Relevant bibliographies by topics / In-Cell EPR

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Academic literature on the topic 'In-Cell EPR'

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Published: 25 May 2024

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Journal articles on the topic "In-Cell EPR"

Adida, C., G. Ambrosini, J. Plescia, PL Crotty, J. Costa, and DC Altieri. "Protease receptors in Hodgkin's disease: expression of the factor Xa receptor, effector cell protease receptor-1, in Reed-Sternberg cells." Blood 88, no. 4 (August 15, 1996): 1457–64. http://dx.doi.org/10.1182/blood.v88.4.1457.bloodjournal8841457.

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The expression of a cellular receptor for the blood-clotting protease factor Xa, designated effector cell protease receptor-1 (EPR-1), was investigated in lymphoma. Immunohistochemical analysis demonstrated prominent reactivity of monoclonal antibodies to EPR-1 with Reed- Sternberg cells in 30 of 35 cases of nodular-sclerosis, lymphocyte- depletion, and mixed-cellularity Hodgkin's disease (HD). In contrast, several non-Hodgkin's lymphomas, or the nonneoplastic cellular components of HD, did not react with anti-EPR-1 monoclonal antibodies. A single molecular species of approximately 62 kD, consistent with the size and structural organization of EPR-1, was immunoblotted by an anti- EPR-1 monoclonal antibody from tissue samples of HD, but not from normal lymph nodes. Expression of EPR-1 transcripts in Reed-Sternberg cells was demonstrated by in situ hybridization with an antisense EPR-1 riboprobe, and by amplification of reverse-transcribed HD RNA with EPR- 1-specific primers. These findings identify the factor Xa receptor, EPR- 1, as a novel marker of Reed-Sternberg cells, and suggest its potential role in the histopathogenesis of HD.

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Cattani, Julia, Vinod Subramaniam, and Malte Drescher. "Room-temperature in-cell EPR spectroscopy: alpha-Synuclein disease variants remain intrinsically disordered in the cell." Physical Chemistry Chemical Physics 19, no. 28 (2017): 18147–51. http://dx.doi.org/10.1039/c7cp03432f.

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Hänsel, Robert, Laura M. Luh, Ivan Corbeski, Lukáš Trantirek, and Volker Dötsch. "In-Cell NMR and EPR Spectroscopy of Biomacromolecules." Angewandte Chemie International Edition 53, no. 39 (July 28, 2014): 10300–10314. http://dx.doi.org/10.1002/anie.201311320.

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Piknova, Barbora, Mark T. Gladwin, Cheryl A. Hillery, and Neil Hogg. "Electron Paramagnetic Resonance Study of Cell-Free Hemoglobin in Sickle Cell Disease: Potential Antioxidant Role of Haptoglobin." Blood 106, no. 11 (November 16, 2005): 2345. http://dx.doi.org/10.1182/blood.v106.11.2345.2345.

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Abstract It has previously been shown that red blood cell-free oxyhemoglobin (oxyHb) can affect endothelial function in Sickle Cell Disease (SCD) by virtue of its NO scavenging properties (Reiter et al, Nat Med, 2002). This reaction may have down-stream consequence via formation of metHb which has been demonstrated to have pro-oxidant properties and can oxidize low-density lipoprotein in vitro. In this study we have examined the fate of metHb in SCD vs normal plasma by examining the characteristic single line EPR signature of metHb in the g = 6 region. We show that the EPR signature of metHb differs in SCD compared with normals and exhibits a split EPR line indicative of a more rhombic heme geometry (See Figure). Figure Figure Addition of purified metHb to SCD plasma results in a conversion of the single line EPR spectrum to the rhombic form. By addition of hemin and hemoglobin to plasma constituents we have determined that the rhombic EPR spectrum is consistent with heme bound to serum albumin. In addition, immunoprecipitation of serum albumin from SCD plasma with EPR analysis of the IP fraction revealed the presence of albumin associated metHb. Interestingly, the transfer of heme from metHb to albumin is accelereated by the presence of low-density lipoprotein and inhibited by the addition of haptoglobin suggesting that plasma components can modulate heme release from metHb and that the lack of haptoglobin in SCD plasma allowed the transfer of heme from metHb to albumin. To confirm this we added metHb to SCD plasma in the presence and absence of haptoglobin and observed that the addition of haptoglobin prevented conversion of the metHb spectrum to the albumin associated split (rhombic) species. From these studies we conclude that plasma haptoglobin not only binds hemoglobin but also limits heme release from metHb. To examine potential oxidative consequences of this effect, we incubated LDL with metHb in the presence and absence of haptoglobin. Haptoglobin inhibited LDL oxidation in a dose-dependent manner. Interestingly, the 1-1 isoform of haptoglobin was much more effective than the 2-2 isoform, in agreement with published studies on the efficacy of hemoglobin binding to the two haptoglobin varieties. Additionally, we have measured levels of 8-isoprostanes in the plasma of SCD patients as an index of lipid oxidation and show that 8-isoprostane levels are significantly elevated in SCD patients (295 209 pg/ml, mean S.D., n= 13, vs. 69 37 pg/ml, mean S.D. n= 5 for normal controls). In conclusion, these studies provide evidence that the saturation of the haptoglobin clearance mechanisms in hemolytic disorders such as SCD can not only affect the NO-axis of endothelial function due to increases in cell-free hemoglobin, but can also lead to altered heme metabolism leading to lipid oxidation and formation of pro-inflammatory lipids which have the potential to alter both endothelial and red cell function.

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Zaman, Guido J. R., and Edward M. Conway. "The elusive factor Xa receptor: failure to detect transcripts that correspond to the published sequence of EPR-1." Blood 96, no. 1 (July 1, 2000): 145–48. http://dx.doi.org/10.1182/blood.v96.1.145.

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Abstract The coagulation protease factor Xa induces cellular responses implicated in cardiovascular and inflammatory disease. Effector-cell protease receptor 1 (EPR-1) is a functionally characterized receptor of factor Xa, and the EPR-1complementary DNA (cDNA) was published. Remarkably, the cDNA encoding an inhibitor of apoptosis, survivin, is reportedly identical to that ofEPR-1 except for a few nucleotide differences and its orientation opposite to EPR-1. To isolate the EPR-1cDNA and gene, we surveyed gene databases for expressed sequence tags (ESTs) that could be derived from EPR-1. All ESTs with strong homology to EPR-1/survivin were derived from survivinand could not encode EPR-1. By polymerase chain reaction and Southern blot hybridization, EPR-1 was not detectable in the human or murine genome, but survivin was. Our data suggest that EPR-1 is either highly cell-specific or the published EPR-1 cDNA includes sequences from clones derived from survivin messenger RNA. The means by which factor Xa mediates its cellular effects requires further evaluation.

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The coagulation protease factor Xa induces cellular responses implicated in cardiovascular and inflammatory disease. Effector-cell protease receptor 1 (EPR-1) is a functionally characterized receptor of factor Xa, and the EPR-1complementary DNA (cDNA) was published. Remarkably, the cDNA encoding an inhibitor of apoptosis, survivin, is reportedly identical to that ofEPR-1 except for a few nucleotide differences and its orientation opposite to EPR-1. To isolate the EPR-1cDNA and gene, we surveyed gene databases for expressed sequence tags (ESTs) that could be derived from EPR-1. All ESTs with strong homology to EPR-1/survivin were derived from survivinand could not encode EPR-1. By polymerase chain reaction and Southern blot hybridization, EPR-1 was not detectable in the human or murine genome, but survivin was. Our data suggest that EPR-1 is either highly cell-specific or the published EPR-1 cDNA includes sequences from clones derived from survivin messenger RNA. The means by which factor Xa mediates its cellular effects requires further evaluation.

