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Simões, Inês T., Fernando Aranda, Sergi Casadó-Llombart, María Velasco-de Andrés, Cristina Català, Pilar Álvarez, Marta Consuegra-Fernández i in. "Multifaceted effects of soluble human CD6 in experimental cancer models". Journal for ImmunoTherapy of Cancer 8, nr 1 (marzec 2020): e000172. http://dx.doi.org/10.1136/jitc-2019-000172.
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BackgroundCD6 is a lymphocyte surface co-receptor physically associated with the T-cell receptor (TCR)/CD3 complex at the center of the immunological synapse. There, CD6 assists in cell-to-cell contact stabilization and modulation of activation/differentiation events through interaction with CD166/ALCAM (activated leukocyte cell adhesion molecule), its main reported ligand. While accumulating evidence is attracting new interest on targeting CD6 for therapeutic purposes in autoimmune disorders, little is known on its potential in cancer. In an attempt to elucidate the in vivo relevance of blocking CD6-mediated interactions in health and disease, we explored the consequences of expressing high circulating levels of a soluble form CD6 (sCD6) as a decoy receptor.MethodsHigh sCD6 serum levels were achieved by using transgenic C57BL/6 mice expressing human sCD6 under the control of lymphoid-specific transcriptional elements (shCD6LckEμTg) or wild type either transduced with hepatotropic adeno-associated virus coding for mouse sCD6 or undergoing repeated infusions of recombinant human sCD6 protein. Characterization of sCD6-induced changes was performed by ex vivo flow cytometry and functional analyses of mouse lymphoid organ cells. The in vivo relevance of those changes was explored by challenging mice with subcutaneous or metastatic tumors induced by syngeneic cancer cells of different lineage origins.ResultsThrough a combination of in vitro and in vivo studies, we show that circulating sCD6 expression induces defective regulatory T cell (Treg) generation and function, decreased CD166/ALCAM-mediated tumor cell proliferation/migration and impaired galectin-induced T-cell apoptosis, supporting the fact that sCD6 modulates antitumor lymphocyte effector function and tumorigenesis. Accordingly, sCD6 expression in vivo resulted in delayed subcutaneous tumor growth and/or reduced metastasis on challenge of mice with syngeneic cancer cells.ConclusionsEvidence is provided for the disruption of CD6 receptor–ligand interactions as a feasible immunomodulatory approach in cancer.
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Krüger, Timothy, Mario Hofweber i Susanne Kramer. "SCD6 induces ribonucleoprotein granule formation in trypanosomes in a translation-independent manner, regulated by its Lsm and RGG domains". Molecular Biology of the Cell 24, nr 13 (lipiec 2013): 2098–111. http://dx.doi.org/10.1091/mbc.e13-01-0068.
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Ribonucleoprotein (RNP) granules are cytoplasmic, microscopically visible structures composed of RNA and protein with proposed functions in mRNA decay and storage. Trypanosomes have several types of RNP granules, but lack most of the granule core components identified in yeast and humans. The exception is SCD6/Rap55, which is essential for processing body (P-body) formation. In this study, we analyzed the role of trypanosome SCD6 in RNP granule formation. Upon overexpression, the majority of SCD6 aggregates to multiple granules enriched at the nuclear periphery that recruit both P-body and stress granule proteins, as well as mRNAs. Granule protein composition depends on granule distance to the nucleus. In contrast to findings in yeast and humans, granule formation does not correlate with translational repression and can also take place in the nucleus after nuclear targeting of SCD6. While the SCD6 Lsm domain alone is both necessary and sufficient for granule induction, the RGG motif determines granule type and number: the absence of an intact RGG motif results in the formation of fewer granules that resemble P-bodies. The differences in granule number remain after nuclear targeting, indicating translation-independent functions of the RGG domain. We propose that, in trypanosomes, a local increase in SCD6 concentration may be sufficient to induce granules by recruiting mRNA. Proteins that bind selectively to the RGG and/or Lsm domain of SCD6 could be responsible for regulating granule type and number.
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Decker, Carolyn J., i Roy Parker. "CAR-1 and Trailer hitch: driving mRNP granule function at the ER?" Journal of Cell Biology 173, nr 2 (24.04.2006): 159–63. http://dx.doi.org/10.1083/jcb.200601153.
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The targeting of messenger RNAs (mRNAs) to specific subcellular sites for local translation plays an important role in diverse cellular and developmental processes in eukaryotes, including axis formation, cell fate determination, spindle pole regulation, cell motility, and neuronal synaptic plasticity. Recently, a new conserved class of Lsm proteins, the Scd6 family, has been implicated in controlling mRNA function. Depletion or mutation of members of the Scd6 family, Caenorhabditis elegans CAR-1 and Drosophila melanogaster trailer hitch, lead to a variety of developmental phenotypes, which in some cases can be linked to alterations in the endoplasmic reticulum (ER). Scd6/Lsm proteins are RNA binding proteins and are found in RNP complexes associated with translational control of mRNAs, and these complexes can colocalize with the ER. These findings raise the possibility that localization and translational regulation of mRNAs at the ER plays a role in controlling the organization of this organelle.
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Ramos‐Casals, M., J. Font, M. García‐Carrasco, J. Calvo, L. Places, O. Padilla, R. Cervera, M. A. Bowen, F. Lozano i M. Ingelmo. "High circulating levels of soluble scavenger receptors (sCD5 and sCD6) in patients with primary Sjögren's syndrome". Rheumatology 40, nr 9 (wrzesień 2001): 1056–59. http://dx.doi.org/10.1093/rheumatology/40.9.1056.
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Cristodero, Marina, Bernd Schimanski, Manfred Heller i Isabel Roditi. "Functional characterization of the trypanosome translational repressor SCD6". Biochemical Journal 457, nr 1 (10.12.2013): 57–67. http://dx.doi.org/10.1042/bj20130747.
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Trypanosomal SCD6 is a general repressor of translation. It is not required for stress granule formation and, unusually, does not interact with the helicase DHH1. We analysed domains involved in the localization and function of TbSCD6 and identified interacting partners.
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Bhatter, Nupur, Rajan Iyyappan i Purusharth I. Rajyaguru. "Characterizing mutations in and genetic interactions of RGG-motif translation repressor Sbp1". Wellcome Open Research 3 (22.08.2018): 102. http://dx.doi.org/10.12688/wellcomeopenres.14709.1.
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Background: Mechanisms of mRNA fate decisions play an important role in determining if a given mRNA will be translated, stored or degraded upon arrival to cytoplasm. Sbp1 is an important RGG-motif containing protein that is implicated in mRNA fate decisions since it can affect mRNA decapping and translation. Sbp1 represses translation by binding eIF4G1 through its RGG-motif and activates decapping when overexpressed. In order to understand the amino acids important for translation repression activity of Sbp1 we performed mutational analysis of Sbp1 combined with assessing its genetic interaction with another RGG-motif protein Scd6. We created two classes of point mutations a) in aromatic residues of the RGG-motif and b) in residues reported to be phosphorylated. Method: Sequence alignment was performed to identify aromatic residues to be mutated based on conservation. Site-directed mutagenesis approach was used to create several point mutations in Sbp1 expressed under galactose-inducible promoter. The mutants were tested for their ability to cause growth defect upon overexpression. The ability of Sbp1 to affect repression activity of other decapping activators was tested using the same growth assay. Results: Mutation of several aromatic residues in the RGG-motif of Sbp1 led to a weak rescue phenotype. However the phospho-mimetic mutants of Sbp1 did not lead to any kind of growth defect rescue. Deletion of another eIF4G1-binding RGG-motif protein Scd6 does not affect ability of Sbp1 to cause growth defect. On the other hand absence of Sbp1 does not affect ability of Dhh1 and Pat1 to repress translation. Conclusion: Based on our growth assay analysis we conclude that mutated aromatic residues contribute marginally to repression activity of Sbp1 whereas phospho-mimetic mutants do not alter ability of Sbp1 to cause growth defect. Interestingly Scd6 does not affect ability of Sbp1 to repress translation, which in turn does not affect Dhh1 and Pat1.
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Poornima, Gopalakrishna, Ravishankar Mythili, Priyabrata Nag, Sabnam Parbin, Praveen Kumar Verma, Tanweer Hussain i Purusharth I. Rajyaguru. "RGG-motif self-association regulates eIF4G-binding translation repressor protein Scd6". RNA Biology 16, nr 9 (12.06.2019): 1215–27. http://dx.doi.org/10.1080/15476286.2019.1621623.
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Roy, Debadrita, i Purusharth I. Rajyaguru. "Suppressor of clathrin deficiency (Scd6)-An emerging RGG-motif translation repressor". Wiley Interdisciplinary Reviews: RNA 9, nr 5 (22.05.2018): e1479. http://dx.doi.org/10.1002/wrna.1479.
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Lien, Pham Thi Kim, Keiichi Izumikawa, Kei Muroi, Kaoru Irie, Yasuyuki Suda i Kenji Irie. "Analysis of the Physiological Activities of Scd6 through Its Interaction with Hmt1". PLOS ONE 11, nr 10 (24.10.2016): e0164773. http://dx.doi.org/10.1371/journal.pone.0164773.
