Vocal tract resonances in singing: variation with laryngeal mechanism for male operatic singers in chest and falsetto registers

International audienceSeven male operatic singers sang the same notes and vowels in their chest and their falsetto registers, covering the overlap frequency range where two main laryngeal mechanisms can be identified by means of electroglottography: M1 in chest register and M2 in falsetto register. Glottal contact quotients determined using electroglottography were typically lower by 0.27 in M2 than in M1. Vocal tract resonance frequencies were measured by using broadband excitation at the lips and found to be typically lower in M2 than in M1 sung at the same pitch and vowel; R1 typically by 65 Hz and R2 by 90 Hz. These shifts in tract resonances were only weakly correlated with the changes in the contact quotient or laryngeal height that were measured simultaneously. There was considerable variability in the resonance tuning strategies used by the singers, and no evidence of a uniform systematic tuning strategy used by all singers. A simple model estimates that the shifts in resonance frequencies are consistent with the effective glottal area in falsetto register (M2) being 60%-70% of its value in chest register (M1)

Therapeutic Neonatal Hepatic Gene Therapy in Mucopolysaccharidosis VII Dogs

Dogs with mucopolysaccharidosis VII (MPS VII) were injected intravenously at 2–3 days of age with a retroviral vector (RV) expressing canine β-glucuronidase (cGUSB). Five animals received RV alone, and two dogs received hepatocyte growth factor (HGF) before RV in an attempt to increase transduction efficiency. Transduced hepatocytes expanded clonally during normal liver growth and secreted enzyme with mannose 6-phosphate. Serum GUSB activity was stable for up to 14 months at normal levels for the RV-treated dogs, and for 17 months at 67-fold normal for the HGF/RV-treated dog. GUSB activity in other organs was 1.5–60% of normal at 6 months for two RV-treated dogs, which was likely because of uptake of enzyme from blood by the mannose 6-phosphate receptor. The body weights of untreated MPS VII dogs are 50% of normal at 6 months. MPS VII dogs cannot walk or stand after 6 months, and progressively develop eye and heart disease. RV- and HGF/RV-treated MPS VII dogs achieved 87% and 84% of normal body weight, respectively. Treated animals could run at all times of evaluation for 6–17 months because of improvements in bone and joint abnormalities, and had little or no corneal clouding and no mitral valve thickening. Despite higher GUSB expression, the clinical improvements in the HGF/RV-treated dog were similar to those in the RV-treated animals. This is the first successful application of gene therapy in preventing the clinical manifestations of a lysosomal storage disease in a large animal

Development and validation of an index of musculoskeletal functional limitations

BACKGROUND: While musculoskeletal problems are leading sources of disability, there has been little research on measuring the number of functionally limiting musculoskeletal problems for use as predictor of outcome in studies of chronic disease. This paper reports on the development and preliminary validation of a self administered musculoskeletal functional limitations index. METHODS: We developed a summary musculoskeletal functional limitations index based upon a six-item self administered questionnaire in which subjects indicate whether they are limited a lot, a little or not at all because of problems in six anatomic regions (knees, hips, ankles and feet, back, neck, upper extremities). Responses are summed into an index score. The index was completed by a sample of total knee replacement recipients from four US states. Our analyses examined convergent validity at the item and at the index level as well as discriminant validity and the independence of the index from other correlates of quality of life. RESULTS: 782 subjects completed all items of the musculoskeletal functional limitations index and were included in the analyses. The mean age of the sample was 75 years and 64% were female. The index demonstrated anticipated associations with self-reported quality of life, activities of daily living, WOMAC functional status score, use of walking support, frequency of usual exercise, frequency of falls and dependence upon another person for assistance with chores. The index was strongly and independently associated with self-reported overall health. CONCLUSION: The self-reported musculoskeletal functional limitations index appears to be a valid measure of musculoskeletal functional limitations, in the aspects of validity assessed in this study. It is useful for outcome studies following TKR and shows promise as a covariate in studies of chronic disease outcomes.National Institutes of Health (NIH P60 AR 47782; NIH K24 AR 02123

Carbon Capture and Water Emissions Treatment System (CCWESTRS) at Fossil-Fueled Electric Generating Plants

