50 results on '"Bollag R"'
Search Results
2. Comparative analysis of the cellular profile and architecture of metastatic and non-metastatic lymph nodes
- Author
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Booth, J.R., primary, Guo, G., additional, Myint, C., additional, Patel, N., additional, Bollag, R., additional, Albergotti, W., additional, Groves, M., additional, Byrd, J.K., additional, and Cui, Y., additional
- Published
- 2020
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3. Osteoblast-Derived Cells Express Functional Glucose-Dependent Insulinotropic Peptide Receptors*
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Bollag, R J, Zhong, Q, Phillips, P, Min, L, Zhong, L, Cameron, R, Mulloy, A L, Rasmussen, H, Qin, F, Ding, K H, and Isales, C M
- Published
- 2000
4. Multiple melanocortin receptors are expressed in bone cells
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ZHONG, Q, primary, SRIDHAR, S, additional, RUAN, L, additional, DING, K, additional, XIE, D, additional, INSOGNA, K, additional, KANG, B, additional, XU, J, additional, BOLLAG, R, additional, and ISALES, C, additional
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- 2005
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5. Spontaneous and restriction enzyme-induced chromosomal recombination in mammalian cells.
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Godwin, A R, primary, Bollag, R J, additional, Christie, D M, additional, and Liskay, R M, additional
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- 1994
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6. Formation of heteroduplex DNA during mammalian intrachromosomal gene conversion.
- Author
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Bollag, R J, primary, Elwood, D R, additional, Tobin, E D, additional, Godwin, A R, additional, and Liskay, R M, additional
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- 1992
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7. Direct-repeat analysis of chromatid interactions during intrachromosomal recombination in mouse cells
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Bollag, R J, primary and Liskay, R M, additional
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- 1991
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8. Functional analysis of glucose-dependent insulinotropic polypeptide fusion proteins
- Author
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Ding, K. H., Isalesab, C. M., Zhong, Q., and Bollag, R. J.
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- 2001
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9. Glucose-dependent insulinotropic peptide is an integrative hormone with osteotropic effects
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Bollag, R. J., Zhong, Q., Ding, K. H., Phillips, P., Zhong, L., Qin, F., Cranford, J., Mulloy, A. L., Cameron, R., and Isales, C. M.
- Published
- 2001
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10. 1,25-Dihydroxyvitamin D3, phospholipase D and protein kinase C in keratinocyte differentiation
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Bollag, W. Bollinger and Bollag, R. J.
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- 2001
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11. Microarray analysis of Tbx2-directed gene expression: a possible role in osteogenesis
- Author
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Chen, J. R., Zhong, Q., Wang, J., Cameron, R. S., Borke, J. L., Isales, C. M., and Bollag, R. J.
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- 2001
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12. Glucose-dependent insulinotropic peptide signaling pathways in endothelial cells
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Zhong, Q., Bollag, R. J., Dransfield, D. T., Gasalla-Herraiz, J., Ding, K. H., Min, L., and Isales, C. M.
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- 2000
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13. 1,25-dihydroxyvitamin D3 induces phospholipase D-1 expression in primary mouse epidermal keratinocytes.
- Author
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Griner, R D, Qin, F, Jung, E, Sue-Ling, C K, Crawford, K B, Mann-Blakeney, R, Bollag, R J, and Bollag, W B
- Abstract
The hormone 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) elicits the programmed pattern of differentiation in epidermal keratinocytes. Based on data indicating a potential role of phospholipase D (PLD) in mediating keratinocyte differentiation, we investigated the effect of 1,25(OH)2D3 on PLD expression. A 24-h exposure to 1, 25(OH)2D3 stimulated PLD-1, but not PLD-2, mRNA expression. This 1, 25(OH)2D3-enhanced expression was accompanied by increased total PLD and PLD-1 activity. Time course studies indicated that 1,25(OH)2D3 induced PLD-1 expression by 8 h, with a maximal increase at 20-24 h. Exposure to 1,25(OH)2D3 inhibited proliferation over the same time period with similar kinetics. Expression of the early (spinous) differentiation marker keratin 1 decreased in response to 1, 25(OH)2D3 over 12-24 h. Treatment with 1,25(OH)2D3 enhanced the activity of transglutaminase, a late (granular) differentiation marker, by 12 h with a maximal increase after 24 h. In situ hybridization studies demonstrated that the highest levels of PLD-1 expression are in the more differentiated (spinous and granular) layers of the epidermis, with little expression in basal keratinocytes. Our results suggest a role for PLD expression/activity during keratinocyte differentiation.
- Published
- 1999
14. Conservative intrachromosomal recombination between inverted repeats in mouse cells: association between reciprocal exchange and gene conversion.
- Author
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Bollag, R J, primary and Liskay, R M, additional
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- 1988
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15. S062 Nuclear, Cytoplasmic Expression of Galectin-3 Is Associated With ß-Catenin/Wnt-Pathway Activation in Thyroid Carcinoma
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Weinberger, P. M., Adam, B., Gourin, C. G., Moretz III, W. H., Bollag, R. J., Lee, J. R., and Terris, D. J.
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- 2006
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16. Primary tumor-induced immunity eradicates disseminated tumor cells in syngeneic mouse model
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John K. Cowell, Muthusamy Thangaraju, Khaled A. Hassan, Mustafa Guzel, Hasan Korkaya, Esteban Celis, Maria Ouzounova, Ali S. Arbab, Raziye Piranlioglu, Eunmi Lee, Daniela Marasco, Ahmed Chadli, Max S. Wicha, Roni J. Bollag, Alicia Vinyard, Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Piranlioglu, Raziye, Lee, EunMi, Bollag, Roni J., Vinyard, Alicia H., Arbab, Ali S., Cowell, John K., Thangaraju, Muthushamy, Chadli, Ahmed, Celis, Esteban, Korkaya, Hasan Augusta Univ, Dept Biochem & Mol Biol, Georgia Canc Ctr, 1410 Laney Walker Blvd CN2136, Augusta, GA 30912 USA, Ouzounova, Maria Canc Res Ctr Lyon, 28 Rue Laennec, F-69008 Lyon, France, Marasco, Daniela Univ Naples Federico II, Dept Pharm, I-80134 Naples, Italy, Guzel, Mustafa Medipol Univ, REMER, Kavacik Mah Ekinciler Cad 19, TR-34810 Istanbul, Turkey, Hassan, Khaled A., Wicha, Max S. Univ Michigan, Comprehens Canc Ctr, 1500 E Med Ctr Dr, Ann Arbor, MI 48109 USA, Ouzounova, Maria -- 0000-0001-8934-3461, Korkaya, Hasan -- 0000-0002-0719-5862, Guzel, Prof. Mustafa -- 0000-0002-1423-0435, Bollag, Roni -- 0000-0003-4963-6619, Piranlioglu, Raziye -- 0000-0003-2450-3887, Piranlioglu, R., Lee, E. M., Ouzounova, M., Bollag, R. J., Vinyard, A. H., Arbab, A. S., Marasco, D., Guzel, M., Cowell, J. K., Thangaraju, M., Chadli, A., Hassan, K. A., Wicha, M. S., Celis, E., and Korkaya, H.
