Journal ArticleProc Natl Acad Sci U S A · August 6, 2024
Invasive fungal diseases are a major threat to human health, resulting in more than 1.5 million annual deaths worldwide. The arsenal of antifungal therapeutics remains limited and is in dire need of drugs that target additional biosynthetic pathways that a ...
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Journal ArticleCurrent opinion in structural biology · August 2024
Cryogenic electron tomography (cryo-ET), a method that enables the viewing of biomolecules in near-native environments at high resolution, is rising in accessibility and applicability. Over the past several years, once slow sample preparation and data coll ...
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Journal ArticleBiological imaging · January 2024
Cryo-electron microscopy (cryo-EM) is an imaging technique that allows the visualization of proteins and macromolecular complexes at near-atomic resolution. The low electron doses used to prevent radiation damage to the biological samples result in images ...
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Journal ArticleNature methods · December 2023
Single-particle cryo-electron tomography is an emerging technique capable of determining the structure of proteins imaged within the native context of cells at molecular resolution. While high-throughput techniques for sample preparation and tilt-series ac ...
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Journal ArticleNature communications · August 2023
The human endogenous retrovirus K (HERV-K) is the most recently acquired endogenous retrovirus in the human genome and is activated and expressed in many cancers and amyotrophic lateral sclerosis. We present the immature HERV-K capsid structure at 3.2 Å re ...
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Journal ArticlebioRxiv · June 7, 2023
A significant part of the human genome consists of endogenous retroviruses sequences. Human endogenous retrovirus K (HERV-K) is the most recently acquired endogenous retrovirus, is activated and expressed in many cancers and amyotrophic lateral sclerosis a ...
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Journal ArticleNat Commun · May 15, 2023
Antibody affinity maturation enables adaptive immune responses to a wide range of pathogens. In some individuals broadly neutralizing antibodies develop to recognize rapidly mutating pathogens with extensive sequence diversity. Vaccine design for pathogens ...
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Journal ArticleNat Commun · March 20, 2023
SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that SPY is distinct from ER-localized POFUTs and contains N-terminal tetr ...
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Journal ArticleBiological imaging · January 2023
Single-particle cryo-electron microscopy (cryo-EM) is a powerful imaging modality capable of visualizing proteins and macromolecular complexes at near-atomic resolution. The low electron-doses used to prevent radiation damage to the biological samples, how ...
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Journal ArticleElife · August 23, 2022
Finding the conditions to stabilize a macromolecular target for imaging remains the most critical barrier to determining its structure by cryo-electron microscopy (cryo-EM). While automation has significantly increased the speed of data collection, specime ...
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Journal ArticleActa crystallographica. Section D, Structural biology · July 2022
Tomographic reconstruction of frozen-hydrated specimens followed by extraction and averaging of sub-tomograms has successfully been used to determine the structure of macromolecules in their native environment at resolutions that are high enough to reveal ...
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Journal ArticleComputer methods and programs in biomedicine · June 2022
Background and objectiveOne of the strengths of single-particle cryo-EM compared to other structural determination techniques is its ability to image heterogeneous samples containing multiple molecular species, different oligomeric states or disti ...
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Journal ArticleCell Rep · May 24, 2022
Protein ubiquitination is an essential process that rapidly regulates protein synthesis, function, and fate in dynamic environments. Within its non-proteolytic functions, we showed that K63-linked polyubiquitinated conjugates heavily accumulate in yeast ce ...
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Journal ArticleNature · May 2022
NPR1 is a master regulator of the defence transcriptome induced by the plant immune signal salicylic acid1-4. Despite the important role of NPR1 in plant immunity5-7, understanding of its regulatory mechanisms has been hindered by a lack of structural info ...
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ConferenceProceedings - 2022 IEEE/CVF Winter Conference on Applications of Computer Vision, WACV 2022 · January 1, 2022
Deep learning-based object detection methods have shown promising results in various fields ranging from autonomous driving to video surveillance where input images have relatively high signal-to-noise ratios (SNR). On low SNR images such as biological ele ...
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ConferenceLecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics) · January 1, 2022
Cryo-electron tomography (CET) combined with sub-volume averaging (SVA), is currently the only imaging technique capable of de-termining the structure of proteins imaged inside cells at molecular reso-lution. To obtain high-resolution reconstructions, sub- ...