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Damen, J., AL Mui, P. Hughes, K. Humphries, and G. Krystal. "Erythropoietin-induced tyrosine phosphorylations in a high erythropoietin receptor-expressing lymphoid cell line." Blood 80, no. 8 (October 15, 1992): 1923–32. http://dx.doi.org/10.1182/blood.v80.8.1923.1923.

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Abstract Retroviral gene transfer of the murine erythropoietin receptor (EpR) cDNA into the pro-B-cell line, Ba/F3, was used to generate cells expressing high EpR levels. One of the resulting clones, Ba/F3 clone C5, contained 5 integrated copies of the gene and expressed, at the cell surface, a single affinity class of EpRs at 10 to 15 times the level present on spleen cells from phenylhydrazine-treated mice. Cross- linking studies with clone C5, using 125I-Ep, yielded the same two 105- and 88-Kd major species as that seen with typical erythroid cells. This was distinct from that obtained with EpR-transfected COS cells or L cells, which gave species of 88 and 65 Kd. This suggests that the biologically active EpR complex generated in this Ba/F3 cell line may closely resemble that present in native Ep-responsive erythroid progenitor cells. Tyrosine phosphorylation experiments showed that several proteins in clone C5 cells were rapidly phosphorylated on tyrosine residues in response to Ep, one being the EpR itself. The proportion of cell surface EpRs tyrosine phosphorylated in response to Ep was substantial, reaching a maximum of approximately 10% within 30 minutes of incubation at 37 degrees C. A comparison of Ep- and murine interleukin-3 (mIL-3)-induced tyrosine phosphorylation patterns in clone C5 cells showed that both growth factors stimulated the tyrosine phosphorylation of proteins with molecular weights of 135, 93, 70, and 55 Kd. This could suggest that the Ep and mIL-3 receptors are capable of using the same tyrosine kinase in these cells.

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Retroviral gene transfer of the murine erythropoietin receptor (EpR) cDNA into the pro-B-cell line, Ba/F3, was used to generate cells expressing high EpR levels. One of the resulting clones, Ba/F3 clone C5, contained 5 integrated copies of the gene and expressed, at the cell surface, a single affinity class of EpRs at 10 to 15 times the level present on spleen cells from phenylhydrazine-treated mice. Cross- linking studies with clone C5, using 125I-Ep, yielded the same two 105- and 88-Kd major species as that seen with typical erythroid cells. This was distinct from that obtained with EpR-transfected COS cells or L cells, which gave species of 88 and 65 Kd. This suggests that the biologically active EpR complex generated in this Ba/F3 cell line may closely resemble that present in native Ep-responsive erythroid progenitor cells. Tyrosine phosphorylation experiments showed that several proteins in clone C5 cells were rapidly phosphorylated on tyrosine residues in response to Ep, one being the EpR itself. The proportion of cell surface EpRs tyrosine phosphorylated in response to Ep was substantial, reaching a maximum of approximately 10% within 30 minutes of incubation at 37 degrees C. A comparison of Ep- and murine interleukin-3 (mIL-3)-induced tyrosine phosphorylation patterns in clone C5 cells showed that both growth factors stimulated the tyrosine phosphorylation of proteins with molecular weights of 135, 93, 70, and 55 Kd. This could suggest that the Ep and mIL-3 receptors are capable of using the same tyrosine kinase in these cells.

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Ovcherenko, Sergey S., Olga A. Chinak, Anton V. Chechushkov, Sergey A. Dobrynin, Igor A. Kirilyuk, Olesya A. Krumkacheva, Vladimir A. Richter, and Elena G. Bagryanskaya. "Uptake of Cell-Penetrating Peptide RL2 by Human Lung Cancer Cells: Monitoring by Electron Paramagnetic Resonance and Confocal Laser Scanning Microscopy." Molecules 26, no. 18 (September 7, 2021): 5442. http://dx.doi.org/10.3390/molecules26185442.

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RL2 is a recombinant analogue of a human κ-casein fragment, capable of penetrating cells and inducing apoptosis of cancer cells with no toxicity to normal cells. The exact mechanism of RL2 penetration into cells remains unknown. In this study, we investigated the mechanism of RL2 penetration into human lung cancer A549 cells by a combination of electron paramagnetic resonance (EPR) spectroscopy and confocal laser scanning microscopy. EPR spectra of A549 cells incubated with RL2 (sRL2) spin-labeled by a highly stable 3-carboxy-2,2,5,5-tetraethylpyrrolidine-1-oxyl radical were found to contain three components, with their contributions changing with time. The combined EPR and confocal-microscopy data allowed us to assign these three forms of sRL2 to the spin-labeled protein sticking to the membrane of the cell and endosomes, to the spin-labeled protein in the cell interior, and to spin labeled short peptides formed in the cell because of protein digestion. EPR spectroscopy enabled us to follow the kinetics of transformations between different forms of the spin-labeled protein at a minimal spin concentration (3–16 μM) in the cell. The prospects of applications of spin-labeled cell-penetrating peptides to EPR imaging, DNP, and magnetic resonance imaging are discussed, as is possible research on an intrinsically disordered protein in the cell by pulsed dipolar EPR spectroscopy.

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Bonucci, Alessio, Olivier Ouari, Bruno Guigliarelli, Valérie Belle, and Elisabetta Mileo. "In‐Cell EPR: Progress towards Structural Studies Inside Cells." ChemBioChem 21, no. 4 (September 26, 2019): 451–60. http://dx.doi.org/10.1002/cbic.201900291.

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Dissertations / Theses on the topic "In-Cell EPR"

Pierro, Annalisa. "Protein structural dynamics in bacteria via nitroxide-based SDSL-EPR spectroscopy : from method improvements to in-cell studies." Electronic Thesis or Diss., Aix-Marseille, 2021. http://theses.univ-amu.fr.lama.univ-amu.fr/211116_PIERRO_290xrxu60ryzjfl293g970fjmdnl_TH%20(1).pdf.

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L'étude des biomolécules dans leur environnement natif, la cellule, est l'un des principaux objectifs de la biologie structurale au cours de la dernière décennie. Ainsi, nous assistons à un remarquable développement des approches dites « in-cell », comme la CryoET, FRET (Förster Resonance Energy Tranfer), RMN. Parmi elles, la technique de marquage de spin couplée à la spectroscopie de Résonance Paramagnétique Electronique (SDSL-RPE) présente des caractéristiques intéressantes et avantageuses permettant de sonder la dynamique des protéines à l'intérieur des cellules. En particulier, l’utilisation des sondes de type nitroxyde combine sensibilité élevée et absence de contraintes de taille de la biomolécule d'intérêt avec la capacité d'étudier les transitions structurales et les interactions protéines-protéines à température physiologique. Cependant, même si de nombreux efforts ont été faits pour adapter cette technique à des études structurales dans les cellules, des progrès restent à faire.Dans cette thèse, nous traitons des principales limitations de l’utilisation des nitroxydes dans un contexte cellulaire. Nous nous sommes concentrés sur la stabilité des marqueurs nitroxydes dans des milieux reducteurs et dans la cellule, sur l’incorporation des protéines marquées dans les cellules et la viabilité de ces cellules en vue de mesures par RPE. Grâce aux résultats obtenus dans cette partie méthodologique, nous avons pu étudier la dynamique structurale de deux protéines chaperons (NarJ et UreG) dans des cellules bactériennes. Ces avancées ont permis de comparer les données obtenues in-cell à celles obtenues in vitro ou dans un environnement mimant le milieu cellulaire
The study of biomolecules in their native environment has been one of the main goals of structural biology in the last decade. As a result, we are assisting to a remarkable increase of new "in-cell" approaches, like Cryo-ET, FRET and NMR. Among these approaches, Site-Directed Spin Labeling (SDSL) coupled to Electron Paramagnetic Resonance (EPR) spectroscopy shows competitive and advantageous features to capture protein dynamics inside cells. In particular, nitroxide-based SDSL-EPR combines the advantages of high sensitivity and the lack of size constraints on the biomolecule of interest with the ability to capture protein structural transitions and interactions at physiological temperature. Despite the methodological advancements of the technique that have allowed the community to obtain increasingly relevant results, progresses still need to be done.In this work, the main limitation of nitroxide-based SDSL-EPR has been addressed. In the first time, we focused on the development of delivery methods to introduce the labeled protein in bacterial cells. Next, the stability of nitroxide labels in reducing environments and in-cell has been assessed, monitoring in parallel the viability of the cells during the EPR measurements. Thanks to the results achieved in this methodological part, we were able to study the structural dynamics of two flexible chaperone proteins directly in bacterial cells: NarJ from Escherichia coli and UreG from Sporosarcina pasteurii. Finally, to go further in understanding the impact of the cellular environment on the protein dynamics, the data obtained in cellular context were compared with those obtained in vitro or in a cell-mimicking environment

