Cathepsin D is a lysosomal hydrolase involved in intra- and extracellular proteolysis. This enzyme is aberrantly produced and processed in malignancy, and most notably is over-secreted into the tumor cell microenvironment. This hyper-secretion may lead to excessive degradation of the extracellular matrix, and contribute to tumor progression and metastases. These phenomena have been established in vitro, and there is evidence that Cathepsin D is similarly dysregulated in human breast cancer patients. Because breast cancer lacks an effective screening or surveillance biomarker, here we address the hypothesis that serum Cathepsin D activity may be useful to assess the presence or progression of breast cancer in females. While representative histologic sections from various disease-specific cohorts confirm previous findings that increased Cathepsin D production and secretion correlate with tumor progression, we report no difference in serum Cathepsin D activity between patients who are disease free, patients with pre-invasive or limited invasive disease, and patients with metastatic disease. Furthermore, in patients with known metastatic disease, there were no clinical variables associated with significantly different serum Cathepsin D activity. However, the immunohistochemical localization of Cathepsin D expression in histopathologic sections from breast cancer patients correlates with disease progression. Based on the serum results, and in contradistinction to Cathepsin D localization in breast cancer tissues, our findings support using Cathepsin D as a reliable histopathology biomarker for disease progression, but not for serum screening. Abbott, D. E. Margaryan, N. V. Jeruss, J. S. Khan, S. Kaklamani, V. Winchester, D. Hansen, N. Rademaker, A. Khalkhali-Ellis, Z. Hendrix, M. J. 0 2009-11-21T07:19:50Z 2010-01-15 2009-11-20 %0 Journal Article %A Abbott, D. E. %A Margaryan, N. V. %A Jeruss, J. S. %A Khan, S. %A Kaklamani, V. %A Winchester, D. %A Hansen, N. %A Rademaker, A. %A Khalkhali-Ellis, Z. %A Hendrix, M. J. %D 2010 %T Reevaluating cathepsin D as a biomarker for breast cancer: Serum activity levels versus histopathology. %J Cancer Biol Ther %V 9 %N 1 %M 19923884 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19923884 %X Cathepsin D is a lysosomal hydrolase involved in intra- and extracellular proteolysis. This enzyme is aberrantly produced and processed in malignancy, and most notably is over-secreted into the tumor cell microenvironment. This hyper-secretion may lead to excessive degradation of the extracellular matrix, and contribute to tumor progression and metastases. These phenomena have been established in vitro, and there is evidence that Cathepsin D is similarly dysregulated in human breast cancer patients. Because breast cancer lacks an effective screening or surveillance biomarker, here we address the hypothesis that serum Cathepsin D activity may be useful to assess the presence or progression of breast cancer in females. While representative histologic sections from various disease-specific cohorts confirm previous findings that increased Cathepsin D production and secretion correlate with tumor progression, we report no difference in serum Cathepsin D activity between patients who are disease free, patients with pre-invasive or limited invasive disease, and patients with metastatic disease. Furthermore, in patients with known metastatic disease, there were no clinical variables associated with significantly different serum Cathepsin D activity. However, the immunohistochemical localization of Cathepsin D expression in histopathologic sections from breast cancer patients correlates with disease progression. Based on the serum results, and in contradistinction to Cathepsin D localization in breast cancer tissues, our findings support using Cathepsin D as a reliable histopathology biomarker for disease progression, but not for serum screening. %+ Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. 8611 false false 1 Cancer biology & therapy Cancer Biol Ther 2010-01-15 aheadofprint JOURNAL ARTICLE 19923884 Publisher 2009-11-21T07:19:50Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19923884 9 2010 Klippel-Feil syndrome (KFS) is a rare congenital abnormality characterized by a short neck, a low posterior hairline, and limited head movement. Occasionally, patients with KFS may also show signs of deafness, intellectual disability, cardiac malformation, palpebral ptosis, facial nerve paralysis, cleft palate, and scoliosis. Although some researchers have documented this syndrome, scant attention has been paid to craniomaxillofacial manifestations and dental treatment of patients with KFS. The objective of this case report was to describe the planning and execution of dental treatment for a 10-year-old male patient with KFS. de Deus Moura de Lima, M. Ortega, K. L. Araujo, L. C. Soares, M. M. de Magalhaes, M. H. 0 2009-11-07T07:17:49Z 2009-11-06 %0 Journal Article %A de Deus Moura de Lima, M. %A Ortega, K. L. %A Araujo, L. C. %A Soares, M. M. %A de Magalhaes, M. H. %D 2009 %T Dental team management for a patient with Klippel-Feil syndrome: Case report. %J Spec Care Dentist %V 29 %N 6 %P 244-248 %M 19886936 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19886936 %X Klippel-Feil syndrome (KFS) is a rare congenital abnormality characterized by a short neck, a low posterior hairline, and limited head movement. Occasionally, patients with KFS may also show signs of deafness, intellectual disability, cardiac malformation, palpebral ptosis, facial nerve paralysis, cleft palate, and scoliosis. Although some researchers have documented this syndrome, scant attention has been paid to craniomaxillofacial manifestations and dental treatment of patients with KFS. The objective of this case report was to describe the planning and execution of dental treatment for a 10-year-old male patient with KFS. %+ Postgraduate Student, Special Care Dentistry Center, Department of Oral Pathology, University of Sao Paulo, Sao Paulo, Brazil. de Deus Moura de Lima, Marina Ortega, Karem Lopez Araujo, Luis Carlos Arias Soares, Marcelo Melo de Magalhaes, Marina Helena Cury Gallottini 8570 false false 6 Special care in dentistry : official publication of the American Association of Hospital Dentists, the Academy of Dentistry for the Handicapped, and the American Society for Geriatric Dentistry Spec Care Dentist 244-248 ppublish Journal Article 19886936 In-Data-Review 2009-11-07T07:17:49Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19886936 29 2009 Iannaccone, P. M. Jacob, H. J. 0 2009-09-06T11:55:40Z 2009-05-02 %0 Journal Article %A Iannaccone, P. M. %A Jacob, H. J. %D 2009 %T Rats! %J Dis Model Mech %V 2 %N 5-6 %P 206-210 %M 19407324 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19407324 %K Animals %K *Disease Models, Animal %K Embryonic Stem Cells/cytology %K Humans %K Mice %K Rats Iannaccone, Philip M Jacob, Howard J 2412 false false 5-6 Disease models & mechanisms Dis Model Mech Animals; *Disease Models, Animal; Embryonic Stem Cells/cytology; Humans; Mice; Rats 206-210 ppublish Editorial 19407324 MEDLINE 2009-09-06T11:55:40Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19407324 2 2009 PURPOSE OF REVIEW: Food allergy, a growing clinical and public health problem in the United States and worldwide, is likely determined by multiple environmental and genetic factors. The purpose of this review is to summarize recent advances in food allergy genetic research. RECENT FINDINGS: There is compelling evidence that genetic factors may play a role in food allergy. However, the specific genetic loci that may modulate individual risk of food allergy remain to be identified. To date, only a limited number of candidate gene association studies of food allergy have been reported. Polymorphism(s) in nine genes have been associated with the incidence of food allergy or food allergy severity in at least one study. But most of these findings remain to be replicated in independent populations. In contrast, there are considerable advances in genetics of other allergic diseases such as asthma and atopic dermatitis. Although asthma and atopic dermatitis often coexist with food allergy, the relevance of their candidate genes to food allergy remains to be evaluated. SUMMARY: Genetics in food allergy is a promising research area but is still in its infancy. More studies are needed to dissect susceptible genes of food allergy. A genome-wide association approach may serve as a powerful tool to identify novel genes related to food allergy. Furthermore, the role of