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Journal ArticleProc Natl Acad Sci U S A · August 6, 2024
Invasive fungal diseases are a major threat to human health, resulting in more than 1.5 million annual deaths worldwide. The arsenal of antifungal therapeutics remains limited and is in dire need of drugs that target additional biosynthetic pathways that a ...
Full textLink to itemCite
Journal ArticleCurrent opinion in structural biology · August 2024
Cryogenic electron tomography (cryo-ET), a method that enables the viewing of biomolecules in near-native environments at high resolution, is rising in accessibility and applicability. Over the past several years, once slow sample preparation and data coll ...
Full textCite
Journal ArticleBiological imaging · January 2024
Cryo-electron microscopy (cryo-EM) is an imaging technique that allows the visualization of proteins and macromolecular complexes at near-atomic resolution. The low electron doses used to prevent radiation damage to the biological samples result in images ...
Full textCite
Journal ArticleNature methods · December 2023
Single-particle cryo-electron tomography is an emerging technique capable of determining the structure of proteins imaged within the native context of cells at molecular resolution. While high-throughput techniques for sample preparation and tilt-series ac ...
Full textCite
Journal ArticleNature communications · August 2023
The human endogenous retrovirus K (HERV-K) is the most recently acquired endogenous retrovirus in the human genome and is activated and expressed in many cancers and amyotrophic lateral sclerosis. We present the immature HERV-K capsid structure at 3.2 Å re ...
Full textCite
Journal ArticlebioRxiv · June 7, 2023
A significant part of the human genome consists of endogenous retroviruses sequences. Human endogenous retrovirus K (HERV-K) is the most recently acquired endogenous retrovirus, is activated and expressed in many cancers and amyotrophic lateral sclerosis a ...
Full textLink to itemCite
Journal ArticleNat Commun · May 15, 2023
Antibody affinity maturation enables adaptive immune responses to a wide range of pathogens. In some individuals broadly neutralizing antibodies develop to recognize rapidly mutating pathogens with extensive sequence diversity. Vaccine design for pathogens ...
Full textLink to itemCite
Journal ArticleNat Commun · March 20, 2023
SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that SPY is distinct from ER-localized POFUTs and contains N-terminal tetr ...
Full textOpen AccessLink to itemCite
Journal ArticleBiological imaging · January 2023
Single-particle cryo-electron microscopy (cryo-EM) is a powerful imaging modality capable of visualizing proteins and macromolecular complexes at near-atomic resolution. The low electron-doses used to prevent radiation damage to the biological samples, how ...
Full textCite
Journal ArticleElife · August 23, 2022
Finding the conditions to stabilize a macromolecular target for imaging remains the most critical barrier to determining its structure by cryo-electron microscopy (cryo-EM). While automation has significantly increased the speed of data collection, specime ...
Full textLink to itemCite
Journal ArticleActa crystallographica. Section D, Structural biology · July 2022
Tomographic reconstruction of frozen-hydrated specimens followed by extraction and averaging of sub-tomograms has successfully been used to determine the structure of macromolecules in their native environment at resolutions that are high enough to reveal ...
Full textCite
Journal ArticleComputer methods and programs in biomedicine · June 2022
Background and objectiveOne of the strengths of single-particle cryo-EM compared to other structural determination techniques is its ability to image heterogeneous samples containing multiple molecular species, different oligomeric states or disti ...
Full textCite
Journal ArticleCell Rep · May 24, 2022
Protein ubiquitination is an essential process that rapidly regulates protein synthesis, function, and fate in dynamic environments. Within its non-proteolytic functions, we showed that K63-linked polyubiquitinated conjugates heavily accumulate in yeast ce ...
Full textLink to itemCite
Journal ArticleNature · May 2022
NPR1 is a master regulator of the defence transcriptome induced by the plant immune signal salicylic acid1-4. Despite the important role of NPR1 in plant immunity5-7, understanding of its regulatory mechanisms has been hindered by a lack of structural info ...
