. NO activates soluble guanylate cyclase (sGC) by binding to the heme site on the protein, inducing a conformational change (Bellamy et al., 2002). sGC in turn modulates the transformation of guanosine triphosphate (GTP) into cyclic guanosine monophosphate (cGMP) (Garthwaite et al., 1988). cGMP acts as a second messenger and its production activates protein kinases (PKG), phosphodiesterases (PDE III) and cGMP-dependent kinase II (cGKII). An increase in AMPA-Rs has been reported to occur through cGKII binding to the CTD of GluR1. Blocking cGKII activity led to a reduction in LTP in hippocampal slices (Serulle et al., 2007). Besides, the NO/cGMP/PKG pathway has been shown to mediate actin depolymerisation and synaptic retraction through ROCK signalling in motor neurons (Sunico et al., 2010). Moreover, NO can regulate the Akt kinase signalling pathway and CREB (Calabrese et al., 2007). NO can also directly modify proteins through S-nitrosylation of MT associated protein (MAP) that has been shown to lead to growth cone collapse and axonal actin reconfiguration (Stroissnigg et al., 2007; Hardingham et al., 2013). For example, GluR1 AMPA-R subunit NO-mediated-S-nitrosylation has been linked to an increase in AMPA-R facilitating LTP and synaptic plasticity (Selvakumar et al., 2013).
M any investigations have recently highlighted the importance of the gut microbi- ota in the onset and progression of a number of human diseases, including gastrointestinal disorders (1, 2), inﬂammatory diseases (3), respiratory tract infections (4), and allergies (5). The functional impact of commensal gut microorganisms depends on their ability to survive in the gastrointestinal tract, adhere to epithelial mucus, and obtain energy from nondigestible dietary substrates and host mucosal secretions (6). More than half of all proteins in nature have been estimated to be glycosylated through O-glycosidic or N-glycosidic bonds (7). O-Glycans are linked to a serine or threonine residue via an N-acetylgalactosamine, which is elongated by additional sugars (8). N-Glycosylation is a common modiﬁcation of extracellular membrane proteins present at the gastrointestinal epithelium, the secreted proteins of human breast milk, and many dietary proteins (7, 9–11). N-Linked glycans are attached via the core N,N=- diacetylchitobiose disaccharide (ChbNAc) (GlcNAc- ␤ 1,4-GlcNAc) to an asparagine resi- due of proteins containing the Asn-Xxx-Ser/Thr (with Xxx being any amino acid except Pro) motif (12). In N-glycans from mammals, the inner GlcNAc moiety bound to Asn is often fucosylated through an ␣ 1,6 linkage, named core fucose. Protein N-glycosylation plays a crucial role in a variety of cellular processes, such as cell adhesion (13), immune pathway signaling (14), and bacterial recognition (15). Some intestinal microorganisms have the ability to process the carbohydrate moieties of N-glycosylated proteins (16) so that the type, abundance, and location of these glycans contribute to shaping the composition and distribution of the gut microbiota (17). Some bacterial pathogens possess endo- ␤ -N-acetylglucosaminidase enzymes that cleave the ␤ -1,4 linkage of the core ChbNAc present in all N-glycoproteins, releasing the N-glycan moiety (18). The activities of these enzymes have been
Materials and methods: In the rat model of acute ocular hypertension (AOH), intraocular pressure was increased to 110 mmHg for 60 minutes. Animals were divided into four groups: sham operation (Ctrl), AOH, AOH + phosphate-buffered saline (PBS), and AOH + AAA. Cell apoptosis in the ganglion cell layer was detected with the terminal deoxynucleotidyl transferase- mediated uridine 5′-triphosphate-biotin nick end labeling (TUNEL) assay, and retinal ganglion cells (RGCs) immunostained with Thy-1 were counted. Müller cell activation was detected using immunostaining with glutamine synthetase and glial fibrillary acidic protein. Tumor necrosis factor-α (TNF-α) was examined using Western blot.
The cost of free fatty acid as exogenous substrate in cul- ture is too high to justify their use in EPA production in M. alpina. To reduce costs, triglycerides can be used as a substrate for EPA production in Ma-MpFADS6 trans- formants. Peony seed oil (PSO) was selected as a suitable exogenous substrate, because it contains 46 % ALA and 26 % LA (Additional file 3: Fig. S3). Our results revealed that the DCW and TFA of all strains were not signifi- cantly altered in cultures after PSO addition (Fig. 4a, b). When PSO concentration was increased, the level of AA was not significantly raised, but it was sharply reduced when the PSO concentration reached 0.2 % (Fig. 4c). Similar results were obtained after supplementing cul- tures with 0.4 and 0.6 % PSO, data not shown. The Ma- MpFADS6 transformants presented a noticeably higher EPA production with increasing PSO concentrations, with a maximum level of 149.3 ± 7.8 mg/L with 0.1 % PSO, which represents a 3.5-fold increase relative to 42.5 ± 2.7 mg/L in WT M. alpina (Fig. 4d). After sup- plementing cultures with 0.025, 0.05 and 0.1 % PSO in all Ma-MpFADS6 transformants, about half of the intracel- lular ALA was converted to EPA by MpFADS6, followed by further fatty acid elongation and desaturation steps (Fig. 4e). However, after supplementation with 0.2 % PSO, the level of intracellular ALA in Ma-MpFADS6 transformants was approximately equivalent among all strains (Fig. 4e), and the level of EPA was reduced to half
solani . While γ -aminobutyric acid (GABA) as another derivative can accumulate proline, reduce chilling injury and activate the defense response of peach fruit during long-term cold storage [10, 11]. As a structural isomers of AIB, β -aminoisobutyric acid (BAIB) has a key metabolic role in enhancing fatty acid oxidation by converting the cells of white adipose tissue into brown fat and turning off the energy stores, which may help to prevent diet-induced obesity and related metabolic disorders . Unfortu- nately, information is extremely limited on roles of BAIB in harvested horticultural crops. Thus, it can be expected AIB and BAIB as postharvest alternatives for longan fruit by inducing defense activity and delaying senescence.
