• His laboratory made several contributions in these fields of research: Differentiation and genetic manipulation of human embryonic stem cells: In 2000, his laboratory pioneered the first demonstration of both spontaneous differentiation of human embryonic stem cells into embryoid bodies and direct differentiation into more than ten cell types. (wikipedia.org)
  • In addition, his laboratory was the first to demonstrate genetic manipulation of human embryonic stem cells. (wikipedia.org)
  • Haploid human embryonic stem cells and genome-wide screenings: Benvenisty's lab was the first to generate haploid human embryonic stem cells (having half of the chromosomes). (wikipedia.org)
  • Wutz, A. Haploid mouse embryonic stem cells: rapid genetic screening and germline transmission. (nature.com)
  • Leeb, M. & Wutz, A. Derivation of haploid embryonic stem cells from mouse embryos. (nature.com)
  • Forward and reverse genetics through derivation of haploid mouse embryonic stem cells. (nature.com)
  • Generation of genetically modified mice by oocyte injection of androgenetic haploid embryonic stem cells. (nature.com)
  • Androgenetic haploid embryonic stem cells produce live transgenic mice. (nature.com)
  • Genetic modification and screening in rat using haploid embryonic stem cells. (nature.com)
  • Generation of haploid embryonic stem cells from Macaca fascicularis monkey parthenotes. (nature.com)
  • Derivation and differentiation of haploid human embryonic stem cells. (nature.com)
  • Depending on the source, stem cells can be classified into two broad categories i.e. embryonic stem cells that are derived from embryos and non-embryonic stem cells that are derived from adult and fetal tissues. (benthamscience.com)
  • The non-embryonic stem cells like adult stem cells are in clinical use for many years and embryonic stem cells are now emerging as an alternative source for the same purpose with huge potentials in drug discovery and toxicological studies. (benthamscience.com)
  • Embryonic stem cells (ESCs) can grow infinitely and give rise to all types of cells in human body, thus of tremendous therapeutic potentials for a variety of diseases, such as Parkinson's disease, spinal cord injury, and diabetes. (benthamscience.com)
  • Mouse nuclear transfer embryonic stem cells (NT-ESCs) were first established in 2000, and then proved to be able to differentiate either in vivo or in vitro, and give rise to individual tissues through germ line transmission or tetraploid complementation. (benthamscience.com)
  • A unique and essential property of embryonic stem cells is the ability to self-renew and differentiate into multiple cell lineages. (biomedcentral.com)
  • However, the possible differences in proliferation and differentiation capabilities among independently-derived human embryonic stem cells (hESCs) are not well known because of insufficient characterization. (biomedcentral.com)
  • All cell lines differentiated into three embryonic germ lineages in embryoid bodies and into neural cell lineages when cultured in neural differentiation medium. (biomedcentral.com)
  • Human embryonic stem cells (hESCs) possess the ability to self-renew in an undifferentiated state in culture while retaining the ability to differentiate into all of the cell types in the human body. (biomedcentral.com)
  • In the present study, a side-by-side comparison of the ability to maintain an undifferentiated state and to self-renew under standard conditions, the ability to spontaneously differentiate into cell types of three germ layers in embryonic bodies, and directed differentiation under neural differentiation-promoting conditions was made between three NIH registered hESC lines I3, I6 and BG01V. (biomedcentral.com)
  • We discuss these properties with examples both from the hematopoietic and embryonic stem cell (ESC) systems. (lu.se)
  • Mouse embryonic fibroblast cells (MEFs) carrying an Oct4-GFP reporter were derived from day 13.5 embryos of the OG2 transgenic mouse strain and were cultured in Fibro Gro Low Serum (F.Gro) medium (Millipore). (molcells.org)
  • In vitro stem cell models that replicate human gastrulation have been generated, but they lack the essential extraembryonic cells needed for embryonic development, morphogenesis, and patterning. (bvsalud.org)
  • Here, we describe a robust and efficient method that prompts human extended pluripotent stem cells to self-organize into embryo-like structures, termed peri-gastruloids, which encompass both embryonic (epiblast) and extraembryonic (hypoblast) tissues. (bvsalud.org)
  • Although peri-gastruloids are not viable due to the exclusion of trophoblasts, they recapitulate critical stages of human peri-gastrulation development, such as forming amniotic and yolk sac cavities, developing bilaminar and trilaminar embryonic discs, specifying primordial germ cells, initiating gastrulation, and undergoing early neurulation and organogenesis. (bvsalud.org)
