• Title/Summary/Keyword: IS-PCR

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Expression of Nesfatin-1/NUCB2 and Its Binding Site in Mouse Testis and Epididymis

  • Kim, Jin-Hee;Kim, Hyun-Sook;Kim, Su-Min;Yang, Hye-Jin;Cho, Hyun-Hae;Hwang, Sup-Yong;Moon, Chan-Il;Yang, Hyun-Won
    • 한국발생생물학회지:발생과생식
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    • 제15권3호
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    • pp.249-256
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    • 2011
  • Nesfatin-1/NUCB2, which is secreted from the brain, is known to control appetite and energy metabolism. Recent studies have been shown that nesfatin-1/NUCB2 was expressed not only in the brain, but it was also expressed in the gastric organs and adipose tissue. However, little is known about the expression of nesfatin-1/NUCB2 in the male reproductive system. Therefore, we examined whether the nesfatin-1/NUCB2 and its binding site exists in the male reproductive organs. Nesfatin-1/NUCB2 mRNA and protein were detected in the mouse testis and epididymis by PCR and Western blot analysis. As a result of the immunohistochemistry staining, the nesfatin-1 protein was localized at the interstitial cells and Leydig cells in the testis. Nesfatin-1 binding sites were also displayed at boundary cells in the tunica albuginea. Furthermore, in order to examine if the expression of nesfatin-1/NUCB2 mRNA in the testis and epididymis were affected by gonadotropin, its mRNA expression was analyzed after PMSG administration into mice. NUCB2 mRNA expression levels were increased in both of the testis and epididymis after PMSG administration. These results demonstrated for the first time that nesfatin-1 and its binding site were expressed in the mouse testis and epididymis. In addition, nesfatin-1/NUCB2 mRNA expression was controlled by gonadotropin, suggesting a possible role of nesfatin-1 in the male reproductive organs as a local regulator. Due to this, further study is needed to elucidate the functions of nesfatin-1 on the male reproductive system.

도시 내 육상 생물종 모니터링을 위한 환경DNA 리뷰 및 적용 (Review and application of environmental DNA (eDNA) investigation of terrestrial species in urban ecosystem)

  • 김휘문;김성열;박일수;이현정;김경태;김영;김혜정;곽민호;임태양;박찬;송원경
    • 한국환경복원기술학회지
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    • 제23권2호
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    • pp.69-89
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    • 2020
  • Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.

한국인의 후두암 발생에서 음주, Aldehyde Dehydrogenase 2(ALDH2)와 N-Acetyltransferase 2(NAT2) 유전자 다형성의 역할 (Effects of Alcohol Intake, Genotypes of Aldehyde Dehydrogenase 2 and N-Acetyltransferase 2 on the Development of Laryngeal Cancer in Koreans)

  • 권순욱;심윤상;이용식;홍성출;김광일;홍영준;홍석일;김현주;김헌;이국행
    • 대한두경부종양학회지
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    • 제17권2호
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    • pp.131-138
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    • 2001
  • Objectives: Alcohol intake has been reported to be a risk factor of laryngeal cancer. Since the aldehyde dehydrogenase 2 (ALDH2) genotype is a major determinant of personal alcohol drinking habit, there is a possibility that ALDH2 genotype would be a risk factor for laryngeal cancer. N-Acetyltransferase 2 (NAT2) is a detoxifying enzyme and its polymorphism has been reported to be related to the risk of many environmental cancers. However, studies on the associations between these two genotypes and laryngeal cancer risk are scarce. We have assessed the effects of alcohol intake and the genotype of ALDH2 and NAT2 on the risk of laryngeal cancer in Koreans. Materials and Methods: Eighty-four pathologically proven laryngeal cancer patients and 168 age matched controls were included as the study subjects. Information about alcohol intake and smoking habit was collected using a self administered questionnaire. ALDH2 and NAT2 genotypes were analyzed using PCR-RFLP methods. Results: Alcohol intake was significant as a risk factor for laryngeal cancer (OR : 2.58, 95% CI : 1.24, 5.36), especially for supraglottic laryngeal cancer (OR : 3.24, 95% CI : 1.02, 10.31). Personal drinking habit was closely related with personal smoking habit, which was a potent risk factor of laryngeal cancer. In a stratified analysis according to the level of cumulative smoking amount, drinking was significant neither in light smokers (equal or less than 30 pack-years) nor in heavy smoker (over 30 pack-years). The ALDH2 genotype was significantly associated with the risk of laryngeal cancer in a univariate analysis. The statistical significance, however, disappeared after adjusting alcohol intake using a multiple conditional logistic model. The NAT2 genotype was not significant as a risk factor for laryngeal cancer. Conclusion: Alcohol drinking and ALDH2 genotype would have indirect effects on laryngeal cancer by their correlations with cigarette smoking or with alcohol drinking. It is less likely that the NAT2 genotype would be a potent risk factor of laryngeal cancer.