gene-environment interaction, gene-gene interaction, and epigenetics in food allergy remains largely unexplored. Given the complex nature of food allergy, future studies need to integrate environment, genomics, and epigenomics in order to better understand the multifaceted etiology and biological mechanisms of food allergy. Hong, X. Tsai, H. J. Wang, X. 0 2009-10-25T06:20:54Z 2009-10-24 %0 Journal Article %A Hong, X. %A Tsai, H. J. %A Wang, X. %D 2009 %T Genetics of food allergy. %J Curr Opin Pediatr %V 21 %N 6 %P 770-776 %M 19851108 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19851108 %X PURPOSE OF REVIEW: Food allergy, a growing clinical and public health problem in the United States and worldwide, is likely determined by multiple environmental and genetic factors. The purpose of this review is to summarize recent advances in food allergy genetic research. RECENT FINDINGS: There is compelling evidence that genetic factors may play a role in food allergy. However, the specific genetic loci that may modulate individual risk of food allergy remain to be identified. To date, only a limited number of candidate gene association studies of food allergy have been reported. Polymorphism(s) in nine genes have been associated with the incidence of food allergy or food allergy severity in at least one study. But most of these findings remain to be replicated in independent populations. In contrast, there are considerable advances in genetics of other allergic diseases such as asthma and atopic dermatitis. Although asthma and atopic dermatitis often coexist with food allergy, the relevance of their candidate genes to food allergy remains to be evaluated. SUMMARY: Genetics in food allergy is a promising research area but is still in its infancy. More studies are needed to dissect susceptible genes of food allergy. A genome-wide association approach may serve as a powerful tool to identify novel genes related to food allergy. Furthermore, the role of gene-environment interaction, gene-gene interaction, and epigenetics in food allergy remains largely unexplored. Given the complex nature of food allergy, future studies need to integrate environment, genomics, and epigenomics in order to better understand the multifaceted etiology and biological mechanisms of food allergy. %+ Mary Ann and J. Milburn Smith Child Health Research Program, Department of Pediatrics, Northwestern University Feinberg School of Medicine and Children's Memorial Hospital and Children's Memorial Research Center, Chicago, Illinois, USA. 8526 false false 6 Current opinion in pediatrics Curr Opin Pediatr 770-776 2009-12-01 ppublish JOURNAL ARTICLE 19851108 Publisher 2009-11-19T07:23:14Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19851108 21 2009 In the 1970s, several human retinoblastoma cell lines were developed from cultures of primary tumors. As the human retinoblastoma cell lines were established in culture, growth properties and changes in cell adhesion were described. Those changes correlated with the ability of the human retinoblastoma cell lines to invade the optic nerve and metastasize in orthotopic xenograft studies. However, the mechanisms that underly these changes were not determined. We have used the recently developed knockout mouse models of retinoblastoma to begin to characterize the molecular, cellular and genetic changes associated with retinoblastoma tumor progression and optic nerve invasion. Here we present the isolation and characterization of the first mouse retinoblastoma cell lines with targeted deletions of the Rb family. Our detailed analysis of these cells as they were propagated in culture from the primary tumor show that changes in cadherin-mediated cell adhesion are associated with retinoblastoma invasion of the optic nerve prior to metastasis. In addition, the same changes in cadherin-mediated cell adhesion correlate with the invasive properties of the human retinoblastoma cell lines isolated decades ago providing a molecular mechanism for these earlier observations. Most importantly, our studies are in agreement with genetic studies on human retinoblastomas suggesting that changes in this pathway are involved in tumor progression. Laurie, N. Mohan, A. McEvoy, J. Reed, D. Zhang, J. Schweers, B. Ajioka, I. Valentine, V. Johnson, D. Ellison, D. Dyer, M. A. 0 2009-10-01T06:13:58Z 2009-09-28 2009-09-30 %0 Journal Article %A Laurie, N. %A Mohan, A. %A McEvoy, J. %A Reed, D. %A Zhang, J. %A Schweers, B. %A Ajioka, I. %A Valentine, V. %A Johnson, D. %A Ellison, D. %A Dyer, M. A. %D 2009 %T Changes in retinoblastoma cell adhesion associated with optic nerve invasion. %J Mol Cell Biol %V 29 %N 23 %P 6268-6282 %M 19786571 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19786571 %X In the 1970s, several human retinoblastoma cell lines were developed from cultures of primary tumors. As the human retinoblastoma cell lines were established in culture, growth properties and changes in cell adhesion were described. Those changes correlated with the ability of the human retinoblastoma cell lines to invade the optic nerve and metastasize in orthotopic xenograft studies. However, the mechanisms that underlie these changes were not determined. We used the recently developed knockout mouse models of retinoblastoma to begin to characterize the molecular, cellular, and genetic changes associated with retinoblastoma tumor progression and optic nerve invasion. Here we report the isolation and characterization of the first mouse retinoblastoma cell lines with targeted deletions of the Rb family. Our detailed analysis of these cells as they were propagated in culture from the primary tumor shows that changes in cadherin-mediated cell adhesion are associated with retinoblastoma invasion of the optic nerve prior to metastasis. In addition, the same changes in cadherin-mediated cell adhesion correlate with the invasive properties of the human retinoblastoma cell lines isolated decades ago, providing a molecular mechanism for these earlier observations. Most importantly, our studies are in agreement with genetic studies on human retinoblastomas, suggesting that changes in this pathway are involved in tumor progression. %+ Department of Developmental Neurobiology, MS 323, St. Jude Children's Research Hospital, Memphis, TN 38105, USA. 8422 false false 23 Molecular and cellular biology Mol Cell Biol 6268-6282 2009-12-01 ppublish JOURNAL ARTICLE 19786571 In-Process 2009-11-10T07:18:39Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19786571 29 2009 Tyrosine kinase inhibitors, such as imatinib, have dramatically improved the outcomes for patients with selected cancers. For imatinib, western blotting of phospho-CrkL was an insensitive, indirect, and descriptive method to determine drug efficacy. Greater use of targeted therapies should involve more quantitative evaluation of the target's dose-inhibition. The Src/Abl kinase inhibitor dasatinib has recently been approved for use in Ph+ leukemias after failure with imatinib. Src family kinases (SFK) also play a critical role in nonhematologic cancers. We have developed a flow cytometric assay to measure SFK autophosphorylation levels in blood mononuclear cells and observed a direct correlation between its inhibition and patient dosage. This method provides a sensitive, quick, and quantitative tool to assess drug efficacy. Pediatr Blood Cancer. (c) 2009 Wiley-Liss, Inc. Guerrouahen, B. S. Wieder, E. Blanchard, E. G. Lee, F. Y. Aplenc, R. Corey, S. J. 0 2009-09-06T12:00:16Z 2009-06-02 %0 Journal Article %A Guerrouahen, B. S. %A Wieder, E. %A Blanchard, E. G. %A Lee, F. Y. %A Aplenc, R. %A Corey, S. J. %D 2009 %T Flow cytometric determination of Src phosphorylation in pediatric patients treated with dasatinib. %J Pediatr Blood Cancer %V 53 %N 6 %P 1132-1135 %M 19484755 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19484755 %X Tyrosine kinase inhibitors, such as imatinib, have dramatically improved the outcomes for patients with selected cancers. For imatinib, western blotting of phospho-CrkL was an insensitive, indirect, and descriptive method to determine drug efficacy. Greater use of targeted therapies should involve more quantitative evaluation of the target's dose-inhibition. The Src/Abl kinase inhibitor dasatinib has recently been approved for use in Ph+ leukemias after failure with imatinib. Src family kinases (SFK) also play a critical role in nonhematologic cancers. We have developed a flow cytometric assay to measure SFK autophosphorylation levels in blood mononuclear cells and observed a direct correlation between its