• 한국어류학회지
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• 제9권1호
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• pp.91-98
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• 1997

외래유전자, pFV4CAT이 이식된 transgenic 미꾸라지 계통 F1 및 F2를 대상으로 조직별 외래 유전자의 발현을 조사하였다. Transgenic F1에서 pFV4CAT의 mRNA 합성 여부를 조사하기 위해 정소(testis), 간(liver), 근육(muscle), 비장(spleen) 및 심장(heart) 조직을 RT-PCR로 분석한 결과, 조직별 mRNA 존재 여부는 F1 계통간 큰 차이를 나타내었으며, 다른 조직들에 비해 간(liver)과 비장(spleen)에서 보다 빈번히 발현하는 경향을 나타내었다. 외래 유전자에 의해 합성된 CAT 단백질을 ELISA로 정량화한 결과, 조직별 및 transgenic 계통별 다양한 차이가 있었으며, 다른 조직에 비해 근육과 심장에서 가장 높은 수준으로 발현하는 것으로 나타나 F1 한 계통의 근육에서, 대조군 수치의 최고 68배에 해당하는 CAT 발현이 관찰되였다. 반면 정소에서 가장 낮은 외래 유전자의 발현이 모든 transgenic line에서 관찰되었다.

• 한국임상수의학회지
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• 제33권3호
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• pp.145-150
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• 2016

We determined the prevalence of feline hemotropic mycoplasma species including 'Candidatus Mycoplasma haemominutum', Mycoplasma haemofelis, and 'Candidatus Mycoplasma turicensis' in naturally infected feral cats in Jeonju, Korea. Forty six feral cats were evaluated by PCR assay targeting the 16S rRNA gene sequence. Nine cats (19.6%) were positive for 'Candidatus Mycoplasma haemominutum', 2 cats (4.3%) were positive for 'Mycoplasm a haemofelis', and 1 cat (2.2%) was infected with both 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis. 'Candidatus Mycoplasma turicensis' was undetected. Partial 16S rRNA gene sequences of Mycoplasma haemofelis were closely (> 96%) related to those from other countries. The amplification of hemoplasma DNA in these samples confirmed the presence of 'Candidatus M. haemominutum' and M. haemofelis in Korea.

• Journal of Plant Biotechnology
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• 제36권2호
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• pp.149-156
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• 2009

Application of exogenous polyamines (PAs) reduced the paraquat (PQ)-induced cotyledon injuries in radish seedling plants with 1 mM spermidine (Spd) being the most effective protectant. PQ injury symptoms in the cotyledons, e.g., large accumulation of $H_2O_2$, and losses of fresh weight, chlorophyll, and proteins, were significantly alleviated. Likewise, analysis of $H_2O_2$-scavenging enzymes such as catalase (CAT) and guaiacol peroxidase (GPX) showed that pretreatment with Spd among PAs remarkably increased total CAT activity and strongly retarded PQ-induced rapid decline in total GPX activity. In a native gel assay, one CAT isozyme (CAT1) and two GPX isozymes (GPX1 and a newly synthesized GPX isozyme) proved to be more responsible for PQ tolerance, as manifested by the strong increases in their activities by Spd pretreatment. Based on these results, we can suggest that PAs (especially 1 mM Spd) may function as antioxidant protectors by invoking CAT and GPX enzymes which control the endogenous $H_2O_2$ level in radish cotyledons exposed to PQ.

• Journal of Veterinary Science
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• 제22권6호
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• pp.67.1-67.7
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• 2021

Background: Chlamydophila felis, formerly known as Chlamydia psittaci var. felis, is frequently associated with ocular, respiratory, and occasionally reproduction tract infections. Even though the infection is sometimes asymptomatic, it potentially results in a latent immunosuppressive infection. Objective: This study aimed to identify occurrences of feline chlamydophilosis, rarely reported in cats in Indonesia. Methods: The observation was conducted in three cats with clinical signs of Cp. felis infection, particularly relapsing conjunctivitis. The cats' histories were recorded based on owners' information. Conjunctival swabs were sampled for cytology examination and molecular assay detection. A phylogenetic tree was generated using MEGA-X software to reveal group clustering. A post-mortem examination was performed on the cat that died during an examination. Results: Cp. felis was detected in both cytological examination and polymerase chain reaction assay. The phylogenetic tree demonstrated that the Cp. felis isolated in this study clustered with several other isolates from the other countries. Cp. felis can be isolated from cats with different clinical manifestations and levels of severity. The chronic fatal infection demonstrated interstitial broncho-pneumonia under histopathological examination. Conclusions: Molecular assay of Cp. felis is always recommended to obtain a definitive diagnosis of feline chlamydophilosis since the disease can have various clinical manifestations. Even though it may be subclinical and is often not fatal, an infected cat may be a carrier that could spread the pathogen in the surrounding environment. Serious disease management is suggested to avoid high costs associated with regularly relapsing disease.

