Immunohistochemical evidence for the reduced synaptic proteins at 6-months of age in the CA1 region of the hippocampus.

<p>Hippocampal brain sections from wild-type (WT) and APΔE9/RanBP9 triple transgenic mice were stained with antibodies against rab3a, gap43, drebrin and chromogranin. A qualitative difference is clearly seen with reduced immunoreactive puncta in the triple transgenic mice compared to WT brains for all the four synaptic proteins (red). The neuronal nuclei are stained blue. Scale bar: 100 µm.</p

RanBP9 overexpression does not alter synaptic protein levels in the cortex and hippocampus at 3- months of age.

<p><b>A:</b> Shows an immunoblotting analysis of rab3A, gap43, drebrin, chromogranin and the house keeping gene actin in brain samples from cortex and hippocampus. Brain homogenates from RanBP9 transgenic (Ran), APΔE9 double transgenic (Dbl), APΔE9/RanBP9 triple transgenic (Tpl) and age-matched wild-type (WT) control mice at 3-months of age were subjected to SDS-PAGE electrophoresis and probed with their respective antibodies. Flag specific monoclonal antibody detected flag-tagged exogenous RanBP9 in the RanBP9 single transgenic and APΔE9/RanBP99 triple transgenic mice only. Actin was used as loading control. The numbers on the left side indicate the molecular weights of each protein. <b>B:</b> Image J quantitation and normalization to actin levels showed no changes in the levels of any of the synaptic proteins at 3 months. The data are mean±SEM, n = 6 for WT and RanBP9 single transgenic, and n = 8 for APΔE9 and APΔE9/RanBP9 genotypes.</p

RanBP9 overexpression does not alter synaptic protein levels in the cortex and hippocampus at 4- months of age.

<p><b>A:</b> Shows an immunoblotting analysis of rab3A, gap43, drebrin, chromogranin and the house keeping gene actin in brain samples from cortex and hippocampus. Brain homogenates from RanBP9 transgenic (Ran), APΔE9 double transgenic (Dbl), APΔE9/RanBP9 triple transgenic (Tpl) and age-matched wild-type (WT) control mice at 4-months of age were subjected to SDS-PAGE electrophoresis and probed with their respective antibodies. Flag specific monoclonal antibody detected flag-tagged exogenous RanBP9 in the RanBP9 single transgenic and APΔE9/RanBP99 triple transgenic mice only. Actin was used as loading control. The numbers on the left side indicate the molecular weights of each protein. <b>B:</b> Image J quantitation and normalization to actin levels showed no changes in the levels of any of the synaptic proteins at 4 months. The data are mean±SEM, n = 6 for WT and RanBP9 single transgenic, and n = 8 for APΔE9 and APΔE9/RanBP9 genotypes.</p

Additional file 1: Table S1. of Effect of dexamethasone on hypothalamic expression of appetite-related genes in chickens under different diet and feeding conditions

The composition and nutrient levels of the experimental diets for chickens (air dry basis). (DOC 42 kb

<i>In silico</i> discovery of potential sodium–glucose cotransporter-2 inhibitors from natural products for treatment of heart failure via molecular docking and molecular dynamics simulation approach

Heart failure (HF) is the end stage of cardiovascular disease. Because of its complex condition and poor prognosis, HF has become an important public health problem in the world. Sodium–glucose cotransporter-2 (SGLT2) is a member of the glucose transporter family. Recently, SGLT2 inhibitors have been applied to treat HF. In this study, the main aim was to discover natural SGLT2 inhibitor from Chinese herbs through docking-based virtual screening. Totally 113 natural compounds of potential SGLT2 inhibitor were identified, which displayed docking affinity higher than six approved inhibitors (dapagliflozin (IC50 = 4.9 nM), canagliflozin (IC50 = 4.4 nM 6.7), ipragliflozin (IC50 = 7.4 nM), empagliflozin (IC50 = 3.1 nM), tofogliflozin (IC50 = 4 nM) and luseogliflozin (IC50 = 2.3 nM)) through docking-based virtual screening. Then, the top three hits (ZINC70455591, ZINC85594065 and ZINC14588133) and six known inhibitors were selected for molecular dynamics simulation and the binding free energy calculation using molecular mechanics Poisson–Boltzmann surface area to demonstrate the stability and affinity of docked complexes. These results pointed out that the three docked complexes were stabilized and the chosen compounds were tightly adhering to the binding site of SGLT2. Besides, pharmacokinetic properties of the selected compounds showed those natural compounds may be potential drug candidates. This study may be contributed to further in vitro and in vivo validation and the development of novel SGLT2 inhibitor for treating HF. Communicated by Ramaswamy H. Sarma</p

