Salinity stress improves antioxidant potential by modulating physio-biochemical responses in Moringa oleifera Lam.

Moringa oleifera Lam. is a common edible plant, famous for several nutritional and therapeutic benefits. This study investigates the salt -induced modulations in plant growth, physio-biochemical responses, and antioxidant performance of M. oleifera grown under 0, 50, and 100 mM NaCl concentrations. Results showed that the plant effectively managed moderate salinity (50 mM NaCl) by maintaining succulence, weight ratios, and biomass allocation patterns of both shoot and root with minimal reduction in dry biomass. However, high salinity (100 mM NaCl) remarkably declined all growth parameters. The plant accumulated more Na+ and Cl-, while less K+ under salinity as compared to the control. Consequently, osmotic potentials of both root and leaf decreased under salinity, which was corroborated by the high amount of proline and soluble sugars. Increased level of H2O2 with significantly unchanged membrane fluidity indicating its role in perceiving and managing stress at moderate salinity. In addition, increased activities of superoxide dismutase, and catalase, with increased glutathione and flavonoid contents suggest an integrated participation of both enzymatic and non-enzymatic antioxidant components in regulating ROS. On the other hand, high salinity caused an outburst of ROS indicated by high H2O2, MDA, and electrolyte leakage. As a response, moringa drastically increased the activities of all antioxidant enzymes and contents of antioxidant molecules including ascorbic acid, glutathione, total phenols, and flavonoids with high radical scavenging and reducing power capacities. However, a considerable amount of energy was used in such management resulting in a significant growth reduction at 100 mM NaCl. This study suggests that moringa effectively resisted moderate salinity by modulating physio-biochemical attributes and effectively managing ion toxicity and oxidative stress. Salt stress also enhanced the medicinal potentials of moringa by increasing the contents of antioxidant compounds including ascorbic acid, glutathione, total phenols, and flavonoids and their resulting activities. It can be grown on degraded/ saline lands and biomass of this plant can be used for edible and medicinal purposes, besides providing other benefits in a global climate change scenario.

Effects of different conditions tested "in vitro" on the phosphorus runoff potential of livestock manure.

This study aimed to investigate the changes of swine and dairy manure characteristics during a long-term storage (150-180 days) under 4 °C, 20 °C, and 37 °C, sealed and unsealed conditions. Water extractable phosphorus (WEP) of both manures rapidly increased during the first 15-30 days and then decreased. At the end of the storage, the WEP reduction was 90%±3% and 71%±5% of the initial concentration for swine manure and dairy manure, respectively. Generally, unsealed storage and higher temperatures led to more WEP reduction. This study suggested that manure stored for less than 30 days had the highest P runoff potential, while a long-term manure storage reduced P runoff potential compared to freshly excreted manure.

Upgrading Solid Digestate from Anaerobic Digestion of Agricultural Waste as Performance Enhancer for Starch-Based Mulching Biofilm.

Developing a green and sustainable method to upgrade biogas wastes into high value-added products is attracting more and more public attention. The application of solid residues as a performance enhancer in the manufacture of biofilms is a prospective way to replace conventional plastic based on fossil fuel. In this work, solid digestates from the anaerobic digestion of agricultural wastes, such as straw, cattle and chicken manures, were pretreated by an ultrasonic thermo-alkaline treatment to remove the nonfunctional compositions and then incorporated in plasticized starch paste to prepare mulching biofilms by the solution casting method. The results indicated that solid digestate particles dispersed homogenously in the starch matrix and gradually aggregated under the action of a hydrogen bond, leading to a transformation of the composites to a high crystalline structure. Consequently, the composite biofilm showed a higher tensile strength, elastic modulus, glass transition temperature and degradation temperature compared to the pure starch-based film. The light, water and GHG (greenhouse gas) barrier properties of the biofilm were also reinforced by the addition of solid digestates, performing well in sustaining the soil quality and minimizing N2O or CH4 emissions. As such, recycling solid digestates into a biodegradable plastic substitute not only creates a new business opportunity by producing high-performance biofilms but also reduces the environmental risk caused by biogas waste and plastics pollution.