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Nocua, Paola A., José M. Requena i Concepción J. Puerta. "Identification of the interactomes associated with SCD6 and RBP42 proteins in Leishmania braziliensis". Journal of Proteomics 233 (luty 2021): 104066. http://dx.doi.org/10.1016/j.jprot.2020.104066.
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Bhatter, Nupur, Rajan Iyyappan, Gayatri Mohanan i Purusharth I. Rajyaguru. "Exploring the role of RRM domains and conserved aromatic residues in RGG motif of eIF4G-binding translation repressor protein Sbp1". Wellcome Open Research 3 (17.09.2021): 102. http://dx.doi.org/10.12688/wellcomeopenres.14709.3.
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Background: RNA binding proteins play crucial role in determining if a given mRNA will be translated, stored, or degraded. Sbp1 is an RGG-motif containing protein that is implicated in affecting mRNA decapping and translation. Sbp1 represses translation by binding eIF4G1 through its RGG-motif and activates decapping when overexpressed. In this report, we have assessed the genetic interaction of Sbp1 with decapping activators such as Dhh1, Pat1, and Scd6. We have further analyzed the importance of different domains and specific conserved residues of Sbp1 in its ability to cause over-expression mediated growth defect. Method: Sequence alignment was performed to identify conserved aromatic residues to be mutated. Using site-directed mutagenesis several point mutations and domain deletions were created in Sbp1 expressed under a galactose-inducible promoter. The mutants were tested for their ability to cause growth defect upon over-expression. The ability of Sbp1 to affect over-expression mediated growth defect of other decapping activators was tested using growth assay. Live cell imaging was done to study localization of Sbp1 and its RRM-deletion mutants to RNA granules upon glucose starvation. Results: Mutation of several aromatic residues in the RGG-motif and that of the phosphorylation sites in the RRM domain of Sbp1 did not affect the growth defect phenotype. Deletion of another eIF4G1-binding RGG-motif protein Scd6 does not affect the ability of Sbp1 to cause growth defect. Moreover, absence of Sbp1 did not affect the growth defect phenotypes observed upon overexpression of decapping activators Dhh1 and Pat1. Strikingly deletion of both the RRM domains (RRM1 and RRM2) and not the RNP motifs within them compromised the growth defect phenotype. Sbp1 mutant lacking both RRM1 and RRM2 was highly defective in localizing to RNA granules. Conclusion: This study identifies an important role of RRM domains independent of the RNP motif in Sbp1 function.
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Bhatter, Nupur, Rajan Iyyappan i Purusharth I. Rajyaguru. "Exploring the role of RRM domains and conserved aromatic residues in RGG motif of eIF4G-binding translation repressor protein Sbp1". Wellcome Open Research 3 (6.02.2020): 102. http://dx.doi.org/10.12688/wellcomeopenres.14709.2.
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Background: Mechanisms of mRNA fate decisions play an important role in determining if a given mRNA will be translated, stored or degraded upon arrival to cytoplasm. Sbp1 is an important RGG-motif containing protein that is implicated in affecting mRNA decapping and translation. Sbp1 represses translation by binding eIF4G1 through its RGG-motif and activates decapping when overexpressed. In this report we have assessed the genetic interaction of Sbp1 with decapping activators such as Dhh1, Pat1 and Scd6. We have further analyzed the importance of different domains and specific conserved residues of Sbp1 in translation repression activity. Method: Sequence alignment was performed to identify conserved aromatic residues to be mutated. Using site-directed mutagenesis several point mutations and domain deletions was created in Sbp1 expressed under a galactose-inducible promoter. The mutants were tested for their ability to cause growth defect upon over-expression. The ability of Sbp1 to affect over expression mediated growth defect of other decapping activators was tested using growth assay. Live cell imaging was done to study localization of Sbp1 and its RRM-deletion mutants to RNA granules upon glucose starvation. Results: Mutation of several aromatic residues in the RGG-motif and that of the phosphorylation sites in the RRM domain of Sbp1 did not affect the growth defect phenotype. Deletion of another eIF4G1-binding RGG-motif protein Scd6 does not affect the ability of Sbp1 to cause growth defect. Moreover, absence of Sbp1 did not affect the growth defect phenotypes observed upon overexpression of decapping activators Dhh1 and Pat1. Strikingly deletion of both the RRM domains (RRM1 and RRM2) and not the RNP motifs within them compromised the growth defect phenotype. Sbp1 mutant lacking both RRM1 and RRM2 was highly defective in localizing to RNA granules. Conclusion: This study identifies an important role of RRM domains independent of RNP motif in Sbp1 repression activity.
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Yadav, Sarita Ramsaran, Mangala Lakshmi Ragavan, Sanjeeb Kumar Mandal i Nilanjana Das. "DEGRADATION OF AZO DYE AND ELECTRICITY GENERATION USING YEAST MEDIATED MICROBIAL FUEL CELL". Fungal Territory 1, nr 1 (9.08.2018): 1–4. http://dx.doi.org/10.36547/ft.2018.1.1.1-4.
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In the present study, the efficiency of yeast mediated microbial fuel cell (MFC) was investigated towards degradation of Trypan blue (azo dye) and electricity generation. Five yeast strains viz. SC1, SC2, SCD1, SCD2, and SCD3 were isolated from different sources. The internal resistance of yeast isolates was tested using ferric oxide reduction method. To maximize the power density of MFC, NaCl was added to the medium and NaCl tolerance of yeast strains was tested. Among the five isolates, SC1 and SCD2 showed maximum ferric oxide reduction and NaCl tolerance. Initially, 5 % of SC1 and SCD2 yeast culture were inoculated in wastewater containing azo dye (100 µg/ml) in a H-type MFC chamber and 250 ml conical flask used as a control. Increased growth of yeast strain in MFC chamber was noted compared to conical flask culture. The data of electricity generation was taken for 15 days and electricity generation was measured using the multimeter. Maximum electricity generation was noted in SC1 (950mV) followed by SCD2 (750mV). In addition, SC1 could degrade azo dye more efficiently than SCD2. Therefore, it may be concluded thatSC1 yeast mediated MFC can be used as a potential technology for electricity generation and degradation of azo dye in wastewater.
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Olszewski, Jurek, Wiesław Chudzik, Kazimierz Wiśniewski, Jarosław Miłonski i Robert Matyja. "An Assessment of Concentrations of Soluble CD4 and CD8 Receptors in Serum before and after Surgical Treatment in Patients with Chronic Maxillary Sinusitis". American Journal of Rhinology 17, nr 3 (maj 2003): 123–26. http://dx.doi.org/10.1177/194589240301700301.
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Background The aim of this study was to assess the concentrations of soluble CD4 (sCD4) and sCD8 receptors in serum of patients before and after surgical treatment of chronic maxillary sinusitis. Methods We examined 57 patients, aged 20–63 years (mean age, 41 ± 0.5 years), and divided them into four groups: group I, 14 patients with chronic maxillary sinusitis without allergy; group II, 15 patients with chronic maxillary sinusitis with allergy; group III, 16 patients with cyst of maxillary sinuses without allergy (control); and group IV, 12 patients with cyst of maxillary sinuses with allergy (control). The assay of sCD4 and sCD8 receptor concentrations was performed by means of enzyme-linked immunosorbent assay method. The concentrations of sCD4 and sCD8 receptors before and after 30 days of surgical treatment of maxillary sinuses were examined. Results In our studies the increase of concentration of sCD4 in groups I and II in comparison with the concentration in control groups were statistically significant. The differences between mean concentrations of sCD8 in groups I and II and in the control groups were not statistically significant. After surgical treatment of chronic maxillary sinusitis, a significant decrease in values of sCD4 and sCD8 in comparison with the results before surgical treatment suggest that the measurement of cell suppression product concentration can be used to assess the extirpation of the inflammatory process and the effectiveness of the operation method. Conclusion Changes in concentration of sCD4 and sCD8 manifest activation or suppression of cells with particular receptor expression.
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Mnatsakanyan, Hayk, Caline Pechdimaljian, Roshani Jha, Alessandro Sammarco, Baolong Su, Kevin J. Williams, Steven J. Bensinger i Christian E. Badr. "CSIG-17. SCD5 PROTECTS GLIOBLASTOMA STEM CELLS FROM DEATH AND DIFFERENTIATION BY MODULATING INTRACELLULAR LIPID COMPOSITION". Neuro-Oncology 24, Supplement_7 (1.11.2022): vii42. http://dx.doi.org/10.1093/neuonc/noac209.166.