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0301 basic medicine ,Adoptive cell transfer ,Disseminated Tumor Cells (DTC) ,[SDV]Life Sciences [q-bio] ,General Physics and Astronomy ,Triple Negative Breast Neoplasms ,02 engineering and technology ,CD8-Positive T-Lymphocytes ,Metastasis ,Mice ,Neoplasms ,Disease Models, Animal, Biological ,Neoplasm Metastasis ,lcsh:Science ,ComputingMilieux_MISCELLANEOUS ,Multidisciplinary ,Tumor ,021001 nanoscience & nanotechnology ,Primary tumor ,3. Good health ,Neoplasm Metastasi ,Survival Analysi ,0210 nano-technology ,Primary ,Tail ,Science ,Cells ,Models, Biological ,Article ,General Biochemistry, Genetics and Molecular Biology ,Veins ,03 medical and health sciences ,Triple Negative Breast ,Immunity ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Neoplasm Invasiveness ,Cell Proliferation ,Cell growth ,business.industry ,General Chemistry ,CD8-Positive T-Lymphocyte ,medicine.disease ,Survival Analysis ,Lymphocyte Subsets ,Disease Models, Animal ,030104 developmental biology ,Tumor progression ,Cell culture ,Cancer research ,lcsh:Q ,business ,CD8 - Abstract
Although clinically apparent metastasis is associated with late stages of cancer development, micro-metastatic dissemination may be an early event. However, the fate of these early disseminated tumor cells (DTC) remains elusive. We show that despite their capacity to disseminate into secondary organs, 4T1 tumor models develop overt metastasis while EMT6-tumor bearing mice clear DTCs shed from primary tumors as well as those introduced by intravenous (IV) injection. Following the surgical resection of primary EMT6 tumors, mice do not develop detectable metastasis and reject IV-injected tumor cells. In contrast, these cells readily grow and metastasize in immuno-deficient athymic or Rag2−/− mice, an effect mimicked by CD8+ T-cell depletion in immunocompetent mice. Furthermore, recombinant G-CSF or adoptive transfer of granulocytic-MDSCs isolated from 4T1 tumor-bearing mice, induce metastasis by suppressing CD8+ T-cells in EMT6-primed mice. Our studies support the concept of immune surveillance providing molecular insights into the immune mechanisms during tumor progression., Dissemination of tumor cells from the primary site is an early event. Here, the authors show that the early disseminated tumor cells are actively cleared by the host cytotoxic T lymphocytes induced by the primary tumor and that infiltration of granulocytic myeloid-derived suppressor cells counteracts such immune protection and allow metastasis development.
- Published
- 2019
17. Simple virus-free mouse models of COVID-19 pathologies and oral therapeutic intervention.
- Author
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Zhu H, Sharma AK, Aguilar K, Boghani F, Sarcan S, George M, Ramesh J, Van Der Eerden J, Panda CS, Lopez A, Zhi W, Bollag R, Patel N, Klein K, White J, Thangaraju M, Lokeshwar BL, Singh N, and Lokeshwar VB
- Abstract
The paucity of preclinical models that recapitulate COVID-19 pathology without requiring SARS-COV-2 adaptation and humanized/transgenic mice limits research into new therapeutics against the frequently emerging variants-of-concern. We developed virus-free models by C57BL/6 mice receiving oropharyngeal instillations of a SARS-COV-2 ribo-oligonucleotide common in all variants or specific to Delta/Omicron variants, concurrently with low-dose bleomycin. Mice developed COVID-19-like lung pathologies including ground-glass opacities, interstitial fibrosis, congested alveoli, and became moribund. Lung tissues from these mice and bronchoalveolar lavage and lung tissues from patients with COVID-19 showed elevated levels of hyaluronic acid (HA), HA-family members, an inflammatory signature, and immune cell infiltration. 4-methylumbelliferone (4-MU), an oral drug for biliary-spasm treatment, inhibits HA-synthesis. At the human equivalent dose, 4-MU prevented/inhibited COVID-19-like pathologies and long-term morbidity; 4-MU and metabolites accumulated in mice lungs. Therefore, these versatile SARS-COV-2 ribo-oligonucleotide oropharyngeal models recapitulate COVID-19 pathology, with HA as its critical mediator and 4-MU as a potential therapeutic for COVID-19., Competing Interests: None., (© 2024 The Authors.)
- Published
- 2024
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18. Indomethacin-induced oxidative stress enhances death receptor 5 signaling and sensitizes tumor cells to adoptive T-cell therapy.
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Aboelella NS, Brandle C, Okoko O, Gazi MY, Ding ZC, Xu H, Gorman G, Bollag R, Davila ML, Bryan LJ, Munn DH, Piazza GA, and Zhou G
- Subjects
- Animals, Anti-Inflammatory Agents, Non-Steroidal pharmacology, Cell- and Tissue-Based Therapy, Humans, Mice, Neoplasm Recurrence, Local, Oxidative Stress, TNF-Related Apoptosis-Inducing Ligand, Indomethacin pharmacology, Receptors, TNF-Related Apoptosis-Inducing Ligand
- Abstract
Background: Adoptive cell therapy (ACT) using genetically modified T cells has evolved into a promising treatment option for patients with cancer. However, even for the best-studied and clinically validated CD19-targeted chimeric antigen receptor (CAR) T-cell therapy, many patients face the challenge of lack of response or occurrence of relapse. There is increasing need to improve the efficacy of ACT so that durable, curative outcomes can be achieved in a broad patient population., Methods: Here, we investigated the impact of indomethacin (indo), a non-steroidal anti-inflammatory drug (NSAID), on the efficacy of ACT in multiple preclinical models. Mice with established B-cell lymphoma received various combinations of preconditioning chemotherapy, infusion of suboptimal dose of tumor-reactive T cells, and indo administration. Donor T cells used in the ACT models included CD4
+ T cells expressing a tumor-specific T cell receptor (TCR) and T cells engineered to express CD19CAR. Mice were monitored for tumor growth and survival. The effects of indo on donor T cell phenotype and function were evaluated. The molecular mechanisms by which indo may influence the outcome of ACT were investigated., Results: ACT coupled with indo administration led to improved tumor growth control and prolonged mouse survival. Indo did not affect the activation status and tumor infiltration of the donor T cells. Moreover, the beneficial effect of indo in ACT did not rely on its inhibitory effect on the immunosuppressive cyclooxygenase 2 (COX2)/prostaglandin E2 (PGE2) axis. Instead, indo-induced oxidative stress boosted the expression of death receptor 5 (DR5) in tumor cells, rendering them susceptible to donor T cells expressing TNF-related apoptosis-inducing ligand (TRAIL). Furthermore, the ACT-potentiating effect of indo was diminished against DR5-deficient tumors, but was amplified by donor T cells engineered to overexpress TRAIL., Conclusion: Our results demonstrate that the pro-oxidative property of indo can be exploited to enhance death receptor signaling in cancer cells, providing rationale for combining indo with genetically modified T cells to intensify tumor cell killing through the TRAIL-DR5 axis. These findings implicate indo administration, and potentially similar use of other NSAIDs, as a readily applicable and cost-effective approach to augment the efficacy of ACT., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)- Published
- 2022
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19. Chemoenzymatic modular assembly of O-GalNAc glycans for functional glycomics.
- Author
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Wang S, Chen C, Gadi MR, Saikam V, Liu D, Zhu H, Bollag R, Liu K, Chen X, Wang F, Wang PG, Ling P, Guan W, and Li L
- Subjects
- Carbohydrates, Carrier Proteins metabolism, Epitopes, Glycosylation, Humans, Microarray Analysis, Glycomics, Mucins chemistry, Mucins metabolism, Polysaccharides chemistry, Polysaccharides metabolism
- Abstract
O-GalNAc glycans (or mucin O-glycans) play pivotal roles in diverse biological and pathological processes, including tumor growth and progression. Structurally defined O-GalNAc glycans are essential for functional studies but synthetic challenges and their inherent structural diversity and complexity have limited access to these compounds. Herein, we report an efficient and robust chemoenzymatic modular assembly (CEMA) strategy to construct structurally diverse O-GalNAc glycans. The key to this strategy is the convergent assembly of O-GalNAc cores 1-4 and 6 from three chemical building blocks, followed by enzymatic diversification of the cores by 13 well-tailored enzyme modules. A total of 83 O-GalNAc glycans presenting various natural glycan epitopes are obtained and used to generate a unique synthetic mucin O-glycan microarray. Binding specificities of glycan-binding proteins (GBPs) including plant lectins and selected anti-glycan antibodies towards these O-GalNAc glycans are revealed by this microarray, promoting their applicability in functional O-glycomics. Serum samples from colorectal cancer patients and healthy controls are assayed using the array reveal higher bindings towards less common cores 3, 4, and 6 than abundant cores 1 and 2, providing insights into O-GalNAc glycan structure-activity relationships.
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- 2021
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20. Engaging Pathology Residents in Scholarly Activities.
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Singh G, Bollag R, Kolhe R, McMullen A, and Savage NM
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- Humans, Mentors, Internship and Residency
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- 2021
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21. Retrospective Validation of a 168-Gene Expression Signature for Glioma Classification on a Single Molecule Counting Platform.
