The 6-Methoxymethyl Derivative Of Pyrrolo-Dc For Selective Fluorometric Detection Of Guanosine-Containing Sequences

The β-cyanoethyl phosphosphoramidite derivatives of 6-methyl- and 6-methoxymethyl-3-(2-deoxy-β-D-ribofuranosyl)-3H-pyrrolo[2,3-d]pyrimidin-2-one have been synthesized. These monomers have been employed for oligodeoxynucleotide synthesis to evaluate their effect on duplex stability and ability to fluorometrically report on hybridization. The structurally conservative 6-methoxymethyl-substitution results in a pyrrolocytidine that is stabilizing toward hybrid formation (Δ Tm = +1.3 °C) whereas the known 6-methylpyrrolocytidine is destabilizing (Δ Tm = ?4.7 °C), in the sequence examined. The 6-methoxymethylpyrrolocytidine retains excellent mismatch discrimination and its fluorescence is selectively quenched when hybridized to a match oligodeoxynucleotide sequence. The quenching of fluorescence for an internal position is approximately three-fold, whereas a terminal position (5′-end or 3′-end) experienced approximately two-fold decrease in the fluorescence intensity.     

Construction of an expression plasmid (vector) encoding Brucella melitensis outer membrane protein,a candidate for DNA vaccine

Background:

DNA vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. Bacteria of the genus Brucella are facultative intracellular pathogens for which new and efficient vaccines are needed.

Methods:

To evaluate the use of a DNA immunization strategy for protection against brucellosis, a plasmid containing the DNA encoding the Brucella melitensis (B. melitensis) 31 kDa outer membrane protein, as a potent immunogenic target, was constructed.

Results:

The constructed plasmid, pcDNA3.1+omp31, was injected intramuscularly into mice and the expression of omp31 RNA was assessed by RT-PCR. The integrity of the pcDNA3.1+omp31 construct was confirmed with restriction analysis and sequencing. Omp31 mRNA expression was verified by RT-PCR.

Conclusion:

Our results indicate that the pcDNA3.1+omp31 eukaryotic expression vector expresses omp31 mRNA and could be useful as a vaccine candidate.Key Words: Brucella melitensis, omp31, DNA Vaccine, pcDNA3.1     

Modelling cellular signal communication mediated by phosphorylation dependent interaction with 14-3-3 proteins

The 14-3-3 proteins are important effectors of Ser/Thr phosphorylation in eukaryotic cells. Using mathematical modelling we investigated the roles of these proteins as effectors in signalling pathways that involve multi-phosphorylation events. We defined optimal conditions for positive and negative cross-talk. Particularly, synergistic signal interaction was evident at very different sets of binding affinities and phosphorylation kinetics. We identified three classes of 14-3-3 targets that all have two binding sites, but displayed synergistic interaction between converging signalling pathways for different ranges of parameter values. Consequently, these protein targets will respond differently to interventions that affect 14-3-3 binding affinities or phosphorylation kinetics.     

Health Risks from Heavy Metals via Consumption of Cereals and Vegetables in Isfahan Province,Iran

The present study was conducted to assess the risks to human health from exposure to arsenic, lead, nickel, zinc, and copper through consumption of wheat, rice, and some vegetables in Isfahan Province, central Iran, using the total non-carcinogenic hazard quotient and cancer risk assessment estimates. Risk contribution from Ni is minimal compared with other metals and it was less than the U.S. Environmental Protection Agency guidelines. The results showed that total non-carcinogenic hazard of As and Pb, were greater than 1, and total cancer risk of As slightly greater than 1 × 10^?4 for all age groups. Therefore, the inhabitants who reside in Isfahan Province may experience the adverse health risks via consumption of cereals and vegetables.     

Decoration of electrical conductive polyurethane-polyaniline/polyvinyl alcohol matrixes with mussel-inspired polydopamine for bone tissue engineering

Electrospinning is a versatile technology for the fabrication of nanofibrous matrixes to regenerate defects. This study aims to develop a functionalized and electroconductive polymeric matrix to improve rat bone marrow mesenchymal stem cell adhesion, proliferation, and differentiation. Herein, the influence of the chemical composition of the substrate on homogeneous modification of the surface with mussel-inspired polydopamine (PDA) is focused. Accordingly, the deposition of PDA on the surface was proved by Fourier transform infrared spectroscopy. Morphologies of the scaffolds demonstrated homogeneous decoration of the polyvinyl alcohol (PVA)/polyurethane (PU)-polyaniline (PANI) matrixes with PDA, while a lower density of mussel-inspired polymer was observed in bare PU-PANI constructs. Although uniform and dense precipitation of PDA reduced conductivity of scaffolds 1.2 times compared with the samples with a low density of the PDA, 1.1 and 1.2 times enhancement in tensile strength and Young's modulus, respectively, were the strength of the applied process, especially in bone tissue engineering area. Contact angle measurements demonstrated about two times reduction in measured values, which shows improvement in hydrophilicity of PDA-modified PVA/PU-PANI fibers compared with PDA-coated PU-PANI ones. Swelling ratio and mass loss ratio calculations revealed enhancement in measured values as a function of homogeneous and dense coating, which arise from hydrophilicity of the polymeric substrate. The bioactivity test indicated that a dense layer of PDA strongly supports formations of hydroxyapatite-like crystals. Moreover, homogeneous decoration of conductive matrixes with PDA showed suitable cell viability, adhesion, and spreading while cell-scaffolds interactions improved under electrical stimulation. Higher expression of alkaline phosphatase and secretion of Collagen I under the electrical field proved the applicability of modified electroconductive scaffolds for further preclinical and clinical studies to introduce as a reconstructive bone substitute.     