The Tennessee Valley Authority (TVA), the Electric Power Research Institute (EPRI), and the Department of Energy-National Energy Technologies Laboratory (DOE-NETL) are evaluating and demonstrating integration of terrestrial carbon sequestration techniques at a coal-fired electric power plant through the use of Flue Gas Desulfurization (FGD) system gypsum as a soil amendment and mulch, and coal fly ash pond process water for periodic irrigation. From January to March 2002, the Project Team initiated the construction of a 40 ha Carbon Capture and Water Emissions Treatment System (CCWESTRS) near TVA's Paradise Fossil Plant on marginally reclaimed surface coal mine lands in Kentucky. The CCWESTRS is growing commercial grade trees and cover crops and is expected to sequester 1.5-2.0 MT/ha carbon per year over a 20-year period. The concept could be used to meet a portion of the timber industry's needs while simultaneously sequestering carbon in lands which would otherwise remain non-productive. The CCWESTRS includes a constructed wetland to enhance the ability to sequester carbon and to remove any nutrients and metals present in the coal fly ash process water runoff. The CCWESTRS project is a cooperative effort between TVA, EPRI, and DOE-NETL, with a total budget of $1,574,000. The proposed demonstration project began in October 2000 and has continued through December 2005. Additional funding is being sought in order to extend the project. The primary goal of the project is to determine if integrating power plant processes with carbon sequestration techniques will enhance carbon sequestration cost-effectively. This goal is consistent with DOE objectives to provide economically competitive and environmentally safe options to offset projected growth in U.S. baseline emissions of greenhouse gases after 2010, achieve the long-term goal of$10/ton of avoided net costs for carbon sequestration, and provide half of the required reductions in global greenhouse gases by 2025. Other potential benefits of the demonstration include developing a passive technology for water treatment for trace metal and nutrient release reductions, using power plant by-products to improve coal mine land reclamation and carbon sequestration, developing wildlife habitat and green-space around production facilities, generating Total Maximum Daily Load (TMDL) credits for the use of process water, and producing wood products for use by the lumber and pulp and paper industry. Project activities conducted during the five year project period include: Assessing tree cultivation and other techniques used to sequester carbon; Project site assessment; Greenhouse studies to determine optimum plant species and by-product application; Designing, constructing, operating, monitoring, and evaluating the CCWESTRS system; and Reporting (ongoing). The ability of the system to sequester carbon will be the primary measure of effectiveness, measured by accessing survival and growth response of plants within the CCWESTRS. In addition, costs associated with design, construction, and monitoring will be evaluated and compared to projected benefits of other carbon sequestration technologies. The test plan involves the application of three levels each of two types of power plant by-products--three levels of FGD gypsum mulch, and three levels of ash pond irrigation water. This design produces nine treatment levels which are being tested with two species of hardwood trees (sweet gum and sycamore). The project is examining the effectiveness of applications of 0, 8-cm, and 15-cm thick gypsum mulch layers and 0, 13 cm, and 25 cm of coal fly ash water for irrigation. Each treatment combination is being replicated three times, resulting in a total of 54 treatment plots (3 FGD gypsum levels X 3 irrigation water levels x 2 tree species x 3 replicates). Survival and growth response of plant species in terms of sequestering carbon in plant material and soil will be the primary measure of effectiveness of each treatment. Additionally, the ability of the site soils and unsaturated zone subsurface materials will be evaluated for their effectiveness at treating the irrigation water for various pollutants

BCL11A enhancer edited hematopoietic stem cells persist in rhesus monkeys without toxicity

Gene editing of the erythroid-specific BCL11A enhancer in hematopoietic stem and progenitor cells (HSPCs) from sickle cell disease (SCD) patients induces fetal hemoglobin (HbF) without detectable toxicity as assessed by mouse xenotransplant. Here, we evaluated autologous engraftment and HbF induction potential of erythroid-specific BCL11A enhancer edited HSPCs in four non-human primates. We utilized a single guide RNA (sgRNA) with identical human and rhesus target sequences to disrupt a GATA1 binding site at the BCL11A +58 erythroid enhancer. Cas9 protein and sgRNA ribonucleoprotein complex (RNP) was electroporated into rhesus HSPCs, followed by autologous infusion after myeloablation. We found that gene edits persisted in peripheral blood (PB) and bone marrow (BM) for up to 101 weeks similarly for BCL11A enhancer or control locus (AAVS1) targeted cells. Biallelic BCL11A enhancer editing resulted in robust gamma-globin induction, with the highest levels observed during stress erythropoiesis. Indels were evenly distributed across PB and BM lineages. Off-target edits were not observed. Non-homologous end-joining repair alleles were enriched in engrafting HSCs. In summary, we find that edited HSCs can persist for at least 101 weeks post-transplant, and biallelic edited HSCs provide substantial HbF levels in PB red blood cells, together supporting further clinical translation of this approach