  • Embryonic stem cell technology is still at a preliminary research stage and announcements about its potential may be premature. (edu.au)
  • Experts from around the world are assessing the difficult issue of the extent to which embryonic stem cell research should be allowed to proceed, and to date there is little international consensus on this matter. (edu.au)
  • How, then, should embryonic stem cell research be regulated in Australia? (edu.au)
  • In this article we examine embryonic stem cell research and explore the current regulatory framework associated with this research in Australia, with particular reference to the Andrews Report . (edu.au)
  • Launch Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), possess variations within their capability to differentiate1. (tam-receptor.com)
  • Stem cells are not specialized and the process of their specialization is called differentiation. (benthamscience.com)
  • 1 Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan. (jci.org)
  • and 3) the responses to directed neural differentiation was made between three NIH registered hES cell lines I3 (TE03), I6 (TE06) and BG01V. (biomedcentral.com)
  • Undifferentiated I3 cells grew significantly slower but their differentiation potential was greater than I6 and BG01V. (biomedcentral.com)
  • Under the same neural differentiation-promoting conditions, the ability of each cell line to differentiate into neural progenitors varied. (biomedcentral.com)
  • Stem and progenitor cell populations are often heterogeneous, which may reflect stem cell subsets that express subtly different properties, including different propensities for lineage selection upon differentiation, yet remain able to interconvert. (lu.se)
  • A key challenge is to understand how state, but must also afford flexibility in cell-fate choice to permit the different cell-fate options confronting stem and progenitor cell-type diversification and differentiation in response to cells are selected and coordinated such that adoption of a given intrinsic cues or extrinsic signals. (lu.se)
  • Evidence the fate of stem cells has broad ramifications for biomedical suggests that during development or differentiation, cells make science from elucidating the causes of cancer to the use of very precise transitions between apparently stable ``network stem cells in regenerative medicine. (lu.se)
  • The AF form of SALSA had a more intact structure and contained peptides from the zona pellucida domain, which is involved in cell differentiation and oligomerization. (researchgate.net)
  • Sagi, I. & Benvenisty, N. Haploidy in humans: an evolutionary and developmental perspective. (nature.com)
  • To take human organ generation via BC and transplantation to the next step, we reviewed current emerging organ generation technologies and the associated efficiency of chimera formation in human cells from the standpoint of developmental biology. (frontiersin.org)
  • These developmental defects have been attributed to incomplete reprogramming of the somatic nuclei by the cloning process. (biomedcentral.com)
  • Developmental defects, including abnormalities in cloned fetuses and placentas, in addition to high rates of pregnancy loss and neonatal death have been encountered by every research team studying somatic cloning. (biomedcentral.com)
  • A key obstacle to progress is the systematic functional characterization of non-coding variants at scale, especially since enhancer activity is highly specific to cell type and developmental stage. (bvsalud.org)
  • Thus, genetic variants at enhancers are thought to contribute to developmental diseases by altering cell fate commitment. (bvsalud.org)
  • Under such circumstances, the idea of "therapeutic cloning" was proposed, indicating the generation of ESCs from SCNT embryos for therapeutic purpose. (benthamscience.com)
  • What is more, by deriving NT-ESCs from patient cells, the problem of immune rejection may be avoided. (benthamscience.com)
  • However, the derivation of human NT-ESCs goes with the destruction of clone embryos, leading to fierce ethical disputes. (benthamscience.com)
  • He is a founding member of the Israel Stem Cell Society, and he is the Founder and CSO of NewStem Ltd. Benvenisty's research projects focus on pluripotent stem cell biology, human genetic disorders, tissue engineering, genetic and epigenetic aberrations and cancer research. (wikipedia.org)
  • Understanding cell-fate decisions in stem cell populations is a major goal of modern biology. (lu.se)
  • Mechanochemical Principles of Spatial and Temporal Patterns in Cells and Tissues. (mpi-cbg.de)
  • Patterns are ubiquitous in living systems and underlie the dynamic organization of cells, tissues, and embryos. (mpi-cbg.de)
