Biallelic loss of LDB3 leads to a lethal pediatric dilated cardiomyopathy.

Autosomal dominant variants in LDB3 (also known as ZASP), encoding the PDZ-LIM domain-binding factor, have been linked to a late onset phenotype of cardiomyopathy and myofibrillar myopathy in humans. However, despite knockout mice displaying a much more severe phenotype with premature death, bi-allelic variants in LDB3 have not yet been reported. Here we identify biallelic loss-of-function variants in five unrelated cardiomyopathy families by next-generation sequencing. In the first family, we identified compound heterozygous LOF variants in LDB3 in a fetus with bilateral talipes and mild left cardiac ventricular enlargement. Ultra-structural examination revealed highly irregular Z-disc formation, and RNA analysis demonstrated little/no expression of LDB3 protein with a functional C-terminal LIM domain in muscle tissue from the affected fetus. In a second family, a homozygous LDB3 nonsense variant was identified in a young girl with severe early-onset dilated cardiomyopathy with left ventricular non-compaction; the same homozygous nonsense variant was identified in a third unrelated female infant with dilated cardiomyopathy. We further identified homozygous LDB3 frameshift variants in two unrelated probands diagnosed with cardiomegaly and severely reduced left ventricular ejection fraction. Our findings demonstrate that recessive LDB3 variants can lead to an early-onset severe human phenotype of cardiomyopathy and myopathy, reminiscent of the knockout mouse phenotype, and supporting a loss of function mechanism

Phosphorus Dynamics in Broiler Litter-Amended Soils

Because land application of broiler litter is commonly made in the spring before cropping season and in the fall, it is essential to understand how environmental variations, such as temperature, affect the phosphorus (P) dynamics. A laboratory incubation study was conducted at Waste Management and Forage Research Unit, USDAARS with the objective of determining the effects of temperature and soil properties on water-soluble P (WSP), Mehlich 3 P, and P fractions in broiler litter-amended soils. Broiler litter was mixed with three soils (Leeper silty clay, Grenada silt loam, and Ruston sandy loam) at the rate of 10 Mg ha-1. The mixtures were incubated at 18 °C, 25 °C, and 32 °C for up to 90 days and repeated three times. Soil samples were taken at eight time intervals (2, 5, 10, 15, 30, 60, and 90 days) and analyzed for water-soluble P and Mehlich 3-extractable P (MEP). Phosphorus also was fractionated chemically into inorganic and organic components by sequential extraction for soil samples taken at the end of incubation period (90 days). Water-soluble P decreased rapidly after a 15-day incubation in all soils for all temperatures and followed to a steady state to the end of incubation period. Averaged across temperatures, broiler litter application resulted in a significant increase in all fractions compared with controls. However, the increase was greater for inorganic than for organic fractions. The concentration of calcium (Ca)-P fraction was highest in calcareous Grenada soil, which suggests that the presence of carbonates influence the fate of P from applied broiler litter. The coarse textured Ruston contained a greater concentration of water-soluble P (WSP) than the other soils. At 32 °C, the concentration of plantavailable P fractions [bicarbonate inorganic P (IP) and hydroxide IP] significantly increased and WSP decreased. It is likely that the higher temperature (32 °C) promotes higher biological activity and lower water-soluble P than lower temperature

Effect of turning frequency and season on composting materials from swine high-rise facilities

