Genome-Wide Distribution and Organization of Microsatellites in Plants: An Insight into Marker Development in Brachypodium

Plant genomes are complex and contain large amounts of repetitive DNA including microsatellites that are distributed across entire genomes. Whole genome sequences of several monocot and dicot plants that are available in the public domain provide an opportunity to study the origin, distribution and evolution of microsatellites, and also facilitate the development of new molecular markers. In the present investigation, a genome-wide analysis of microsatellite distribution in monocots (Brachypodium, sorghum and rice) and dicots (Arabidopsis, Medicago and Populus) was performed. A total of 797,863 simple sequence repeats (SSRs) were identified in the whole genome sequences of six plant species. Characterization of these SSRs revealed that mono-nucleotide repeats were the most abundant repeats, and that the frequency of repeats decreased with increase in motif length both in monocots and dicots. However, the frequency of SSRs was higher in dicots than in monocots both for nuclear and chloroplast genomes. Interestingly, GC-rich repeats were the dominant repeats only in monocots, with the majority of them being present in the coding region. These coding GC-rich repeats were found to be involved in different biological processes, predominantly binding activities. In addition, a set of 22,879 SSR markers that were validated by e-PCR were developed and mapped on different chromosomes in Brachypodium for the first time, with a frequency of 101 SSR markers per Mb. Experimental validation of 55 markers showed successful amplification of 80% SSR markers in 16 Brachypodium accessions. An online database ‘BraMi’ (Brachypodium microsatellite markers) of these genome-wide SSR markers was developed and made available in the public domain. The observed differential patterns of SSR marker distribution would be useful for studying microsatellite evolution in a monocot–dicot system. SSR markers developed in this study would be helpful for genomic studies in Brachypodium and related grass species, especially for the map based cloning of the candidate gene(s)

Combined Tumor Cell-Based Vaccination and Interleukin-12 Gene Therapy Polarizes the Tumor Microenvironment in Mice

Tumor progression depends on tumor milieu, which influences neovasculature formation and immunosuppression. Combining immunotherapy with antiangiogenic/antivascular therapy might be an effective therapeutic approach. The aim of our study was to elaborate an anticancer therapeutic strategy based on the induction of immune response which leads to polarization of tumor milieu. To achieve this, we developed a tumor cell-based vaccine. CAMEL peptide was used as a B16-F10 cell death-inducing agent. The lysates were used as a vaccine to immunize mice bearing B16-F10 melanoma tumors. To further improve the therapeutic effect of the vaccine, we combined it with interleukin (IL)-12 gene therapy. IL-12, a cytokine with antiangiogenic properties, activates nonspecific and specific immune responses. We observed that combined therapy is significantly more effective (as compared with monotherapies) in inhibiting tumor growth. Furthermore, the tested combination polarizes the tumor microenvironment, which results in a switch from a proangiogenic/immunosuppressive to an antiangiogenic/immunostimulatory one. The switch manifests itself as a decreased number of tumor blood vessels, increased levels of tumor-infiltrating CD4+, CD8+ and NK cells, as well as lower level of suppressor lymphocytes (Treg). Our results suggest that polarizing tumor milieu by such combined therapy does inhibit tumor growth and seems to be a promising therapeutic strategy

Anti-angiogenic alternatives to VEGF blockade

Angiogenesis is a major requirement for tumour formation and development. Anti-angiogenic treatments aim to starve the tumour of nutrients and oxygen and also guard against metastasis. The main anti-angiogenic agents to date have focused on blocking the pro-angiogenic vascular endothelial growth factors (VEGFs). While this approach has seen some success and has provided a proof of principle that such anti-angiogenic agents can be used as treatment, the overall outcome of VEGF blockade has been somewhat disappointing. There is a current need for new strategies in inhibiting tumour angiogenesis; this article will review current and historical examples in blocking various membrane receptors and components of the extracellular matrix important in angiogenesis. Targeting these newly discovered pro-angiogenic proteins could provide novel strategies for cancer therapy