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한국인 신생아 황달과 Glutathione S-transferase 다형성에 관한 연구 (Glutathione S-transferase polymorphism of neonatal hyperbilirubinemia in Korean neonates)

  • 강창석;홍승수;김지숙;김은령
    • Clinical and Experimental Pediatrics
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    • 제51권3호
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    • pp.262-266
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    • 2008
  • 목 적 : GSTs는 glutathione과 친전자성 화합물의 결합을 촉매하여 생체내에 독성 물질로부터 조직을 보호하는 효소로, 여러 다형성이 확인 되었으며 일부 GSTs의 null 유전자형을 가진 사람은 GSTs 단백을 생성하지 못하여 다양한 질병의 감수성에 영향을 미친다고 보고 되었다. 이것은 빌리루빈과 같은 non-substrate ligand와 결합하여 세포내로 운반하는 역할을 하는 대표적인 ligandin이며 빌리루빈을 간세포 내 소포체로 이동시켜 UGT를 통해 glucuronidation 시키는 역할을 한다. 이 연구에서는 빌리루빈 대사의 ligandin인 GSTs 중 GSTM1, GSTT1과 신생아 황달과 연관성이 있는 지 알아보고자 본 연구를 시행하였다. 방 법 : 혈청 빌리루빈 수치가 12 mg/dL 이상인 건강하고 위험인자가 없는 만삭아 중 신생아 고빌리루빈혈증 환아 88명, 대조군은 186명을 대상으로 혈액 0.5 cc를 채혈하여 DNA를 분류하였고 중합효소 연쇄 반응을 수행하여 DNA band를 확인하였다. 결 과 : 대조군의 GSTM1 null 유전형 58.1%, GSTT1의 null 유전형 53.2%였다. 환자군에서 GSTM1 null 유전형은 42% (P=0.0187), GSTT1 null 유전형은 31.8% (P=0.0014)로 통계학적 연관성이 있었다. GSTM1/GSTT1 null/null인 경우, 환자군에서 20명(22.7%)(P=0.0008), GSTM1/GSTT1 null/present인 경우 환자군에서 17명(19.3%) (P=0.0470), GSTM1/GSTT1 present/null인 경우 환자군에서 8명(9.1%) (P=0.0066)으로 나타났다 결 론 : GSTM1과 GSTT1 모두 환자군에서 null 유전형이 대조군에 비하여 더 적게 나타나 GSTs null 유전형이 신생아 고빌리루빈혈증의 위험인자는 아니었다.

녹차 카테킨, Epigallocathechin Gallate (EGCG)의 흰쥐췌장종양 선 세포 AR42J의 MAP Kinase 세포 신호전달 기전을 통한 Neurogenin 3 발현에 미치는 영향 (Effect of EGCG on Expression of Neurogenin 3 via the MAP Kinase Signaling Pathway in AR42J Cells, a Rat Pancreatic Tumor Cell Line)

  • 김성옥;최원경
    • Journal of Nutrition and Health
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    • 제44권3호
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    • pp.196-202
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    • 2011
  • 본 연구는 EGCG의 항 당뇨 활성기전으로 췌장종양 선세포 AR42J의 분화 및 내분비기능 개선에 미치는 영향과 그 세포 신호전달 기전을 확인하였다. 그 결과 첫째, AR42J 세포에 EGCG 처리 시 췌장종양 선세포의 세포증식이 농도 의존적으로 감소되었다. 둘째, 세포사멸 유도가 유의적으로 일어나지 않는 농도인 1uM EGCG를 AR42J 세포에 처리한 결과 ngn 3, ${\alpha}$-amylase, insulin은 EGCG처리 24시간에 mRNA, 단백질 발현증가를 나타내었고 48시간에 유의적 증가를 나타내었다. 셋째, EGCG 처리 시 ERK, JNK MAP Kinase 기전은 인산화 억제를 나타내었고 반면에 p38 기전의 인산화는 48시간에 유의적 증가를 하였다. 넷째, p38기전 저해제인 SB203580을 처리하여 EGCG가 MAP Kinase 기전중의 하나인 p38 기전 인산화 활성의 회복을 나타내어 ngn 3 발현을 위한 전사 신호전달 기전임을 다시 확인하였다. 따라서 녹차 생리활성 성분인 EGCG의 췌장종양 선 세포 AR42J 처리 결과 EGCG는 p38 MAP Kinase 기전 활성을 통해 췌장 선세포의 분화지표인 ngn 3 발현을 증가시키며 췌장내분비 기능 지표인 ${\alpha}$-amylase, insulin 발현증가를 나타내어 세포의 내분비기능 개선에도 영향을 미치는 것으로 사료된다.

