• Natural Product Sciences
• /
• v.8 no.1
• /
• pp.1-15
• /
• 2002

An International Cooperative Biodiversity Group (ICBG) program based at the University of Illinois at Chicago initiated its activities in 1998, with the following specific objectives: (a) inventory and conservation of of plants of Cuc Phuong National Park in Vietnam and of medicinal plants of Laos; (b) drug discovery (and development) based on plants of Vietnam and Laos; and (c) economic development of communities participating in the ICBG project both in Vietnam and Laos. Member-institutions and an industrial partner of this ICBG are bound by a Memorandum of Agreement that recognizes property and intellectual property rights, prior informed consent for access to genetic resources and to indigenous knowledge, the sharing of benefits that may arise from the drug discovery effort, and the provision of short-term and long-term benefits to host country institutions and communities. The drug discovery effort is targeted to the search for agents for therapies against malaria (antimalarial assay of plant extracts, using Plasmodium falciparum clones), AIDS (anti-HIV-l activity using HOG.R5 reporter cell line (through transactivation of the green fluorescent protein/GFP gene), cancer (screening of plant extracts in 6 human tumor cell lines - KB, Col-2, LU-l, LNCaP, HUVEC, hTert-RPEl), tuberculosis (screening of extracts in the microplate Alamar Blue assay against Mycobacterium tuberculosis $H_{37}Ra\;and\;H_{37}Rv),$ all performed at UIC, and CNS-related diseases (with special focus on Alzheimer's disease, pain and rheumatoid arthritis, and asthma), peformed at Glaxo Smith Kline (UK). Source plants were selected based on two approaches: biodiversity-based (plants of Cuc Phuong National Park) and ethnobotany-based (medicinal plants of Cuc Phuong National Park in Vietnam and medicinal plants of Laos). At mc, as of July, 2001, active leads had been identified in the anti-HIV, anticancer, antimalarial, and anti- TB assay, after the screening of more than 800 extracts. At least 25 biologically active compounds have been isolated, 13 of which are new with anti-HIV activity, and 3 also new with antimalarial activity. At GSK of 21 plant samples with a history of use to treat CNS-related diseases tested to date, a number showed activity against one or more of the CNS assay targets used, but no new compounds have been isolated. The results of the drug discovery effort to date indicate that tropical plant diversity of Vietnam and Laos unquestionably harbors biologically active chemical entities, which, through further research, may eventually yield candidates for drug development. Although the substantial monetary benefit of the drug discovery process (royalties) is a long way off, the UIC ICBG program provides direct and real-term benefits to host country institutions and communities.

• Korean Journal of Plant Resources
• /
• v.26 no.3
• /
• pp.403-409
• /
• 2013

This study has assessed the anti-oxidative activities and cytotoxic effects of Lithospermum erythrorhizon ethanol extract and measured the effects of tyrosinase inhibition activities with a goal of estimating the usage of the medicinal plant as an ingredient of cosmetics. First, to perform a basic test on the extract, pH and UV-spectrum were measured. According to the measurement, the extract had control functions at pH 5.5, and maximum absorbance occurred at 530nm. In particular, DPPH (1-1-diphenyl-2-picryl-hydrazyl)-inhibiting activity (IC50) and polyphenol content were 149.81 ${\mu}g/mL$ and $51.28{\pm}2.52$ mg/mL respectively. In addition, as extract concentration increased, tyrosinase inhibition activities improved as well. In raw 264.7 cell-based MTT assay, cell survival rates were 98% at 1000 ppm and 153% at 100 ppm. Therefore, it's been confirmed that there is almost no cytotoxin. According to the test results above, it appears that the Lithospermum erythrorhizon ethanol extract would be effective in anti-oxidation and application as a cosmetic ingredient.

• Genomics & Informatics
• /
• v.8 no.4
• /
• pp.201-205
• /
• 2010

The H1 histone family, member N, testis-specific (H1FNT) is exclusively expressed in the testis, and had its possible role for sperm chromatin formation. The purpose of this study is to investigate any genetic association of H1FNT gene with male infertility, especially at the promoter region. We examined the promoter single nucleotide polymorphisms (SNP) of H1FNT gene which is located within transcription factor binding site for its association with male infertility. The statistical analysis showed that the -1129A>T polymorphism was present at a statistically significance in male infertility (p=0.0059 and 0.0349 for hetero and risk type, respectively). The dual-luciferase promoter assay was performed to examine the polymorphic effect of this promoter SNP by the cloning of promoter region (1700bp fragment) into pGL3-basic vector. In our plasmid based reporter system, there is no big difference between wild and risk type. In conclusion, H1FNT -1129A>T promoter SNP is statistically significant with male infertility, especially with subfertile (non-azoospermia) group. Further analysis of its functional polymorphic effect in vivo may provide the biological significance of testis-specific histone with spermatogenesis.

• Journal of Microbiology
• /
• v.44 no.1
• /
• pp.42-49
• /
• 2006

In this study, we describe our newly-developed sensitive two-stage PCR procedure for the detection of 13 common mycoplasmal contaminants (M. arthritidis, M. bovis, M. fermentans, M. genitalium, M. hominis, M. hyorhinis, M. neurolyticum, M. orale, M. pirum, M. pneumoniae, M. pulmonis, M. salivarium, U. urealyticum). For primary amplification, the DNA regions encompassing the 16S and 23S rRNA genes of 13 species were targeted using general mycoplasma primers. The primary PCR products were then subjected to secondary nested PCR, using two different primer pair sets, designed via the multiple alignment of nucleotide sequences obtained from the 13 mycoplasmal species. The nested PCR, which generated DNA fragments of 165-353 bp, was found to be able to detect 1-2 copies of the target DNA, and evidenced no cross-reactivity with the generated DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. The identification of contaminated species was' achieved via the performance of restriction fragment length polymorphism (RFLP) coupled with Sau3AI digestion. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the disagnosis of mycoplasmal contamination in cell culture systems.