Full textLink to itemCite
ConferenceProceedings - 2022 IEEE/CVF Winter Conference on Applications of Computer Vision, WACV 2022 · January 1, 2022
Deep learning-based object detection methods have shown promising results in various fields ranging from autonomous driving to video surveillance where input images have relatively high signal-to-noise ratios (SNR). On low SNR images such as biological ele ...
Full textCite
ConferenceLecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics) · January 1, 2022
Cryo-electron tomography (CET) combined with sub-volume averaging (SVA), is currently the only imaging technique capable of de-termining the structure of proteins imaged inside cells at molecular reso-lution. To obtain high-resolution reconstructions, sub- ...
Full textCite
Journal ArticleCell · May 27, 2021
Natural antibodies (Abs) can target host glycans on the surface of pathogens. We studied the evolution of glycan-reactive B cells of rhesus macaques and humans using glycosylated HIV-1 envelope (Env) as a model antigen. 2G12 is a broadly neutralizing Ab (b ...
Full textOpen AccessLink to itemCite
Journal ArticleNat Commun · March 30, 2021
Featured Publication
Tomographic reconstruction of cryopreserved specimens imaged in an electron microscope followed by extraction and averaging of sub-volumes has been successfully used to derive atomic models of macromolecules in their biological environment. Eliminating bio ...
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Journal ArticleMol Cell · January 7, 2021
The bacterium Francisella tularensis (Ft) is one of the most infectious agents known. Ft virulence is controlled by a unique combination of transcription regulators: the MglA-SspA heterodimer, PigR, and the stress signal, ppGpp. MglA-SspA assembles with th ...
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ConferenceProceedings - Applied Imagery Pattern Recognition Workshop · January 1, 2021
Specimen optimization is currently one of the main limiting steps in the cryo-electron microscopy (EM) structure determination pipeline. The ideal specimen is a molecule-thin layer of macromolecules in solution frozen on top of a holey membrane stabilized ...
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ConferenceProceedings - Applied Imagery Pattern Recognition Workshop · January 1, 2021
Particle picking is a critical step in the single-particle cryo-electron microscopy (EM) structure determination pipeline. In order to successfully obtain the 3D reconstruction of a macro-molecular complex, hundreds of thousands of particles need to be acc ...
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Journal Article · November 22, 2020
Single-particle cryo-electron microscopy (cryo-EM) is an emerging imaging
modality capable of visualizing proteins and macro-molecular complexes at
near-atomic resolution. The low electron-doses used to prevent sample radiation
damage, result in images whe ...
Link to itemCite
Journal ArticleProc Natl Acad Sci U S A · September 8, 2020
Subpopulations of ribosomes are responsible for fine tuning the control of protein synthesis in dynamic environments. K63 ubiquitination of ribosomes has emerged as a new posttranslational modification that regulates protein synthesis during cellular respo ...
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Journal ArticleIEEE signal processing magazine · March 2020
In recent years, an abundance of new molecular structures have been elucidated using cryo-electron microscopy (cryo-EM), largely due to advances in hardware technology and data processing techniques. Owing to these new exciting developments, cryo-EM was se ...
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Journal ArticleNat Commun · January 24, 2020
The trimeric HIV-1 Envelope protein (Env) mediates viral-host cell fusion via a network of conformational transitions, with allosteric elements in each protomer orchestrating host receptor-induced exposure of the co-receptor binding site and fusion element ...
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Journal ArticleInverse Problems · January 1, 2020
Single-particle cryo-electron microscopy (EM) has become a popular technique for determining the structure of challenging biomolecules that are inaccessible to other technologies. Recent advances in automation, both in data collection and data processing, ...
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Journal ArticleICASSP, IEEE International Conference on Acoustics, Speech and Signal Processing - Proceedings · September 10, 2018
Multi-image alignment, bringing a group of images into common register, is an ubiquitous problem and the first step of many applications in a wide variety of domains. As a result, a great amount of effort is being invested in developing efficient multi-ima ...
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Journal ArticleNature · September 2018
In the PDF version of this Article, owing to a typesetting error, an incorrect figure was used for Extended Data Fig. 5; the correct figure was used in the HTML version. This has been corrected online. ...