Periplocin induces apoptosis and inhibits inflammation in rheumatoid arthritis fibroblast-like synoviocytes via nuclear factor kappa B pathway.

Apoptotic resistance and excessive proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) stimulated by inflammation could lead to distal joint destruction and bone damage. Periplocin could promote apoptosis, resist proliferation, and reduce inflammation. However, the effect and mechanism toward periplocin in proliferation and inflammation of RA-FLSs remain unclear. The role of tumor necrosis factor (TNF)-α induced proliferation and expression of inflammatory cytokines in RA-FLSs was established. Our studies noted that cell viability of TNF-α-induced RA-FLSs was inhibited in periplocin treatment via dose-response, whereas cell apoptosis of RA-FLSs was triggered by dose-dependent effect of periplocin. Bcl-2 protein, one of the apoptotic regulators, was downregulated, while other regulators of apoptosis, including BAX, cleaved caspase-3, and cleaved caspase-9, were upregulated in RA-FLSs under periplocin treatment. In addition, periplocin decreased the TNF-α-induced mRNA and protein expression levels of interleukin (IL)-1ß and IL-6 in RA-FLSs in a dose-dependent way. Finally, the increased levels of phospho (p)-inhibitor of kappa B (IκBα)/IκBα and p-NF (nuclear factor)-κB/nuclear factor kappa B (NF-κB) ratio of RA-FLSs stimulated by TNF-α were decreased by periplocin treatment. Taken together, periplocin treatment decreased cell viability and cytokines expression and promoted cell apoptosis of TNF-α-induced RA-FLSs through inhibition of NF-κB signaling pathway, providing a potential therapeutic approach for RA.

Anti-CD40 mAb enhanced efficacy of anti-PD1 against osteosarcoma.

The overall survival rate of patients with osteosarcoma has remained stagnant at 15-30% for several decades. Although immunotherapy has revolutionized the oncology field, largely attributed to the success of immune-checkpoint blockade, the durability and efficacy of anti-PD1 (programmed cell death protein 1) mAb vary across different malignancies. Among the major reasons for tumor resistance to this immune checkpoint therapy is the absence of tumor-infiltrating cytotoxic T lymphocytes. However, the presence of intratumor exhausted PD1hi T cells also contributes to insensitivity to anti-PD1 treatment. In this study, we established the osteosarcoma mouse tumor model resistant to anti-PD1 mAb that harbored PD1hi T cells. Furthermore, flow cytometry analysis of tumor infiltrating leukocytes after treatment was used as a screening platform to identify agents that could re-sensitize T cells to anti-PD1 mAb. Results showed that anti-CD40 mAb treatment converted PD1hi T cells to PD1lo T cells, reversing phenotypic T cell exhaustion and sensitizing anti-PD1 refractory tumors to respond to anti-PD1 mAb. Results also showed that intratumor Treg presented with a less activated and attenuated suppressive phenotype after anti-CD40 mAb treatment. Our study provides proof of concept to systematically identify immune conditioning agents, which are able to convert PD1hi T cells to PD1lo T cells, with clinical implications in the treatment against refractory osteosarcoma to anti-PD1 mAb.

Effects of ANRIL variants on the risk of ischemic stroke: a meta-analysis.

