Protein enrichment and digestion improvement of napiergrass and pangolagrass with solid-state fermentation.

BACKGROUND AND PURPOSE: Napiergrass (Pennisetum purpureum Schumacher) and pangolagrass (Digitaria decumbens Stent) are two major forage grasses for cow feeding. They possess high yields and high regeneration properties. Inoculation of cellulolytic microbes on herbage could enhance the protein content of herbage and promote digestibility in chickens. METHODS: Cellulolytic microbes were isolated from various sources and cultivated on napiergrass and pangolagrass with solid-state fermentation for protein enrichment and in vitro digestion improvement.The fermented napiergrass and pangolagrass were used as the main protein source in chicken diets to assess the feasibility for non-ruminants feed. RESULTS: After a 42-day fermentation period, napiergrass showed higher protein contents (13.4-13.9%) than those of pangolagrass(11.1-11.7%). The in vitro digestibility of pangolagrass increased from 5.29% to 20.4%, whereas that of napiergrass increased from 5.29% to 19.0%. The average feed conversion efficiencies of chickens were close to the traditional fodder using corn as the main ingredient. CONCLUSION: Inoculation of appropriate cellulolytic microbes to enrich protein content and improve in vitro digestibility of herbage with solid-state fermentation for chicken feed is the prospective technique for agriculture, animal husbandry, and substantial management.

Giant right atrial myxoma with pulmonary trunk dislodgement causing intermittent tricuspid obliteration and clinical manifestations of right heart failure.

The symptoms of myxomas depend on the size, mobility, and location of the tumor. A huge myxoma obstructing the tricuspid orifice can produce symptoms of tricuspid stenosis. In this case, a giant right atrial myxoma with intermittent tricuspid obliteration, presenting with clinical manifestations of right heart failure, is described. Three-dimensional reconstruction clearly identified the occlusive extent of the tricuspid orifice.

Microbial communities and bacterial diversity of spruce, hemlock and grassland soils of Tatachia Forest, Taiwan.

To evaluate the bacterial diversity of Tatachia Forest soils, 16S rDNA clone libraries of the spruce, hemlock and grassland soils were constructed. Further, the influence of physicochemical and biological properties of soil on microbial ecology, pH, moisture content, microbial population and biomass were also analyzed. The soil pH increased with the increasing of soil depth; whereas the microbial population, biomass, moisture content, total organic carbon and total nitrogen were reverse. Microbial populations were the highest in the summer season which also correlated with the highest moisture content. The phylogenetic analyses revealed that the clones from nine 16S rDNA clone libraries represented Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, Firmicutes, Gemmatimonadetes, Planctomycetes, Verrucomicrobia, candidate division TG1 and candidate division TM7. Members of Proteobacteria, Acidobacteria and Actinobacteria constituted 42.2, 35.1 and 7.8 % of the clone libraries, respectively; whereas the remaining bacterial divisions each comprised <3 %. The spruce site had the highest bacterial diversity among the tested sites, followed by the hemlock sites and the grassland sites with the least. The bacterial community is the more diverse in the organic layer than that in deeper horizons. Further, bacterial diversity through the gradient horizons was different, indicating that the bacterial diversity in the deeper horizons is not simply the diluted analogs of the surface soils and some microbes dominate only in the deeper horizons.

Thermo-tolerant phosphate-solubilizing microbes for multi-functional biofertilizer preparation.

In order to prepare the multi-functional biofertilizer, thermo-tolerant phosphate-solubilizing microbes including bacteria, actinomycetes, and fungi were isolated from different compost plants and biofertilizers. Except Streptomycesthermophilus J57 which lacked pectinase, all isolates possessed amylase, CMCase, chitinase, pectinase, protease, lipase, and nitrogenase activities. All isolates could solubilize calcium phosphate and Israel rock phosphate; various isolates could solubilize aluminum phosphate, iron phosphate, and hydroxyapatite. During composting, biofertilizers inoculated with the tested microbes had a significantly higher temperature, ash content, pH, total nitrogen, soluble phosphorus content, and germination rate than non-inoculated biofertilizer; total organic carbon and carbon-to-nitrogen ratio showed the opposite pattern. Adding these microbes can shorten the period of maturity, improve the quality, increase the soluble phosphorus content, and enhance the populations of phosphate-solubilizing and proteolytic microbes in biofertilizers. Therefore, inoculating thermo-tolerant phosphate-solubilizing microbes into agricultural and animal wastes represents a practical strategy for preparing multi-functional biofertilizer.

Methane and carbon dioxide emissions from closed landfill in Taiwan.