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Tran PMH, Tran LKH, Satter KB, Purohit S, Nechtman J, Hopkins DI, Dos Santos B, Bollag R, Kolhe R, Sharma S, and She JX
- Abstract
Gene expression profiling has been shown to be comparable to other molecular methods for glioma classification. We sought to validate a gene-expression based glioma classification method. Formalin-fixed paraffin embedded tissue and flash frozen tissue collected at the Augusta University (AU) Pathology Department between 2000-2019 were identified and 2 mm cores were taken. The RNA was extracted from these cores after deparaffinization and bead homogenization. One hundred sixty-eight genes were evaluated in the RNA samples on the nCounter instrument. Forty-eight gliomas were classified using a supervised learning algorithm trained by using data from The Cancer Genome Atlas. An ensemble of 1000 linear support vector models classified 30 glioma samples into TP1 with classification confidence of 0.99. Glioma patients in TP1 group have a poorer survival (HR (95% CI) = 4.5 (1.3-15.4), p = 0.005) with median survival time of 12.1 months, compared to non-TP1 groups. Network analysis revealed that cell cycle genes play an important role in distinguishing TP1 from non-TP1 cases and that these genes may play an important role in glioma survival. This could be a good clinical pipeline for molecular classification of gliomas.
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- 2021
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22. Inclusion and Diversity in Pathology Residency Training.
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Savage NM, Bollag R, and Singh G
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- Humans, Internship and Residency
- Published
- 2021
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23. Comparative analysis of transcriptomic profile, histology, and IDH mutation for classification of gliomas.
- Author
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Tran PMH, Tran LKH, Nechtman J, Dos Santos B, Purohit S, Satter KB, Dun B, Kolhe R, Sharma S, Bollag R, and She JX
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- Astrocytoma genetics, Astrocytoma pathology, Biomarkers, Tumor genetics, Brain Neoplasms pathology, Cell Proliferation genetics, DNA Methylation genetics, Gene Expression genetics, Gene Expression Profiling methods, Glioma pathology, Humans, Neurons pathology, Prognosis, Brain Neoplasms genetics, Glioma genetics, Isocitrate Dehydrogenase genetics, Mutation genetics, Transcriptome genetics
- Abstract
Gliomas are currently classified through integration of histology and mutation information, with new developments in DNA methylation classification. However, discrepancies exist amongst the major classification methods. This study sought to compare transcriptome-based classification to the established methods. RNAseq and microarray data were obtained for 1032 gliomas from the TCGA and 395 gliomas from REMBRANDT. Data were analyzed using unsupervised and supervised learning and other statistical methods. Global transcriptomic profiles defined four transcriptomic glioma subgroups with 91.4% concordance with the WHO-defined mutation subtypes. Using these subgroups, 168 genes were selected for the development of 1000 linear support vector classifiers (LSVC). Based on plurality voting of 1000 LSVC, the final ensemble classifier confidently classified all but 17 TCGA gliomas to one of the four transcriptomic profile (TP) groups. The classifier was validated using a gene expression microarray dataset. TP1 cases include IDHwt, glioblastoma high immune infiltration and cellular proliferation and poor survival prognosis. TP2a is characterized as IDHmut-codel, oligodendrogliomas with high tumor purity. TP2b tissue is mostly composed of neurons and few infiltrating malignant cells. TP3 exhibit increased NOTCH signaling, are astrocytoma and IDHmut-non-codel. TP groups are highly concordant with both WHO integrated histology and mutation classification as well as methylation-based classification of gliomas. Transcriptomic profiling provides a robust and objective method to classify gliomas with high agreement to the current WHO guidelines and may provide additional survival prediction to the current methods.
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- 2020
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24. Quantification by Ultrafiltration and Immunofixation Electrophoresis Testing for Monoclonal Serum Free Light Chains.
- Author
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Singh G and Bollag R
- Subjects
- Biomarkers, Humans, Immunoglobulin kappa-Chains blood, Immunoglobulin lambda-Chains blood, Multiple Myeloma blood, Multiple Myeloma diagnosis, Sensitivity and Specificity, Antibodies, Monoclonal blood, Immunoelectrophoresis methods, Immunoelectrophoresis standards, Immunoglobulin Light Chains blood, Ultrafiltration methods, Ultrafiltration standards
- Abstract
Objective: Measurement of monoclonal immunoglobulins is a reliable estimate of the plasma cell tumor mass. About 15% of plasma cell myelomas secrete light chains only. The concentration of serum free light chains is insufficient evidence of the monoclonal light chain burden. A sensitive quantitative estimate of serum free monoclonal light chains could be useful for monitoring patients with light chain myeloma. We describe such an assay that does not require mass-spectrometry equipment or expertise., Methods: Serum specimens from patients with known light chain myelomas and controls were subjected to ultrafiltration through a membrane with pore size of 50 kDa. The filtrate was concentrated and tested by immunofixation electrophoresis. The relative area under the monoclonal peak, compared to that of the total involved light chain composition, was estimated by densitometric scanning of immunofixation gels. The proportion of the area occupied by the monoclonal peak in representative densitometric scans was used to arrive at the total serum concentration of the monoclonal serum free light chains., Results: Using an ultracentrifugation and concentration process, monoclonal serum free light chains were detectable, along with polyclonal light chains, in all 10 patients with active light chain myelomas. Monoclonal light chains were identified in serum specimens that did not reveal monoclonal light chains by conventional immunofixation electrophoresis. The limit of detection by this method was 1.0 mg/L of monoclonal serum free light chains., Conclusion: The method described here is simple enough to be implemented in academic medical center clinical laboratories and does not require special reagents, equipment, or expertise. Even though urine examination is the preferred method for the diagnosis of light chain plasma cell myelomas, measurement of the concentration of serum free light chains provides a convenient, albeit inadequate, way to monitor the course of disease. The method described here allows effective electrophoretic differentiation of monoclonal serum free light chain from polyclonal serum free light chains and provides a quantitation of the monoclonal serum free light chains in monitoring light chain monoclonal gammopathies., (© American Society for Clinical Pathology 2020. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
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- 2020
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25. Promotion of epithelial hyperplasia by interleukin-8-CXCR axis in human prostate.
- Author
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Smith DK, Hasanali SL, Wang J, Kallifatidis G, Morera DS, Jordan AR, Terris MK, Klaassen Z, Bollag R, Lokeshwar VB, and Lokeshwar BL
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- Cell Growth Processes drug effects, Cell Line, Cells, Cultured, Epithelium drug effects, Epithelium metabolism, Epithelium pathology, Humans, Interleukin-8 biosynthesis, Interleukin-8 genetics, Male, Oleanolic Acid pharmacology, Prostate drug effects, Prostatic Hyperplasia drug therapy, Prostatic Hyperplasia genetics, RNA, Messenger genetics, RNA, Messenger metabolism, Receptors, CXCR biosynthesis, Receptors, CXCR genetics, Signal Transduction drug effects, Triterpenes pharmacology, Ursolic Acid, Interleukin-8 metabolism, Prostate metabolism, Prostate pathology, Prostatic Hyperplasia metabolism, Prostatic Hyperplasia pathology, Receptors, CXCR metabolism
- Abstract
Background: The clinical manifestation of benign prostatic hyperplasia (BPH) is causally linked to the inflammatory microenvironment and proliferation of epithelial and stromal cells in the prostate transitional zone. The CXC-chemokine interleukin-8 (IL-8) contributes to inflammation. We evaluated the expression of inflammatory cytokines in clinical specimens, primary cultures, and prostatic lineage cell lines. We investigated whether IL-8 via its receptor system (IL-8 axis) promotes BPH., Methods: The messenger RNA and protein expression of chemokines, including components of the IL-8 axis, were measured in normal prostate (NP; n = 7) and BPH (n = 21), urine (n = 24) specimens, primary cultures, prostatic lineage epithelial cell lines (NHPrE1, BHPrE1, BPH-1), and normal prostate cells (RWPE-1). The functional role of the IL-8 axis in prostate epithelial cell growth was evaluated by CRISPR/Cas9 gene editing. The effect of a combination with two natural compounds, oleanolic acid (OA) and ursolic acid (UA), was evaluated on the expression of the IL-8 axis and epithelial cell growth., Results: Among the 19 inflammatory chemokines and chemokine receptors we analyzed, levels of IL-8 and its receptors (CXCR1, CXCR2), as well as, of CXCR7, a receptor for CXCL12, were 5- to 25-fold elevated in BPH tissues when compared to NP tissues (P ≤ .001). Urinary IL-8 levels were threefold to sixfold elevated in BPH patients, but not in asymptomatic males and females with lower urinary tract symptoms (P ≤ .004). The expression of the IL-8 axis components was confined to the prostate luminal epithelial cells in both normal and BPH tissues. However, these components were elevated in BPH-1 and primary explant cultures as compared to RWPE-1, NHPrE1, and BHPrE1 cells. Knockout of CXCR7 reduced IL-8, and CXCR1 expression by 4- to 10-fold and caused greater than or equal to 50% growth inhibition in BPH-1 cells. Low-dose OA + UA combination synergistically inhibited the growth of BPH-1 and BPH primary cultures. In the combination, the drug reduction indices for UA and OA were 16.4 and 7852, respectively, demonstrating that the combination was effective in inhibiting BPH-1 growth at significantly reduced doses of UA or OA alone., Conclusion: The IL-8 axis is a promotor of BPH pathogenesis. Low-dose OA + UA combination inhibits BPH cell growth by inducing autophagy and reducing IL-8 axis expression in BPH-epithelial cells., (© 2020 Wiley Periodicals LLC.)