inhibition and patient dosage. This method provides a sensitive, quick, and quantitative tool to assess drug efficacy. %K Child %K Flow Cytometry %K Humans %K Methods %K Phosphorylation %K Pyrimidines/*therapeutic use %K Thiazoles/*therapeutic use %K src-Family Kinases/*metabolism %+ Division of Pediatrics, MD Anderson Cancer Center, Houston, Texas, USA. 7399 false false 6 Pediatric blood & cancer Pediatr Blood Cancer Child; Flow Cytometry; Humans; Methods; Phosphorylation; Pyrimidines/*therapeutic use; Thiazoles/*therapeutic use; src-Family Kinases/*metabolism 1132-1135 2009-12-01 ppublish JOURNAL ARTICLE 19484755 MEDLINE 2009-10-14T06:15:31Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19484755 53 2009 The newly described F-BAR (Fer/CIP4 and Bin, amphiphysin, Rvs) family of proteins includes Cdc42 interacting protein-4 (CIP4), FBP-17 and Toca-1, and drives membrane deformation and invagination. Membrane remodeling affects endocytosis, vesicle budding, and cargo selection. The F-BAR family presents a novel family of proteins, which little is known about their in vivo function. We investigated the physiological role of CIP4, by creating CIP4-null mice through homologous recombination. Compared to their wild-type littermates, the CIP4-null mice displayed lower early post-prandial glucose levels. Adipocytes isolated from CIP4-null mice exhibited increased 14C-2-deoxyglucose uptake compared to cells from wild-type mice. The enhanced insulin sensitivity was not due to higher levels of insulin or phospho-Akt, a critical player in the insulin signaling. However, higher GLUT4 levels were detected in muscle membrane fractions in CIP4-null mice under insulin stimulation. Mouse embryonic fibroblasts from CIP4-null mice demonstrated decreased transferrin uptake, FITC-dextran, and horseradish peroxidase uptake, indicating that CIP4 affects multiple modes of endocytosis. These studies demonstrate a physiological role for CIP4 in endocytosis leading to a whole animal phenotype. Feng, Y. Hartig, S. M. Bechill, J. E. Blanchard, E. G. Caudell, E. Corey, S. J. 0 2009-11-20T07:19:25Z 2009-11-17 2009-11-19 %0 Journal Article %A Feng, Y. %A Hartig, S. M. %A Bechill, J. E. %A Blanchard, E. G. %A Caudell, E. %A Corey, S. J. %D 2009 %T The Cdc42 interacting protein 4 (CIP4) gene knockout mouse reveals delayed and decreased endocytosis. %J J Biol Chem %M 19920150 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19920150 %X The newly described F-BAR (Fer/CIP4 and Bin, amphiphysin, Rvs) family of proteins includes Cdc42 interacting protein-4 (CIP4), FBP-17 and Toca-1, and drives membrane deformation and invagination. Membrane remodeling affects endocytosis, vesicle budding, and cargo selection. The F-BAR family presents a novel family of proteins, which little is known about their in vivo function. We investigated the physiological role of CIP4, by creating CIP4-null mice through homologous recombination. Compared to their wild-type littermates, the CIP4-null mice displayed lower early post-prandial glucose levels. Adipocytes isolated from CIP4-null mice exhibited increased 14C-2-deoxyglucose uptake compared to cells from wild-type mice. The enhanced insulin sensitivity was not due to higher levels of insulin or phospho-Akt, a critical player in the insulin signaling. However, higher GLUT4 levels were detected in muscle membrane fractions in CIP4-null mice under insulin stimulation. Mouse embryonic fibroblasts from CIP4-null mice demonstrated decreased transferrin uptake, FITC-dextran, and horseradish peroxidase uptake, indicating that CIP4 affects multiple modes of endocytosis. These studies demonstrate a physiological role for CIP4 in endocytosis leading to a whole animal phenotype. %+ Northwestern, United States; 8610 false false The Journal of biological chemistry J Biol Chem 2009-11-17 aheadofprint JOURNAL ARTICLE 19920150 Publisher 2009-11-20T07:19:25Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19920150 2009 We have recently discovered that de-N-acetyl GM3 [NeuNH(2)LacCer, d-GM3], a derivative of ganglioside GM3, is specifically expressed in metastatic tumor cells and that its expression correlates with an enhanced metastatic phenotype. Although the classic N-acetylated form of GM3 (NeuAcLacCer, c-GM3) is found in both normal and tumor cells, metastatic tumor cells (but not other cells) predominantly express d-GM3 (82-95% of total GM3). d-GM3 expression is mainly found in metastatic melanomas, but not in benign nevi or the majority of primary melanomas. Using metastatic (d-GM3-positive) and poorly invasive (d-GM3-negative) human melanoma cell lines, we found that d-GM3 stimulates cell migration and invasion by increasing the expression and activation of urokinase-like plasminogen activator (uPA). Further studies showed that d-GM3 activates matrix metalloproteinase-2 (MMP-2), but not MMP-9, when uPA receptor signaling is activated. These results implicate d-GM3 as a specific marker for metastatic melanoma and a novel therapeutic target for neoplastic diseases. [Cancer Res 2009;69(22):8662-9]. Liu, J. W. Sun, P. Yan, Q. Paller, A. S. Gerami, P. Ho, N. Vashi, N. Le Poole, I. C. Wang, X. Q. 0 2009-11-13T07:19:52Z 2009-11-10 2009-11-12 %0 Journal Article %A Liu, J. W. %A Sun, P. %A Yan, Q. %A Paller, A. S. %A Gerami, P. %A Ho, N. %A Vashi, N. %A Le Poole, I. C. %A Wang, X. Q. %D 2009 %T De-N-acetyl GM3 promotes melanoma cell migration and invasion through urokinase plasminogen activator receptor signaling-dependent MMP-2 activation. %J Cancer Res %V 69 %N 22 %P 8662-8669 %M 19903858 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19903858 %X We have recently discovered that de-N-acetyl GM3 [NeuNH(2)LacCer, d-GM3], a derivative of ganglioside GM3, is specifically expressed in metastatic tumor cells and that its expression correlates with an enhanced metastatic phenotype. Although the classic N-acetylated form of GM3 (NeuAcLacCer, c-GM3) is found in both normal and tumor cells, metastatic tumor cells (but not other cells) predominantly express d-GM3 (82-95% of total GM3). d-GM3 expression is mainly found in metastatic melanomas, but not in benign nevi or the majority of primary melanomas. Using metastatic (d-GM3-positive) and poorly invasive (d-GM3-negative) human melanoma cell lines, we found that d-GM3 stimulates cell migration and invasion by increasing the expression and activation of urokinase-like plasminogen activator (uPA). Further studies showed that d-GM3 activates matrix metalloproteinase-2 (MMP-2), but not MMP-9, when uPA receptor signaling is activated. These results implicate d-GM3 as a specific marker for metastatic melanoma and a novel therapeutic target for neoplastic diseases. %+ Department of Oncology, the First Affiliated Hospital, Dalian Medical University, Dalian, Liaoning, China. 8588 false false 22 Cancer research Cancer Res 8662-8669 2009-11-15 ppublish JOURNAL ARTICLE 19903858 In-Process 2009-11-17T07:21:11Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19903858 69 2009 PURPOSE: Recent studies suggest that children <24 months with stage I favorable histology Wilms tumors <550 g [very low risk Wilms tumors (VLRWT)] have an excellent prognosis when treated with nephrectomy only, without adjuvant chemotherapy. The identification of risk categories within VLRWT may enable refinement of their definition and optimization of their therapy. EXPERIMENTAL DESIGN: To define biologically distinct subsets, global gene expression analysis was done on 39 VLRWT that passed all quality-control parameters and the clusters identified were validated in an independent set of 11 VLRWT. Validation of select differentially expressed genes was done with immunohistochemistry on a tissue microarray from 20 of 39 tumors. Loss of heterozygosity (LOH) for 11p15, 1p, and 16q was analyzed in 52 tumors using PCR. RESULTS: Two distinctive clusters were identified. One cluster included 9 tumors with epithelial differentiated tubular histology, paucity of nephrogenic rests, lack of LOH for 1p, 16q, and 11p, absence of relapse, and a unique gene expression profile consistent with arrest following mesenchymal-to-epithelial transition. The second cluster included 13 tumors with mixed histology, intralobar nephrogenic rests, and decreased expression of WT1. Three of 6 relapses occurred in this cluster. Of 43 informative tumors, 11p LOH was present in 5 of 5 relapses and 11 of 38 nonrelapses. CONCLUSIONS: Two subsets comprising a total of 56% of VLRWT are identified that have pathogenetic and molecular differences and apparent differences in risk for relapse. If