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Noda, Yohei. "EPR studies on molecular orientation of all-organic paramagnetic liquid crystals in a surface-stabilized liquid crystal cell." 京都大学 (Kyoto University), 2007. http://hdl.handle.net/2433/136793.

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ROSSI, MARTINA. "Long Noncoding RNA “EPR” controls epithelial cell proliferation by coordinating CDKN1A transcription and mRNA decay in response to TGF-β." Doctoral thesis, Università degli studi di Genova, 2019. http://hdl.handle.net/11567/936734.

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Long noncoding RNAs (lncRNAs) are emerging as regulators of fundamental biological processes. Here we report on the functional characterization of an intergenic lncRNA (lincRNA) expressed in epithelial tissues which we termed EPR (Epithelial cell Program Regulator). EPR is rapidly downregulated by TGF-β and its sustained expression largely reshapes the transcriptome, favors the acquisition of epithelial traits, and reduces cell proliferation in cultured mammary gland cells as well as in an animal model of orthotopic transplantation. EPR generates a small peptide that localizes at epithelial cell junctions but the RNA molecule per se accounts for the vast majority of EPR-induced gene expression changes. Mechanistically, EPR interacts with chromatin and regulates Cdkn1a gene expression by affecting both its transcription and mRNA decay through its association with the transcription factor SMAD3 and the mRNA decay promoting factor KHSRP, respectively. We propose that EPR enables epithelial cells to control proliferation by modulating waves of gene expression in response to stimuli.

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Udyaningsih-Freisleben, Seruni Kusuma. "XAS and RR Structural Analysis of Hemoglobin and EPR Spectroscopic Labelling of Red Blood Cell Membranes Isolated from Thalassemia Patients in Jakarta, Indonesia." Thesis, The University of Sydney, 2003. https://hdl.handle.net/2123/27995.

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A study on thalassemia intermedia and major patients in Jakarta was initiated to obtain a comprehensive picture of metabolic dysregulation, iron overload, oxidative stress, and cell damage. Data were collected from a group of 10 transfusion-dependent patients in an age range of 11-25 years and another group of 5 frequently transfused (for at least 15 years) patients aged 17-30 years. A third group comprises 5 patients (aged 7 to 14 years) who had not yet obtained transfusions. The 10 controls were voluntary students without diagnosis or clinical signs of thalassemia up to 30 years of age. The study was approved by the Ethical Clearance Board of the Medical Faculty and all blood samples from controls and patients were obtained on fully informed consent. Levels of antioxidants (vitamins A, C, E and B-carotene) and reactive thiols are considerably decreased in transfused patients, whereas signs of iron overload and cell damage are increased (serum iron, ferritin, transferrin saturation, SGOT, SGPT, y-GT, bilirubin).

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Fernandez, Daniel. "Cell States and Cell Fate: Statistical and Computational Models in (Epi)Genomics." Thesis, Harvard University, 2015. http://nrs.harvard.edu/urn-3:HUL.InstRepos:14226043.

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This dissertation develops and applies several statistical and computational methods to the analysis of Next Generation Sequencing (NGS) data in order to gain a better understanding of our biology. In the rest of the chapter we introduce key concepts in molecular biology, and recent technological developments that help us better understand this complex science, which, in turn, provide the foundation and motivation for the subsequent chapters. In the second chapter we present the problem of estimating gene/isoform expression at the allelic level, and different models to solve this problem. First, we describe the observed data and the computational workflow to process the data. Next, we propose frequentist and bayesian models motivated by the central dogma of molecular biology and the data generating process (DGP) for RNA-Seq. We develop EM and Gibbs sampling approaches to estimate gene and transcript-specic expression from our proposed models. Finally, we present the performance of our models in simulations and we end with the analysis of experimental RNA-Seq data at the allelic level. In the third chapter we present our paired factorial experimental design to study parentally biased gene/isoform expression in the mouse cerebellum, and dynamic changes of this pattern between young and adult stages of cerebellar development. We present a bayesian variable selection model to estimate the difference in expression between the paternal and maternal genes, while incorporating relevant factors and its interactions into the model. Next, we apply our model to our experimental data, and further on we validate our predictions using pyrosequencing follow-up experiments. We subsequently applied our model to the pyrosequencing data across multiple brain regions. Our method, combined with the validation experiments, allowed us to find novel imprinted genes, and investigate, for the first time, imprinting dynamics across brain regions and across development. In the fourth chapter we move from the controlled-experiments in mouse isogenic lines to the highly variant world of human genetics in observational studies. In this chapter we introduce a Bayesian Regression Allelic Imbalance Model, BRAIM, that estimates the imbalance coming from two major sources: cis-regulation and imprinting. We model the cis-effect as an additive effect for the heterozygous group and we model the parent-of-origin detect with a latent variable that indicates to which parent a given allele belongs. Next, we show the performance of the model under simulation scenarios, and finally we apply the model to several experiments across multiple tissues and multiple individuals. In the fifth chapter we characterize the transcriptional regulation and gene expression of in-vitro Embryonic Stem Cells (ESCs), and two-related in-vivo cells; the Inner Cell Mass (ICM) tissue, and the embryonic tissue at day 6.5. Our objective is two fold. First we would like to understand the differences in gene expression between the ESCs and their in-vivo counterpart from where these cells were derived (ICM). Second, we want to characterize the active transcriptional regulatory regions using several histone modifications and to connect such regulatory activity with gene expression. In this chapter we used several statistical and computational methods to analyze and visualize the data, and it provides a good showcase of how combining several methods of analysis we can delve into interesting developmental biology.

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Hu, Zhengqing. "Investigating a cell replacement therapy in the inner ear /." Stockholm, 2004. http://diss.kib.ki.se/2005/91-7140-170-9/.

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Gregory, L. G. L. "Eph-ephrin signalling in cell sorting and directional migration." Thesis, University College London (University of London), 2011. http://discovery.ucl.ac.uk/1318081/.