- Published
- 2020
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26. Identifying Sialylation Linkages at the Glycopeptide Level by Glycosyltransferase Labeling Assisted Mass Spectrometry (GLAMS).
- Author
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Zhu H, Wang S, Liu D, Ding L, Chen C, Liu Y, Wu Z, Bollag R, Liu K, Alexander WM, Yin J, Ma C, Li L, and Wang PG
- Subjects
- Amino Acid Sequence, Animals, Azides chemistry, Azides metabolism, Blood Proteins chemistry, Blood Proteins metabolism, Campylobacter jejuni enzymology, Carbohydrate Sequence, Cattle, Chromatography, High Pressure Liquid, Fetuins chemistry, Fetuins metabolism, Glycosylation, Hexosamines chemistry, Hexosamines metabolism, Humans, Isomerism, Sialoglycoproteins metabolism, Bacterial Proteins metabolism, Monomeric GTP-Binding Proteins metabolism, Sialoglycoproteins analysis, Tandem Mass Spectrometry methods
- Abstract
Precise assignment of sialylation linkages at the glycopeptide level is of importance in bottom-up glycoproteomics and an indispensable step to understand the function of glycoproteins in pathogen-host interactions and cancer progression. Even though some efforts have been dedicated to the discrimination of α2,3/α2,6-sialylated isomers, unambiguous identification of sialoglycopeptide isomers is still needed. Herein, we developed an innovative glycosyltransferase labeling assisted mass spectrometry (GLAMS) strategy. After specific enzymatic labeling, oxonium ions from higher-energy C-trap dissociation (HCD) fragmentation of α2,3-sailoglycopeptides then generate unique reporters to distinctly differentiate those of α2,6-sailoglycopeptide isomers. With this strategy, a total of 1236 linkage-specific sialoglycopeptides were successfully identified from 161 glycoproteins in human serum.
- Published
- 2020
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27. Genomic analysis of racial differences in triple negative breast cancer.
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Chang CS, Kitamura E, Johnson J, Bollag R, and Hawthorn L
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- Adult, Female, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Humans, Middle Aged, Neoplasm Grading, Neoplasm Staging, Phenotype, Triple Negative Breast Neoplasms classification, Triple Negative Breast Neoplasms pathology, Black or African American genetics, Biomarkers, Tumor genetics, Genomics methods, Mutation, Triple Negative Breast Neoplasms ethnology, Triple Negative Breast Neoplasms genetics, White People genetics
- Abstract
Triple negative breast cancer (TNBC) is more prevalent in African Americans (AAs), has a more aggressive clinical course including a higher mortality rate and an increased occurrence of metastases. This study was designed to determine if racial differences at the molecular level might explain the more aggressive phenotype in AAs. Mutation profiling, was performed on 51 AA and 77 CA tumor/ normal pairs. Transcript expression analysis was performed on 35AA and 37CA. Genes with high frequency mutation rates such as MUC4 and TP53 were common to both racial populations, however genes that were less frequently mutated differed between the races suggesting that those cause the more aggressive nature of TNBC in AA women. JAK-Stat and HER2 signaling were unique to the AA and PTEN and mTOR were unique to the CA profiles. Many pathways identified by the mutational profiles were predicted to be down-regulated by the transcript expression profiles., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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28. DNA methylation patterns in bladder tumors of African American patients point to distinct alterations in xenobiotic metabolism.
- Author
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Vantaku V, Amara CS, Piyarathna DWB, Donepudi SR, Ambati CR, Putluri V, Tang W, Rajapakshe K, Estecio MR, Terris MK, Castro PD, Ittmann MM, Williams SB, Lerner SP, Sreekumar A, Bollag R, Coarfa C, Kornberg MD, Lotan Y, Ambs S, and Putluri N
- Subjects
- Black or African American genetics, Chromatography, Liquid, Epigenesis, Genetic, Gene Expression Regulation, Neoplastic, Glucuronic Acid analysis, Glucuronic Acid metabolism, Humans, Metabolomics, Promoter Regions, Genetic, S-Adenosylhomocysteine analysis, S-Adenosylhomocysteine metabolism, S-Adenosylmethionine analysis, S-Adenosylmethionine metabolism, Tandem Mass Spectrometry, Urinary Bladder Neoplasms metabolism, White People genetics, CpG Islands, DNA Methylation, Inactivation, Metabolic genetics, Urinary Bladder Neoplasms genetics
- Abstract
Racial/ethnic disparities have a significant impact on bladder cancer outcomes with African American patients demonstrating inferior survival over European-American patients. We hypothesized that epigenetic difference in methylation of tumor DNA is an underlying cause of this survival health disparity. We analyzed bladder tumors from African American and European-American patients using reduced representation bisulfite sequencing (RRBS) to annotate differentially methylated DNA regions. Liquid chromatography-mass spectrometry (LC-MS/MS) based metabolomics and flux studies were performed to examine metabolic pathways that showed significant association to the discovered DNA methylation patterns. RRBS analysis showed frequent hypermethylated CpG islands in African American patients. Further analysis showed that these hypermethylated CpG islands in patients are commonly located in the promoter regions of xenobiotic enzymes that are involved in bladder cancer progression. On follow-up, LC-MS/MS revealed accumulation of glucuronic acid, S-adenosylhomocysteine, and a decrease in S-adenosylmethionine, corroborating findings from the RRBS and mRNA expression analysis indicating increased glucuronidation and methylation capacities in African American patients. Flux analysis experiments with 13C-labeled glucose in cultured African American bladder cancer cells confirmed these findings. Collectively, our studies revealed robust differences in methylation-related metabolism and expression of enzymes regulating xenobiotic metabolism in African American patients indicate that race/ethnic differences in tumor biology may exist in bladder cancer., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2019
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29. Challenges in Interpreting Multiple Monoclonal Bands on Serum Protein Electrophoresis and Serum Immunofixation Electrophoresis: An Illustrative Case Report.
- Author
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Larsen R, Allen S, Thompson TZ, Bollag R, and Singh G
- Subjects
- Aged, Antibodies, Monoclonal analysis, Antibodies, Monoclonal blood, Biomarkers, Blood Viscosity, Female, Humans, Waldenstrom Macroglobulinemia blood, Waldenstrom Macroglobulinemia diagnosis, Blood Proteins analysis, Electrophoresis methods, Immunoelectrophoresis methods
