling cascade. Mechanistically, HAX-1 promoted formation of phospholamban monomers, the active/inhibitory units of the calcium pump. Indeed, ablation of PLN rescued HAX-1 inhibition of contractility in vivo. Thus, HAX-1 represents a regulatory mechanism in cardiac calcium cycling and its responses to sympathetic stimulation, implicating its importance in calcium homeostasis and cell survival. DOI: 10.1073/pnas.0906998106 PMCID: PMC2791603 PMID: 19920172 [Indexed for MEDLINE] Conflict of interest statement: The authors declare no conflict of interest.
http://www.ncbi.nlm.nih.gov/pubmed/18971376
1. Mol Biol Cell. 2009 Jan;20(1):306-18. doi: 10.1091/mbc.e08-06-0587. Epub 2008 Oct 29. The anti-apoptotic protein HAX-1 interacts with SERCA2 and regulates its protein levels to promote cell survival. Vafiadaki E(1), Arvanitis DA, Pagakis SN, Papalouka V, Sanoudou D, Kontrogianni-Konstantopoulos A, Kranias EG. Author information: (1)Molecular Biology Division, Biomedical Research Foundation, Academy of Athens, Greece. Cardiac contractility is regulated through the activity of various key Ca(2+)-handling proteins. The sarco(endo)plasmic reticulum (SR) Ca(2+) transport ATPase (SERCA2a) and its inhibitor phospholamban (PLN) control the uptake of Ca(2+) by SR membranes during relaxation. Recently, the antiapoptotic HS-1-associated protein X-1 (HAX-1) was identified as a binding partner of PLN, and this interaction was postulated to regulate cell apoptosis. In the current study, we determined that HAX-1 can also bind to SERCA2. Deletion mapping analysis demonstrated that amino acid residues 575-594 of SERCA2's nucleotide binding domain are required for its interaction with the C-terminal domain of HAX-1, containing amino acids 203-245. In transiently cotransfected human embryonic kidney 293 cells, recombinant SERCA2 was specifically targeted to the ER, whereas HAX-1 selectively concentrated at mitochondria. On triple transfections with PLN, however, HAX-1 massively translocated to the ER membranes, where it codistributed with PLN and SERCA2. Overexpression of SERCA2 abrogated the protective effects of HAX-1 on cell survival, after hypoxia/reoxygenation or thapsigargin treatment. Importantly, HAX-1 overexpression was associated with down-regulation of SERCA2 expression levels, resulting in significant reduction of apparent ER Ca(2+) levels. These findings suggest that HAX-1 may promote cell survival through modulation of SERCA2 protein levels and thus ER Ca(2+) stores. DOI: 10.1091/mbc.e08-06-0587 PMCID: PMC2613088 PMID: 18971376 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20129059
1. Mol Cell. 2010 Jan 15;37(1):102-11. doi: 10.1016/j.molcel.2009.12.024. Substrate-assisted inhibition of ubiquitin-like protein-activating enzymes: the NEDD8 E1 inhibitor MLN4924 forms a NEDD8-AMP mimetic in situ. Brownell JE(1), Sintchak MD, Gavin JM, Liao H, Bruzzese FJ, Bump NJ, Soucy TA, Milhollen MA, Yang X, Burkhardt AL, Ma J, Loke HK, Lingaraj T, Wu D, Hamman KB, Spelman JJ, Cullis CA, Langston SP, Vyskocil S, Sells TB, Mallender WD, Visiers I, Li P, Claiborne CF, Rolfe M, Bolen JB, Dick LR. Author information: (1)Discovery, Millennium Pharmaceuticals, Inc., 40 Landsdowne Street, Cambridge, MA 02139, USA. The NEDD8-activating enzyme (NAE) initiates a protein homeostatic pathway essential for cancer cell growth and survival. MLN4924 is a selective inhibitor of NAE currently in clinical trials for the treatment of cancer. Here, we show that MLN4924 is a mechanism-based inhibitor of NAE and creates a covalent NEDD8-MLN4924 adduct catalyzed by the enzyme. The NEDD8-MLN4924 adduct resembles NEDD8 adenylate, the first intermediate in the NAE reaction cycle, but cannot be further utilized in subsequent intraenzyme reactions. The stability of the NEDD8-MLN4924 adduct within the NAE active site blocks enzyme activity, thereby accounting for the potent inhibition of the NEDD8 pathway by MLN4924. Importantly, we have determined that compounds resembling MLN4924 demonstrate the ability to form analogous adducts with other ubiquitin-like proteins (UBLs) catalyzed by their cognate-activating enzymes. These findings reveal insights into the mechanism of E1s and suggest a general strategy for selective inhibition of UBL conjugation pathways. Copyright 2010 Elsevier Inc. All rights reserved. DOI: 10.1016/j.molcel.2009.12.024 PMID: 20129059 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24213570
1. Oncogene. 2014 Oct 30;33(44):5211-20. doi: 10.1038/onc.2013.473. Epub 2013 Nov 11. Endothelial deletion of Sag/Rbx2/Roc2 E3 ubiquitin ligase causes embryonic lethality and blocks tumor angiogenesis. Tan M(1), Li H(1), Sun Y(1). Author information: (1)Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, MI, USA. SAG (Sensitive to Apoptosis Gene), also known as RBX2 or ROC2, is a RING protein required for the activity of Cullin-RING ligase (CRL). Our recent study showed that Sag total knockout caused embryonic lethality at E11.5-12.5 days with associated defects in vasculogenesis. Whether Sag is required for de novo vasculogenesis in embryos and angiogenesis in tumors is totally unknown. Here, we report that Sag endothelial deletion also causes embryonic lethality at E15.5 with poor vasculogenesis. Sag deletion in primary endothelial cells (ECs) or knockdown in MS-1 ECs inhibits migration, proliferation and tube formation, with p27 accumulation being responsible for the suppression of migration and proliferation. Furthermore, Sag deletion significantly inhibits angiogenesis in an in vivo Matrigel plug assay, and tumor angiogenesis and tumorigenesis in a B16F10 melanoma model. Finally, MLN4924, an investigational small molecule inhibitor of NEDD8-activating enzyme (NAE) that inhibits CRL, suppresses in vitro migration, proliferation and tube formation, as well as in vivo angiogenesis and tumorigenesis. Taken together, our study, using both genetic and pharmaceutical approaches, demonstrates that Sag is essential for embryonic vasculogenesis and tumor angiogenesis, and provides the proof-of-concept evidence that targeting Sag E3 ubiquitin ligase may have clinical value for anti-angiogenesis therapy of human cancer. DOI: 10.1038/onc.2013.473 PMCID: PMC4016996 PMID: 24213570 [Indexed for MEDLINE] Conflict of interest statement: Conflicts of Interest The authors declare no conflict of interest.
http://www.ncbi.nlm.nih.gov/pubmed/18415121
1. Pflugers Arch. 2009 Jan;457(3):687-700. doi: 10.1007/s00424-008-0506-5. Epub 2008 Apr 16. The role of SERCA2a/PLN complex, Ca(2+) homeostasis, and anti-apoptotic proteins in determining cell fate. Vafiadaki E(1), Papalouka V, Arvanitis DA, Kranias EG, Sanoudou D. Author information: (1)Molecular Biology Division, Biomedical Research Foundation, Academy of Athens, Soranou Efesiou 4, Athens 115 27, Greece. Intracellular calcium is a major coordinator of numerous aspects of cellular physiology, including muscle contractility and cell survival. In cardiac muscle, aberrant Ca(2+) cycling has been implicated in a range of pathological conditions including cardiomyopathies and heart failure. The sarco(endo)plasmic reticulum Ca(2+) transport adenosine triphosphatase (SERCA2a) and its regulator phospholamban (PLN) have a central role in modulating Ca(2+) homeostasis and, therefore, cardiac function. Herein, we discuss the mechanisms through which SERCA2a and PLN control cardiomyocyte function in health and disease. Emphasis is placed on our newly identified PLN-binding partner HS-1-associated protein X-1 (HAX-1), which has an anti-apoptotic function and presents with numerous similarities to Bcl-2. Recent evidence indicates that proteins of the Bcl-2 family can influence ER Ca(2+) content, a critical determinant of cellular sensitivity to apoptosis. The discovery of the PLN/HAX-1 interaction therefore unveils an important new link between Ca(2+) homeostasis and cell survival, with significant therapeutic potential. DOI: 10.1007/s00424-008-0506-5 PMID: 18415121 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23624319
1. Anal Biochem. 2013 Aug 15;439(2):109-15. doi: 10.1016/j.ab.2013.04.016. Epub 2013 Apr 25. Quantifiable analysis of cellular pathway inhibition of a Nedd8-activating enzyme inhibitor, MLN4924, using AlphaScreen. Yan ZH(1), Burkhardt A, Loke HK, Chen J, Xu Q, Brauer P, Ma J, Lin Y, Garcia K, Dick LR, Bembenek ME. Author information: (1)Millennium Pharmaceuticals: The Takeda Oncology Company, Cambridge, MA 02139, USA. Cellular effects of a Nedd8-activating enzyme (NAE) inhibitor, MLN4924, using the AlphaScreen format were explored. MLN4924 acts as a substrate-assisted inhibitor of NAE by forming a tight binding Nedd8-MLN4924 adduct. The inhibited enzyme can no longer transfer Nedd8 downstream to modify and activate the E3 cullin-RING ligases. This results in the stabilization of proteins regulated by the proteasome, leading to cell death. These studies monitored the endogenous cellular changes to NAE∼Nedd8 thioester, the formation of the Nedd8-MLN4924 adduct, and the reduction in the Cul1-Nedd8. Lysates derived from MLN4924-treated HCT116 cells showed that whereas the β-subunit of NAE remained constant, reductions of both NAE∼Nedd8 thioester and Cul1-Nedd8 levels occurred with a concomitant rise of the adduct. Moreover, the formation of the Nedd8-MLN4924 adduct was approximately stoichiometric with the concentration of NAEβ. Higher density 384-well cell-based assays illustrated the kinetics of enzyme inactivation across a wider range of MLN4924 concentrations, showing a rapid loss of NAE∼Nedd8 thioester and Cul1-Nedd8. The reduction of NAE∼Nedd8 thioester precedes the loss of Cul1-Nedd8 at twice the rate. Finally, these results clearly demonstrate the utility of the homogeneous assay for quantitative assessment of these endogenous cellular components in a 384-well plate in response to inhibition of NAE by MLN4924. Copyright © 2013 Elsevier Inc. All rights reserved. DOI: 10.1016/j.ab.2013.04.016 PMID: 23624319 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17726868
1. Wiad Lek. 2007;60(3-4):155-7. [Sudden death in competitive athletes]. [Article in Polish] Juszczyk Z(1). Author information: (1)Szpitala im. Swietej Elzbiety w Białej. szpitalbiala@pro.onet.pl In athletes under the age of 35 years the incidence of sudden death is low, most causes to be due to ventricular arrhythmias, usually provoked by exertion, and nearly always occur in the presence of structural heart disease or abnormalities in the conduction system. The most common structural disease is hypertrophic cardiomyopathy followed by coronary artery anomalies, idiopathic dilated cardiomyopathy, arrhythmogenic right ventricular dysplasia, aortic stenosis, myocarditis, the Wolff-Parkinson-White syndrome, and long QT syndrome. The evaluation of athletes with symptoms of cardiac arrhythmias, syncope, family history of sudden death require a complete cardiac workup. If they have documented hypertrophic cardiomyopathy, arrhythmogenic right ventricular dysplasia, idiopathic dilated cardiomyopathy, long QT syndrome, family history presentation with sudden death, and septal thickness greater than 20 mm competitive athletics are generally prohibited. In athletes with asymptomatic bradyarrhythmia, supraventricular tachycardias and atrial premature contractions without structural heart disease all competitive sports are allowed if heart rate in bradyarrhythmia appropriately increases with exercise. Athletes with premature ventricular contraction, nonsustained ventricular tachycardia and non structural heart disease are without athletic restriction as long as the arrhythmia does not worsen on exertion and cause dyspnea, presyncope or syncope. PMID: 17726868 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16672832
1. Med Sci Sports Exerc. 2006 May;38(5):816-25. doi: 10.1249/01.mss.0000218130.41133.cc. The congenital long QT syndrome and implications for young athletes. Kapetanopoulos A(1), Kluger J, Maron BJ, Thompson PD. Author information: (1)Hartford Hospital, University of Connecticut School of Medicine, Farmington, 06102, USA. The congenital long QT syndrome (LQTS) is caused by cardiac ion channel mutations, which predispose young individuals to sudden cardiac death often related to exercise. The issue of LQTS and sports participation has received significant publicity due to reports of sudden death in young competitive athletes. This article reviews the pathophysiology, clinical characteristics, and management of LQTS in the physically active and athletic population. DOI: 10.1249/01.mss.0000218130.41133.cc PMID: 16672832 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24634471