Background: Several studies investigated the relationship between antisense non-coding RNA in the INK4 locus (ANRIL) variants and the risk of ischemic stroke (IS), yet whether ANRIL variants are associated with IS remain controversial. Therefore, we performed the present study to obtain a more conclusive result. Methods: Literature retrieval was conducted in PubMed, Medline and Embase. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. Results: Eighteen studies were enrolled for analyses. Pooled overall analyses showed that rs2383206 (recessive model: P=0.002, OR = 1.22, 95%CI 1.08-1.38; allele model: P=0.003, OR = 0.90, 95%CI 0.84-0.96) and rs10757274 (allele model: P=0.006, OR = 0.91, 95%CI 0.86-0.97) variants were significantly associated with an increased risk of IS. Further subgroup analyses by ethnicity revealed that rs2383206, rs10757274 and rs10757278 variants were all significantly correlated with an increased risk of IS in Asians. Additionally, rs10757278 polymorphism was also significantly correlated with an increased risk of IS in Caucasians. Conclusions: Our findings indicated that rs2383206, rs10757274 and rs10757278 variants may impact individual susceptibility to IS in Asians. Moreover, rs10757278 polymorphism may also impact individual susceptibility to IS in Caucasians.

Hsp90 inhibitor NMS-E973 exerts the anticancer effect against glioblastoma via induction of PUMA-mediated apoptosis.

BACKGROUND: Glioblastoma is one of the most aggressive and common malignancies of the central nervous system in humans. Owing to the correlation of high Hsp90 expression with prognosis and clinical pathology features of diverse types of cancer, targeting Hsp90 with small-molecule inhibitors has become a promising anticancer strategy. PURPOSE: In this study, we aimed to explore the possibility of anticancer effect of NMS-E973 in giloblastoma and elucidate the mechanism. METHODS: Cell based MTT assay and colony formation assay were used to detect cell viability. Apoptosis was analyzed by nuclear staining with Hoechst 33258 and Annexin V/propidium iodide staining followed by flow cytometry. Western-blot and RT-PCR were used to detect gene expression. Xenograft assay was used to explore the anticancer effect of NMS-E973 in vivo. RESULTS: We found that NMS-E973 induces apoptosis and inhibits cell growth in glioblastoma cells in cell culture and xenograft models. As a proapoptotic Bcl-2 member, PUMA was induced by NMS-E973 in a p53-dependent manner in glioblastoma in cell culture, thereby inducing apoptosis in glioblastoma cells. Furthermore, PUMA was induced by NMS-E973 treatment in xenograft tumors, and deficiency in PUMA significantly suppressed the antitumor effects of NMS-E973. CONCLUSION: Our study suggests that PUMA-mediated apoptosis is important for the therapeutic responses of NMS-E973. Induction of PUMA might be a potential biomarker for predicting NMS-E973 responses.

Arbuscular mycorrhizal fungi increase salt tolerance of apple seedlings.

BACKGROUND: Apple trees are often subject to severe salt stress in China as well as in the world that results in significant loss of apple production. Therefore this study was carried out to evaluate the response of apple seedlings inoculated with abuscular mycorrhizal fungi under 0, 2‰, 4‰ and 6‰ salinity stress levels and further to conclude the upper threshold of mycorrhizal salinity tolerance. RESULTS: The results shows that abuscular mycorrhizal fungi significantly increased the root length colonization of mycorrhizal apple plants with exposure time period to 0, 2‰ and 4‰ salinity levels as compared to non-mycorrhizal plants, however, percent root colonization reduced as saline stress increased. Salinity levels were found to negatively correlate with leaf relative turgidity, osmotic potential irrespective of non-mycorrhizal and mycorrhizal apple plants, but the decreased mycorrhizal leaf turgidity maintained relative normal values at 2‰ and 4‰ salt concentrations. Under salt stress condition, Cl- and Na+ concentrations clearly increased and K+ contents obviously decreased in non-mycorrhizal roots in comparison to mycorrhizal plants, this caused mycorrhizal plants had a relatively higher K+/Na+ ratio in root. In contrast to zero salinity level, although ascorbate peroxidase and catalase activities in non-inoculated and inoculated leaf improved under all saline levels, the extent of which these enzymes increased was greater in mycorrhizal than in non-mycorrhizal plants. The numbers of survived tree with non-mycorrhization were 40, 20 and 0 (i.e., 66.7%, 33.3% and 0) on the days of 30, 60 and 90 under 4‰ salinity, similarly in mycorrhization under 6‰ salinity 40, 30 and 0 (i.e., 66.7%, 50% and 0) respectively. CONCLUSION: These results suggest that 2‰ and 4‰ salt concentrations may be the upper thresholds of salinity tolerance in non-mycorrhizal and mycorrhizal apple plants, respectively.