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An important problem in developmental biology is to understand how precise patterns of cell types are maintained during development. Eph receptor tyrosine kinases and ephrins have key roles in stabilising these patterns of cell organisation and segregation during development and can restrict the movement of cells by promoting cell repulsion. Previous work by Alexei Poliakov in the Wilkinson lab has shown that Eph-ephrin signalling leads to directional persistence of migration, and modelling suggests that this can contribute to cell segregation. In order to test experimentally the contribution of directional persistence in cell segregation, I have used and developed in vitro assays to dissect the roles of EphB2-ephrinB1 signalling in cell segregation, boundary sharpening and directional persistence. In these assays, stable HEK293 cell lines expressing EphB2 or ephrinB1 are mixed in cell culture and this leads to segregation of the two cell populations. Plating these cells either side of a removable barrier and allowing migration of cells towards each other leads to the formation of a sharp boundary on interaction. Analysis of cell behaviour shows EphB2 cells to move more persistently after interaction with ephrinB1 cells. To analyse how EphB2-ephrinB1 interactions lead to directional persistence of migration, my studies have focussed on the role of components potentially involved in directional persistence that act downstream of EphB2-ephrinB1 signalling, including the planar cell polarity (PCP) pathway (Dishevelled and Daam1) and core polarity components such as the PAR proteins (PAR-3 and PAR-6B). The PCP and PAR components were all found to have roles in cell segregation, as siRNA-mediated knockdown of each of these components disrupted EphB2-ephrinB1 mediated cell segregation and boundary sharpening. However, cell behaviour studies showed that only Dishevelled and PAR-6B have roles in EphB2-ephrinB1 mediated directional persistence, whilst Daam1 knockdown has no effect on the migratory response of cells. PAR-3 knockdown affects the basal ability of cells to migrate, potentially due to its role in establishing front-rear polarity. Taken together, these findings can be explained by a model in which Dishevelled and PAR-6B have a role in EphB2-ephrinB1 mediated directional persistence required for cell segregation and boundary sharpening. I propose that Daam1 may function in the contact inhibition of locomotion between cells also required for segregation.

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Li, Lin. "Hair cell loss and repair processes in mammalian vestibular sensory epithelia." Thesis, University College London (University of London), 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.266075.

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Omodho, Becky. "Early growth response gene (Egr) 2 and 3 control inflammatory responses of tolerant T cells." Thesis, Brunel University, 2016. http://bura.brunel.ac.uk/handle/2438/13516.

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This study investigated the role of tolerance induction in an inflammatory setting in regard to the early growth response genes Egr2 and Egr3. T cells robustly respond to pathogenic antigens during infection, but are tolerant to stimulation by self-antigens. The intrinsic mechanisms for self-tolerance in the periphery are still not clear. Egr2 and 3 are induced in tolerant T cells in response to antigen stimulation by NFAT-medicated tolerant signalling; however, their function in tolerant T cells is still unknown. The study demonstrated that Egr2 and 3, induced in tolerant T cells, are not directly involved in defective proliferation and IL-2 production, the hallmarks of T cell tolerance. However, they are essential for preventing inflammatory response of tolerant T cells. In the absence of Egr2 and 3, tolerant T cells show impaired proliferation and production of IL-2, but produce high levels of IFN-γ, a key inflammatory cytokine. This phenotype resembles CD4 T cells from autoimmune diseases such as lupus which show poor proliferative response, but hyper-inflammation. Our study demonstrated, for the first time, a distinctive mechanism to control inflammation from proliferative tolerance regulated by Egr2 and 3, which may be an important mechanism for the control of autoimmune diseases.

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Gallagher, Ewen. "The regulation of the egr-1 promoter in B cell lines." Thesis, University of Nottingham, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.341970.

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Books on the topic "In-Cell EPR"

C. Lu, C. Han*, W. Yang, F. Ye, F. He, S. Wei, H. Liu, L. Li, and H. Xu. Glucose and insulin regulate cell proliferation and glucose transporter (GLUT) synthesis in the liver of overfed geese. Verlag Eugen Ulmer, 2021. http://dx.doi.org/10.1399/eps.2021.334.

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S.M. Hosseini, M. Bashtani, H. Nazarizadeh, and M. Manafi. Effects of diet form and particle size on performance, digestive tract development and intestinal mast cell numbers in young broiler chicks. Verlag Eugen Ulmer, 2016. http://dx.doi.org/10.1399/eps.2016.161.

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Kruger, Robert Paul. A novel receptor-like tyrosine phosphatase expressed in the avian inner ear: Implications for hair cell regeneration. 2000.

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Provan, Drew, Trevor Baglin, Inderjeet Dokal, and Johannes de Vos. Clinical approach. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199683307.003.0001.

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History taking in patients with haematological disease - Physical examination - Splenomegaly - Lymphadenopathy - Unexplained anaemia - Patient with elevated haemoglobin - Elevated white blood cell (WBC) count - Reduced WBC count - Elevated platelet count - Reduced platelet count - Easy bruising - Recurrent thromboembolism - Pathological fracture - Raised ESR - Serum or urine paraprotein - Anaemia in pregnancy - Thrombocytopenia in pregnancy - Prolonged bleeding after surgery - Positive sickle test (HbS solubility test)

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Provan, Drew, Trevor Baglin, Inderjeet Dokal, Johannes de Vos, and Hassan Al-Sader. Clinical approach. Oxford University Press, 2018. http://dx.doi.org/10.1093/med/9780199683307.003.0001_update_001.

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Wackerhage, Henning, Jonathon Smith, and Darren Wisniewski. Molecular exercise physiology. Edited by Neil Armstrong and Willem van Mechelen. Oxford University Press, 2017. http://dx.doi.org/10.1093/med/9780198757672.003.0031.

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Molecular exercise physiology is the study of exercise physiology using molecular biology methods. The development of differentiated cell types is regulated by transcription factors like the muscle-making MyoD that specifies cell type, while others regulate the development of muscle, tendons, and bones. Maternal nutrition and exercise commonly affect embryonic development through epigenetic mechanisms. Adaptation to exercise involves sensor proteins detecting exercise-related signals, the processing of signals by signalling proteins and networks, and the regulation of the actual adaptations by effector proteins. Many sport- and exercise-related traits depend on both common and rare DNA sequence variations, including the muscle mass-increasing myostatin (GDF8) loss-of-function and the haematocrit-increasing EPOR gain-of-function mutations. Additionally, common DNA sequence variations contribute to the inherited variability of development, body height, strength, and endurance. Finally, in addition to ethical concerns, current genetic performance tests only explain a fraction of the variation of sport and exercise-related traits.

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Kapoor, Reena. Crisis assessment and management. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199360574.003.0025.

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Crisis calls are a common occurrence in correctional settings. Psychiatrists are often called upon to triage and manage such events. Requests for urgent psychiatric evaluations can come from many sources, including security staff, non-psychiatric physicians, mental health staff, courts, attorneys, and family members. Psychiatrists responding to these requests for evaluation may feel tremendous pressure to reach a conclusion that is consistent with the opinions of the requesting party. However, maintaining an independent and therapeutic stance when conducting crisis evaluations is crucial. Some aspects of psychiatric evaluations in crisis situations are unique to the correctional environment: evaluations at cell-side, video recording, and leadership by security staff rather than medical professionals. Nonetheless, correctional psychiatrists should be guided by the same principles of medical ethics that apply to patient care in the community, placing the patient’s well-being above all other concerns. They should strive, when possible, to conduct a thorough assessment in a confidential setting. In considering how best to resolve the crisis and care for the patient, they should err on the side of caution and recommend placement in a safe and therapeutic setting, at least until a multidisciplinary team can consider other options. Finally, they should document the encounter carefully, articulating the rationale for the chosen course of action. This chapter reviews the pragmatics of evaluating and managing many common correctional events that lead to mental health crisis calls and discusses the range of concerns, the typical practices and procedures used in correctional settings, and the types of interventions that are best used.

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Nakamura, Tomohiro, and Stuart A. Lipton. Neurodegenerative Diseases as Protein Misfolding Disorders. Oxford University Press, 2016. http://dx.doi.org/10.1093/med/9780190233563.003.0002.

