Droplet digital PCR for oncogenic KMT2A fusion detection

Acute myeloid leukemia (AML) is an aggressive blood cancer diagnosed in approximately 120,000 individuals worldwide each year. During treatment for AML, detecting residual disease is essential for prognostication and treatment decision-making. Currently, methods for detecting residual AML are limited to identifying approximately 1:100 to 1:1000 leukemic cells (morphology and DNA sequencing) or are difficult to implement (flow cytometry). AML arising after chemotherapy or radiation exposure is termed therapy-related AML (t-AML) and is exceptionally aggressive and treatment resistant. t-AML is often driven by oncogenic fusions that result from prior treatments that introduce double-strand DNA breaks. The most common t-AML-associated translocations affect KMT2A. There are at least 80 known KMT2A fusion partners, but approximately 80% of fusions involve only five partners-AF9, AF6, AF4, ELL, and ENL. We present a novel droplet digital PCR assay targeting the most common KMT2A-rearrangements to enable detection of rare AML cells harboring these fusions. This assay was benchmarked in cell lines and patient samples harboring oncogenic KMT2A fusions and demonstrated a limit of detection of approximately 1:1,000,000 cells. Future application of this assay could improve disease detection and treatment decision-making for patients with t-AML with KMT2A fusions and premalignant oncogenic fusion detection in at-risk individuals after chemotherapy exposure

Connexins and Pannexins in Bone and Skeletal Muscle

PURPOSE OF REVIEW: To discuss current knowledge on the role of connexins and pannexins in the musculoskeletal system. RECENT FINDINGS: Connexins and pannexins are crucial for the development and maintenance of both bone and skeletal muscle. In bone, the presence of connexin and more recently of pannexin channels in osteoblasts, osteoclasts, and osteocytes has been described and shown to be essential for normal skeletal development and bone adaptation. In skeletal muscles, connexins and pannexins play important roles during development and regeneration through coordinated regulation of metabolic functions via cell-to-cell communication. Further, under pathological conditions, altered expression of these proteins can promote muscle atrophy and degeneration by stimulating inflammasome activity. In this review, we highlight the important roles of connexins and pannexins in the development, maintenance, and regeneration of musculoskeletal tissues, with emphasis on the mechanisms by which these molecules mediate chemical (e.g., ATP and prostaglandin E2) and physical (e.g., mechanical stimulation) stimuli that target the musculoskeletal system and their involvement in the pathophysiological changes in both genetic and acquired diseases

Psychometric Properties of the Hoarding Rating Scale-Interview

The present study tested the psychometric properties of an expanded version of the Hoarding Rating Scale (HRS-I), a semistructured interview for hoarding disorder (HD). Eighty-seven adults with HD and 44 healthy control (HC) participants were assessed using the HRS-I and completed a battery of self-report measures of HD severity, negative affect, and functional impairment. All interviews were audio recorded. From the HD participants, 21 were randomly selected for inter-rater reliability (IRR) analysis and 11 for test-retest reliability (TRR) analysis. The HRS-I showed excellent internal consistency (α = 0.87). IRR and TRR in the HD sample were good (intra-class coefficients = 0.81 and 0.85, respectively). HRS-I scores correlated strongly with scores on the self-report Saving Inventory-Revised (SI-R); partial correlations indicated that the HRS-I clutter, difficulty discarding, and acquiring items correlated significantly and at least moderately with corresponding SI-R subscales, when controlling for the other SI-R subscales. The HD group scored significantly higher on all items than did the HC group, with large effect sizes (d = 1.28–6.58). ROC analysis showed excellent sensitivity (1.00) and specificity (1.00) for distinguishing the HD and HC groups with a cutoff score of 11. Results and limitations are discussed in light of prior research

Potential Use of Morphological Characteristics in Evaluating Natural Variation of “Barako” Seedlings

The Philippines is one of only three countries able to produce Coffea liberica and grow two other species of coffee, Coffea canephora, Coffea arabica. Coffea liberica, or “Barako,” is the least cultivated of coffee species. Its unique taste has high potential to grow in the market if production was increased. However, the 2020 Taal Volcano eruption resulted in thousands of damaged Barako trees in 2020 and 2021. As new seedlings are produced, the question of variety among planting material comes up. Farmers need a method to evaluate natural differences of current Barako trees, to answer whether seedlings from farms cultivating Barako are naturally different. This research aims to determine if the morphological characteristics of cultivated seedlings from different farms can be classified through image and statistical analysis. Basic knowledge on how varieties―which producedifferent flavors, aroma, and market value of coffee―differ among farms is needed for a strategy to increase the number of seedlings. This study’s initial data set indicated statistically significant differences in the average seedling height per node and the leaf area per length of 31, 10-month old coffee seedlings from two different farms, grown in a common environment. The process may be developed further for use in evaluating natural variation among C. liberica as seedlings