[The structure-function relationship of thermostable beta-glycosidase from the thermophilic eubacterium Thermus nonproteolyticus HG102].

Beta-glycosidase (Tngly) from the thermophilic eubacterium Thermus nonproteolyticus HG102, which is a thermostable monomeric protein and adopts the (beta/alpha)8 barrel fold, is an excellent model system to be investigated for the thermostable mechanism, activity and substrate specificity. Here, based on the analysis of structural basis for thermostability of Tngly (Wang et al, 2003) and comparison of other proteins structure of homofamily, Glu164 and Glu338 may act as proton donor and nucleophile in the hydrolysis reaction respectively; proline located at N1 of alpha-helix and arginine which can form ion link may contribute to the thermostability. We aim to further identify the critical sites and the amino acid residue(s) responsible for the activity, the thermal stability and the substrate specificity. Mutations had been constructed by site-directed mutagenesis. They are Glu164Gln, Glu338Ala, Pro316Gly, Arg325Leu, Pro344Phe, Pro356Ala and Pro316Gly/Pro356Ala. All mutant proteins were purified to SDS-PAGE purity. Changes in the conformations were examined by means of CD. The Glu338Ala mutant showed no detectable hydrolysis activity, but can synthesize oligosaccharides, as expected for the residue acting as the nucleophile of the reaction. The Glu164 acts as the general acid/base catalyst in the hydrolysis reaction. Changes in stabilities of mutants compared with wild-type were determined by means of heat inactivity experiment. These results indicate that the amino acid residue of proline that is located at N1 positions of alpha-helix, and Arg325 that form salt bridge between alpha-helices 5 and alpha-helices 6, are the critical sites to protein thermostabilization.

Structural basis for thermostability of beta-glycosidase from the thermophilic eubacterium Thermus nonproteolyticus HG102.

The three-dimensional structure of a thermostable beta-glycosidase (Gly(Tn)) from the thermophilic eubacterium Thermus nonproteolyticus HG102 was determined at a resolution of 2.4 A. The core of the structure adopts the (betaalpha)(8) barrel fold. The sequence alignments and the positions of the two Glu residues in the active center indicate that Gly(Tn) belongs to the glycosyl hydrolases of retaining family 1. We have analyzed the structural features of Gly(Tn) related to the thermostability and compared its structure with those of other mesophilic glycosidases from plants, eubacteria, and hyperthermophilic enzymes from archaea. Several possible features contributing to the thermostability of Gly(Tn) were elucidated.

[Cloning and expression of a thermostable beta-glycosidase gene from Thermus nonproteolyticus HG102].

The gene coding for beta-glycosidase (EC3.2.1.21) from Thermus nonproteolyticus HG102 has been cloned and expressed in E. coli. The gene open reading frame was 1311 bp and it codes for 436 amino acids. The deduced amino acid sequence of the enzyme showed identity with members of glycosyl hydrolase family I. The enzyme had high content of hydrophobic amino acid (Ala 12.8%, Leu 10.9%), Arg(9.6%), Glu(9.4%) and Pro(8.0%), but low content Cys(0.45%) and Met (0.9%). From the alignment of enzyme amino acid sequence with other glycosyl hydrolase family I members, Glu164 and Glu338 were predicated as the proton donor and nucleophile group. The DNASTAR program was used to predict the secondary structure. According to the Chou-Fasman model, the enzyme has 41.4% of alpha-helics, 16.2%, beta-strands, 14.4%, beta-turns. 14 of the 35 Pro were located at the second sites of beta-turns. Hydrophobic interaction, ion bond, alpha-helics and Pro had important contribution to Tn-gly thermostability.