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The identification of the genes associated with chromosomal translocation breakpoints has fundamentally changed understanding of the molecular basis of hematological malignancies. By contrast, the study of chromosomal deletions has been hampered by the large number of genes deleted and the complexity of their analysis. We report the generation of a mouse model for human 5q-syndrome using large-scale chromosomal engineering. Haploinsufficiency of the Cd74-Nid67 interval (containing Rps14, encoding the ribosomal protein S14) caused macrocytic anemia, prominent erythroid dysplasia and monolobulated megakaryocytes in the bone marrow. These effects were associated with defective bone marrow progenitor development, the appearance of bone marrow cells expressing high amounts of the tumor suppressor p53 and increased bone marrow cell apoptosis. Notably, intercrossing with p53-deficient mice completely rescued the progenitor cell defect, restoring common myeloid progenitor and megakaryocytic-erythroid progenitor, granulocyte-monocyte progenitor and hematopoietic stem cell bone marrow populations. This mouse model suggests that a p53-dependent mechanism underlies the pathophysiology of the 5q-syndrome., 5q-syndrome was first described in 1974 as a consistent association of the deletion of the long arm of chromosome 5 (del(5q)) with hematological abnormalities: macrocytosis, anemia, normal or high platelet [...]

Objectives: Bioenergetic measurements in peripheral blood mononuclear cells (PBMCs) using high-throughput respirometry is a promising minimally invasive approach to studying mitochondrial function in humans. However, optimal methods for collecting PBMCs are not well studied., Methods: Bioenergetics and viability were measured across processing delays, tube type and cryopreservation., Results: Storage of collection tubes on dry ice resulted in unrecoverable samples and using the Cell Preparation Tube (CPT TM ) significantly reduced viability. Thus, storage in Sodium Citrate (NaC) and ethylenediaminetetraacetic acid (EDTA) tubes were studied in detail. Cell viability decreased by 0.5% for each hour the samples remained on wet ice prior to processing while cryopreservation decreased viability by 9.6% with viability remaining stable for about one month in liquid nitrogen. Adenosine triphosphate linked respiration (ALR) and proton-leak respiration (PLR) changed minimally while maximal respiratory capacity (MRC) and reserve capacity (RC) decreased markedly with collection tubes stored on wet ice over 24 hrs. Changes in respiratory parameters were more modest over the first 8 hours. Manipulations to replace media did not attenuate changes in respiratory parameters. Cryopreservation decreased ALR, MRC and RC by 17.20, 95.30 and 54.92 pmol/min, respectively and increased PLR by 2.65 pmol/min. PLR, MRC and RC changed moderately during the first month in liquid nitrogen for freshly frozen PBMCs., Conclusions: Our results suggest that bioenergetics in PBMCs vary based on the processing time from specimen collection and preservation method. Changes in bioenergetics can be minimized by processing samples with a minimal time delay. Changes in viability are minimal and may not correspond to changes in bioenergetics., Competing Interests: None., (AJTR Copyright © 2022.)