<span style="font-size:14.0pt;line-height: 115%;font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; color:black;mso-ansi-language:EN-IN;mso-fareast-language:EN-IN;mso-bidi-language: HI" lang="EN-IN">Soybean (<i>Glycine max </i>L.) Seed Coat Peroxidase Immobilized on Fibrous Aromatic Polyamide: A Strategy for Decreasing Phenols from Industrial Wastewater</span>

1090-1093Peroxidase enzyme extracted from soybean (Glycine max L.) seed coats and immobilized on fibrous aromatic polyamide are used for the detoxification of phenols from industrial wastewater. It is observed that during 5 h treatment with immobilized peroxidase the phenolic concentration is reduced from 0.720 to 0.063 mg/mL

Antioxidant potential of selected vegetables commonly used in diet in Asian subcontinent

306-313In the present study eleven different fruits and leaves of commonly used vegetables in diet in Asian subcontinent have been evaluated for antioxidative constituents and free radical scavenging activities. Fifty per cent ethanolic extracts of Abelmoschus esculentus (Linn.) Moench (fruits), Trigonella foenum-graecum Linn. (leaves), Spinacia oleracea Linn. (leaves), Brassica oleracea Linn. var. capitata and B. oleracea var. botrytis (leaves, inflorescence and young stems), Coriandrum sativum Linn. (seeds and leaves), Capsicum annuum Linn. var. grossum (Willd.) Sendt. (fruits), Cucurbita maxima Duch. (fruits), Cyamopsis tetragonoloba Linn. (fruits), Anethum graveolens Linn. (fruits and seeds), Solanum melongena Linn. (fruits) were tested for the determination of free radical scavenging potentials and quantification of antioxidant agents such as ascorbic acid and phenolic compounds. Amongst the tested vegetables the sample of B. oleracea var. botrytis has shown (67.2%) the highest 1, 1-Diphenyl-2-picryl hydrazine (DPPH radical) scavenging potential while B. oleracea var. capitata was found to be the most effective (58.4%) inhibitor of lipid peroxidation. The extract of S. oleracea (43.9%) and S. melongena (32.8%) were found to be effective in ferrous ion chelating abilities. The maximum amount (25.60mg/100g) of vitamin-C was found in A. esculentus while the amount of total phenolics was noted maximum (13.30 mg/g) in C. maxima. With some exceptions the activity profiles of all other samples were found to be good to moderate

Potato Phenol Oxidases Immobilized s-Triazine Containing Dialdehyde: An Efficient Biocatalysts For Bioremediation of Phenols

621-624Polyphenol oxidase (PPO) and peroxidase enzymes extracted from potato and immobilized on s- triazine have been exploited for the detoxification of phenols from industrial effluents. It was found that the phenol content of industrial effluent was reduced from 168 to 38 μg/mL within three and half hours treatment with immobilized phenol oxidases

Reversible inhibition of polyphenol oxidase from apple using L-cysteine

459-463Alternate strategies for prevention of enzymatic browning of fruits and vegetables are required by food industries due to restrictions and several negative health attributes of sulphites, which are effective inhibitors of enzymatic browning reaction. These reactions result mostly from polyphenol oxidase (PPO). The present study was conducted to evaluate the antioxidant and PPO inhibitory potential of L-cysteine. Studies have revealed that with increase in concentration of L-cysteine there was corresponding decrease in the rate of browning reaction and the activity of PPO. The kinetic study indicates that PPO inhibition by L-cysteine is non-competitive. The results suggest that the L-cysteine is a time bound inhibitor of PPO, since its highest concentration (5 mM) gives protection up to 4 h having no further effect on the rate of browning reaction in apple juice. The assessment of radical-scavenging ability of L-cysteine demonstrated that it has high affinity towards a stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH·)