• Environmental Mutagens and Carcinogens
• /
• v.22 no.4
• /
• pp.312-318
• /
• 2002

Dioxin represents a group of halogenated aromatic hydrocarbons of which 2,3,7,8-tetrachlorod-ibenzo-p-dioxin (TCDD) is well known for its extremely toxic properties as well as ubiquitous presence in our environment and ecosystems. In order to better assess the carcinogenic mechanism of dioxin, we should utilize the reliable biomarkers that can precisely and correctly reflect multi-stage carcinogenesis. When MCF10A cells were exposed to TCDD (10 nM), expression of both CYP1A1 and CYP1B1 was induced in a time-related manner. The expression as well as activity of ornithine decarboxylase was transiently induced by TCDD treatment. In contrast, the induction of COX-2 that is implicated in carcinogenesis as well as inflammation, was not induced by TCDD. In another study, gap-junctional intercellular communication (GJIC) was attenuated by TCDD treatment as revealed by the dye-transfer assay. Based on these findings, TCDD has both tumor initiating and promoting potential in human breast epithelial cells in culture. Also, treatment of MCF10A cells with the carcinogen 7,12-dimethylbenz[a]anthracene plus TCDD resulted in malignant cell transformation as revealed by increased anchorage-independent growth of exposed cells. Additional studies may be necessary to assess the effects of TCDD on multi-stage carcinogenesis in vivo.

• Microbiology and Biotechnology Letters
• /
• v.34 no.4
• /
• pp.357-362
• /
• 2006

A growth inhibition assay using Chlorella sp. AG 10002 based on the OECD 201 standard test procedure was applied to the toxicity testing of endosulfan and its reported metabolites. Comparison of dry cell weight, optical density (OD) at 680 nm, and chlorophyll a concentration indicated that optical density at 680 nm of culture broth is convenient, rapid, and accurate method for cell growth. In this microalgae system, the $IC_{50}$ values of endosulfan, endosulfan sulfate, endosulfan lactone, and endosulfan ether were determined as 9.45, 18.8, 18.2 and 37.5 mg/L, respectively. In a while, endosulfan diol did not show a significant toxicity up to 50 mg/L. Since endosulfan is liable at acidic or alkaline conditions, treatment of endosulfan in pH 3, 4, and 11 for 3 days resulted in reduced toxicity, as expected. These results suggested that the microalgae system is useful to evaluate various toxic chemicals and provide a new notion for bioremediation of endosulfan in aqueous systems.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.13
• /
• pp.5325-5329
• /
• 2015

Farnesiferol C is a natural compound with various anti-cancer properties that belongs to the class of sesquiterpene coumarins. However, the low bioavailability of farnesiferol C limits its therapeutic potential. Here, we overcame this problem utilizing dendrosome nano-particles and evaluated the anti-cancer effect of dendrosomal farnesiferol C (DFC) on the AGS gastric cancer cell line. The 3-(4,5-dimethyl-thiazol-2yl)-2,5- diphenyl tetrazolium bromide (MTT) assay and reverse transcriptase-polymerase chain reaction (RT-PCR) were respectively used to detect the anti-proliferative properties of DFC and expression ratio of Bax/Bcl-2 as a hallmark of apoptosis. Compared to the void farnesiferol C (FC), our data showed that DFC significantly suppresses the proliferation of AGS cells in a time- and dose-dependent manner (P<0.01). Also, DFC meaningfully increased the expression ratio of Bax/Bcl-2 in AGS cells (P<0.01). The findings demonstrate that our nano-based formulation of farnesiferol C could be considered as a potential therapeutic agent in cancer targeting.

• Applied Microscopy
• /
• v.5 no.1
• /
• pp.31-43
• /
• 1975

These studies were carried out to detect the presence of infected virus- like particles and also were observed the ultrastructures of Aspergillus ochraseus isolated from kokja and Korean ginseng. The results of ultrastructures of Aspergillus ochraseus are summarized as follows: 1. In fungal cells, nuclei were enclosed by a irregular double membrane and nucleoli in the nucleus. 2. In cytoplasm, mitochondria, rough endoplasmic reticulum with ribosomes and glycogen were scattering distributed and many lomasomes also observed. 3. The osmiophilic bodies of fungal cells existed in the vesicles. 4. The cell walls were composed of a low electron dense materials. 5, Conidia cell walls were extremely thick and possessed the high electron density of outer coat. The virus-like particles were observed in the hyphae of Penicillium chrysogenum Q-176. These virus-like particles measured $350{\AA}$ in diameter. But strains of Aspergillus ochraseus, showing some vesicle particles were also observed about $800{\AA}$ in diameter in the central region of young fungal hyphae. Based on the results of these experiments, it can not be determined virus particles or not. The further studies to determination of virus particles will be proceeded by the chemical, physical and biological assay methods.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.10
• /
• pp.1613-1621
• /
• 2006

Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of ${\alpha}-\;and\;{\beta}$-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.

• Mycobiology
• /
• v.45 no.4
• /
• pp.385-391
• /
• 2017

The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.