Simultaneous extraction and separation of oil, proteins, and glucosinolates from Moringa oleifera seeds.

Moringa oleifera is a worldwide cultivated edible and medicinal plant. Its seeds are rich in oil, proteins, and glucosinolates. A practical method was developed to simultaneously extract and separate the three groups of substances from M. oleifera seeds. Smashed seed material was loaded into columns with petroleum ether: ethanol 8:2 (PE-ethanol) and eluted sequentially with 4.8-fold PE-ethanol to extract oil, and 10.8-fold water to extract proteins and glucosinolates. More than 95% of oil, proteins, and glucosinolates were extracted. The extracts were separated automatically into ether (oil) phase and ethanol aqueous phase. The latter was further separated into proteins and glucosinolates by 70% ethanol precipitation. The main glucosinolate was identified by LC-MS as GLC (4-α-rhamnopyranosyloxy-benzyl glucosinolate). After purification, 22.3 g refined oil, 33.0 g proteins, and 5.5 g purified GLC from 100 g M. oleifera seeds were obtained. This study provides a simple and high-efficient method to utilize M. oleifera seeds.

Overexpressing STAMP2 attenuates adipose tissue angiogenesis and insulin resistance in diabetic ApoE-/- /LDLR-/- mouse via a PPARγ/CD36 pathway.

The aim of this study was to investigate whether overexpression of STAMP2 improves insulin resistance by regulating angiogenesis in adipose tissues. The characteristics of diabetic mice were measured by serial metabolite and pathology tests. Samples were obtained from epididymal, subcutaneous and brown adipose tissues. Histological and morphological analysis demonstrated that STAMP2 gene overexpression reduced adipocyte size, angiogenesis in epididymal and brown adipose tissues. On aortic ring assay, microvessels sprouting from aortas were significantly inhibited after STAMP2 gene overexpression. The cellular effect of STAMP2 on angiogenesis was explored in human umbilical vein endothelial cells (HUVECs) model. Correlation of STAMP2 and angiogenesis was validated by Ad-STAMP2 transfection and STAMP2 siRNA inhibition. In vitro, overexpression of STAMP2 significantly inhibited endothelial cell migration, tube formation. The effects of Ad-STAMP2 transfection on HUVECs were abolished by treatment with PPARγ antagonist GW9662 (2.5 µM), and the roles of STAMP2 siRNA on HUVECs were also reversed by treatment with PPARγ agonist rosiglitazone (RSG) (0.1 mM). RT-PCR indicated that STAMP2 could regulate levels of adhesion molecules, vascular endothelial growth factor A and CD36. The expression of PPARγ and CD36 was decreased when STAMP2 was inhibited by siRNA, while PPARγ and CD36 were highly expressed after overexpression of STAMP2. Our results suggested that STAMP2 gene overexpression may improve insulin resistance via attenuating angiogenesis in epididymal and brown adipose tissues through the PPARγ/CD36 signalling pathway.

Adipose-derived stem cells were impaired in restricting CD4+T cell proliferation and polarization in type 2 diabetic ApoE-/- mouse.

BACKGROUND: Atherosclerosis (AS) is the most common and serious complication of type 2 diabetes mellitus (T2DM) and is accelerated via chronic systemic inflammation rather than hyperglycemia. Adipose tissue is the major source of systemic inflammation in abnormal metabolic state. Pro-inflammatory CD4+T cells play pivotal role in promoting adipose inflammation. Adipose-derived stem cells (ADSCs) for fat regeneration have potent ability of immunosuppression and restricting CD4+T cells as well. Whether T2DM ADSCs are impaired in antagonizing CD4+T cell proliferation and polarization remains unclear. METHODS: We constructed type 2 diabetic ApoE-/- mouse models and tested infiltration and subgroups of CD4+T cell in stromal-vascular fraction (SVF) in vivo. Normal/T2DM ADSCs and normal splenocytes with or without CD4 sorting were separated and co-cultured at different scales ex vivo. Immune phenotypes of pro- and anti-inflammation of ADSCs were also investigated. Flow cytometry (FCM) and ELISA were applied in the experiments above. RESULTS: CD4+T cells performed a more pro-inflammatory phenotype in adipose tissue in T2DM ApoE-/- mice in vivo. Restriction to CD4+T cell proliferation and polarization was manifested obviously weakened after co-cultured with T2DM ADSCs ex vivo. No obvious distinctions were found in morphology and growth type of both ADSCs. However, T2DM ADSCs acquired a pro-inflammatory immune phenotype, with secreting less PGE2 and expressing higher MHC-II and co-stimulatory molecules (CD40, CD80). Normal ADSCs could also obtain the phenotypic change after cultured with T2DM SVF supernatant. CONCLUSION: CD4+T cell infiltration and pro-inflammatory polarization exist in adipose tissue in type 2 diabetic ApoE-/- mice. T2DM ADSCs had impaired function in restricting CD4+T lymphocyte proliferation and pro-inflammatory polarization due to immune phenotypic changes.