- Published
- 2019
- Full Text
- View/download PDF
30. The co-chaperone UNC45A is essential for the expression of mitotic kinase NEK7 and tumorigenesis.
- Author
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Eisa NH, Jilani Y, Kainth K, Redd P, Lu S, Bougrine O, Abdul Sater H, Patwardhan CA, Shull A, Shi H, Liu K, Elsherbiny NM, Eissa LA, El-Shishtawy MM, Horuzsko A, Bollag R, Maihle N, Roig J, Korkaya H, Cowell JK, and Chadli A
- Subjects
- Breast Neoplasms genetics, Breast Neoplasms pathology, Carcinogenesis genetics, Carcinogenesis pathology, Female, HeLa Cells, Humans, Intracellular Signaling Peptides and Proteins genetics, MCF-7 Cells, Mitosis genetics, NIMA-Related Kinases genetics, Neoplasm Metastasis, Neoplasm Proteins genetics, PC-3 Cells, Breast Neoplasms metabolism, Carcinogenesis metabolism, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Neoplastic, Intracellular Signaling Peptides and Proteins metabolism, NIMA-Related Kinases biosynthesis, Neoplasm Proteins metabolism
- Abstract
Cumulative evidence suggests that the heat shock protein 90 (Hsp90) co-chaperone UNC-45 myosin chaperone A (UNC45A) contributes to tumorigenesis and that its expression in cancer cells correlates with proliferation and metastasis of solid tumors. However, the molecular mechanism by which UNC45A regulates cancer cell proliferation remains largely unknown. Here, using siRNA-mediated gene silencing and various human cells, we report that UNC45A is essential for breast cancer cell growth, but is dispensable for normal cell proliferation. Immunofluorescence microscopy, along with gene microarray and RT-quantitative PCR analyses, revealed that UNC45A localizes to the cancer cell nucleus, where it up-regulates the transcriptional activity of the glucocorticoid receptor and thereby promotes expression of the mitotic kinase NIMA-related kinase 7 (NEK7). We observed that UNC45A-deficient cancer cells exhibit extensive pericentrosomal material disorganization, as well as defects in centrosomal separation and mitotic chromosome alignment. Consequently, these cells stalled in metaphase and cytokinesis and ultimately underwent mitotic catastrophe, phenotypes that were rescued by heterologous NEK7 expression. Our results identify a key role for the co-chaperone UNC45A in cell proliferation and provide insight into the regulatory mechanism. We propose that UNC45A represents a promising new therapeutic target to inhibit cancer cell growth in solid tumor types., (© 2019 Eisa et al.)
- Published
- 2019
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31. Large-scale profiling of serum metabolites in African American and European American patients with bladder cancer reveals metabolic pathways associated with patient survival.
- Author
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Vantaku V, Donepudi SR, Piyarathna DWB, Amara CS, Ambati CR, Tang W, Putluri V, Chandrashekar DS, Varambally S, Terris MK, Davies K, Ambs S, Bollag R, Apolo AB, Sreekumar A, and Putluri N
- Subjects
- Amino Acids blood, Case-Control Studies, Chromatography, Liquid, Female, Humans, Lipids blood, Male, Mass Spectrometry, Metabolic Networks and Pathways, Survival Analysis, Urinary Bladder Neoplasms ethnology, Urinary Bladder Neoplasms mortality, Black or African American statistics & numerical data, Metabolomics methods, Urinary Bladder Neoplasms blood, White People statistics & numerical data
- Abstract
Background: African Americans (AAs) experience a disproportionally high rate of bladder cancer (BLCA) deaths even though their incidence rates are lower than those of other patient groups. Using a metabolomics approach, this study investigated how AA BLCA may differ molecularly from European Americans (EAs) BLCA, and it examined serum samples from patients with BLCA with the aim of identifying druggable metabolic pathways in AA patients., Methods: Targeted metabolomics was applied to measure more than 300 metabolites in serum samples from 2 independent cohorts of EA and AA patients with BLCA and healthy EA and AA controls via liquid chromatography-mass spectrometry, and this was followed by the identification of altered metabolic pathways with a focus on AA BLCA. A subset of the differential metabolites was validated via absolute quantification with the Biocrates AbsoluteIDQ p180 kit. The clinical significance of the findings was further examined in The Cancer Genomic Atlas BLCA data set., Results: Fifty-three metabolites, mainly related to amino acid, lipid, and nucleotide metabolism, were identified that showed significant differences in abundance between AA and EA BLCA. For example, the levels of taurine, glutamine, glutamate, aspartate, and serine were elevated in serum samples from AA patients versus EA patients. By mapping these metabolites to genes, this study identified significant relations with regulators of metabolism such as malic enzyme 3, prolyl 3-hydroxylase 2, and lysine demethylase 2A that predicted patient survival exclusively in AA patients with BLCA., Conclusions: This metabolic profile of serum samples might be used to assess risk progression in AA BLCA. These first-in-field findings describe metabolic alterations in AA BLCA and emphasize a potential biological basis for BLCA health disparities., (© 2019 American Cancer Society.)
- Published
- 2019
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32. Comparing leukapheresis protocols for an AML patient with symptomatic leukostasis.
- Author
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Cline A, Jajosky R, Shikle J, and Bollag R
- Subjects
- Aged, Clinical Protocols standards, Female, Granulocytes cytology, Humans, Leukocyte Count, Leukocytes, Mononuclear cytology, Leukapheresis methods, Leukemia, Myeloid, Acute therapy, Leukostasis therapy
- Abstract
Background: Acute myeloid leukemia (AML) is a malignancy characterized by rapid clonal proliferation of myeloid precursors, which can result in hyperleukocytosis. Leukapheresis can be used to rapidly reduce the white blood cell count (WBC). However, the only FDA cleared device for WBC depletion, the COBE Spectra, will no longer be supported by the manufacturer in 2017, and there are few studies comparing different methods of leukapheresis., Case Report: A 68-year-old African American female was admitted to the hospital for relapse of her AML. Laboratory data demonstrated a WBC count of 291 600/μL and flow cytometry of the peripheral blood demonstrated 85% myeloid blasts. Leukapheresis was ordered to help treat the leukostasis., Methods: Three different apheresis protocols were used to achieve cytoreduction: Spectra Optia mononuclear collection (MNC) protocol, Spectra Optia granulocyte collection (PMN) protocol, and Therakos CELLEX buffy coat collection without return. Due to different inlet flow rates, the procedures were evaluated based on the number of WBCs collected and volume of blood processed (VBP)., Results: The Spectra Optia PMN collected the most WBCs and collected nearly as many WBCs per VBP as the Therakos CELLEX, which had the highest value., Conclusion: To our knowledge, we are reporting the first use of Therakos CELLEX and Spectra Optia PMN protocol for WBC depletion. While the Spectra Optia granulocyte protocol showed the best performance for this AML patient, further studies will be needed to compare the Spectra Optia PMN protocol to the MNC protocol for AML patients., (© 2017 Wiley Periodicals, Inc.)
- Published
- 2018
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33. Fibroblastic reticular cells of the lymphoid tissues modulate T cell activation threshold during homeostasis via hyperactive cyclooxygenase-2/prostaglandin E 2 axis.
- Author
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Yu M, Guo G, Zhang X, Li L, Yang W, Bollag R, and Cui Y
- Subjects
- Animals, Cells, Cultured, Homeostasis, Humans, Mice, Mice, Inbred C57BL, Palatine Tonsil immunology, T-Lymphocytes immunology, Cyclooxygenase 2 metabolism, Dinoprostone metabolism, Lymphocyte Activation, Palatine Tonsil cytology
- Abstract
Fibroblastic reticular cells (FRCs) in the T cell zone of lymph nodes (LNs) are pivotal for T cell survival, mobility, and peripheral tolerance. Here, we demonstrate that during homeostasis, FRCs also suppress T cell activation via producing high level of prostaglandin E
2 (PGE2 ) due to their thousands-fold higher cyclooxygenase-2 (COX-2) expression than immune cells. This hyperactive COX-2/PGE2 -induced suppression is evident during antigen-specific and non-antigen-specific activations. It is implicated as suppressed TCR-signaling cascades, reduced alterations in activation markers, and inhibited cytokine production of freshly isolated T cells or T cells co-cultured with FRCs compared with those cultured without FRCs. Different from T cell dysfunction, this FRC-mediated suppression is surmountable by enhancing the strength of stimulation and is reversible by COX-2 inhibitors. Furthermore, T cells in the FRC environment where Cox-2 is genetic inactivated are more sensitive and rapidly activated upon stimulations than those in WT environment. Significantly, FRCs of human lymphoid organs manifest similar COX-2/PGE2 hyperactivity and T cell suppression. Together, this study identifies a previously unappreciated intrinsic mechanism of FRCs shared between mice and humans for suppressing T cell sensitivity to activation via PGE2, underscoring the importance of FRCs in shaping the suppressive milieu of lymphoid organs during homeostasis.- Published
- 2017
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34. Characterization of rare mammary tumours appearing on the neck of RIII/Sa mice infected with mouse mammary tumour virus.
