• BMB Reports
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• v.36 no.2
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• pp.173-178
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• 2003

Malignant melanoma is one of the most rapidly increasing cancer types, and patients with metastatic disease have a very poor prognosis. Detection of metastatic melanoma cells in circulation may aid the clinician in assessing tumor progression, metastatic potential, and response to therapy. Tyrosinase is a key enzyme in melanine biosynthesis. The gene is actively expressed in melanocytes and melanoma cells. Melan A is a differentiation antigen that is expressed in melanocytes. The presence of these molecules in blood is considered a marker for circulating melanoma cells. In this study, we analyzed the usefulness of this marker combination I evaluating the response to therapy in the blood of 30 patients with malignant melanoma. Circulating cells were detected by a reverse-transcriptase-polymerase-chain reaction. The tyrosinase expression was observed in 9 (30%) patients and Melan A in 19 (63.3%) patients before therapy. Following treatment, the tyrosinase mRNA was detected in only one patient, while Melan A transcripts were still present in 14 patients. We suggest that this molecular assay can identify circulating melanoma cells that express melanoma-associated antigens and may provide an early indication of therapy effectiveness.

• Korean Journal of Environmental Biology
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• v.31 no.3
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• pp.189-191
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• 2013

Mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) are widespread contaminants of food and feedstuffs. It is very likely, that humans and animals are always exposed to mixtures of mycotoxins rather than to individual compounds. Therefore, risk assessments should consider mixture toxicity data. In the present study the combination of AFB1, OTA and ZEA was tested for genotoxicity in rat bone marrow and blood leukocytes after 15, 30 and 60 days treatment. The level of DNA damage was determined by the comet assay. The tail intensity and Olive tail moment in leukocytes and bone marrow cells were significantly higher than in controls. At the same time, the level of DNA damage in bone marrow cells was higher than in leukocytes. The data suggests that prolonged exposure to mycotoxins combination through food consumption can induce DNA damage contributing to the harmful effects in vivo.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.794-797
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• 1999

Lactobacillus acidophilus is known to have an inhibitory activity on growth of Helicobacter pylori and this activity has been attributed to lactic acid and antibacterial agents produced by Lactobacillus. Since every Lactobacilli produces lactic acid, an another factor must exist for L. acidophilus to inhibit H. pylori growth. In this work, the inhibitory activity of L. acidophilus on H. pylori adherence was studied. An immunoabsorbent assay using TLC plate was developed and used for screening the inhibitory activity of various Lactobacilli on H. pylori adherence. Glycolipid, the attachment site for H. pylori, was isolated from blood type O red blood cells and spotted on a TLC plate. The H. pylori adherence increased linearly with increasing amounts of glycolipid spotted on the TLC plate. Various L. acidophilus strains, but not L. casei, appeared to inhibit H. pylori adherence to glycolipid, and the adherence decreased linearly as the concentration of the Lactobacillus increased. The results show that the inhibitory activity of L. acidophilus on H. pylori adherence is an another factor for L. acidophilus to inhibit H. pylori growth.

• The Korean Journal of Pesticide Science
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• v.8 no.2
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• pp.95-102
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• 2004

The present study was conducted to test and evaluate estrogenic effect of 17 pesticides including benomy1 and carbaryl, being suspected as endocrine disrupting chemicals. For estrogenic effect examination, luciferase assay were achieved with human ovarian cancer cell, BG1Luc4E2. Estrogenic effects of cypermethrin, dicofol, endosulfan, esfenvalerate, and fenvalerate were observed at the concentration of $10^{-5}$ M by estrogen receptor binding assay. Relative luciferase potency and relative luciferase effects compared with $10^{-10}$ M 17 $\beta$-estradiol were $10^{-5}$, 56% for dicofol, and $10^{-5}$, 72% for endosulfan, respectively. Estimated maximum daily intake for pesticides was calculated from maximum residue limit of agricultural commodity and food consumption was 1.2298 mg/person. Theoretically calculated blood estrogen level from dietary intake for pesticides based on MRL in Korea, 3.075 ng/L was equivalent to 15% of estrogen concentration in normal blood, but practical monitoring data, 0.01938 ng/L was equal to 0.09693% of estrogen concentration in normal blood.

• Nutrition Research and Practice
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• v.8 no.4
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• pp.368-376
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• 2014

BACKGROUND/OBJECTIVES: The objectives of this study were to investigate the effects of lycopene on the migration, adhesion, tube formation capacity, and p38 mitogen-activated protein kinase (p38 MAPK) activity of endothelial progenitor cells (EPCs) cultivated with high glucose (HG) and as well as explore the mechanism behind the protective effects of lycopene on peripheral blood EPCs. MATERIALS/METHODS: Mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation. EPCs were identified after induction of cellular differentiation. Third generation EPCs were incubated with HG (33 mmol/L) or 10, 30, and $50{\mu}g/mL$ of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation and apoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performed by replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity was assayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylated and non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and $50{\mu}g/mL$ of lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantly lower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirm the causal relationship between MAPK inhibition and the protective effects of lycopene against HG-induced cellular injury, we treated cells with SB203580, a phosphorylation inhibitor. The inhibitor significantly inhibited HG-induced EPC injury. CONCLUSIONS: Lycopene promotes proliferation, migration, adhesion, and in vitro vasculogenesis capacity as well as reduces apoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-induced EPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-induced EPC injury by inhibiting p38 MAPKs.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.559-564
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• 1998