Mucin2 is Required for Probiotic Agents-Mediated Blocking Effects on Meningitic E. coli-Induced Pathogenicities.

Mucin2 (MUC2), an important regulatory factor in the immune system, plays an important role in the host defense system against bacterial translocation. Probiotics known to regulate MUC2 gene expression have been widely studied, but the interactions among probiotic, pathogens, and mucin gene are still not fully understood. The aim of this study was to investigate the role of MUC2 in blocking effects of probiotics on meningitic E. coli-induced pathogenicities. In this study, live combined probiotic tablets containing living Bifidobacterium, Lactobacillus bulgaricus, and Streptococcus thermophilus were used. MUC2 expression was knocked down in Caco-2 cells by RNA interference. 5-Aza-2'-deoxycytidine (5-Aza-CdR), which enhances mucin-promoted probiotic effects through inducing production of Sadenosyl- L-methionine (SAMe), was used to up-regulate MUC2 expression in Caco-2 cells. The adhesion to and invasion of meningitic E. coli were detected by competition assays. Our studies showed that probiotic agents could block E. coli-caused intestinal colonization, bacteremia, and meningitis in a neonatal sepsis and meningitis rat model. MUC2 gene expression in the neonatal rats given probiotic agents was obviously higher than that of the infected and uninfected control groups without probiotic treatment. The prohibitive effects of probiotic agents on MUC2-knockdown Caco-2 cells infected with E44 were significantly reduced compared with nontransfected Caco-2 cells. Moreover, the results also showed that 5- Aza-CdR, a drug enhancing the production of SAMe that is a protective agent of probiotics, was able to significantly suppress adhesion and invasion of E44 to Caco-2 cells by upregulation of MUC2 expression. Taken together, our data suggest that probiotic agents can efficiently block meningitic E. coli-induced pathogenicities in a manner dependent on MUC2.

Photocatalytic activities of Mo-doped Bi2WO6 three-dimensional hierarchical microspheres.

The Mo-doped Bi(2)WO(6) three-dimensional (3D) hierarchical microspheres from nanoplates have been synthesized by a hydrothermal route. The products were characterized in detail by multiform techniques: X-ray diffraction (XRD), energy-dispersive X-ray analysis (EDS), scanning electron microscopy (SEM), and UV-vis absorption spectrum. The results of the photocatalytic degradation of Rhodamine-B (RhB) in aqueous solution showed that molybdenum ions doping greatly improved the photocatalytic efficiency of Bi(2)WO(6) 3D hierarchical microspheres. The Mo-doped Bi(2)WO(6) microspheres with atomic ratio of Mo-W of 0.05 had the best activity in photodegradation of RhB in aqueous solution under 500 W Xe lamp light irradiation.

Photocatalytic activities of Cd-doped ZnWO4 nanorods prepared by a hydrothermal process.

The ZnWO(4) nanorods doped with cadmium ions have been successfully synthesized by a hydrothermal crystallization process. The products were characterized in detail by multiform techniques: X-ray diffraction (XRD), energy dispersive X-ray analysis (EDS), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The results of the photocatalytic degradation of rhodamine B (RhB) in aqueous solution showed that cadmium ions doping greatly improved the photocatalytic efficiency of ZnWO(4) nanorods. The Cd-doped ZnWO(4) nanorods with atomic ratio of Cd to Zn being 0.06 had the best activity in photo-degradation of RhB in aqueous solution under UV light irradiation, when the nanorods have prepared at pH 8.