Exit of pediatric pre-B acute lymphoblastic leukaemia cells from the bone marrow to the peripheral blood is not associated with cell maturation or alterations in gene expression

<p>Abstract</p> <p>Background</p> <p>Childhood pre-B acute lymphoblastic leukemia (ALL) is a bone marrow (BM) derived disease, which often disseminates out of the BM cavity, where malignant cells to a variable degree can be found circulating in the peripheral blood (PB). Normal pre-B cells are absolutely dependent on BM stroma for survival and differentiation. It is not known whether transformed pre-B ALL cells retain any of this dependence, which possibly could impact on drug sensitivity or MRD measurements.</p> <p>Results</p> <p>Pre-B ALL cells, highly purified by a novel method using surface expression of CD19 and immunoglobulin light chains, from BM and PB show a very high degree of similarity in gene expression patterns, with differential expression of vascular endothelial growth factor (VEGF) as a notable exception. In addition, the cell sorting procedure revealed that in 2 out of five investigated patients, a significant fraction of the malignant cells had matured beyond the pre-B cell stage.</p> <p>Conclusion</p> <p>The transition of ALL cells from the BM into the circulation does not demand, or result in, major changes of gene expression pattern. This might indicate an independence of BM stroma on the part of transformed pre-B cells, which contrasts with that of their normal counterparts.</p

Vitamin D Induction of the Human Antimicrobial Peptide Cathelicidin in the Urinary Bladder

The urinary tract is frequently being exposed to potential pathogens and rapid defence mechanisms are therefore needed. Cathelicidin, a human antimicrobial peptide is expressed and secreted by bladder epithelial cells and protects the urinary tract from infection. Here we show that vitamin D can induce cathelicidin in the urinary bladder. We analyzed bladder tissue from postmenopausal women for expression of cathelicidin, before and after a three-month period of supplementation with 25-hydroxyvitamin D3 (25D3). Cell culture experiments were performed to elucidate the mechanisms for cathelicidin induction. We observed that, vitamin D per se did not up-regulate cathelicidin in serum or in bladder tissue of the women in this study. However, when the bladder biopsies were infected with uropathogenic E. coli (UPEC), a significant increase in cathelicidin expression was observed after 25D3 supplementation. This observation was confirmed in human bladder cell lines, even though here, cathelicidin induction occurred irrespectively of infection. Vitamin D treated bladder cells exerted an increased antibacterial effect against UPEC and colocalization to cathelicidin indicated the relevance of this peptide. In the light of the rapidly growing problem of resistance to common urinary tract antibiotics, we suggest that vitamin D may be a potential complement in the prevention of UTI

ENIGMA-anxiety working group : Rationale for and organization of large-scale neuroimaging studies of anxiety disorders

Altres ajuts: Anxiety Disorders Research Network European College of Neuropsychopharmacology; Claude Leon Postdoctoral Fellowship; Deutsche Forschungsgemeinschaft (DFG, German Research Foundation, 44541416-TRR58); EU7th Frame Work Marie Curie Actions International Staff Exchange Scheme grant 'European and South African Research Network in Anxiety Disorders' (EUSARNAD); Geestkracht programme of the Netherlands Organization for Health Research and Development (ZonMw, 10-000-1002); Intramural Research Training Award (IRTA) program within the National Institute of Mental Health under the Intramural Research Program (NIMH-IRP, MH002781); National Institute of Mental Health under the Intramural Research Program (NIMH-IRP, ZIA-MH-002782); SA Medical Research Council; U.S. National Institutes of Health grants (P01 AG026572, P01 AG055367, P41 EB015922, R01 AG060610, R56 AG058854, RF1 AG051710, U54 EB020403).Anxiety disorders are highly prevalent and disabling but seem particularly tractable to investigation with translational neuroscience methodologies. Neuroimaging has informed our understanding of the neurobiology of anxiety disorders, but research has been limited by small sample sizes and low statistical power, as well as heterogenous imaging methodology. The ENIGMA-Anxiety Working Group has brought together researchers from around the world, in a harmonized and coordinated effort to address these challenges and generate more robust and reproducible findings. This paper elaborates on the concepts and methods informing the work of the working group to date, and describes the initial approach of the four subgroups studying generalized anxiety disorder, panic disorder, social anxiety disorder, and specific phobia. At present, the ENIGMA-Anxiety database contains information about more than 100 unique samples, from 16 countries and 59 institutes. Future directions include examining additional imaging modalities, integrating imaging and genetic data, and collaborating with other ENIGMA working groups. The ENIGMA consortium creates synergy at the intersection of global mental health and clinical neuroscience, and the ENIGMA-Anxiety Working Group extends the promise of this approach to neuroimaging research on anxiety disorders

Oral rehydration therapies in Senegal, Mali, and Sierra Leone: a spatial analysis of changes over time and implications for policy.