1. Clin Cancer Res. 2014 Mar 15;20(6):1576-89. doi: 10.1158/1078-0432.CCR-13-0987. The Nedd8-activating enzyme inhibitor MLN4924 thwarts microenvironment-driven NF-κB activation and induces apoptosis in chronic lymphocytic leukemia B cells. Godbersen JC(1), Humphries LA, Danilova OV, Kebbekus PE, Brown JR, Eastman A, Danilov AV. Author information: (1)Authors' Affiliations: Departments of Medicine and Pathology, Dartmouth-Hitchcock Medical Center, Lebanon; Department of Pharmacology and Toxicology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire; and Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts. Erratum in Clin Cancer Res. 2016 Aug 15;22(16):4274. doi: 10.1158/1078-0432.CCR-16-1475. BACKGROUND: Stromal-mediated signaling enhances NF-κB pathway activity in chronic lymphocytic leukemia (CLL) B cells, leading to cell survival and chemoresistance. Ubiquitination of IκBα may partially account for constitutive activation of NF-κB. MLN4924 is an investigational agent that inhibits the Nedd8-activating enzyme, thereby neutralizing Cullin-RING ubiquitin ligases and preventing degradation of their substrates. EXPERIMENTAL DESIGN: We conducted a preclinical assessment of MLN4924 in CLL. Primary CLL cells were cocultured in vitro with CD40L-expressing stroma to mimic the prosurvival conditions present in lymphoid tissue. The effect of MLN4924 on CLL cell apoptosis, NF-κB pathway activity, Bcl-2 family members, and cell cycle was assessed by flow cytometry, Western blotting, PCR, and immunocytochemistry. RESULTS: CD40L-expressing stroma protected CLL cells from spontaneous apoptosis and induced resistance to multiple drugs, accompanied by NF-κB activation and Bim repression. Treatment with MLN4924 induced CLL cell apoptosis and circumvented stroma-mediated resistance. This was accompanied by accumulation of phospho-IκBα, decreased nuclear translocation of p65 and p52 leading to inhibition of both the canonical and noncanonical NF-κB pathways, and reduced transcription of their target genes, notably chemokines. MLN4924 promoted induction of Bim and Noxa in the CLL cells leading to rebalancing of Bcl-2 family members toward the proapoptotic BH3-only proteins. siRNA-mediated knockdown of Bim or Noxa decreased sensitivity to MLN4924. MLN4924 enhanced the antitumor activity of the inhibitors of B-cell receptor (BCR)-associated kinases. CONCLUSIONS: MLN4924 disrupts NF-κB activation and induces Bim expression in CLL cells, thereby preventing stroma-mediated resistance. Our data provide rationale for further evaluation of MLN4924 in CLL. ©2014 AACR. DOI: 10.1158/1078-0432.CCR-13-0987 PMCID: PMC3960291 PMID: 24634471 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24198575
1. Open Access J Sports Med. 2011 Jul 21;2:85-97. doi: 10.2147/OAJSM.S10675. Sudden cardiac death in young athletes. Ostman-Smith I(1). Author information: (1)Division of Paediatric Cardiology, Institute of Clinical Sciences, Sahlgrenska Academy, Gothenburg University, Sweden. Athletic activity is associated with an increased risk of sudden death for individuals with some congenital or acquired heart disorders. This review considers in particular the causes of death affecting athletes below 35 years of age. In this age group the largest proportion of deaths are caused by diseases with autosomal dominant inheritance such as hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, long QT-syndrome, and Marfan's syndrome. A policy of early cascade-screening of all first-degree relatives of patients with these disorders will therefore detect a substantial number of individuals at risk. A strictly regulated system with preparticipation screening of all athletes following a protocol pioneered in Italy, including school-age children, can also detect cases caused by sporadic new mutations and has been shown to reduce excess mortality among athletes substantially. Recommendations for screening procedure are reviewed. It is concluded that ECG screening ought to be part of preparticipation screening, but using criteria that do not cause too many false positives among athletes. One such suggested protocol will show positive in approximately 5% of screened individuals, among whom many will be screened for these diseases. On this point further research is needed to define what kind of false-positive and false-negative rate these new criteria result in. A less formal system based on cascade-screening of relatives, education of coaches about suspicious symptoms, and preparticipation questionnaires used by athletic clubs, has been associated over time with a sizeable reduction in sudden cardiac deaths among Swedish athletes, and thus appears to be worth implementing even for junior athletes not recommended for formal preparticipation screening. It is strongly argued that in families with autosomal dominant disorders the first screening of children should be carried out no later than 6 to 7 years of age. DOI: 10.2147/OAJSM.S10675 PMCID: PMC3781887 PMID: 24198575
http://www.ncbi.nlm.nih.gov/pubmed/19535269
1. Cardiovasc Pathol. 2010 Jul-Aug;19(4):207-17. doi: 10.1016/j.carpath.2009.04.001. Epub 2009 Jun 16. Prevention of sudden cardiac death in the young and in athletes: dream or reality? Thiene G(1), Carturan E, Corrado D, Basso C. Author information: (1)Pathology Department of Medical Diagnostic Sciences and Special Therapies, University of Padua Medical School, Padua, Italy. gaetano.thiene@unipd.it Cardiovascular diseases account for 40% of all deaths in the Western countries, and nearly two thirds of them occur suddenly. Young people (<35 years) are not spared from sudden death (SD) with a rate of 1/100,000 per year. Effort is a trigger with a threefold risk in athletes vs. nonathletes, and sports disqualification is by itself life-saving in people with underlying concealed cardiovascular diseases. Several culprits of cardiac SD may be identified at postmortem and atherosclerotic coronary artery disease is the leading cause (25% of SD cases in the young), mostly consisting of a single obstructive plaque with fibrocellular intimal proliferation. However, the spectrum of cardiovascular substrates is wide and include also congenital diseases of the coronary arteries (mainly anomalous origin), myocardium (arrhythmogenic and hypertrophic cardiomyopathies, myocarditis), valves (aortic stenosis and mitral valve prolapse), and conduction system (ventricular preexcitation, accelerated atrioventricular conduction and block). In up to 20% of cases, the heart is grossly and histologically normal at autopsy (unexplained SD or "mors sine materia"), and inherited ion channel diseases have been implicated (long and short QT syndromes, Brugada syndrome, catecholaminergic polymorphic ventricular tachycardia). Targets to treat and prevent SD in the young consist of the following: (a) avoid triggers like effort or emotion, (b) inhibit the onset of arrhythmias with drugs or ablation, (c) switch off arrhythmias with defibrillator, and (d) hinder the recurrence of the disease with genetic counseling and/or therapy. In vivo detection of cardiomyopathies is nowadays feasible by electrocardiogram and/or echocardiography, which resulted in a sharp decline of SD in the athletes in Italy, thanks to obligatory preparticipation screening for sport activity. Genetic screening could play a pivotal role in early detection of asymptomatic mutation carriers of cardiovascular diseases at risk of SD. DOI: 10.1016/j.carpath.2009.04.001 PMID: 19535269 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15074012
1. Pol Merkur Lekarski. 2004 Jan;16(91):5-7. [Cardiovascular diseases as a cause of sudden death in athletes]. [Article in Polish] Halawa B. Cardiac arrhythmias are the reason of the most sudden deaths in athletes. The annual risk of sudden death at athletes is between 5 to 10 per one million. Benign arrhythmia including bradyarrhythmias, atrial and ventricular premature contractions are common in the athletes. Supraventricular arrhythmias such as atrial fibrillation, nodal reciprocal entrant tachycardia and Wolff-Parkinson-White syndrome are less common. Perhaps the rarest and the most dangerous arrhythmias are ventricular arrhythmias, among them arrhythmias secondary to hypertrophic cardiomyopathy, arrhythmogenic right ventricular dysplasia, long QT syndrome, and anomalous origin of coronary arteries. Asymptomatic bradyarrhythmias (if the heart rate in bradyarrhythmia appropriate increases with exercise), supraventricularis tachycardias, and atrial premature contractions without structural heart disease are not the contraindication to sports Athletes with premature ventricular contraction, nonsustained ventricular tachycardia and non structural heart disease are without athletic restrictions as long as the arrhythmias do not worsen and they not cause dyspnea or presyncope during exertion. Frequent or multiform premature ventricular contraction or sustained ventricular tachycardia indicate a higher risk, and all participation in athletic should be restricted. PMID: 15074012 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23986575
1. J Virol. 2013 Nov;87(21):11741-50. doi: 10.1128/JVI.02002-13. Epub 2013 Aug 28. Inhibition of CUL4A Neddylation causes a reversible block to SAMHD1-mediated restriction of HIV-1. Hofmann H(1), Norton TD, Schultz ML, Polsky SB, Sunseri N, Landau NR. Author information: (1)Microbiology Department, New York University School of Medicine, New York, New York, USA. The deoxynucleoside triphosphohydrolase SAMHD1 restricts retroviral replication in myeloid cells. Human immunodeficiency virus type 2 (HIV-2) and a simian immunodeficiency virus from rhesus macaques (SIVmac) encode Vpx, a virion-packaged accessory protein that counteracts SAMHD1 by inducing its degradation. SAMHD1 is thought to work by depleting the pool of intracellular deoxynucleoside triphosphates but has also been reported to have exonuclease activity that could allow it to degrade the viral genomic RNA or viral reverse-transcribed DNA. To induce the degradation of SAMHD1, Vpx co-opts the cullin4a-based E3 ubiquitin ligase, CRL4. E3 ubiquitin ligases are regulated by the covalent attachment of the ubiquitin-like protein Nedd8 to the cullin subunit. Neddylation can be prevented by MLN4924, a drug that inhibits the nedd8-activating enzyme. We report that MLN4924 inhibits the neddylation of CRL4, blocking Vpx-induced degradation of SAMHD1 and maintaining the restriction. Removal of the drug several hours postinfection released the block. Similarly, Vpx-containing virus-like particles and deoxynucleosides added to the cells more than 24 h postinfection released the SAMHD1-mediated block. Taken together, these findings support deoxynucleoside triphosphate pool depletion as the primary mechanism of SAMHD1 restriction and argue against a nucleolytic mechanism, which would not be reversible. DOI: 10.1128/JVI.02002-13 PMCID: PMC3807335 PMID: 23986575 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15184297
1. Circulation. 2004 Jun 8;109(22):2807-16. doi: 10.1161/01.CIR.0000128363.85581.E1. Recommendations for physical activity and recreational sports participation for young patients with genetic cardiovascular diseases. Maron BJ, Chaitman BR, Ackerman MJ, Bayés de Luna A, Corrado D, Crosson JE, Deal BJ, Driscoll DJ, Estes NA 3rd, Araújo CG, Liang DH, Mitten MJ, Myerburg RJ, Pelliccia A, Thompson PD, Towbin JA, Van Camp SP; Working Groups of the American Heart Association Committee on Exercise, Cardiac Rehabilitation, and Prevention; Councils on Clinical Cardiology and Cardiovascular Disease in the Young. A group of relatively uncommon but important genetic cardiovascular diseases (GCVDs) are associated with increased risk for sudden cardiac death during exercise, including hypertrophic cardiomyopathy, long-QT syndrome, Marfan syndrome, and arrhythmogenic right ventricular cardiomyopathy. These conditions, characterized by diverse phenotypic expression and genetic substrates, account for a substantial proportion of unexpected and usually arrhythmia-based fatal events during adolescence and young adulthood. Guidelines are in place governing eligibility and disqualification criteria for competitive athletes with these GCVDs (eg, Bethesda Conference No. 26 and its update as Bethesda Conference No. 36 in 2005). However, similar systematic recommendations for the much larger population of patients with GCVD who are not trained athletes, but nevertheless wish to participate in any of a variety of recreational physical activities and sports, have not been available. The practicing clinician is frequently confronted with the dilemma of designing noncompetitive exercise programs for athletes with GCVD after disqualification from competition, as well as for those patients with such conditions who do not aspire to organized sports. Indeed, many asymptomatic (or mildly symptomatic) patients with GCVD desire a physically active lifestyle with participation in recreational and leisure-time activities to take advantage of the many documented benefits of exercise. However, to date, no reference document has been available for ascertaining which types of physical activity could be regarded as either prudent or inadvisable in these subgroups of patients. Therefore, given this clear and present need, this American Heart Association consensus document was constituted, based largely on the experience and insights of the expert panel, to offer recommendations governing recreational exercise for patients with known GCVDs. DOI: 10.1161/01.CIR.0000128363.85581.E1 PMID: 15184297 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20675977
