• Parasites, Hosts and Diseases
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• 제56권1호
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• pp.87-91
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• 2018

Morphological and molecular characteristics of spirometrid tapeworms, Spirometra decipiens, were studied, which were recovered from a heavily infected stray cat road-killed in Eumseong-gun, Chungcheongbuk-do (Province), the Republic of Korea (=Korea). A total of 134 scolices and many broken immature and mature proglottids of Spirometra tapeworms were collected from the small intestine of the cat. Morphological observations were based on 116 specimens. The scolex was 22.8-32.6 mm (27.4 mm in average) in length and small spoon-shape with 2 distinct bothria. The uterus was coiled 3-4 times, the end of the uterus was ball-shaped, and the vaginal aperture shaped as a crescent moon was closer to the cirrus aperture than to the uterine aperture. PCR amplification and direct sequencing of the cox1 target fragment (377 bp in length and corresponding to positions 769-1,146 bp of the cox1 gene) were performed using total genomic DNA extracted from 134 specimens. The cox1 sequences (377 bp) of the specimens showed 99.0% similarity to the reference sequence of S. decipiens and 89.3% similarity to the reference sequence of S. erinaceieuropaei. In the present study, we report a stray cat heavily infected with S. decipiens identified by mitochondrial cox1 sequence analysis and morphological examinations of the adult worms.

• Asian-Australasian Journal of Animal Sciences
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• 제23권7호
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• pp.855-862
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• 2010

Among the known interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful pathways. The Mx protein has direct antiviral activity and inhibits a wide range of viruses by blocking an early stage of the viral replication cycle. Cloning, characterization, and expression of Mx in vivo and in vitro have been conducted. The chicken Mx gene spans 21 kb and is made up of 14 exons and 13 introns, of which the promoter region was analyzed. The real-time PCR results showed that Mx expression was increased in chicken embryo fibroblasts (CEF) after 12- and 24-h induction with polyI: C. Induction of Mx expression by poly I: C in vivo revealed tissue-specific patterns among the chicken tissues tested. A trace expression of Mx was detected in healthy chicken liver tissues from adult chickens without inducement; the expression levels in the liver, heart, and gizzard were higher than in the muscle and kidney. This is the first report to demonstrate the expression of a glutathione-S-transferase-tagged-Mx fusion protein of 75 KDa, as well as the biological activity tested by SDS-PAGE and western blotting.

• 한국잠사곤충학회지
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• 제40권1호
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• pp.52-59
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• 1998

For the construction of plasmid and bmNPV sarrying the FRT recognition site for the FLP recombinases, we synthesized the wild type FRT dligonucleotides. The target FRT sequences consist of three 13bp repeated DNA sequences; two repeats in a direct orientation and one inverted relative to the other two. In addition, there is an 8bp spacer region between the repeats which determune the orientation of the FRT recombination site. In order to place the FRT site both in target BmNPV genome and the transfer vector, we constructed a plasmid, FRT site both in the target BmNPv genome and the transfer vector, we constructed a plasmid, pFRT$\beta$-gal, carrying the FRT sites within the cloning sites of pSV vector and a recombinant BmNPV, vFRTPH, carrying the FRT sites at a downstream of polyhedrin promotor, respectively. In order to test the functionality of the FLP/FRT site-specific recombination system, vFRTPH, pFRT$\beta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$\beta$2-gal. From construction analysis of the vFRT$\beta$2-gal with PCR technique it was concluded that the entire pFRT$\beta$-gal plasmid with $\beta$-galactosidase gene and origines of replication flanked by two functional hybrid FRT sequences. The efficiency of recombination was 8.7%, which was higher than that(2.2%) of recombination between a conventional transfer vector and the wild type BmNPV.

• Clinical and Experimental Pediatrics
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• 제46권5호
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• pp.505-509
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• 2003

저자들은 빈번한 구토와 성장 장애, 발달 지연 등을 주소로 내원한 1년 7개월된 남아에서 HPRT에 대한 생화학적 효소 분석을 통해 진단하고, HPRT 유전자에 대한 분자유전학적 분석을 시행하여 병인이 되는 돌연변이가 확인된 Lesch-Nyhan 증후군 1례를 경험하였기에 이를 문헌고찰과 함께 보고하는 바이다.