  • Some of these concerns, such as ensuring the welfare of research animals and obtaining appropriate consent for the use of human tissues, also apply to many other areas of research, but may require special consideration for research with human neural organoids, cell transplants, and chimeras. (nationalacademies.org)
  • Given the complexity of the human brain and the particularly human nature of many key symptoms of these disorders, especially psychiatric disorders, animal and cell culture models of the types currently used to investigate diseases of other organs and tissues are valuable but inadequate. (nationalacademies.org)
  • Stem cells may be derived from adult tissues but the most potent are extracted from developing human embryos. (edu.au)
  • This mode of cell death serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth. (lookformedical.com)
  • The Rappaport Prize for excellence in the field of biomedical research (2023) The Katzir Prize (2020) for exceptional achievements in life sciences, given triennial by FISEB/ILANIT. (wikipedia.org)
  • Cloning by nuclear transfer using mammalian somatic cells has enormous potential application. (biomedcentral.com)
  • With roughly 30,000 genes in mammalian genomes, fection with a vector encoding MyoD (Tapscott et al. (lu.se)
  • Induced pluripotent stem cells (iPSCs) are capable of unlimited self-renewal and can give rise to all three germ layers, thereby providing a new platform with which to study mammalian development and epigenetic reprogramming. (molcells.org)
  • Genetic and epigenetic stability of human pluripotent stem cells: The genetic and epigenetic instability of human pluripotent stem cells is a major characteristic of the cells affecting their tumorigenicity and their use in disease modeling. (wikipedia.org)
  • Benvenisty's lab was the first to study the chromosomal stability of human induce pluripotent stem cells, and developed methodologies to analyze genetic and epigenetic aberrations. (wikipedia.org)
  • However, iPSC generation may result in subtle epigenetic variations, such as the aberrant methylation of the Dlk1-Dio3 locus, among the clones, and this heterogeneity constitutes a major drawback to harnessing the full potential of iPSCs. (molcells.org)
  • This variability is certainly caused by hereditary and TGX-221 inhibition epigenetic distinctions that occur during derivation, induction, and following maintenance of PSCs2,3. (tam-receptor.com)
  • Kuldip S. Sidhu , " Frontiers in Pluripotent Stem Cells Research and Therapeutic Potentials Bench-to-Bedside ", Bentham Science Publishers (2012). (benthamscience.com)
  • The comparison of the unique properties and behavior of each individually derived cell line is critical in identifying the safe and efficacious lines for research and therapeutic use [ 3 , 13 ]. (biomedcentral.com)
  • As explained in Chapter 2 , human neural organoids, transplants, and chimeras provide new models for such conditions and may lead to new knowledge about brain development and function, the discovery of disease mechanisms, new therapeutic targets, and better screening of potential new treatments. (nationalacademies.org)
  • In Enhancing Evolution, leading bioethicist John Harris dismantles objections to genetic engineering, stem-cell research, designer babies, and cloning and makes an ethical case for biotechnology that is both forthright and rigorous. (philpapers.org)
  • Nissim Benvenisty is Professor of Genetics, the Herbert Cohn Chair in Cancer Research and the Director of "The Azrieli Center for Stem Cells and Genetic Research" at the Alexander Silberman Institute of Life Sciences, Hebrew University. (wikipedia.org)
  • His laboratory utilized these cells for genome-wide genetic screenings to analyze human development and disease. (wikipedia.org)
  • Awarded the "Azrieli Center for Stem Cells and Genetic Research" by the Azrieli Foundation (Canada) (2014). (wikipedia.org)
  • These differences may be associated with inherited variation in the sex, stage, quality and genetic background of embryos used for hESC line derivation, and/or changes acquired during passaging in culture. (biomedcentral.com)
  • It is also essential to understand how the inherited variation in the sex, stage, quality and genetic background of embryos, as well as environmental influences such as derivation methods and passage procedures can affect the ability of hES cell lines to self-renew and differentiate. (biomedcentral.com)
  • Directly comparing hES cell lines is challenging since all the genetic, environmental and methodological variables complicate the assessments. (biomedcentral.com)
  • Human enhancement, Harris argues, is a good thing--good morally, good for individuals, good as social policy, and good for a genetic heritage that needs serious improvement. (philpapers.org)