BACKGROUND: Oral rehydration solution (ORS) is a simple intervention that can prevent childhood deaths from severe diarrhea and dehydration. In a previous study, we mapped the use of ORS treatment subnationally and found that ORS coverage increased over time, while the use of home-made alternatives or recommended home fluids (RHF) decreased, in many countries. These patterns were particularly striking within Senegal, Mali, and Sierra Leone. It was unclear, however, whether ORS replaced RHF in these locations or if children were left untreated, and if these patterns were associated with health policy changes. METHODS: We used a Bayesian geostatistical model and data from household surveys to map the percentage of children with diarrhea that received (1) any ORS, (2) only RHF, or (3) no oral rehydration treatment between 2000 and 2018. This approach allowed examination of whether RHF was replaced with ORS before and after interventions, policies, and external events that may have impacted healthcare access. RESULTS: We found that RHF was replaced with ORS in most Sierra Leone districts, except those most impacted by the Ebola outbreak. In addition, RHF was replaced in northern but not in southern Mali, and RHF was not replaced anywhere in Senegal. In Senegal, there was no statistical evidence that a national policy promoting ORS use was associated with increases in coverage. In Sierra Leone, ORS coverage increased following a national policy change that abolished health costs for children. CONCLUSIONS: Children in parts of Mali and Senegal have been left behind during ORS scale-up. Improved messaging on effective diarrhea treatment and/or increased ORS access such as through reducing treatment costs may be needed to prevent child deaths in these areas

To be or not to be, unraveling molecular mechanisms for lineage decisions in developing blood cells

All hematopoietic cells originate from hematopoietic stem cells (HSC) residing in the BM. The process of differentiation, through which HSCs generate progenitors and subsequently mature blood cells has been extensively studied but many of the regulatory mechanisms involved remains elusive. The classical model for hematopoiesis has been established based on the identification of common myeloid precursors and common lymphoid precursors, suggesting that the first lineage commitment step results in the strict separation between the lymphoid and myeloid lineages. However, by subfractionation of the LIN-SCA1+KIT+ bone marrow population, containing all HSCs in the mouse, based on expression of CD34 and FLT3, three functionally distinct populations were identified. CD34- cells contained the HSCs, CD34+FLT3- short term repopulating cells and CD34+ FLT3+/hi cells with combined lymhpoid and GM potential but, only a small fraction (3%) displaying MkE potential in vitro. In agreement with this MkE associated genes (Gata1, Epor, Vwf etc) were found to be down-regulated in CD34+FLT3hi cells while lymphoid associated genes (Il7r, Rag1, Tdt, sterile IgH transcripts) were found to be up-regulated both at the population and single cell level. Based on this we proposed a new model for hematopoietic development were the first restriction point involves the separation of MkE and lymphoid potential while retaining GM potential. In order to investigate the continued development of B-lymphoid cells we investigated the functional role of the transcription factor EBF1, known to be of critical importance for B cell development. By means of comparativ and correlativ analysis of microarray data from cell-lines we, in addition to the previously known EBF1 target genes, identified Vpreb3, CD19, Ceacam1 and CD53 as EBF1 target genes. Furthermore, by transplantations of EBF1 deficient fetal liver cells, we were able to show EBF1 to be dispensable for the generation of CLPs but that lack of EBF1 expression resulted in diminished IgH DJ recombination and reduced expression of B-lineage associated genes already at this early stage of development. Using a hCD25 lambda5-promoter transgene as a marker for EBF1 activity a small CLP subpopulation (~6%) could be identified. This population demonstrated diminished T-cell potential, increased B-cell potential, as well as up-regulation of Vpreb, lambda5, B29, Mb1 and importantly Pax5. Taken together this suggest that B-lineage commitment occurs already at the level of the CLP