Distribution of indole-3-acetic acid in Petunia hybrida shoot tip cuttings and relationship between auxin transport, carbohydrate metabolism and adventitious root formation

To determine the contribution of polar auxin transport (PAT) to auxin accumulation and to adventitious root (AR) formation in the stem base of Petunia hybrida shoot tip cuttings, the level of indole-3-acetic acid (IAA) was monitored in non-treated cuttings and cuttings treated with the auxin transport blocker naphthylphthalamic acid (NPA) and was complemented with precise anatomical studies. The temporal course of carbohydrates, amino acids and activities of controlling enzymes was also investigated. Analysis of initial spatial IAA distribution in the cuttings revealed that approximately 40 and 10 % of the total IAA pool was present in the leaves and the stem base as rooting zone, respectively. A negative correlation existed between leaf size and IAA concentration. After excision of cuttings, IAA showed an early increase in the stem base with two peaks at 2 and 24 h post excision and, thereafter, a decline to low levels. This was mirrored by the expression pattern of the auxin-responsive GH3 gene. NPA treatment completely suppressed the 24-h peak of IAA and severely inhibited root formation. It also reduced activities of cell wall and vacuolar invertases in the early phase of AR formation and inhibited the rise of activities of glucose-6-phosphate dehydrogenase and phosphofructokinase during later stages. We propose a model in which spontaneous AR formation in Petunia cuttings is dependent on PAT and on the resulting 24-h peak of IAA in the rooting zone, where it induces early cellular events and also stimulates sink establishment. Subsequent root development stimulates glycolysis and the pentose phosphate pathway.     

Inhibitory effect of essential oils of Eucalyptus sp. and Zataria multiflora on fungal growth of Macrophomina phaseolina

Macrophomina phaseolina is a causal agent for charcoal rot and cause economic damage in plants. In recent years, the great tendency for the use of biological substances to control of pests, weeds and the pathogens has been created. In this study, antifungal effects of concentrations 30, 60 and 120?μl/100?ml of essential oil of Eucalyptus sp. and Zataria multiflora were evaluated on the growth of M. phaseolina in vitro condition. The experiment was conducted on phactorial basis of completely randomised design with four replications. According to the results, a significant difference between inhibitory percentage of essential oils and different concentrations of essential oils on fungal colony growth has been observed (p?≤?0.01). Highest inhibitory percentage was related to the concentration of 120?μl/100?ml of Z. multiflora with 58.16% inhibition.     

A comparison of the wild food plant use knowledge of ethnic minorities in Naban River Watershed National Nature Reserve, Yunnan, SW China

ABSTRACT: BACKGROUND: Wild food plants (WFPs) contribute to the nutrition, economy and even cultural identity of people in many parts of the world. Different factors determine the preference and use of WFPs such as abundance, availability, cultural preference, economic conditions, shortage periods or unsecure food production systems. Understanding these factors and knowing the patterns of selection, use and cultural significance and value of wild food plants for local communities is helpful in setting priorities for conservation and/or domestication of these plants. Thus in this study knowledge of wild food plant use among four groups namely Dai, Lahu, Hani and Mountain Han in Naban River Watershed National Nature Reserve ((NRWNNR), Xishuangbanna were documented and analyzed to find the similarity and difference among their plant use. METHODS: Data on wild food plant use was collected through freelisting and semi-structured interviews and participatory field collection and direct observation. Botanical plant sample specimens were collected, prepared, dried and identified. RESULTS: A total of 173 species and subspecies from 64 families and one species of lichen (Ramalina sp.) are used as WFP. There were differences on the saliency of wild food plant species among four ethnic groups. Consensus analysis revealed that knowledge of wild food plant use for each ethnic group differs from others with some variation in each group. Among informant attributes only age was related with the knowledge if wild food plant use, whereas no significant relationship was found between gender and age*gender and informants knowledge of wild food plant use. CONCLUSION: Wild food plants are still used extensively by local people in the NRWNNR, some of them on a daily base. This diversity of wild food plants provide important source of nutrients for the local communities which much of their caloric intake comes from one or few crops. The results also show the role of ethnicity on the preference and use of wild food plants. There is a big potential for harvesting, participatory domestication and marketing of WFPs especially in the tourism sector in the area.     

Compensatory growth in juvenile roach Rutilus caspicus: effect of starvation and re‐feeding on growth and digestive surface area

The aim of this study was to investigate compensatory growth in juvenile Rutilus caspicus during starvation and re‐feeding periods. The results confirmed the existence of compensatory growth in R. caspicus which depended on the duration of food deprivation. Complete compensatory growth occurred in the fish that were food deprived for at least 3 weeks. Starvation and re‐feeding had no significant effect on the digestive somatic index and intestinal surface areas in the fish that were food deprived for 1 week, while they showed a significant decrease and increase, during starvation and re‐feeding in the fish that were food deprived for 2 and 3 weeks. This knowledge may have application in aquaculture, as appropriate exploitation of compensatory growth can give increased growth rate and feeding efficiency.