1. Postgrad Med. 2010 Jul;122(4):144-57. doi: 10.3810/pgm.2010.07.2181. Causes of sudden cardiac arrest in young athletes. Westrol MS(1), Kapitanyan R, Marques-Baptista A, Merlin MA. Author information: (1)University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, NJ, USA. Knowledge of sudden cardiac death in young athletes is imperative for all physicians and allied health professionals. The complete differential diagnosis of a young patient with sudden cardiac arrest will result in proper work-up and treatment. In this article, we review several etiologies of sudden cardiac death, including hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, Wolff-Parkinson-White syndrome, long QT syndrome, Brugada syndrome, and commotio cordis. Clinical findings, work-up, treatment, long-term management, and athlete preparticipation screening guidelines are discussed. DOI: 10.3810/pgm.2010.07.2181 PMID: 20675977 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12831662
1. Curr Sports Med Rep. 2003 Apr;2(2):72-8. doi: 10.1249/00149619-200304000-00005. The long QT syndrome: considerations in the athletic population. Schulze-Bahr E(1), Mönnig G, Eckardt L, Wedekind H, Wichter T, Breithardt G. Author information: (1)Department Molekular-Kardiologie, Institut für Arterioskleroseforschung an der Westfälischen, Wilhelms-Universität Münster, Domagkstrasse 3, D-48149 Münster, Germany. heart@uni-muenster.de In athletes, ventricular arrhythmias and sudden cardiac death are rare and unpredictable events. Often, an underlying heart disease is present, but pre-existing clinical signs or symptoms may not be recognized. Primary electrical disorders (such as the long QT syndrome) are rarely present in athletes but, so far, are a considerable reason for disqualification from sport activity. These disorders are mostly inherited, and patients should be referred to a cardiologist with special experience. Through the efforts of molecular genetics and cellular electrophysiology, an increasing understanding of the underlying mechanisms of arrhythmogenesis is being gathered. During the past decade, evidence has grown that establishing accurate genetic diagnoses and dissection of molecular disease mechanisms can have an impact on prognosis, and help direct therapy in a range of cardiovascular diseases. Further achievements in the areas of clinical and molecular research, improvement of medical education, and expansion of genotyping facilities will facilitate the correct and immediate identification of affected patients. DOI: 10.1249/00149619-200304000-00005 PMID: 12831662 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11475927
1. Rev Med Liege. 2001 May;56(5):318-25. [Sudden death in athletes]. [Article in French] Kulbertus H(1). Author information: (1)Service de Cardiologie, Université de Liège. Sudden death is rare in the young athlete. The causes may vary. In the US, hypertrophic cardiomyopathy plays the predominant role whereas in Europe right ventricular arrhythmogenic dysplasia and atherosclerosis of the coronary arteries are more frequent. Other causes such as congenital anomalies of the coronary vessels, myocarditis, Marfan's disease, the long QT, the Brugada and the Wollf-Parkinson-White syndromes exist, but are rare. Attentive preparticipation screening (clinical history and medical examination) is mandatory in all future young athletes. PMID: 11475927 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20975745
1. Nat Rev Clin Oncol. 2010 Dec;7(12):725-32. doi: 10.1038/nrclinonc.2010.170. Epub 2010 Oct 26. Breast cancer assessment tools and optimizing adjuvant therapy. Oakman C(1), Santarpia L, Di Leo A. Author information: (1)Sandro Pitigliani Medical Oncology Unit, Department of Oncology, Hospital of Prato, Istituto Toscano Tumori, Piazza Ospedale 2, 59100 Prato, Italy. Recommendation of systemic adjuvant therapy and choice of optimal agents for early-stage breast cancer remains a challenge. Adjuvant therapy is indicated on the assumption of residual micrometastatic disease. Adjuvant assessment tools for prognosis and prediction of treatment benefit, including Adjuvant! Online, the St Gallen Consensus, Oncotype DX(®) and MammaPrint(®), aid clinical decision making. However, all of these tools have limitations that must be considered in their judicious application. Clinicopathological based tools are critically dependent on accurate, standardized measurement of parameters. Multigene tools are appealing for their objectivity and reproducibility, particularly regarding analysis of proliferation, but these approaches still overlook the biological heterogeneity within tumors evidenced by distinct cell subpopulations with different genomic patterns and function. The greatest treatment challenge remains for patients assessed as intermediate risk of relapse, a problem not overcome by multigene tools. Remarkable diversity in breast cancer dictates that adjuvant management must be biologically driven. Future identification of predictive biomarkers for specific chemotherapy sensitivity may allow targeted use of available agents, including anthracyclines, taxanes and DNA damaging agents. The presence of drug targets and targetable signaling pathways, rather than molecularly defined subgroups, may ultimately drive treatment decisions. DOI: 10.1038/nrclinonc.2010.170 PMID: 20975745 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18159035
1. Am J Health Syst Pharm. 2008 Jan 1;65(1):23-8. doi: 10.2146/ajhp060352. Gene-expression assays: new tools to individualize treatment of early-stage breast cancer. Dobbe E(1), Gurney K, Kiekow S, Lafferty JS, Kolesar JM. Author information: (1)School of Pharmacy, University of Wisconsin, Madison, WI 53705-2222, USA. PURPOSE: The clinical and economic data for the two currently available gene-expression assays are reviewed. SUMMARY: Two gene-expression assays, used to determine the risk of breast cancer recurrence in patients with stage I or II node-negative breast cancer, are currently available. Oncotype DX is an assay performed on RNA extracted from paraffin-embedded tumor tissue. It analyzes the expression of 21 genes: 16 cancer-related genes and 5 reference genes. The results are used to calculate a recurrence score to identify the likelihood of cancer recurrence in patients treated with tamoxifen. The results of two studies evaluating the ability of Oncotype DX to predict the risk of breast cancer recurrence suggest that patients with ER-positive, node-negative breast cancer and a low recurrence score may need only adjuvant treatment with tamoxifen, while intermediate- and high-risk patients may require additional treatment with adjuvant chemotherapy. MammaPrint, an oligonucleotide microassay performed on fresh-frozen tumor samples, analyzes the expression of 70 genes. Studies have found that MammaPrint allows young patients (<61 years) with early-stage breast cancer to be categorized as having a high or low risk of distant metastasis. High-risk patients may then be managed with more aggressive therapy. CONCLUSION: Two gene-expression assays, Oncotype DX and MammaPrint, have been developed to determine the risk of breast cancer recurrence in patients with stage I or II node-negative breast cancer. In the future, these tests may be useful in determining the need for systemic adjuvant therapy in such patients. DOI: 10.2146/ajhp060352 PMID: 18159035 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19825882
1. Ann Oncol. 2010 Apr;21(4):717-722. doi: 10.1093/annonc/mdp388. Epub 2009 Oct 13. The 70-gene prognosis signature predicts early metastasis in breast cancer patients between 55 and 70 years of age. Mook S(1), Schmidt MK(2), Weigelt B(3), Kreike B(4), Eekhout I(1), van de Vijver MJ(1), Glas AM(5), Floore A(5), Rutgers EJT(6), van 't Veer LJ(7). Author information: (1)Department of Pathology. (2)Department of Epidemiology. (3)Department of Experimental Therapy. (4)Department of Radiation Oncology, The Netherlands Cancer Institute. (5)Department of Agendia BV. (6)Department of Surgical Oncology, The Netherlands Cancer Institute, Amsterdam, The Netherlands. (7)Department of Pathology; Department of Agendia BV. Electronic address: l.vt.veer@nki.nl. BACKGROUND: The majority of breast cancer patients are postmenopausal women who are increasingly being offered adjuvant chemotherapy. Since the beneficial effect of chemotherapy in postmenopausal patients predominantly occurs in the first 5 years after diagnosis, a prognostic marker for early events can be of use for adjuvant treatment decision making. The aim of this study was to evaluate the prognostic value of the 70-gene prognosis signature for early events in postmenopausal patients. METHODS: Frozen tumor samples from 148 patients aged 55-70 years were selected (T1-2, N0) and classified by the 70-gene prognosis signature (MammaPrint) into good or poor prognosis. Eighteen percent received hormonal therapy. RESULTS: Breast cancer-specific survival (BCSS) at 5 years was 99% for the good-prognosis signature versus 80% for the poor-prognosis signature group (P = 0.036). The 70-gene prognosis signature was a significant and independent predictor of BCCS during the first 5 years of follow-up with an adjusted hazard ratio of 14.4 (95% confidence interval 1.7-122.2; P = 0.01) at 5 years. CONCLUSION: The 70-gene prognosis signature can accurately select postmenopausal patients at low risk of breast cancer-related death within 5 years of diagnosis and can be of clinical use in selecting postmenopausal women for adjuvant chemotherapy. DOI: 10.1093/annonc/mdp388 PMID: 19825882 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19214742
1. Breast Cancer Res Treat. 2010 Feb;119(3):551-8. doi: 10.1007/s10549-009-0333-1. Epub 2009 Feb 13. The 70-gene signature as a response predictor for neoadjuvant chemotherapy in breast cancer. Straver ME(1), Glas AM, Hannemann J, Wesseling J, van de Vijver MJ, Rutgers EJ, Vrancken Peeters MJ, van Tinteren H, Van't Veer LJ, Rodenhuis S. Author information: (1)Department of Surgical Oncology, The Netherlands Cancer Institute - Antoni van Leeuwenhoek Hospital, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. m.straver@nki.nl The 70-gene signature (MammaPrint) is a prognostic tool used to guide adjuvant treatment decisions. The aim of this study was to assess its value to predict chemosensitivity in the neoadjuvant setting. We obtained the 70-gene profile of stage II-III patients prior to neoadjuvant chemotherapy and classified the prognosis-signatures. Pathological complete remission (pCR) was used to measure chemosensitivity. Among 167 patients, 144 (86%) were having a poor and 23 (14%) a good prognosis-signature. None of the good prognosis-signature patients achieved a pCR (0/23), whereas 29/144 patients (20%) in the poor prognosis-signature group did (P = 0.015). All triple-negative tumors (n = 38) had a poor prognosis-signature. Within the non triple-negative subgroup, the response of the primary tumor remained associated with the classification of the prognosis-signature (P = 0.023). A pCR is unlikely to be achieved in tumors that have a good prognosis-signature. Tumors with a poor prognosis-signature are more sensitive to chemotherapy. DOI: 10.1007/s10549-009-0333-1 PMID: 19214742 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19125125