Possible futures for groundwater in Burkina Faso under a changing climate

This narrative describes three possible future scenarios for water resources in Burkina Faso and the human and socio-economic impacts that might be experienced by people living in rural areas. The narratives aim to stimulate discussion towards realistic policy responses and the decision support tools needed to assist future planning needs. It is important to recognise that the scenarios do not represent every possible outcome projected by hydroclimate models, and resulting impacts will be contextualised by local circumstances. See here for information about BRAVE (www.walker.ac.uk) and the UPGro programme: upgro.org/consortium/brave2/

Spatial mapping of hematopoietic clones in human bone marrow

UNLABELLED: Clonal hematopoiesis (CH) is the expansion of somatically mutated cells in the hematopoietic compartment of individuals without hematopoietic dysfunction. Large CH clones (i.e., \u3e2% variant allele fraction) predispose to hematologic malignancy, but CH is detected at lower levels in nearly all middle-aged individuals. Prior work has extensively characterized CH in peripheral blood, but the spatial distribution of hematopoietic clones in human bone marrow is largely undescribed. To understand CH at this level, we developed a method for spatially aware somatic mutation profiling and characterized the bone marrow of a patient with polycythemia vera. We identified the complex clonal distribution of somatic mutations in the hematopoietic compartment, the restriction of somatic mutations to specific subpopulations of hematopoietic cells, and spatial constraints of these clones in the bone marrow. This proof of principle paves the way to answering fundamental questions regarding CH spatial organization and factors driving CH expansion and malignant transformation in the bone marrow. SIGNIFICANCE: CH occurs commonly in humans and can predispose to hematologic malignancy. Although well characterized in blood, it is poorly understood how clones are spatially distributed in the bone marrow. To answer this, we developed methods for spatially aware somatic mutation profiling to describe clonal heterogeneity in human bone marrow. See related commentary by Austin and Aifantis, p. 139

Effects of immunization against bone morphogenetic protein-15 and growth differentiation factor-9 on ovarian function in mares

Currently there is no contraceptive vaccine that can cause permanent sterility in mares. This study investigates the effect of vaccination against oocyte-specific growth factors, Bone Morphogenetic Protein 15 (BMP-15) and Growth Differentiation Factor 9 (GDF-9), on ovarian function of mares. It was hypothesized that immunization against these growth factors would prevent ovulation and/or accelerate depletion of the oocyte reserve. For this study, 30 mares were randomly assigned to three groups (n=10/group) and vaccinated with BMP-15 or GDF-9 peptides conjugated to KLH and adjuvant, or a control of phosphate buffered saline and adjuvant. Horses received vaccinations at weeks 0, 6, 12, and 18. Ovarian activity and estrous behavior were evaluated 3 days a week via ultrasonography and interaction with a stallion. The study was initiated on March1, 2016. Upon evaluation of ovulation rate, the GDF-9 group did not have a difference (P=0.66) in ovulation rate when compared to controls (10.8 and 10.0 ovulations, respectively), but the number of ovulations in the BMP-15 group was less (P=0.02; 4.9 ovulations). Average follicle size prior to ovulation was less (P \u3c 0.0001) in both treatment groups compared to controls. Estrous behavior was altered in both the BMP-15 and GDF-9 groups compared to controls after the second vaccination (P=0.05 and 0.03, respectively). Although further research is required to determine the continued effects of vaccination against GDF-9 on ovulation rates, these results indicate that vaccination against BMP-15 and GDF-9 could serve as a contraceptive in wild horse populations

Defective cancellous bone structure and abnormal response to PTH in cortical bone of mice lacking Cx43 cytoplasmic C-terminus domain

Connexin 43 (Cx43) forms gap junction channels and hemichannels that allow the communication among osteocytes, osteoblasts, and osteoclasts. Cx43 carboxy-terminal (CT) domain regulates channel opening and intracellular signaling by acting as a scaffold for structural and signaling proteins. To determine the role of Cx43 CT domain in bone, mice in which one allele of full length Cx43 was replaced by a mutant lacking the CT domain (Cx43(ΔCT/fl)) were studied. Cx43(ΔCT/fl) mice exhibit lower cancellous bone volume but higher cortical thickness than Cx43(fl/fl) controls, indicating that the CT domain is involved in normal cancellous bone gain but opposes cortical bone acquisition. Further, Cx43(ΔCT) is able to exert the functions of full length osteocytic Cx43 on cortical bone geometry and mechanical properties, demonstrating that domains other than the CT are responsible for Cx43 function in cortical bone. In addition, parathyroid hormone (PTH) failed to increase endocortical bone formation or energy to failure, a mechanical property that indicates resistance to fracture, in cortical bone in Cx43(ΔCT) mice with or without osteocytic full length Cx43. On the other hand, bone mass and bone formation markers were increased by the hormone in all mouse models, regardless of whether full length or Cx43(ΔCT) were or not expressed. We conclude that Cx43 CT domain is involved in proper bone acquisition; and that Cx43 expression in osteocytes is dispensable for some but not all PTH anabolic actions