- Author
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Sarkar NH, Borke JL, and Bollag RJ
- Subjects
- Animals, Blotting, Southern, Mammary Neoplasms, Animal genetics, Mammary Neoplasms, Animal pathology, Mammary Neoplasms, Animal virology, Mice, Microscopy, Electron, Transmission, Mutation, Neck pathology, Oncogenes, Retroviridae Infections complications, Retroviridae Infections pathology, Soft Tissue Neoplasms genetics, Mammary Tumor Virus, Mouse, Soft Tissue Neoplasms pathology, Soft Tissue Neoplasms virology, Tumor Virus Infections complications, Tumor Virus Infections pathology
- Abstract
RIII/Sa and C3H mice harbour milk-borne mouse mammary tumour virus (MMTV) and develop mammary tumours at a high incidence. These mammary tumours usually arise ventrally and/or on the sides of the animals. In the present study, some mice of both strains were observed to have tumours in the dorsal neck area. Histological analysis of the tumours indicated their similarity to mammary tumours induced by MMTV oncogenesis. The neck tumours were found by thin-section electron microscopy to contain both type A and type B particles that are hallmarks of MMTV infection. In addition, the neck tumour DNA possessed insertion mutations of Wnt-1 and Fgf-3 proto-oncogenes, the activation of which play important roles in the development of mouse mammary tumours. These neck tumours appear to be mammary tumours that arise in the context of in-situ mammary tissue, similar to rare 'ectopic' human breast cancers that arise in the axillary region and other sites remote from the breast., (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Published
- 2013
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35. Fine needle aspiration cytology of an endotracheal mass: report of a case with an unusual presentation of anaplastic large cell lymphoma.
- Author
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Bollag R, Ramalingam P, Davis B, and Reid-Nicholson M
- Subjects
- Adult, Biomarkers, Tumor metabolism, Biopsy, Fine-Needle, Humans, Lymph Nodes pathology, Lymphatic Diseases metabolism, Lymphatic Diseases pathology, Lymphoma, Large-Cell, Anaplastic metabolism, Male, Tomography, X-Ray Computed, Tracheal Neoplasms diagnostic imaging, Tracheal Neoplasms metabolism, Lymphoma, Large-Cell, Anaplastic pathology, Tracheal Neoplasms pathology
- Abstract
Background: Anaplastic large cell lymphoma (ALCL) is an uncommon hematolymphoid neoplasm characterized by malignant lymphocytes of T-cell phenotype. It usually affects young patients and may involve a variety of tissues and organs. We herein describe a case of ALCL that presented as an endotracheal mass with associated hilar lymphadenopathy. To the best of our knowledge this is the first case in the literature arising in the trachea. In this location the diagnosis of ALCL can be especially difficult as its pleomorphic cytomorphology mimics that of a carcinoma, which is a more typical neoplasm arising in the trachea., Case: A 26-year-old male presented with hemoptysis and paroxysmal chest pain. Imaging revealed an endotracheal mass and multiple lytic bone lesions. Fine needle aspiration biopsy of the endotracheal mass revealed discohesive malignant cells with abundant, pale cytoplasm and cerebriform, donut-shaped and horseshoe-shaped nuclei. Immunohistochemical studies confirmed the diagnosis of ALCL., Conclusion: ALCL may have an unusual presentation and involve diverse sites, including the trachea. While its cytologic features are straightforward, a high index of suspicion is necessary to ensure accurate diagnosis when it presents in unusual locations.
- Published
- 2010
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36. Prognostic implications of survivin expression in squamous cell carcinoma of the larynx.
- Author
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Vaught C, Weinberger P, Seybt TP, Bollag R, and Jackson L
- Subjects
- Biomarkers, Tumor blood, Enzyme-Linked Immunosorbent Assay, Female, Humans, Immunohistochemistry, Inhibitor of Apoptosis Proteins, Male, Middle Aged, Prognosis, Retrospective Studies, Survivin, Carcinoma, Squamous Cell metabolism, Laryngeal Neoplasms metabolism, Microtubule-Associated Proteins blood
- Published
- 2010
- Full Text
- View/download PDF
37. Metastatic hepatocellular carcinoma with CD138 positivity: an unusual mimic of multiple myeloma?
- Author
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Ramalingam P, Adeagbo B, Bollag R, Lee J, and Reid-Nicholson M
- Subjects
- Aged, Biopsy, Fine-Needle, Carcinoma, Hepatocellular secondary, Diagnosis, Differential, Gene Expression Regulation, Neoplastic, Humans, Liver Neoplasms secondary, Male, Syndecan-1 genetics, Carcinoma, Hepatocellular diagnosis, Carcinoma, Hepatocellular immunology, Liver Neoplasms diagnosis, Liver Neoplasms immunology, Multiple Myeloma diagnosis, Syndecan-1 metabolism
- Abstract
CD138 is a monoclonal anti-syndecan-1 antibody that is often used to identify plasma cells in the bone marrow of patients with multiple myeloma (MM). Several carcinomas may also express CD138 including prostate, colon, renal cell, and hepatocellular carcinoma (HCC). We report a case of metastatic HCC that presented as a soft tissue mass on the back of a 67-year-old male. Based on the clinical and radiologic findings, MM was strongly suspected. In addition, fine-needle aspiration biopsy (FNAB) of the mass revealed neoplastic cells that were positive for CD138, both by immunohistochemistry (IHC) and flow cytometry. The cytomorphologic features however did not support a diagnosis of MM, but were consistent with metastatic HCC. Our case highlights the potential problems that may arise by over-reliance on IHC and flow cytometry. Careful morphologic assessment as well as clinical and radiologic correlation are very important when evaluating any CD138-positive neoplasm. This approach should improve diagnostic accuracy and reduce the risk of erroneous interpretation of aberrant IHC results. In addition, we examined the expression of CD138 in known cases of HCC.
- Published
- 2008
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38. The solitary long terminal repeats of ERV-9 endogenous retrovirus are conserved during primate evolution and possess enhancer activities in embryonic and hematopoietic cells.
- Author
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Ling J, Pi W, Bollag R, Zeng S, Keskintepe M, Saliman H, Krantz S, Whitney B, and Tuan D
- Subjects
- Animals, Base Sequence, Cell Line, Embryo, Mammalian cytology, Flow Cytometry, Genes, Reporter, Globins genetics, Hematopoietic Stem Cells cytology, Humans, Introns genetics, Molecular Sequence Data, Promoter Regions, Genetic genetics, Retroelements genetics, Transfection, Biological Evolution, Conserved Sequence, Endogenous Retroviruses genetics, Enhancer Elements, Genetic genetics, Primates genetics, Terminal Repeat Sequences genetics
- Abstract
The solitary long terminal repeats (LTRs) of ERV-9 endogenous retrovirus contain the U3, R, and U5 regions but no internal viral genes. They are middle repetitive DNAs present at 2,000 to 4,000 copies in primate genomes. Sequence analyses of the 5" boundary area of the erythroid beta-globin locus control region (beta-LCR) and the intron of the embryonic axin gene show that a solitary ERV-9 LTR has been stably integrated in the respective loci for at least 15 million years in the higher primates from orangutan to human. Functional studies utilizing the green fluorescent protein (GFP) gene as the reporter in transfection experiments show that the U3 region of the LTRs possesses strong enhancer activity in embryonic cells of widely different tissue origins and in adult cells of blood lineages. In both the genomic LTRs of embryonic placental cells and erythroid K562 cells and transfected LTRs of recombinant GFP plasmids in K562 cells, the U3 enhancer activates synthesis of RNAs that are initiated from a specific site 25 bases downstream of the AATAAA (TATA) motif in the U3 promoter. A second AATAAA motif in the R region does not serve as the TATA box or as the polyadenylation signal. The LTR-initiated RNAs extend through the R and U5 regions into the downstream genomic DNA. The results suggest that the ERV-9 LTR-initiated transcription process may modulate transcription of the associated gene loci in embryonic and hematopoietic cells.
- Published
- 2002
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39. 1,25-Dihydroxyvitamin D(3), phospholipase D and protein kinase C in keratinocyte differentiation.
