Developing a Protocol for an Environmentally Preferable Purchasing Plus (Epp+) Program at Nasa Glenn Research Center

EPP+ or Environmentally Preferable Purchasing Plus is a Environmentally Preferable Purchasing program with incorporated a unique Life Cycle Assessment (LCA) scoring system which implemented at NASA, Glenn Research Center. A protocol has been developed to aid in selecting among products based on principles of Environmentally Preferable Purchasing (EPP) and Life Cycle Assessment (LCA). The protocol has been developed for use at NASA Glenn Research Center (GRC) but is intended to benefit anyone wishing to make sustainable purchasing decision. The protocol was designed as a web-based application which evaluates the life cycle factors, affirmative procurements, price, and performance of products that impact or benefit the environment and human health. There were 2 parts in the development of the EPP+ protocol. The first part was the development of product scoring system guidelines. All products are to be scored using a numerical rating system for each value. A numeric-based rating system has been developed by applying the priorities of the environmental protection and management policies for the purchase of products for GRC. These rating are then multiplied by a weighting factor which is a numeric value between 0 and 1.0. During the product evaluation process, weighting factors are again set according to the importance and priorities set by the Environmental Management Office (EMO). Second part was the development of the EPP+ computer software or protocol (ASP.net format). This protocol produces a score for a product based on a series of ratings and weighting factors given by the expert user. Following the same sequence of operation, various products can be scored and compared, and a list can be prepared for the products that qualify as environmentally preferable products. The lists of products are intended to be offered to contractors and other GRC personnel for their applications. Since the product score is a numeric value, it is easy for a non-expert person to understand and compare with some other products, rath

Comparison of glucose derivatives effects on cartilage degradation

<p>Abstract</p> <p>Background</p> <p>Glucosamine (GlcN) is a well-recognized candidate for treatment of osteoarthritis. However, it is currently used in derivative forms, such as glucosamine-hydrochloride (GlcN-HCl) or glucosamine sulfate (GlcN-S). However, the molecular mode of action remains unclear. In this study, we compared the effects of Glucose (Glc), Glucuronic acid (GlcA), Glucosamine hydrochloride (GlcN-HCl) and Glucosamine sulfate (GlcN-S) on cartilage degradation.</p> <p>Methods</p> <p>Porcine cartilage explants were co-cultured with recombinant human IL-1β and each tested substance for 3 days. HA, s-GAG and MMP-2 releases to media were measured using ELISA, dye-binding assay and gelatin zymography, respectively. Similar studies were performed in a human articular chondrocytes (HAC) monolayer culture, where cells were co-treated with IL-1β and each reagent for 24 hours. Subsequently, cells were harvested and gene expression measured using RT-PCR. All experiments were carried out in triplicate. Student's t-tests were used for statistical analysis.</p> <p>Results</p> <p>In cartilage explants treated with IL-1β, GlcN-S had the highest chondroprotective activity of all four chemicals as shown by the inhibition of HA, s-GAG and MMP-2 released from cartilage. The anabolic (aggrecan core protein; AGG, SOX9) and catabolic (MMP-3, -13) genes in HACs treated with IL-1β and with/without chemicals were studied using RT-PCR. It was found that, GlcN-HCl and GlcN-S could reduce the expression of both MMP-3 and -13 genes. The IL-1β induced-MMP-13 gene expression was decreased maximally by GlcN-S, while the reduction of induced-MMP-3 gene expression was greatest with GlcN-HCl. Glc and GlcA reversed the effect of IL-1β on the expression of AGG and SOX9, but other substances had no effect.</p> <p>Conclusion</p> <p>This study shows that glucosamine derivatives can alter anabolic and catabolic processes in HACs induced by IL-1β. GlcN-S and GluN-HCl decreased induced MMP-3 and -13 expressions, while Glc and GlcA increased reduced-AGG and SOX9 expression. The chondroprotective study using porcine cartilage explant showed that GlcN-S had the strongest effect.</p

Methoxyflavone derivatives modulate the effect of TRAIL-induced apoptosis in human leukemic cell lines