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Neurodegenerative diseases (NDDs) often represent disorders of protein folding. Rather than large aggregates, recent evidence suggests that soluble oligomers of misfolded proteins are the most neurotoxic species. Emerging evidence points to small, soluble oligomers of misfolded proteins as the cause of synaptic dysfunction and loss, the major pathological correlate to disease progression in many NDDs including Alzheimer’s disease. The protein quality control machinery of the cell, which includes molecular chaperones as found in the endoplasmic reticulum (ER), the ubiquitin-proteasome system (UPS), and various forms of autophagy, can counterbalance the accumulation of misfolded proteins to some extent. Their ability to eliminate the neurotoxic effects of misfolded proteins, however, declines with age. A plausible explanation for the age-dependent deterioration of the quality control machinery involves compromise of these systems by excessive generation of reactive oxygen species (ROS), such as superoxide anion (O2-), and reactive nitrogen species (RNS), such as nitric oxide (NO). The resulting redox stress contributes to the accumulation of misfolded proteins. Here, we focus on aberrantly increased generation of NO-related species since this process appears to accelerate the manifestation of key neuropathological features, including protein misfolding. We review the chemical mechanisms of posttranslational modification by RNS such as protein S-nitrosylation of critical cysteine thiol groups and nitration of tyrosine residues, showing how they contribute to the pathogenesis of NDDs.

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Book chapters on the topic "In-Cell EPR"

Bieber, Anna, and Malte Drescher. "Chapter 10. In-cell EPR." In New Developments in NMR, 152–69. Cambridge: Royal Society of Chemistry, 2019. http://dx.doi.org/10.1039/9781788013079-00152.

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Weickert, Sabrina, Julia Cattani, and Malte Drescher. "Intrinsically disordered proteins (IDPs) studied by EPR and in-cell EPR." In Electron Paramagnetic Resonance, 1–37. Cambridge: Royal Society of Chemistry, 2018. http://dx.doi.org/10.1039/9781788013888-00001.

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Ilangovan, Govindasamy, Haiquan Li, Jay L. Zweier, and Periannan Kuppusamy. "In vivo measurement of tumor redox environment using EPR spectroscopy." In Oxygen/Nitrogen Radicals: Cell Injury and Disease, 393–98. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/978-1-4615-1087-1_44.

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Kita, Tomoko. "Hair Cell Regeneration in the Avian." In Regenerative Medicine for the Inner Ear, 181–88. Tokyo: Springer Japan, 2014. http://dx.doi.org/10.1007/978-4-431-54862-1_19.

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Khan, Nadeem, and Harold Swartz. "Measurements in vivo of parameters pertinent to ROS/RNS using EPR spectroscopy." In Oxygen/Nitrogen Radicals: Cell Injury and Disease, 341–57. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/978-1-4615-1087-1_39.

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Hinrichsen, Klaus. "Early Development of the External Ear." In Advances in Anatomy Embryology and Cell Biology, 66. Berlin, Heidelberg: Springer Berlin Heidelberg, 1985. http://dx.doi.org/10.1007/978-3-642-70754-4_10.

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Székely, George, and Clara Matesz. "The Muscles of the Middle Ear." In Advances in Anatomy Embryology and Cell Biology, 39–42. Berlin, Heidelberg: Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-77938-1_7.

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Murayama, Toshinori, Oren M. Tepper, and Takayuki Asahara. "Murine Bone Marrow Transplantation Models that Enable the Study of EPC Recruitment." In Methods in Endothelial Cell Biology, 179–85. Berlin, Heidelberg: Springer Berlin Heidelberg, 2004. http://dx.doi.org/10.1007/978-3-642-18725-4_17.

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Rubel, Edwin W., Elizabeth C. Oesterle, and Pedro Weisleder. "Hair Cell Regeneration in the Avian Inner Ear." In Ciba Foundation Symposium 160 - Regeneration of Vertebrate Sensory Receptor Cells, 77–102. Chichester, UK: John Wiley & Sons, Ltd., 2007. http://dx.doi.org/10.1002/9780470514122.ch5.

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Sun, Hong, Aji Huang, Hong Liu, and Shousong Cao. "Gene Therapy for the Inner Ear: Progress and Prospects." In Emerging Trends in Cell and Gene Therapy, 595–623. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-417-3_24.

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Conference papers on the topic "In-Cell EPR"

Lubart, Rachel, R. Lavie, Harry Friedmann, M. Sinyakov, Asher Shainberg, Haim Breitbart, and Nili Grossman. "UVA and visible light-induced reactive oxygen species (ROS) formation in cell cultures: an electron paramagnetic resonance (EPR) study." In EOS/SPIE European Biomedical Optics Week, edited by Tiina I. Karu and Rachel Lubart. SPIE, 2000. http://dx.doi.org/10.1117/12.405915.

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Argitis, P., A. C. Cef alas, Z. Kollia, E. Sarantopoulou, T. W. Ford, A. D. Stead, A. Marranca, C. N. Danson, J. Knott, and D. Neely. "Fast, Chemically Amplified Epoxy Novolac Photoresist for Soft X- Ray Contact Microscopy of Living Biological Species." In The European Conference on Lasers and Electro-Optics. Washington, D.C.: Optica Publishing Group, 1998. http://dx.doi.org/10.1364/cleo_europe.1998.cmd6.

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Soft X-ray contact microscopy (SXCM), has many applications in both life and material sciences. In the case of life sciences SXCM enables the study of the ultrastructure of living hydrated specimens, without the need of dehydration or other chemical pretreatment, by using suitable pulsed X-ray sources such as laser plasmas [1,2]. The interest in using soft X-rays, in the so called “water window” (2.3-4.4nm), is based on the low attenuation at these wavelengths caused by water, as compared to the attenuation caused by organic matter. Therefore, good contrast masking of the incident radiation is provided. The successful imaging of biological specimen, requires the development of sensitive photoresist materials for image recording; these should have capabilities of high resolution lithography and an extended grayscale. Up to now, the only known photoresist used successfully in SXCM has been polymethyl methacrylate (PMMA). This is a high resolution photoresist when exposed to e-beam or X-ray radiation, with contrast suitable for gray scale recording ; nevertheless, it is a relatively slow photoresist and, therefore, requires a very large fluence of X-rays for imaging. The work reported here was carried out using the Vulcan Nd:glass laser facility at the Rutherford Appleton Laboratory UK, whose rod chain output can deliver more than 11 J at 1064nm. This can be delivered on a Yttrium target as an X- ray source. A very sensitive e -- beam photoresist, used for the first time in SXCM, enabled the biological imaging with the specific source in single pulse experiments in the water window spectral range. This photoresist is an epoxy novolac based chemically amplified photoresist (EPR) which has been proven capable of resolving sub tenth micron features. Initial experiments to compare the sensitivity of PMMA and EPR were done in the absence of a biological specimen. The first image for EPR is obtained at 300mJ laser pulse energy. PMMA used as a reference gave a first image of 40nm depth difference between exposed and unexposed areas, as measured with a Dektak profilometer, at 4.6 J laser pulse energy and an image of 70 nm at 10.6 J. Thus on the basis of the calculations of X-ray flux produced at different laser energies, the minimum flux for image production with PMMA is 2.5mJ.cm-2 and the corresponding value for EPR is only ~ 0.07 mJ.cm-2, giving a difference of two orders of magnitude approximately, for the two materials. In biological imaging experiments the living specimens were cells of the motile green alga, Chlamydomonas, which were placed in a droplet of medium. In the experiments with biological specimens no image was obtained with PMMA as a recording material, even with the higher pulse energy available and careful adjustment of water layer thickness in order to be exactly the size of cell diameter. In the contrary, with the EPR resist biological imaging was possible. Images of Chlamydomonas cells were successfully obtained. These images clearly show the cell body and the flagella, suggest a lateral resolution considerably better than 300nm.

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Kwak, Bongseop, Kinam Park, and Bumsoo Han. "Tumor-on-Chip: Simulation of Complex Transport Around Tumor." In ASME 2013 2nd Global Congress on NanoEngineering for Medicine and Biology. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/nemb2013-93314.