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Abstract Glioblastoma (GBM) is the most common malignant brain cancer in adults, enriched in a small subpopulation of glioma stem cells (GSC), which can drive tumor recurrence and therapeutic resistance. Considerable evidence suggests that the endogenous levels of unsaturated fatty acids (FA) are crucial regulators of GSCs survival and self-renewal. Stearoyl-CoA desaturase-1 (SCD-1) is the most abundant desaturase in humans. We have previously shown that SCD1 activity is required for GSCs self-renewal and brain tumor initiation. However, SCD1 orthologous isoform, SCD5, has been poorly characterized and its potential role in GBM has not been previously reported. We have observed that SCD5 is highly enriched in GSC both at the mRNA and protein levels. Genetic downregulation of SCD5 in GSCs led to a remarkable decrease in stem cell markers, impaired cell viability and the ability to form neurospheres. Further, the downregulation of SCD5 in GSCs orthotopically implanted in mice resulted in delayed tumor growth and extended overall survival. Shotgun lipidomics in GSCs after either SCD1 or SCD5 knock-down revealed a largely distinctive lipidome profile, highlighting the divergent role of these two isoforms in GBM lipid metabolism. Surprisingly, lipidomics analysis showed that both SCD1 and SCD5 are required to synthesize a variety of lipid species involved in receptor tyrosine kinase (RTKs) and GPCRs signal transduction, directly linking FA synthesis with the oncogenic signaling. We confirmed these results by immunoblot analysis. Using specific tagging and immunofluorescence analysis, we observed that, despite a spatial overlap in SCD1 and SCD5 expression, SCD5 is uniquely present in some subcellular locations. This suggests that different functions of these isoforms could be related to different subcellular localization. Altogether, our results underscore a novel function of SCD isoforms in GSCs metabolism and highlight SCD5 as a potential therapeutic target for GBM.
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Fromm, Simon A., Vincent Truffault, Julia Kamenz, Joerg E. Braun, Niklas A. Hoffmann, Elisa Izaurralde i Remco Sprangers. "The structural basis of Edc3- and Scd6-mediated activation of the Dcp1:Dcp2 mRNA decapping complex". EMBO Journal 31, nr 2 (15.11.2011): 279–90. http://dx.doi.org/10.1038/emboj.2011.408.
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Rajyaguru, Purusharth, Meipei She i Roy Parker. "Scd6 Targets eIF4G to Repress Translation: RGG Motif Proteins as a Class of eIF4G-Binding Proteins". Molecular Cell 45, nr 2 (styczeń 2012): 244–54. http://dx.doi.org/10.1016/j.molcel.2011.11.026.
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Chu, Dalena, Valeria Marrocco, Phoi Tiet, Jeanette Ampudia, Stephen Connelly i Cherie Ng. "Antigenic Modulation of CD6 By Itolizumab Is a New Mechanism for Effector T Cell Inhibition". Blood 138, Supplement 1 (5.11.2021): 995. http://dx.doi.org/10.1182/blood-2021-148805.
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Abstract Introduction Acute Graft-Versus-Host Disease (aGVHD) is a serious complication of hematopoietic stem cell transplantation (HSCT) that is primarily driven by alloreactive T cells. CD6 is a costimulatory receptor primarily expressed on T cells that promotes synapse formation, T cell activation, and migration into tissues by engaging with its ligand, activated leukocyte cell adhesion molecule (ALCAM). CD6 is expressed on reconstituting T cells soon after HSCT (Rambaldi et al., 2019) and early studies have shown that ex-vivo depletion of CD6 + donor cells prior to HSCT decreases the incidence of aGVHD (Soiffer et al., 1992; Soiffer et al., 1998). The reduced levels of aGVHD were attributed to an increased prevalence of CD6 - T cells that were less alloreactive (Rasmussen et al., 1994). Therefore, modulating activity of the CD6-ALCAM pathway may ameliorate aGVHD. Itolizumab is a humanized anti-CD6 monoclonal antibody that was previously described to block the engagement of ALCAM, thereby inhibiting T cell activity and trafficking. Here, we elucidate a second and highly novel mechanism in which antibody-mediated loss of CD6 from the surface of T cells results in CD6 low T cells that are hyporesponsive to T cell stimulation. Methods To assess the molecular mechanisms for itolizumab-induced loss of cell surface CD6, peripheral blood mononuclear cells (PBMCs) were exposed to different conditions including treatment with selected protease inhibitors, a membrane inhibitor, or Fc receptor blocking antibodies in the presence of itolizumab. Furthermore, B cells, NK cells, and monocytes were isolated from PBMCs and mixed with T cells in combination with itolizumab to evaluate cell contact requirements for loss. Following treatment, cell surface levels of CD6 protein were assessed by flow cytometry using a non-competing anti-CD6 detection antibody while analysis of full length CD6 was detected by western blot from total protein lysate. The soluble form of CD6 (sCD6) was analyzed by electrochemiluminescence and immunoprecipitation from supernatant of treated cells. Results Following treatment with itolizumab, the levels of cell surface CD6 was reduced as assessed by flow cytometry and western blot. Concurrent with a loss of cell surface CD6, levels of sCD6 in the cell supernatant increased, suggesting CD6 is primarily cleaved from the cell surface rather than internalized. Characterization of the cleaved sCD6 product by western blot revealed a 30KDa product. Itolizumab-induced changes in both levels of cell surface CD6 and sCD6 was abrogated by both 4-benzenesulfonyl fluoride hydrochloride (AEBSF), a serine protease inhibitor and cytochalasin D, an inhibitor of actin polymerization that prevents movement within the cell membrane and blocks endocytic trafficking. This suggests that a membrane-bound serine protease is responsible for cleavage. Itolizumab-induced CD6 cleavage did not occur with T cells alone and required the presence of other immune cells including monocytes and NK cells, but not B cells (Figure 1). In addition, when monocytes and T cells were separated by a membrane, cleavage of CD6 was not observed in the presence of itolizumab, indicating that cell-to-cell contact is required. Furthermore, blocking antibodies indicated that functional Fc receptors, especially FcγRIA, were required, suggesting that binding of itolizumab to both an Fc receptor on monocytes and CD6 on T cells predicated the cleavage event. The resulting CD6 low T cells were hyporesponsive to T cell stimulation, even in the absence of itolizumab, as indicated by reduced expression of PD-1, CD71 and CD25 as well as inflammatory cytokines (Figure 2). Inhibition was observed for both naïve and effector/memory T cell subsets. Conclusions Our results reveal a novel mechanism of antigenic modulation by itolizumab in which CD6 is cleaved from the T cell surface and released in a soluble form. Cleavage of CD6 occurs via a membrane-bound serine protease and appears dependent upon the engagement of itolizumab with Fc receptor(s) present on monocytes and NK cells. The loss of cell surface CD6 results in T cells with reduced responses to TCR-mediated stimulation. As such, treatment of aGVHD patients with itolizumab may reduce the alloreactivity of donor T cells, ameliorating disease symptoms and improving the clinical outcomes in these patients. Figure 1 Figure 1. Disclosures Chu: Equillium: Current Employment. Marrocco: Equillium, Inc.: Current Employment. Tiet: Equillium, Inc.: Current Employment. Ampudia: Equillium, Inc.: Current Employment. Connelly: Equillium: Current Employment, Divested equity in a private or publicly-traded company in the past 24 months, Membership on an entity's Board of Directors or advisory committees. Ng: Equillium: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months.
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Nascimento, Catarina, Andreia Gameiro, Jorge Correia, João Ferreira i Fernando Ferreira. "The Landscape of Tumor-Infiltrating Immune Cells in Feline Mammary Carcinoma: Pathological and Clinical Implications". Cells 11, nr 16 (18.08.2022): 2578. http://dx.doi.org/10.3390/cells11162578.
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Feline mammary carcinoma (FMC) shares key molecular and clinicopathological features with human breast cancer. We have herein studied the inflammatory infiltrate of FMC in order to uncover potential therapeutic targets and prognostic markers. To this end, the expression of different markers (CD3, CD4, CD8, CD20, CD56, FoxP3, CD68 and CD163) was analyzed in total, stromal (s) and intratumoral (i) tumor-infiltrating lymphocytes (TILs) and tumor-associated macrophages (TAMs), in 73 feline mammary carcinomas. The results revealed that higher percentages of sCD8+ TILs were associated with longer disease-free survival (p = 0.05) and overall survival (p = 0.021). Additionally, higher percentages of iCD4+ TILs correlated with positive lymph node status (p = 0.003), whereas CD163+ TAMs were associated with undifferentiated tumors (p = 0.013). In addition, sCD3+ (p = 0.033), sCD8+ (p = 0.044) and sCD68+ (p = 0.023) immune cells were enriched in triple negative normal-like carcinomas compared to other subtypes. Altogether, our results suggest that specific subsets of immune cells may play a major role in clinical outcome of cats with mammary carcinoma, resembling what has been reported in human breast cancer. These data further support the relevance of the feline model in breast cancer studies.
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Kogure, T., T. Itoh, Y. Shimada, T. Shintani, H. Ochiai i K. Terasawa. "Detection of serum soluble markers of immune activation in rheumatoid arthritis". Mediators of Inflammation 5, nr 4 (1996): 262–65. http://dx.doi.org/10.1155/s0962935196000373.
