• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.3701-3704
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• 2012

Background: Cervical cancer is one of the most common cancers in developing countries and the second most common type of cancer in women globally. Several recent studies suggested a co factor role for Chlamydia trachomatis in pathogenesis of cervical cancer. This study aimed to evaluate existence of C. trachomatis DNA in pathologic blocks of patients with cervical cancer. Materials and methods: Seventy-six formaldehyde fixed paraffin embedded tissue specimens from patients with histologically proven history of cervical cancer as well as 150 blocks from healthy peoples were included in the present study. Thin slices were prepared from selected blocks followed by deparaffinization and DNA extraction; the presence of C. trachomatis DNA was examined by Taq Man real-time PCR. Results: Our TaqMan real time PCR assay with cervical specimens of patients with cervical cancer showed that there was no C. trachomatis DNA. Also, we found three positive specimens among our control group. Conclusion: It seems that based on results obtained from the specimens examined in the present study, there is no association between the presence of C. trachomatis DNA in cervical specimens and cervical cancer.

• Proceedings of the Korean Society of Life Science Conference
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• 2006.09a
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• pp.66.1-66.1
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• 2006

• Journal of Veterinary Clinics
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• v.34 no.5
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• pp.388-391
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• 2017

A 8-month-old, spayed female, Domestic shorthair cat lived in a shelter was presented with pelvic limbs ataxia and dysuria. Serum biochemical profile abnormalities were hyperproteinemia and decreased albumin/globulin (A:G) ratio (0.70). Results of cerebrospinal fluid (CSF) analysis were mixed cells pleocytosis with predominance neutrophils and an increase in protein concentration. In addition, feline coronavirus was detected by realtime RT-PCR in CSF. Magnetic resonance imaging (MRI) findings revealed lesions of the lumbar spinal cord. Based on clinical signs, MR finding, CSF analysis and realtime RT-PCR result in CSF, this case was diagnosed as feline infectious peritonitis (FIP) associated meningomyelitis. Although prednisolone and mycophenolate mofetil were administrated, clinical signs were not resolved and progressed to tetraplegia and coma status. This case presentation describes that feline infectious peritonitis virus could affect the lumbar spinal cord only and cause meningomyelitis with pelvic limbs ataxia without other neurological signs.

• Biomedical Science Letters
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• v.15 no.4
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• pp.319-326
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• 2009

Extracellular ATP elicits diverse physiological effects by binding to the G-protein-coupled P2Y receptors on the plasma membrane. In addition to the short-term effects of extracellular nucleotides on cell functions, there is evidence that such purinergic signalling can have long-term effects on cell proliferation, differentiation and death. The 3T3-L1 cell line derived from mouse embryo is a well-established and commonly utilized in vitro model for adipocytes differentiation and function. However, the distributions and roles of P2Y subtypes are still unknown in the preadipocyte. In this study, we identified the distributions and roles of P2Y subtypes in preadipocyte using $Ca^{2+}$ imaging and realtime PCR. ATP increased the $[Ca^{2+}]_i$ in a concentration-dependent manner. ATP increased $Ca^{2+}$ in absence and/or presence of extracellular $Ca^{2+}$. Suramin, non-selective P2Y blocker, largely blocked the ATP-induced $Ca^{2+}$ response. U73122, a PLC inhibitor, completely inhibited $Ca^{2+}$ mobilization in 3T3-L1 cells. The mRNA expression by realtime PCR of P2Y subtypes was $P2Y_2:P2Y_5:P2Y_6=1.0:12.5:0.3$. In conclusion, we showed that $P2Y_5$ receptor is a dominant purinergic receptor in preadipocytes, and multiple P2Y receptors could involve in differentiation and migration via regulating of intracellular calcium concentration.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.569-573
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• 2010

A quantitative detection method for Salmonella in seafood was developed using a SYBR Green-based real-time PCR assay. The assay was developed using pure Salmonella DNA at different dilution levels [i.e., 1,000 to 2 genome equivalents (GE)]. The sensitivity of the real-time assay for Salmonella in seeded seafood samples was determined, and the minimum detection level was 20 CFU/g, whereas a detection level of 2 CFU/ml was obtained for pure culture in water with an efficiency of ${\geq}85%$. The real-time assay was evaluated in repeated experiments with seeded seafood samples and the regression coefficient ($R^2$) values were calculated. The performance of the real-time assay was further assessed with naturally contaminated seafood samples, where 4 out of 9 seafood samples tested positive for Salmonella and harbored cells <100 GE/g, which were not detected by direct plating on Salmonella Chromagar media. Thus, the method developed here will be useful for the rapid quantification of Salmonella in seafood, as the assay can be completed within 2-3 h. In addition, with the ability to detect a low number of Salmonella cells in seafood, this proposed method can be used to generate quantitative data on Salmonella in seafood, facilitating the implementation of control measures for Salmonella contamination in seafood at harvest and post-harvest levels.