these predictors can be prospectively validated, this would enable the refinement of clinical stratification and less arbitrary definition of VLRWT. (Clin Cancer Res 2009;15(22):6800-9). Sredni, S. T. Gadd, S. Huang, C. C. Breslow, N. Grundy, P. Green, D. M. Dome, J. S. Shamberger, R. C. Beckwith, J. B. Perlman, E. J. 0 2009-11-13T07:19:53Z 2009-11-10 2009-11-12 %0 Journal Article %A Sredni, S. T. %A Gadd, S. %A Huang, C. C. %A Breslow, N. %A Grundy, P. %A Green, D. M. %A Dome, J. S. %A Shamberger, R. C. %A Beckwith, J. B. %A Perlman, E. J. %D 2009 %T Subsets of very low risk wilms tumor show distinctive gene expression, histologic, and clinical features. %J Clin Cancer Res %V 15 %N 22 %P 6800-6809 %M 19903788 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19903788 %X PURPOSE: Recent studies suggest that children <24 months with stage I favorable histology Wilms tumors <550 g [very low risk Wilms tumors (VLRWT)] have an excellent prognosis when treated with nephrectomy only, without adjuvant chemotherapy. The identification of risk categories within VLRWT may enable refinement of their definition and optimization of their therapy. EXPERIMENTAL DESIGN: To define biologically distinct subsets, global gene expression analysis was done on 39 VLRWT that passed all quality-control parameters and the clusters identified were validated in an independent set of 11 VLRWT. Validation of select differentially expressed genes was done with immunohistochemistry on a tissue microarray from 20 of 39 tumors. Loss of heterozygosity (LOH) for 11p15, 1p, and 16q was analyzed in 52 tumors using PCR. RESULTS: Two distinctive clusters were identified. One cluster included 9 tumors with epithelial differentiated tubular histology, paucity of nephrogenic rests, lack of LOH for 1p, 16q, and 11p, absence of relapse, and a unique gene expression profile consistent with arrest following mesenchymal-to-epithelial transition. The second cluster included 13 tumors with mixed histology, intralobar nephrogenic rests, and decreased expression of WT1. Three of 6 relapses occurred in this cluster. Of 43 informative tumors, 11p LOH was present in 5 of 5 relapses and 11 of 38 nonrelapses. CONCLUSIONS: Two subsets comprising a total of 56% of VLRWT are identified that have pathogenetic and molecular differences and apparent differences in risk for relapse. If these predictors can be prospectively validated, this would enable the refinement of clinical stratification and less arbitrary definition of VLRWT. %+ Departments of Pathology and Preventive Medicine, Northwestern University Feinberg School of Medicine and Robert H. Lurie Cancer Center, Chicago, Illinois, USA. 8589 false false 22 Clinical cancer research : an official journal of the American Association for Cancer Research Clin Cancer Res 6800-6809 2009-11-15 ppublish JOURNAL ARTICLE 19903788 In-Process 2009-11-17T07:21:12Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19903788 15 2009 BACKGROUND: Trilateral retinoblastoma has been lethal in virtually all cases previously reported. We describe a series of 13 patients treated with intensive chemotherapy, defined as the intention to include high-dose chemotherapy with autologous hematopoietic stem cell rescue. PROCEDURE: Induction chemotherapy generally included vincristine, cisplatin or carboplatin, cyclophosphamide, and etoposide. Hematopoietic stem cells typically were harvested after the first or second cycle of induction chemotherapy, usually from peripheral blood. High-dose chemotherapy regimens were thiotepa-based (n = 7) or melphalan and cyclophosphamide (n = 3). RESULTS: Trilateral sites were pineal (n = 11) and suprasellar (n = 2); 7 patients had localized (M-0) disease and six had leptomeningeal dissemination (M-1+). Five patients had trilateral retinoblastoma at original diagnosis of intra-ocular retinoblastoma; eight later developed trilateral disease at a median of 35 months (range 3-60 months) following diagnosis of intra-ocular retinoblastoma. One patient died of toxicity (septicemia and multi-organ system failure) during induction and three developed disease progression prior to high-dose chemotherapy. Nine patients received high-dose chemotherapy at a median of 5 months (range 4-9) post-diagnosis of trilateral disease. Five patients survive event-free at a median of 77 months (range 36-104 months) and never received external beam radiation therapy. Four of seven patients with M-0 disease survive event-free versus only one of six patients with M-1+ disease. CONCLUSIONS: Intensive chemotherapy is potentially curative for some patients with trilateral retinoblastoma, especially those with M-0 disease Pediatr Blood Cancer. (c) 2009 Wiley-Liss, Inc. Dunkel, I. J. Jubran, R. F. Gururangan, S. Chantada, G. L. Finlay, J. L. Goldman, S. Khakoo, Y. O'Brien, J. M. Orjuela, M. Rodriguez-Galindo, C. Souweidane, M. M. Abramson, D. H. 0 2009-11-14T07:16:23Z 2009-11-11 2009-11-13 %0 Journal Article %A Dunkel, I. J. %A Jubran, R. F. %A Gururangan, S. %A Chantada, G. L. %A Finlay, J. L. %A Goldman, S. %A Khakoo, Y. %A O'Brien, J. M. %A Orjuela, M. %A Rodriguez-Galindo, C. %A Souweidane, M. M. %A Abramson, D. H. %D 2009 %T Trilateral retinoblastoma: Potentially curable with intensive chemotherapy. %J Pediatr Blood Cancer %M 19908299 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19908299 %X BACKGROUND: Trilateral retinoblastoma has been lethal in virtually all cases previously reported. We describe a series of 13 patients treated with intensive chemotherapy, defined as the intention to include high-dose chemotherapy with autologous hematopoietic stem cell rescue. PROCEDURE: Induction chemotherapy generally included vincristine, cisplatin or carboplatin, cyclophosphamide, and etoposide. Hematopoietic stem cells typically were harvested after the first or second cycle of induction chemotherapy, usually from peripheral blood. High-dose chemotherapy regimens were thiotepa-based (n = 7) or melphalan and cyclophosphamide (n = 3). RESULTS: Trilateral sites were pineal (n = 11) and suprasellar (n = 2); 7 patients had localized (M-0) disease and six had leptomeningeal dissemination (M-1+). Five patients had trilateral retinoblastoma at original diagnosis of intra-ocular retinoblastoma; eight later developed trilateral disease at a median of 35 months (range 3-60 months) following diagnosis of intra-ocular retinoblastoma. One patient died of toxicity (septicemia and multi-organ system failure) during induction and three developed disease progression prior to high-dose chemotherapy. Nine patients received high-dose chemotherapy at a median of 5 months (range 4-9) post-diagnosis of trilateral disease. Five patients survive event-free at a median of 77 months (range 36-104 months) and never received external beam radiation therapy. Four of seven patients with M-0 disease survive event-free versus only one of six patients with M-1+ disease. CONCLUSIONS: Intensive chemotherapy is potentially curative for some patients with trilateral retinoblastoma, especially those with M-0 disease Pediatr Blood Cancer. (c) 2009 Wiley-Liss, Inc. %+ Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, New York; 8590 false false Pediatric blood & cancer Pediatr Blood Cancer 2009-11-11 aheadofprint JOURNAL ARTICLE 19908299 Publisher 2009-11-14T07:16:23Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19908299 2009 We report on a child with chronic granulomatous disease who at the age of 13 years was diagnosed with glioblastoma multiforme of the left thalamus. Therapy included subtotal resection, radiation to the tumor bed (60 Gy), and concomitant chemotherapy with daily thalidomide (250 mg/m), both during radiation and for 5 years thereafter. Currently, she is 9 years from diagnosis and has no evidence of disease. Therapy with thalidomide did not increase her infection complications and provided excellent quality of life. This is the first report of glioblastoma multiforme in a patient with chronic granulomatous disease treated with surgery, radiation, and thalidomide who is a long-term survivor. Aguilera, D. G. Tomita, T. Rajaram, V. Fangusaro, J. Katz, B. Z. Shulman, S. Goldman, S. 0 2009-11-07T07:17:08Z 2009-11-11 2009-11-06 %0 Journal Article %A Aguilera, D. G. %A Tomita, T. %A Rajaram, V. %A Fangusaro, J. %A Katz, B. Z. %A Shulman, S. %A Goldman, S. %D 2009 %T Glioblastoma Multiforme in a Patient With Chronic Granulomatous Disease Treated With Subtotal Resection, Radiation, and Thalidomide: Case Report of a Long-term Survivor. %J J Pediatr Hematol Oncol %M 19887959 