• 대한한방내과학회지
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• 제28권4호
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• pp.681-693
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• 2007

Background and Objective : Atractylodes japonica (AJ) is a commonly-used herbal medicine in Asian countries such as Korea, China and Japan. The present study was designated to evaluate the direct effects of AJ on helper T cell activities and on Th1/Th2 lineage development in vitro. Materials and Methods : Spleen cells from 8-week BALB/c mice were cultured in CR extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and analyzed for mRNA expression levels of INF-$\gamma$, IL-4, T-bet and GATA-3 by RT-PCR and secretion cytokines levels of INF-$\gamma$, IL-2, IL-4, IL-5 and IL-10 by ELISA. Results : The results demonstrated that AJ had no mitogenic effects on unstimulated CD4+ T cells, but augmented CD4+T-cell proliferation upon activation with anti-CD3/anti-CD28 antibodies in a dose-dependent manner. AJ treatment significantly increased CD4+ T cell population and IFN-$\gamma$ expression was significantly enhanced, while IL-4 expression significantly decreased. In addition, in vitro Th1/Th2 polarization experiments revealed that AJ enhanced IFN-$\gamma$ secretion in Th1 cells, but reduced the IL-4 in Th2 cells in dose-dependent manner. Conclusion : These results suggest that AJ treatment could be a desirable alternative therapy for the prevention or correction of Th2 dominant pathological disorders, such as allergy and asthma.

• 동의생리병리학회지
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• 제16권4호
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• pp.693-700
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• 2002

SCRT (Sochungyong-tang) has been used for immune disease in human. The purpose of this study was effect of Helper T cell, major regulator of immune system. Spleen cell from 8 week BALB/c mice were cultured in SCRT containing medium without activation for 48 h. The MTS assay and flow cytometry revealed that SCRT treated Iympocyte were non-effect in percentage of CD4+ T cell. Subsequently CD4+ T cell were isolated and cultured in SCRT containing medium. SCRT were non-effective on CD4+ T cell without any involvement of APC. In order to evaluate the direct effect of SCRT on Helper T cell, CD4+ T cell isolated after 48 h of culture in SCRT containing medium and activated with and without anti-CD3/anti-CD28 activation for 48 h. A lower level of CD69 was observed in SCRT treated cells in flow cytometry analysis. Subsequently Using RT-PCR analysis the expression of mRNA for IL-2, INF-γ are upregulated and, IL-4 is downregulated in CD4 T cell. The result suggests that SCRT makes Th1 significantly increased and Th2 relatively inhibited. The results suggest that SCRT potentiate Th1 cell and decrease Th2 development at the same time, which is believed to be bemeficial for IgE-mediated responses.

• 동의생리병리학회지
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• 제27권6호
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• pp.745-751
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• 2013

Safflower (Carthamus tinctorius L.) seeds have been used in Korea and China for promoting bone formation and protection. This study was designed to examine the Nrf-2 mediated anti-oxidative effects of Korean and Chinese safflower seeds. Water and ethanol extracts of safflower seeds were treated to RAW 264.7 cells. Nrf-2 transcriptional activity was measured by reporter gene assay and western blot analysis. Semi-quantitive RT-PCR analysis was adopted to measure Nrf-2 dependent gene expressions. Water extracts of safflower seeds have strongly induced the activation of Nrf-2 transcription than ethanol extracts. Especially, water extracts of Korean safflower seeds has more strongly increased the expression of nuclear Nrf-2. Water extracts of Korea and China safflower seeds have also increased the expression of Nrf-2-dependent genes such as GCLC, NQO-1 and HO-1 in RAW 264.7 cells. However, all kinds of safflower seeds extracts did not increase intracellular ROS production. These results demonstrate that the antioxidant effects of safflower seeds are not related with ROS production, rather it is mediated by the direct activation of Nrf-2.

