Antimicrobial activity of sea Cucumber (Stichopus variegatus) body wall extract in Chabahar Bay, Oman Sea

Background: Seas and oceans have been recognized as a rich source of metabolites and bioactive compounds with biodiversity and drug activity. These compounds have accumulated in various organisms. Bioactive compounds can be extracted from various animal groups, such as coral, crabs, moss animals, Echinodermata, case-bearers, fishes, and sponges. Objectives: Antimicrobial extracts of the sea cucumber body wall (Stichopus variegatus), collected from Chabahar Bay, were tested for four bacterial, fungal, and yeast species. Methods: The disk diffusion method was used in this study. The McFarland 0.5 standard was used in the preparation of suspensions of the mentioned microorganisms under a hood. Next, the species were cultured with a swab on a Mueller-Hinton agar for bacteria and on a Sabouraud dextrose agar for fungi. Subsequently, the disks, with appropriate concentrations, were placed on microbiological growth media. Results: The results of the study showed that the best effect belonged to the aqueous methanol extract of the sea cucumber body wall, with an 8 mg/mL concentration on E. coli, with an inhibition zone diameter of 12.26 mm. The lowest inhibition zone diameter belonged to the methanol extract of white strands, with a 2 mg/mL concentration on C. albicans and an inhibition zone diameter of 1.16 mm. The n-hexane and chloroform extracts had no effect. Conclusions: Comparison of these tests indicates that sea cucumbers have an innate immunity system, which can be considered a potential source for discovering antimicrobial peptides

Antibacterial and antifungal activity of Holothuria leucospilota isolated from Persian Gulf and Oman Sea

Background: Emergence of antimicrobial resistance toward a number of conventional antibiotics has triggered the search for antimicrobial agents from a variety of sources including the marine environment. Objectives: The aim of this study was to evaluate the antimicrobial potential of Holothuria leucospilota from Qeshm and Kharg Islands against some selected bacteria and fungi. Materials and Methods: In this investigation, sea cucumbers from two coastal cities of Persian Gulf were collected in March and May 2011 and identified by the scale method according to the food and agriculture organization of the United Nations. Antibacterial activity of hydroalcoholic extracts of the body wall, cuvierian organs and coelomic fluid, methanol, chloroform, and n-hexane extracts of the body wall were evaluated by the spot test. In addition, their antifungal activity was assessed by the broth dilution method. Results: The displayed effect was microbiostatic at concentrations of 1000 and 2000 µg/mL rather than microbicidal. The highest activity of hydroalcoholic extracts was exhibited by body wall, cuvierian organs and coelomic fluid against Escherichia coli, Salmonella typhi, Staphylococcus aureus and Pseudomonas aeruginosa; Aspergillus niger, A. fumigatus, A. flavus and A. brasilensis. However, none of the methanol, chloroform and n-haxane extracts showed appreciable effects against Shigella dysenteriae, Proteus vulgaris, Bacillus cereus, S. epidermidis and Candida albicans. Moreover, cuvierian organs did not possess any antifungal potential.Conclusions: Our data indicated that water-methanol extracts from the body wall of H. leucospilota possess antibacterial and antifungal activity. However, additional and in-depth studies are required to isolate and identify the active component(s)

Tolerance and Accumulation of Heavy Metals by Descurainia sophia L.

Today, biosphere pollution has accelerated strongly with start of industrial revolution by toxicity of heavy metals. One of existing pollution is soil pollution. Unfortunately, soil pollution by metals is as intensive environmental stress for plant hence for human. Plants, which are able to store heavy metals in their organs, can be used for phytoremediation of polluted soils and utilization of these plants is effective for phytoremediation as a cheap and economic method. In this research, the absorption rate of Cd (II), Ni (II) by Descurainia sophia was considered in hydroponic conditions. Plants were grown in Hoagland media containing different concentrations of Cd (II), Ni (II). An experiment in a completely randomized design with three replications was conducted. Two weeks after treatment of plants the sample were gathered and metal concentration was measured by atomic absorption spectroscopy. Besides, the content of chlorophyll and proline was measured. The results showed the chlorophyll content in high concentrations of the metals (Cd (II), Ni (II)) was decreased in plants that were sign of pigment degradation in presence of heavy metals. Similarly, the proline content in plants was increased under stress which was sign of damage of heavy metal stress on plant and activation of defensive mechanisms in this condition. The effects of toxic concentration of nickel and cadmium on metal accumulation in these plants showed that roots were able to absorb more than shoots, which is sign of elements connection to root cell wall