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19887959 %X We report on a child with chronic granulomatous disease who at the age of 13 years was diagnosed with glioblastoma multiforme of the left thalamus. Therapy included subtotal resection, radiation to the tumor bed (60 Gy), and concomitant chemotherapy with daily thalidomide (250 mg/m), both during radiation and for 5 years thereafter. Currently, she is 9 years from diagnosis and has no evidence of disease. Therapy with thalidomide did not increase her infection complications and provided excellent quality of life. This is the first report of glioblastoma multiforme in a patient with chronic granulomatous disease treated with surgery, radiation, and thalidomide who is a long-term survivor. %+ Divisions of *Hematology-Oncology and Stem Cell transplantation daggerNeurosurgery section signInfectious Diseases double daggerDepartment of Pathology, Children's Memorial Hospital, Feinberg School of Medicine, Northwestern University, Chicago, IL. 8569 false false Journal of pediatric hematology/oncology : official journal of the American Society of Pediatric Hematology/Oncology J Pediatr Hematol Oncol 2009-11-11 aheadofprint JOURNAL ARTICLE 19887959 Publisher 2009-11-17T07:20:47Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19887959 2009 BACKGROUND: Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder characterized by a decreased ability to repair DNA damaged by UV radiation and the early development of cutaneous and ocular malignant neoplasms. Approximately 20% of patients with XP also develop progressive neurologic degeneration. OBSERVATIONS: We describe a boy who was found to have XP after a severe burn following minimal sun exposure. His maternal uncle, now age 20 years, had been diagnosed with XP after a similar sunburn in infancy. The uncle has the typical skin pigmentary findings of XP along with severe progressive neurologic involvement. Although the infant's parents were not known to be blood relatives, the infant and his affected uncle proved to be compound heterozygotes for the same 2 frameshift mutations in the XPA DNA repair gene (c.288delT and c.349_353del). After the diagnosis of XP in the infant, genealogic investigation identified a common Dutch ancestor for both of his grandfathers 5 generations back. CONCLUSIONS: Counseling families at risk for a rare inherited disease is not always straightforward. The sociocultural and demographic backgrounds of the families must be considered for evaluation of risk assessment. Christen-Zaech, S. Imoto, K. Khan, S. G. Oh, K. S. Tamura, D. Digiovanna, J. J. Boyle, J. Patronas, N. J. Schiffmann, R. Kraemer, K. H. Paller, A. S. 0 2009-11-19T07:22:46Z 2009-11-18 %0 Journal Article %A Christen-Zaech, S. %A Imoto, K. %A Khan, S. G. %A Oh, K. S. %A Tamura, D. %A Digiovanna, J. J. %A Boyle, J. %A Patronas, N. J. %A Schiffmann, R. %A Kraemer, K. H. %A Paller, A. S. %D 2009 %T Unexpected occurrence of xeroderma pigmentosum in an uncle and nephew. %J Arch Dermatol %V 145 %N 11 %P 1285-1291 %M 19917958 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19917958 %X BACKGROUND: Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder characterized by a decreased ability to repair DNA damaged by UV radiation and the early development of cutaneous and ocular malignant neoplasms. Approximately 20% of patients with XP also develop progressive neurologic degeneration. OBSERVATIONS: We describe a boy who was found to have XP after a severe burn following minimal sun exposure. His maternal uncle, now age 20 years, had been diagnosed with XP after a similar sunburn in infancy. The uncle has the typical skin pigmentary findings of XP along with severe progressive neurologic involvement. Although the infant's parents were not known to be blood relatives, the infant and his affected uncle proved to be compound heterozygotes for the same 2 frameshift mutations in the XPA DNA repair gene (c.288delT and c.349_353del). After the diagnosis of XP in the infant, genealogic investigation identified a common Dutch ancestor for both of his grandfathers 5 generations back. CONCLUSIONS: Counseling families at risk for a rare inherited disease is not always straightforward. The sociocultural and demographic backgrounds of the families must be considered for evaluation of risk assessment. %+ Department of Dermatology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Christen-Zaech, Stephanie Imoto, Kyoko Khan, Sikandar G Oh, Kyu-Seon Tamura, Deborah Digiovanna, John J Boyle, Jennifer Patronas, Nickolas J Schiffmann, Raphael Kraemer, Kenneth H Paller, Amy S 8607 false false 11 Archives of dermatology Arch Dermatol 1285-1291 2009-11-01 ppublish Journal Article 19917958 In-Process 2009-11-19T07:22:46Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19917958 145 2009 Therapy-related myelodysplastic syndrome/acute myeloid leukemia (t-MDS/AML) is a long-term complication of pediatric cancer. We retrospectively studied pediatric t-MDS/AML patients treated at MD Anderson from 1975 to 2007. We also compared those patients to pediatric patients with de novo MDS/AML during this time interval. Among 2589 children with cancer treated at MD Anderson, we identified 22 patients with t-MDS/AML. Patients with t-MDS/AML had a median age of 14 years. There was a male and Hispanic predominance. The most common primary malignancies were osteosarcoma and Hodgkin lymphoma. The median latency period was 4.1 years. Three patients received supportive care only. Group 1 (n=5) underwent stem cell transplantation without induction chemotherapy. Group 2 (n=5) patients received AML-type chemotherapy and a stem cell transplant postremission (n=5). Group 3 (n=4) received a stem cell transplant as salvage therapy. The respective 2-year survival rates for groups 1, 2, and 3 were 20%, 40%, and 25% (P=0.85). Patients with de novo AML were younger (P=0.001) and higher rates of complete remission (P=0.03), and survival (P<0.0001). Independent factors predicting shorter survival were poor/intermediate-risk cytogenetics (P=0.01), lower hemoglobin level (P=0.0001), and t-MDS/AML (vs. de novo) (P=0.003). Childhood t-MDS/AML has a poor prognosis. Although patients benefited from AML-type induction chemotherapy followed by stem cell transplantation as postremission therapy, effective therapies, and prevention are needed. Aguilera, D. G. Vaklavas, C. Tsimberidou, A. M. Wen, S. Medeiros, L. J. Corey, S. J. 0 2009-10-07T06:13:28Z 2009-10-06 %0 Journal Article %A Aguilera, D. G. %A Vaklavas, C. %A Tsimberidou, A. M. %A Wen, S. %A Medeiros, L. J. %A Corey, S. J. %D 2009 %T Pediatric therapy-related myelodysplastic syndrome/acute myeloid leukemia: the MD Anderson Cancer Center experience. %J J Pediatr Hematol Oncol %V 31 %N 11 %P 803-811 %M 19801947 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19801947 %X Therapy-related myelodysplastic syndrome/acute myeloid leukemia (t-MDS/AML) is a long-term complication of pediatric cancer. We retrospectively studied pediatric t-MDS/AML patients treated at MD Anderson from 1975 to 2007. We also compared those patients to pediatric patients with de novo MDS/AML during this time interval. Among 2589 children with cancer treated at MD Anderson, we identified 22 patients with t-MDS/AML. Patients with t-MDS/AML had a median age of 14 years. There was a male and Hispanic predominance. The most common primary malignancies were osteosarcoma and Hodgkin lymphoma. The median latency period was 4.1 years. Three patients received supportive care only. Group 1 (n=5) underwent stem cell transplantation without induction chemotherapy. Group 2 (n=5) patients received AML-type chemotherapy and a stem cell transplant postremission (n=5). Group 3 (n=4) received a stem cell transplant as salvage therapy. The respective 2-year survival rates for groups 1, 2, and 3 were 20%, 40%, and 25% (P=0.85). Patients with de novo AML were younger (P=0.001) and higher rates of complete remission (P=0.03), and survival (P<0.0001). Independent factors predicting shorter survival were poor/intermediate-risk cytogenetics (P=0.01), lower hemoglobin level (P=0.0001), and t-MDS/AML (vs. de novo) (P=0.003). Childhood t-MDS/AML has a poor prognosis. Although patients benefited from AML-type induction chemotherapy followed by stem cell transplantation as postremission therapy, effective therapies, and prevention are needed. %+ Division of Pediatrics, The University of Texas MD Anderson Cancer Center, TX, USA. 8442 false false 11 Journal of pediatric hematology/oncology : official journal of the American Society of Pediatric Hematology/Oncology J