1. Genet Med. 2009 Jan;11(1):66-73. doi: 10.1097/GIM.0b013e3181928f56. Recommendations from the EGAPP Working Group: can tumor gene expression profiling improve outcomes in patients with breast cancer? Evaluation of Genomic Applications in Practice and Prevention (EGAPP) Working Group(1). Collaborators: Berg AO, Armstrong K, Botkin J, Calonge N, Haddow J, Hayes M, Kaye C, Phillips KA, Piper M, Richards CS, Scott JA, Strickland OL, Teutsch S. Author information: (1)egappinfo@egappreviews.org SUMMARY OF RECOMMENDATIONS: The EGAPP Working Group (EWG) found insufficient evidence to make a recommendation for or against the use of tumor gene expression profiles to improve outcomes in defined populations of women with breast cancer. For one test, the EWG found preliminary evidence of potential benefit of testing results to some women who face decisions about treatment options (reduced adverse events due to low risk women avoiding chemotherapy), but could not rule out the potential for harm for others (breast cancer recurrence that might have been prevented). The evidence is insufficient to assess the balance of benefits and harms of the proposed uses of the tests. The EWG encourages further development and evaluation of these technologies. RATIONALE: The measurement of gene expression in breast tumor tissue is proposed as a way to estimate the risk of distant disease recurrence in order to provide additional information beyond current clinicopathological risk stratification and to influence decisions about treatment in order to improve health outcomes. Based on their review of the EGAPP-commissioned evidence report, Impact of Gene Expression Profiling Tests on Breast Cancer Outcomes and other data summaries, the EWG found no direct evidence linking tumor gene expression profiling of women with breast cancer to improved outcomes, and inadequate evidence to construct an evidence chain. However, further evaluation on the clinical utility of some tests and management algorithms, including well-designed randomized controlled trials, is warranted. ANALYTIC VALIDITY: Some data on technical performance of assays were identified for MammaPrint and Oncotype DX, though estimates of analytic sensitivity and specificity could not be made. Published performance data on the laboratory developed Quest H:I Test were limited. Overall, the EWG found the evidence to be inadequate. CLINICAL VALIDITY: The EWG found adequate evidence regarding the association of the Oncotype DX Recurrence Score with disease recurrence and adequate evidence for response to chemotherapy. The EWG found adequate evidence to characterize the association of MammaPrint with future metastases, but inadequate evidence to assess the added value to standard risk stratification, and could not determine the population to which the test would best apply. The evidence was inadequate to characterize the clinical validity of the Quest H:I Test. CLINICAL UTILITY: The EWG found no evidence regarding the clinical utility of the MammaPrint and Quest H:I Ratio tests, and inadequate evidence regarding Oncotype DX. These technologies have potential for both benefit and harm. CONTEXTUAL ISSUES: The EWG reviewed economic studies that used modeling to predict potential effects of using gene profiling, and judged the evidence inadequate. DOI: 10.1097/GIM.0b013e3181928f56 PMCID: PMC2743614 PMID: 19125125 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21847392
1. J Breast Cancer. 2011 Mar;14(1):33-8. doi: 10.4048/jbc.2011.14.1.33. Epub 2011 Mar 31. The 70-gene prognostic signature for korean breast cancer patients. Na KY(1), Kim KS, Lee JE, Kim HJ, Yang JH, Ahn SH, Moon BI, Kim RM, Ko SM, Jung YS. Author information: (1)Department of Surgery, Ajou University School of Medicine, Suwon, Korea. PURPOSE: A 70-gene prognostic signature has prognostic value in patients with node-negative breast cancer in Europe. This diagnostic test known as "MammaPrint™ (70-gene prognostic signature)" was recently validated and implementation was feasible. Therefore, we assessed the 70-gene prognostic signature in Korean patients with breast cancer. We compared the risk predicted by the 70-gene prognostic signature with commonly used clinicopathological guidelines among Korean patients with breast cancer. We also analyzed the 70-gene prognostic signature and clinicopathological feature of the patients in comparison with a previous validation study. METHODS: Forty-eight eligible patients with breast cancer (clinical T1-2N0M0) were selected from four hospitals in Korea. Fresh tumor samples were analyzed with a customized microarray for the 70-gene prognostic signature. Concordance between the risk predicted by the 70-gene prognostic signature and risk predicted by commonly used clinicopathological guidelines (St. Gallen guidelines, National Institutes of Health [NIH] guideline, and Adjuvant! Online) was evaluated. RESULTS: Prognosis signatures were assessed in 36 patients. No significant differences were observed in the clinicopathological features of patients compared with previous studies. The 70-gene prognosis signature identified five (13.9%) patients with a low-risk prognosis signature and 31 (86.1%) patients with a high-risk prognosis signature. Clinical risk was concordant with the prognosis signature for 29 patients (80.6%) according to the St. Gallen guidelines; 30 patients (83.4%) according to the NIH guidelines; and 23 patients (63.8%) according to the Adjuvant! Online. Our results were different from previous validation studies in Europe with about a 40% low-risk prognosis and about a 60% high-risk prognosis. The high incidence in the high-risk group was consistent with data in Japan. CONCLUSION: The results of 70-gene prognostic signature of Korean patients with breast cancer were somewhat different from those identified in Europe. This difference should be studied as whether there is a gene disparity between Asians and Europeans. Further large-scale studies with a follow-up evaluation are required to assess whether the use of the 70-gene prognostic signature can predict the prognosis of Korean patients with breast cancer. DOI: 10.4048/jbc.2011.14.1.33 PMCID: PMC3148507 PMID: 21847392 Conflict of interest statement: The authors declare that they have no competing interests.
http://www.ncbi.nlm.nih.gov/pubmed/23679081
1. Int J Neurosci. 2013 Nov;123(11):802-9. doi: 10.3109/00207454.2013.803477. Epub 2013 May 23. Involvement of the dual-specificity phosphatase M3/6 in c-Jun N-terminal kinase inactivation following cerebral ischemia in the rat hippocampus. Huang Z(1), Liu Y, Zhu J, Wu H, Guo J. Author information: (1)1Laboratory Center for Basic Medical Sciences, Nanjing Medical University , Nanjing , P.R. China. The c-Jun N-terminal kinase (JNK) undergoes complete inactivation following the intense activation induced by cerebral ischemia and reperfusion in rat hippocampi. This study examines the molecular mechanism underlying JNK dephosphorylation and inactivation evoked by dual-specificity phosphates following cerebral ischemia. The results revealed upregulation of dual-specificity phosphatase M3/6 (DUSP8) activity at 4 h of reperfusion in rat hippocampi. This was accompanied by the dephosphorylation of JNK. The M3/6 inhibitor, anisomycin, was found to enhance JNK activity following postischemic reperfusion, suggesting that M3/6 is closely associated with JNK inactivation following cerebral ischemia. Cerebral ischemia also induced an increase in heat shock protein (HSP70) levels, which is involved in the upregulation of soluble cytoplasmic M3/6 levels. The inhibition of HSP70 using quercetin resulted in an elevation of JNK activity by decreasing the cytoplasmic solubility of M3/6. The findings of the current study suggest that M3/6 is implicated in the inactivation of JNK in response to cerebral ischemia, which requires the molecular chaperone HSP70 to facilitate the correction of folding defects. DOI: 10.3109/00207454.2013.803477 PMID: 23679081 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/8910287
1. J Biol Chem. 1996 Nov 1;271(44):27205-8. doi: 10.1074/jbc.271.44.27205. The dual specificity phosphatases M3/6 and MKP-3 are highly selective for inactivation of distinct mitogen-activated protein kinases. Muda M(1), Theodosiou A, Rodrigues N, Boschert U, Camps M, Gillieron C, Davies K, Ashworth A, Arkinstall S. Author information: (1)Geneva Biomedical Research Institute, Glaxo Wellcome Research and Development S. A., CH-1228 Plan-les-Ouates, Geneva, Switzerland. SA7182@GGR.CO.UK The mitogen-activated protein (MAP) kinase family includes extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase/stress-activated protein kinase (JNK/SAPK) and p38/RK/CSBP (p38) as structurally and functionally distinct enzyme classes. Here we describe two new dual specificity phosphatases of the CL100/MKP-1 family that are selective for inactivating ERK or JNK/SAPK and p38 MAP kinases when expressed in COS-7 cells. M3/6 is the first phosphatase of this family to display highly specific inactivation of JNK/SAPK and p38 MAP kinases. Although stress-induced activation of p54 SAPKbeta, p46 SAPKgamma (JNK1) or p38 MAP kinases is abolished upon co-transfection with increasing amounts of M3/6 plasmid, epidermal growth factor-stimulated ERK1 is remarkably insensitive even to the highest levels of M3/6 expression obtained. In contrast to M3/6, the dual specificity phosphatase MKP-3 is selective for inactivation of ERK family MAP kinases. Low level expression of MKP-3 blocks totally epidermal growth factor-stimulated ERK1, whereas stress-induced activation of p54 SAPKbeta and p38 MAP kinases is inhibited only partially under identical conditions. Selective regulation by M3/6 and MKP-3 was also observed upon chronic MAP kinase activation by constitutive p21(ras) GTPases. Hence, although M3/6 expression effectively blocked p54 SAPKbeta activation by p21(rac) (G12V), ERK1 activated by p21(ras) (G12V) was insensitive to this phosphatase. ERK1 activation by oncogenic p21(ras) was, however, blocked totally by co-expression of MKP-3. This is the first report demonstrating reciprocally selective inhibition of different MAP kinases by two distinct dual specificity phosphatases. DOI: 10.1074/jbc.271.44.27205 PMID: 8910287 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11566103
1. Curr Biol. 2001 Sep 18;11(18):1439-43. doi: 10.1016/s0960-9822(01)00426-2. The JNK phosphatase M3/6 is inhibited by protein-damaging stress. Palacios C(1), Collins MK, Perkins GR. Author information: (1)The Department of Immunology, University College London Medical School, Windeyer Building, 46 Cleveland Street, London W1P6DB, United Kingdom. Cells respond to stresses such as osmotic shock and heat shock by activating stress-activated protein kinases (SAPKs), including c-Jun N-terminal kinase (JNK) [1]. Activation of JNK requires phosphorylation of threonine and tyrosine residues in the TPY activation loop motif [2, 3] and can be reversed by the removal of either phosphate group. Numerous JNK phosphatases including dual-specificity phosphatases [4, 5], have been identified. Many stimuli activate JNK by increasing its rate of phosphorylation; however, JNK dephosphorylation is inhibited in cells after heat shock [6], suggesting that a JNK phosphatase(s) is inactivated. M3/6 is a dual-specificity phosphatase selective for JNK [7, 8]. We have previously expressed M3/6 in the mouse bone marrow cell line BAF3 in order to show that JNK activation by IL-3 is necessary for cell survival and proliferation [9]. Here we report that M3/6 dissociates from JNK and appears in an insoluble fraction after heat shock. These data identify M3/6 as a JNK phosphatase that is inactivated by heat shock and provide a molecular mechanism for the activation of JNK by heat shock. DOI: 10.1016/s0960-9822(01)00426-2 PMID: 11566103 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22100391
1. Cell Signal. 2012 Mar;24(3):664-76. doi: 10.1016/j.cellsig.2011.10.015. Epub 2011 Nov 9. Phosphorylation of the M3/6 dual-specificity phosphatase enhances the activation of JNK by arsenite. Cotsiki M(1), Oehrl W, Samiotaki M, Theodosiou A, Panayotou G. Author information: (1)Institute of Molecular Oncology, Biomedical Sciences Research Center Alexander Fleming, Vari, Greece. Specific outcomes upon activation of the c-Jun N-terminal kinase (JNK) pathway critically depend on the intensity and duration of signal transmission. Dual-specificity phosphatases (DUSPs) play a very important role in these events by modulating the extent of JNK phosphorylation and activation and thus regulating cellular responses to stress. M3/6 (DUSP8) is one of the dual-specificity protein phosphatases with distinct specificity towards JNK. It has been shown that M3/6 itself is phosphorylated by JNK upon stimulation with arsenite, but the role of this phosphorylation has not been investigated. In this study, we mapped JNK-induced phosphorylation sites on M3/6 using mass spectrometry. Phosphorylated residues Ser 515, Thr 518 and Ser 520 were identified and site-directed mutagenesis was employed to investigate their role. Upon arsenite stimulation, M3/6 mutated at these sites exhibited decreased phosphorylation compared to the wild-type protein. No difference was observed in terms of the enzyme's in vitro phosphatase activity, its substrate specificity towards JNK isoforms, its interactions with JNK and the scaffold family of JNK-interacting proteins (JIPs), its stability or its subcellular localization. Interestingly, expression of M3/6 phosphorylation mutants delayed the time-course of JNK phosphorylation and activation by arsenite. We propose that phosphorylation of the M3/6 phosphatase by JNK in response to stress stimuli results in attenuation of phosphatase activity and acceleration of JNK activation. Copyright © 2011 Elsevier Inc. All rights reserved. DOI: 10.1016/j.cellsig.2011.10.015 PMID: 22100391 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12598532
