Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
Asian Pacific Journal of Cancer Prevention
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v.16
no.14
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pp.5635-5641
/
2015
Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.
Brevibacillus brevis GZDF3 is a gram-positive, plant growth-promoting rhizosphere bacterium (PGPR) isolated from the rhizosphere soil of Pinellia ternata (an important herb in traditional Chinese medicine). The GZDF3 strain produces certain active compounds, such as siderophores, which are the final metabolite products of non-ribosomal peptide synthetase (NRPS) and independent non-ribosomal peptide synthetase (NIS) activity. With the present study, we attempted to investigate the siderophore production characteristics and conditions of Bacillus sp. GZDF3. The antibacterial activity of the siderophores on pathogenic fungi was also investigated. Optimal conditions for the synthesis of siderophores were determined by single factor method, using sucrose 15 g/l, asparagine 2 g/l, 32℃, and 48 h. The optimized sucrose asparagine medium significantly increased the production of siderophores, from 27.09% to 54.99%. Moreover, the effects of different kinds of metal ions on siderophore production were explored here. We found that Fe3+ and Cu2+ significantly inhibited the synthesis of siderophores. The preliminary separation and purification of siderophores by immobilized-metal affinity chromatography (IMAC) provides strong antibacterial activity against Candida albicans. The synergistic effect of siderophores and amphotericin B was also demonstrated. Our results have shown that the GZDF3 strain could produce a large amount of siderophores with strong antagonistic activity, which is helpful in the development of new biological control agents.
308 isolates of Ampelomyces sp. were isolated from powdery mildew fungi of 73 plant species in Korea for selection of biocontrol agents. An isolate 94013 isolated from powdery mildew fungus of red bean was selected as an effective biological control agent against cucumber powdery mildew in greenhouse. The morphological characteristics of the isolate is as follows. Pycnidia were sub-globose or elongated to pyriform, pale to dart brown, $52.5{\sim}82.5\;{\times}\;35.0{\sim}47.5\;(ave.\;62.5{\times}40.5){\mu}m$, and conidia were guttulate, straight cylindrical to fusiform, pale brown, $5.0{\sim}8.0\;{\times}\;2.5{\sim}4.3\;(ave.\;6.0{\times}3.0){\mu}m$. The isolate 94013 was identified as Ampelomyces quisqualis by morphological characteristics and rDNA ITS sequencing. The isolate A. quisqualis 94013 was different from that of the commercial product $AQ10^R$ in the rDNA ITS sequence.
Park, Cheol;Jin, Cheng-Yun;Choi, Byung-Tae;Lee, Won-Ho;Choi, Yung-Hyun
Journal of Life Science
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v.18
no.3
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pp.336-343
/
2008
Histone deacetylases (HDACs) inhibitors have emerged as the accessory therapeutic agents for various human cancers, since they can block the activity of specific HDACs, restore the expression of some tumor suppressor genes and induce cell differentiation, cell cycle arrest and apoptosis in vitro and in vivo. In the present study, we investigated that the effect of trichostatin A (TSA), an HDAC inhibitor, on the cell growth and apoptosis, and its effect on the nuclear factor-kappaB $(NF-{\kappa}B)$ activity in 267B1 human prostate epithelial cells. Exposure of 267B1 cells to TSA resulted in growth inhibition and apoptosis induction in and dose-dependent manners as measured by fluorescence microscopy, agarose gel electrophoresis and flow cytometry analysis. TSA treatment inhibited the levels of IAP family members such as c-IAP-1 and c-IAP-2 and induced the proteolytic activation of caspase-3, -8 and -9, which were associated with concomitant degradation of poly (ADP-ribose)-polymerase, ${\beta}-catenin$ and laminin B proteins. The increase in apoptosis by TSA was connected with the translocation of $NF-{\kappa}B$ from cytosol to nucleus, increase of the DNA binding as well as promoter activity of $NF-{\kappa}B$, and degradation of cytosolic inhibitor of KappaB $(I{\kappa}B)-{\alpha}$ protein. We therefore concluded that TSA demonstrated anti-proliferative and apoptosis-inducing effects on 267B1 cells in vitro, and that the activation of caspases and $NF-{\kappa}B$ may play important roles in its mechanism of action. Although further studies are needed, these findings provided important insights into the possible molecular mechanisms of the anti-cancer activity of TSA.