Pediatr Hematol Oncol 803-811 2009-11-01 ppublish JOURNAL ARTICLE 19801947 In-Process 2009-11-07T07:18:23Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19801947 31 2009 Transforming growth factor (TGF)-beta is a central mediator in the progression of glomerulosclerosis, leading to accumulation of aberrant extracellular matrix (ECM) proteins and inappropriate expression of smooth muscle alpha-actin (alphaSMA) in the kidney. Previously, we reported that disrupting the cytoskeleton diminished TGF-beta-stimulated type I collagen accumulation in human mesangial cells. As cytoskeletal signaling molecules, including the Rho-family GTPases, have been implicated in fibrogenesis, we sought to determine the respective roles of RhoA and Rac1 in HMC collagen I expression. TGF-beta1 activated both RhoA and Rac1 within 5 min of treatment, and this activation was dependent on the kinase activity of the type I TGF-beta receptor (TbetaRI). TGF-beta1-stimulated induction of type I collagen mRNA expression and promoter activity was diminished by inhibiting Rac1 activity and was increased by a constitutively active Rac1 mutant, whereas inhibiting RhoA activity had no such effect. Rac1 activation required phosphatidylinositol-3-kinase (PI3K) activity. Further, the PI3K antagonist, LY294002, reduced TGF-beta1-stimulated COL1A2 promoter activity and Rac1 activation. It also partially blocked active Rac1-stimulated collagen promoter activity, suggesting that PI3K activity contributes to both TGF-beta activation of Rac1 and signal propagation downstream of Rac1. Thus, while both Rac1 and RhoA are rapidly activated in response to TGF-beta1 in human mesangial cells, only Rac1 activation enhances events that contribute to mesangial cell collagen expression, through a positive feedback loop involving PI3K. Key words: Kidney, Fibrosis, Rho-GTPase. Hubchak, S. C. Sparks, E. E. Hayashida, T. Schnaper, H. W. 0 2009-09-06T11:57:57Z 2009-09-02 2009-09-04 %0 Journal Article %A Hubchak, S. C. %A Sparks, E. E. %A Hayashida, T. %A Schnaper, H. W. %D 2009 %T Rac1 promotes TGF-beta-stimulated mesangial cell type I collagen expression through a PI3K/Akt-dependent mechanism. %J Am J Physiol Renal Physiol %V 297 %N 5 %P F1316-F1323 %M 19726546 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19726546 %X Transforming growth factor (TGF)-beta is a central mediator in the progression of glomerulosclerosis, leading to accumulation of aberrant extracellular matrix proteins and inappropriate expression of smooth muscle alpha-actin in the kidney. Previously, we reported that disrupting the cytoskeleton diminished TGF-beta-stimulated type I collagen accumulation in human mesangial cells. As cytoskeletal signaling molecules, including the Rho-family GTPases, have been implicated in fibrogenesis, we sought to determine the respective roles of RhoA and Rac1 in HMC collagen I expression. TGF-beta1 activated both RhoA and Rac1 within 5 min of treatment, and this activation was dependent on the kinase activity of the type I TGF-beta receptor. TGF-beta1-stimulated induction of type I collagen mRNA expression and promoter activity was diminished by inhibiting Rac1 activity and was increased by a constitutively active Rac1 mutant, whereas inhibiting RhoA activity had no such effect. Rac1 activation required phosphatidylinositol-3-kinase (PI3K) activity. Furthermore, the PI3K antagonist, LY294002, reduced TGF-beta1-stimulated COL1A2 promoter activity and Rac1 activation. It also partially blocked active Rac1-stimulated collagen promoter activity, suggesting that PI3K activity contributes to both TGF-beta activation of Rac1 and signal propagation downstream of Rac1. Thus, while both Rac1 and RhoA are rapidly activated in response to TGF-beta1 in human mesangial cells, only Rac1 activation enhances events that contribute to mesangial cell collagen expression, through a positive feedback loop involving PI3K. %K 1-Phosphatidylinositol 3-Kinase/*physiology %K Actins/analysis/metabolism %K Blotting, Northern %K Collagen Type I/*biosynthesis %K Fibrosis %K Humans %K Kidney/pathology %K Kidney Cortex/cytology %K Mesangial Cells/drug effects/*metabolism %K Oncogene Protein v-akt/*physiology %K Plasmids/genetics %K RNA/biosynthesis/isolation & purification %K Transfection %K Transforming Growth Factor beta/*pharmacology %K rac1 GTP-Binding Protein/*physiology %K rho GTP-Binding Proteins/metabolism %K rhoA GTP-Binding Protein/metabolism %+ Division of Kidney Diseases, Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. s-hubchak@northwestern.edu 5443 false false 5 American journal of physiology. Renal physiology Am J Physiol Renal Physiol 1-Phosphatidylinositol 3-Kinase/*physiology; Actins/analysis/metabolism; Blotting, Northern; Collagen Type I/*biosynthesis; Fibrosis; Humans; Kidney/pathology; Kidney Cortex/cytology; Mesangial Cells/drug effects/*metabolism; Oncogene Protein v-akt/*physiology; Plasmids/genetics; RNA/biosynthesis/isolation & purification; Transfection; Transforming Growth Factor beta/*pharmacology; rac1 GTP-Binding Protein/*physiology; rho GTP-Binding Proteins/metabolism; rhoA GTP-Binding Protein/metabolism F1316-F1323 2009-11-01 ppublish JOURNAL ARTICLE 19726546 MEDLINE 2009-11-18T07:16:46Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19726546 297 2009 Objectives Several drug resistance and secondary mutations have been described in HIV-1 viruses from patients undergoing antiretroviral therapy. In this study, we assessed the impact of the protease substitution T74S on the phenotype and on the replicative fitness in HIV-1 subtypes B and C. Methods HIV-1 molecular clones carrying subtype B or C proteases had these coding regions subjected to site-directed mutagenesis to include T74S alone or in combination with four known protease inhibitor (PI) primary drug resistance mutations. All clones were used in a phenotypic assay to evaluate their susceptibility to most commercially available PIs. The impact of T74S on virus fitness was also assessed for all viruses through head-to-head competitions and oligonucleotide ligation assays to measure the proportion of each virus in culture. Results Viruses of both subtypes carrying T74S did not have their susceptibility altered to any tested PI. Viruses with the four resistance mutations showed strong resistance to most PIs with fold changes ranging from 5 to 300 times compared with their wild-type counterparts. Surprisingly, the addition of T74S to the multiresistant clones restored their susceptibilities to indinavir and ritonavir and partially to lopinavir, close to those of wild-type viruses. Most 74S-containing viruses were more fit than their 74T counterparts. Conclusions Our results suggest that T74S is not a major drug resistance mutation, but it resensitizes multiresistant viruses to certain PIs. T74S is a bona fide accessory mutation, restoring fitness of multidrug-resistant viruses in both subtypes B and C. T74S should be further studied in clinical settings and considered in drug resistance interpretation algorithms. Soares, E. A. Santos, A. F. Gonzalez, L. M. Lalonde, M. S. Tebit, D. M. Tanuri, A. Arts, E. J. Soares, M. A. 0 2009-09-06T11:58:18Z 2009-08-26 2009-08-28 %0 Journal Article %A Soares, E. A. %A Santos, A. F. %A Gonzalez, L. M. %A Lalonde, M. S. %A Tebit, D. M. %A Tanuri, A. %A Arts, E. J. %A Soares, M. A. %D 2009 %T Mutation T74S in HIV-1 subtype B and C proteases resensitizes them to ritonavir and indinavir and confers fitness advantage. %J J Antimicrob Chemother %V 64 %N 5 %P 938-944 %M 19710076 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19710076 %X OBJECTIVES: Several drug resistance and secondary mutations have been described in HIV-1 viruses from patients undergoing antiretroviral therapy. In this study, we assessed the impact of the protease substitution T74S on the phenotype and on the replicative fitness in HIV-1 subtypes B and C. METHODS: HIV-1 molecular clones carrying subtype B or C proteases had these coding regions subjected to site-directed mutagenesis to include T74S alone or in combination with four known protease inhibitor (PI) primary drug resistance mutations. All clones were used in a phenotypic assay to evaluate their susceptibility to most commercially available PIs. The impact of T74S on virus fitness was also assessed for all viruses through head-to-head competitions and oligonucleotide ligation assays to measure the proportion of each virus in culture. RESULTS: Viruses of both subtypes carrying T74S did not have their susceptibility altered to any tested PI. Viruses with the four resistance