Composting swine slurries has several advantages, liquid slurries are converted to solids at lower moisture, the total volume and weight of material is reduced and the stabilized product is more easily transported off-site. Despite this, swine waste is generally stored, treated and applied in its liquid form. High-rise finishing facilities (HRFF) permit liquid slurries to be converted to solids which are partially decomposed underneath the HRFF and then finished in compost windrows. The purpose of this study was to evaluate the effect of turning frequency and ambient weather conditions on biological, physical and chemical properties of composted slurry-woodchip mixtures from HRFF. Compost trials were conducted in either fall (FT) or spring (ST) and piles were turned once or three times per week or upon compost temperature reaching 65 °C. Physical, chemical and microbiological characteristics were measured over the course of 112 (FT) or 143 (ST) days of composting. Total carbon, total nitrogen (N) and inorganic N decreased in all piles. Ammonium decreased while nitrate increased in all piles (including unturned), but total N losses were greatest in piles turned more frequently during the ST. Microbial populations of nitrifiers were dominated by ammonia-oxidizing archaea (3.0 X 10^3–4.2 X 10^6 cells g^-1 compost) but ammonia oxidizing bacteria (below detection to 6.0 X cells g^-1 compost) varied in response to turning and compost temperature; denitrifiers were present in high concentrations throughout the process. Swine HRFF materials composted well in windrows regardless of turning frequency and despite significant differences in starting materials and low initial C/N. Volume reduction, low moisture and low readily degradable organic matter suggest that the finished compost would have lower transportation costs and should provide value as a soil conditioner

Nationally Coordinated Evaluation of Soil Nitrogen Mineralization Rate using a Standardized Aerobic Incubation Protocol

Aerobic incubation methods have been widely used to assess soil nitrogen (N) mineralization, but standardized protocols are lacking. A single silt loam soil (Catlin silt loam; fine-silty, mixed, superactive, mesic, Oxyaquic Arguidoll) was subjected to aerobic incubation at six USDA-ARS locations using a standardized protocol. Incubations were conducted at multiple temperatures, which were combined based on degree days (DD). Soil water was maintained at 60% waterfilled pore space (WFPS; constant) or allowed to fluctuate between 60 and 30% WFPS (cycle). Soil subsamples were removed periodically and extracted in 2 M potassium chloride (KCl); nitrate (NO3) and ammonium (NH4) concentrations in extracts were determined colorimetrically. For each location, the rate of soil organic-matter N (SOMN) mineralization was estimated by regressing soil inorganic N (Ni) concentration on DD, using a linear (zero-order) model. When all data were included, the mineralization rate from four datasets was not statistically different, with a rate equivalent to 0.5 mg N kg-1 soil day-1. Soil incubated at two locations exhibited significantly higher SOMN mineralization rates. To assess whether this may have been due to pre-incubation conditions, time-zero data were excluded and regression analysis was conducted again. Using this data subset, SOMN mineralization from five (of six) datasets was not significantly different. Fluctuating soil water reduced N-mineralization rate at two (of four) locations by an average of 50%; fluctuating soil water content also substantially increased variability. This composite dataset demonstrates that standardization of aerobic incubation methodology is possible

Protocols for Nationally Coordinated Laboratory and Field Research on Manure Nitrogen Mineralization

The National Program structure of USDA-ARS provides an opportunity to coordinate research on problems of national and global significance. A team of USDA-ARS scientists is conducting nationally coordinated research to develop predictions of manure N availability to protect water quality and improve farm solvency. Experimental design and research protocols were developed and used in common across all participating locations. Laboratory incubations are conducted at each location with a minimum of three soils, three temperatures, two wetting/drying regimes, and two manure treatments. A soil from the central United States (Catlin silt loam, fine-silty, mixed, superactive, mesic Oxyaquic Argiudoll) is used as an internal reference across all locations. Incubation data are compiled across locations to develop generalized predictions of manure nitrogen mineralization (Nmin). Field validation data are then obtained by monitoring nitrogen (N) transformations in manure-amended soil cores equipped with anion exchange resin to capture leached nitrate. This field data will be used to compare laboratory-based predictions with field observations of Nmin in each soil, climatic zone, and manure type represented. A Decision Support System will then be developed for predicting manure N mineralization across ranges in soil, climate, and manure composition. Protocols used by this research team are provided to 1) document the procedures used and 2) offer others detailed information for conducting research on nutrient transformation processes involving collaboration across locations or complementary research between laboratory and field environments