The amyloid precursor protein has a flexible transmembrane domain and binds cholesterol

C99 is the transmembrane carboxyl-terminal domain of the amyloid precursor protein that is cleaved by γ-secretase to release the amyloid-β polypeptides, which are associated with Alzheimer's disease. Nuclear magnetic resonance and electron paramagnetic resonance spectroscopy show that the extracellular amino terminus of C99 includes a surface-embedded "N-helix" followed by a short "N-loop" connecting to the transmembrane domain (TMD). The TMD is a flexibly curved α helix, making it well suited for processive cleavage by γ-secretase. Titration of C99 reveals a binding site for cholesterol, providing mechanistic insight into how cholesterol promotes amyloidogenesis. Membrane-buried GXXXG motifs (G, Gly; X, any amino acid), which have an established role in oligomerization, were also shown to play a key role in cholesterol binding. The structure and cholesterol binding properties of C99 may aid in the design of Alzheimer's therapeutics.     

Organic Fertilizer Source and Application Method Impact Ammonia Volatilization

ABSTRACT

Ammonia (NH₃) volatilization from fertilizer applications reduces efficiency and poses environmental hazards. This study used semi-open static chambers to measure NH₃ volatilization from organic fertilizers (feather meal, blood meal, fish emulsion, cyano-fertilizer) to evaluate the impacts of fertilizer source, application method, and rate on NH₃ volatilization. In 2014, two application rates (28 and 56 kg N ha^?1) were applied to lettuce (Lactuca sativa L.). Solid fertilizers (feather meal, blood meal) were preplant applied in a subsurface band, whereas liquid fertilizers (fish emulsion, cyano-fertilizer) were applied weekly through drip irrigation beginning two weeks after transplanting. In 2015, a single application rate (28 kg N ha^?1) was applied to cucumber (Cucumis sativus L.). Solid fertilizers were applied in either subsurface or surface bands. There was a significant difference in NH₃ volatilization among fertilizers, but there was little difference between application rates. Liquid fertilizers had lower NH₃ emissions than solid fertilizers due to their timing and placement. In 2014, blood meal at 56 kg N ha^?1 and feather meal at both rates had the highest NH₃ fluxes. In 2015, surface-banded blood and feather meal had the highest NH₃ fluxes. Fertilizer decisions for organic systems should consider NH₃ emission losses and practices for their reduction.     

Antioxidant Bioactive Compounds and Spoilage Microorganisms of Wax Apple (Syzygium samarangense) during Room Temperature Storage

Postharvest deterioration of wax apple leads to unacceptable appearances, physical and quality losses that raise serious concerns commercially. Bioactive compounds of wax apple and spoilage microorganisms were studied at room temperature storage (23 ± 1 °C). Antioxidant activity and total phenolic content increased up to a maximum of 1.56 mg AAE/100 g and 88.37 mg GAE/100 g, while prolonged storage resulted in 70.0% and 33.6% loss, respectively. Further, 80% loss in vitamin C content was observed from an initial value of 21.63 mg AAE/100 g during storage. Bacterial isolates of Enterobacter sakazakii, Klebsiella pneumoniae, Klebsiella planticola, Pantoea agglomerans, Chromobacterium violaceum, and Streptomyces roseochromogenus with microbial load of 1.19 * 10⁸ CFU/g fresh weight, and fungal isolates of Penicillium purpurogenum, Mucor hiemalis, Aspergillus niger, Aspergillus fumigatus, and Candida tropicalis with a total load of 5.64 * 10⁷ CFU/g fresh weight were identified as spoilage organisms. Bacterial isolates of Klebsiella pneumoniae was the most prevalent at 47.78%, occurring 21 times, while Penicillium purpurogenum was the most prevalent fungi, occurring 5 times at 38.46%. Results have significant implications on measures to preserve quality of wax apples during storage by adoption of advanced postharvest handling and processing approaches.     

Expression of CD3 and CD11b antigens on blood and mammary gland leukocytes and bacterial survival in milk of cows with experimentally induced Staphylococcus aureus mastitis.

OBJECTIVES: To differentiate early (1 to 8 days) from late (9 to 14 days) inflammatory phases and assess relationships between leukocyte phenotype and bacterial recovery in cows with Staphylococcus aureus-induced mastitis. ANIMALS: 10 first-lactation Holstein cows. PROCEDURE: Blood and milk samples were collected from 4 or 6 cows before and after intramammary infusion of sterile broth or S. aureus, respectively. Flow cytometric expression of CD3 and CD11b antigens on blood and milk leukocytes, leukocyte differential counts, bacterial counts in milk, and somatic cell counts were determined longitudinally. RESULTS: Density of CD3 molecules decreased on blood lymphocytes and increased on milk lymphocytes after infusion of bacteria. Density of CD11b molecules on lymphocytes and phagocytes and percentage of CD11b+ lymphocytes in milk increased significantly after infusion; maximum values were achieved during the early inflammatory phase. Density of CD3 and CD11b molecules on milk lymphocytes and macrophages, respectively, 1 day after inoculation were negatively correlated with bacterial recovery on day 1 and days 9 to 14, respectively. Density of CD11b molecules on milk macrophages and the ratios of phagocyte to lymphocyte percentages and polymorphonuclear cell to macrophage percentages in milk differentiated the early from the late inflammatory phase. CONCLUSIONS AND CLINICAL RELEVANCE: Activation of bovine mammary gland macrophages and T cells in response to intramammary infusion of S. aureus was associated with an inability to culture this bacterium from milk. Identification of specific inflammatory phases of S. aureus-induced mastitis in cows may allow for the design of more efficacious treatment and control programs.