mutations showed strong resistance to most PIs with fold changes ranging from 5 to 300 times compared with their wild-type counterparts. Surprisingly, the addition of T74S to the multiresistant clones restored their susceptibilities to indinavir and ritonavir and partially to lopinavir, close to those of wild-type viruses. Most 74S-containing viruses were more fit than their 74T counterparts. CONCLUSIONS: Our results suggest that T74S is not a major drug resistance mutation, but it resensitizes multiresistant viruses to certain PIs. T74S is a bona fide accessory mutation, restoring fitness of multidrug-resistant viruses in both subtypes B and C. T74S should be further studied in clinical settings and considered in drug resistance interpretation algorithms. %+ Departamento de Genetica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil. 5728 false false 5 The Journal of antimicrobial chemotherapy J Antimicrob Chemother 938-944 2009-11-01 ppublish JOURNAL ARTICLE 19710076 In-Process 2009-10-15T06:14:48Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19710076 64 2009 Trypanosoma congolense is one of the most economically important pathogens of livestock in Africa. Culture-derived parasites of each of the three main insect stages of the T. congolense life cycle, i.e., the procyclic, epimastigote and metacyclic stages, and bloodstream stage parasites isolated from infected mice, were used to construct stage-specific cDNA libraries and expressed sequence tags (ESTs or cDNA clones) in each library were sequenced. Thirteen EST clusters encoding different variant surface glycoproteins (VSGs) were detected in the metacyclic library and 26 VSG EST clusters were found in the bloodstream library, 6 of which are shared by the metacyclic library. Rare VSG ESTs are present in the epimastigote library, and none were detected in the procyclic library. ESTs encoding enzymes that catalyze oxidative phosphorylation and amino acid metabolism are about twice as abundant in the procyclic and epimastigote stages as in the metacyclic and bloodstream stages. In contrast, ESTs encoding enzymes involved in glycolysis, the citric acid cycle and nucleotide metabolism are about the same in all four developmental stages. Cysteine proteases, kinases and phosphatases are the most abundant enzyme groups represented by the ESTs. All four libraries contain T. congolense-specific expressed sequences not present in the Trypanosoma brucei and Trypanosoma cruzi genomes. Normalized cDNA libraries were constructed from the metacyclic and bloodstream stages, and found to be further enriched for T. congolense-specific ESTs. Given that cultured T. congolense offers an experimental advantage over other African trypanosome species, these ESTs provide a basis for further investigation of the molecular properties of these four developmental stages, especially the epimastigote and metacyclic stages for which it is difficult to obtain large quantities of organisms. The T. congolense EST databases are available at: http://www.sanger.ac.uk/Projects/T_congolense/EST_index.shtml. The sequence data have been submitted to EMBL under the following accession numbers: FN263376-FN292969. Helm, J. R. Hertz-Fowler, C. Aslett, M. Berriman, M. Sanders, M. Quail, M. A. Soares, M. B. Bonaldo, M. F. Sakurai, T. Inoue, N. Donelson, J. E. 0 2009-09-06T11:58:18Z 2009-06-25 2009-06-30 %0 Journal Article %A Helm, J. R. %A Hertz-Fowler, C. %A Aslett, M. %A Berriman, M. %A Sanders, M. %A Quail, M. A. %A Soares, M. B. %A Bonaldo, M. F. %A Sakurai, T. %A Inoue, N. %A Donelson, J. E. %D 2009 %T Analysis of expressed sequence tags from the four main developmental stages of Trypanosoma congolense. %J Mol Biochem Parasitol %V 168 %N 1 %P 34-42 %M 19559733 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19559733 %X Trypanosoma congolense is one of the most economically important pathogens of livestock in Africa. Culture-derived parasites of each of the three main insect stages of the T. congolense life cycle, i.e., the procyclic, epimastigote and metacyclic stages, and bloodstream stage parasites isolated from infected mice, were used to construct stage-specific cDNA libraries and expressed sequence tags (ESTs or cDNA clones) in each library were sequenced. Thirteen EST clusters encoding different variant surface glycoproteins (VSGs) were detected in the metacyclic library and 26 VSG EST clusters were found in the bloodstream library, 6 of which are shared by the metacyclic library. Rare VSG ESTs are present in the epimastigote library, and none were detected in the procyclic library. ESTs encoding enzymes that catalyze oxidative phosphorylation and amino acid metabolism are about twice as abundant in the procyclic and epimastigote stages as in the metacyclic and bloodstream stages. In contrast, ESTs encoding enzymes involved in glycolysis, the citric acid cycle and nucleotide metabolism are about the same in all four developmental stages. Cysteine proteases, kinases and phosphatases are the most abundant enzyme groups represented by the ESTs. All four libraries contain T. congolense-specific expressed sequences not present in the Trypanosoma brucei and Trypanosoma cruzi genomes. Normalized cDNA libraries were constructed from the metacyclic and bloodstream stages, and found to be further enriched for T. congolense-specific ESTs. Given that cultured T. congolense offers an experimental advantage over other African trypanosome species, these ESTs provide a basis for further investigation of the molecular properties of these four developmental stages, especially the epimastigote and metacyclic stages for which it is difficult to obtain large quantities of organisms. The T. congolense EST databases are available at: http://www.sanger.ac.uk/Projects/T_congolense/EST_index.shtml. The sequence data have been submitted to EMBL under the following accession numbers: FN263376-FN292969. %K Animals %K DNA, Protozoan/genetics %K *Expressed Sequence Tags %K *Gene Expression Profiling %K *Gene Library %K Genes, Protozoan %K Mice %K Multigene Family %K Trypanosoma congolense/*genetics/*growth & development %+ Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA. Helm, Jared R Hertz-Fowler, Christiane Aslett, Martin Berriman, Matthew Sanders, Mandy Quail, Michael A Soares, Marcelo B Bonaldo, Maria F Sakurai, Tatsuya Inoue, Noboru Donelson, John E 5730 false false 1 Molecular and biochemical parasitology Mol Biochem Parasitol Animals; DNA, Protozoan/genetics; *Expressed Sequence Tags; *Gene Expression Profiling; *Gene Library; Genes, Protozoan; Mice; Multigene Family; Trypanosoma congolense/*genetics/*growth & development 34-42 2009-11-01 ppublish Journal Article 19559733 MEDLINE 2009-10-17T06:19:28Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19559733 168 2009 We report the case of an adult patient with pineoblastoma (PBL) who had a complete radiographic response following treatment with vorinostat and retinoic acid. This regimen was used to treat bulky residual tumor that persisted despite radiation therapy (RT) and two cycles of cytotoxic chemotherapy. Vorinostat and retinoic acid were chosen as an alternative to cytotoxic chemotherapy, which our patient was unable to tolerate, based on preclinical data suggesting efficacy of this combination. MRI demonstrated a complete response to this regimen, which continues to remain stable without evidence of recurrence. Deboer, R. Batjer, H. Marymont, M. Goldman, S. Walker, M. Gottardi-Littell, N. Raizer, J. 0 2009-09-06T11:55:17Z 2009-06-09 2009-06-10 %0 Journal Article %A DeBoer, R. %A Batjer, H. %A Marymont, M. %A Goldman, S. %A Walker, M. %A Gottardi-Littell, N. %A Raizer, J. %D 2009 %T Response of an adult patient with pineoblastoma to vorinostat and retinoic acid. %J J Neurooncol %V 95 %N 2 %P 289-292 %M 19506816 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19506816 %X We report the case of an adult patient with pineoblastoma (PBL) who had a complete radiographic response following treatment with vorinostat and retinoic acid. This regimen was used to treat bulky residual tumor that persisted despite radiation therapy (RT) and two cycles of cytotoxic chemotherapy. Vorinostat and retinoic acid were chosen as an alternative to cytotoxic chemotherapy, which our patient was unable to tolerate, based on preclinical data suggesting efficacy of this combination. MRI demonstrated a complete response to this regimen, which continues to remain stable without evidence of recurrence. %+ Feinberg School of Medicine, Northwestern University, 710 N Lake Shore Drive, Abbott Hall, Room 