- Author
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Bollinger Bollag W and Bollag RJ
- Subjects
- Amino Acid Sequence, Animals, Calcitriol pharmacology, Cell Differentiation drug effects, Consensus Sequence, Humans, Molecular Sequence Data, Phospholipase D drug effects, Phospholipase D pharmacology, Protein Kinase C drug effects, Protein Kinase C pharmacology, Signal Transduction drug effects, Skin Diseases etiology, Calcitriol physiology, Keratinocytes cytology, Phospholipase D physiology, Protein Kinase C physiology
- Abstract
1,25-Dihydroxyvitamin D(3), thought to be a physiological regulator of epidermal keratinocyte growth and differentiation, also elicits the complete differentiative program in vitro, with expression of various genes/proteins characteristic of both early and late differentiation. 1,25-Dihydroxyvitamin D(3) functions by interacting with an intracellular receptor that binds to DNA at vitamin D response elements (VDRE) thereby affecting transcription. 1,25-Dihydroxyvitamin D(3) has been demonstrated to alter the expression of several enzymes involved in signal transduction, and presumably this is the mechanism through which the hormone regulates differentiation. It has recently been shown that 1,25-dihydroxyvitamin D(3) specifically increases the expression/activity of phospholipase D-1, an enzyme that hydrolyzes phospholipids to generate lipid messengers, such as diacylglycerol (DAG). DAG, in turn, is known to activate several members of the protein kinase C (PKC) family. It has been proposed that this signaling pathway mediates late differentiation events in epidermal keratinocytes. In this article the data supporting a role for PKC and phospholipase D in keratinocyte differentiation, as well as in the pathogenesis of skin diseases, are reviewed and a model is proposed for the signaling pathways that regulate this process upon exposure to 1,25-dihydroxyvitamin D(3).
- Published
- 2001
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40. Functional parathyroid hormone receptors are present in an umbilical vein endothelial cell line.
- Author
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Isales CM, Sumpio B, Bollag RJ, Zhong Q, Ding KH, Du W, Rodriguez-Commes J, Lopez R, Rosales OR, Gasalla-Herraiz J, McCarthy R, and Barrett PQ
- Subjects
- Calcium metabolism, Cell Line, Chromatography, High Pressure Liquid, Cyclic AMP biosynthesis, Endothelin-1 metabolism, Endothelium, Vascular cytology, Fluorescent Dyes, Fura-2, Glyceraldehyde-3-Phosphate Dehydrogenases metabolism, Humans, Inositol Phosphates metabolism, Parathyroid Hormone metabolism, RNA, Messenger biosynthesis, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction physiology, Thymidine metabolism, Umbilical Veins cytology, Endothelium, Vascular metabolism, Receptors, Parathyroid Hormone metabolism, Umbilical Veins metabolism
- Abstract
Acute parathyroid hormone exposure induces vascular smooth muscle relaxation. In contrast, continuous infusion of parathyroid hormone leads to vasoconstriction and an elevation of blood pressure. Despite the known effects of parathyroid hormone on vascular smooth muscle, possible direct effects on the vascular endothelium have not previously been investigated. Using a human umbilical vein endothelial cell line, we found that parathyroid hormone increased both intracellular calcium and cellular cAMP content in these endothelial cells. Furthermore, exposure of these cells to increasing concentrations of parathyroid hormone stimulated both [(3)H]thymidine incorporation and endothelin-1 secretion. Parathyroid hormone/parathyroid hormone-related peptide receptor mRNA could be detected at low levels in these cells. In summary, these data demonstrate that endothelium-derived cells contain functional parathyroid hormone receptors. The potential physiological role of these receptors remains to be determined.
- Published
- 2000
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41. Use of a repetitive mouse B2 element to identify transplanted mouse cells in mouse-chick chimeras.
- Author
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Bollag RJ, Crawford KB, Stadt H, Kumiski D, Zdanowicz M, Baptista C, Herlea V, and Kirby ML
- Subjects
- Animals, Cell Transplantation, Chick Embryo, Mice, Cell Movement physiology, Repetitive Sequences, Nucleic Acid
- Abstract
Monitoring the migrations of cells during embryonic development requires a system in which cells can be identified in situ during locomotion. One promising system involves the generation of chimeras by transplanting mouse cells into chick embryos in ovo to exploit the wealth of mouse genetic variants. The success of this technique relies on the ability to detect individual mouse cells in a chick environment with high specificity. The murine B2 family of short interspersed elements is present in the mouse genome at copy numbers in excess of 10(5), whereas this sequence is absent in the chick genome based on hybridization techniques. This differential of five orders of magnitude produces signals in mouse cells that are easily identified, even in an environment that is predominantly chick. Thus, the B2 repeat probe is highly effective for the purpose of identifying mouse cells in mouse-chick chimeras., (Copyright 1999 Academic Press.)
- Published
- 1999
- Full Text
- View/download PDF
42. Identification of caveolin and caveolin-related proteins in the brain.
- Author
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Cameron PL, Ruffin JW, Bollag R, Rasmussen H, and Cameron RS
- Subjects
- Animals, Antibodies, Monoclonal, Antibody Affinity, Astrocytes ultrastructure, Biological Transport physiology, Blotting, Northern, Brain cytology, Caveolin 1, Cell Compartmentation physiology, Cell Fractionation, Cell Membrane chemistry, Cell Membrane metabolism, Detergents, Female, Fluorescent Antibody Technique, Indirect, Gene Expression Regulation, Developmental physiology, Intracellular Membranes chemistry, Intracellular Membranes metabolism, Membrane Proteins genetics, Membrane Proteins immunology, Microscopy, Electron, Microtomy, Nerve Tissue Proteins analysis, Nerve Tissue Proteins genetics, Nerve Tissue Proteins immunology, Oligodendroglia ultrastructure, Pregnancy, RNA, Messenger analysis, Rats, Rats, Sprague-Dawley, Solubility, Astrocytes chemistry, Caveolins, Cell Membrane ultrastructure, Membrane Proteins analysis, Oligodendroglia chemistry
- Abstract
Caveolae are 50-100 nm, nonclathrin-coated, flask-shaped plasma membrane microdomains that have been identified in most mammalian cell types, except lymphocytes and neurons. To date, multiple functions have been ascribed to caveolae, including the compartmentalization of lipid and protein components that function in transmembrane signaling events, biosynthetic transport functions, endocytosis, potocytosis, and transcytosis. Caveolin, a 21-24 kDa integral membrane protein, is the principal structural component of caveolae. We have initiated studies to examine the relationship of detergent-insoluble complexes identified in astrocytes to the caveolin-caveolae compartment detected in cells of peripheral tissues. Immunolocalization studies performed in astrocytes reveal caveolin immunoreactivity in regions that correlate well to the distribution of caveolae and caveolin determined in other cell types, and electron microscopic studies reveal multiple clusters of flask-shaped invaginations aligned along the plasma membrane. Immunoblot analyses demonstrate that detergent-insoluble complexes isolated from astrocytes are composed of caveolin-1alpha, an identification verified by Northern blot analyses and by the cloning of a cDNA using reverse transcriptase-PCR amplification from total astrocyte RNA. Using a full-length caveolin-1 probe, Northern blot analyses suggest that the expression of caveolin-1 may be regulated during brain development. Immunoblot analyses of detergent-insoluble complexes isolated from cerebral cortex and cerebellum identify two immunoreactive polypeptides with apparent molecular weight and isoelectric points appropriate for caveolin. The identification of caveolae microdomains and caveolin-1 in astrocytes and brain, as well as the apparent regulation of caveolin-1 expression during brain development, identifies a cell compartment not detected previously in brain.
- Published
- 1997
43. Cloning and expression analysis of murine phospholipase D1.
- Author
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Colley WC, Altshuller YM, Sue-Ling CK, Copeland NG, Gilbert DJ, Jenkins NA, Branch KD, Tsirka SE, Bollag RJ, Bollag WB, and Frohman MA
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Chromosome Mapping, Cloning, Molecular, Gene Expression, In Situ Hybridization, Mice, Molecular Sequence Data, Organ Specificity, Phospholipase D metabolism, Sequence Alignment, Phospholipase D genetics
- Abstract
Activation of phosphatidylcholine-specific phospholipase D(PLD) occurs as part of the complex signal-transduction cascade initiated by agonist stimulation of tyrosine kinase and G-protein-coupled receptors. A variety of mammalian PLD activities have been described, and cDNAs for two PLDs recently reported (human PLD1 and murine PLD2). We describe here the cloning and chromosomal localization of murine PLD1. Northern-blot hybridization and RNase protection analyses were used to examine the expression of murine PLD1 and PLD2 ina variety of cell lines and tissues. PLD1 and PLD2 were expressed in all RNA samples examined, although the absolute expression of each isoform varied, as well as the ratio of PLD1 to PLD2. Moreover, in situ hybridization of adult brain and murine embryo sections revealed high levels of expression of individual PLDs in some cell types and no detectable expression in others. Thus the two PLDs probably carry out distinct roles in restricted subsets of cells rather than ubiquitous roles in all cells.
