Contribution of phlA and some metabolites of fluorescent pseudomonads to antifungal activity

Fluorescent Pseudomonas species are an important group of PGPR that suppress fungal root and seedling disease by production of antifungal metabolites such as 2,4-diacetylphloroglucinol (2,4-DAPG), pyoluteorin, pyrolinitrin, siderophores and HCN. The compound 2,4-DAPG is a major determinant in biocontrol of plant pathogens. A 7.2 kbp chromosomal DNA region, carrying DAPG biosynthetic genes (phlA, phlC, phlB, phlD, phIE and phlF). Detecting the ph1 genes make them an ideal marker gene for 2,4-DAPG-producing fluorescent pseudomonad's. In this study we detected ph1A gene (that convert MAPG to 2,4-DAPG) using PCR assay with primers phlA-1r and phlA- f that enabled amplification of phlA sequences from fluorescent pseudomonad's from ARDRA group 1 and 3. We could detect phlA gene in P. fluorescens strains CHAO, Pf-44, Pf-1, Pf-2, Pf-3, Pf-17, Pf-62 and Pf-64, native isolates of Iran. The efficacy of this method for rapid assay characterizing rhizosphere population of 2,4-DAPG producing bacteria from soil of different area of Iran is in progress. We used a collection of 48 fluorescent pseudomonas strains in vitro, with known biological control activity against some soil born phytopathogenic fungi such as, Macrophomina phaseoli, Rhizoctonia solani Vericillium dahlia, Phytophthora nicotiana, Pythium spp. and Fusarium spp. and the potential to produce known secondary metabolites such as protease. Strains Pf-1, Pf-2, Pf-3, Pf-17, Pf-33 and Pf-44 showed the best antifungal activity against all fungi used in this study. Thirty-eight of 48 strains produced protease. The ability to rapidly characterize populations of 2,4-DAPG producers will greatly enhance our understanding of their role in the suppression of root disease.     

Nano reengineering of horseradish peroxidase with dendritic macromolecules for stability enhancement

A simple bio-conjugation procedure to surround a single horseradish peroxidase (HRP) enzyme molecule with dendritic polyester macromolecules (polyester-32-hydroxyl-1-carboxyl bis-MPA dendron, generation 5) was proposed. The characterization of resultant nanoparticles entitled HRP dendrozyme, was performed by transmission electron microscopy, dynamic light scattering, gel permeation chromatography and Fourier transform infrared spectroscopy. The results showed that HRP nanoparticles were spherical in shape and have an average size of 14 ± 2 nm in diameter. Furthermore, bio-conformational characterization of HRP dendrozyme was performed by means of circular dichroism and fluorescence spectroscopy to evaluate the secondary and tertiary structure changes after enzyme modification. These investigations revealed that protein conformation had small changes (in secondary and tertiary structures) after bio-conjugation. We also reported here that dendritic modification did not significantly affect the kinetic parameters of free HRP. The stabilization of HRP with dendron macromolecules as single enzyme nanoparticles resulted in improvement of half-life over 70 days storage at 4 °C as well as its tolerance under different elevated temperatures up to 80 °C and in the presence of organic solvents for 15 min. These significant results promise extensive applications of HRP particularly in harsh environmental conditions.     

Applications of stem cells in orthodontics and dentofacial orthopedics:Current trends and future perspectives

A simple overview of daily orthodontic practice involves use of brackets, wires and elastomeric modules. However, investigating the underlying effect of orthodontic forces shows various molecular and cellular changes. Also, orthodontics is in close relation with dentofacial orthopedics which involves bone regeneration. In this review current and future applications of stem cells(SCs) in orthodontics and dentofacial orthopedics have been discussed. For craniofacial anomalies, SCs have been applied to regenerate hard tissue(such as treatment of alveolar cleft) and soft tissue(such as treatment of hemifacial macrosomia). Several attempts have been done to reconstruct impaired temporomandibular joint. Also, SCs with or without bone scaffolds and growth factors have been used to regenerate bone following distraction osteogenesis of mandibular bone or maxillary expansion. Current evidence shows that SCs also have potential to be used to regenerate infrabony alveolar defects and move the teeth into regenerated areas. Future application of SCs in orthodontics could involve accelerating tooth movement, regenerating resorbed roots and expanding tooth movement limitations. However, evidence supporting these roles is weak and further studies are required to evaluate the possibility of these ideas.     

