Bone Marrow Wars: Attack of the Clones

Multiple myeloma is characterized by the malignant proliferation of clonal plasma cells producing monoclonal paraproteins, leading to multi-organ damage. On the other hand monoclonal B-cell lymphocytosis (MBCL) is characterized by the malignant proliferation of clonal B-lymphocytes, with potential to develop into chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL). CLL/SLL can result in visceromegaly, anemia, thrombocytopenia, fevers, night sweats and unintentional weight loss. Literature review demonstrates these two malignant clonal bone marrow disorders are most frequently seen independently in patients; however, we report one rare diagnostic challenge where both clonal disorders were identified in a single patient concurrently. A 64-year-old man initially presented with worsening back pain. Thoracic spine x-ray revealed a T11 compression fracture, confirmed by magnetic resonance imaging. Complete blood count revealed a white blood cell count of 7.3 K/uL with 54% lymphocyte predominance and peripheral smear demonstrated a population of small lymphocytes with round nuclei and an atypical chromatin pattern suggestive of CLL/MBCL. Flow cytometry revealed a monoclonal B-cell CD5 positive, CD23 positive, CD10 negative population with an absolute count of 1.6 K/uL. Due to the instability and pain associated with the spinal fracture, patient had kyphoplasty performed and intraoperative bone biopsies were taken from both T11 and T12 vertebrae. Interestingly each bone biopsy revealed involvement by both a kappa-light chain restricted plasma cell neoplasm, ranging from 15% to 30% cellularity, as well as a CD5-positive B-cell lymphocyte population. It suggested two concurrent but pathologically distinct pathologies including plasma cell myeloma and a separate B-cell lymphoproliferative disorder with immunophenotypic features suggestive of CLL/MBCL. Bone marrow biopsy was performed for definitive evaluation and confirmed multiple myeloma with 15-20% kappa-restricted plasma cells identified, and also confirmed concurrent MBCL with CD5 and CD23-positive, kappa-restricted B-cells identified on bone marrow flow cytometry. Adding an additional layer of complexity, bone marrow molecular genetics revealed presence of a MYD88 mutation, raising concern for possible lymphoplasmacytic lymphoma (LPL). However, secondary pathologic review ruled out LPL, as the immunophenotypic pattern of the clonal B-cells was not consistent with that of LPL, and although the MYD88 mutation is predominantly seen in LPL, it has also been seen in a small percentage of CLL/SLL cases and exceedingly rarely described in MM as well. Serum protein electrophoresis with immunofixation, serum quantitative immunoglobulins and serum quantitative free light chain assay revealed findings consistent with IgG kappa multiple myeloma and systemic CT imaging was negative for any lymphadenopathy, confirming MBCL. Patient was started on first-line multiple myeloma systemic therapy for transplant eligible patients and has demonstrated an excellent response to treatment thus far. This patient case serves to demonstrate the importance of maintaining a broad differential when approaching hematological problems; It also underlines the necessity for a complete diagnostic evaluation to identify rare clinical conundrums such as with our patient, allowing for proper and timely treatment. While we use “Occam’s razor” to explain multiple problems with a single unifying diagnosis the rare possibility of divergent diagnosis is to be always entertained

Attack of the Clones: Legislative Approaches to Human Cloning in the United States

The legal concerns involving the application of cloning technology to humans should be of utmost concern, as the area is extremely complex. Cloning could potentially have great benefits or disastrous effects. Lawmakers have been careful to make certain that the legislation passed is comprehensive and useful for regulation of the ever-changing field of cloning. From debates on whether reproductive or therapeutic cloning should be permitted or banned, to concerns as to who has jurisdiction over cloning, the battle to develop cloning legislation has been difficult. However, this iBrief argues that the currently-proposed federal legislation is constitutional

Attack of the Clones: Legislative Approaches to Human Cloning in the United States

The legal concerns involving the application of cloning technology to humans should be of utmost concern, as the area is extremely complex. Cloning could potentially have great benefits or disastrous effects. Lawmakers have been careful to make certain that the legislation passed is comprehensive and useful for regulation of the ever-changing field of cloning. From debates on whether reproductive or therapeutic cloning should be permitted or banned, to concerns as to who has jurisdiction over cloning, the battle to develop cloning legislation has been difficult. However, this iBrief argues that the currently-proposed federal legislation is constitutional

