Genetic analysis of developmental traits associated with enhanced winter survival in autumn-seeded rye (Secale cereale L.).

The abstract of this item is unavailable due to an embargo

Genetic analysis of developmental traits associated with enhanced winter survival in autumn-seeded rye (Secale cereale L.).

The abstract of this item is unavailable due to an embargo

Isolation and characterization of acetylcholinestrase inhibitors from Aquilaria species for treatment of Alzhimer Disease (AD) / Hirbod Bahrani

Aquilaria subintegra locally known as “Gaharu” belongs to the Thymelaeceae family. Its leaves have been claimed by Malay traditional practitioner in Malaysia were effective for the treatment of Alzheimer’s disease (AD). However, there is no scientific evidence available to support these claims. The AD is one of the most common neurodegenerative disorders and it causes dementia with aging. The acetylcholesterase (AChE) inhibitors were used for the treatment of AD. In this research the leaves and stem chloroform extract of A. subintegra were tested for AChE inhibitory activity. The TLC results of the leaves and stem of A. subintegra showed the presence of phenols, flavonoids, terpenoids, and alkaloids compounds in the extracts. The total phenol contents in the leaves and stem chloroform extracts were 164mgGAE/g and 210mgGAE/g respectively. Whereas, the total flavonoids contents in leaves and stem chloroform extracts were 414mgQE/g and 645mgQE/g respectively. The analysis of the stem chloroform extracts with LCMS/MS displayed that it contain kaempferol 3,4,7-trimethyl ether. The AChE inhibitory activity of leaves, stem chloroform extracts and kaempferol were 80%, 93% and 85.8% respectively. Brine Shrimp Lethality Assay (BSLA) exhibited LC50=11.64μg/ml of leaves chloroform extract, LC50=3.11μg/ml of stem chloroform extract, and LC50=9.59μg/ml for kaempferol. The response of leaves, stem chloroform extracts and kaempferol were perceived in Radial Arm Maze (RAM) through oral gavage to ICR male and female mice with impaired memory by valium. Administration of kaempferol to the mice significantly reduced the number of repeated entries to arms of maze in male and females rats. In conclusion, it could be postulated that the inhibition of AChE by leaves and stem chloroform extracts of A. subintegra could be due to the presence of kaempferol. Thus, it is safe to be used as a natural AChE inhibitor instead of berberine for the treatment of AD

The Relationships between Plant Developmental Traits and Winter Field Survival in Rye (Secale cereale L.)

Overwintering cereals accumulate low temperature tolerance (LTT) during cold acclimation in the autumn. Simultaneously, the plants adjust to the colder season by making developmental changes at the shoot apical meristem. These processes lead to higher winter hardiness in winter rye varieties (Secale cereale L.) adapted to Northern latitudes as compared to other cereal crops. To dissect the winter-hardiness trait in rye, a panel of 96 genotypes of different origins and growth habits was assessed for winter field survival (WFS), LTT, and six developmental traits. Best Linear Unbiased Estimates for WFS determined from five field trials correlated strongly with LTT (r = 0.90, p < 0.001); thus, cold acclimation efficiency was the major contributor to WFS. WFS also correlated strongly (p < 0.001) with final leaf number (r = 0.80), prostrate growth habit (r = 0.61), plant height (r = 0.34), but showed weaker associations with top internode length (r = 0.30, p < 0.01) and days to anthesis (r = 0.25, p < 0.05). The heritability estimates (h2) for WFS-associated traits ranged from 0.45 (prostrate growth habit) to 0.81 (final leaf number) and were overall higher than for WFS (h2 = 0.48). All developmental traits associated with WFS and LTT are postulated to be regulated by phytohormone levels at shoot apical meristem

The Relationships between Plant Developmental Traits and Winter Field Survival in Rye (<i>Secale cereale</i> L.)

Overwintering cereals accumulate low temperature tolerance (LTT) during cold acclimation in the autumn. Simultaneously, the plants adjust to the colder season by making developmental changes at the shoot apical meristem. These processes lead to higher winter hardiness in winter rye varieties (Secale cereale L.) adapted to Northern latitudes as compared to other cereal crops. To dissect the winter-hardiness trait in rye, a panel of 96 genotypes of different origins and growth habits was assessed for winter field survival (WFS), LTT, and six developmental traits. Best Linear Unbiased Estimates for WFS determined from five field trials correlated strongly with LTT (r = 0.90, p p p 0.01) and days to anthesis (r = 0.25, p 0.05). The heritability estimates (h2) for WFS-associated traits ranged from 0.45 (prostrate growth habit) to 0.81 (final leaf number) and were overall higher than for WFS (h2 = 0.48). All developmental traits associated with WFS and LTT are postulated to be regulated by phytohormone levels at shoot apical meristem