1. J Biol Chem. 2003 May 9;278(19):16957-67. doi: 10.1074/jbc.M212049200. Epub 2003 Feb 21. Polyglutamine expansion induces a protein-damaging stress connecting heat shock protein 70 to the JNK pathway. Merienne K(1), Helmlinger D, Perkin GR, Devys D, Trottier Y. Author information: (1)Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS, INSERM, Université Louis Pasteur, BP 10142, Illkirch C U de Strasbourg, 67404 cedex, France. merienne@igbmc.u-strasbg.fr Polyglutamine diseases, including Huntington's disease, designate a group of nine neurodegenerative disorders characterized by the presence of a toxic polyglutamine expansion in specific target proteins. Using cell and mouse models, we have shown that expanded polyglutamine led to activation of the stress kinase JNK and the transcription factor AP-1, which are implicated in neuronal death. Polyglutamine expansion-induced stress shared common features with protein-damaging stress such as heat shock, because activation of JNK involved inhibition of JNK phosphatase activities. Indeed, expanded polyglutamine impaired the solubility of the dual-specificity JNK phosphatase M3/6. Aggregation of M3/6 by polyglutamine expansion appeared to be indirect, because M3/6 was not recruited into polyglutamine inclusions. The heat shock protein HSP70, which is known to inhibit JNK during the heat shock response, suppressed polyglutamine-mediated aggregation of M3/6 and activation of JNK. Interestingly, levels of HSP70 were down-regulated by polyglutamine expansion. We suggest that reduction of HSP70 by expanded polyglutamine is implicated in aggregation and inhibition of M3/6 and in activation of JNK and AP-1. DOI: 10.1074/jbc.M212049200 PMID: 12598532 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11948422
1. Oncogene. 2002 Apr 4;21(15):2387-97. doi: 10.1038/sj.onc.1205309. Differential effects of stress stimuli on a JNK-inactivating phosphatase. Theodosiou A(1), Ashworth A. Author information: (1)CRC Gene Function and Regulation Group, The Breakthrough Toby Robins Breast Cancer Research Centre, Institute of Cancer Research, Fulham Road, London SW3 6JB, UK. Stress signals elicit a wide variety of cellular responses, many of which converge on the phosphorylation of JNK and p38 kinases, the activation of which has been well-characterized. How these kinases are switched off by dephosphorylation is not well understood. Here we describe how diverse cellular stresses affect differently the stability and activity of a JNK-inactivating dual-specificity threonine-tyrosine phosphatase M3/6. Both anisomycin and arsenite activate the JNK pathway and, in addition, inactivate the M3/6 phosphatase. However, while anisomycin treatment of cells leads to M3/6 protein degradation, arsenite appears to inactivate M3/6 directly. These results might have implications for the mechanism of tumour promotion by arsenic. DOI: 10.1038/sj.onc.1205309 PMID: 11948422 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21234187
1. Ann Pediatr Cardiol. 2010 Jul;3(2):107-12. doi: 10.4103/0974-2069.74035. Sudden cardiac death in children and adolescents (excluding Sudden Infant Death Syndrome). Gajewski KK(1), Saul JP. Author information: (1)Department of Pediatrics, Louisiana State University School of Medicine, New Orleans, Louisiana, USA. Sudden death in the young is rare. About 25% of cases occur during sports. Most young people with sudden cardiac death (SCD) have underlying heart disease, with hypertrophic cardiomyopathy and coronary artery anomalies being commonest in most series. Arrhythmogenic right ventricular dysplasia and long QT syndrome are the most common primary arrhythmic causes of SCD. It is estimated that early cardiopulmonary resuscitation and widespread availability of automatic external defibrillators could prevent about a quarter of pediatric sudden deaths. DOI: 10.4103/0974-2069.74035 PMCID: PMC3017912 PMID: 21234187 Conflict of interest statement: Conflict of Interest: None declared.
http://www.ncbi.nlm.nih.gov/pubmed/10915787
1. J Biol Chem. 2000 Oct 13;275(41):31755-62. doi: 10.1074/jbc.M004182200. Regulation of dual-specificity phosphatases M3/6 and hVH5 by phorbol esters. Analysis of a delta-like domain. Johnson TR(1), Biggs JR, Winbourn SE, Kraft AS. Author information: (1)Department of Medical Oncology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA. Treatment of leukemic cells with phorbol 12-myristate 13-acetate (PMA) induces a short-lived phosphorylation and activation of stress-activated protein kinase (SAPK) and cellular differentiation. To investigate whether the rapid deactivation of SAPK results from dephosphorylation by dual-specificity phosphatases (DSPs), we studied regulation of the DSP hVH5 and its murine orthologue M3/6 in K562 human leukemia cells. PMA treatment rapidly induced hVH5 transcripts in these cells, and induced expression of M3/6 completely inhibited PMA-stimulated phosphorylation of SAPK, suggesting a feedback loop to control SAPK activity. Using both stable cell lines and transient transfection we demonstrate that activation of SAPK rapidly stimulated phosphorylation of M3/6. This phosphorylation did not regulate the half-life of total cellular M3/6. hVH5 and M3/6 shares with all sequenced mammalian DSPs an amino acid motif, XILPXLXL, located approximately 80 amino acids from the active site. The hVH5-M3/6 sequence, RILPHLYL, shares significant homology with the SAPK binding site of the c-Jun protein, called the delta domain. This motif was found to be important for DSP function, because deletion of RILPHLYL inhibits SAPK-mediated phosphorylation of M3/6, and deletion of this sequence or mutation of the LYL portion blocks the ability of this phosphatase to dephosphorylate SAPK. DOI: 10.1074/jbc.M004182200 PMID: 10915787 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15888437
1. J Biol Chem. 2005 Jul 8;280(27):25651-8. doi: 10.1074/jbc.M501926200. Epub 2005 May 10. Dynamic interaction between the dual specificity phosphatase MKP7 and the JNK3 scaffold protein beta-arrestin 2. Willoughby EA(1), Collins MK. Author information: (1)Division of Infection and Immunity, University College London, Windeyer Building, 46 Cleveland Street, London W1T 4JF, United Kingdom. JNK scaffold proteins bind JNK and upstream kinases to activate subsets of JNK and localize activated JNK to specific subcellular sites. We previously demonstrated that the dual specificity phosphatases (DSPs) MKP7 and M3/6 bind the scaffold JNK-interacting protein-1 (JIP-1) and inactivate the bound subset of JNK (1). The G protein-coupled receptor (GPCR) adaptor beta-arrestin 2 is also a JNK3 scaffold. It binds the upstream kinases ASK1 and MKK4 and couples stimulation of the angiotensin II receptor AT1aR to activation of a cytoplasmic pool of JNK3. Here we report that MKP7 also binds beta-arrestin 2 via amino acids 394-443 of MKP7, the same region that interacts with JIP-1. This region of MKP7 interacts with beta-arrestin 2 at a central region near the JNK binding domain. MKP7 dephosphorylates JNK3 bound to beta-arrestin 2, either following activation by ASK1 overexpression or following AT1aR stimulation. Initial AT1aR stimulation causes a rapid (within 5 min) dissociation of MKP7 from beta-arrestin 2. MKP7 then reassociates with beta-arrestin 2 on endocytic vesicles 30-60 min after initial receptor stimulation. This dynamic interaction between phosphatase and scaffold permits signal transduction through a module that binds both positive and negative regulators. DOI: 10.1074/jbc.M501926200 PMID: 15888437 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23159405
1. Cell Signal. 2013 Feb;25(2):429-38. doi: 10.1016/j.cellsig.2012.11.010. Epub 2012 Nov 15. Differential regulation of M3/6 (DUSP8) signaling complexes in response to arsenite-induced oxidative stress. Oehrl W(1), Cotsiki M, Panayotou G. Author information: (1)Biomedical Sciences Research Center "Alexander Fleming", Vari 166 72, Greece. Mitogen-activated protein kinase (MAPK) cascades are involved in the regulation of cellular proliferation, differentiation, survival, apoptosis, as well as in inflammatory responses. Signal intensity and duration have been recognized as crucial parameters determining MAPK signaling output. Phosphatases play a particularly important role in this respect, by tightly controlling MAPK phosphorylation and activation. M3/6 (DUSP8) is a dual-specificity phosphatase implicated in the dephosphorylation and inactivation of JNK and, to a lesser extent, p38 MAPKs and is found in a complex with these kinases, along with other pathway components, held together by scaffold proteins. The JNK family consists of three genes, giving rise to at least ten different splice variants. Some functional differences between these gene products have been demonstrated, but the underlying molecular mechanisms and the roles of individual splice variants are still incompletely understood. We have investigated the interaction of M3/6 with JNK isoforms, as well as scaffold proteins of the JNK interacting protein (JIP) family, in order to elucidate the contribution of M3/6 to the regulation of distinct JNK signaling modules. M3/6 exhibited stronger binding towards JNK1β and JNK2α isoforms and this was reflected in higher enzymatic activity towards JNK2α2 when compared to JNK1α1 in vitro. After activation of the pathway by exposure of cells to arsenite, the interaction of M3/6 with JNK1α and JNK3 was enhanced, whereas that with JNK1β or JNK2α decreased. The modulation of binding affinities was found to be independent of JNK-mediated M3/6 phosphorylation. Furthermore, arsenite treatment resulted in an inducible recruitment of M3/6 to JNK-interacting protein 3 (JIP3) scaffold complexes, while its interaction with JIP1 or JIP2 was constitutive. The presented data suggest an isoform-specific role for the M3/6 phosphatase and the dynamic targeting of M3/6 towards distinct JNK-containing signaling complexes. Copyright © 2012 Elsevier Inc. All rights reserved. DOI: 10.1016/j.cellsig.2012.11.010 PMID: 23159405 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20616184
1. Mol Cell Proteomics. 2010 Nov;9(11):2424-37. doi: 10.1074/mcp.M110.001917. Epub 2010 Jul 8. Naturally occurring human urinary peptides for use in diagnosis of chronic kidney disease. Good DM(1), Zürbig P, Argilés A, Bauer HW, Behrens G, Coon JJ, Dakna M, Decramer S, Delles C, Dominiczak AF, Ehrich JH, Eitner F, Fliser D, Frommberger M, Ganser A, Girolami MA, Golovko I, Gwinner W, Haubitz M, Herget-Rosenthal S, Jankowski J, Jahn H, Jerums G, Julian BA, Kellmann M, Kliem V, Kolch W, Krolewski AS, Luppi M, Massy Z, Melter M, Neusüss C, Novak J, Peter K, Rossing K, Rupprecht H, Schanstra JP, Schiffer E, Stolzenburg JU, Tarnow L, Theodorescu D, Thongboonkerd V, Vanholder R, Weissinger EM, Mischak H, Schmitt-Kopplin P. Author information: (1)Department of Chemistry, University of Wisconsin, Madison, Wisconsin 53706, USA. Because of its availability, ease of collection, and correlation with physiology and pathology, urine is an attractive source for clinical proteomics/peptidomics. However, the lack of comparable data sets from large cohorts has greatly hindered the development of clinical proteomics. Here, we report the establishment of a reproducible, high resolution method for peptidome analysis of naturally occurring human urinary peptides and proteins, ranging from 800 to 17,000 Da, using samples from 3,600 individuals analyzed by capillary electrophoresis coupled to MS. All processed data were deposited in an Structured Query Language (SQL) database. This database currently contains 5,010 relevant unique urinary peptides that serve as a pool of potential classifiers for diagnosis and monitoring of various diseases. As an example, by using this source of information, we were able to define urinary peptide biomarkers for chronic kidney diseases, allowing diagnosis of these diseases with high accuracy. Application of the chronic kidney disease-specific biomarker set to an independent test cohort in the subsequent replication phase resulted in 85.5% sensitivity and 100% specificity. These results indicate the potential usefulness of capillary electrophoresis coupled to MS for clinical applications in the analysis of naturally occurring urinary peptides. DOI: 10.1074/mcp.M110.001917 PMCID: PMC2984241 PMID: 20616184 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12524447
