• Parasites, Hosts and Diseases
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• 제30권1호
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• pp.33-42
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• 1992

자유생활 아메바인 Naegleria fowleri는 비강을 통해 인체에 들어와 출혈성 수막뇌염을 일으켜 감염된 사람의 대부분을 일주일 이내에 사망하게 하는 원충이다. 이 실험에서는 N. fowleri에 특이한 단세포군 항체를 만들어 이의 특성 및 그 이용 가능성을 알아보고자 하였다. 7 종류의 단세포군 항체(Nf 1, Nf 2, NE 256, Nf 279, Nf 27, Nf 154, Nf 137)를 제조하였는데 각각의 isotope은 IgGl이 두 종류(Nf 27, Nf 154), IgG3가 1 종류(Nf 137), IgA가 4 종류(Nf 1, Nf 2, Nf 256, Nf 279)이었다. 이들 항체를 N. fwoleri와 다른 종류의 아메바를 항원으로 하여 효소표식 면역검사법을 시행하여 Nf 1 및 Nf 256 항체를 제외한 5종류의 항체가 N. fowleri에 특이한 항체임을 확인하였다. 또한 간접 형광항체법을 통하여 NF 256 항체를 제외한 6 종류의 단세포군 항체가 세포막의 일부에 결합하였음을 관찰하였다. Nf 154 항체를 이용한 immunoperoxidase 염색에서도 세포막의 반응을 관찰하였다. 단세포를 항체가 N. fowleri를 응집시키는지 알아보고자 항체를 N. fowleri 영양형과 반응시켰더니 Nf 27 항 체를 제외한 5 종류의 단세포를 항체에 의해 응집이 일어남을 알 수 있었으며 응집된 덩 양형을 보체로 처리한 후 CGVS배지에서 배양하였더니 영양형의 증식이 억제되었다. 또한 2종류의 단세포군 항체(Nf 2, Nf 154)는 조직 세포(Chinese hamster ovary cell: CHO)에 대한 N.fowleri의 세포독성을 저하시켰다. 단세포군 항채와 반응하는 항원의 분자량을 알아보고자 EITB(Enzyme-linked immunoelectrotransfer blot)를 시행하였는데 Nf 279 항체는 25 kDa 및 28 kDa, Nf 154 항체는 43 kDa, N( 137 항체는 29 kDa에 해당되는 분획에서 항원항체반응을 나타내었다. 이상의 성적을 종합하면 N. fowleri에 대한 7종류의 IgG., IgG3 및 IgA 단세포군 항체를 생산하였다. 그중 Nf 256 항체를 제외한 6 종류는 N. fowleri이 세포막 성분중 28 kDa-43 kDa의 항원과 반응하는 특이한 항체임을 관찰할 수 있었다. 또한 이 단세포를 항체들은 영양형을 응집시키며, 시험관 내에서의 증식을 억제시키고 CHO 세포에 대한 N. fowleri의 세포 독성을 저하시키는 성질을 가지고 있음을 알 수 있었다.

• BMB Reports
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• 제33권6호
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• pp.519-524
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• 2000

NF1 proteins are a family of DNA binding proteins which consist of two separate domains, N-terminal DNA binding domain and C-terminal transcription activation domain. The N-terminal 220 amino acids are highly conserved and are also known to mediate dimerization of NF1 proteins. The C-terminal regions of different type of NF1 proteins are heterogeneous and responsible for transcriptional activation. In this study, we tested the interaction between different domains of rat NF1-A protein by yeast two hybrid analysis and observed the interaction between C-terminal regions of NF1-A which do not contain the N-terminal dimerization domain. Our results showed that the C-terminal region of rat NF1-A between residues 231 and 509 strongly interacted not only with itself, but also with human NF1/CTF1 which is a different type of NF1. When the C-terminal region was divided into two fragments, one from residue 231 to 447 and the other from 448 to 509, the two fragments were able to interact with the C-terminal region of NF1-A significantly. This indicates that both fragments contain independent interaction domains. Analysis of the interactions with alanine substituted fragments showed that substitutions of the heptasequence, SPTSPTY of NF1-A, affected interaction between NF1 proteins. Our results strongly suggest that C-terminal regions may also be important for the formation of homo- and heterodimers in addition to the N-terminal dimerization domain. Also, the heptasequence motif may play some roles in dimer formation.