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The mutual correlation among soluble CD4 (sCD4), soluble CD8 (sCD8), and soluble CD23 (sCD23) has not yet been studied in patients with rheumatoid arthritis (RA), although previous studies have demonstrated that certain soluble markers of immune activation are elevated in RA. Thus, we examined this correlation based on the serum levels of sCD4, sCD8 and sCD23, and that of their levels with other serum markers such as immunoglobulin (Ig) subtypes (IgG, IgM and IgA), IgM-rheumatoid factor (IgM-RF) and C-reactive protein (CRP) in 25 RA patients, sCD4 was not elevated, whereas both sCD8 and sCD23 increased in RA patients compared with the healthy controls; a majority of RA patients, in particular, showed a high sCD23 level. The level of sCD23 showed a correlation with that of IgM-RF, but not with those of IgG, IgM, IgA and CRP. Importantly, a high level of sCD23 was not always accompartied with that of sCD8. The independent change between sCD23 and sCD8 levels was also observed in a one-year follow-up study of the two RA patients. These findings indicate that B cells might be generally activated in RA, whereas T-cell activation in variable in each patient with RA, suggesting that sCD23 is a more indicative marker for the immune status of RA patients than sCD8 from the clinical aspects.
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De Pablos, Luis Miguel, Steve Kelly, Janaina de Freitas Nascimento, Jack Sunter i Mark Carrington. "Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei". Open Biology 7, nr 4 (kwiecień 2017): 160159. http://dx.doi.org/10.1098/rsob.160159.
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The fate of an mRNA is determined by its interaction with proteins and small RNAs within dynamic complexes called ribonucleoprotein complexes (mRNPs). In Trypanosoma brucei and related kinetoplastids, responses to internal and external signals are mainly mediated by post-transcriptional processes. Here, we used proximity-dependent biotin identification (BioID) combined with RNA-seq to investigate the changes resulting from ectopic expression of RBP10 and RBP9, two developmentally regulated RNA-binding proteins (RBPs). Both RBPs have reduced expression in insect procyclic forms (PCFs) compared with bloodstream forms (BSFs). Upon overexpression in PCFs, both proteins were recruited to cytoplasmic foci, co-localizing with the processing body marker SCD6. Further, both RBPs altered the transcriptome from a PCF- to a BSF-like pattern. Notably, upon expression of BirA*-RBP9 and BirA*-RBP10, BioID yielded more than 200 high confidence protein interactors (more than 10-fold enriched); 45 (RBP9) and 31 (RBP10) were directly related to mRNA metabolism. This study validates the use of BioID for investigating mRNP components but also illustrates the complexity of mRNP function.
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Zeidan, Quira, Feng He, Fan Zhang, Hongen Zhang, Allan Jacobson i Alan G. Hinnebusch. "Conserved mRNA-granule component Scd6 targets Dhh1 to repress translation initiation and activates Dcp2-mediated mRNA decay in vivo". PLOS Genetics 14, nr 12 (7.12.2018): e1007806. http://dx.doi.org/10.1371/journal.pgen.1007806.
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Lamas, Iker, Nathalie Weber i Sophie G. Martin. "Activation of Cdc42 GTPase upon CRY2-Induced Cortical Recruitment Is Antagonized by GAPs in Fission Yeast". Cells 9, nr 9 (12.09.2020): 2089. http://dx.doi.org/10.3390/cells9092089.
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The small GTPase Cdc42 is critical for cell polarization in eukaryotic cells. In rod-shaped fission yeast Schizosaccharomyces pombe cells, active GTP-bound Cdc42 promotes polarized growth at cell poles, while inactive Cdc42-GDP localizes ubiquitously also along cell sides. Zones of Cdc42 activity are maintained by positive feedback amplification involving the formation of a complex between Cdc42-GTP, the scaffold Scd2, and the guanine nucleotide exchange factor (GEF) Scd1, which promotes the activation of more Cdc42. Here, we use the CRY2-CIB1 optogenetic system to recruit and cluster a cytosolic Cdc42 variant at the plasma membrane and show that this leads to its moderate activation also on cell sides. Surprisingly, Scd2, which binds Cdc42-GTP, is still recruited to CRY2-Cdc42 clusters at cell sides in individual deletion of the GEFs Scd1 or Gef1. We show that activated Cdc42 clusters at cell sides are able to recruit Scd1, dependent on the scaffold Scd2. However, Cdc42 activity is not amplified by positive feedback and does not lead to morphogenetic changes, due to antagonistic activity of the GTPase activating protein Rga4. Thus, the cell architecture is robust to moderate activation of Cdc42 at cell sides.
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Zeidan, Quira, Feng He, Fan Zhang, Hongen Zhang, Allan Jacobson i Alan G. Hinnebusch. "Correction: Conserved mRNA-granule component Scd6 targets Dhh1 to repress translation initiation and activates Dcp2-mediated mRNA decay in vivo". PLOS Genetics 15, nr 7 (23.07.2019): e1008299. http://dx.doi.org/10.1371/journal.pgen.1008299.
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Parbin, Sabnam, Gayatri Mohanan, Shirish Gole, Devika Joshi, Monmita Bhar i Purusharth I. Rajyaguru. "DEKTV and YVG motifs in the Lsm domain are important for the activity of Scd6, a conserved translation repressor protein". Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms 1863, nr 2 (luty 2020): 194474. http://dx.doi.org/10.1016/j.bbagrm.2019.194474.
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Levitsky, A., A. Gozhenko, V. Velichko i I. Selivanskaya. "The effect of dietary fat supplements on the activity of palmitic and stearic acid desaturases based on the results of a study of the fatty acid composition of neutral lipids in blood serum and liver of rats receiving a fat-free diet". Journal of Education, Health and Sport 12, nr 1 (18.01.2022): 197–206. http://dx.doi.org/10.12775/jehs.2022.12.01.016.
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Background. Desaturase enzymes are involved in the formation of monoenoic acids from saturated fatty acids. One such enzyme is stearyl-CoA-desaturase (SCD1), which converts stearic acid to oleic acid. The aim of this work was to determine the effect of edible fats with different fatty acid compositions on SCD1 activity.
Methods. High linoleic sunflower oil (HLSO), high oleic sunflower oil (HOSO) and palm oil (PO) were used. The rats were fed for 30 days with a semi-synthetic diet that did not contain any fats (FFD) and fat diets containing 5 % of each of the above oils. In animals, lipids were extracted from serum and liver and divided into 3 fractions: neutral lipids (NL), phospholipids (PL), and free fatty acids (FFA).
The fatty acid composition of each fraction was determined by gas chromatography. The SCD18 activity was determined by the C18:1 n-9/C18:0 ‒ ratio, and the SCD16 activity was determined by the C16:1 n-7/C16:0 ratio.
Results. A higher activity of SCD16 and SCD18 was found in the NL fraction, and the activity of SCD18 significantly exceeds that of SCD16. A decrease in the content of C16:0, C16:1 and C18:0 in the NL fraction of the liver and blood serum was shown. The activity of SCD16 in blood serum and liver decreases in rats fed fat diets, while the activity of SCD18 does not decrease, and even increases with the consumption of HOSO.
Conclusions. To determine the SCD1 activity, it is advisable to use the C18:1/C18:0 ratio in terms of the level of fatty acids in the NL fraction. Fatty diet inhibits SCD16 activity, and consumption of HOSO increases SCD18 activity.
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Kilchert, Cornelia, Julie Weidner, Cristina Prescianotto-Baschong i Anne Spang. "Defects in the Secretory Pathway and High Ca2+ Induce Multiple P-bodies". Molecular Biology of the Cell 21, nr 15 (sierpień 2010): 2624–38. http://dx.doi.org/10.1091/mbc.e10-02-0099.
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mRNA is sequestered and turned over in cytoplasmic processing bodies (PBs), which are induced by various cellular stresses. Unexpectedly, in Saccharomyces cerevisiae, mutants of the small GTPase Arf1 and various secretory pathway mutants induced a significant increase in PB number, compared with PB induction by starvation or oxidative stress. Exposure of wild-type cells to osmotic stress or high extracellular Ca2+ mimicked this increase in PB number. Conversely, intracellular Ca2+-depletion strongly reduced PB formation in the secretory mutants. In contrast to PB induction through starvation or osmotic stress, PB formation in secretory mutants and by Ca2+ required the PB components Pat1 and Scd6, and calmodulin, indicating that different stressors act through distinct pathways. Consistent with this hypothesis, when stresses were combined, PB number did not correlate with the strength of the translational block, but rather with the type of stress encountered. Interestingly, independent of the stressor, PBs appear as spheres of ∼40–100 nm connected to the endoplasmic reticulum (ER), consistent with the idea that translation and silencing/degradation occur in a spatially coordinated manner at the ER. We propose that PB assembly in response to stress occurs at the ER and depends on intracellular signals that regulate PB number.
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Iwaki, Aya, i Shingo Izawa. "Acidic stress induces the formation of P-bodies, but not stress granules, with mild attenuation of bulk translation in Saccharomyces cerevisiae". Biochemical Journal 446, nr 2 (14.08.2012): 225–33. http://dx.doi.org/10.1042/bj20120583.