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Nanoparticles (NP) offer great potential as drug carriers for targeted delivery to tumor by increasing the delivery efficacy and reducing non-specific accumulation at non-targeted sites [1]. Despite these promising early outcomes [2], the NP delivery to target tumor site is still significantly limited due to complex in vivo transport barriers [3–5]. In order to improve the in vivo delivery efficacy, the NPs should be designed considering all these complex transport barriers beyond currently used enhanced permeation and retention (EPR) effect [6]. However, testing of NP delivery are primarily based on simple 2D or 3D in vitro cell cultures or animal models. However, these static 2D or 3D tumor models oversimplify the actual in vivo tumor environment including the absence of tissue-tissue interactions such as blood-endothelium, endothelium-intersititum, high interstitial fluid pressure, and interstitium-lymphatics [2, 3]. The animal models can provide the testbed with these tissue-tissue interactions, but it is very difficult to establish quantitative understanding of the NP transport at these tissue-tissue interfaces. To address these challenges and bridge the in vitro static models with the animal models, here we developed a 3D multi-layered microfluidic system that mimics the tissue-tissue interactions at tumor microenvironment is developed. The system is then used to investigate the transvascular and interstitial transport of NPs in tumor.

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Aso, Hiromichi, Satoru Ito, Akemi Mori, Masataka Morioka, Nobukazu Suganuma, Masashi Kondo, and Yoshinori Hasegawa. "Prostaglandin E2 Inhibits Platelet-Derived Growth Factor-Induced Cell Migration Via EP2 And EP4 Receptors In Human Airway Smooth Muscle Cells." In American Thoracic Society 2012 International Conference, May 18-23, 2012 • San Francisco, California. American Thoracic Society, 2012. http://dx.doi.org/10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a2420.

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Warnecke, A., M. Steffens, J. Schulze, K. Willenborg, N. Prenzler, and T. Lenarz. "Stem cell survival in inner ear fluids: implications for cell transplantation into the endolymphatic sac." In Abstract- und Posterband – 89. Jahresversammlung der Deutschen Gesellschaft für HNO-Heilkunde, Kopf- und Hals-Chirurgie e.V., Bonn – Forschung heute – Zukunft morgen. Georg Thieme Verlag KG, 2018. http://dx.doi.org/10.1055/s-0038-1641063.

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Spector, Alexander A., William E. Brownell, and Aleksander S. Popel. "An Elastic Model of a Composite Cell Membrane: Interpretation and Design of Experiments." In ASME 1996 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 1996. http://dx.doi.org/10.1115/imece1996-1135.

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Abstract Background. We study the elastic properties of the outer hair cell. The outer hair cell is a receptor cell located in the inner ear and recently considered as the major active component in the hearing process. The lateral wall of this cell is able to transmit the changes of the membrane potential into the changes of the length of the cell (Brownell et al., 1985). The outer hair cell is an elongated cylinder filled with a fluid and embedded into a surrounding structure of deformable elements. Through the electromechanical transduction, outer hair cells exert forces on the basilar membrane, the major vibrating element in the cochlea, and are believed to affect the tuning properties of the ear. Therefore, the elastic properties of the outer hair cell are crucial for the analysis of the role of the active element and for understanding the hearing process in general.

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Ospeck, Mark. "Noise in the Outer Hair Cell and Gain Control in the Ear." In UNSOLVED PROBLEMS OF NOISE AND FLUCTUATIONS: UPoN 2002: Third International Conference on Unsolved Problems of Noise and Fluctuations in Physics, Biology, and High Technology. AIP, 2003. http://dx.doi.org/10.1063/1.1584882.

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Sonderskov, Simon Heindorf, Dean Rasmussen, Jakob Ilsoe, Daniel Blom-Hansen, and Stig Munk-Nielsen. "Detecting Performance Outliers in Fuel Cell Backup Power Systems." In 2019 21st European Conference on Power Electronics and Applications (EPE '19 ECCE Europe). IEEE, 2019. http://dx.doi.org/10.23919/epe.2019.8915057.

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Rusu, Florin Andrei, Alina Georgiana Baciu, and Gheorghe Livint. "Applicability of Fuel Cell in Electric Vehicles." In 2018 International Conference and Exposition on Electrical And Power Engineering (EPE). IEEE, 2018. http://dx.doi.org/10.1109/icepe.2018.8559669.

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Chitakudige, Ramachandra, Sarat Kumar Dash, and A. M. Khan. "Deprocessing Methodologies for Detection of IBC and Cell-to-Cell Shorts in Submicron DRAM." In ISTFA 2012. ASM International, 2012. http://dx.doi.org/10.31399/asm.cp.istfa2012p0383.

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Abstract Detection of both Insufficient Buried Contact (IBC) and cell-to-cell short defects is quite a challenging task for failure analysis in submicron Dynamic Random Access Memory (DRAM) devices. A combination of a well-controlled wet etch and high selectivity poly silicon etch is a key requirement in the deprocessing of DRAM for detection of these types of failures. High selectivity poly silicon etch methods have been reported using complicated system such as ECR (Electron Cyclotron Resonance) Plasma system. The fact that these systems use hazardous gases like Cl2, HBr, and SF6 motivates the search for safer alternative deprocessing chemistries. The present work describes high selectivity poly silicon etch using simple Reactive Ion Etch (RIE) plasma system using less hazardous gases such as CF4, O2 etc. A combination of controlled wet etch and high selectivity poly silicon etch have been used to detect both IBC and cell-to-cell shorts in submicron DRAMs.

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Reports on the topic "In-Cell EPR"

Epel, Bernard, and Roger Beachy. Mechanisms of intra- and intercellular targeting and movement of tobacco mosaic virus. United States Department of Agriculture, November 2005. http://dx.doi.org/10.32747/2005.7695874.bard.

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To cause disease, plant viruses must replicate and spread locally and systemically within the host. Cell-to-cell virus spread is mediated by virus-encoded movement proteins (MPs), which modify the structure and function of plasmodesmata (Pd), trans-wall co-axial membranous tunnels that interconnect the cytoplasm of neighboring cells. Tobacco mosaic virus (TMV) employ a single MP for cell- cell spread and for which CP is not required. The PIs, Beachy (USA) and Epel (Israel) and co-workers, developed new tools and approaches for study of the mechanism of spread of TMV that lead to a partial identification and molecular characterization of the cellular machinery involved in the trafficking process. Original research objectives: Based on our data and those of others, we proposed a working model of plant viral spread. Our model stated that MPᵀᴹⱽ, an integral ER membrane protein with its C-terminus exposed to the cytoplasm (Reichel and Beachy, 1998), alters the Pd SEL, causes the Pd cytoplasmic annulus to dilate (Wolf et al., 1989), allowing ER to glide through Pd and that this gliding is cytoskeleton mediated. The model claimed that in absence of MP, the ER in Pd (the desmotubule) is stationary, i.e. does not move through the Pd. Based on this model we designed a series of experiments to test the following questions: -Does MP potentiate ER movement through the Pd? - In the presence of MP, is there communication between adjacent cells via ER lumen? -Does MP potentiate the movement of cytoskeletal elements cell to cell? -Is MP required for cell-to-cell movement of ER membranes between cells in sink tissue? -Is the binding in situ of MP to RNA specific to vRNA sequences or is it nonspecific as measured in vitro? And if specific: -What sequences of RNA are involved in binding to MP? And finally, what host proteins are associated with MP during intracellular targeting to various subcellular targets and what if any post-translational modifications occur to MP, other than phosphorylation (Kawakami et al., 1999)? Major conclusions, solutions and achievements. A new quantitative tool was developed to measure the "coefficient of conductivity" of Pd to cytoplasmic soluble proteins. Employing this tool, we measured changes in Pd conductivity in epidermal cells of sink and source leaves of wild-type and transgenic Nicotiana benthamiana (N. benthamiana) plants expressing MPᵀᴹⱽ incubated both in dark and light and at 16 and 25 ᵒC (Liarzi and Epel, 2005 (appendix 1). To test our model we measured the effect of the presence of MP on cell-to-cell spread of a cytoplasmic fluorescent probe, of two ER intrinsic membrane protein-probes and two ER lumen protein-probes fused to GFP. The effect of a mutant virus that is incapable of cell-to-cell spread on the spread of these probes was also determined. Our data shows that MP reduces SEL for cytoplasmic molecules, dilates the desmotubule allowing cell-cell diffusion of proteins via the desmotubule lumen and reduces the rate of spread of the ER membrane probes. Replicase was shown to enhance cell-cell spread. The data are not in support of the proposed model and have led us to propose a new model for virus cell-cell spread: this model proposes that MP, an integral ER membrane protein, forms a MP:vRNAER complex and that this ER-membrane complex diffuses in the lipid milieu of the ER into the desmotubule (the ER within the Pd), and spreads cell to cell by simple diffusion in the ER/desmotubule membrane; the driving force for spread is the chemical potential gradient between an infected cell and contingent non-infected neighbors. Our data also suggests that the virus replicase has a function in altering the Pd conductivity. Transgenic plant lines that express the MP gene of the Cg tobamovirus fused to YFP under the control the ecdysone receptor and methoxyfenocide ligand were generated by the Beachy group and the expression pattern and the timing and targeting patterns were determined. A vector expressing this MPs was also developed for use by the Epel lab . The transgenic lines are being used to identify and isolate host genes that are required for cell-to-cell movement of TMV/tobamoviruses. This line is now being grown and to be employed in proteomic studies which will commence November 2005. T-DNA insertion mutagenesis is being developed to identify and isolate host genes required for cell-to-cell movement of TMV.