tical neurons death via the PI3K/Akt cell survival pathway [22,23]. A binding mechanism has been established in which α-synuclein and Tau bind to DNA in a specific con- formation that causes transformational transition and stabilization of Z-DNA conformation [24,25]. Tau and α- synuclein bind to the B-DNA conformation and alter its form by increasing the melting temperature and decreas- ing the EtBr molecules bound per base pair in the B form of DNA. The altered B form is favored in DNA stability and is a probable intermediate to form the favoring Z-DNA conformation, which are inferred to have a sig- nificant role in gene expression in neurodegeneration . Furthermore, it was found that aggregated α-synuclein can directly cross-seed tau fibrillization. Preformed α- synuclein fibrils were assembled from recombinant pro- tein in vitro (primary neurons) and in vivo (transgenic mice). It was concluded that there were at least two dis- tinct strains of synthetic α-synuclein fibrils with differ- ences in the efficiency of cross-seeding tau fibrilization.
The disappointing clinical outcome of Val-boroPro does not exclude the potential role of FAP α’s proteolytic activity in tumor invasion and metastasis, or that an inhibitor antibody may be a potent therapeutic target. An inhibitory scFv antibody, named E3 was identified, which competitively inhibits FAP α function . This scFv antibody with high affinity and enhanced inhibitory effects on FAP α enzyme activity, seems very likely to be exploited as a tool for the treatment of FAP α driven tumors. Studies to evaluate the effects of scFv antibody deserve more exploration and further characterization to confirm previous findings. In one study, human scFvs were transformed into bivalent minibodies of completely human origin, which worked far better than murine or humanized antibody derivatives. Thus, the successful use of mini-antibodies in immunohistology for a variety of carcinomas is encouraging for in vivo diagnostic and tumor-targeting studies . Combinatorial strategies addressing the two key issues of cancer immunotherapy (ie. targeting the tumor cells and modulating the T-cell response), the production of a bio-specific single chain
We are grateful to Stephanie Fowler and Kalin Mayberry (Depart- ment of Hematology, St. Jude Children’s Research Hospital) for mouse studies; Victoria Frohlich, Sharon Frase, Linda Horn- er, and Randall Wakefield (St. Jude Cellular Imaging Shared Research Core); Sean Savage, Michael Anderson, and Joseph Emmons (St. Jude Veterinary and Pathology Core); Shao Youm- ing and Heath Richard (St. Jude Protein Production Core); Cecil- ia Western (St. Jude Vector Core); and Keith A. Laycock (St. Jude Department of Scientific Editing). We thank Huiying Li and Thomas L. Poulos (University of California, Irvine) for providing full-length recombinant eNOS protein (supported by NIH grant GM57353), for helpful discussions on eNOS biochemistry, and for reviewing this manuscript. We thank Gilles Truan (Institut National des Sciences Appliquées de Toulouse, Toulouse, France) for providing the CyB5 expression plasmid. We thank John Olson (Rice University, Houston, Texas, USA) for helpful discussions. This work was supported by NIH grants R01 DK61692 (to MJW) and R01 HL088554 (to BEI and MJW); an American Heart Asso- ciation Postdoctoral Fellowship (to CL); and the American Leba- nese Syrian Associated Charities (ALSAC).
in land as a major livelihood asset are discussed. The case for this discussion is the Wasa Amenfi West district in the Western region of Ghana. The research aimed to show that land as a livelihood asset is more than just a natural product; it is also socially constituted through the strategies of people to secure and deliver their livelihood assets. Qualitative research approach was considered in collecting data from farmers in the Wasa Amenfi West District. Thus, what is often considered livelihood strategies in the livelihood discourse are not adequately presented as a necessary condition through which resources become assets in the first place. The paper therefore demonstrates that strategies of people such as the migrant farmers’ group formation provide social, economic, and political utility for delivering land as key livelihood asset, such that access to and use of land cannot be possible without these strategies.To this end therefore, in a proper perspective, strategies of people should be considered as a necessary social constituent that complement the naturalconstituent of resources to deliver them as assets.