Effect of Salicylic Acid and Ethephon on Seed Germination and Seedling Growth of Wheat under Salt Stress

Water or soil salinities are the most important factors that reduce the seed germination of plants. Ethephon can break seed dormancy in a variety of plants, such as cereals and speeds up germination. In some plants pretreatment of seeds with salicylic acid has increased the germination percentage. To study effect of salicylic acid and ethephon on seed germination of wheat (Seivand cultivar) under salinity condition a factorial experiment in a completely randomized design with three replications was conducted at the Plant Research Laboratory of Neyshabur Branch of Islamic Azad University in 2011. Four salinity levels (0, 50, 100, 150 mM), three salicylic acid levels (0, 0.5, 1 mM) and four ethephon levels (0, 0.5, 1, 2 mM) were used. The results showed that at salinity condition seed germination rate and percentage, shoot and root length, their dry weight and α-amylase activity decreased and proline content increased. Pretreatment of seeds by salicylic acid increased seed germination percentage, some growth parameters, α-amylase activity and proline content under salinity condition. Moreover, pretreatment of seeds by ethephon decreased some growth parameters and increased proline content but its effect on germination and α-amylase activity were not significant. It seems that Salicylic acid as a plant growth regulator under salinity condition and ethephon convertion to ethylene, activated plant tolerance mechanisms to salinity condition and decrease damaging effect of salinity on seed germination and seedling growth of wheat

Growth inhibition of human breast cancer cells and down-regulation of ODC1 and ADA genes by Nepeta binaloudensis

Nepeta binaloudensis Jamzad, Lamiaceae, is a rare medicinal plant endemic to Iran. In spite of many studies about the chemical constituents and antibacterial effects of this species, no report has been provided about its cytotoxic and anticancer activities. In this study we have evaluated the effects of EtOH 70%, hexane and aqueous extracts of N. binaloudensis on the cell proliferation and n-hexane extract on the expression of adenosine deaminase and ornithine decarboxylase 1 genes in breast cancer cell lines (MCF-7, MDA-MB-231) compared to non-cancer line (MCF-10A). The cell lines were subjected to increasing doses of the extracts ranging from 10 to 320 μg/ml. Cell viability was quantified by MTS assay. Expression of adenosine deaminase and ornithine decarboxylase 1 genes was analyzed by real time PCR. N. binaloudensis inhibited the growth of malignant cells in a time and dose-dependent manner. Among extracts of N. binaloudensis, the hexane extract was found to be more toxic compared to other extracts. Results showed a marked decrease in the expression of ornithine decarboxylase 1 and adenosine deaminase genes in cancer cell lines. At 60 μg/ml concentration of N. binaloudensis hexane extract ornithine decarboxylase 1 and adenosine deaminase mRNA expression were reduced 4.9 fold and 3.5 fold in MCF-7 cell line and 3.6 fold and 2.6 fold in MDA-MB-231 cell line compared to control, respectively. The result of our study highlights the potential influences of N. binaloudensis hexane extract on ornithine decarboxylase 1 and adenosine deaminase genes expression in breast cancer cells and its relation to inhibition of cancer cell growth. Keywords: Adenosine deaminase, Anticancer, Cytotoxicity, Ornithine decarboxylas

EFFECT OF SALICYLIC ACID ON SOME OF MORPHOLOGICAL AND PHYSIOLOGICAL TRAITS OF WHEAT (Triticum aestivum L.) UNDER DIFFERENT LEVELS OF CADMIUM STRESS