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Kamrani, Sonia. Cell Cycle Regulatory Roles of ER-Alpha in Breast Cancer. Fort Belvoir, VA: Defense Technical Information Center, March 2007. http://dx.doi.org/10.21236/ada471166.

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Kalashnikova, Ekaterina. Role of a Novel ER Coactivator in Control of Cell Proliferation and Tamoxifen Resistance. Fort Belvoir, VA: Defense Technical Information Center, October 2009. http://dx.doi.org/10.21236/ada520035.

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Sharon, Amir, and Maor Bar-Peled. Identification of new glycan metabolic pathways in the fungal pathogen Botrytis cinerea and their role in fungus-plant interactions. United States Department of Agriculture, 2012. http://dx.doi.org/10.32747/2012.7597916.bard.

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The involvement of glycans in microbial adherence, recognition and signaling is often a critical determinant of pathogenesis. Although the major glycan components of fungal cell walls have been identified there is limited information available on its ‘minor sugar components’ and how these change during different stages of fungal development. Our aim was to define the role of Rhacontaining-glycans in the gray mold disease caused by the necrotrophic fungus B. cinerea. The research was built on the discovery of two genes, Bcdhand bcer, that are involved in formation of UDP-KDG and UDP-Rha, two UDP- sugars that may serve as donors for the synthesis of cell surface glycans. Objectives of the proposed research included: 1) To determine the function of B. cinereaBcDh and BcEr in glycan biosynthesis and in pathogenesis, 2) To determine the expression pattern of BcDH and BcERand cellular localization of their encoded proteins, 3) Characterize the structure and distribution of Rha- containing glycans, 4) Characterization of the UDP-sugar enzymes and potential of GTs involved in glycanrhamnosylation. To address these objectives we generated a series of B. cinereamutants with modifications in the bchdhand bcergenes and the phenotype and sugar metabolism in the resulting strains were characterized. Analysis of sugar metabolites showed that changes in the genes caused changes in primary and secondary sugars, including abolishment of rhamnose, however abolishment of rhamnose synthesis did not cause changes in the fungal phenotype. In contrast, we found that deletion of the second gene, bcer, leads to accumulation of the intermediate sugar – UDP- KDG, and that such mutants suffer from a range of defects including reduced virulence. Further analyses confirmed that UDP-KDG is toxic to the fungus. Studies on mode of action suggested that UDP-KDG might affect integrity of the fungal cell wall, possibly by inhibiting UDP-sugars metabolic enzymes. Our results confirm that bcdhand bcerrepresent a single pathway of rhamnose synthesis in B. cinerea, that rhamnose does not affect in vitro development or virulence of the fungus. We also concluded that UDP-KDG is toxic to B. cinereaand hence UDP-KDG or compounds that inhibit Er enzymes and lead to accumulation of UDP-KDG might have antifungal activity. This toxicity is likely the case with other fungi, this became apparent in a collaborative work with Prof. Bart Thomma of Wageningen University, NETHERLANDS . We have shown the deletion of ER mutant in Verticillium dahlia gave plants resistance to the fungal infection.

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Tran, Emily, Jasmine J. Park, Nandini N. Kulkarni, and Vinay S. Gundlapalli. Left Facial Primary Leiomyosarcoma Misdiagnosed as Atypical Fibroxanthoma and Immunochemical Markers Relevant to Diagnosis: A Case Report. Science Repository, February 2024. http://dx.doi.org/10.31487/j.ajscr.2023.04.03.

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Soft tissue sarcomas are relatively rare neoplasms of mesenchymal origin that generally make up less than 2% of all adult malignant neoplasms. Atypical fibroxanthoma is a benign soft tissue tumor often confused with malignant variants of similar tumors such as leiomyosarcoma due to similar staining markers and cell morphology. We report a case of a 70-year-old caucasian male who initially presented with a 2 cm exophytic left facial lesion that was misdiagnosed as atypical fibroxanthoma upon biopsy. The patient underwent a wide local excision of the growing 11 cm mass and immediate reconstruction with a cervicofacial flap and full thickness skin graft. Pathological analysis of the specimen revealed the final diagnosis as confirmed primary leiomyosarcoma. Both the patient’s biopsy report and the surgical pathology report revealed similar negative findings (desmin, cytokeratin AE1/AE3, p63, SOX10) as well as similar positive findings (smooth muscle actin and CD68). Critical distinctions that led to a change in diagnosis from atypical fibroxanthoma to leiomyosarcoma emerged during the final pathological analysis, which revealed more widespread positive staining for smooth muscle actin and muscle-specific actin throughout the surgical specimen along with detailed cell and nucleus morphology of atypical spindle cells in the dermis and subcutis. This valuable information was not available during the initial biopsy when the lesion was smaller. It is possible that earlier diagnosis of primary leiomyosarcoma could have resulted in advanced pre-operative treatment and excision of the facial lesion, preventing involvement of surrounding areas such as the patient’s left eye, ear, and facial nerve.

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Or, Dani, Shmulik Friedman, and Jeanette Norton. Physical processes affecting microbial habitats and activity in unsaturated agricultural soils. United States Department of Agriculture, October 2002. http://dx.doi.org/10.32747/2002.7587239.bard.