Dysregulation of Gene Expression in a Lysosomal Storage Disease Varies between Brain Regions Implicating Unexpected Mechanisms of Neuropathology

The characteristic neurological feature of many neurogenetic diseases is intellectual disability. Although specific neuropathological features have been described, the mechanisms by which specific gene defects lead to cognitive impairment remain obscure. To gain insight into abnormal functions occurring secondary to a single gene defect, whole transcriptome analysis was used to identify molecular and cellular pathways that are dysregulated in the brain in a mouse model of a lysosomal storage disorder (LSD) (mucopolysaccharidosis [MPS] VII). We assayed multiple anatomical regions separately, in a large cohort of normal and diseased mice, which greatly increased the number of significant changes that could be detected compared to past studies in LSD models. We found that patterns of aberrant gene expression and involvement of multiple molecular and cellular systems varied significantly between brain regions. A number of changes revealed unexpected system and process alterations, such as up-regulation of the immune system with few inflammatory changes (a significant difference from the closely related MPS IIIb model), down-regulation of major oligodendrocyte genes even though white matter changes are not a feature histopathologically, and a plethora of developmental gene changes. The involvement of multiple neural systems indicates that the mechanisms of neuropathology in this type of disease are much broader than previously appreciated. In addition, the variation in gene dysregulation between brain regions indicates that different neuropathologic mechanisms may predominate within different regions of a diseased brain caused by a single gene mutation

Compressed representation of a partially defined integer function over multiple arguments

In OLAP (OnLine Analitical Processing) data are analysed in an n-dimensional cube. The cube may be represented as a partially defined function over n arguments. Considering that often the function is not defined everywhere, we ask: is there a known way of representing the function or the points in which it is defined, in a more compact manner than the trivial one

The spotted gar genome illuminates vertebrate evolution and facilitates human-teleost comparisons

To connect human biology to fish biomedical models, we sequenced the genome of spotted gar (Lepisosteus oculatus), whose lineage diverged from teleosts before teleost genome duplication (TGD). The slowly evolving gar genome has conserved in content and size many entire chromosomes from bony vertebrate ancestors. Gar bridges teleosts to tetrapods by illuminating the evolution of immunity, mineralization and development (mediated, for example, by Hox, ParaHox and microRNA genes). Numerous conserved noncoding elements (CNEs; often cis regulatory) undetectable in direct human-teleost comparisons become apparent using gar: functional studies uncovered conserved roles for such cryptic CNEs, facilitating annotation of sequences identified in human genome-wide association studies. Transcriptomic analyses showed that the sums of expression domains and expression levels for duplicated teleost genes often approximate the patterns and levels of expression for gar genes, consistent with subfunctionalization. The gar genome provides a resource for understanding evolution after genome duplication, the origin of vertebrate genomes and the function of human regulatory sequences

Cell activation-based screening of natively paired human T cell receptor repertoires

Adoptive immune therapies based on the transfer of antigen-specific T cells have been used successfully to treat various cancers and viral infections, but improved techniques are needed to identify optimally protective human T cell receptors (TCRs). Here we present a high-throughput approach to the identification of natively paired human TCRα and TCRβ (TCRα:β) genes encoding heterodimeric TCRs that recognize specific peptide antigens bound to major histocompatibility complex molecules (pMHCs). We first captured and cloned TCRα:β genes from individual cells, ensuring fidelity using a suppression PCR. We then screened TCRα:β libraries expressed in an immortalized cell line using peptide-pulsed antigen-presenting cells and sequenced activated clones to identify the cognate TCRs. Our results validated an experimental pipeline that allows large-scale repertoire datasets to be annotated with functional specificity information, facilitating the discovery of therapeutically relevant TCRs