1. J Biol Chem. 2003 Mar 21;278(12):10731-6. doi: 10.1074/jbc.M207324200. Epub 2003 Jan 10. The JNK-interacting protein-1 scaffold protein targets MAPK phosphatase-7 to dephosphorylate JNK. Willoughby EA(1), Perkins GR, Collins MK, Whitmarsh AJ. Author information: (1)Department of Immunology and Molecular Pathology, University College London and Royal Free Medical School, Windeyer Institute, United Kingdom. The c-Jun N-terminal kinase (JNK) group of mitogen-activated protein kinases (MAPKs) are activated by pleiotropic signals including environmental stresses, growth factors, and hormones. A subset of JNK can bind to distinct scaffold proteins that also bind upstream kinases of the JNK pathway, allowing sequential kinase activation within a signaling module. The JNK-interacting protein-1 (JIP-1) scaffold protein specifically binds JNK, MAP kinase kinase 7, and members of the MLK family and is essential for stress-mediated JNK activation in neurones. Here we report that JIP-1 also binds the dual-specificity phosphatases MKP7 and M3/6 via a region independent of its JNK binding domain. The C-terminal region of MKP7, homologous to that of M3/6 but not other DSPs, is required for interaction with JIP-1. When MKP7 is bound to JIP-1 it reduces JNK activation leading to reduced phosphorylation of the JNK target c-Jun. These results indicate that the JIP-1 scaffold protein modulates JNK signaling via association with both protein kinases and protein phosphatases that target JNK. DOI: 10.1074/jbc.M207324200 PMID: 12524447 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21143788
1. BMC Genomics. 2010 Dec 1;11 Suppl 3(Suppl 3):S6. doi: 10.1186/1471-2164-11-S3-S6. Cross-domain neurobiology data integration and exploration. Xuan W(1), Dai M, Buckner J, Mirel B, Song J, Athey B, Watson SJ, Meng F. Author information: (1)Psychiatry Department and Molecular and Behavioral Neuroscience Institute, University of Michigan, USA. wxuan@umich.edu BACKGROUND: Understanding the biomedical implications of data from high throughput experiments requires solutions for effective cross-scale and cross-domain data exploration. However, existing solutions do not provide sufficient support for linking molecular level data to neuroanatomical structures, which is critical for understanding high level neurobiological functions. RESULTS: Our work integrates molecular level data with high level biological functions and we present results using anatomical structure as a scaffold. Our solution also allows the sharing of intermediate data exploration results with other web applications, greatly increasing the power of cross-domain data exploration and mining. CONCLUSIONS: The Flex-based PubAnatomy web application we developed enables highly interactive visual exploration of literature and experimental data for understanding the relationships between molecular level changes, pathways, brain circuits and pathophysiological processes. The prototype of PubAnatomy is freely accessible at: [http://brainarray.mbni.med.umich.edu/Brainarray/prototype/PubAnatomy]. DOI: 10.1186/1471-2164-11-S3-S6 PMCID: PMC2999351 PMID: 21143788 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19604367
1. BMC Genomics. 2009 Jul 15;10:316. doi: 10.1186/1471-2164-10-316. Genomic resources of Magnaporthe oryzae (GROMO): a comprehensive and integrated database on rice blast fungus. Thakur S(1), Jha S, Roy-Barman S, Chattoo B. Author information: (1)Centre for Genome Research, Department of Microbiology and Biotechnology Centre, Faculty of Science, The M, S, University of Baroda, Vadodara - 390002, India. bharat.chattoo@bcmsu.ac.in BACKGROUND: Magnaporthe oryzae, rice blast fungus, is the most devastating pathogen of rice. It has emerged as a model phytopathogen for the study of host-pathogen interactions. A large body of data has been generated on different aspects of biology of this fungus and on host-pathogen interactions. However, most of the data is scattered and is not available as a single resource for researchers in this field. DESCRIPTION: Genomic Resources of Magnaporthe oyzae (GROMO), is a specialized, and comprehensive database for rice blast fungus, integrating information from several resources. GROMO contains information on genomic sequence, mutants available, gene expression, localization of proteins obtained from a variety of repositories, as primary data. In addition, prediction of domains, pathways, protein-protein interactions, sumolyation sites and biochemical properties that were obtained after computational analysis of protein sequences have also been included as derived data. This database has an intuitive user interface that shall prompt the user to explore various possible information resources available on a given gene or a protein, from a single source. CONCLUSION: Currently, information on M. oryzae is available from different resources like BROAD MIT Magnaporthe database, Agrobacterium tumefaciens-mediated transformation (ATMT) M. oryzae database, Magnaporthe grisea--Oryza sativa (MGOS) and Massive Parallel Signature Sequencing (MPSS) databases. In the GROMO project, an effort has been made to integrate information from all these databases, derive some new data based on the available information analyzed by relevant programs and make more insightful predictions to better understand the biology of M. oryzae. The database is currently available at: http://gromo.msubiotech.ac.in/ DOI: 10.1186/1471-2164-10-316 PMCID: PMC2721851 PMID: 19604367 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23593176
1. PLoS One. 2013 Apr 4;8(4):e60213. doi: 10.1371/journal.pone.0060213. Print 2013. Microarray analyses of glucocorticoid and vitamin D3 target genes in differentiating cultured human podocytes. Cheng X(1), Zhao X, Khurana S, Bruggeman LA, Kao HY. Author information: (1)Department of Biochemistry, School of Medicine, Case Western Reserve University (CWRU) and the Comprehensive Cancer Center of CWRU, Cleveland, Ohio, United States of America. Erratum in PLoS One. 2013;8(6). doi:10.1371/annotation/ce9027f6-dbf9-4fc9-af40-9e244edf6a6e. Glomerular podocytes are highly differentiated epithelial cells that are key components of the kidney filtration units. Podocyte damage or loss is the hallmark of nephritic diseases characterized by severe proteinuria. Recent studies implicate that hormones including glucocorticoids (ligand for glucocorticoid receptor) and vitamin D3 (ligand for vitamin D receptor) protect or promote repair of podocytes from injury. In order to elucidate the mechanisms underlying hormone-mediated podocyte-protecting activity from injury, we carried out microarray gene expression studies to identify the target genes and corresponding pathways in response to these hormones during podocyte differentiation. We used immortalized human cultured podocytes (HPCs) as a model system and carried out in vitro differentiation assays followed by dexamethasone (Dex) or vitamin D3 (VD3) treatment. Upon the induction of differentiation, multiple functional categories including cell cycle, organelle dynamics, mitochondrion, apoptosis and cytoskeleton organization were among the most significantly affected. Interestingly, while Dex and VD3 are capable of protecting podocytes from injury, they only share limited target genes and affected pathways. Compared to VD3 treatment, Dex had a broader and greater impact on gene expression profiles. In-depth analyses of Dex altered genes indicate that Dex crosstalks with a broad spectrum of signaling pathways, of which inflammatory responses, cell migration, angiogenesis, NF-κB and TGFβ pathways are predominantly altered. Together, our study provides new information and identifies several new avenues for future investigation of hormone signaling in podocytes. DOI: 10.1371/journal.pone.0060213 PMCID: PMC3617172 PMID: 23593176 [Indexed for MEDLINE] Conflict of interest statement: Competing Interests: The authors have declared that no competing interests exist.
http://www.ncbi.nlm.nih.gov/pubmed/21044771
1. Adv Chronic Kidney Dis. 2010 Nov;17(6):487-92. doi: 10.1053/j.ackd.2010.09.005. Proteomics database in chronic kidney disease. Yamamoto T(1). Author information: (1)Institute of Nephrology, Structural Pathology, Chuo-ku Niigata, Niigata, Japan. tdsymmt@med.niigata-u.ac.jp Databases which are useful for proteomic analysis of human kidney tissue and urine have been discussed in this article. Integration of the gene-centric and protein-centric general databases with those of human kidney tissue and urine proteomes may open a new window for research in nephrology. Proteins present in the kidney and urine provide basic tools for investigation of kidney function and disease. By comparing such databases between the healthy and diseased populations, we may be able to identify the following: proteins involved in the development of renal disease, proteins involved in progression of CKD, or new biomarker candidate proteins for either the development of renal disease or the progression of CKD. Copyright © 2010 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved. DOI: 10.1053/j.ackd.2010.09.005 PMID: 21044771 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19728071
1. Behav Genet. 2009 Nov;39(6):605-15. doi: 10.1007/s10519-009-9294-8. Epub 2009 Sep 3. Born to be happy? The etiology of subjective well-being. Bartels M(1), Boomsma DI. Author information: (1)Department of Biological Psychology, VU University, Van der Boechorststraat 1, 1081 BT, Amsterdam, The Netherlands. m.bartels@psy.vu.nl Subjective Wellbeing (SWB) can be assessed with distinct measures that have been hypothesized to represent different domains of SWB. The current study assessed SWB with four different measures in a genetically informative sample of adolescent twins and their siblings aged 13-28 years (N = 5,024 subjects from 2,157 families). Multivariate genetic modeling was applied to the data to explore the etiology of individual differences in SWB measures and the association among them. Developmental trends and sex differences were examined for mean levels and the variance-covariance structure. Mean SWB levels were equal in men and women. A small negative effect of age on mean levels of SWB was found. Individual differences in SWB were accounted for by additive and non-additive genetic influences, and non-shared environment. The broad-sense heritabilities were estimated between 40 and 50%. The clustering of the four different measures (quality of life in general, satisfaction with life, quality of life at present, and subjective happiness) was explained by an underlying additive genetic factor and an underlying non-additive genetic factor. The effect of these latent genetic factors on the phenotypes was not moderated by either age or sex. DOI: 10.1007/s10519-009-9294-8 PMCID: PMC2780680 PMID: 19728071 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/2687698
1. Nature. 1989 Dec 21-28;342(6252):877-83. doi: 10.1038/342877a0. Conformations of immunoglobulin hypervariable regions. Chothia C(1), Lesk AM, Tramontano A, Levitt M, Smith-Gill SJ, Air G, Sheriff S, Padlan EA, Davies D, Tulip WR, et al. Author information: (1)MRC Laboratory of Molecular Biology, Cambridge, UK. Comment in Nature. 1990 Feb 1;343(6257):411-2. doi: 10.1038/343411a0. On the basis of comparative studies of known antibody structures and sequences it has been argued that there is a small repertoire of main-chain conformations for at least five of the six hypervariable regions of antibodies, and that the particular conformation adopted is determined by a few key conserved residues. These hypotheses are now supported by reasonably successful predictions of the structures of most hypervariable regions of various antibodies, as revealed by comparison with their subsequently determined structures. DOI: 10.1038/342877a0 PMID: 2687698 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/8227075