• 한국환경보건학회지
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• 제34권5호
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• pp.369-373
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• 2008

The indoor environment is an important source of exposure to various aeroallergens and pathogenic microorganism. It has been shown that exposure to aeroallergens enhances the risk of indoor inhabitants developing asthma. Since the skin prick test, a typical clinical method for identification of subjects positive to allergens, can rarely cause fatal or non-fatal reactions in susceptible persons, an in vitro assay such as ELISA using serum has been considered for testing positivity against various allergens. We evaluated the validity of a serum ELISA kit for screening positive subjects to major aeroallergens including Dermatophagoides farinae, Dermatophagoides pteronyssinus, cockroach, Alternaria, Cladosporium, Aspergillus, Penicillium, dog hair, cat fur, mugwort, and ragweed. The ELISA results were compared with the skin prick test results, and sensitivity, specificity, and overall accuracy were calculated to each allergen. Higher sensitivities were obtained from D. farinae, (77.8%) and D. pteronyssinus (69.2%), but sensitivities to Aspergillus, Penicillium, dog hair, cat fur, and ragweed were very low down to 0%. Specificity ranged from 88.7% (cat fur) to 100% (mugwort and ragweed). Overall the accuracy of the serum ELISA kit was relatively high, in that the lowest was 85.1% for cat fur and the highest was 98.6% for Alternaria, Cladosporium, and ragweed. Considering specificity and overall accuracy for the serum ELISA kit, it may be considered reliable. However, when the kit is used for screening purpose, positivity to aeroallergens should be carefully determined since sensitivity for the kit is low.

• Archives of Pharmacal Research
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• 제27권9호
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• pp.906-911
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• 2004

In order to evaluate estrogenic compounds in natural products, an in vitro detection system was established. For this system, the human breast cancer cell line MCF7 was stably trans-fected using an estrogen responsive chloramphenicol acetyltransferase (CAT) reporter plas-mid yielding MCF7/pDsCAT-ERE119-Ad2MLP cells. To test the estrogenic responsiveness of this in vitro assay system, MCF7/pDsCAT-ERE119-Ad2MLP cells were treated with various concentrations of 17f3-estradiol. Treatments of 10$^{-8}$ to 10$^{-12}$ M 17$\beta$-estradiol revealed significant concentration dependent estrogenic activities compared with ethanol. We used in vitro assay system to detect estrogenic effects in Puerariae radix and Ginseng radix Rubra extracts. Treat-ment of 500 and 50 $\mu\textrm{g}$/ml of Puerariae radix extracts increased the transcriptional activity approximately 4- and 1.5-fold, respectively, compared with the ethanol treatment. Treatment of 500, 50, and 5 $\mu\textrm{g}$/ml of Ginseng radix Rubra extracts increased the transcriptional activity approximately 3.2-,2.7, and 1.4-fold, respectively, compared with the ethanol treatment. These observations suggest that Puerariae radix and Ginseng radix Rubra extracts have effective estrogenic actions and that they could be developed as estrogenic supplements.

• 혜화의학회지
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• 제23권2호
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• pp.5-13
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• 2015

Objectives : The anti-inflammatory and antioxidant effects of water extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) were investigated. The effects of SSJ and GSJ were compared. Methods : We performed cell viability assay in HaCaT cells and RAW 264.7 cells using 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide(MTT) assay. We measured chemokines(regulated on activation normal T-cell expression and secreted ; RANTES/CCL5, interferon-inducible protein; IP-10/CXCL10, macrophage-derived chemokine; MDC/CCL22) in HaCat cells, also we measured cytokines (tumor necrosis factor-${\alpha}$; TNF-${\alpha}$, interleukin-6; IL-6) and nitric oxide(NO) production in RAW 264.7 cells using enzyme-linked immunosorbent assay(ELISA) and NO assay. Western blot assay was used to evaluate the expression for inducible nitric oxide synthase(iNOS) in RAW 264.7 cells. Results : SSJ and GSJ did not affect the cell viability at the concentrations treated ($0-800{\mu}g/ml$). As a result of SSJ and GSJ treatment in HaCat cells stimulated by TNF-${\alpha}$(10 ng/ml) and interferon(IFN)-${\gamma}$(10 ng/ml), the production of RANTES and IP-10 was inhibited significantly. However there was no significant difference in the secretion of MDC. And in RAW 264. 7 cells stimulated by lipopolysaccharide(LPS, $1{\mu}g/ml$), SSJ and GSJ treatment significantly inhibited the secretion of TNF-${\alpha}$ and IL-6 and the production of NO. The expression of iNOS was also decresed by SSJ and GSJ treatment in RAW 264. 7 cells. Compared with SSJ, GSJ was superior to SSJ in inhibition of RANTES, IP-10, TNF-${\alpha}$, IL-6 and NO production at the concentration of $200{\mu}g/ml$. Conclusion : Both SSJ and GSJ have anti-inflammatory and antioxidant effects. And GSJ has better effects than SSJ.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.895-901
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• 2013