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The stress response of eukaryotic cells often causes an attenuation of bulk translation activity and the accumulation of non-translating mRNAs into cytoplasmic mRNP (messenger ribonucleoprotein) granules termed cytoplasmic P-bodies (processing bodies) and SGs (stress granules). We examined effects of acidic stress on the formation of mRNP granules compared with other forms of stress such as glucose deprivation and a high Ca2+ level in Saccharomyces cerevisiae. Treatment with lactic acid clearly caused the formation of P-bodies, but not SGs, and also caused an attenuation of translation initiation, albeit to a lesser extent than glucose depletion. P-body formation was also induced by hydrochloric acid and sulfuric acid. However, lactic acid in SD (synthetic dextrose) medium with a pH greater than 3.0, propionic acid and acetic acid did not induce P-body formation. The results of the present study suggest that the assembly of yeast P-bodies can be induced by external conditions with a low pH and the threshold was around pH 2.5. The P-body formation upon acidic stress required Scd6 (suppressor of clathrin deficiency 6), a component of P-bodies, indicating that P-bodies induced by acidic stress have rules of assembly different from those induced by glucose deprivation or high Ca2+ levels.
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Moretti, Edgardo, Beatriz Basso, Liliana Cervetta, Ana Brigada i Gustavo Barbieri. "Patterns of Cytokines and Soluble Cellular Receptors in the Sera of Children with Acute Chagas' Disease". Clinical and Vaccine Immunology 9, nr 6 (listopad 2002): 1324–27. http://dx.doi.org/10.1128/cdli.9.6.1324-1327.2002.
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ABSTRACT Cytokines and soluble cellular receptors are involved in inflammatory processes and probably in the pathogenesis of parasite and bacterial diseases. In a previous study, we reported increased levels of soluble receptors of interleukin-2 (sIL2-R) in children with acute Chagas' disease, one of the main parasitic infections that is endemic in Latin America. We sought to analyze the pattern of different cytokines and soluble receptors in the sera of children with chagasic infection. Children with acute and indeterminate stages of Chagas' disease, as well as nonchagasic children, were studied. Sera were assayed by enzyme-linked immunosorbent assay to measure the levels of tumor necrosis factor alpha (TNF-α), IL-6, IL-2, IL-8, IL-12, sIL-2R, and the soluble receptors of CD8 and CD4 (sCD8 and sCD4). sIL-2R and sCD8 showed the highest levels in serum in acutely infected children, decreasing after specific antiparasite therapy. Chronic children showed a pattern similar to the one of nonchagasic children. Although they were not statistically significant, TNF-α, IL-6, and sCD4 showed a tendency to reach high levels in the acutely infected group, whereas IL-2, IL-8, and IL-12 did not reveal changes with respect to the noninfected children. In summary, we report here the patterns of cytokines and soluble receptors in in the sera of children infected with Trypanosoma cruzi; we found significantly increased levels of sIL-2R and sCD8 in acute infection that decreased after therapy, and high levels of TNF-α, IL-6, and sCD4 in some of the acute patients. The measurement of sIL-2R and sCD8 may provide a useful tool in the follow-up of children with Chagas' disease.
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Ladomery, Michael, i John Sommerville. "The Scd6/Lsm14 protein xRAPB has properties different from RAP55 in selecting mRNA for early translation or intracellular distribution in Xenopus oocytes". Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms 1849, nr 11 (listopad 2015): 1363–73. http://dx.doi.org/10.1016/j.bbagrm.2015.10.002.
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Ganner, Athina, Antonia Philipp, Simon Lagies, Laura Wingendorf, Lu Wang, Felicitas Pilz, Thomas Welte i in. "SCD5 Regulation by VHL Affects Cell Proliferation and Lipid Homeostasis in ccRCC". Cells 12, nr 6 (8.03.2023): 835. http://dx.doi.org/10.3390/cells12060835.
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Clear cell renal cell carcinoma (ccRCC) is the most common histological subtype of renal cancer, and inactivation of the VHL tumor suppressor gene is found in almost all cases of hereditary and sporadic ccRCCs. CcRCC is associated with the reprogramming of fatty acid metabolism, and stearoyl-CoA desaturases (SCDs) are the main enzymes controlling fatty acid composition in cells. In this study, we report that mRNA and protein expression of the stearoyl-CoA desaturase SCD5 is downregulated in VHL-deficient cell lines. Similarly, in C. elegans vhl-1 mutants, FAT-7/SCD5 activity is repressed, supporting an evolutionary conservation. SCD5 regulation by VHL depends on HIF, and loss of SCD5 promotes cell proliferation and a metabolic shift towards ceramide production. In summary, we identify a novel regulatory function of VHL in relation to SCD5 and fatty acid metabolism, and propose a new mechanism of how loss of VHL may contribute to ccRCC tumor formation and progression.
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Chu, Kiki, Makoto Miyazaki, Weng Chi Man i James M. Ntambi. "Stearoyl-Coenzyme A Desaturase 1 Deficiency Protects against Hypertriglyceridemia and Increases Plasma High-Density Lipoprotein Cholesterol Induced by Liver X Receptor Activation". Molecular and Cellular Biology 26, nr 18 (15.09.2006): 6786–98. http://dx.doi.org/10.1128/mcb.00077-06.
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ABSTRACT Stearoyl-coenzyme A desaturase (SCD) is the rate-limiting enzyme necessary for the biosynthesis of monounsaturated fatty acids. In this study, we investigated the regulation of mouse SCD1 by liver X receptor (LXR) and its role in plasma lipoprotein metabolism upon LXR activation. In vivo, the SCD1 gene remained induced upon LXR activation in the absence of sterol regulatory element-binding protein 1c (SREBP-1c), a known transcriptional regulator of SCD1. Serial deletion and point mutation analyses in reporter gene assays, as well as a gel mobility shift assay, identified an LXR response element in the mouse SCD1 promoter. In addition, SCD1 deficiency prevented the hypertriglyceridemic effect and reduced hepatic triglyceride accumulation associated with LXR activation despite induced hepatic expression of SREBP-1c protein and several SREBP1c and LXR target genes involved in lipoprotein metabolism. Unlike wild-type mice, SCD1-deficient mice failed to elevate the hepatic triglyceride monounsaturated acid (MUFA)/saturated fatty acid (SFA) ratio despite induction of the SCD2 gene. Together, these findings suggest that SCD1 plays a pivotal role in the regulation of hepatic and plasma triglyceride accumulation, possibly by modulating the MUFA-to-SFA ratio. In addition, SCD1 deficiency also increased plasma high-density lipoprotein cholesterol levels induced by LXR activation.
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Berthelot, V., L. Bernard, C. Richard, P. Chavatte-Palmer i Y. Heyman. "44 MUSCLE FATTY ACID COMPOSITION AND LIPOGENIC GENE EXPRESSION IN ADULT BOVINE CLONES AND CONTROL CATTLE". Reproduction, Fertility and Development 22, nr 1 (2010): 179. http://dx.doi.org/10.1071/rdv22n1ab44.
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Previous evaluation of milk and meat from clone cattle compared with AI control cows indicated that these products were in the normal range of data, but slight differences were observed in their fatty acid (FA) composition and muscle Δ9-desaturase indexes (Heyman et al. 2007 Animal 1, 936-972). It was therefore hypothesized that epigenic modifications induced by the nuclear transfer technology may affect the expression of the 2 genes [stearoyl-coenzyme A desaturase (SCD)-1, SCD5] responsible for Δ9-desaturation in bovines. The aim of the present experiment was to analyze the differences between clones and controls on FA composition and on SCD1 and SCD5 gene expression of the semitendinosus (ST) muscle. Biopsies of ST were taken from 5 clones from 2 different Holstein genotypes and 5 Holstein AI controls at 26 months of age. Each sample was immediately split into 3 aliquots, frozen in liquid nitrogen, and stored at -80°C until analysis. Fatty acid composition was analyzed by gas chromatography after lipid extraction and methylation according to Bas et al. (2005 Meat Sci. 71, 317-326). Total RNA was isolated from 300 mg of muscle tissue and abundance of SCD1 and SCD5 genes transcripts was determined by RT-PCRas described by Bernard et al. (2005 J. Dairy Sci. 88, 1478-1489) and Lengi and Corl (2007 Lipids 42, 499-508). Results are expressed as percentage of total FA for the FA composition and mRNA abundance of SCD1 and SCD5 determined as relative to the abundance of cyclophilin A mRNA. Statistical analyses were performed using the GLM procedure of SAS (SAS Institute, Cary, NC). Single degree of freedom orthogonal contrasts were used to compare effects of cloning (AI controls v. clones) as well as effects of clone genotypes (genotype 1 v. genotype 2). The C14:0, C14:1 cis-9, C16:0, C16:1 cis-9, and C18:0 proportions in ST were not different between clones and controls. However, clones tended to have a lower proportion of C18:1 n - 9 (-3.1% of total FA; P < 0.07) and higher proportions of C18:2 n - 6 (+1.2% of total FA; P < 0.01), C18:3 n - 3 (+0.7% of total FA; P < 0.05) and n - 3 polyunsaturated FA (+1.17% of total FA; P < 0.05) than controls. Ratios of C14 and C16 Δ9-desaturation in ST were not different between clones and controls but a lower C18 Δ9-desaturation ratio for the clones compared with controls was observed (0.76 v. 0.79; P < 0.05). The mRNA abundance of SCD1 was lower in clone compared with control cows (3.8 v. 8.5; P < 0.05), which could be explained by the higher proportion of n - 3 polyunsaturated FA observed in clones because of the negative effects of these polyunsaturated FA on SCD gene expression. The only difference observed between genotypes was for the C18:0 proportion in muscle (P < 0.05). In conclusion, in our set of animals, cloning decreased the ST muscle gene expression of SCD1 but not of SCD5, which is related to a slight decrease in C18 Δ9-desaturation ratio.