The legality of humanitarian intervention in customary international law

The aim of this study is to examine whether there exists a right of humanitarian intervention in customary international law. The point of departure is the principle of non-use of force in customary international law. The International Court of Justice established in the Nicaragua Case that the principle of non-use of force not only is a jus cogens norm, but that it also has the basic identity of the UN-charter article 2(4). By interpreting article 2(4) according to the provisions of interpretation in the Vienna Convention on the Law of Treaties, I thereby conclude that humanitarian intervention is not in conformity with article 2(4) and consequently not in conformity with the principle of non-use of force either. The high threshold for modification of the principle of non-use of force due to its jus cogens character is furthermore established. It is submitted that the evidence of State practice for modification of the principle of non-use of force must be overwhelming and reflect the legal conviction of the international community at large. I then conduct a case study, where four cases, which could be perceived as evidence of State practice in favor of humanitarian intervention, are analyzed. The cases are ''the three best cases'' during the Cold War: the Indian invasion of East Pakistan 1971, the Vietnamese invasion of Kampuchea 1978-79, the Tanzanian invasion of Uganda 1978-1979, as well as Operation Allied Force conducted by NATO against the FRY in the spring of 1999. The invading states in ''the three best cases'' primarily relied on self-defense as a way of arguing the legality of the interventions, which reinforces the principle of non-use of force in customary international law. The reaction from the international community was overall negative while the few states, which approved of the invasions, did not rely on the concept of humanitarian intervention. Operation Allied Force indicated a change in the attitude towards humanitarian intervention. The majority of the NATO-states together with the international community chose to assert the morality of the operation however. It has little value in terms of establishing a right of humanitarian intervention in customary international law. I therefore conclude that the State practice analyzed can not be used as evidence for a right of humanitarian intervention in customary international law, especially since the principle of non-use of force is a jus cogens norm. Regarding the question of humanitarian intervention de lege ferenda, I am of the opinion that humanitarian intervention should not be part of international law, because of the risk of abuse, which is worsened by the fact that humanitarian intervention is only available to nations with the military and economic capacity. The fact that humanitarian intervention is illegal under international law must not however stop the international community from reacting when there is a desperate humanitarian need

Regulation of gene expression in the vascular wall

Blood vessel growth and function are closely related to a number of pathological conditions, including tumor angiogenesis, wound healing and atherosclerosis. Smooth muscle cells (SMC) and endothelial cells (EC), the two major constituents of the vascular wall, are both characterized by the expression of unique phenotypic marker genes, many of which have vital roles in blood vessel development and disease. We therefore sought to obtain a more complete picture of vascular-specific gene expression, gene regulation and genetic variation. We performed an unbiased computational screen to identify cases of transcriptional coregulation in mammalian cell differentiation. This generated a number of novel hypotheses, one of them being that the SMC marker gene lipoma-preferred partner (LPP) could be activated by serum response factor (SRF), a known master regulator of SMC differentiation. Using chromatin immunoprecipitation, gel shift assays, reporter assays and transgenic mouse models, we showed that LPP belongs to the category of SMC-specific genes that are regulated by SRF, an important insight because LPP has a role in the control of SMC migration. By combining in-house and public genome-wide expression data, we identified 32 novel EC-specific mRNAs. A number of these, such as the G-protein coupled receptors Gpr116 and Ramp2, represent putative drug targets. By integrating our results with data from published genome-wide association studies, we investigated if genetic variation in EC-specific genes contributes to human disease. Independent replication of selected SNPs in 10,505 individuals revealed that a variant in one of the novel EC markers, DRAM, is associated with the development of essential hypertension. Finally, the role of microRNAs (miRNA), an abundant class of small regulatory RNAs, was evaluated in the microvasculature. Through screening of public expression data, we identified a novel microvascular-enriched miRNA, miR-145, and showed that overexpression of this molecule leads to reduced cell migration. In conclusion, we identified novel vascular marker genes and provided insights into the regulation of such genes. In addition, we showed that genetic variation in a novel EC marker gene contributes to the development of hypertension in the human population

High-throughput ChIPmentation: freely scalable, single day ChIPseq data generation from very low cell-numbers

BackgroundChromatin immunoprecipitation coupled to sequencing (ChIP-seq) is widely used to map histone modifications and transcription factor binding on a genome-wide level.ResultsWe present high-throughput ChIPmentation (HT-ChIPmentation) that eliminates the need for DNA purification prior to library amplification and reduces reverse-crosslinking time from hours to minutes.ConclusionsThe resulting workflow is easily established, extremely rapid, and compatible with requirements for very low numbers of FACS sorted cells, high-throughput applications and single day data generation