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  • 1
    Publication Date: 2008-08-01
    Description: Many advanced snakes use fangs-specialized teeth associated with a venom gland-to introduce venom into prey or attacker. Various front- and rear-fanged groups are recognized, according to whether their fangs are positioned anterior (for example cobras and vipers) or posterior (for example grass snakes) in the upper jaw. A fundamental controversy in snake evolution is whether or not front and rear fangs share the same evolutionary and developmental origin. Resolving this controversy could identify a major evolutionary transition underlying the massive radiation of advanced snakes, and the associated developmental events. Here we examine this issue by visualizing the tooth-forming epithelium in the upper jaw of 96 snake embryos, covering eight species. We use the sonic hedgehog gene as a marker, and three-dimensionally reconstruct the development in 41 of the embryos. We show that front fangs develop from the posterior end of the upper jaw, and are strikingly similar in morphogenesis to rear fangs. This is consistent with their being homologous. In front-fanged snakes, the anterior part of the upper jaw lacks sonic hedgehog expression, and ontogenetic allometry displaces the fang from its posterior developmental origin to its adult front position-consistent with an ancestral posterior position of the front fang. In rear-fanged snakes, the fangs develop from an independent posterior dental lamina and retain their posterior position. In light of our findings, we put forward a new model for the evolution of snake fangs: a posterior subregion of the tooth-forming epithelium became developmentally uncoupled from the remaining dentition, which allowed the posterior teeth to evolve independently and in close association with the venom gland, becoming highly modified in different lineages. This developmental event could have facilitated the massive radiation of advanced snakes in the Cenozoic era, resulting in the spectacular diversity of snakes seen today.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Vonk, Freek J -- Admiraal, Jeroen F -- Jackson, Kate -- Reshef, Ram -- de Bakker, Merijn A G -- Vanderschoot, Kim -- van den Berge, Iris -- van Atten, Marit -- Burgerhout, Erik -- Beck, Andrew -- Mirtschin, Peter J -- Kochva, Elazar -- Witte, Frans -- Fry, Bryan G -- Woods, Anthony E -- Richardson, Michael K -- England -- Nature. 2008 Jul 31;454(7204):630-3. doi: 10.1038/nature07178.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Institute of Biology, Leiden University, Kaiserstraat 63, PO Box 9516, 2300 RA, Leiden, The Netherlands.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/18668106" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Gene Expression Regulation, Developmental ; Hedgehog Proteins/genetics/metabolism ; In Situ Hybridization ; Molecular Sequence Data ; *Phylogeny ; Snakes/anatomy & histology/classification/*embryology/genetics ; Tooth/anatomy & histology/*embryology
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 2
    Publication Date: 2008-07-18
    Description: 〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Vonk, Freek J -- Richardson, Michael K -- England -- Nature. 2008 Jul 17;454(7202):282-3. doi: 10.1038/454282a.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/18633402" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Body Patterning/*physiology ; Mice ; Snakes/*embryology ; Somites/*embryology ; Time Factors
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 3
    Publication Date: 2008-02-15
    Description: Hodgkin and Reed-Sternberg (HRS) cells in Hodgkin lymphoma (HL) secrete factors that interact with inflammatory background cells and may serve as biomarkers for disease activity. To detect new proteins related to pathogenesis, we analyzed the secretome of HRS cells. Proteins in cell culture supernatant of 4 HL cell lines were identified using 1DGE followed by in-gel trypsin digestion and LC-MS/MS. In total, 1290 proteins, including 368 secreted proteins, were identified. Functional grouping of secreted proteins revealed 37 proteins involved in immune response. Sixteen of the 37 proteins (ie, ALCAM, Cathepsin C, Cathepsin S, CD100, CD150, CD26, CD44, CD63, CD71, Fractal-kine, IL1R2, IL25, IP-10, MIF, RANTES, and TARC) were validated in HL cell lines and patient material using immunohistochemistry and/or ELISA. Expression of all 16 proteins was confirmed in HL cell lines, and 15 were also confirmed in HL tissues. Seven proteins (ALCAM, cathepsin S, CD26, CD44, IL1R2, MIF, and TARC) revealed significantly elevated levels in patient plasma compared with healthy controls. Proteomics analyses of HL cell line supernatant allowed detection of new secreted proteins, which may add to our insights in the interaction between HRS cells and infiltrating lymphocytes and in some instances might serve as biomarkers.
    Print ISSN: 0006-4971
    Electronic ISSN: 1528-0020
    Topics: Biology , Medicine
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  • 4
    Publication Date: 2006-11-01
    Description: Hodgkin lymphoma (HL) is characterized by a minority of neoplastic cells, the Hodgkin and Reed-Sternberg cells (HRS cells) and an inflammatory background. Abnormal cytokine and chemokine production may play an important role in the proliferation of HRS cells, the reactive background formation and the impaired immune response encountered in patients with HL. To detect new proteins that might be involved in the interaction between the HRS cells and the inflammatory background cells and that also might serve as useful biomarkers for HL, we used different proteomic techniques to study the proteome of HRS cells. Methods: The first step is to determine the secretome of HRS cells in vitro. Therefore, the cells needed to be cultured without serum. Hodgkin cell lines L428 and L1236 were washed extensively to deplete serum proteins and cultured in RPMI media without serum for 6–72 hours. The supernatant was collected, concentrated, and analyzed by Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-TOF-MS) to determine the protein composition and as a quality check. After culture process optimization, sample fractionation was carried out by SDS-PAGE. The whole lane was excised in 30–35 bands and trypsine digestion was performed on every band. Peptide mixtures were analyzed by LC-MS/MS for identification of the proteins. Results: We tested several culture approaches for the Hodgkin lymphoma cell lines for proteomics purpose. Serum-free media were found to be inappropriate because of the abundant presence of several proteins. We next tested efficiency of washing steps and culture characteristics of 5 Hodgkin lymphoma cell lines in RPMI without serum. Cell viability, TARC and MDC levels in the culture supernatant at different time points and also SELDI-TOF-MS spectra indicated that culturing for 6–24 hours gave the best results. For protein identification we started with analysis of small proteins from supernatant of the L428 cell line, which has the highest TARC production. Gel bands in the region between 6–14 kDa were isolated from SDS-PAGE and treated for protein identification. This revealed a total of 64 proteins identified with ≥95% confidence. Application of SecretomeP2.0 (Protein Eng.Des.Sel., 2004. TMHMM 2.0) software indicated the presence of 28 secreted proteins including TARC. A similar approach was used for the L1236 Hodgkin lymphoma cell line, but the whole mass range (3–188 kDa) was analyzed in this case. We identified a total of 215 different proteins, out of which 85 are secreted (24 proteins follow a classical pathway while the other 61 follow a non-classical pathway). The non-secreted proteins were localized to the ER lumen, mitochondrion, nucleus, and membrane. These proteins are probably derived from cells breaking up during culture or centrifugation. Conclusion: In this study we identified 85 secreted proteins in L1236 supernatant, and 28 secreted proteins in the low molecular mass range (6–14kDa) in L428 supernatant. Some of the secreted proteins, including TARC, macrophage migration inhibitory factor (MIF), CD44 and Activated Leukocyte Cell Adhesion Molecule (ALCAM), are known to be involved in inflammatory process, lymphocytes homing or cell adhesion. The most consistently observed proteins will be tested for potential use as biomarkers in the sera of HL patients.
    Print ISSN: 0006-4971
    Electronic ISSN: 1528-0020
    Topics: Biology , Medicine
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