<p>Abstract</p> <p>Background</p> <p>Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in various tumor cells, but does not affect normal cells or human leukemic cells, such as MOLT-4 and U937 cells, which are relatively resistant to TRAIL. Three flavonoids extracted from the rhizome of <it>K. parviflora </it>were 5,7-dimethoxyflavone (DMF), 5,7,4'-trimethoxyflavone (TMF) and 3,5,7,3',4'-pentamethoxyflavone (PMF), and synthetic flavonoids including 5-methoxyflavone (5-MF) and 2'-methoxyflavone (2"-MF) were chosen for testing in this study. The aims of this study were to examine whether the treatment of TRAIL-resistant leukemia MOLT-4 and U937 cells, with methoxyflavone derivatives could enhance the apoptotic response and to identify the mechanism involved.</p> <p>Methods</p> <p>The cytotoxic effect of methoxyflavone (MF) derivatives in MOLT-4, U937 and peripheral blood mononuclear cells (PBMCs) was analyzed by the MTT assay. The induction of apoptosis and the reduction of mitochondrial transmembrane potential (ΔΨm) after staining with annexin V FITC and propidium iodide (PI), and 3,3'-dihexyloxacarbocyanine iodide (DiOC₆), respectively, were performed using flow cytometry. ROS production was determined by staining with 2',7'-dichlorofluorescin diacetate and processed with a flow cytometer. DR4, DR5, cFLIP, Mcl-1, BAX and Bid expression were demonstrated by immunoblotting. Caspase-8 and -3 activities were determined by using IETD-AFC and DEVD-AFC substrates and the fluorescence intensity was measured.</p> <p>Results</p> <p>All methoxyflavone derivatives were cytotoxic to MOLT-4, U937 cells and PBMCs, except DMF, TMF and PMF were not toxic to PBMCs. All MF derivatives induced human leukemic MOLT-4 cell apoptosis, but not in U937 cells. Percentage of MOLT-4 cells with (ΔΨm) was increased when treated with DMF, TMF, PMF, 5-MF and 2'-MF in the presence of TRAIL. 5-MF and 2'-MF enhanced TRAIL-induced apoptosis through the up-regulation of both DRs and the down-regulation of cFLIP and Mcl-1. Bid was cleaved and BAX was up-regulated, followed by the activation of caspase-8 and -3. Oxidative stress was also increased. 2'-MF gave the same result compared with 5-MF but with a less effect.</p> <p>Conclusion</p> <p>Methoxyflavone derivatives enhanced TRAIL-induced apoptosis in human leukemic MOLT-4 cells through the death receptors and mitochondrial pathways.</p

Sesamin stimulates osteoblast differentiation through p38 and ERK1/2 MAPK signaling pathways

BACKGROUND: Osteoporosis is a worldwide health problem predominantly affecting post-menopausal women. Therapies aimed at increasing bone mass in osteoporetic patients lag behind comparable investigation of therapeutic strategies focusing on the bone resorption process. Sesamin, a major lignan compound found in Sesamun indicum Linn., has a variety of pharmacological effects, though its activity on bone cell function is unclear. Herein we examine the effect of this lignan on osteoblast differentiation and function. METHOD: Cell cytotoxicity and proliferative in hFOB1.19 were examined by MTT and alamar blue assay up to 96 h of treatment. Gene expression of COL1, ALP, BMP-2, Runx2, OC, RANKL and OPG were detected after 24 h of sesamin treatment. ALP activity was measured at day 7, 14 and 21 of cultured. For mineralized assay, ADSCs were cultured in the presence of osteogenic media supplement with or without sesamin for 21 days and then stained with Alizarin Red S. MAPK signaling pathway activation was observed by using western blotting. RESULTS: Sesamin promoted the gene expression of COL1, ALP, OCN, BMP-2 and Runx2 in hFOB1.19. On the other hand, sesamin was able to up-regulate OPG and down-regulate RANKL gene expression. ALP activity also significantly increased after sesamin treatment. Interestingly, sesamin induced formation of mineralized nodules in adipose derived stem cells (ADSCs) as observed by Alizarin Red S staining; this implies that sesamin has anabolic effects both on progenitor and committed cell stages of osteoblasts. Western blotting data showed that sesamin activated phosphorylation of p38 and ERK1/2 in hFOB1.19. CONCLUSIONS: The data suggest that sesamin has the ability to trigger osteoblast differentiation by activation of the p38 and ERK MAPK signaling pathway and possibly indirectly regulate osteoclast development via the expression of OPG and RANKL in osteoblasts. Therefore, sesamin may be a promising phytochemical that could be developed for supplementation of osteoporotic therapy

Anti-inflammatory effect of methanol extracts of hemp leaf in IL-1β-induced synovitis

Purpose: To examine the effectiveness of some hemp (Canabis sativa) leaf extracts as an antiinflammatory agent on synovitis in vitro.Methods: Synovial fibroblast cell line SW982 was induced with 5 ng/mL of interleukin 1-beta (IL-1β) to trigger cellular inflammation. The cells were then treated with prepared extracts of hemp (Canabis sativa) leaf originating from three different cultivation sites with varying proportions of terpenoids and cannabinoids, especially cannabidiol (CBD) and tetrahydrocannabinol (THC). Nitric oxide (NO) and prostaglandin E2 (PGE2) production as well as expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and IL-1β genes were determined.Results: All hemp extracts reduced the production of NO and PGE2 in a dose-dependent manner. The expressions of iNOS, COX-2 and IL-1β genes were significantly decreased by the hemp extracts. This effect was likely related to the amount of sesquiterpenoids and THC. The extract from Huai Hoi (HH) cultivar showed the most promising results for further studies.Conclusion: The extracts of hemp leaf substantially reduces the level of biomarkers for inflammation in vitro. Therefore, the extracts have a potential application as an inflammatory counteractant in synovitis.Keywords: Canabis sativa, Hemp, Synovitis, Cannabinoids, Nitric oxide, Interleukin-1 bet