EFEITO DE NANO-QUELADO DE NITROGÊNIO E UREIA FERTILIZANTE EM PLANTAS DE TRIGO SOB CONDIÇÕES DE ESTRESSE HÍDRICO Atualmente, o ácido salicílico é usado como um regulador de crescimento para reduzir os efeitos negativos de diferentes níveis de estresse. Esta pesquisa foi conduzida como experimento fatorial em um delineamento de blocos inteiramente casualizados. Plantas de trigo foram plantadas em quatro níveis de cádmio (0, 100, 200 e 300 μmol) em três níveis de ácido salicílico (0, 0,5 e 1,0 mmol) em três repetições. Os resultados mostraram que a altura das raízes, o peso fresco e seco do caule diminuíram significativamente e as enzimas prolina e catalase e superóxido dismutase foram aumentadas devido ao estresse com cádmio. A aplicação foliar de ácido salicílico em condições de não estresse teve um efeito significativo sobre as características e também resultou em seu aumento. A aplicação de duas concentrações de 0,5 e 1,0 mmol de ácido salicílico em condições de estresse resultou na redução dos efeitos do estresse e, consequentemente, na redução das enzimas prolina e catalase e superóxido dismutase e o crescimento foi melhorado. A quantidade de 1,0 mmol de ácido salicílico foi mais efetiva e pode-se concluir que o ácido salicílico tem um efeito moderador e decrescente nos efeitos negativos da toxicidade do cádmio em plantas de trigo.Palavras-chave: trigo (Triricum aestivum L.), ácido salicílico, cádmio.  ABSTRACT: Nowadays, Salicylic acid is used as a growth regulator to reduce the negative effects of different levels of stress. This research was conducted as factorial experiment on a completely randomized block design. Wheat plants were planted in four levels of cadmium (0, 100, 200 and 300 μmol) with three levels of salicylic acid (0, 0.5 and 1 mmol) in three replications. The results showed that root heights, fresh and dry weight of the stem were decreased significantly and leaf proline and catalase and superoxide dismutase enzymes were increased due to cadmium stress. Foliar application of salicylic acid in non-stress conditions had a significant effect on the traits and resulted in their increase, as well. Application of two concentrations of 0.5 and 1 mmol salicylic acid in stress conditions resulted in reduction of the effects of stress and consequently, reduction of proline and catalase and superoxide dismutase enzymes and growth was improved. The amount of 1mmol of salicylic acid was more effective and it can be concluded that salicylic acid has a moderating and decreasing effect on the negative effects of cadmium toxicity in wheat plant.Keywords: wheat (Triricum aestivum L.), salicylic acid, cadmium

Evaluating the efficiency of CHEF and CMV promoter with IRES and Furin/2A linker sequences for monoclonal antibody expression in CHO cells.

In recent years, monoclonal antibodies (mAbs) have been developed as powerful therapeutic and diagnostic agents and Chinese hamster ovary (CHO) cells have emerged as the dominant host for the recombinant expression of these proteins. A critical step in recombinant expression is the utilization of strong promoters, such as the Chinese Hamster Elongation Factor-1α (CHEF-1) promoter. To compare the strengths of CHEF with cytomegalovirus (CMV) promoter for mAb expression in CHO cells, four bicistronic vectors bearing either internal ribosome entry site (IRES) or Furin/2A (F2A) sequences were designed. The efficiency of these promoters was evaluated by measuring level of expressed antibody in stable cell pools. Our results indicated that CHEF promoter-based expression of mAbs was 2.5 fold higher than CMV-based expression in F2A-mediated vectors. However, this difference was less significant in IRES-mediated mAb expressing cells. Studying the stability of the F2A expression system in the course of 18 weeks, we observed that the cells having CHEF promoter maintained their antibody expression at higher level than those transfected with CMV promoter. Further analyses showed that both IRES-mediated vectors, expressed mAbs with correct size, whereas in antibodies expressed via F2A system heterogeneity of light chains were detected due to incomplete furin cleavage. Our findings indicated that the CHEF promoter is a viable alternative to CMV promoter-based expression in F2A-mediated vectors by providing both higher expression and level of stability

Monoclonal antibodies expression improvement in CHO cells by PiggyBac transposition regarding vectors ratios and design.

Establishing stable Chinese Hamster Ovary (CHO) cells producing monoclonal antibodies (mAbs) usually pass through the random integration of vectors to the cell genome, which is sensitive to gene silencing. One approach to overcome this issue is to target a highly transcribed region in the genome. Transposons are useful devices to target active parts of genomes, and PiggyBac (PB) transposon can be considered as a good option. In the present study, three PB transposon donor vectors containing both heavy and light chains were constructed, one contained independent expression cassettes while the others utilized either an Internal Ribosome Entry Site (IRES) or 2A element to express mAb. Conventional cell pools were created by transferring donor vectors into the CHO cells, whereas transposon-based cells were generated by transfecting the cells with donor vectors with a companion of a transposase-encoding helper vector, with 1:2.5 helper/donor vectors ratio. To evaluate the influence of helper/donor vectors ratio on expression, the second transposon-based cell pools were generated with 1:5 helper/donor ratio. Expression levels in the transposon-based cells were two to five -folds more than those created by conventional method except for the IRES-mediated ones, in which the observed difference increased more than 100-fold. The results were dependent on both donor vector design and vectors ratios