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Ho, AD, M. Maruyama, A. Maghazachi, JR Mason, S. Gluck i RE Corringham. "Soluble CD4, soluble CD8, soluble CD25, lymphopoieitic recovery, and endogenous cytokines after high-dose chemotherapy and blood stem cell transplantation". Blood 84, nr 10 (15.11.1994): 3550–57. http://dx.doi.org/10.1182/blood.v84.10.3550.3550.
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Abstract Mononuclear cell preparations from peripheral blood after mobilization with hematopoietic growth factors have been shown to induce accelerated neutrophil and platelet recovery as compared with that induced by autologous bone marrow transplantation after myeloablative chemotherapy. Because these mononuclear cell products contain many immunocompetent cells other than hematopoietic progenitors, these accessory cells might contribute to the rapid immunohematopoietic reconstitution. We have monitored the concentrations of soluble CD4 (sCD4), sCD8, and sCD25; the recovery of the lymphocyte subsets and of natural killer (NK) cells; and the endogenous levels of granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), IL-6, and granulocyte-macrophage-CSF (GM-CSF) in 12 patients who underwent high- dose chemotherapy supported by blood stem cells that were obtained by mobilization with chemotherapy and GM-CSF. The concentrations of both G- CSF and IL-6 peaked at 7 days after reinfusion of stem cells, and this transient elevation preceded the increase in the white blood cell count by approximately 5 to 7 days. The levels of sCD4 and sCD8 increased to a maximum on day 21, and the time to peak levels coincided with the maximum increase in white blood cell count, absolute neutrophil count, or lymphocytes. The levels of sCD25 were found to be elevated from day 7 to day 21. Statistically, the increases in sCD4, sCD8, sCD25, G-CSF, and IL-6 were highly significant, whereas there were no significant changes in IL-3 and GM-CSF. A rapid recovery of the NK activity was found in all 8 of the patients who could be monitored for this assay. Therefore, our study suggests that recovery of CD4+ cells, CD8+ cells, and NK activity coincided with that of neutrophils, which is preceded by a marked, but transient, elevation of IL-6 and G-CSF.
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Ho, AD, M. Maruyama, A. Maghazachi, JR Mason, S. Gluck i RE Corringham. "Soluble CD4, soluble CD8, soluble CD25, lymphopoieitic recovery, and endogenous cytokines after high-dose chemotherapy and blood stem cell transplantation". Blood 84, nr 10 (15.11.1994): 3550–57. http://dx.doi.org/10.1182/blood.v84.10.3550.bloodjournal84103550.
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Mononuclear cell preparations from peripheral blood after mobilization with hematopoietic growth factors have been shown to induce accelerated neutrophil and platelet recovery as compared with that induced by autologous bone marrow transplantation after myeloablative chemotherapy. Because these mononuclear cell products contain many immunocompetent cells other than hematopoietic progenitors, these accessory cells might contribute to the rapid immunohematopoietic reconstitution. We have monitored the concentrations of soluble CD4 (sCD4), sCD8, and sCD25; the recovery of the lymphocyte subsets and of natural killer (NK) cells; and the endogenous levels of granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), IL-6, and granulocyte-macrophage-CSF (GM-CSF) in 12 patients who underwent high- dose chemotherapy supported by blood stem cells that were obtained by mobilization with chemotherapy and GM-CSF. The concentrations of both G- CSF and IL-6 peaked at 7 days after reinfusion of stem cells, and this transient elevation preceded the increase in the white blood cell count by approximately 5 to 7 days. The levels of sCD4 and sCD8 increased to a maximum on day 21, and the time to peak levels coincided with the maximum increase in white blood cell count, absolute neutrophil count, or lymphocytes. The levels of sCD25 were found to be elevated from day 7 to day 21. Statistically, the increases in sCD4, sCD8, sCD25, G-CSF, and IL-6 were highly significant, whereas there were no significant changes in IL-3 and GM-CSF. A rapid recovery of the NK activity was found in all 8 of the patients who could be monitored for this assay. Therefore, our study suggests that recovery of CD4+ cells, CD8+ cells, and NK activity coincided with that of neutrophils, which is preceded by a marked, but transient, elevation of IL-6 and G-CSF.
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Putz, D., U. Barnas, A. Luger, G. Mayer, W. Woloszczuk i H. Graf. "Biocompatibility of High-Flux Membranes". International Journal of Artificial Organs 15, nr 8 (sierpień 1992): 456–60. http://dx.doi.org/10.1177/039139889201500802.
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Standard dialysis with cuprophane membranes is known to stimulate the immune system. As a result of activation of macrophages various interleukins and tumor necrosis factor (TNF) are secreted, presenting further evidence of the poor biocompatibility of cuprophane. We investigated the immunogenic properties of three modern high-flux membranes. Seven patients were studied during hemodiafiltration sessions using either a polysulfone (F60, Fresenius), a polymethylmetacrylate (BK 2.1, Toray) or a cellulose triacetate (FB-210 U, Nipro) dialyzer in a hemodiafiltration procedure. Serial measurements were made during each treatment of interleukin-1β (II-1β), TNF, soluble IL-2 receptor (sII-2r), soluble CD4 (sCD4), soluble CD8 (sCD8), interferon gamma (IFNg) and neopterin. In contrast to the known increase of IL-1β, IL-2r and TNF with cuprophane membranes, none of the modern high-flux dialyzers stimulated the production of these factors. Significant decreases of neopterin and sCD4 were observed. IFNg and sCD8 did not change significantly. Our results suggest that the modern high-flux dialyzers are non-immunogenic, and thus provide further evidence of the superior biocompatibility of synthetic or semisynthetic membranes over the conventional cuprophane.
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Boguszewska, Karolina, Michał Szewczuk, Julia Kaźmierczak-Barańska i Bolesław T. Karwowski. "How (5′S) and (5′R) 5′,8-Cyclo-2′-Deoxypurines Affect Base Excision Repair of Clustered DNA Damage in Nuclear Extracts of xrs5 Cells? A Biochemical Study". Cells 10, nr 4 (24.03.2021): 725. http://dx.doi.org/10.3390/cells10040725.
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The clustered DNA lesions (CDLs) are a characteristic feature of ionizing radiation’s impact on the human genetic material. CDLs impair the efficiency of cellular repair machinery, especially base excision repair (BER). When CDLs contain a lesion repaired by BER (e.g., apurinic/apyrimidinic (AP) sites) and a bulkier 5′,8-cyclo-2′-deoxypurine (cdPu), which is not a substrate for BER, the repair efficiency of the first one may be affected. The cdPus’ influence on the efficiency of nuclear BER in xrs5 cells have been investigated using synthetic oligonucleotides with bi-stranded CDL (containing (5′S) 5′,8-cyclo-2′-deoxyadenosine (ScdA), (5′R) 5′,8-cyclo-2′-deoxyadenosine (RcdA), (5′S) 5′,8-cyclo-2′-deoxyguanosine (ScdG) or (5′R) 5′,8-cyclo-2′-deoxyguanosine (RcdG) in one strand and an AP site in the other strand at different interlesion distances). Here, for the first time, the impact of ScdG and RcdG was experimentally tested in the context of nuclear BER. This study shows that the presence of RcdA inhibits BER more than ScdA; however, ScdG decreases repair level more than RcdG. Moreover, AP sites located ≤10 base pairs to the cdPu on its 5′-end side were repaired less efficiently than AP sites located ≤10 base pairs on the 3′-end side of cdPu. The strand with an AP site placed opposite cdPu or one base in the 5′-end direction was not reconstituted for cdA nor cdG. CdPus affect the repair of the other lesion within the CDL. It may translate to a prolonged lifetime of unrepaired lesions leading to mutations and impaired cellular processes. Therefore, future research should focus on exploring this subject in more detail.
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Rincon, Gonzalo, Alma Islas-Trejo, Alejandro R. Castillo, Dale E. Bauman, Bruce J. German i Juan F. Medrano. "Polymorphisms in genes in the SREBP1 signalling pathway and SCD are associated with milk fatty acid composition in Holstein cattle". Journal of Dairy Research 79, nr 1 (25.11.2011): 66–75. http://dx.doi.org/10.1017/s002202991100080x.