• BMB Reports
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• 제44권8호
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• pp.553-557
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• 2011

We previously reported that CDK2/Cyclin A can phosphorylate and activate the transcription factor NF-Y. In this study, we investigated a potential regulatory role for NF-Y in the transcription of Cyclin A and other cell cycle regulatory genes. Gel-shift assays demonstrate that NF-Y binds to CCAAT sequences in the Cyclin A promoter, as well as to those in the promoters of cell cycle G2 regulators such as CDC2, Cyclin B and CDC25C. Furthermore, expression of Cyclin A increases NF-Y's affinity for CCAAT sequences in the CDC2 promoter; however, Cyclin A's induction of CDC2 transcription is antagonized by p21, an inhibitor of CDK2/Cyclin A. These results suggest a model wherein NF-Y binds to and activates transcription from the Cyclin A promoter, increasing cellular levels of Cyclin A/CDK2 and potentiating NF-Y's capacity for transcriptional transactivation, and imply a positive feedback loop between NF-Y and Cyclin A/CDK2. Our findings are additionally indicative of a role for Cyclin A in activating Cyclin B/CDK1 through promoting NF-Y dependent transcription of Cyclin B and CDC2; NF-Y mediated crosstalk may therefore help to orchestrate cell-cycle progression.

• Journal of Genetic Medicine
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• 제21권1호
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• pp.22-30
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• 2024

Purpose: Neurofibromatosis 1 (NF1) is one of the most common autosomal dominant diseases caused by heterozygous mutation in the NF1 gene. Mutation detection is complex owing to the large size of the NF1 gene, the presence of a high number of partial pseudogenes, and the great variety of mutations. We aimed to study the mutation spectrum of NF1 gene in Korean patients with NF1. Materials and Methods: We have analyzed total 69 unrelated patients who were clinically diagnosed with NF1. PCR and sequencing of the NF1 gene was performed in all unrelated index patients. Additionally, multiplex ligation-dependent probe amplification (MLPA) test of the NF1 and SPRED1 gene analysis (sequencing and MLPA test) were performed in patients with negative results from NF1 gene sequencing analysis. Results: Fifty-five different variants were identified in 60 individuals, including six novel variants. The mutations included 36 single base substitutions (15 missense and 21 nonsense), eight splicing mutations, 13 small insertion or deletions, and three gross deletions. Most pathogenic variants were unique. The mutations were evenly distributed across exon one through 58 of NF1, and no mutational hot spots were found. When fulfilling the National Institutes of Health criterion for the clinical diagnosis of NF1, the detection rate was 84.1%. Cafe-au-lait macules were observed in all patients with NF1 mutations. There is no clear relationship between specific mutations and clinical features. Conclusion: This study revealed a wide spectrum and genetic basis of patients with NF1 in Korea. Our results aim to contribute genetic management and counseling.

• 대한한방소아과학회지
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• 제15권2호
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• pp.69-73
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• 2001

The neurofibromatosis (NF) are a set of genetic disorders which cause tumors to grow along various types of nerves and, in addition, can affect the development of non-nervous tissues such as bones and skin. NF causes tumors to grow anywhere on or in the body. It also leads to developmental abnormalities. For example, individuals with NF have a higher incidence of learning disabilities. Neurofibromatosis(NF) has been classified into two distinct types: NF-I and NF-II. neurofibromatosis 1(NF-I), also known as von Recklinghausen NF or Peripheral NF, occurring in 1:4,000 births, is characterized by multiple cafe-au-lait spots and neurofibromas on or under the skin. Enlargement and deformation of bones and curvature of the spine (scoliosis) may also occur. Occasionally, tumors may develop in the brain, on cranial nerves, or on the spinal cord. About 50% of people with NF also have learning disabilities. Neurofibromatosis 2(NF-II), also known as Bilateral Acoustic NF(BAN), is much rarer occurring in 1:50,000 births. NF-II is characterized by multiple tumors on the cranial and spinal nerves, and by other lesions of the brain and spinal cord. Tumors affecting both of the auditory nerves are the hallmark. Hearing loss beginning in the teens or early twenties is generally the first symptom. We reported a 10-year-old female patient with NF-I, she has pain and edema in left leg, no symptoms of NF.

• Molecules and Cells
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• 제20권2호
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• pp.241-246
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• 2005

PDK-1 activates PI3-kinase/Akt signaling and regulates fundamental cellular functions, such as growth and survival. NF-${\kappa}B$ is involved in the induction of a variety of cellular genes affecting immunity, inflammation and the resistance to apoptosis induced by some anti-cancer drugs. Even though the crucial involvement of the PI3-kinase/Akt pathway in the anti-apoptotic activation of NF-${\kappa}B$ is well known, the exact role of PDK-1 as well as PI3-kinase/Akt in NF-vactivation is not understood. Here we demonstrate that PDK-1 plays a pivotal role in transcriptional activation of NF-${\kappa}B$ by dissociating the transcriptional co-repressor HDAC1 from the p65 subunit of NF-${\kappa}B$. The association of CBP with p65 was not directly modulated by PDK-1 or by PI3-kinase. Etoposide activated NF-${\kappa}B$ through PI3-kinase/Akt, and the transcription activation domain (TAD) of p65 was further activated by wild-type PDK-1. Overexpression of a dominant negative PDK-1 mutant decreased etoposide-induced NF-${\kappa}B$ transcription and further down-regulated the ectopic HDAC1-mediated decrease in NF-${\kappa}B$ transcriptional activity. Thus activation of PDK-1 relieves the HDAC1-mediated repression of NF-${\kappa}B$ that may be related to basal as well as activated transcription by NF-${\kappa}B$. This effect may also explain the role of the PI3-kinase/PDK-1 pathway in the anti-apoptotic function of NF-${\kappa}B$ associated with the chemoresistance of cancer cells.