- Published
- 1997
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44. Evolution of mouse T-box genes by tandem duplication and cluster dispersion.
- Author
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Agulnik SI, Garvey N, Hancock S, Ruvinsky I, Chapman DL, Agulnik I, Bollag R, Papaioannou V, and Silver LM
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Conserved Sequence, DNA, Complementary, Evolution, Molecular, Mice, Molecular Sequence Data, Phylogeny, Sequence Homology, Amino Acid, DNA-Binding Proteins genetics, Multigene Family, Repetitive Sequences, Nucleic Acid, T-Box Domain Proteins, Transcription Factors genetics
- Abstract
The T-box genes comprise an ancient family of putative transcription factors conserved across species as divergent as Mus musculus and Caenorhabditis elegans. All T-box gene products are characterized by a novel 174-186-amino acid DNA binding domain called the T-box that was first discovered in the polypeptide products of the mouse T locus and the Drosophila melanogaster optomotor-blind gene. Earlier studies allowed the identification of five mouse T-box genes, T, Tbx1-3, and Tbr1, that all map to different chromosomal locations and are expressed in unique temporal and spatial patterns during embryogenesis. Here, we report the discovery of three new members of the mouse T-box gene family, named Tbx4, Tbx5, and Tbx6. Two of these newly discovered genes, Tbx4 and Tbx5, were found to be tightly linked to previously identified T-box genes. Combined results from phylogenetic, linkage, and physical mapping studies provide a picture for the evolution of a T-box subfamily by unequal crossing over to form a two-gene cluster that was duplicated and dispersed to two chromosomal locations. This analysis suggests that Tbx4 and Tbx5 are cognate genes that diverged apart from a common ancestral gene during early vertebrate evolution.
- Published
- 1996
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45. Expression of the T-box family genes, Tbx1-Tbx5, during early mouse development.
- Author
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Chapman DL, Garvey N, Hancock S, Alexiou M, Agulnik SI, Gibson-Brown JJ, Cebra-Thomas J, Bollag RJ, Silver LM, and Papaioannou VE
- Subjects
- Animals, In Situ Hybridization, Mice, Mice, Mutant Strains, DNA-Binding Proteins genetics, Embryonic and Fetal Development genetics, Gene Expression Regulation, Developmental, T-Box Domain Proteins
- Abstract
A novel family of genes, characterized by the presence of a region of homology to the DNA-binding domain of the Brachyury (T) locus product, has recently been identified. The region of homology has been named the T-box, and the new mouse genes that contain the T-box domain have been named T-box 1-6 (Tbx1 through Tbx6). As the basis for further study of the function and evolution of these genes, we have examined the expression of 5 of these genes, Tbx1-Tbx5, across a wide range of embryonic stages from blastocyst through gastrulation and early organogenesis by in situ hybridization of wholemounts and tissue sections. Tbx3 is expressed earliest, in the inner cell mass of the blastocyst. Four of the genes are expressed in different components of the mesoderm or mesoderm/endoderm during gastrulation (Tbx1 and Tbx3-5). All of these genes have highly specific patterns of expression during later embryogenesis, notably in areas undergoing inductive tissue interactions. In several cases there is complementary expression of different genes in 2 interacting tissues, as in the lung epithelium (Tbx1) and lung mesenchyme (Tbx2-5), and in mammary buds (Tbx3) and mammary stroma (Tbx2). Tbx1 shows very little overlap in the sites of expression with the other 4 genes, in contrast to a striking similarity in expression between members of the 2 cognate gene sets, Tbx2/Tbx3 and Tbx4/Tbx5. This is a clear reflection of the evolutionary relationship between the 5 genes since the divergence of Tbx1 occurred long before the relatively recent divergence of Tbx2 and 3 and Tbx4 and 5 from common ancestral genes. These studies are a good indication that the T-box family of genes has important roles in inductive interactions in many stages of mammalian embryogenesis.
- Published
- 1996
- Full Text
- View/download PDF
46. Conservation of the T-box gene family from Mus musculus to Caenorhabditis elegans.
- Author
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Agulnik SI, Bollag RJ, and Silver LM
- Subjects
- Amino Acid Sequence, Animals, Cloning, Molecular, DNA, Complementary, Databases, Factual, Drosophila melanogaster genetics, Gene Library, Molecular Sequence Data, Phylogeny, Sequence Homology, Amino Acid, Species Specificity, Xenopus laevis genetics, Zebrafish genetics, Biological Evolution, Caenorhabditis elegans genetics, Conserved Sequence, Mice genetics, Multigene Family
- Abstract
Recently, a novel family of genes with a region of homology to the mouse T locus, which is known to play a crucial, and conserved, role in vertebrate development, has been discovered. The region of homology has been named the T-box. The T-box domain of the prototypical T locus product is associated with sequence-specific DNA binding activity. In this report, we have characterized four members of the T-box gene family from the nematode Caenorhabditis elegans. All lie in close proximity to each other in the middle of chromosome III. Homology analysis among all completely sequenced T-box products indicates a larger size for the conserved T-box domain (166 to 203 residues) than previously reported. Phylogenetic analysis suggests that one C. elegans T-box gene may be a direct ortholog of the mouse Tbx2 and Drosophila omb genes. The accumulated data demonstrate the ancient nature of the T-box gene family and suggest the existence of at least three separate T-box-containing genes in a common early metazoan ancestor to nematodes and vertebrates.
- Published
- 1995
- Full Text
- View/download PDF
47. An ancient family of embryonically expressed mouse genes sharing a conserved protein motif with the T locus.
- Author
-
Bollag RJ, Siegfried Z, Cebra-Thomas JA, Garvey N, Davison EM, and Silver LM
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins chemistry, DNA-Binding Proteins physiology, Drosophila genetics, Fetal Proteins biosynthesis, Fetal Proteins chemistry, Fetal Proteins physiology, Gene Expression Regulation, Genes, Genes, Insect, Genes, Lethal, Invertebrates embryology, Invertebrates growth & development, Mesoderm, Mice embryology, Mice growth & development, Molecular Sequence Data, Nerve Tissue Proteins genetics, Phylogeny, Polymerase Chain Reaction, Sequence Alignment, Sequence Homology, Amino Acid, Species Specificity, Vertebrates embryology, Vertebrates growth & development, DNA-Binding Proteins genetics, Drosophila Proteins, Embryonic and Fetal Development genetics, Fetal Proteins genetics, Invertebrates genetics, Mice genetics, Multigene Family, Protein Structure, Tertiary, T-Box Domain Proteins, Vertebrates genetics
- Abstract
The T locus encodes a product with DNA binding activity that is likely to play a role in the development of all vertebrate organisms. We have identified and characterized a novel family of mouse genes that share a protein motif, the T-box, with the prototypical T locus. The T-box domain of the T locus co-localizes with its DNA binding activity. Each T-box gene is expressed in a unique temporal and spatial pattern during embryogenesis. Phylogenetic analysis suggests that at least three T-box genes were present in the common ancestor to vertebrates and invertebrates. Thus, members of the T-box family could have played a role in the evolution of all metazoan organisms.
- Published
- 1994
- Full Text
- View/download PDF
48. Homologous recombination in mammalian cells.
- Author
-
Bollag RJ, Waldman AS, and Liskay RM
- Subjects
- Animals, Chromosome Mapping, DNA genetics, Extrachromosomal Inheritance, Mammals genetics, Recombination, Genetic
- Published
- 1989
- Full Text
- View/download PDF
49. A new skin cleanser, Emavon.
- Author
-
BOLLAG R
- Subjects
- Humans, Dermatologic Agents, Detergents, Skin
- Published
- 1947
50. [Perifolliculitis abscedens and suffodiens].
- Author
-
MIESCHER G and BOLLAG R
- Subjects
- Humans, Dermatitis, Folliculitis
- Published
- 1952
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