1. J Biol Chem. 1993 Nov 25;268(33):25124-31. Identification of a secondary Fc gamma RI binding site within a genetically engineered human IgG antibody. Chappel MS(1), Isenman DE, Oomen R, Xu YY, Klein MH. Author information: (1)Department of Immunology, University of Toronto, Ontario, Canada. Although human IgG2 is not cytophilic, we have shown previously that an IgG2 antibody expressing the sequence PLLGG (underline = substitution) spanning CH2 domain residues 233-237 (Eu numbering) displayed IgG1-like Fc gamma RI binding activity. In contrast, IgG1 PLLGG exhibited 3-fold less affinity, whereas IgG2 ELLGG was 3-fold more active than native IgG1. These results suggested that additional site(s) conferred enhanced binding properties to the engineered, cytophilic IgG2 variant. These sites were shown to reside in the IgG2 CH2 domain, since the IgG1 CH2 module did not have enhanced activity in a panel of hybrid IgG1/IgG2 antibodies. To map these sites further, human IgG1 and IgG2 constant region gene segments were modified to allow reciprocal COOH-terminal half segment exchanges of CH2 exons. These were cloned into a pSV2neo expression vector bearing a rearranged MOPC 315 heavy chain variable region gene and transfected into a MOPC 315 heavy chain deletion mutant. The dinitrophenol affinity-purified IgGs were radiolabeled and assessed for Fc gamma RI binding activity in direct binding assays using U937 cells. The COOH terminus of the IgG2 CH2 domain was found to contain accessory site(s) since it enhanced the binding properties of both IgG1 PLLGG and native IgG1. In contrast, grafting of the COOH terminus of the IgG1 CH2 domain onto IgG2 PLLGG and IgG2 ELLGG diminished their cytophilic activity. The amino acid responsible for the enhancing properties of the COOH terminus of the IgG2 CH2 domain was shown to be threonine 339, since IgG1 PLLGG/Thr339 displayed increased Fc gamma RI binding affinity. Kinetics studies revealed that this is accomplished through an increase in the forward rate constant of the IgG-Fc gamma RI interaction. PMID: 8227075 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22665257
1. Methods Mol Biol. 2012;882:605-33. doi: 10.1007/978-1-61779-842-9_33. IMGT/DomainGapAlign: the IMGT® tool for the analysis of IG, TR, MH, IgSF, and MhSF domain amino acid polymorphism. Ehrenmann F(1), Lefranc MP. Author information: (1)IMGT®, The International ImMunoGeneTics Information System®, Université Montpellier 2, Laboratoire d'ImmunoGénétique Moléculaire (LIGM), Institut de Génétique Humaine (IGH), UPR CNRS 1142, Montpellier, France. IMGT/DomainGapAlign is the online tool of IMGT(®), the international ImMunoGeneTics information system(®), for the analysis of amino acid sequences and two-dimensional (2D) structures of domains. IMGT/DomainGapAlign allows the analysis of the closest variable (V) and constant (C) domains of immunoglobulins (IG) or antibodies, T cell receptors (TR), and immunoglobulin superfamily (IgSF) proteins, and of the groove (G) domains of major histocompatibility (MH; in humans, HLA for human leukocyte antigen) and MH superfamily proteins. IMGT/DomainGapAlign aligns the user own sequences against the IMGT domain reference directory, displays amino acid changes, creates IMGT gaps, and delimits the domain strands and loops (and helix for G domain) according to the IMGT unique numbering. IMGT/DomainGapAlign is coupled to the IMGT/Collier-de-Perles tool that draws standardized IMGT Colliers de Perles. The analysis is based on the IMGT-ONTOLOGY concepts of identification, classification, description, and numerotation generated from the axioms of the Formal IMGT-ONTOLOGY or IMGT-Kaleidoscope. IMGT/DomainGapAlign provides an invaluable help for antibody engineering and antibody humanization as it precisely defines the standardized framework regions (FR-IMGT) and complementarity determining regions (CDR-IMGT) to be grafted. IMGT/DomainGapAlign is freely available at http://www.imgt.org. DOI: 10.1007/978-1-61779-842-9_33 PMID: 22665257 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/308705
1. Surgery. 1978 Oct;84(4):471-5. Purulent pericarditis in children. Garvin PJ, Danis RK, Lewis JE Jr, Willman VL. Acute purulent pericarditis was treated successfully in five children between the ages of 27 months and 11 1/2 years during the past 5 years. The responsible organism was Hemophilus influenzae, type b, in two cases and Meningococcus, Pneumococcus, and coagulase-positive Staphylococcus aureus in one case each. No primary source of infection could be identified in two patients. A high index of suspicion, combined with immediate echocardiograms and pericardiocentesis, led to the diagnosis. Immediate antibiotic therapy was instituted on the basis of the gram stain of the pericardial fluid. All five patients had a pericardial window established--four through subxyphoid approach and the fifth, because of a left pleural effusion, through a left thoracotomy. When the subxyphoid approach was used, sump drains were left for postoperative suction and irrigation. All five patients survived without sequalae during follow-up periods of from 18 months to 5 years. We advocate an aggressive approach to the diagnosis and treatment of this problem. This report documents the safety, ease, and effectiveness of the subxyphoid approach as a means of drainage. PMID: 308705 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24105488
1. J Child Neurol. 2014 Nov;29(11):1562-71. doi: 10.1177/0883073813501870. Epub 2013 Oct 7. Subependymal giant cell astrocytomas in patients with tuberous sclerosis complex: considerations for surgical or pharmacotherapeutic intervention. Wheless JW(1), Klimo P Jr(2). Author information: (1)Department of Pediatric Neurology, Neuroscience Institute and Tuberous Sclerosis Clinic, Le Bonheur Children's Hospital, University of Tennessee Health Science Center, Memphis, TN, USA jwheless@uthsc.edu. (2)Department of Neurosurgery, Neuroscience Institute and Tuberous Sclerosis Clinic, Le Bonheur Children's Hospital, University of Tennessee Health Science Center, Memphis, TN, USA Semmes-Murphey Neurologic & Spine Institute, Memphis, TN, USA St. Jude Children's Research Hospital, Memphis, TN, USA. Tuberous sclerosis complex is a genetic disorder caused by mutations in either the TSC1 or TSC2 gene that can result in the growth of hamartomas in multiple organ systems. Subependymal giant cell astrocytomas are slow-growing brain tumors associated primarily with tuberous sclerosis complex. They are usually located in the ventricles, often near the foramen of Monro, where they can cause an obstruction if they grow too large, leading to increased intracranial pressure. Surgery to remove a tumor has been the mainstay of treatment but can be associated with postoperative morbidity and mortality. Not all tumors and/or patients are suitable for surgery. The recent development of mammalian target of rapamycin inhibitors that target the pathway affected by TSC1/TSC2 mutations offers a novel pharmacotherapeutic option for these patients. We review the timing and use of surgery versus pharmacotherapy for the treatment of subependymal giant cell astrocytoma in patients with tuberous sclerosis complex. © The Author(s) 2013. DOI: 10.1177/0883073813501870 PMID: 24105488 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18503082
1. Nucleic Acids Res. 2008 Jul 1;36(Web Server issue):W503-8. doi: 10.1093/nar/gkn316. Epub 2008 May 24. IMGT/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis. Brochet X(1), Lefranc MP, Giudicelli V. Author information: (1)IMGT, the international ImMunoGeneTics information system, Laboratoire d'ImmunoGénétique Moléculaire LIGM, Université Montpellier 2, Institut de Génétique Humaine IGH, UPR CNRS 1142, 141 rue de la Cardonille, 34396 Montpellier cedex 5, France. IMGT/V-QUEST is the highly customized and integrated system for the standardized analysis of the immunoglobulin (IG) and T cell receptor (TR) rearranged nucleotide sequences. IMGT/V-QUEST identifies the variable (V), diversity (D) and joining (J) genes and alleles by alignment with the germline IG and TR gene and allele sequences of the IMGT reference directory. New functionalities were added through a complete rewrite in Java. IMGT/V-QUEST analyses batches of sequences (up to 50) in a single run. IMGT/V-QUEST describes the V-REGION mutations and identifies the hot spot positions in the closest germline V gene. IMGT/V-QUEST can detect insertions and deletions in the submitted sequences by reference to the IMGT unique numbering. IMGT/V-QUEST integrates IMGT/JunctionAnalysis for a detailed analysis of the V-J and V-D-J junctions, and IMGT/Automat for a full V-J- and V-D-J-REGION annotation. IMGT/V-QUEST displays, in 'Detailed view', the results and alignments for each submitted sequence individually and, in 'Synthesis view', the alignments of the sequences that, in a given run, express the same V gene and allele. The 'Advanced parameters' allow to modify default parameters used by IMGT/V-QUEST and IMGT/JunctionAnalysis according to the users' interest. IMGT/V-QUEST is freely available for academic research at http://imgt.cines.fr. DOI: 10.1093/nar/gkn316 PMCID: PMC2447746 PMID: 18503082 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19900967
1. Nucleic Acids Res. 2010 Jan;38(Database issue):D301-7. doi: 10.1093/nar/gkp946. Epub 2009 Nov 9. IMGT/3Dstructure-DB and IMGT/DomainGapAlign: a database and a tool for immunoglobulins or antibodies, T cell receptors, MHC, IgSF and MhcSF. Ehrenmann F(1), Kaas Q, Lefranc MP. Author information: (1)IMGT, the International imMunoGeneTics Information System, Université Montpellier 2, Laboratoire d'ImmunoGénétique Moléculaire LIGM, Institut de Génétique Humaine IGH, UPR CNRS 1142, 141 rue de la Cardonille, 34396 Montpellier cedex 5, France. IMGT/3Dstructure-DB is the three-dimensional (3D) structure database of IMGT, the international ImMunoGenetics information system that is acknowledged as the global reference in immunogenetics and immunoinformatics. IMGT/3Dstructure-DB contains 3D structures of immunoglobulins (IG) or antibodies, T cell receptors (TR), major histocompatibility complex (MHC) proteins, antigen receptor/antigen complexes (IG/Ag, TR/peptide/MHC) of vertebrates; 3D structures of related proteins of the immune system (RPI) of vertebrates and invertebrates, belonging to the immunoglobulin and MHC superfamilies (IgSF and MhcSF, respectively) and found in complexes with IG, TR or MHC. IMGT/3Dstructure-DB data are annotated according to the IMGT criteria, using IMGT/DomainGapAlign, and based on the IMGT-ONTOLOGY concepts and axioms. IMGT/3Dstructure-DB provides IMGT gene and allele identification (CLASSIFICATION), region and domain delimitations (DESCRIPTION), amino acid positions according to the IMGT unique numbering (NUMEROTATION) that are used in IMGT/3Dstructure-DB cards, results of contact analysis and renumbered flat files. In its Web version, the IMGT/DomainGapAlign tool analyses amino acid sequences, per domain. Coupled to the IMGT/Collier-de-Perles tool, it provides an invaluable help for antibody engineering and humanization design based on complementarity determining region (CDR) grafting as it precisely defines the standardized framework regions (FR-IMGT) and CDR-IMGT. IMGT/3Dstructure-DB and IMGT/DomainGapAlign are freely available at http://www.imgt.org. DOI: 10.1093/nar/gkp946 PMCID: PMC2808948 PMID: 19900967 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17877839
1. BMC Bioinformatics. 2007 Sep 19;8:348. doi: 10.1186/1471-2105-8-348. PADB: published association database. Rhee H(1), Lee JS. Author information: (1)Department of Clinical Genetics, Yonsei University College of Medicine, Seoul, Korea. hwanseok@yonsei.ac.kr BACKGROUND: Although molecular pathway information and the International HapMap Project data can help biomedical researchers to investigate the aetiology of complex diseases more effectively, such information is missing or insufficient in current genetic association databases. In addition, only a few of the environmental risk factors are included as gene-environment interactions, and the risk measures of associations are not indexed in any association databases. DESCRIPTION: We have developed a published association database (PADB; http://www.medclue.com/padb) that includes both the genetic associations and the environmental risk factors available in PubMed database. Each genetic risk factor is linked to a molecular pathway database and the HapMap database through human gene symbols identified in the abstracts. And the risk measures such as odds ratios or hazard ratios are extracted automatically from the abstracts when available. Thus, users can review the association data sorted by the risk measures, and genetic associations can be grouped by human genes or molecular pathways. The search results can also be saved to tab-delimited text files for further sorting or analysis. Currently, PADB indexes more than 1,500,000 PubMed abstracts that include 3442 human genes, 461 molecular pathways and about 190,000 risk measures ranging from 0.00001 to 4878.9. CONCLUSION: PADB is a unique online database of published associations that will serve as a novel and powerful resource for reviewing and interpreting huge association data of complex human diseases. DOI: 10.1186/1471-2105-8-348 PMCID: PMC2039752 PMID: 17877839 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17241641