The atmospheric concentrations and emission rates of CH(4) and CO(2) were studied at three sites of the Fu-Der-Kan closed landfill and after as the multi-use recreational park in northern Taiwan. Atmospheric CH(4) and CO(2) concentrations of closed landfill were 1.7-4.6 and 324-409ppm, respectively. CH(4) and CO(2) emission rates ranged from 8.8 to 163mg m(-2)h(-1) and from 495 to 1531mg m(-2)h(-1), respectively. Diurnal variation was noted with higher values at night than those in daytime. After creation of the park, atmospheric CH(4) and CO(2) concentrations were 1.8-3.1 and 332-441ppm, respectively. CH(4) and CO(2) emission rates ranged from -1.1 to 2.3mg m(-2)h(-1) and from -135 to 301mg m(-2)h(-1), respectively. There were no notable diurnal variations in either atmospheric concentrations or emission rates.

Fraxetin inhibits the induction of anti-Fas IgM, tumor necrosis factor-alpha and interleukin-1beta-mediated apoptosis by Fas pathway inhibition in human osteoblastic cell line MG-63.

The survival of osteoblast cells is one of the determinants of the development of osteoporosis in patients with inflamed synovium, such as in rheumatoid arthritis (RA). By means of alkaline phosphatase (ALP) activity and osteocalcin ELISA assay, we have shown that fraxetin exhibits a significant induction of differentiation in the human osteoblast-like cell line MG-63. In addition, we also assessed whether fraxetin affects inflammatory cytokine-mediated apoptosis in osteoblast cells. TNF-alpha or IL-1beta enhance apoptotic DNA fragmentation in anti-Fas IgM-treated MG-63 cells by increasing Fas receptor expression. However, TNF-alpha or IL-1beta treatment alone does not induce apoptosis. Treatment of MG-63 cells with fraxetin not only inhibited anti-Fas IgM-induced apoptosis, but also blocked the synergetic effect of anti-Fas IgM with TNF-alpha or IL-1beta on cell death. The apoptotic inhibition of fraxetin is associated with inhibition of TNF-alpha and IL-1beta-mediated Fas expression and enhancement of FLIP expression, resulting in a decrease of caspase-8 and caspase-3 activation. These results indicate a potential use of fraxetin in preventing osteoporosis by inhibiting inflammatory cytokine-mediated apoptosis in osteoblast cells.

Bone morphogenetic protein-2 and -4 (BMP-2 and -4) mediates fraxetin-induced maturation and differentiation in human osteoblast-like cell lines.

Fraxetin (7,8-dihydroxy-6-methoxy coumarin), a coumarin derivative, was investigated for its effects on differentiation of osteoblasts. By means of alkaline phosphatase (ALP) activity and osteocalcin ELISA assay, we have shown that fraxetin exhibits a significant induction of differentiation in two human osteoblast-like cell lines, MG-63 and hFOB. Alkaline phosphatase and osteocalcin are phenotypic markers for early-stage differentiated osteoblasts and terminally differentiated osteoblasts, respectively. Our results indicated that fraxetin stimulated osteoblast differentiation at various stages (from osteoprogenitors to terminally differentiated osteoblasts). Induction of differentiation by fraxetin was associated with increased bone morphogenetic protein-2 (BMP-2) and BMP-4 productions. Addition of purified BMP-2 and BMP-4 proteins did not increase the upregulation of ALP activity and osteocalcin secretion by fraxetin, whereas the BMPs antagonist noggin blocked both fraxetin and BMP-2 and BMP-4 mediated ALP activity and osteocalcin secretion enhancement, indicating that BMP-2 and BMP-4 productions are required in fraxetin-mediated osteoblast maturation and differentiation. These findings are novel and may be important in the treatment and prevention of osteoporosis.

The anti-proliferative inhibition of ellipticine in human breast mda-mb-231 cancer cells is through cell cycle arrest and apoptosis induction.

Ellipticine, a cytotoxic plant alkaloid, is known to inhibit topoisomerase II. Here we report the mechanism of apoptosis induction and cell cycle arrest by ellipticine in human breast MDA-MB-231 cancer cells. Ellipticine treatment arrested MDA-MB-231 cells at the G2/M phase after 6 h of treatment. This effect was strongly associated with a concomitant decrease in the level of cyclin B1, Cdc25 and Cdc2, and increase in phospho-Cdc2 (Tyr15). In addition, ellipticine also induced apoptosis in MDA-MB-231 cells, as determined by using both DNA fragmentation and Annexin-V staining assay. Ellipticine increased the expression of Bax, but decreased the level of Bcl-2, Bcl-XL and X-linked inhibitor of apoptosis protein (XIAP), and subsequently triggered the mitochondrial apoptotic pathway (release of cytochrome c, and activation of caspase-9 and -3). In addition, pre-treatment of cells with caspase-9 inhibitor inhibited ellipticine-induced cell proliferation and apoptosis, indicating that caspase-9 activation was involved in MDA-MB-231 cell apoptosis induced by ellipticine. Taken together, our study suggests that the inhibition of cell cycle progression signaling and initiation of the mitochondrial apoptotic system may participate in the anti-proliferative activity of ellipticine in MDA-MB-231 cells.