Genetic structure of gall oak (<Emphasis Type="Italic">Quercus infectoria</Emphasis>) characterized by nuclear and chloroplast SSR markers

Quercus infectoria, commonly known as gall oak, is a small shrub found in Iran. Unfortunately, it is subjected to genetic erosion, and so, its conservation and evaluation are desirable. Thus, in the current research, 16 microsatellite primer pairs (seven nuclear simple sequence repeats (nSSRs) and nine chloroplast simple sequence repeats (cpSSRs)) were used in an attempt to assess the genetic diversity of 121 individuals of Q. infectoria belonging to 11 populations from three provinces in northern Zagros forests of Iran. In total, 69 alleles of nSSR and 18 alleles of cpSSR were detected among the individuals. The results of the overall analysis of molecular variance based on nSSRs indicated that 89.00% of the variation was due to differences within populations and 11.00% occurred among populations, while according to cpSSRs, 94.00% of the variation resided among populations, and only 6.00% could be attributed to variation within populations. A higher genetic differentiation of Q. infectoria populations was found according to cpSSR data in comparison to nSSR data. Cophenetic correlation coefficient values were statistically insignificant between nSSR and cpSSR data. The unweighted pair group method with arithmetic mean and Bayesian cluster analyses grouped the studied individuals into two main clusters based on both nSSR and cpSSR data. nSSR data could not completely clustered individuals next each other according to their geographical collection area. Information detailed by nSSR loci revealed that north-Zagros gall oak preserves average levels of genetic diversity at the species level, high level of within-population genetic diversity, and moderate level of genetic variation among populations. The present results provide valuable data for in situ or ex situ conservation and utilization of the studied germplasm.     

A new species of Xiphinema americanum group (Nematoda: Longidoridae) from Iran,with additional data on three known species

Systematic Parasitology - One new and three known species of the genus Xiphinema from the rhizosphere of fruit trees and rose shrubs in East Azarbaijan province, Iran, are presented based on the...     

Effect of timing of application Pseudomonas fluorescens in suppression Sclerotinia sclerotiorum, the causal agent of white mold in canola

Biological control of sclerotinia disease, as an important alternative to chemical control, has received considerable attention due to the lack of resistant varieties in most crop, and increasing concerns over fungicide resistance in population of Sclerotinia sclerotiorum, and fungicide residues in the environment. One biocontrol agent, Pseudomonas fluorescens PB-3, has been showed the antagonistic relationship between itself and S. sclerotiorum was investigated in this study. A petal infection technique was used to detect efficacy of timing of application strain PB-3 in the suppression of S. sclerotiorum on canola. Significant difference in disease severity (p<0.05) were found with respect to timing of ascospore applications in the control treatments (ascospores only). The superior competitive ability strain PB-3 was demonstrated by its complete suppression of disease severity when applied as a co-inoculation treatment or prior to ascospores inoculation. Analysis of effect of applying strain PB-3 after ascospores was indicated that treatment in which strain PB-3 was added to petals 48 or 24 h after ascospores, or when there were no bacteria present at all, had higher rates of disease progression. It would be appear that bacteria are able to significantly inhibit disease when applied before or even at the same time as ascospores. In a practical sense, this could mean that a field application of antagonist could be concurrent with infection by the pathogen.     

A randomized clinical trial on the antitumoral effects of low molecular weight heparin in the treatment of esophageal cancer

The current treatment approaches for esophageal cancer are associated with poor survival, and there are ongoing efforts to find new and more effective therapeutic strategies. There are several reports on the antitumoral effects of low-molecular-weight heparins (LMWHs). We have assessed the possible survival benefit of LMWHs in esophageal malignancies. This was a randomized, single-blind, multicenter, Phase II clinical trial on nonmetastatic esophageal cancer candidate for neoadjuvant chemoradiotherapy. Patients were randomly assigned to the chemoradiotherapy-only arm or chemoradiotherapy plus enoxaparin arm using 1:1 allocation. Radiotherapy was delivered in 1.8-Gy daily fractions to a dose of 50.4 Gy in both groups. Paclitaxel 50 mg/m² and carboplatin (AUC 2) were administered weekly, concurrent with radiotherapy. In the intervention group, patients received enoxaparin (40 mg) and chemoradiation daily. 4–6 weeks after treatment, all patients underwent esophagectomy. After a median follow up of 7 months, estimated 1 year disease-free survival (DFS) in the intervention group was 78.9% and was 70% in the control groups ( p = 0.5). Toxicity from the experimental treatment was minimal, and there were no treatment-related deaths. A pathologically complete response in intervention and control group was 64.8% and 62.5%, respectively ( p = 0.9). There was a nonsignificant trend toward improved survival by the addition of enoxaparin to the concurrent chemoradiotherapy regimen. However, 1 y DFS of both groups were high as expected. A longer follow-up and a larger sample size are required.     

Gene Delivery to Tobacco Root Cells with Single-Walled Carbon Nanotubes and Cell-Penetrating Fusogenic Peptides

Development of efficient, easy, and safe gene delivery methods is of great interest in the field of plant biotechnology. Considering the limitations of the usual transfection methods (such as transgene size and plant type), several new techniques have been tested for replacement. The success of some biological and synthetic nanostructures such as cell-penetrating peptides and carbon nanotubes in transferring macromolecules (proteins and nucleic acids) into mammalian cells provoked us to assess the ability of an engineered chimeric peptide and also arginine functionalized single-walled carbon nanotube in gene delivery to intact tobacco (Nicotiana tabacum var. Virginia) root cells. It was suggested that the engineered peptide with its special cationic and hydrophobic domains and the arginine functionalized single-walled carbon nanotube due to its nano-cylindrical shape can pass plant cell barriers while plasmid DNA (which codes green fluorescent protein) has been condensed on them. The success of gene delivery to tobacco root cells was confirmed by fluorescence microscopy and western blotting analysis.