Fusion of Protegrin-1 and Plectasin to MAP30 Shows Significant Inhibition Activity against Dengue Virus Replication

<div><p>Dengue virus (DENV) broadly disseminates in tropical and sub-tropical countries and there are no vaccine or anti-dengue drugs available. DENV outbreaks cause serious economic burden due to infection complications that requires special medical care and hospitalization. This study presents a new strategy for inexpensive production of anti-DENV peptide-fusion protein to prevent and/or treat DENV infection. Antiviral cationic peptides protegrin-1 (PG1) and plectasin (PLSN) were fused with MAP30 protein to produce recombinant antiviral peptide-fusion protein (PG1-MAP30-PLSN) as inclusion bodies in <i>E. coli</i>. High yield production of PG1-MAP30-PLSN protein was achieved by solubilization of inclusion bodies in alkaline buffer followed by the application of appropriate refolding techniques. Antiviral PG1-MAP30-PLSN protein considerably inhibited DENV protease (NS2B-NS3pro) with half-maximal inhibitory concentration (IC₅₀) 0.5±0.1 μM. The real-time proliferation assay (RTCA) and the end-point proliferation assay (MTT assay) showed that the maximal-nontoxic dose of the peptide-fusion protein against Vero cells is approximately 0.67±0.2 μM. The cell-based assays showed considerable inhibition of the peptide-fusion protein against binding and proliferating stages of DENV2 into the target cells. The peptide-fusion protein protected DENV2-challeged mice with 100% of survival at the dose of 50 mg/kg. In conclusion, producing recombinant antiviral peptide-fusion protein by combining short antiviral peptide with a central protein owning similar activity could be useful to minimize the overall cost of short peptide production and take advantage of its synergistic antiviral activities.</p></div

The percentage of survival of DENV2-challenged mice administered with peptide-fusion protein.

<p>The lethal dose of the peptide- fusion protein that kills 50 percent of the animals (LD₅₀) was evaluated to be more than 50 mg/kg. Then, four groups of the animals (n = 6) were inoculated with 4×10³ plaque-forming units (PFU) of the purified DENV2 (DENV2-isolate Malaysia M2, GenBank Toxonomy No.: 11062) and treated with 12.5, 25 and 50 mg/kg of the peptide-fusion protein by intraperitoneal administration; while the forth group was treated with PBS as a mock-administrated group. (A), (B) and (C) The survival percentage of the animals that were administrated with 12.5, 25 and 50 mg/kg of the peptide-fusion protein respectively, compared with the mock-administrated group. Kaplan- Meier analysis was performed to generate survival curves with Prism software 5.01 (GraphPad Software, San Diego, CA).</p

Evaluation of peptide cytotoxicity.

<p>Toxicity was measured to determine the maximal non-toxic dose test (MNTD) value of the inhibitory peptides, which is the highest concentration of the peptide that causes minimal toxic effects on the cells. This assay was carried out by seeding Vero cells at 1×10⁴ cells/well in triplicates and treated with increased concentrations of antiviral peptides. The cell culture was analyzed after 72 h using Non-Radioactive Cell Proliferation assay. In this study, the peptide concentration that showed 90% and above of the cell viability was considered as the MNTD value, assuming that approximately 90% of the cells were healthy.</p

Evaluation of the peptides antiviral activities using plaque formation assay.

<p>(A) Virus load expressed as plaque forming units per ml (p.f.u/ml) was significantly reduced after treatment with all the peptides compared to untreated cells. The peptide-fusion protein (PG1-MAP30-PLSN) showed the highest inhibition potential compared with the other peptides. (B) Plaque formation assay shows the reduction of plaque generation after the treatment of the infected cells with the peptides. (Two-way ANOVA with Bonferroni post-test, p<0.001).</p

Evaluation of the peptides antiviral activities using qRT-PCR.

<p>Viral RNA levels were quantified by qRT-PCR to determine the level of viral copy number after the treatment of DENV2-infected cells with the antiviral peptides. (A) Viral copy number was reduced after the treatment with the peptides compared with untreated control. The peptide-fusion protein (PG1-MAP30-PLSN) showed the highest reduction in the viral copy number amongst the other peptides in a time-dependent manner. (B) Inhibition of virus binding to the target cells that led to reduce dengue virus copy number after the treatment with the peptides. The peptide-fusion protein potentially inhibited virus binding to the target cell by considerable reduction in the viral copy number compared with untreated control. However, there was insignificant (p>0.05) effect of MAP30 on virus binding to the target cell. PG1 and PLSN similarly inhibited virus binding at significant levels. Results are expressed as mean ± SD from a representative experiment performed in quadruple experiments. (Two-way ANOVA with <u>Bonferroni</u> post-test, p<0.001).</p