23andMe: Attack of the Clones and Other Concerns

A few years ago, ancestry websites took the world by storm. People were fascinated with their history and heritage and wanted to find out more about where they came from. Then along came 23andMe, which allowed people to not only unearth their familial roots, but also bring to light unknown medical conditions or predispositions to certain medical issues. 23andMe then took the unprecedented step of teaming up with a pharmaceutical company to create drugs with its users’ genetic information. After this announcement, some users were caught off guard, having had no idea that their genetic information—something so sensitive and uniquely personal to them—was being used to create drugs. While 23andMe presented this possibility in their Research Consent Document, it is clear that many users either did not read it or simply did not understand the terms of their participation. This begs the question: how do users effectively pull their genetic information from research they did not necessarily intend to participate in? Neither the current American statutory scheme nor property and contract case law provide these users with protection or any way to withdraw from all research they deem unacceptable. Courts have ruled that people who allow their genetic information to be used for research forfeit their property rights to it and that it is not relevant if people did not read the consent form they agreed to, as long as they were put on notice of additional terms. Since these avenues for legal recourse are essentially blocked for users that want to reclaim their genetic information, they should instead pursue a clearer path. Examining gametic material jurisprudence— a similarly situated but more consumer-friendly area of law which involves disputes over the rights to the genetic information found in eggs, sperm, and embryos—may just reveal that new path. This Note examines the various issues 23andMe’s research program presents for users who wish to fully remove their genetic information from 23andMe’s research given the current American statutory scheme and case law in various American jurisdictions. Under these legal frameworks, the courts do not look to the intent of the parties in deciding who has rights to the genetic material. Rather, courts look to whether there was a forfeiture, consent, and notice of terms. This is in spite of the well-documented fact that people often do not fully read or understand contracts, especially internet contracts, when they agree to them. In contrast, gametic material jurisprudence looks beyond contracts and certain acts to the intent of the donors in deciding who has rights to the gametic material. This legal framework recognizes the reality that people do not necessarily read or understand what they agree to when they allow their genetic material to be used in research and gives those who did not intend to participate in certain kinds of research a way to permanently reclaim their genetic material

Attack Of The Clones: Copyright Protection For Video Game Developers

This comment focuses on the case Tetris Holding, LLC v. Xio Interactive, Inc. and uses it to explore how video game developers’ original innovations are protected under copyright law so as to fully restrict those wishing to “clone” these innovations for their own financial gain. The comment begins by outlining relevant copyright concepts and statutes, using case examples, and detailing the analytical framework courts use to evaluate claims asserted under copyright law. It then discusses the accommodations by both the video game industry as well as online marketplaces in order to protect original content from being cloned. After evaluating these existing accommodations, the comment outlines the financial and intellectual effect full compliance under copyright law would have on the video game industry. Lastly, the comment suggests possible action for courts and those within the video game industry to take in order to protect original ideas by developers, ultimately concluding that games on all platforms be awarded copyright certification before their release

Attack of the clones: an investigation into removing redundant source code

Long-term maintenance of code will often lead to the introduction of duplicated or 'cloned' code. Legacy systems riddled with these clones have large amounts of redundant code and are more difficult to understand and maintain. One option available to improve maintainability and to increase software reuse, is to re-engineer code clones into reusable components. However, before this can be achieved detection and removal of this redundant code is necessary. There are several established clone detection tools for software maintenance and this thesis aims to investigate the similarities between their output. It also looks at how maintainers may best use them to reduce the amount of redundant code in a software system. This will be achieved by running clone detection tools on several different case studies. Included in these case studies will be a novel tool called Covet inspired by research of Mayrand [May96b] which attempted to identify cloned routines through a comparison of software metrics generated from each routine. It was found that none of the clone detection tools achieved either 100% precision or 100% recall. Each tool identified very different sets of clones. Overall MOSS achieved the greatest precision and CCFinder the greatest recall. Also observed was that the use of automatically generated code increased the proportion of clones found in a software system