1. J Mol Biol. 2007 Mar 16;367(1):65-79. doi: 10.1016/j.jmb.2006.10.057. Epub 2006 Oct 21. Phospholamban interacts with HAX-1, a mitochondrial protein with anti-apoptotic function. Vafiadaki E(1), Sanoudou D, Arvanitis DA, Catino DH, Kranias EG, Kontrogianni-Konstantopoulos A. Author information: (1)Molecular Biology Division, Center for Basic Research, Foundation for Biomedical Research of the Academy of Athens, Soranou Efesiou 4, Athens 115 27, Greece. Phospholamban (PLN) is a key regulator of Ca(2+) homeostasis and contractility in the heart. Its regulatory effects are mediated through its interaction with the sarcoplasmic reticulum Ca(2+)-ATPase, (SERCA2a), resulting in alterations of its Ca(2+)-affinity. To identify additional proteins that may interact with PLN, we used the yeast-two-hybrid system to screen an adult human cardiac cDNA library. HS-1 associated protein X-1 (HAX-1) was identified as a PLN-binding partner. The minimal binding regions were mapped to amino acid residues 203-245 for HAX-1 and residues 16-22 for PLN. The interaction between the two proteins was confirmed using GST-HAX-1, bound to the glutathione-matrix, which specifically adsorbed native PLN from human or mouse cardiac homogenates, while in reciprocal binding studies, recombinant His-HAX-1 bound GST-PLN. Kinetic studies using surface plasmon resonance yielded a K(D) of approximately 1 muM as the binding affinity for the PLN/HAX-1 complex. Phosphorylation of PLN by cAMP-dependent protein kinase reduced binding to HAX-1, while increasing concentrations of Ca(2+) diminished the PLN/HAX-1 interaction in a dose-dependent manner. HAX-1 concentrated to mitochondria, but upon transient co-transfection of HEK 293 cells with PLN, HAX-1 redistributed and co-localized with PLN at the endoplasmic reticulum. Analysis of the anti-apoptotic function of HAX-1 revealed that the presence of PLN enhanced the HAX-1 protective effects from hypoxia/reoxygenation-induced cell death. These findings suggest a possible link between the Ca(2+) handling by the sarcoplasmic reticulum and cell survival mediated by the PLN/HAX-1 interaction. DOI: 10.1016/j.jmb.2006.10.057 PMID: 17241641 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/9367782
1. J Mol Biol. 1997 Nov 7;273(4):927-48. doi: 10.1006/jmbi.1997.1354. Standard conformations for the canonical structures of immunoglobulins. Al-Lazikani B(1), Lesk AM, Chothia C. Author information: (1)University of Cambridge Clinical School, Hills Road, Cambridge, CB2 2QH, England. A comparative analysis of the main-chain conformation of the L1, L2, L3, H1 and H2 hypervariable regions in 17 immunoglobulin structures that have been accurately determined at high resolution is described. This involves 79 hypervariable regions in all. We also analysed a part of the H3 region in 12 of the 15 VH domains considered here. On the basis of the residues at key sites the 79 hypervariable regions can be assigned to one of 18 different canonical structures. We show that 71 of these hypervariable regions have a conformation that is very close to what can be defined as a "standard" conformation of each canonical structure. These standard conformations are described in detail. The other eight hypervariable regions have small deviations from the standard conformations that, in six cases, involve only the rotation of a single peptide group. Most H3 hypervariable regions have the same conformation in the part that is close to the framework and the details of this conformation are also described here. Copyright 1997 Academic Press Limited DOI: 10.1006/jmbi.1997.1354 PMID: 9367782 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20505370
1. Plant Signal Behav. 2010 Oct;5(10):1167-70. doi: 10.4161/psb.5.10.11905. Epub 2010 Oct 1. A role for CHROMOMETHYLASE3 in mediating transposon and euchromatin silencing during egg cell reprogramming in Arabidopsis. Pillot M(1), Autran D, Leblanc O, Grimanelli D. Author information: (1)Laboratoire Génome et Développement des Plantes (LGDP), UMR 5096 IRD-CNRS-Université de Perpignan, Montpellier, France. During embryogenesis there is a major switch from dependence upon maternally-deposited products to reliance on products of the zygotic genome. In animals, this so-called maternal-to-zygotic transition occurs following a period of transcriptional quiescence. Recently, we have shown that the early embryo in Arabidopsis is also quiescent, a state inherited from the female gamete and linked to specific patterns of H3K9 dimethylation and TERMINAL FLOWER2 (TFL2) localization. We also demonstrated that CHROMOMETHYLASE 3 (CMT3) is required for H3K9 dimethylation in the egg cell and for normal embryogenesis during the first few divisions of the zygote. Subsequent analysis of CMT3 mutants points to a key role in egg cell reprogramming by controlling silencing in both transposon and euchromatic regions. A speculative model of the CMT3-induced egg cell silencing is presented here, based on these results and current data from the literature suggesting the potential involvement of small RNAs targeted to the egg cell, a process conceptually similar to the division of labor described in the male gametophyte for which we show that H3K9 modifications and TFL2 localization are reminiscent of the female gametophyte. © 2010 Landes Bioscience DOI: 10.4161/psb.5.10.11905 PMCID: PMC3115342 PMID: 20505370 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16820580
1. Circulation. 2006 Jul 4;114(1 Suppl):I245-50. doi: 10.1161/CIRCULATIONAHA.105.000786. Glucose-insulin-potassium and tri-iodothyronine individually improve hemodynamic performance and are associated with reduced troponin I release after on-pump coronary artery bypass grafting. Ranasinghe AM(1), Quinn DW, Pagano D, Edwards N, Faroqui M, Graham TR, Keogh BE, Mascaro J, Riddington DW, Rooney SJ, Townend JN, Wilson IC, Bonser RS. Author information: (1)Department of Cardiothoracic Surgery, University Hospital Birmingham, Edgbaston, Birmingham, B15 2TH, UK. BACKGROUND: Both glucose-insulin-potassium (GIK) and tri-iodothyronine (T3) may improve cardiovascular performance after coronary artery surgery (CABG) but their effects have not been directly compared and the effects of combined treatment are unknown. METHODS AND RESULTS: In 2 consecutive randomized double-blind placebo-controlled trials, in patients undergoing first time isolated on-pump CABG between January 2000 and September 2004, 440 patients were recruited and randomized to either placebo (5% dextrose) (n=160), GIK (40% dextrose, K+ 100 mmol.L(-1), insulin 70 u.L(-1)) (0.75 mL.kg(-1) h(-1)) (n=157), T3 (0.8 microg.kg(-1) followed by 0.113 microg.kg(-1) h(-1)) (n=63) or GIK+T3 (n=60). GIK/placebo therapy was administered from start of operation until 6 hours after removal of aortic cross-clamp (AXC) and T3/placebo was administered for a 6-hour period from removal of AXC. Serial hemodynamic measurements were taken up to 12 hours after removal of AXC and troponin I (cTnI) levels were assayed to 72 hours. Cardiac index (CI) was significantly increased in both the GIK and GIK/T3 group in the first 6 hours compared with placebo (P<0.001 for both) and T3 therapy (P=0.009 and 0.029, respectively). T3 therapy increased CI versus placebo between 6 and 12 hours after AXC removal (P=0.01) but combination therapy did not. Release of cTnI was lower in all treatment groups at 6 and 12 hours after removal of AXC. CONCLUSIONS: Treatment with GIK, T3, and GIK/T3 improves hemodynamic performance and results in reduced cTnI release in patients undergoing on-pump CABG surgery. Combination therapy does not provide added hemodynamic effect. DOI: 10.1161/CIRCULATIONAHA.105.000786 PMID: 16820580 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15946751
1. Biochim Biophys Acta. 2005 Jun 30;1729(2):118-25. doi: 10.1016/j.bbaexp.2005.04.001. Isolation and expression analysis of genes encoding DNA methyltransferase in wheat (Triticum aestivum L.). Dai Y(1), Ni Z, Dai J, Zhao T, Sun Q. Author information: (1)Department of Plant Genetics and Breeding, State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing. DNA methylation of cytosine residues, catalyzed by DNA methyltransferases, is suggested to play important roles in regulating gene expression and plant development. In this study, we isolated four wheat cDNA fragments and one cDNA with open reading frame encoding putative DNA methyltransferase and designated TaMET1, TaMET2a, TaMET2b, TaCMT, TaMET3, respectively. BLASTX searches and phylogenetic analysis suggested that five cDNAs belonged to four classes (Dnmt1, Dnmt2, CMT and Dnmt3) of DNA methyltransferase genes. TaMET2a encoded a protein of 376 aa and contained eight of ten conserved motifs characteristic of DNA methyltransferase. Genomic sequence of TaMET2a was obtained and found to contain ten introns and eleven exons. The expression analysis of the five genes revealed that they were expressed in developing seed, during germination and various vegetative tissues, but in quite different abundance. It was interesting to note that TaMET1 and TaMET3 mRNAs were clearly detected in dry seeds. Moreover, the differential expression patterns of five genes were observed between wheat hybrid and its parents in leaf, stem and root of jointing stage, some were up-regulated while some others were down-regulated in the hybrid. We concluded that multiple wheat DNA methyltransferase genes were present and might play important roles in wheat growth and development. DOI: 10.1016/j.bbaexp.2005.04.001 PMID: 15946751 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21930502
1. Database (Oxford). 2011 Sep 19;2011:bar026. doi: 10.1093/database/bar026. Print 2011. International Cancer Genome Consortium Data Portal--a one-stop shop for cancer genomics data. Zhang J(1), Baran J, Cros A, Guberman JM, Haider S, Hsu J, Liang Y, Rivkin E, Wang J, Whitty B, Wong-Erasmus M, Yao L, Kasprzyk A. Author information: (1)Ontario Institute for Cancer Research, Toronto, Ontario M5G 0A3, Canada. The International Cancer Genome Consortium (ICGC) is a collaborative effort to characterize genomic abnormalities in 50 different cancer types. To make this data available, the ICGC has created the ICGC Data Portal. Powered by the BioMart software, the Data Portal allows each ICGC member institution to manage and maintain its own databases locally, while seamlessly presenting all the data in a single access point for users. The Data Portal currently contains data from 24 cancer projects, including ICGC, The Cancer Genome Atlas (TCGA), Johns Hopkins University, and the Tumor Sequencing Project. It consists of 3478 genomes and 13 cancer types and subtypes. Available open access data types include simple somatic mutations, copy number alterations, structural rearrangements, gene expression, microRNAs, DNA methylation and exon junctions. Additionally, simple germline variations are available as controlled access data. The Data Portal uses a web-based graphical user interface (GUI) to offer researchers multiple ways to quickly and easily search and analyze the available data. The web interface can assist in constructing complicated queries across multiple data sets. Several application programming interfaces are also available for programmatic access. Here we describe the organization, functionality, and capabilities of the ICGC Data Portal. DOI: 10.1093/database/bar026 PMCID: PMC3263593 PMID: 21930502 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20089483
1. Anadolu Kardiyol Derg. 2009 Dec;9 Suppl 2:17-23. Sudden cardiac death in young competitive athletes due to genetic cardiac abnormalities. Wever-Pinzon OE(1), Myerson M, Sherrid MV. Author information: (1)Division of Cardiology, St Luke's - Roosevelt Hospital Center Columbia University, College of Physicians & Surgeons New York City, NY 10019, USA. Sudden cardiac death (SCD) in young athletes is generally caused by inherited cardiac disorders. While these events are relatively few compared to other cardiac deaths, they are tragic in that death occurs in a young, otherwise healthy person. The genetic abnormalities most associated with SCD are hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, long QT syndrome, Brugada syndrome, and catecholaminergic polymorphic ventricular tachycardia. As a result of growing awareness that these deaths can be prevented, guidelines have been issued in both Europe and the United States to help screen and determine qualification for young persons who want to participate in competitive athletics. There remains debate on the how extensive screening should be, in particular over the use of the 12-lead electrocardiogram (ECG), with European guidelines mandating ECG and United States guidelines not recommending routine use of the ECG. PMID: 20089483 [Indexed for MEDLINE]