• Journal of Mushroom
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• v.9 no.2
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• pp.53-58
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• 2011

In order to confirm the presence of putative glucoamylase gene in Tricholoma matsutake genome, the genomic DNA was prepared from T. matsutake NBRC30773 strain and was used as template to clone the glucoamylases gene (TmGlu1). We obtained the nucleotide sequence of TmGlu1 and its franking region. The coding region (from ATG to stop codon) is 2,186 bp. The locations of exons and introns were determined from the nucleotide sequences of 3'- and 5'-RACE PCR and RT-PCR products. On the other hand, to investigate the relationship between composition of medium and glucoamylase expression, we checked the expression level of glucoamylase gene by realtime reverse transcription PCR and measurement of glucoamylase enzyme activity. It was found that enzyme activity of glucoamylase was very low in different medium. Expression of glucoamylases gene appeared to not be affected by different carbon source.

• Journal of Ginseng Research
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• v.34 no.1
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• pp.47-50
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• 2010

The common DNA extraction methods are indispensable for genotyping by molecular marker analysis. However, genotyping a large number of plants is painstaking. A modified 'NaOH-Tris' method used in this study reduces the extraction time while keeping the cost low and avoiding the use of hazardous chemicals. The endpoint analysis by realtime PCR tends to be fast and effective for the development of SNP markers linked to the 'Chunpoong' cultivar of Panax ginseng. The 'Chunpoong' marker was developed by a major latex-like protein gene sequence. From our results, we suggest that this method is successful in distinguishing 'Chunpoong' from a large number of ginseng cultivars.

• Korean Journal of Pharmacognosy
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• v.45 no.2
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• pp.168-173
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• 2014

The aim of this study was to evaluate the anti-helicobacter activity of the ethanol extract of Sohamhyoongtang (Coptidis Rhizoma, Pinelliae Tuber and Trichosanthis Semen) and Coptidis Rhizoma total alkaloids, which is one of the components of Sohamhyoongtang. Crude ethanol extract of Sohamhyoongtang (ESHHT) and Coptidis Rhizoma total alkaloids (CRTA) were used for this experiment. Five different types of H. pylori (including H. pylori 26695) were used as test strain. To determine anti-helicobacter activity, minimum inhibitory concentration (MIC) was determined by agar dilution method. The effect of ESHHT and CRTA on the gene expression of H. pylori was investigated by quantitative realtime-PCR (qRT-PCR). MICs of ESHHT against five H. pylori strains were $250{\sim}500{\mu}g/ml$ and MICs of CRTA against five H. pylori strains were $50{\sim}200{\mu}g/ml$. Four representative virulence genes of H. pylori, cagA, ureA, ureB and ureI were tested as target genes for qRT-PCR. According to the qRT-PCR results, both ESHHT and CRTA markedly repressed the expression of cagA gene of H. pylori 26695 (6.91 and 20 folds respecively). These results showed that the ESHHT and CRTA demonstrated antihelicobacter properties, suggesting their potential use in gastritis or duodenal ulcer.

• Korean Journal of Food Science and Technology
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• v.53 no.1
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• pp.1-11
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• 2021

In this review, recent foodborne outbreaks caused by viruses in the Republic of Korea (2010-2019) were analyzed. The human norovirus was found to be the major foodborne virus causing an average of 94.9% of the viral outbreaks. Reverse-transcription polymerase chain reaction (PCR) with electrophoresis has been widely used to detect viruses, but several rapid detection methods, including real-time PCR, multiplex PCR, and quantum dot assay, have also been suggested. For norovirus inactivation studies, surrogates such as murine norovirus and feline calicivirus have been widely used to identify the reduction rate owing to the limitations in laboratory cultivation. Conversely, direct cell infection studies have been conducted for other foodborne viruses such as adenovirus, astrovirus, rotavirus, and hepatitis A or E virus. Moreover, virucidal mechanisms using various physical and chemical treatments have been revealed. These recent studies suggest that rapid in situ detection and effective control are valuable for ensuring food safety against viral infections.

• BMB Reports
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• v.41 no.9
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• pp.670-677
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• 2008

A cDNA microarray composed of 2,028 different ESTs from two shrimp species, Penaeus monodon and Masupenaeus japonicus, was employed to identify yellow head virus (YHV)-responsive genes in hemocytes of P. monodon. A total of 105 differentially expressed genes were identified and grouped into five different clusters according to their expression patterns. One of these clusters, which comprised five genes including cathepsin L-like cysteine peptidase, hypothetical proteins and unknown genes, was of particular interest because the transcripts increased rapidly ($\leq$ 0.25 hours) and reached high expression levels in response to YHV injection. Microarray data were validated by realtime RT-PCR analyses of selected differentially expressed transcripts. In addition, comparative analysis of the hemocyte transcription levels of three of these genes between surviving and non-surviving shrimp revealed significantly higher expression levels in surviving shrimp.