Osthole-mediated cell differentiation through bone morphogenetic protein-2/p38 and extracellular signal-regulated kinase 1/2 pathway in human osteoblast cells.

The survival of osteoblast cells is one of the determinants of the development of osteoporosis in patients. Osthole (7-methoxy-8-isopentenoxycoumarin) is a coumarin derivative present in many medicinal plants. By means of alkaline phosphatase (ALP) activity, osteocalcin, osteopontin, and type I collagen, enzyme-linked immunosorbent assay, we have shown that osthole exhibits a significant induction of differentiation in two human osteoblast-like cell lines, MG-63 and hFOB. Induction of differentiation by osthole was associated with increased bone morphogenetic protein (BMP)-2 production and the activations of SMAD1/5/8 and p38 and extracellular signal-regulated kinase (ERK) 1/2 kinases. Addition of purified BMP-2 protein did not increase the up-regulation of ALP activity and osteocalcin by osthole, whereas the BMP-2 antagonist noggin blocked both osthole and BMP-2-mediated ALP activity enhancement, indicating that BMP-2 production is required in osthole-mediated osteoblast maturation. Pretreatment of osteoblast cells with noggin abrogated p38 activation but only partially decreased ERK1/2 activation, suggesting that BMP-2 signaling is required in p38 activation and is partially involved in ERK1/2 activation in osthole-treated osteoblast cells. Cotreatment of p38 inhibitor SB203580 [4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole] or p38 small interfering RNA (siRNA) expression inhibited osthole-mediated activation of ALP but only slightly affected osteocalcin production. In contrast, the production of osteocalcin induced by osthole was inhibited by the mitogen-activated protein kinase kinase inhibitor PD98059 (2'-amino-3'-methoxyflavone) or by expression of an ERK2 siRNA. These data suggest that BMP-2/p38 pathway links to the early phase, whereas ERK1/2 pathway is associated with the later phase in osthole-mediated differentiation of osteoblast cells. In this study, we demonstrate that osthole is a promising agent for treating osteoporosis.

The mechanism of ellipticine-induced apoptosis and cell cycle arrest in human breast MCF-7 cancer cells.

Ellipticine, a cytotoxic plant alkaloid, is known to inhibit topoisomerase II. Here, we first report the molecular mechanism of ellipticine's apoptotic action in human breast MCF-7 cancer cells. Treatment of cells with ellipticine resulted in inhibition of growth, and G2/M phase arrest of the cell cycle. This effect was associated with a marked increase in the protein expression of p53 and, p21/WAF1 and KIP1/p27, but not of WAF1/p21. Ellipticine treatment increased the expression of Fas/APO-1 and its ligands, mFas ligand and sFas ligand, and subsequent activation of caspase-8. The mitochondrial apoptotic pathway amplified the Fas/Fas ligand death receptor pathway by Bid interaction. This effect was found to result in a significant increase in activation of caspase-9. Taken together, we have concluded that the molecular mechanisms during ellipticine-mediated growth inhibition and induction of apoptosis in MCF-7 cells were due to (1) cell cycle arrest and induction of apoptosis, (2) induction of p53 and KIP1/p27 expression, (3) triggering of Fas/Fas ligand pathway, (4) disruption of mitochondrial function, and (5) the apoptotic signaling was amplified by cross-talk between Fas death receptor and mitochondrial apoptotic pathway.

Antioxidant activity of Cinnamomum cassia.

The antioxidant activities of hot water extracts (HWECC) and ethanol extracts (EECC) from the dry bark of Cinnamomum cassia Presl were evaluated in this study. Results showed that at 1.0 mg/mL, the ethanol extracts of C. cassia (96.30%) exhibited a greater inhibition than the alpha-tocopherol (93.74%) on FeCl(2)-ascorbic acid induced lipid peroxidation of rat liver homogenate in vitro. From 0.05 to 1.0 mg/mL, the EECC demonstrated the highest superoxide anions scavenging activity and the strongest anti-superoxide formation activity (p < 0.05). The same extract also showed an excellent antioxidant activity in enzymatic and nonenzymatic liver tissue oxidative systems. EECC revealed the strongest antioxidant activity followed by alpha-tocopherol and HWECC. Compared to alpha-tocopherol, the IC(50) values of EECC were found to be lower in thiobarbituric acid test (IC(50) = 0.24 mg/mL vs 0.37 mg/mL), in cytochrome c test (IC(50) = 0.16 mg/mL vs 0.27 mg/mL) and in xanthine oxidase inhibition test (IC(50) = 0.09 mg/mL vs 0.19 mg/mL). The present study concludes that EECC could be used as a good source of antioxidant in the dietary supplement.