Attack of the clones: microglia in health and disease

[INTRODUCTION] Microglia are brain-resident macrophages that carry out immune surveillance, support neurogenesis and neuronal survival, shape the neuronal network, and maintain tissue homeostasis (Nimmerjahn et al., 2005; Hanisch and Kettenmann, 2007; Sierra et al., 2010; Tremblay et al., 2010; Schafer et al., 2012; Ueno et al., 2013; Squarzoni et al., 2014; Schafer and Stevens, 2015; Diaz-Aparicio et al., 2020). The adult resident pool of microglia is primarily derived from yolk sac (YS) erythromyeloid progenitors (EMPs) that have clonally proliferated within the brain parenchyma during development (Alliot et al., 1999; Ginhoux et al., 2010; Hashimoto et al., 2013; Gomez Perdiguero et al., 2015). To maintain their cell density in adulthood, the resident microglial cells undergo local clonal self-renewal (Ajami et al., 2007; Askew et al., 2017; Réu et al., 2017; Tay et al., 2017). A study on parabiotic chimeric mice revealed that the resident microglial population is exclusively replenished by locally derived microglial clones with no evidence of contribution from peripheral myeloid progenitors (Ajami et al., 2007). Clonal expansion of non-ablated residual microglia was also found to be responsible for re-establishing steady state microglial cell densities following a pharmacological ablation (Huang et al., 2018). Homeostatic microglia continuously monitor the brain environment, scavenge dying cells and cellular debris, and rapidly respond to any tissue damage (Kreutzberg, 1996; Streit et al., 1999). In response to acute pathology, resident microglia rapidly accumulate around the lesion via clonal microgliosis (Streit et al., 1999; Ladeby et al., 2005; Ajami et al., 2007; Ransohoff, 2007). Microgliosis at the site of CNS damage has been shown to be governed by signaling molecules like colony-stimulating factor-1 (CSF1), fractalkine receptor (CX3CR1) and purinergic receptor P2Y12 (P2RY12) (Guan et al., 2016; Gu et al., 2016; Peng et al., 2016). To repair damage, microglia elicit proinflammatory cytokines and later transition to anti-inflammatory phenotypes (Colton, 2009; Lloyd et al., 2019). Several studies have suggested that clinical recovery of acute lesions is accompanied by the resolution of proliferated microglia by migration and cell death (Dihné et al., 2001; Wilson et al., 2004; Tay et al., 2017; Lloyd et al., 2019). However, in severe or chronic neurodegenerative pathologies like Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS), and motor neuron diseases, microglial clonal expansion has been persistently observed around lesions and plaques (Glass et al., 2010; Streit et al., 2020). In AD and MS animal models, microglial proliferation around the Aβ plaques and demyelinating neurons was actively promoted by CSF1R and triggering receptor expressed on myeloid cells 2 (TREM2) (Cantoni et al., 2015; Olmos-Alonso et al., 2016; Wang et al., 2016; Jay et al., 2017; Gushchina et al., 2018; Zhao et al., 2018). CSF1R, transforming growth factor beta (TGFβ), and purinergic signaling pathways that influence microglial cell densities are impaired in neurodegenerative diseases (Gómez-Nicola et al., 2013; Von Bernhardi et al., 2015; Olmos-Alonso et al., 2016; Pietrowski et al., 2021). Taken together, the capacity for microglial clonal expansion, proliferation, or renewal, clearly play an important function across CNS development, health, and disease. Although clonal expansion and renewal of microglia appears necessary for physiological brain development, maintenance of CNS health, and response to acute damage, whether microglial clones are beneficial or detrimental in chronic neurodegeneration remains unclear. In this opinion article, we discuss the implications of the formation of microglial clones in health and disease, independent of peripheral myeloid recruitment to the CNS in similar contexts (Figure 1). Several studies have claimed that microglia surrounding plaques and lesions exert detrimental effects and exacerbate disease conditions (Streit et al., 2009; Lassmann et al., 2012; Keren-Shaul et al., 2017; Krasemann et al., 2017; Shahidehpour et al., 2021). Considering that the restoration of tissue homeostasis coincides with the resolution of microglial clones to regain steady state microglial tiling (Dihné et al., 2001; Wilson et al., 2004; Tay et al., 2017; Lloyd et al., 2019), we hypothesize that unresolved microglial clones contribute to the prolongation of neurodegenerative states in chronic neuropathologies. As a hallmark of CNS pathology, microgliosis is likely important to limit tissue damage and infection and for local repair, as is typical in inflammatory responses of tissue-resident macrophages (Jenkins et al., 2011). A timely resolution of excess microglia resulting from clonal expansion is expected to aid or accompany the restoration of homeostasis and clinical recovery. However, sustained presence of reactive microglial clones at high densities around lesions or plaques with no signs of resolution likely lead to neurotoxic outcomes. To explore our hypothesis, we examine various contexts in which microglial clonal expansion has taken place and consider if strategic targeting of microglial clones could ameliorate chronic disease states.https://www.frontiersin.org/articles/10.3389/fncel.2022.831747/ful

ATTACK OF THE CLONES: ELUCIDATING THE ROLE OF CLONALITY IN THE INVASION SUCCESS OF CARPOBROTUS EDULIS

Transcriptomics is a modern technique in genomics that utilizes RNA sequences to get a snapshot of genetic expression. This is a powerful tool in non-model species lacking a reference genome. Thus, the application of comparative transcriptomics has the potential to help us elucidate the evolutionary mechanisms that facilitate species invasion. Carpobrotus edulis is a prolific and widespread invasive succulent plant belonging to the Aizoaceae family. A native to South Africa, this species has become a dominant invader of many Mediterranean coastal areas. In this study, we leveraged the use of RNAseq to investigate evolutionary changes among invasive populations. RNA-seq data was gathered from experimental native and invasive populations tested under stress conditions and used to assemble a de novo transcriptome and perform a differential expression analysis. We provide the first annotated transcriptome for the plant genus Carpobrotus and the Aizoaceae family. Differential expression analysis identified several differentially expressed transcripts between native and invasive populations. Invasive populations showed no increases in putative genes related to flowering and decreased expression in genes related to protection and defense. Supporting the hypothesis that C. edulis has undergone significant genetic modification in the invaded range. These results are expected under the Evolution of Increased Competitive Ability hypothesis and Genetic Accommodation hypothesis, wherein novel metabolic processes and changes in gene regulatory networks evolve when introduced to ecologies