Data_Sheet_1_Determining the nutritional values of new corn varieties on pigs and broilers.docx

The objective of this study was to evaluate the nutritional values of three new corn varieties (high-iron corn, cadmium-resistant corn, low-phytate phosphorus corn) cultivated with molecular marker-assisted selection breeding technique fed to growing pigs and broilers. Exp. 1 was conducted to compare the nutritional values of high-iron corn, high-chromium corn, low-phytate phosphorus corn and conventional corn fed to growing pigs based on a 15 × 2 Youden square design. Exp.2 was conducted to compare the nutritional values of high-iron corn, low-phytate phosphorus corn and conventional corn fed to broilers based on a completely randomized design. Parameters including nutrient digestibility, available energy and amino acids, and mineral deposition were measured. The results shows that the iron content in the high-iron corn and the cadmium content in the cadmium-resistant corn were 29.608 mg/kg and 0.0057 mg/kg, respectively, both were greater than those in the other three kinds of corns. When fed to growing pigs, the neutral detergent fiber digestibility of the high-iron corn group was lower than that of the conventional corn group (p < 0.05), and the acid detergent fiber digestibility of the high-iron group and the low-phytate phosphorus corn group was lower than that of the conventional corn group (p < 0.01). In addition, the digestible energy value of the high-iron corn in growing pigs was lower than that of the conventional corn (p < 0.05). When fed to broilers, it was observed that the tibia length of the low-phytate phosphorus corn group and the high-iron corn group was lower than that of the conventional corn group (p < 0.05). Moreover, the iron emission in feces of broilers fed the low-phytate phosphorus corn was lower than those fed the conventional corn and the high-iron corn (p < 0.05). In conclusion, modern breeding techniques could provide new plant ingredients which have potential benefits to pig and broiler production, but the comprehensive effects may be better when applied to growing pigs considering growth performance and environment effects. The breeding techniques related to the current study rarely changed the available energy values of the corn in growing pigs and broilers.</p

Components of aggregating lysozyme.

<p>Left: AFM image showing various structures seen in aggregating lysozyme on mica substrate: (a) colloidal spheres, (b) primary fibers, (c) compound fibers, (d) amorphous aggregates, (e) a continuous layer of protein monomers bound to substrate. Right: After 1∶1000 dilution the continuous protein layer is no longer present. Colloidal spheres are still seen (a), as are numerous discrete particles (b) with a height of nm (n = 20), probably representing individual lysozyme monomers.</p

Transition to helical structure is inhibited by high salt concentration.

<p>Lysozyme aggregating in buffer with no salt (left) or 30 mM NaCl (center) demonstrate transformation of virtually all fibers to helical structure after incubation for 11 days. The helical curve of the lysozyme fiber bound to the substrate typically appears from above as a sinusoid (left, inset) although other patterns such as twisted ribbons are occasionally seen. The triangular symbols show the length of one turn of the helix is approximately.4m. Fibers forming in buffer with 150 mM NaCl (right) demonstrate virtually no helical structure even after 31 days.</p

Effect of time and salt concentration on fiber helicity.

<p>Newly formed fibers are linear at all salt concentrations; at zero and 30 mM NaCl the fraction of fibers displaying helical morphology increases consistently with time until all fibers are helical. This change occurs more rapidly at lower salt concentrations. At 150 nM NaCl essentially all fibers remain linear.</p

Gel formation in lysozyme.

<p>AFM images of lysozyme amyloid fiber network after 30 days incubation. The mixture is a firm gel at this point. (A) Lysozyme gel, undiluted, showing dense network structure. (B) When diluted with water 1∶100 and gently mixed, the gel swells and can be applied to the substrate in a monolayer, showing a range of fiber sizes, bound together at points of contact. (C) Detail of B, showing occasional merging of fibers at points of contact. (D) Gel was diluted in water (1∶10,000) and vortexed ×30 sec. Gel fibers are broken into short fragments but remain joined at points of contact. This indicates development of strong covalent linkages between fibers, typical of an irreversible macromolecular network or IRMAN <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094789#pone.0094789-Terech1" target="_blank">[8]</a>.</p