  • We perform a single-cell CRISPRi screen to assess the endogenous roles of 25 enhancers and putative cardiac target genes implicated in genetic studies of congenital heart defects (CHDs). (bvsalud.org)
  • explosion further, consider that a fictitious small genome with 2002) More recently and more dramatically, the potential for 260 genes would host the same number of combinations as cell state conversions is exemplified by the reprogramming of the number of atoms in the visible universe! (lu.se)
  • As a result, reprogramming somatic cells into iPSCs is normally along with a metabolic change from oxidative phosphorylation to glycolysis, concomitant with Mouse monoclonal to GST adjustments in function and framework of mitochondria16,17. (tam-receptor.com)
  • However, iPSC generation is a slow, inefficient process, and not all of the resulting iPSC clones are functionally equivalent. (molcells.org)
  • A classification of B-lymphocytes based on structurally or functionally different populations of cells. (lookformedical.com)
  • However, somatic cloning has been inefficient in all species in which live clones have been produced. (biomedcentral.com)
  • Malignant lymphoma composed of large B lymphoid cells whose nuclear size can exceed normal macrophage nuclei, or more than twice the size of a normal lymphocyte. (lookformedical.com)
  • He is a member of the steering committee of the International Stem Cell Initiative (ISCI), the Programme Board of the UK-RMP, and serves as the academic advisor for the International Society for Stem Cell Research (ISSCR). (wikipedia.org)
  • Automatic detection and segmentation of biological objects in 2D and 3D image data is central for countless biomedical research questions to be answered. (mpi-cbg.de)
  • These unique capabilities make hESCs a renewable source of a wide range of cell types for potential use in research and cell-based drug screening and therapies for many diseases. (biomedcentral.com)
  • These cells have been in high demand for use in basic and applied biomedical research. (biomedcentral.com)
  • Research involving human neural organoids, transplants, and chimeras has an ultimate goal of preventing and treating the great suffering caused by serious neurological and psychiatric conditions for which no effective treatment is available. (nationalacademies.org)
  • One such concern is the possibility of altering the capacities or consciousness of a research animal in ways that may blur the lines between human beings and nonhuman animals. (nationalacademies.org)
  • A main justification for carrying out research, both basic and translational, with human neural organoids, transplants, and chimeras is that it will help in the discovery of new ways to understand and treat neurological and psychiatric disorders, which, as discussed previously, cause immense suffering and for which treatments are ineffective or lacking. (nationalacademies.org)
  • Assisted reproductive technology (ART) and embryo research have posed many challenges to the different timeframes of science, ethics and law. (edu.au)
  • This issue was considered by the House of Representatives Standing Committee on Legal and Constitutional Affairs in its report entitled Human Cloning: Scientific, Ethical and Regulatory Aspects of Human Cloning and Stem Cell Research (hereafter the Andrews Report , after the Chair of the Committee, Mr Kevin Andrews, MP) released in September 2001. (edu.au)
  • Recent advances in human blastoids have opened new avenues for modeling early human development and implantation. (bvsalud.org)
  • However, a profound variation in colony morphology, growth rate, BrdU incorporation, and relative abundance of gene expression in undifferentiated and differentiated states of the cell lines was observed. (biomedcentral.com)
  • Similarly, GATA-1 has been shown to induce lineage switching expression values even if, for simplicity, we assume only ``on'' of committed cells in hematopoiesis, first in cell lines (Kulessa and ``off'' states for each gene. (lu.se)
  • In reality, gene somatic cells to a pluripotent cell state by a handful of transcrip- expression is graded, making the potential gene expression tion factors (Takahashi and Yamanaka, 2006). (lu.se)
  • Traditionally, selection of live PSCs with high pluripotency utilizes imaging methods that require fluorescent labeling of cells by immunostaining or gene transfection6,7. (tam-receptor.com)
  • The majority of mantle-cell lymphomas are associated with a t(11;14) translocation resulting in overexpression of the CYCLIN D1 gene (GENES, BCL-1). (lookformedical.com)
  • Ontology changes were based on recently published insights into the cellular and gross anatomy of these structures, and on new analyses of epithelial cell types present in the pelvic urethra and regions of the bladder. (atlas-d2k.org)