The polymerase chain reaction(PCR) assay was used for rapid diagnosis from blood and organ samples experimentally infected with Listeria monocytogenes. This method used a pair of primers based on a unique region in the 16S rRNA sequence of L monocytogenes. Procedure A was based on dilution of the blood sample followed by lysis of bacterial cell and direct analysis of the lysate with PCR. In artificially infected blood samples with L monocytogenes, it was possible to detect fewer than 40 cells per ml of blood. However, L monocytogenes was detected low rates on infected organs by the direct PCR. In procedure B, enrichment cultivation was used to increase numbers of bacteria before lysis and PCR. L monocytogenes was detected from 23 samples of 24 liver and spleen, respectively, and 18 samples of 24 blood were found to be positive by PCR on a subset of 72 organ samples, whereas L monocytogenes were detected on 63 organ samples in classical culture technique. It was required to analyze including enrichment steps were 6h and 18h on the procedure A and B, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.6
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• pp.671-677
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• 1996

Erythrocytes from three different animal species were used to determine mannose-sensitive hemagglutination (MSHA) and mannose-resistant hemagglutination (MRHA) of 755 isolates obtained from rectal swabs of healthy pig. In addition, colony hybridization using digoxigenin-dUTP labeled polynucleotide probes was performed for the detection of heat-stable and heat-labile enterotoxin genes carried by MRHA positive isolates. Of 755 strains, 9, 4 and 28 strains gave a positive MRHA with bovine, equine and pig erythrocytes, respectively. Of these isolates, 28 (3.7%) were characterized for positive MRHA by at least one blood. Seven isolates gave a positive MRHA with two kinds of blood. Three gave a positive MRHA with three kinds of blood. Twenty-eight strains, while positive in MRHA, yielded negative signals in the colony hybridization assay for the detection of heat-stable (STaI and STaII) and heat-labile (LT) enterotoxin genes in E. coli.

• Journal of Veterinary Clinics
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• v.14 no.2
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• pp.230-234
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• 1997

In vitro effect of cortisol on the proliferation of canine peripheral blood mononuclear cells (MNC) was examined. The MNC was isolated from peripheral blood by a gradient centrifugation with Picoll-Hypaque. The cell proliferation assayed using a noneradioactive 5-Bromo-2'-deoxy-uridine (BrdU) kit. The MNC proliferated well in response to either phrtobeRagg$]$utinin-p (PHA-P) or culture supernatant from MNC stimulated with PHA-p. However, these proliferative responses of MNC were not affected by addition of coitisol of 1 to 1,OOfl ng/ml. The addition of cortisol in MNC culture with either PHA-P or corture supernatBnt from MNC stimulated with PHA-P far 4 days wag not also influenced on the viabilities of cultured MNC. In conclusions it was able to assay the cell proliferation with BrdU instead of radioactive isotope e.g. tritiated thymidine (3H-TdR). These results suggested that cortisol does not at least influence on MNC proliferation in vitro.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.1-7
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• 1998

The present study was performed to evaluate circadian rhythm of blood nortisol concentration and change of blood cortisol concentration by various stress type in domestic cats. The ELISA kit for cortisol measurement was evaluated on the effective assay range, day to day variation, within-run variation and accuracy. The results were 1-1,000 ng/ ml, 0.7-5.9%, 0.9-4.5% and 98%, respectively. The circadian rhythm of serum cortisol concentration is presented in cats. Change of serum cortisol concentration was also examined with several stressors, stimulation with hamster as a prey, beep electronic sounds, vehicle transportation and unfamiliar environment. Unfamiliar environment showed most remarkable change in serum cortisol level.

• Safety and Health at Work
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• v.7 no.2
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• pp.156-160
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• 2016

Background: Workers in pesticide manufacturing industries are constantly exposed to pesticides. Genetic biomonitoring provides an early identification of potential cancer and genetic diseases in exposed populations. The objectives of this biomonitoring study were to assess DNA damage through comet assay in blood samples collected from industry workers and compare these results with those of classical analytical techniques used for complete blood count analysis. Methods: Samples from controls (n = 20) and exposed workers (n = 38) from an industrial area in Multan, Pakistan, were subjected to various tests. Malathion residues in blood samples were measured by gas chromatography. Results: The exposed workers who were employed in the pesticide manufacturing industry for a longer period (i.e., 13-25 years) had significantly higher DNA tail length ($7.04{\mu}m$) than the controls ($0.94{\mu}m$). Workers in the exposed group also had higher white blood cell and red blood cell counts, and lower levels of mean corpuscular hemoglobin (MCH), MCH concentration, and mean corpuscular volume in comparison with normal levels for these parameters. Malathion was not detected in the control group. However, in the exposed group, 72% of whole blood samples had malathion with a mean value of 0.14 mg/L (range 0.01-0.31 mg/L). Conclusion: We found a strong correlation ($R^2=0.91$) between DNA damage in terms of tail length and malathion concentration in blood. Intensive efforts and trainings are thus required to build awareness about safety practices and to change industrial workers' attitude to prevent harmful environmental and anthropogenic effects.