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Journal ArticleeLife · August 2018
Voltage-activated potassium (Kv) channels open to conduct K+ ions in response to membrane depolarization, and subsequently enter non-conducting states through distinct mechanisms of inactivation. X-ray structures of detergent-solubilized Kv chan ...
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Journal ArticleStructure (London, England : 1993) · June 2018
Featured Publication
The advent of direct electron detectors has enabled the routine use of single-particle cryo-electron microscopy (EM) approaches to determine structures of a variety of protein complexes at near-atomic resolution. Here, we report the development of methods ...
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Journal ArticleNature · June 2018
Featured Publication
G-protein-coupled receptors comprise the largest family of mammalian transmembrane receptors. They mediate numerous cellular pathways by coupling with downstream signalling transducers, including the hetrotrimeric G proteins Gs (stimulatory) and ...
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Journal ArticleCell · October 2017
Featured Publication
Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. A central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or ...
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Journal ArticleIEEE Transactions on Signal Processing · November 1, 2016
Featured Publication
The performance of multiimage alignment, bringing different images into one coordinate system, is critical in many applications with varied signal-to-noise ratio (SNR) conditions. A great amount of effort is being invested into developing methods to solve ...
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Journal ArticleMol Pharmacol · July 2016
The multidrug transporter P-glycoprotein (P-gp, ABCB1) is an ATP-dependent pump that mediates the efflux of structurally diverse drugs and xenobiotics across cell membranes, affecting drug pharmacokinetics and contributing to the development of multidrug r ...
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Journal ArticleMol Pharmacol · June 2016
Cryo-electron microscopy (cryo-EM) methods are now being used to determine structures at near-atomic resolution and have great promise in molecular pharmacology, especially in the context of mapping the binding of small-molecule ligands to protein complexe ...
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Journal ArticleCell · June 2016
Featured Publication
Recent advances in single-particle cryoelecton microscopy (cryo-EM) are enabling generation of numerous near-atomic resolution structures for well-ordered protein complexes with sizes ≥ ∼200 kDa. Whether cryo-EM methods are equally useful for high-resoluti ...
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Journal ArticlemBio · March 2016
UnlabelledInfluenza viruses expressing chimeric hemagglutinins (HAs) are important tools in the quest for a universal vaccine. Using cryo-electron tomography, we have determined the structures of a chimeric HA variant that comprises an H1 stalk an ...
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Journal ArticleCell · February 11, 2016
CorA, the major Mg(2+) uptake system in prokaryotes, is gated by intracellular Mg(2+) (KD ∼ 1-2 mM). X-ray crystallographic studies of CorA show similar conformations under Mg(2+)-bound and Mg(2+)-free conditions, but EPR spectroscopic studies reveal large ...
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Journal ArticleScience (New York, N.Y.) · February 2016
p97 is a hexameric AAA+ adenosine triphosphatase (ATPase) that is an attractive target for cancer drug development. We report cryo-electron microscopy (cryo-EM) structures for adenosine diphosphate (ADP)-bound, full-length, hexameric wild-type p97 in the p ...
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Journal ArticleScience (New York, N.Y.) · June 2015
Cryo-electron microscopy (cryo-EM) is rapidly emerging as a powerful tool for protein structure determination at high resolution. Here we report the structure of a complex between Escherichia coli β-galactosidase and the cell-permeant inhibitor phenylethyl ...
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Journal ArticleNature · October 2014
Ionotropic glutamate receptors are ligand-gated ion channels that mediate excitatory synaptic transmission in the vertebrate brain. To gain a better understanding of how structural changes gate ion flux across the membrane, we trapped rat AMPA (α-amino-3-h ...
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Journal ArticleJournal of virology · September 2014
The Ebola virus glycoprotein mucin-like domain (MLD) is implicated in Ebola virus cell entry and immune evasion. Using cryo-electron tomography of Ebola virus-like particles, we determined a three-dimensional structure for the full-length glycoprotein in a ...
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Journal ArticleProceedings of the National Academy of Sciences of the United States of America · August 2014
We report the solution structure of Escherichia coli β-galactosidase (∼465 kDa), solved at ∼3.2-Å resolution by using single-particle cryo-electron microscopy (cryo-EM). Densities for most side chains, including those of residues in the active site, and a ...