Targeting human osteoarthritic chondrocytes with ligand directed bacteriophage-based particles

Osteoarthritis (OA) is a degenerative joint disease characterized by progressive deterioration and loss of articular cartilage. There is currently no treatment to reverse the onset of OA. Thus, we developed a targeted delivery strategy to transfer genes into primary human chondrocytes as a proof-of-concept study. We displayed a chondrocyte-affinity peptide (CAP) on the pIII minor coat protein of the M13 filamentous bacteriophage (phage)-based particle carrying a mammalian transgene cassette under cytomegalovirus CMV promoter and inverted terminal repeats (ITRs) cis elements of adeno-associated virus serotype 2 (AAV-2). Primary human articular chondrocytes (HACs) were used as an in vitro model, and the selectivity and binding properties of the CAP ligand in relation to the pathogenic conditions of HACs were characterized. We found that the CAP ligand is highly selective toward pathogenic HACs. Furthermore, the stability, cytotoxicity, and gene delivery efficacy of the CAP-displaying phage (CAP.Phage) were evaluated. We found that the phage particle is stable under a wide range of temperatures and pH values, while showing no cytotoxicity to HACs. Importantly, the CAP.Phage particle, carrying a secreted luciferase (Lucia) reporter gene, efficiently and selectively delivered transgene expression to HACs. In summary, it was found that the CAP ligand preferably binds to pathogenic chondrocytes, and the CAP.Phage particle successfully targets and delivers transgene to HACs

Antimutagenicity and DT-diaphorase inducing activity of Gynostemma pentaphyllum Makino extract

The hotwater extract of the herbal tea, Gynostemma pentaphyllum Makino, was not found tobemutagenicinSalmonellamutationassaywithorwithoutmetabolicactivation.However, the extract had both DT-diaphorase inducing activity in the murine hepatoma (Hepa1c1c7) cell line and antimutagenic properties towards chemical-induced mutation in Salmonella typhimurium strains TA98 and TA100. Mutagenicity of aflatoxinB1(AFB1),2-amino-6-methyldipyrido [1, 2-a : 3’, 2’, 3-d] imidazole (Glu-P-1), 2-aminodipyrido [1, 2-a:3’,2’,3-d]imidazole(Glu-P-2),2-amino- 1, 4-dimethyl-5H-pyrido [4, 3-b] indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido [4, 3-b] indole (Trp-P-2), 2-amino-3-methylimidazo [4, 5-f] quinoline (IQ) and Benzo [a] pyrene (B[a]P)was inhibited by the extract of Gynostemma pentaphyllum Makino in a dose-dependent manner, but no effect was found on the mutagenic activity of 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2). However, the extract enhanced themutagenicity induced by 2-aminoanthracene (2AA), and N-methyl-N’-nitro-N-nitrosoguanidine (MNNG)

ACTIVE COMPOUND OF ZINGIBER CASSUMUNAR ROXB. DOWN-REGULATES THE EXPRESSION OF GENES INVOLVED IN JOINT EROSION IN A HUMAN SYNOVIAL FIBROBLAST CELL LINE

Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovium. It is involved in up-regulation of pro-inflammatory cytokines and matrix metalloproteinases (MMPs), resulting in joint inflammation and erosion. Zingiber cassumunar Roxb. has long been used to reduce joint pain and inflammation. This study aimed to investigate the inhibitory activities of an active compound of Z. cassumunar, (E)-4-(3',4'-dimethoxyphenyl)but-3-en-1-ol (compound D), against cytokine-induced up-regulation of catabolic genes involved in cartilage degradation in RA. Synovial fibroblast cell line, SW982, was cultured in media containing interleukin-1ß (IL-1ß), in the presence or absence of compound D at the concentration range of 1 to 100 µM. After 24 hours, the cells were analyzed for the expressions of MMPs, IL-1ß and interleukin-1ß-converting enzyme (ICE) by RT-PCR. MMPs activities in the culture media were analyzed by zymographic techniques. Dexamethasone was used as the positive control. It was found that compound D at the concentration of 10 - 100 M significantly decreased the mRNA expressions of MMP-1, -2, -3, and -13 which was induced by IL-1ß (