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Genes in the sterol regulatory element-binding protein-1 (SREBP1) pathway play a central role in regulation of milk fat synthesis, especially the de-novo synthesis of saturated fatty acids. SCD, a SREBP-responsive gene, is the key enzyme in the synthesis of monounsaturated fatty acids in the mammary gland. In the present study, we discovered SNP in candidate genes associated with this signalling pathway and SCD to identify genetic markers that can be used for genetic and metabolically directed selection in cattle. We resequenced six candidate genes in the SREBP1 pathway (SREBP1, SCAP, INSIG1, INSIG2, MBTPS1, MBTPS2) and two genes for SCD (SCD1 and SCD5) and discovered 47 Tag SNP that were used in a marker-trait association study. Milk and blood samples were collected from Holstein cows in their 1st or 2nd parity at 100–150 days of lactation. Individual fatty acids from C4 to C20, saturated fatty acid (SFA) content, monounsaturated fatty acid content, polyunsaturated fatty acid content and desaturase indexes were measured and used to perform the asociation analysis. Polymorphisms in the SCD5 and INSIG2 genes were the most representative markers associated with SFA/unsaturated fatty acid (UFA) ratio in milk. The analysis of desaturation activity determined that markers in the SCD1 and SCD5 genes showed the most significant effects. DGAT1 K232A marker was included in the study to examine the effect of this marker on the variation of milk fatty acids in our Holstein population. The percentage of variance explained by DGAT1 in the analysis was only 6% of SFA/UFA ratio. Milk fat depression was observed in one of the dairy herds and in this particular dairy one SNP in the SREBP1 gene (rs41912290) accounted for 40% of the phenotypic variance. Our results provide detailed SNP information for key genes in the SREBP1 signalling pathway and SCD that can be used to change milk fat composition by marker-assisted breeding to meet consumer demands regarding human health, as well as furthering understanding of technological aspects of cows' milk.
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Kuryliszyn-Moskal, A., K. Bemacka i O. Bielecka. "Soluble CD4 (sCD4), CD8 (sCD8) and cytokine levels in rheumatoid arthritis complicated by vasculitis". Immunology Letters 56 (maj 1997): 321. http://dx.doi.org/10.1016/s0165-2478(97)86296-7.
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Kuryliszyn-Moskal, A. "Soluble CD4 (sCD4), CD8 (sCD8) and cytokine levels in rheumatoid arthritis complicated by vasculitis". Immunology Letters 56, nr 1-3 (maj 1997): 321. http://dx.doi.org/10.1016/s0165-2478(97)88134-5.
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Tabor, D. E., Y. R. Xia, M. Mehrabian, P. A. Edwards i A. J. Lusis. "A cluster of stearoyl CoA desaturase genes, Scd1 and Scd2, on mouse Chromosome 19". Mammalian Genome 9, nr 4 (kwiecień 1998): 341–42. http://dx.doi.org/10.1007/s003359900765.
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ZHANG, Shaobo, Yanzhu YANG i Yuguang SHI. "Characterization of human SCD2, an oligomeric desaturase with improved stability and enzyme activity by cross-linking in intact cells". Biochemical Journal 388, nr 1 (10.05.2005): 135–42. http://dx.doi.org/10.1042/bj20041554.
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SCD (stearoyl-CoA desaturase) catalyses the conversion of saturated fatty acids into mono-unsaturated fatty acids, a critical step involved in lipid metabolism and various other biological functions. In the present study, we report the identification and characterization of a human gene that encodes a novel SCD enzyme (hSCD2). The hSCD2 gene codes for a 37.5-kDa protein that shares 61% and 57% sequence identity with the human SCD1 and mouse SCD2 enzymes respectively. The recombinant hSCD2 enzyme expressed in mammalian and Sf9 insect cells efficiently catalysed desaturation of both stearoyl- and palmitoyl-CoAs to the corresponding mono-unsaturated fatty acids. In comparison with the hSCD1 gene that is predominantly expressed in liver, hSCD2 is most abundantly expressed in pancreas and brain. Additionally, hSCD2 transcripts from adult and foetal tissues exhibit different sizes because of alternative splicing in the non-coding region, suggesting that hSCD2 expression is developmentally regulated. The recombinant human SCD2 and SCD1 transiently expressed in COS-7 cells exhibited as oligomeric proteins that consist of homodimers and oligomers when resolved by SDS/PAGE. The complex formation was independent of SCD protein expression levels, as supported by a relatively constant ratio of the level of dimers and oligomers to that of the monomers from COS-7 cells transiently transfected with different amounts of SCD expression vectors. Furthermore, treatment of intact COS-7 cells with a cross-linking reagent resulted in dose-dependent increases in the levels of SCD protein and activity, suggesting that oligomerization may play an important role in regulating the stability of SCD enzymes.
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Green, Christopher D., i L. Karl Olson. "Modulation of palmitate-induced endoplasmic reticulum stress and apoptosis in pancreatic β-cells by stearoyl-CoA desaturase and Elovl6". American Journal of Physiology-Endocrinology and Metabolism 300, nr 4 (kwiecień 2011): E640—E649. http://dx.doi.org/10.1152/ajpendo.00544.2010.
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Induction of endoplasmic reticulum (ER) stress and apoptosis by elevated exogenous saturated fatty acids (FAs) plays a role in the pathogenesis of β-cell dysfunction and loss of islet mass in type 2 diabetes. Regulation of monounsaturated FA (MUFA) synthesis through FA desaturases and elongases may alter the susceptibility of β-cells to saturated FA-induced ER stress and apoptosis. Herein, stearoyl-CoA desaturase (SCD)1 and SCD2 mRNA expression were shown to be induced in islets from prediabetic hyperinsulinemic Zucker diabetic fatty (ZDF) rats, whereas SCD1, SCD2, and fatty acid elongase 6 (Elovl6) mRNA levels were markedly reduced in diabetic ZDF rat islets. Knockdown of SCD in INS-1 β-cells decreased desaturation of palmitate to MUFA, lowered FA partitioning into complex neutral lipids, and increased palmitate-induced ER stress and apoptosis. Overexpression of SCD2 increased desaturation of palmitate to MUFA and attenuated palmitate-induced ER stress and apoptosis. Knockdown of Elovl6 limited palmitate elongation to stearate, increasing palmitoleate production and attenuating palmitate-induced ER stress and apoptosis, whereas overexpression of Elovl6 increased palmitate elongation to stearate and palmitate-induced ER stress and apoptosis. Overall, these data support the hypothesis that enhanced MUFA synthesis via upregulation of SCD2 activity can protect β-cells from elevated saturated FAs, as occurs in prediabetic states. Overt type 2 diabetes is associated with diminished islet expression of SCD and Elovl6, and this can disrupt desaturation of saturated FAs to MUFAs, rendering β-cells more susceptible to saturated FA-induced ER stress and apoptosis.
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ZHANG, Lin, Lan GE, Tai TRAN, Kurt STENN i Stephen M. PROUTY. "Isolation and characterization of the human stearoyl-CoA desaturase gene promoter: requirement of a conserved CCAAT cis-element". Biochemical Journal 357, nr 1 (25.06.2001): 183–93. http://dx.doi.org/10.1042/bj3570183.
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Stearoyl-CoA desaturase is the rate-limiting enzyme in the production of mono-unsaturated fatty acids. We have recently cloned and characterized the human Scd cDNA and SCD (the stearoyl-CoA desaturase structural gene) on chromosome 10, as well as the non-transcribed pseudogene on chromosome 17. In order to further define SCD regulation and function, we have isolated and characterized the promoter of the structural gene. Screening of chromosome-10-specific libraries resulted in the isolation of 4.1kb of SCD sequence upstream of the translation start site. Binding sites for transcription factors critical for mouse Scd1 and Scd2 promoter activity, such as sterol-regulated-element-binding protein and nuclear factor Y, were present in the human SCD promoter (Scd is the mouse stearoyl-CoA desaturase gene). Deletion analysis in HaCaT keratinocytes identified a critical region for promoter activity between nts 496–609 upstream of the translation start site. Site-directed mutagenesis of binding sites in this region identified the CCAAT box as the critical cis-element for SCD promoter activity. An electrophoretic mobility-shift assay confirmed that this element binds nuclear proteins from HaCaT keratinocytes. The polyunsaturated-fatty-acid (PUFA) response element, previously identified in the promoters of mouse Scd1 and Scd2, was found to be conserved in the human SCD promoter, and contained the critical CCAAT cis-element. A minimal promoter construct including this region was responsive to fatty acids, with oleate and linoleate decreasing transcription and stearate increasing it. These studies indicate that CCAAT-box-binding proteins activate SCD transcription in cultured keratinocytes and that fatty acids modulate transcription, most likely through the conserved PUFA response element.
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WHEATLEY, Edward, i Katrin RITTINGER. "Interactions between Cdc42 and the scaffold protein Scd2: requirement of SH3 domains for GTPase binding". Biochemical Journal 388, nr 1 (10.05.2005): 177–84. http://dx.doi.org/10.1042/bj20041838.
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The multi-domain protein Scd2 acts as a scaffold upon which the small GTPase Cdc42 (cell division cycle 42), its nucleotide-exchange factor Scd1 and the p21-activated kinase Shk1 assemble to regulate cell polarity and the mating response in fission yeast. In the present study, we show using isothermal titration calorimetry that Scd2 binds two molecules of active GTP-bound Cdc42 simultaneously, but independently of one another. The two binding sites have significantly different affinities, 21 nM and 3 μM, suggesting that they play distinct roles in the Shk1 signalling network. Each of the Cdc42-binding sites includes one of the SH3 (Src homology 3) domains of Scd2. Our data indicate that complex formation does not occur in a conventional manner via the conserved SH3 domain ligand-binding surface. Neither of the isolated SH3 domains is sufficient to interact with the GTPase, and they both require adjacent regions to either stabilize their conformations or contribute to the formation of the Cdc42-binding surface. Furthermore, we show that there is no evidence for an intramolecular PX–SH3 domain interaction, which could interfere with SH3 domain function. This work suggests that SH3 domains might contribute directly to signalling through small GTPases and thereby adds another aspect to the diverse nature of SH3 domains as protein–protein-interaction modules.