• Molecules and Cells
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• 제20권1호
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• pp.105-111
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• 2005

Mannasantin B, a dilignan structurally related to manssantin A, is an inhibitor of NF-${\kappa}B$ transactivation. In the present study, we found that it inhibited PMA-induced expression of IL-$1{\beta}$, IL-$1{\beta}$ mRNA, and IL-$1{\beta}$ promoter activity in U937 cells with $IC_{50}$ values of about 50 nM. It also inhibited NF-IL6- and NF-${\kappa}B$-induced activation of IL-$1{\beta}$, with $IC_{50}$ values of 78 nM and $1.6{\mu}M$, respectively, revealing a potent inhibitory effect on NF-IL6. Electrophoretic mobility shift assays showed that manassantin B had an inhibitory effect on DNA binding by NF-IL6, but not by NF-${\kappa}B$. Further analysis revealed that transactivation by NF-IL6 was also inhibited. Our results indicate that manassantin B suppresses expression of IL-$1{\beta}$ in promonocytic cells by inhibiting not only NF-${\kappa}B$ but also NF-IL6 activity. Furthermore, our observations suggest that manassantin B may be clinically useful as a potent inhibitor of NF-IL6 activity.

• 한국지구과학회지
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• 제39권6호
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• pp.519-532
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• 2018

우리는 오리온 분자운 복합체의 북부 필라멘트(이하 NF)에 대하여 $^{12}CO$ (J=1-0) 분자선의 자료를 이용하여 은하 평면이 분자의 운동과 운동학에 미치는 영향을 연구하였다. 6 m 서울대학교 전파망원경(Seoul Radio Astronomy Observatory, SRAO)을 이용하여 2arcmin 공간분해능으로 은하면으로부터 먼 순서로 NF1, NF2, NF3 세 곳을 총 270시간 동안 관측된 자료를 사용하였다. 은하면과 OMC NF는 $^{12}CO$ (J=2-1) 경우 3% 밀도에서, 티끌의 경우 9% 밝기 수준에서 자기장을 따라 서로 연결되어 있었다. $^{12}CO$ (J=1-0), $^{12}CO$ (J=2-1), 성간 티끌 관측결과를 비교해본 결과, 세 경우 모두 NF3에서는 고루 분포했지만, NF1과 NF2에서는 비교적 밀도가 높은 특정 영역에서만 함께 나타났다. NF는 단일 구조를 보였으며, NF1에서는 부분 수축 운동을, NF2에서는 하단에서 회전 운동이 나타났고, NF3에서는 유일하게 명확히 자기장에 연관된 나선형 회전이 보였다. 위치-속도 분석 결과, $^{12}CO$ (J=1-0)를 비롯한 물질들은 NF2와 NF3을 따라 은하면을 향하여 흐를 가능성이 있음을 확인할 수 있었다. 은하면을 향하여 물질이 흐르는 명백한 원인을 이번 연구결과에서 볼 수 없었지만 추후의 더 정교한 관측결과가 NF1과 NF2 상단부의 회전 운동을 확인 할 수 있겠다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.128-128
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• 2018

Nypa fruticans wurmb (NF) has been used as traditional medicinal food in Asian countries. Especially, NF has been used for conventional medicine to treat inflammatory periodontal diseases. Previous studies have been shown that NF has large amount of useful constituents such as phenolic acids, polyphenols and flavonoids. Also, NF is known as having medicinal effects such as anti-oxidant, anti-inflammatory and cholesterol-lowering effects. NF has recently been attracted to use complementary medicinal food on inflammatory diseases in Korea. However, there are no obvious effects in inflammatory and metabolic diseases also mechanisms has been studied yet. The purpose of this study was to investigate the anti-inflammatory effects of NF and steamed-NF (SNF), which recently has been used as health food, using Human keratinocyte cell line (HaCaT) and Human mast cell line (HMC-1). The cytotoxicities of NF and SNF were measured by using MTT assays in HaCaT cells and HMC-1 cell. To evaluate anti-inflammatory effects of NF and SNF, HaCaT cells were stimulated with tumor necrosis factor $(TNF)-{\alpha}$ and Interferon $(IFN)-{\gamma}$. Also, HMC-1 cells were stimulated with phorbol-12-myristate-13-acetate (PMA) and A23187 calcium ionophore (A23187) to induce allergic inflammation. Inflammatory cytokine were measured by enzyme-linked immunosorbent assay (ELISA). In this result, the extract of NF and SNF (0.01 - 1mg/ml) did not show cytotoxicity in HaCaT cells and HMC-1 cells. In addition, the NF and SNF suppressed the production of interleukin (IL)-6 and IL-8 in HaCaT cells at highest concentration. Furthermore, the treatment of SNF significantly inhibited the secretion level of IL-8 in PMA plus A23187-stimulated HMC-1 cells compared with NF treatment group. These results suggest that the extract of NF and SNF may serve as a potential therapy for skin inflammatory diseases.

