Comparison of the amyloid burden in the Tg brains by the CG extracts.

<p>(A) The amyloid plaques were identified by immunohistochemistry (IH) and H&E counterstaining. The CG extract-treated (EA-CG fraction) and control Tg brain sections were formalin fixed and subjected to immunohistochemistry with the 6E10 antibody that interacts with the Aβ1–16 epitope (monoclonal antibody) in the hippocampus (a, control; b, EA-CG treated) and cortex (e, control; f, EA-CG-treated). Concomitant with the IH staining, other hippocampal (n = 6) (c, vehicle; d, EA-CG-treated) and cortical (n = 6) (g, vehicle; h, EA-CG-treated) sections from the Mo/Hu APPswe PS1dE9 mice were subjected to H&E counterstaining. The scale bar indicates 200 μm (a through h). The amyloid load (% of stained area) as a quantitation result was decreased by the CG extracts (B. hippocampus area; C. cortex area). The area in the hippocampus (D, vehicle alone vs. EA-CG) and cortex (E, vehicle alone vs. EA-CG) was quantified by comparing the morphometric analysis of the Thioflavin-S-stained area between the vehicle-treated vs. EA-CG-treated Mo/Hu APPswe PS1dE9 brain sections. The values (%) shown were the means ± S.E.M. *p < 0.05, Tg vs. Tg+EA-CG.</p

Comparison of Soluble Aβ42/40 by the CG extracts on Tg mice brain.

<p>The cerebral Aβ42(A) and Aβ40 (B) load was quantitated by capturing antigen such as Amyloid beta 42 and/or Amyloid beta 40 mediated Enzyme-Linked Immunosorbent Assay (ELISA) kit (Invitrogen, CA) using brain lysates (i.e., total brain lysate of hippocampus and neocortex area) from the EA-CG-treated or vehicle (PBS)-treated Tg (Mo/Hu APPswe PS1dE9) mice. The cerebral Aβ42 concentrations in the EA-CG-treated mice were significantly reduced compared to the vehicle-treated controls (**P < 0.01). The values shown were the means ± S.E.M. (n = 10 per group).</p

Additional file 1: of Rubus crataegifolius Bunge regulates adipogenesis through Akt and inhibits high-fat diet-induced obesity in rats

Effects of supplementing RCB on body weight gain, and serum profiles in rats fed a high fat diet for two weeks. (A) Weight gain, (B) serum triglyceride level, and (C) total cholesterol level. The values are expressed as the mean ± SD. The bars showing different letters indicate significant differences among each group of bars, according to Duncan’s test; *p < 0.05. (ZIP 90 kb

Effect of CG extracts on liver toxicity using aspartate/alanine aminotransferase activity in Tg mice.

<p>The Mo/Hu APPswe PS1dE9 mice were fed with or without EA-CG (50 mg/kg BW) for 6 months. The brain were isolated from the EA-CG treated and control (PBS) groups (n = 3 mice per each group). After the brain was divided in half, we tested the gene expression profile of the inflammatory cytokines IL-1α (A) and IFN-γ (B) as markers of the cellular response by semi-quantitative RT-PCR. The fold change was presented after normalization to the housekeeping gene β-actin). *p < 0.05; n.s. mean not significant. The levels of Aspartate aminotransferase activity (AST, Units/L) and Alanine aminotransferase activity (ALT, Units/L)) were measured in the sera by centrifugation after obtain mice blood from each group, and then the relative value (U/L) of ALT (C) or AST (D) as hepatotoxicity markers were measured and presented after comparison to the controls (PBS groups), according to the manufacturer’s instructions. The values were presented as the means ± S.E.M (n = 3 mice per each group). n.s. mean not significant.</p

Assessment workflow of novel Anti-AD effectiveness using Maysin and its flavonoid derivative compounds on APP/PS1 AD mice model.

<p>Complication of traumatic brain injury is associated with inflammation and neural cell death from induction of Aβ 42/40 formation likely due to unfolded cerebral plaque or fibril form which results from genetic alteration in APPswe and PSEN1dE9 in the 85Dbo/J mice model for Experimental AD study. Experimental design for treatment and sampling procedure as the objectives in animal model to understand anti-AD of Maysin flavonoid with respect to prevention and/or prophylactic effect against Aβ40/42 mediated neuronal-toxicity and functional delineation as the assessment workflow in animal study are presented.</p

Effect of Centipedegrass (CG) extract on humoral immune response in APP/PS1 transgenic (Tg) mice.