• 한국발생생물학회지:발생과생식
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• 제14권1호
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• pp.35-41
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• 2010

Mammalian reproduction is regulated by a feedback circuit of the key reproductive hormones such as GnRH, gonadotropin and sex steroids on the hypothalamic-pituitary-gonadal axis. In particular, the onset of female puberty is triggered by gain of a pulsatile pattern and increment of GnRH secretion from hypothalamus. Previous studies including our own clearly demonstrated that genistein (GS), a phytoestrogenic isoflavone, altered the timing of puberty onset in female rats. However, the brain-specific actions of GS in female rats has not been explored yet. The present study was performed to examine the changes in the activities of GnRH neurons and their neural circuits by GS in female rats. Concerning the drug delivery route, intracerebroventricular (ICV) injection technique was employed to eliminate the unwanted actions on the extrabrain tissues which can be occurred if the testing drug is systemically administered. Adult female rats (PND 100, 210-230 g BW) were anaesthetized, treated with single dose of GS ($3.4{\mu}g$/animal), and sacrificed at 3 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly raised the transcriptional activities of enhanced at puberty1 (EAP-1, p<0.05), glutamic acid decarboxylase (GAD67, p<0.01) which are known to modulate GnRH secretion in the hypothalamus. However, GS infusion could not change the mRNA level of nitric oxide synthase 2 (NOS-2). GS administration significantly increased the mRNA levels of KiSS-1 (p<0.001), GPR54 (p<0.001), and GnRH (p<0.01) in the hypothalami, but decreased the mRNA levels of LH-$\beta$ (p<0.01) and FSH-$\beta$ (p<0.05) in the pituitaries. Taken together, the present study indicated that the acute exposure to GS could directly activate the hypothalamic GnRH modulating system, suggesting the GS's disrupting effects such as the early onset of puberty in immature female rats might be derived from premature activation of key reproduction related genes in hypothalamus-pituitary neuroendocrine circuit.

• Environmental Engineering Research
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• 제14권1호
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• pp.19-25
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• 2009

In this study, total bacteria, enteric members of the $\gamma$-proteobacteria, and microbial communities in seawater were analyzed as indirect indicators for quantifying biofouling. Biomass in seawater can significantly affect feed water pretreatment and membrane biofouling of reverse osmosis desalination processes. The purpose of this paper is to investigate microbiological quantity and quality of seawater at the potential intake of a desalination plant. For this analysis, the total direct cell count (TDC) using 4'-6-diamidino-2-phenylindole (DAPI)-staining and DNA-based real-time PCR were used to quantify the total bacteria and relative content of enteric members of $\gamma$-proteobacteria in seawater, respectively. In addition, microbial communities were examined using 16S rRNA gene cloning and bacterial isolation to identify the most abundant bacteria for a further biofouling study. The experimental results of this study identified about $10^6$ cells/mL of (total) bacteria, $10^5$ 16S rRNA gene copies/mL of enteric $\gamma$-proteobacteria, and the presence of more than 20 groups of bacteria.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.98-98
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• 2003

Direct gene transfer to mammalian tissues has significant potential for gene therapy and transgenesis. Liposome-mediated in vivo transfection has begun to gain attention as an alternative to viral vectors, and may also be a good mode of transfection in gene transfer. Interestingly, polymerized cationic liposomes are reported to be very stable in the bloods and efficient for in vivo gene transfer. To examine a possible gene delivery in vivo, we investigated the efficacy and safety of the liposome-mediated gene transfer using vein injection in chick or mouse as model animals. The number of injected pGFP-LacZ using either a commercial or home-made liposomes was 8 and 19 at 16 and 7 day of hatch, respectively. One of injected chick of each experiments was analyzed and the rest is being bred. In mouse, 4/22 showed expression of pGFP-LacZ but 8/22 showed no expression and the remaining animals are also being bred. After injection of liposome/pGFP-LacZ complex into wing vein of 7 or 16 day-old chick, pGFP-LacZ was detected in various tissues isolated from not only young chick but also old chick were turned out to possess. exogenous DNA. Transcripts and proteins of the transgene were also detected by RT-PCR or histochemical analysis, respectively. These results suggest that injected DNA were inserted to genome and produced mRNA and proteins in various tissues and may give an important tools for effective gene delivery in gene therapy or transgenesis.