As the precursor of the ω-3 PUFA metabolic pathway, ALA is extremely important for the sufficient supply of long chain ω-3 fatty acids. Because mammals cannot synthesize ALA de novo, ALA must be included in the human diet, which is why ALA is called essential fatty acid. ALA is primarily found in plant source. The richest amounts of ALA from plant sources are flaxseed and camelina oils, which have 53 and 38% (in average) of ALA respectively. Unfortunately, flaxseed oil is con- sumed by only a limited population, whereas camelina oil is not commonly consumed as part of the diet . Vegetable oils are the common dietary source of ALA, but the ALA contents are relatively low. As examples, rapeseed and peanut oil contain 8.1 and 0.5% of ALA respectively . Recently, microalgae as potential sources of ω-3 PUFAs have raised a great interest. Microalgae are
In terms of application it seems necessary to thoroughly analyze the mechanisms of antioxidant actions of mela- tonin and to distinguish between effects observed at phar- macological or physiological concentrations. These considerations must not be restricted to the melatonin released from the pineal gland into the circulation and to the classic hepatic degradation route of 6-hydroxylation followed by conjugation. On the contrary, we would like to lay emphasis on the significance of tissue melatonin and the alternate oxidative pathways of catabolism lead- ing to different, biologically active products. The relation- ship between melatonin and nutrition will be discussed, with regard to the presence of the compound as a natural food constituent sometimes affecting circulating levels, to the post-prandial release of melatonin from the gastroin- testinal tract, and to interactions with other antioxidants present in food. Finally, a model of mitochondrial protec- tion is reviewed.
Constituent based Accent Prediction Constituent based Accent Prediction Christine H Nakatani A T & T L a b s R e s e a r c h 180 P a r k A v e n u e , F l o r h a m P a r k N J 0 7 9 3 2 0 9 7 1 , U S[.]
Tumor-draining inguinal lymph nodes were resected and pushed through a 70-μm nylon sieve (BD Bios- ciences Discovery Labware, Two Oaks, CA, USA) to produce a single cell suspension. The cells were then washed (300 × g for 7 minutes) and filtered through a 40- μ m nylon sieve (BD Biosciences). Spleens were resected and pushed through a 70- μ m nylon sieve to produce a single cell suspension. After red blood cell lysis, the cells were washed (300 × g for 7 minutes) and filtered through a 40-μm nylon sieve. To isolate tumor- infiltrating immune cells (TICs), tumors were resected and minced using a scalpel blade in a triple enzyme digestion mix containing 10 mg/mL collagenase type IV (Worthington Biochemical Corp., Lakewood, NJ, USA), 1 mg/mL hyaluronidase (Sigma-Aldrich, St. Louis, MO, USA), 200 μg/mL DNAse I (Roche Applied Sciences, Indianapolis, IN, USA) in Hanks ’ Balanced Salt Solution (Lonza). The tumors were then incubated with agitation (37°C for 45 minutes). After the addition of 10 mM ethylenediaminetetraacetic acid, the digestion product was incubated for another 15 minutes. Subsequently, the digested tissue was pushed sequentially through 70- μ m
To date, numerous solid acid catalysts have been developed, including zeolite , heteropolyacids , ion exchange polymer resins [8–10], carbon-based solid acids , and zinc lanthanum mixed oxides . These solid acid catalysts lead to a more facile process and a lower production cost for esterification reactions. However, the limited active sites of these heterogeneous solid acid catalysts prohibit their widespread application. In addition, their relatively low porosity is not favorable for the mass transfer of reactants by diffusion, which leads to a low reaction rate. As a result, the development of solid acid catalysts with a high concentration of acid sites that are easily accessible is highly desired.
Dry-column flash chromatography fractionation of the EO was performed in a sintered glass column (300 mm×32 mm) packed with silica gel (100 g Merck, <0.08 mm) using water pump vacuum. The column was equilibrated with 750 mL of the mobile phase toluene/ethyl acetate (70:30 volume ratio) solvent system. The EO (m = 812 mg) was diluted with the mobile phase (1:1) before applying to the column. The elution was isocratic and 25 fractions of around 10 mL were collected. After the HPTLC-direct bioautography analysis of EO, the position of the active component (isogeranic acid), i.e., its R F value was determined. Ordinary TLC chromatography of all of the fractions from dry flash chromatography revealed the frac- tions 7 and 8 which contained a spot of the same R F value which exhibited antibacterial acti- vity. These fractions, were joined, evaporated on the rotary vacuum evaporator (Ika-werke, RV10, Staufen, Germany) without heating, and further separated on the second dry-column flash chromatography packed with the silica gel (SiO 2 , <0.08 mm, Merck), with the same dimensions as the first one. The elution was gradient, starting with the toluene–ethyl acetate (98:2 volume ratio) and ending with the toluene–ethyl acetate (95:5 volume ratio). Forty fractions were collected and the volume of each fraction was 10 mL. The last two fractions, which contained an “active spot”, were grouped and evaporated on the rotary vacuum evapor- ator without heating, and the final separation was performed on the third dry-column flash chromatography in a sintered glass column (85 mm× 20 mm) packed with silica gel (SiO 2 ,