Background: Endophytic fungi play an important role in balancing the ecosystem and boosting host growth. In the present study, we investigated the endophytic fungal diversity of healthy Panax notoginseng and evaluated its potential antimicrobial activity against five major phytopathogens causing root-rot of P. notoginseng. Methods: A culture-dependent technique, combining morphological and molecular methods, was used to analyze endophytic fungal diversity. A double-layer agar technique was used to challenge the phytopathogens of P. notoginseng. Results: A total of 89 fungi were obtained from the roots, stems, leaves, and seeds of P. notoginseng, and 41 isolates representing different morphotypes were selected for taxonomic characterization. The fungal isolates belonged to Ascomycota (96.6%) and Zygomycota (3.4%). All isolates were classified to 23 genera and an unknown taxon belonging to Sordariomycetes. The number of isolates obtained from different tissues ranged from 12 to 42 for leaves and roots, respectively. The selected endophytic fungal isolates were challenged by the root-rot pathogens Alternaria panax, Fusarium oxysporum, Fusarium solani, Phoma herbarum, and Mycocentrospora acerina. Twenty-six of the 41 isolates (63.4%) exhibited activity against at least one of the pathogens tested. Conclusion: Our results suggested that P. notoginseng harbors diversified endophytic fungi that would provide a basis for the identification of new bioactive compounds, and for effective biocontrol of notoginseng root rot.
The experiments were carried out to investigate the biocontrol potential of Lysobacter capsici YS1215 on root-knot nematode and to characterize its lytic enzyme activities. L. capsici YS1215 showed chitinase and gelatinase activities on the medium containing 0.5% chitin or 0.5% gelatin as substrates. Cell growth of L. capsici YS1215 was highest at 6 days, and the highest activities of chitinase (4.0 unit $ml^{-1}$) and gelatinase (7.43 unit $ml^{-1}$) were observed on 3 and 5 days after incubation, respectively. To investigate the effect of L. capsici YS1215 on tomato growth and nematode infection, the plants in pot trial were treated with bacterial culture (BC), half of bacterial culture (HBC), only bacterial medium (BM), tap water (TW) and commercial nematicide (CN). HBC treatd plants showed the higher shoot fresh weight and dry weight on $5^{th}$week after incubation while BM, HBC and BC had consistently higher values than TW at $9^{th}$ week. HBC appeared to be the highest shoot fresh length at $9^{th}$ week. Both CN and BC showed lower number of egg mass, root gall, and population of juveniles in soil compared to BC, HBC, BM and TW. These results suggest that L. capsici YS1215 with its strong ability of lytic enzyme production can be one of the most significant candidates for biocontrol agents against root-knot nematodes.
Park, Kwon-moo;Li, Long-hua;Han, Seong-kyu;Ryu, Pan-dong
Korean Journal of Veterinary Research
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v.39
no.1
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pp.77-84
/
1999
Anesthetic agents are useful in inducing the anesthesia for surgical operations and various biological experiments, but they can disturb the body homeostasis and cause the stress in animals. Much efforts have been directed on reducing such side effects of anesthesia. In this work, we measured the serum ACTH, corticosterone and glucose concentration in rabbits to compare the degree of stress induced by two commonly-used anesthetics, ketamine, xylazine, and the combination of xylazine and ketamine. 1. The anesthesia was induced in about 10 min in the rabbits treated with xyalzine, ketamine and xylazine-ketamine. The duration of complete loss of righting reflex were 12, 13 and 115 min in the groups treated with xylazine, ketamine and xylazine-ketamine, respectively. 2. Serum ACTH concentrations in all treatment groups were higher than those in control group. At 30 min after the administration of the drugs, serum ACTH levels in ketamine-treated group were significantly higher than those in control, xylazine- and xylazine-ketamine-treated groups. However, at 1, 2, 5 and 9 hours after the drug administration, serum ACTH levels in xylazine-treated-group were higher than those in control. 3. Serum corticosterone levels in xylazine- and xylazine-ketamine-treated groups were lower than those in control or ketamine-treated groups at 0.5 and 1 hour after the administration. However, at 5 and 9 hours after the administration, serum corticosterone levels in xylazine- and xylazine-ketamine-treated groups were significantly higher than those in ketamine-treated group or control. 4. Serum glucose levels transiently increased to 3 times of the pre-injection levels at 0.5 and 1 hours after the administration in xylazine or xylazine-ketamine-treated groin, but were not changed in control and ketamine-treated group. These results indicate that xylazine-induced stress lasts longer than ketamine-induced, suggesting that the difference in stress-related hormone levels during anesthesia could be due to the differences in modes of actions of individual drugs used and the depth of anesthesia.