  • Fig. 1: Establishment and characterization of a genome-wide CRISPR-Cas9 screen in haploid hPSCs. (nature.com)
  • Because cattle are a species widely used for nuclear transfer studies, and more laboratories have succeeded in cloning cattle than any other specie, this review will be focused on somatic cell cloning of cattle. (biomedcentral.com)
  • Somatic cell cloning (cloning or nuclear transfer) is a technique in which the nucleus (DNA) of a somatic cell is transferred into an enucleated metaphase-II oocyte for the generation of a new individual, genetically identical to the somatic cell donor (Figure 1 ). (biomedcentral.com)
  • Currently, the efficiency for nuclear transfer is between 0-10%, i.e., 0-10 live births after transfer of 100 cloned embryos. (biomedcentral.com)
  • Various strategies have been employed to modify donor cells and the nuclear transfer procedure in attempts to improve the efficiency of nuclear transfer. (biomedcentral.com)
  • Although the efficiency of nuclear transfer has been dramatically improved from the initial success rate of one live clone born from 277 embryo transfers [ 1 ], none of the aforementioned efforts abolished the common problems associated with nuclear transfer. (biomedcentral.com)
  • A matured oocyte (c) is then enucleated (d) and a donor cell is transferred into the enucleated oocyte (e). (biomedcentral.com)
  • The report arose out of a recommendation for the Committee to review the report of the Australian Health Ethics Committee (AHEC) of the NHMRC entitled Scientific, Ethical and Regulatory Considerations Relevant to Cloning of Human Beings (hereafter the AHEC Report ). (edu.au)
  • Ontology changes include new structures, tissue layers and cell types within the LUT, external genitalia and lower reproductive structures. (atlas-d2k.org)
  • and the general public debate about reproductive cloning. (edu.au)
  • However, though BC is emerging as a potential organ transplant option, challenges regarding organ size scalability, immune system incompatibilities, long-term maintenance, potential evolutionary distance, or unveiled mechanisms between donor and host cells remain. (frontiersin.org)
  • At the organismal level, evolutionary distance is defined by genome-wide sequence homology between species. (frontiersin.org)
  • For instance, PSCs with low pluripotency may generate a people of somatic cells that might be polluted with undifferentiated or partially differentiated cells, which present a risk of tumor formation or low effectiveness after transplantation4,5. (tam-receptor.com)
  • Using our recently established haploid human pluripotent stem cells (hPSCs), we generated a genome-wide loss-of-function library targeting 18,166 protein-coding genes to define the essential genes in hPSCs. (nature.com)
  • hPSC-enriched essential genes mainly encode transcription factors and proteins related to cell-cycle and DNA-repair, revealing that a quarter of the nuclear factors are essential for normal growth. (nature.com)
  • Overall, we have constructed an atlas of essential and growth-restricting genes in hPSCs, revealing key aspects of cellular essentiality and providing a reference for future studies on human pluripotency. (nature.com)
  • Fig. 2: Analysis of cell-essential genes. (nature.com)
  • In terminally differentiated cell fate is coupled to appropriate regulation of the alternative cells, transcriptional networks must be stable and irreversible, pathways. (lu.se)
  • In this regard, emerging technologies of chimeric human organ production via blastocyst complementation (BC) holds great promise. (frontiersin.org)
  • The blastocyst can then be transferred to a recipient (h) and cloned animals are born after completion of gestation (i). (biomedcentral.com)
  • Additionally, we uncovered endometrial stromal effects in promoting trophoblast cell survival, proliferation, and syncytialization during co-culture with blastoids and blastocysts. (bvsalud.org)
  • Side-by-side single-cell RNA sequencing revealed similarities and differences in transcriptome profiles between pre-implantation blastoids and blastocysts, as well as post-implantation cultures, and uncovered a population resembling early migratory trophoblasts during co-culture with endometrial stromal cells. (bvsalud.org)
  • In the natural image domain, spatial embedding-based instance segmentation methods are known to yield high-quality results, but their utility to biomedical data is largely unexplored. (mpi-cbg.de)
  • Disease modelling using human pluripotent stem cells: Benvenisty's laboratory was the first to show a model for a human disease using human pluripotent stem cells. (wikipedia.org)
  • Benvenisty's laboratory was the first to study the immunogenicity of the cells, and to analyze the basis of their tumorigenicity. (wikipedia.org)