Resistance to induction of apoptosis is a major obstacle for bladder cancer treatment. Bcl-2 is thought to be involved in anti-apoptotic signaling. In this study, we investigated the effect of Bcl-2 overexpression on apoptotic resistance and intracellular reactive oxygen species (ROS) generation in bladder cancer cells. A stable Bcl-2 overexpression cell line, BIU87-Bcl-2, was constructed from human bladder cancer cell line BIU87 by transfecting recombinant Bcl-2 [pcDNA3.1(+)-Bcl-2]. The sensitivity of transfected cells to adriamycin (ADR) was assessed by MTT assay. Apoptosis was examined by flow cytometry and acridine orange fluorescence staining. Intracellular ROS was determined using flow cytometry, and the activities of superoxide dismutase (SOD) and catalase (CAT) were also investigated by the xanthinoxidase and visible radiation methods using SOD and CAT detection kits. The susceptibility of BIU87-Bcl-2 cells to ADR treatment was significantly decreased as compared with control BIU87 cells. Enhanced expression of Bcl-2 inhibited intracellular ROS generation following ADR treatment. Moreover, the suppression of SOD and CAT activity induced by ADR treatment was blocked in the BIU87-Bcl-2 case but not in their parental cells. The overexpression of Bcl-2 renders human bladder cancer cells resistant to ADR-induced apoptosis and ROS might act as an important secondary messenger in this process.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.55-70
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• 2005

Objective: Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammnatory in the traditional Korean medicine. UD has been reported as a good enhancer for bone healing. Methods : In this experiment, we investigate the Inhibitory effects of UD on bone resorption using the bone cells culture. Different concentrations of crude extract of UD were added to mouse bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric MIT assay. It was demonstrated that UD has potential effects on bone cells culture without any cytotoxicity. The most effective concentration of UD on bone cells were $100\;{\mu}g/ml$. Cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. Results : When mouse long bone cells including osteoclasts and osteoblast were treated with the PI3-Kinase inhibitor, wortmannin (WT), WT prevented the osteoclast-mediated intracellular processing of Cat K. Similarly, treatment of osteoclasts-containing long bone cells with UD extracts prevented the intracellular maturation of Cat K, suggesting that UD may disrupt the intracellular trafficking of pro Cat K. This is similar to that of WT. Since secreted proenzymes have the potential to reenter the cell via mannose-6-phosphate (M6P) receptor, to prevent this possibility, we tested WT and UD in the absence or presence of M6P. Inhibition of Cat K processing by WT or UD was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of WT and UD. Conclusion : UD dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing.

• 한국안광학회지
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• 제20권2호
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• pp.237-246
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• 2015

목적: 본 연구는 안구에 존재하는 항산화효소인 superoxide dismutase(SOD)와 catalase(CAT)가 UV-B에 반복적으로 노출되었을 때 유발되는 구조변성 및 활성저하의 상관관계를 밝히며, 자외선 차단 RGP렌즈의 사용으로 효소의 변성과 활성저하가 효율적으로 차단되는가를 알아보기 위하여 수행되었다. 방법: SOD와 CAT의 표준품으로 각각의 효소용액을 제조하고 하루 30분, 1시간 및 2시간씩 312 nm의 UV-B에 1, 2, 3, 4 및 5일 동안 반복적으로 노출시켰다. 이 때 UV-B에 직접 노출시킨 항산화효소의 구조 및 활성변화를 자외선 차단기능이 있는 RGP렌즈로 UV-B를 차단시킨 경우와 비교하였다. UV-B 반복노출에 따른 SOD와 CAT의 구조변성은 전기영동분석으로 확인하였으며, 이들 효소활성은 분석키트를 이용하여 비색분석법으로 측정하였다. 결과: UV-B에 반복노출된 SOD는 일일 30분 조사조건으로 반복노출되었을 때에도 전기영동분석에서 효소다중화(polymerization)가 관찰되었으나 활성의 변화는 10% 이내로 나타났다. 반면 UV-B에 반복노출된 CAT은 전기영동 시 효소밴드크기나 진하기가 감소하여 구조변성이 나타났음을 확인할 수 있었으며, 반복노출시간이 긴 경우 CAT은 전기영동분석에서 효소밴드를 보임에도 불구하고 그 활성은 완전히 소실되었다. 또한 UV-B 조사로 인한 CAT의 변성은 63.7%의 UV-B 차단기능을 가진 RGP렌즈의 사용으로 어느 정도 억제되었으나 완전히 억제되는 것은 아니었다. 결론: 이상의 결과로 UV-B 반복노출에 따른 항산화효소의 구조변성은 그 종류에 따라 효소활성의 감소정도와 반드시 일치하는 것은 아님을 알 수 있었다. 따라서 자외선으로 인한 항산화효소의 손상을 막기 위하여서는 콘택트렌즈를 착용한 상태에서 자외선 노출시간을 최소화하거나, FDA Class I 차단제에 해당하는 UV 차단율을 가지는 콘택트렌즈를 착용 또는 이에 상응하는 UV차단율을 가지는 선글라스를 덧착용할 것을 권장한다.