Lysobacter antibioticus HS124 was isolated from rhizosphere soil in previous experiments, which produced lytic enzymes such as chitinase, gelatinase, lipase and protease. In addition, HS124 released an antibiotic compound, 4-hydroxyphenylacetic acid (4-HPAA). When larvae of P. xylostella was treated with HS124 culture broth, its body was destroyed, and degraded with the increase of incubation time, yielding glycine which was detected from HS124 culture broth. When 4-HPAA produced from HS124 was sprayed, larvae mortality increased with increasing concentration of 4-HPAA. When HS124 culture supplemented with Tween 80 was sprayed, its insecticidal activity against larvae was approximately 1.4 times higher compared to the culture without Tween 80. Insecticide (IS), HS124 culture broth (HS124), Magic-pi (MP) and HS124 culture broth+Magic-pi (HS124+MP) were each treated against larvae of P. xylostella to investigate their insecticidal effect where sterile diluted water (SDW) was used as a control. The highest mortality of larvae was found in HS124+MP, followed by IS, MP, HS124 and SDW respectively. Mortality of larvae in HS124 was 31% higher than that in SDW, but 41% lower than that in HS124+MP, meaning that both enzymes and antibiotics produced from HS124 may synergistically act as active agents with plant extract containing neem oil and turmeric in HS124+MP treatment. These results suggested that L. antibioticus HS124 together with plant extract can be one of candidates for biocontrol agents against Plutella xylostella.
Kim, S.I.;Shim, J.O.;Shin, H.S.;Choi, H.J.;Lee, M.W.
The Korean Journal of Mycology
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v.20
no.4
/
pp.337-346
/
1992
Trichoderma spp. are an effective control agent for damping-off or other plant diseases. The interaction between. T. hamatum and Rhizoctonia solani on the rhizosphere or surface soil were examined to assess the possible roles of antibiosis or competition in the mechanisms of biological control agents as a basic research. In a proportional comparison, total bacteria, fungi, actinomycetes and Trichoderma spp were 65%, 8.8%, 25.9% and 0.28% respectively in their distribution in the soil. Among Trichoderma spp isolated, the 5 species of Trichoderma spp were indentified as T. koninggii, T. pseudokoninggii, T. aureoviridi, T. hamatum and T. viride respectively. In a mycoparasitic test, one isolate of T. hamatum strain Tr-5 showed an enzymatic ability to break fungal hyphae into piecies and infected on the R. solani hyphae showing a parasitism. Spore germination of the all isolates of Trichoderma spp showed a 1.7-7.3% of germination in natural soil conditions, but the percentage was high in sterile soil indicating all the natural soil were fungistatic on conidia of Trichoderma spp. In rhizosphere competent assay in pea plant, the antagonistic T. hamatum, T. viride, T. koninggii, T. pseudokoninggii showed a colonizing upper soil depth in rhizosphere around 1-3 cm in root zone, but the colonizing ability was much reduced along the deeper the soil depth. Propagule density was decreased in deeper the soil layer. Disease development rate treated alone with plant pathogens, Fusarium solani, Rhizoctonia solani, Cylindrocarpon destructans increased, but disease incidence rate reduced in treatment with combinations with antagonistic T. hamatum strain Tr-5.
This study was designed to evaluate the anti-diabetic effects of Buan mulberries by using an insulin-dependent diabetes mellitus animal model. Several studies have shown that mulberries have metabolism-improving, antioxidant, and lipid-lowering properties in rats with streptozotocin (STZ)-induced diabetes. In this study, Sprague-Dawley male rats were randomly assigned to 1 normal control group and 5 STZ-induced diabetes groups: rats that had STZ-induced diabetes and did not receive any agents (diabetic group; negative control), rats that had STZ-induced diabetes and received insulin (insulin group; positive control), rats that had STZ-induced diabetes and received 0.5% mulberry extract (0.5% mulberry group), rats that had STZ-induced diabetes and received 1.0% mulberry extract (1.0% mulberry group), and rats that had STZ-induced diabetes and received 2.0% mulberry extract (2.0% mulberry group). Mulberry extracts were administered to the diabetic animals for 4 weeks. The rats that received mulberry extracts showed lower body weights and insulin levels, as well as higher kidney weights, blood glucose levels, urine quantities, and water intake in comparison with the normal controls. Further, the insulin concentrations in the mulberry-fed animals were higher than those in the diabetic group, and the kidney weights, blood glucose levels, urine quantities, and water intake in the mulberry-fed animals were lower than the corresponding values in the diabetic controls. These results suggest that mulberry may be an effective functional food to prevent diabetes-related complications.
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