  • The biological properties and clinical potential of stem cells elicit that are generated must not be unduly sensitive to small fluctu- continued scientific, commercial, and public interest. (lu.se)
  • For example, mouse models of age-related neurodegenerative diseases fail to capture key features because the diseases typically strike humans in their 60s and 70s, whereas mice live for only 2 or 3 years. (nationalacademies.org)
  • A group of heterogeneous lymphoid tumors generally expressing one or more B-cell antigens or representing malignant transformations of B-lymphocytes. (lookformedical.com)
  • However, human anatomical terms do not always superimpose on the mouse, and the lack of accurate and standardised nomenclature is hampering the utility of such animal models. (atlas-d2k.org)
  • The revised ontology will be an important tool for researchers studying urogenital development/malformation in mouse models and will improve our capacity to appropriately interpret these with respect to the human situation. (atlas-d2k.org)
  • The only common feature among these tumors is the absence of giant REED-STERNBERG CELLS, a characteristic of Hodgkin's disease. (lookformedical.com)
  • BACKGROUND: During normal human kidney development, nephrogenesis (the formation of nephrons) is complete by term birth, with the majority of nephrons formed late in gestation. (atlas-d2k.org)
  • The aim of this study was to morphologically examine nephrogenesis in fetal human kidneys from 20 to 41weeks of gestation. (atlas-d2k.org)
  • The proportions of podocytes, endothelial and non-epithelial cells within mature glomeruli were stable throughout gestation. (atlas-d2k.org)
  • Lines I3 and I6 possess normal XX and a normal XY karyotype while BG01V is a variant cell line with an abnormal karyotype derived from the karyotypically normal cell line BG01. (biomedcentral.com)
  • Chapter 2 presents the science behind these models and describes the challenges of measuring and monitoring such characteristics and capacities in human neural organoids, transplants, and chimeras. (nationalacademies.org)
  • neural organoids, transplants, and chimeras, and then at issues specific to human neural transplants and chimeras or to neural organoids. (nationalacademies.org)
  • Ethical issues common to human neural organoids, transplants, and chimeras include (1) the ethical value of relieving human suffering and disease, (2) concerns about encroachment on divine roles, and (3) ethical issues related to human donors of biological materials. (nationalacademies.org)
  • Consequently, selection of PSCs with high pluripotency is essential to ensure the security and effectiveness of PSC-derived cells. (tam-receptor.com)
  • The nature of the stem cell substates and their relationship to commitment to differ- entiate and lineage selection can be elucidated in terms of a landscape picture in which stable states can be defined mathematically as attractors. (lu.se)
  • Apoptosis is the mechanism responsible for the physiological deletion of cells and appears to be intrinsically programmed. (lookformedical.com)
  • The human body is made up of about 220 different kinds of specialized cells such as nerve cells, muscle cells, fat cells and skin cells. (benthamscience.com)
  • They found the compounds activate nerve cells in the male mouse's nose with unprecedented effectiveness. (vetscite.org)
  • In a new preface, Harris offers a glimpse at the new science and technology to come, equipping readers with the knowledge to assess the ethics and policy dimensions of future forms of human enhancement. (philpapers.org)
  • Stem cell technology is the latest development in this controversial branch of science. (edu.au)
  • A molecular program that causes the interspecific barrier might be independent of genome-wide evolution. (frontiersin.org)
  • In the coming decade, we anticipate systematic enhancer perturbation studies to link non-coding variants to molecular mechanisms, changes in cell state, and disease phenotypes. (bvsalud.org)
  • The ethical and legal controversies that were aroused in the ART debates during the 1980s have been re-ignited with the development of stem cell technology. (edu.au)
  • The unique properties of human stem cells have aroused considerable optimism about their potential as new pathways for alleviating human suffering caused by disease and injury. (edu.au)
  • Our project focuses on characterizing and optimizing this approach to improve tubular maturation and cellular function and generating reporter lines for the isolation of specific cell types. (atlas-d2k.org)
  • Single-cell RNA-sequencing unveiled transcriptomic similarities between advanced human peri-gastruloids and primary peri-gastrulation cell types found in humans and non-human primates. (bvsalud.org)