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experimental methods for quantifying effects of water content and other dynamic environmental factors on bacterial growth in partially-saturated soils. Towards this end we reviewed critically the relevant scientific literature and performed theoretical and experimental studies of bacterial growth and activity in modeled, idealized and real unsaturated soils. The natural wetting-drying cycles common to agricultural soils affect water content and liquid organization resulting in fragmentation of aquatic habitats and limit hydraulic connections. Consequently, substrate diffusion pathways to soil microbial communities become limiting and reduce nutrient fluxes, microbial growth, and mobility. Key elements that govern the extent and manifestation of such ubiquitous interactions include characteristics of diffusion pathways and pore space, the timing, duration, and extent of environmental perturbations, the nature of microbiological adjustments (short-term and longterm), and spatial distribution and properties of EPS clusters (microcolonies). Of these key elements we have chosen to focus on a manageable subset namely on modeling microbial growth and coexistence on simple rough surfaces, and experiments on bacterial growth in variably saturated sand samples and columns. Our extensive review paper providing a definitive “snap-shot” of present scientific understanding of microbial behavior in unsaturated soils revealed a lack of modeling tools that are essential for enhanced predictability of microbial processes in soils. We therefore embarked on two pronged approach of development of simple microbial growth models based on diffusion-reaction principles to incorporate key controls for microbial activity in soils such as diffusion coefficients and temporal variations in soil water content (and related substrate diffusion rates), and development of new methodologies in support of experiments on microbial growth in simple and observable porous media under controlled water status conditions. Experimental efforts led to a series of microbial growth experiments in granular media under variable saturation and ambient conditions, and introduction of atomic force microscopy (AFM) and confocal scanning laser microscopy (CSLM) to study cell size, morphology and multi-cell arrangement at a high resolution from growth experiments in various porous media. The modeling efforts elucidated important links between unsaturated conditions and microbial coexistence which is believed to support the unparallel diversity found in soils. We examined the role of spatial and temporal variation in hydration conditions (such as exist in agricultural soils) on local growth rates and on interactions between two competing microbial species. Interestingly, the complexity of soil spaces and aquatic niches are necessary for supporting a rich microbial diversity and the wide array of microbial functions in unsaturated soils. This project supported collaboration between soil physicists and soil microbiologist that is absolutely essential for making progress in both disciplines. It provided a few basic tools (models, parameterization) for guiding future experiments and for gathering key information necessary for prediction of biological processes in agricultural soils. The project sparked a series of ongoing studies (at DTU and EPFL and in the ARO) into effects of soil hydration dynamics on microbial survival strategy under short term and prolonged desiccation (important for general scientific and agricultural applications).

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Philosoph-Hadas, Sonia, Peter B. Kaufman, Shimon Meir, and Abraham H. Halevy. Inhibition of the Gravitropic Shoot Bending in Stored Cut Flowers Through Control of Their Graviperception: Involvement of the Cytoskeleton and Cytosolic Calcium. United States Department of Agriculture, December 2005. http://dx.doi.org/10.32747/2005.7586533.bard.

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Original objectives: The basic goal of the present project was to study the mechanism involved in shoot graviperception and early transduction, in order to determine the sequence of events operating in this process. This will enable to control the entire process of gravity-induced differential growth without affecting vertical growth processes essential for development. Thus, several new postulated interactions, operating at the perception and early transduction stages of the signaling cascade leading to auxin-mediated bending, were proposed to be examined in snapdragon spikes and oat shoot pulvini, according to the following research goals: 1) Establish the role of amyloplasts as gravireceptors in shoots; 2) Investigate gravity-induced changes in the integrity of shoot actin cytoskeleton (CK); 3) Study the cellular interactions among actin CK, statoliths and cell membranes (endoplasmic reticulum - ER, plasma membrane - PM) during shoot graviperception; 4) Examine mediation of graviperception by modulations of cytosolic calcium - [Ca2+]cyt, and other second messengers (protein phosphorylation, inositol 1,4,5-trisphosphate - IP3). Revisions: 1) Model system: in addition to snapdragon (Antirrhinum majus L.) spikes and oat (Avena sativa) shoot pulvini, the model system of maize (Zea mays) primary roots was targeted to confirm a more general mechanism for graviperception. 2) Research topic: brassinolide, which were not included in the original plan, were examined for their regulatory role in gravity perception and signal transduction in roots, in relation to auxin and ethylene. Background to the topic: The negative gravitropic response of shoots is a complex multi-step process that requires the participation of various cellular components acting in succession or in parallel. Most of the long-lasting studies regarding the link between graviperception and cellular components were focused mainly on roots, and there are relatively few reports on shoot graviperception. Our previous project has successfully characterized several key events occurring during shoot bending of cut flowers and oat pulvini, including amyloplast displacement, hormonal interactions and differential growth analysis. Based on this evidence, the present project has focused on studying the initial graviperception process in flowering stems and cereal shoots. Major conclusions and achievements: 1) The actin and not the microtubule (MT) CK is involved in the graviperception of snapdragon shoots. 2) Gravisensing, exhibited by amyloplast displacement, and early transduction events (auxin redistribution) in the gravitropic response of snapdragon spikes are mediated by the acto-myosin complex. 3) MTs are involved in stem directional growth, which occurs during gravitropism of cut snapdragon spikes, but they are not necessary for the gravity-induced differential growth. 4) The role of amyloplasts as gravisensors in the shoot endodermis was demonstrated for both plant systems. 5) A gravity-induced increase in IP.

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Sengupta-Gopalan, Champa, Shmuel Galili, and Rachel Amir. Improving Methionine Content in Transgenic Forage Legumes. United States Department of Agriculture, February 2001. http://dx.doi.org/10.32747/2001.7580671.bard.

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Leguminous forage crops are high in proteins but deficient in S- amino acids. It has been shown that both wool quality and milk production can be limited by the post-ruminal supply of sulfur-containing amino acids. Efforts to use conventional plant breeding and cell selection techniques to increase the S-amino acid content of alfalfa have met with little success. With the objective to increase the S-amino acid content of forage legumes, the goal of this project was to co- express the methionine rich zein genes from corn along with a gene for a key enzyme in methionine biosynthesis, aspartate kinase(AK). The zeins are seed storage proteins from corn and are groupec into four distinct classes based on their amino acid sequence homologies. The b-zein (15kd) and the 6zein (10kD and 18kD) have proportionately high levels of methionine (10%, 22% and 28%, respectively). Initial studies from our lab had shown that while the 15kD zein accumulated to high levels in vegetative tissues of transgenic tobacco the l0kD zein did not. However, co-expression of the 10kD zein with the 15kD zein genes in tobacco showed stabilization of the 10kD zein and the co-localization of the 10kD and 15kD zein proteins in unique ER derived protein bodies. AK is the key enzyme for producing carbon skeletons for all amino acids of the aspartate family including methionine. It is, however, regulated by end-product feedback inhibition. The specific objectives of this proposal were: i. to co-express the 15kD zein with the 10/18kD zein genes in alfalfa in order to enhance the level of accumulation of the 10/18kD zein; ii. to increase methionine pools by expressing a feedback insensitive AK gene in transformants co-expressing the 15kD and 10/18kD zein genes. The Israeli partners were successful in expressing the AK gene in alfalfa which resulted in an increase in free and bound threonine but not in methionine (Galili et al., 2000). Since our target was to increase methionine pools, we changed our second objective to replace the AK gene with the gene for cystathionine gamma synthase (CGS) in the co-expression studies. The first methionine specific reaction is catalyzed by CGS. An additional objective was to develop a transformation system for Berseem clover, and to introduce the appropriate gene constructs into it with the goal of improving their methionine content. Genes for the 15kD zein along with the genes for either the 10kD or 18kD zein have been introduced into the same alfalfa plant both by sexual crosses and by re-transformation. Analysis of these zein co-expressors have shown that both the IOkD and 18kD zein levels go up 5 to 10 fold when co-expressed with the 15kD zein (Bagga et al., MS in preparation). Incubation of the leaves of transgenic alfalfa co-expressing the 15kD and 10kD zein genes, in the rumen of cows have shown that the zein proteins are stable in the rumen. To increase the level of zein accumulation in transgenic alfalfa different promoters have been used to drive the zein genes in alfalfa and we have concluded that the CaMV 35S promoter is superior to the other strong leaf -specific promoters. By feeding callus tissue of alfalfa plants co-expressing the 15kD and 10kD zein genes with methionine and its precursors, we have shown that the zein levels could be significantly enhanced by increasing the methionine pools. We have now introduced the CGS gene (from Arabidopsis; kindly provided to us by Dr. Leustek), into the 15kD zein transformants and experiments are in progress to check if the expression of the CGS gene indeed increases the level of zein accumulation in alfalfa. We were not successful in developing a transformation protocol for Berseem clover.

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