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Journal ArticleNat Struct Mol Biol · December 2013
The activation of trimeric HIV-1 envelope glycoprotein (Env) by its binding to the cell-surface receptor CD4 and co-receptors (CCR5 or CXCR4) represents the first of a series of events that lead to fusion between viral and target-cell membranes. Here, we p ...
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Journal ArticleJournal of virology · June 2013
We describe cryo-electron microscopic studies of the interaction between the ectodomain of the trimeric HIV-1 envelope glycoprotein (Env) and Z13e1, a broadly neutralizing antibody that targets the membrane-proximal external region (MPER) of the gp41 subun ...
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Journal ArticleProc Natl Acad Sci U S A · April 9, 2013
Glutamate receptor ion channels are membrane proteins that mediate excitatory synaptic transmission in the central nervous system of vertebrates. Insight into molecular mechanisms underlying glutamate receptor gating is limited by lack of structural inform ...
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Journal ArticleJ Struct Biol · February 2013
The limitation of using low electron doses in non-destructive cryo-electron tomography of biological specimens can be partially offset via averaging of aligned and structurally homogeneous subsets present in tomograms. This type of sub-volume averaging is ...
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Journal ArticleFEBS J · January 2013
Cryo-electron microscopy (cryo-EM) is increasingly becoming a mainstream technology for studying the architecture of cells, viruses and protein assemblies at molecular resolution. Recent developments in microscope design and imaging hardware, paired with e ...
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Journal ArticleStructure (London, England : 1993) · December 2012
Cryo-electron microscopy (cryo-EM) is a powerful technique for 3D structure determination of protein complexes by averaging information from individual molecular images. The resolutions that can be achieved with single-particle cryo-EM are frequently limit ...
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Journal ArticleJ Struct Biol · May 2012
We have previously used cryo-electron tomography combined with sub-volume averaging and classification to obtain 3D structures of macromolecular assemblies in cases where a single dominant species was present, and applied these methods to the analysis of a ...
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Journal ArticlePLoS Pathog · 2012
HIV-1 infection begins with the binding of trimeric viral envelope glycoproteins (Env) to CD4 and a co-receptor on target T-cells. Understanding how these ligands influence the structure of Env is of fundamental interest for HIV vaccine development. Using ...
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Journal ArticleJ Vis Exp · December 1, 2011
Since its discovery nearly 30 years ago, more than 60 million people have been infected with the human immunodeficiency virus (HIV) (www.usaid.gov). The virus infects and destroys CD4+ T-cells thereby crippling the immune system, and causing an acquired im ...
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Journal ArticleJ Virol · December 2011
The trimeric envelope glycoprotein (Env) spikes displayed on the surfaces of simian immunodeficiency virus (SIV) and human immunodeficiency virus type 1 (HIV-1) virions are composed of three heterodimers of the viral glycoproteins gp120 and gp41. Although ...
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Journal ArticleProc Natl Acad Sci U S A · July 12, 2011
The initial step in HIV-1 infection occurs with the binding of cell surface CD4 to trimeric HIV-1 envelope glycoproteins (Env), a heterodimer of a transmembrane glycoprotein (gp41) and a surface glycoprotein (gp120). The design of soluble versions of trime ...
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Journal ArticlePLoS Pathog · December 23, 2010
The initial step in target cell infection by human, and the closely related simian immunodeficiency viruses (HIV and SIV, respectively) occurs with the binding of trimeric envelope glycoproteins (Env), composed of heterodimers of the viral transmembrane gl ...
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Journal ArticleCurrent opinion in structural biology · August 2009
The vast majority of membrane protein complexes of biological interest cannot be purified to homogeneity, or removed from a physiologically relevant context without loss of function. It is therefore not possible to easily determine the 3D structures of the ...
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Journal ArticleNature · September 4, 2008
The envelope glycoproteins (Env) of human and simian immunodeficiency viruses (HIV and SIV, respectively) mediate virus binding to the cell surface receptor CD4 on target cells to initiate infection. Env is a heterodimer of a transmembrane glycoprotein (gp ...
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Journal ArticleJournal of structural biology · June 2008
Strategies for the determination of 3D structures of biological macromolecules using electron crystallography and single-particle electron microscopy utilize powerful tools for the averaging of information obtained from 2D projection images of structurally ...