Diversity of the Gastric Microbiota in Thoroughbred Racehorses Having Gastric Ulcer

  • Dong, Hee-Jin;Ho, Hungwui;Hwang, Hyeshin;Kim, Yongbaek;Han, Janet;Lee, Inhyung;Cho, Seongbeom
    • Journal of Microbiology and Biotechnology
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    • 제26권4호
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    • pp.763-774
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    • 2016
  • Equine gastric ulcer syndrome is one of the most frequently reported diseases in thoroughbred racehorses. Although several risk factors for the development of gastric ulcers have been widely studied, investigation of microbiological factors has been limited. In this study, the presence of Helicobacter spp. and the gastric microbial communities of thoroughbred racehorses having mild to severe gastric ulcers were investigated. Although Helicobacter spp. were not detected using culture and PCR techniques from 52 gastric biopsies and 52 fecal samples, the genomic sequences of H. pylori and H. ganmani were detected using nextgeneration sequencing techniques from 2 out of 10 representative gastric samples. The gastric microbiota of horses was mainly composed of Firmicutes (50.0%), Proteobacteria (18.7%), Bacteroidetes (14.4%), and Actinobacteria (9.7%), but the proportion of each phylum varied among samples. There was no major difference in microbial composition among samples having mild to severe gastric ulcers. Using phylogenetic analysis, three distinct clusters were observed, and one cluster differed from the other two clusters in the frequency of feeding, amount of water consumption, and type of bedding. To the best of our knowledge, this is the first study to investigate the gastric microbiota of thoroughbred racehorses having gastric ulcer and to evaluate the microbial diversity in relation to the severity of gastric ulcer and management factors. This study is important for further exploration of the gastric microbiota in racehorses and is ultimately applicable to improving animal and human health.

LABORATORY STUDIES ON MIC OF AISI TYPE 304 STAINLESS STEEL USING BACTERIA ISOLATED FROM A W ASTEWATER TREATMENT SYSTEM

  • Sreekumari, Kurissery R.;Kyozo, Hirotani;Katsuya, Akamatsu;Takashi, Imamichi;Yasushi, Kikuchi
    • 대한용접접합학회:학술대회논문집
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    • 대한용접접합학회 2002년도 Proceedings of the International Welding/Joining Conference-Korea
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    • pp.260-265
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    • 2002
  • Microbiologically influenced Corrosion (MIC) is one of the most deleterious effects of metal microbe interactions. When a fresh metal surface comes in contact with a non-sterile fluid, biofilm formation is ensued. This might result in the initiation of corrosion. The sites and materials where MIC is implicated are versatile. Industries such as shipping, power generation, chemical etc are reported to be affected. The rapid and unexpected failure of AISI type 304 stainless steel was investigated in the laboratory by simulation studies for a period of 4 months. Slime and water samples from the failure site were screened for corrosion causing bacteria. Both aerobic and anaerobic nora were enumerated and identified using PCR techniques. Pseudomonas sp. and Bacillus sp. were the most common aerobic bacteria isolated from the water and slime samples, whilst sulfate reducing bacteria (SRB) were the major anaerobic bacteria. The aerobic bacteria were used for the corrosion experiments in the laboratory. Coupon exposure studies were conducted using a very dilute (0.1%V/V) nutrient broth medium. The coupons after retrieval were observed under a Scanning Electron Microscope (SEM) for the presence of MIC pits. Compared to sterile controls, metal coupons exposed to Pseudomonas sp and Bacillus sp. showed the initiation of severe pitting corrosion. However, amongst these two strains, Psudomonas sp. caused pits in a very short span of 14 days. Towards the end of the experiment, severe pitting was observed in both the cases. The detailed observation of pits showed they vary both in number and shapes. Whilst the coupons exposed to Bacillus sp. showed widely spread scales like pits, those exposed to Pseudomonas sp. showed smaller and circular pits, which had grown in number and size by the end of the experiment. From these results it is inferred that the rapid and unexpected failure of 304 SS might be due to MIC. Pseudonwnas sp. could be considered as the major responsible bacteria that could initiate pits in the metallic structures. As the appearance of pits was different in both the tested strains, it was thought that the mechanisms of pit formation are different. Experiments on these lines are being continued.