<p>The CG ethylacetate (EA-CG) fraction contains five flavoglycosides, including Maysin and other phytochemicals, as described in a previous study [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169509#pone.0169509.ref020" target="_blank">20</a>]. Mouse sera were isolated from the EA-CG-treated and control Tg animal subjects (Mo/Hu APPswe PS1dE9) after feeding with admixture in diet food for 6 months (n = 10 mice per each group; all male and 10 weeks old mice). (A, B) To measure the beta amyloid dependent antibody response, using a sandwich ELISA, quantitation was performed utilize the Aβ1–42 synthetic peptide as the coating antigen and HRP-conjugated secondary IgG or IgM as a secondary antibody. Immunoglobulin IgG (A) and Immunoglobulin IgM (B), markers of an immune response against the EA-CG extract, were measured by ELISA, which was described in detail in the Materials and Methods. The values were presented as the means ± S.E.M. *p < 0.05 means statistically significant.</p

Antioxidant capacities, total phenolic and flavonoids content of the CCC extracts.

a<p>DPPH, DPPH radical scavenging activity;</p>b<p>HRSA, hydroxyl radial scavenging activity;</p>c<p>SRSA, superoxide anion radical scavenging activity;</p>d<p>TPC, total phenolic acid. Total phenolic acid and total flavonoid content are expressed as milligrams of quercetin equivalent (QE)/g of extract.</p>x–z<p>The values are presented as the mean ± SD. P<0.01 represents a significant difference between the samples (n = 4).</p><p>Antioxidant capacities, total phenolic and flavonoids content of the CCC extracts.</p

Effects of CCC on the regulation of Akt and GSK3β during adipocyte differentiation.

<p>Confluent 3T3-L preadipocytes were treated either with vehicle or CCC (0, 40, or 150 µg/ml) in the differentiation medium during day 0 to 7 of adipogenesis. (A) The protein level and the degree of Akt phosphorylation were analyzed in 3T3-L1 cells on days 3, 5, and 7 after the induction of differentiation. These experiments were conducted as independent experiments in triplicate. The data represent the mean ± SD. *p<0.05. **P<0.01. (B) Effect of CCC on GSK3β activation in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with CCC extracts at the indicated concentrations, and the phosphorylation levels of GSK3β were determined by western blot analysis. The data are presented as the mean ± SD values of at least three independent experiments. *p<0.05. **P<0.01. (C) Effect of CCC on Akt phosphorylation in A549 lung cancer cells. A549 lung cancer cells were treated with 40 or 150 µg/ml CCC in the absence or presence of insulin (50 ng/ml) for 12 h. The representative Western bolts from one of three independent experiments were shown. (D) Effects of the PI3K/Akt inhibitor LY294002 on the CCC-induced inhibition of adipocyte differentiation in 3T3-L1 cells. 3T3-L1 cells were treated with CCC during differentiation in the presence or absence of 10 µM LY294002. The intracellular lipid accumulation was measured using a triglyceride assay. The data are expressed as the mean ± SD of three independent experiments. *P<0.05. (E) Effect of CCC on insulin-stimulated glucose uptake. Glucose uptake activity was analyzed by measuring of 2-NBDG in differentiated 3T3-L1 cells. 3T3-L1 cells were induced to differentiate into adipocytes for 6 days in DMI medium without or with CCC (10, 40, and 150 µg/m), and glucose uptake was then measured. The data are represented as the value relative to that of the undifferentiated cells. The data are presented as the mean ± SD from 3 independent experiments. *p<0.05. **P<0.01.</p

Effects of CCC extracts on the body weight of HFD-induced obese rats.

<p>The results are presented as the mean ± SD. RD, regular diet-fed rat; HFD, high-fat diet-fed rat; HFD+CCC (60 mg/kg), high-fat diet containing 60 mg/kg BW CCC-fed rat; HFD+CCC (200 mg/kg), high-fat diet containing 200 mg/kg BW CCC-fed rat. The body weight was measured twice a week. Results are presented as the means ± SD.</p>a<p>P<0.05,</p>b<p>P<0.01 compared to the HFD group;</p><p>**P<0.01 compared to the RD group.</p><p>Effects of CCC extracts on the body weight of HFD-induced obese rats.</p

Treatment with CCC decreased the adipose tissue of the HFD-induced obese rats.

<p>The epididymal fat and perirenal fat weights were significantly decreased in the HDF+CCC (200 mg/kg) group compared to the HFD group. (A) Effects of CCC on epididymal fat weight. The weights of the epididymal fatty tissue were calculated by dividing the fatty tissue weight by the body weight (fatty tissue/body weight x 100). The values are expressed as the mean ± SD. The bars with different letters are significantly different (*p<0.05). (B) Effects of CCC on perirenal fat weight. The values are expressed as the mean ± SD. The bars showing different letters indicate significant differences among each group of bars, according to Duncan's test; *P<0.05. The means sharing a common letter do not significantly differ. (C) Morphology of the fat tissue altered by CCC in the HFD-induced obese rats. (D) Histological staining of epididymal adipose tissue. Epididymal adipose tissues were isolated and stained with hematoxylin and eosin (H&E), then examined microscopically. Representative micrographs showing the reduced adipocyte size in the HFD+CCC (200 mg/kg) group are shown. (E) Representative H&E-stained liver section. Liver tissues were isolated and stained with H&E, then examined microscopically. Representative micrographs showing the reduced number and size of lipid droplets in the HFD+CCC (200 mg/kg) group are shown.</p