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Journal Article2007 4th IEEE International Symposium on Biomedical Imaging: From Nano to Macro - Proceedings · November 27, 2007
In limited angle tomography, with applications such as electron microscopy, medical imaging, and industrial testing, the object of interest is scanned over a limited angular range, which is less than the full 180° mathematically required for density recons ...
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Journal Article2007 4th IEEE International Symposium on Biomedical Imaging: From Nano to Macro - Proceedings · November 27, 2007
Electron tomography provides opportunities to determine diree-dimensional cellular architecture at resolutions high enough to identify individual macromolecules such as proteins. Image analysis of such data poses a challenging problem due to the extremely ...
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Journal ArticleCurrent opinion in structural biology · October 2007
Understanding the molecular architectures of enveloped and complex viruses is a challenging frontier in structural biology. In these viruses, the structural and compositional variation from one viral particle to another generally precludes the use of eithe ...
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Journal ArticlePLoS pathogens · May 2007
The envelope glycoproteins of primate lentiviruses, including human and simian immunodeficiency viruses (HIV and SIV), are heterodimers of a transmembrane glycoprotein (usually gp41), and a surface glycoprotein (gp120), which binds CD4 on target cells to i ...
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Conference2006 3rd IEEE International Symposium on Biomedical Imaging: From Nano to Macro - Proceedings · November 17, 2006
A general framework for the automatic segmentation of anatomical structures from Diffusion Tensor MRI is presented here. We adopt an energy based approach to segmentation assuming a piecewise-smooth image model that allows tensors to change orientation ins ...
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Journal ArticleJournal of Computational Physics · March 1, 2006
In this note we present an implementation of the fast marching algorithm for solving Eikonal equations that in practice reduces the original run-time from O(N log N) to linear. This lower run-time cost is obtained while keeping an error bound of the same o ...
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Journal ArticleIEEE Geoscience and Remote Sensing Letters · January 1, 2006
A framework for geometric regularization of elevation maps is introduced in this letter. The framework takes into account errors in the data, which form part of standard elevation maps specifications, as well as possible additional user/application-depende ...
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Journal ArticleProceedings - International Conference on Image Processing, ICIP · December 1, 2005
We present an algorithm for tracking moving objects using intrinsic minimal surfaces which handles particularly well the presence of severe and total occlusions even in the presence of weak object boundaries. We adopt an edge based approach and find the se ...
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Journal ArticleIEEE transactions on image processing : a publication of the IEEE Signal Processing Society · September 2005
Electron tomography allows for the determination of the three-dimensional structures of cells and tissues at resolutions significantly higher than that which is possible with optical microscopy. Electron tomograms contain, in principle, vast amounts of inf ...
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Journal ArticleGraphical Models · July 1, 2005
A paradigm for automatic three-dimensional shape and geometry rendering from multiple images is introduced in this paper. In particular, non-photorealistic rendering (NPR) techniques in the style of pen-and-ink illustrations are addressed, while the underl ...
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Journal ArticleProceedings - International Conference on Image Processing, ICIP · December 1, 2004
A new paradigm for automatic non-photorealistic rendering (NPR) is introduced in this paper. Existing NPR approaches can be categorized in two groups depending on the type of input they use: image based and object based. Using multiple images as input to t ...
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Journal Article2004 2nd IEEE International Symposium on Biomedical Imaging: Macro to Nano · December 1, 2004
Electron tomography allows determination of the three-dimensional structures of cells and tissues at resolutions significantly higher than is possible with optical microscopy. Electron tomograms contain, in principle, vast amounts of information on the loc ...
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Journal ArticleHuman brain mapping · September 2001
In this study, a computational optimal system for the generation of curves on triangulated surfaces representing 3D brains is described. The algorithm is based on optimally computing geodesics on the triangulated surfaces following Kimmel and Sethian ([199 ...
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ConferenceLecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics) · January 1, 2001
We study and evaluate an automatic fingerprint classification algorithm that we apply over the fully manual identification system being used by the Dirección Nacional de Identificación Civil (DNIC). To be compatible with the existing system and provide a g ...
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