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De Rie, MA, IM Zonneveld, L. Witkamp, RA Van Lier, TA Out i JD Bos. "Soluble interleukin-2 receptor (sIL-2R) is a marker of disease activity in psoriasis: a comparison of sIL-2R, sCD27, sCD4, sCD8 and sICAM-1." Acta Dermato-Venereologica 76, nr 5 (1.09.1996): 357–60. http://dx.doi.org/10.2340/0001555576357360.
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Psoriasis is a T-cell-mediated inflammatory skin disease which can be treated successfully with immunosuppressive drugs. Our purpose was to evaluate disease activity of psoriasis and the effect of immunosuppressive treatment by monitoring the soluble T-cell products sIL-2R, sCD27, sCD4, sCD8 and sICAM-1. Twenty-two patients were treated orally with escalating dosages of cyclosporin A (n = 17)(3-5 mg/kg/day) or FK506 (n = 5)(0.05-0.15 mg/kg/day). The Psoriasis Area and Severity Index (PASI) was used to monitor clinical activity of psoriasis. Serum samples were analyzed by ELISA. sIL-2R levels showed the highest correlation with psoriasis disease activity (rs = 0.89
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Scott, Julia S., Reuben Young, Swati Irani, Jonas Dehairs, Stephen Blanksby, Johannes V. Swinnen, Zeyad D. Nassar i Lisa M. Butler. "Abstract A031: A fat lot of good: A novel monounsaturated fatty acid promotes prostate cancer growth and survival". Cancer Research 83, nr 11_Supplement (2.06.2023): A031. http://dx.doi.org/10.1158/1538-7445.prca2023-a031.
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Abstract Advanced prostate cancer (PCa) is currently incurable, and development of novel treatments requires a greater understanding of key pathways that are altered during PCa progression. We have focussed on fatty acid metabolism, as the major source of energy for PCa cells, and have implicated a monounsaturated fatty acid (MUFA), cis-vaccenic acid (cVA), as a critical regulator of PCa cell homeostasis. Production of cVA requires stearoyl-CoA desaturase 1 (SCD1), the enzyme attributed to general MUFA production, and whose activity is important in a range of cancer types. Nonetheless, the specific role of cVA has not previously been studied, in PCa nor cancer more generally. To elucidate the role of cVA in PCa more precisely, we used SCD1 inhibition to study PCa responses to broad MUFA depletion and specific MUFA supplementation, comparing the responses of cVA with its more common structural isomer oleate. To accomplish these aims, we utilised cell line models of PCa (LNCaP, MR49F and V16D) and a benign prostate cell line (PNT1A) to perform cell viability, lipidomic and flow cytometric assays. In addition, responses in clinically-relevant patient-derived tumor explants were measured by proliferative (Ki67) and cell death (CC3) marker staining. Pharmacological SCD1 inhibition (SCDi; A939572) decreased cell viability and increased cell death in PCa cell lines and patient-derived tumor explants, while only minimally affecting the benign prostate cell line PNT1A. Phenotypically, analysis of mitochondrial function and lipidomic changes under SCDi suggested a role for MUFAs in regulating mitochondrial membrane composition via cardiolipins, and oxidative stress levels. This in turn has the potential to regulate cell death, control tumor burden, and influence overall PCa patient survival. Supplementation of cVA or oleate under SCDi demonstrated that only cVA was successful at rescuing cell viability, and cVA, but not oleate, could rescue changes observed in mitochondrial-related lipids, suggesting that cVA regulation of mitochondrial function was a major determinant of its effect on cell viability. Additionally, cVA alone dose-dependently increased PCa cell viability, implicating cVA as an important oncogenic factor. Together, these data identify cVA as a novel substrate required for PCa cell viability, likely via regulation of mitochondrial homeostasis and associated cell death pathways. In summary, this research has revealed more precise insights into the oncogenic effects of an overlooked MUFA, cVA, in PCa, and may uncover new druggable targets that are more selective, to improve treatment options for advanced PCa. Citation Format: Julia S. Scott, Reuben Young, Swati Irani, Jonas Dehairs, Stephen Blanksby, Johannes V. Swinnen, Zeyad D. Nassar, Lisa M. Butler. A fat lot of good: A novel monounsaturated fatty acid promotes prostate cancer growth and survival [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A031.
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Yin, Xin-Ke, Chao Wang, Li-Li Feng, Shao-Mei Bai, Wei-Xing Feng, Neng-Tai Ouyang, Zhong-Hua Chu, Xin-Juan Fan i Qi-Yuan Qin. "Expression Pattern and Prognostic Value of CTLA-4, CD86, and Tumor-Infiltrating Lymphocytes in Rectal Cancer after Neoadjuvant Chemo(radio)therapy". Cancers 14, nr 22 (14.11.2022): 5573. http://dx.doi.org/10.3390/cancers14225573.
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The synergistic effect of combining immune checkpoint inhibitors (ICIs) with neoadjuvant chemo(radio)therapy (nCRT) in colorectal cancer is still limited. We aimed to understand the impact of nCRT on the tumor microenvironment and to explore favorable immune markers of this combination. Herein, we investigated the expression of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), CD86, CD4, and CD8 after nCRT and its association with clinicopathological characteristics. Immunostaining of immune-related molecules was performed in 255 surgically resected specimens from rectal cancer patients treated with nCRT. CD4 and CD8 expression on the tumor (tCD4/CD8), stroma (sCD4/CD8), and invasive front (iCD4/CD8) was evaluated. The expression levels of immune-related molecules were significantly lower in the nCRT-treated group, except for CTLA-4 and sCD8. However, patients with higher sCD8+ cell density and CTLA-4 expression had better progression-free survival (PFS) and distant metastasis-free survival (DMFS). In addition, higher CD86 expression was associated with poorer overall survival (OS). Higher CTLA-4 expression was associated with higher tCD8+ cell density, whereas CD86 expression was correlated with the cell density of t/sCD8. Prognostic analysis confirmed that the relationships between CTLA-4 and DMFS as well as CD86 and OS were significantly correlated in low rather than high CD8+ cell density. Further the combination of CD8+ cell density and CD86 expression was shown to be an independent prognostic factor of OS, whereas the combination of CTLA-4 was not for DMFS. Together, these results demonstrate significant correlations between CD86 expression and t/sCD8+ cell density in rectal cancer after nCRT and could potentially have clinical implications for combining ICIs and nCRT.
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Huang, K. M., L. Gullberg, K. K. Nelson, C. J. Stefan, K. Blumer i S. K. Lemmon. "Novel functions of clathrin light chains: clathrin heavy chain trimerization is defective in light chain-deficient yeast". Journal of Cell Science 110, nr 7 (1.04.1997): 899–910. http://dx.doi.org/10.1242/jcs.110.7.899.
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Clathrin is a major coat protein involved in sorting and retention of proteins at the late Golgi and in endocytosis from the cell surface. The clathrin triskelion contains three heavy chains, which provide the structural backbone of the clathrin lattice and three light chains, which are thought to regulate the formation or disassembly of clathrin coats. To better understand the function of the clathrin light chain, we characterized yeast strains carrying a disruption of the clathrin light chain gene (CLC1). Light chain-deficient cells showed phenotypes similar to those displayed by yeast that have a disruption in the clathrin heavy chain gene (CHC1). In clc1-delta cells, the steady state level of the clathrin heavy chain was reduced to 20%-25% of wild-type levels and most of the heavy chain was not trimerized. If CHC1 was overexpressed in clc1-delta cells, heavy chain trimers were detected and several clc1-delta phenotypes were partially rescued. These results indicate that the light chain is important for heavy chain trimerization and the heavy chain still has some function in the absence of the light chain. In yeast, deletion of CHC1 is lethal in strains carrying the scd1-i allele, while strains carrying the scd1-v allele can survive without the heavy chain. In previous studies we isolated several multicopy suppressors of inviability of chc1-delta scd1-i cells. Surprisingly, one of these suppressors, SCD4, is identical to CLC1. Overexpression of CLC1 in viable chc1-delta scd1-v strains rescued some but not all of the phenotypes displayed by these cells. In the absence of the heavy chain, the light chain was not found in a high molecular mass complex, but still associated with membranes. These results suggest that the light chain can function independently of the clathrin heavy chain in yeast.
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Mayers, Jonathan Russell, Tianwei Hu, Chao Wang, Jessica J. Cárdenas, Yuqi Tan, Jianwei Pan i Sebastian Y. Bednarek. "SCD1 and SCD2 Form a Complex That Functions with the Exocyst and RabE1 in Exocytosis and Cytokinesis". Plant Cell 29, nr 10 (29.09.2017): 2610–25. http://dx.doi.org/10.1105/tpc.17.00409.
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