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한국인 건선 환자에서의 IL-1B (-511, +3954)와 IL-1RN 유전자의 다양성 조사 (Investigation of IL-1B (-511, +3954) and IL-1RN Gene Polymorphisms in Korean Psoriasis Patients)

  • 김양겸;표철우;김태윤;김태규
    • IMMUNE NETWORK
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    • 제3권3호
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    • pp.242-247
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    • 2003
  • Background: Psoriasis is an inflammatory skin disorder that is characterized by a marked proliferation of keratinocytes, vascular dilation and leukocyte infiltration. Cytokines play important roles in the pathogenesis of inflammatory disorders. An overexpression of proinflammatory cytokines was characterized in psoriasis plaque. Among these cytokines, IL-$1{\beta}$ is major pro-inflammatory cytokine synthesized during the infection and inflammatory process. The IL-1 receptor antagonist (IL-1Ra) competes for the same IL-1 receptor for $IL-1{\alpha}$ and $-1{\beta}$, which prevents activation of the target cells. Three single nucleotide polymorphisms (SNPs) in the IL-$1{\beta}$ gene have been reported at position -31, -511 and +3954. Within the IL-1Ra gene (IL-1RN), there is a variable number of tandem repeats (VNTR) of an 86 bp length in intron 2. These polymorphisms related to cytokine production and associated with various diseases. Methods: We investigated the polymorphisms of IL-1B (promoter -511 and +3954) and IL-1RN on 114 psoriasis patients and 311 healthy normal controls in Korean. We performed PCR-RFLP on single nucleotide polymorphisms (SNPs) of IL-1B (promoter -511 and +3954) and fragment analysis on IL-1RN 86 bp VNTR polymorphism. Results: The frequency of IL-1B $-511^*1$ allele (patients vs. controls; 50.0% vs. 42.3%, RR=1.4) was significantly increased and IL-1B $-511^*2$ allele (patients vs. controls; 50.0% vs. 57.7%, RR=0.7) decreased in psoriasis patients compared to normal controls. We also analyzed the IL-1B -511 polymorphism according to patients' characters (age of onset, sex and family history). The IL-1B -511 alleles were significantly associated in patients with male and family history than health normal controls. There were no significant associations of IL-1B +3954 and IL-1RN polymorphisms with psoriasis patients. Conclusion: These results suggest that the polymorphism of IL-1B -511 could be genetic susceptibility to psoriasis in Koreans.

양측성 중증 폐렴으로 인한 ARDS로 오진된 속립성 결핵 (A Case of Miliary Tuberculosis Mimicking ARDS due to Bilateral Severe Pneumonia)

  • 한군희;정복현;김영돈;황정원;홍현일;윤성규;김미혜;류대식;강길현
    • Tuberculosis and Respiratory Diseases
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    • 제59권1호
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    • pp.109-113
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    • 2005
  • 저자들은 중증 폐렴으로 인한 ARDS양상으로 내원한 임신 3개월의 속립성 결핵 환자를 경기관지 폐생검과 방사선 소견 및 임상양상을 종합적으로 고려하여 속립성 결핵에 의한 ARDS로 확진할 수 있었던 1예를 경험하였기에 문헌 고찰과 함께 보고하는 바이다.

The Differential Staging of Murine Thymic Lymphoma Cell Lines, Scid.adh, R1.1 and EL-4

  • Chae, Jong Seok;Kim, Hae-jung;Park, Weon Seo;Bae, Youngmee;Jung, Kyeong Cheon
    • IMMUNE NETWORK
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    • 제2권4호
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    • pp.217-222
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    • 2002
  • Background: Scid.adh is a recently developed murine thymic lymphoma cell line, which has been used as in vitro model for the study of double negative stage III thymocytes. In this study, we compared the expression profile of a number of genes and proteins, which are tightly related to T cell development and apoptosis, in thymic lymphoma cell lines, R1.1, EL-4, and Scid.adh for the developmental staging. Methods: We examined the expression of development marker genes and proteins in three lymphoma cell lines by flow cytometry and RT-PCR. In addition, the expression of apoptosis-related molecules including bcl-2, bax and Fas was also investigated. Results: As previously reported, Scid.adh cell line expressed CD8 and CD25 but not TCR ${\alpha}$ chain, while R1.1 cells expressed TCR ${\alpha}$ chain and both CD4 and CD8 transcripts. These suggest that R1.1 might be in double positive stage, and low level of CD44 expression and the absence of CD25 support this suggestion. In contrast, EL-4 cells showed high level of TCR ${\alpha}$ chain transcript, and low-level of CD4 expression, suggesting that EL-4 is in more mature stage than R1.1. Further, this suggestion was supported by the lack of mT-20 in EL-4 cells, which is expressed in the immature thymocytes, and Scid.adh and R1.1 cell lines, but not in the terminally differentiated thymocytes and peripheral T cells. Among the apoptosis-related gene, transcripts of bcl-2 gene were detected in both R1.1 and EL-4 but not in Scid.adh cells, while bax was expressed in all cell lines. Fas expression was the highest in EL-4 cells and low in Scid.adh cell line. Conclusion: R1.1 cell may represent double positive stage, and EL-4 is more differentiated cell line. In addition, Scid.adh and EL-4 cell lines are suspected to be useful for the study of function of bcl-2 family and Fas during the thymocyte development, respectively.