1123, Chicago, IL 60611, USA. becky.deboer@gmail.com 1683 false false 2 Journal of neuro-oncology J Neurooncol 289-292 2009-11-01 ppublish JOURNAL ARTICLE 19506816 In-Process 2009-10-09T06:13:29Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19506816 95 2009 We have previously found that certain stem cells that are derived from rat blastocysts and named extraembryonic endoderm precursor (XEN-P) cells, show a unique molecular signature sharing some of the characteristics of embryonic stem cells (ES), trophoblast stem cells (TS) and extraembryonic endoderm stem cells (XEN). These XEN-P cells are positive for AP, SSEA1, Oct4 and Rex1 markers similar to ES cells and also express signature markers of TS - eomesodermin (Eomes) and XEN - Gata6. Here we show that these cells integrate not only into the visceral and parietal extraembryonic endoderm lineages as observed before, but also into the inner cell mass (ICM), the primitive endoderm, and the polar and mural trophectoderm of cultured embryos. In addition, we find that the XEN-P cells colonize yolk sac and contribute to trophoblast lineages of post-implantation embryos following transfer to surrogate mothers. We also find that the XEN-P cell culture propagates by shedding cell clusters into the media in addition to typical expansion of colonies. Interestingly, the cell cultures exist as mixed populations of two interconvertible phenotypes of flat and round cells with preferential expression of stem cell markers Oct 4 and SSEA1 in round cells. We believe these cells represent a metastable stage during ICM cellular segregation. These results are important to developing hypotheses of cell fate plasticity in the ICM and provide a model for the study of development and differentiation along the extraembryonic lineages. Galat, V. Binas, B. Iannaccone, S. Postovit, L. M. Debeb, B. Iannaccone, P. 0 2009-09-06T11:55:40Z 2009-06-02 %0 Journal Article %A Galat, V. %A Binas, B. %A Iannaccone, S. %A Postovit, L. M. %A Debeb, B. G. %A Iannaccone, P. %D 2009 %T Developmental potential of rat extraembryonic stem cells. %J Stem Cells Dev %V 18 %N 9 %P 1309-1318 %M 19480599 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19480599 %X We have previously found that certain stem cells that are derived from rat blastocysts and named extraembryonic endoderm precursor (XEN-P) cells show a unique molecular signature sharing some of the characteristics of embryonic stem cells (ES), trophoblast stem cells (TS), and extraembryonic endoderm stem cells (XEN). These XEN-P cells are positive for AP, SSEA1, Oct4, and Rex1 markers similar to ES cells and also express signature markers of TS-eomesodermin (Eomes) and XEN-Gata6. Here we show that these cells integrate into the visceral and parietal extraembryonic endoderm lineages as well as into the inner cell mass (ICM), the primitive endoderm, and the polar and mural trophectoderm (TE) of cultured embryos. In addition, we find that the XEN-P cells colonize yolk sac and contribute to trophoblast lineages of postimplantation embryos following transfer to surrogate mothers. We also find that the XEN-P cell culture propagates by shedding cell clusters into the media in addition to typical expansion of colonies. Interestingly, the cell cultures exist as mixed populations of two interconvertible phenotypes of flat and round cells with preferential expression of stem cell markers Oct4 and SSEA1 in round cells. We believe these cells represent a metastable stage during ICM cellular segregation. These results are important for developing hypotheses of cell fate plasticity in the ICM and provide a model for the study of development and differentiation along the extraembryonic lineages. %+ Developmental Biology Program, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60614, USA. v-galat@northwestern.edu 2411 false false 9 Stem cells and development Stem Cells Dev 1309-1318 2009-11-01 ppublish JOURNAL ARTICLE 19480599 In-Process 2009-11-17T07:20:54Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19480599 18 2009 The cystic fibrosis transmembrane conductance regulator (CFTR) gene is driven by a promoter that cannot alone account for the temporal and tissue-specific regulation of the gene. This has led to the search for additional regulatory elements that cooperate with the basal promoter to achieve coordinated expression. We previously identified two alternative upstream exons of the gene that were mutually exclusive with the first exon and one of which showed temporal regulation in the human and sheep lung. We now demonstrate that this alternative splice product generates a stable protein, which initiates translation at an ATG in exon 4 and thus lacks the N-terminus of CFTR. The other splice variant inhibits translation of the protein. In a search for the promoter utilized by the upstream exons, we identified a novel element that contributes to the activity of the basal CFTR promoter in airway epithelial cells, but does not function independently. Finally, we demonstrated that in primary airway cells, skin fibroblasts, and both airway and intestinal cell lines, the CFTR promoter is unmethylated, irrespective of CFTR expression status. Thus methylation is not the main cause of inactivation of CFTR transcription. Lewandowska, M. A. Costa, F. F. Bischof, J. M. Williams, S. H. Soares, M. B. Harris, A. 0 2009-10-28T06:22:33Z 2009-10-23 2009-10-27 %0 Journal Article %A Lewandowska, M. A. %A Costa, F. F. %A Bischof, J. M. %A Williams, S. H. %A Soares, M. B. %A Harris, A. %D 2009 %T Multiple Mechanisms Influence Regulation of the CFTR Gene Promoter. %J Am J Respir Cell Mol Biol %M 19855085 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19855085 %X The cystic fibrosis transmembrane conductance regulator (CFTR) gene is driven by a promoter that cannot alone account for the temporal and tissue-specific regulation of the gene. This has led to the search for additional regulatory elements that cooperate with the basal promoter to achieve coordinated expression. We previously identified two alternative upstream exons of the gene that were mutually exclusive with the first exon and one of which showed temporal regulation in the human and sheep lung. We now demonstrate that this alternative splice product generates a stable protein, which initiates translation at an ATG in exon 4 and thus lacks the N-terminus of CFTR. The other splice variant inhibits translation of the protein. In a search for the promoter utilized by the upstream exons, we identified a novel element that contributes to the activity of the basal CFTR promoter in airway epithelial cells, but does not function independently. Finally, we demonstrated that in primary airway cells, skin fibroblasts, and both airway and intestinal cell lines, the CFTR promoter is unmethylated, irrespective of CFTR expression status. Thus methylation is not the main cause of inactivation of CFTR transcription. %+ Children's Memorial Research Center, Pediatrics, Northwestern University, Feinberg School of Medicine, Human Molecular Genetic Program, Chicago, Illinois, United States. 8531 false false American journal of respiratory cell and molecular biology Am J Respir Cell Mol Biol 2009-10-23 aheadofprint JOURNAL ARTICLE 19855085 Publisher 2009-10-28T06:22:33Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19855085 2009 A fluorescent chemodosimeter for cysteine detection and bio-imaging has been described which displays an excellent selectivity for cysteine over homocysteine, glutathione and other amino acids. Li, H. Fan, J. Wang, J. Tian, M. Du, J. Sun, S. Sun, P. Peng, X. 0 2009-10-01T06:13:47Z 2009-08-20 2009-09-30 %0 Journal Article %A Li, H. %A Fan, J. %A Wang, J. %A Tian, M. %A Du, J. %A Sun, S. %A Sun, P. %A Peng, X. %D 2009 %T A fluorescent chemodosimeter specific for cysteine: effective discrimination of cysteine from homocysteine. %J Chem Commun (Camb) %N 39 %P 5904-5906 %M 19787136 %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19787136 %X A fluorescent chemodosimeter for cysteine detection and bio-imaging has been described which displays an excellent selectivity for cysteine over homocysteine, glutathione and other amino acids. %+ State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China. Li, Honglin Fan, Jiangli Wang, Jingyun Tian, Maozhong Du, Jianjun Sun, Shiguo Sun, Pingping Peng, Xiaojun 8420 false false 39 Chemical communications (Cambridge, England) Chem Commun (Camb) 5904-5906 2009-10-21 ppublish Journal Article 19787136 In-Process 2009-10-01T06:13:47Z http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19787136 2009