• Tuberculosis and Respiratory Diseases
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• 제55권5호
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• pp.488-498
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• 2003

연구배경 : 세포에 방사선을 조사하면 세포사멸과 함께 AP-l, NF-${\kappa}B$와 같은 여러 전사인자가 활성화 되는 것으로 알려져 있다. 이중 염증과 면역반응의 중요한 전사인자인 NF-${\kappa}B$는 항아포프토시스의 기능이 있으며 NF-${\kappa}B$ 활성화를 차단함으로써 TNF-${\alpha}$나 daunorubicine 등에 의 한 항암효과를 상승시킬 수 있음이 밝혀져 있다. NF-${\kappa}B$의 항아포프토시스 기전은 NF-${\kappa}B$ 의존성 단백질인 cIAPI, 2의 전사발현 유도에 의한 것으로 cIAPl, 2는 caspase 3, 7, pro-caspase-9의 활성을 차단함으로써 아포프토시스를 억제하는 것으로 알려져 있다. 이에 저자들은 방사선 유도성 세포사멸에 비교적 내성을 보이는 A549 세포주에서 방사선에 의한 NF-${\kappa}B$ 활성화와 그에 따른 cIAP 발현유도를 조사하고 NF-${\kappa}B$ 활성화를 차단하여 방사선 유도성 세포사멸의 감작효과를 확인하기 위하여 본 연구를 시행하였다. 방 법 : 세포주는 A549 폐암세포주를 이용하였고, 방사선 조사는 Varian사의 Clinac 1800C 선형가속기를 이용하였으며 조사량은 10GY를 사용하였다. 세포독성 검사는 MTT Assay를 이용하였고, NF-${\kappa}B$ 활성화 검사는 luciferase reporter gene assay, electromobility shift assay, $I{\kappa}B-{\alpha}$ degradation에 대한 westem blot을 이용하였다. NF-${\kappa}B$활성을 차단하기 위하여 proteosome inhibitor인 MGI32와 $I{\kappa}B{\alpha}$-superrepressor plasmid를 transfection한 안정적 세포주 A549-$I{\kappa}B{\alpha}$-superrepressor를 이용하였다. cIAP의 발현은 RT-PCR을 이용하였고, cIAP2 promoter 활성은 NF-${\kappa}B$ site를 포함한 cIAP2 유전자 5' f1anking region(1.4kb)을 pGL2-Basic luciferase vector에 cloning 한 construct를 사용하여 transfection 후 luciferase assay를 시행하였다. 결 과 : A549 cell에서 10Gy 방사선 조사에 의한 세포독성은 24hr, 48hr에 각각 $10.82{\pm}.3%$, $17.7{\pm}6.4%$로 비교적 내성이 있음을 확인하였다. 방사선에 의한 NF-${\kappa}B$의 활성은 $I{\kappa}B-{\alpha}$ 분해에 대한 western blot과 EMSA로 확인하였으며 luciferase assay에서도 약 1.6 배 정도의 NF-${\kappa}B$ 활성화가 있었다. NF-${\kappa}B$활성을 차단하기 위해 사용한 MG132는 방사선 유도성 세포사멸에 영향을 주지 않았으며 A549-$I{\kappa}B{\alpha}$-superrepressor 세포주에서도 세포사멸의 감작효과는 없었다. 또한 RT-PCR 결과 방사선에 의한 cIAP1,2 mRNA 발현유도는 관찰되지 않았고 cIAP2 promoter luciferase assay에서도 cIAP2 전사활성 유도는 없었다. 결 론 : A549 폐암세포주에서 방사선에 의해 NF-${\kappa}B$의 활성화는 확인하였으나 활성화 정도가 미약하였고 NF-${\kappa}B$ 의존성 항아포프토시스 유전자인 cIAP가 방사선에 의해 발현유도 되지 않았으며, NF-${\kappa}B$ 활성을 차단함에도 세포독성에 감작효과가 없었으므로 A549 폐암세포주에서 방사선 유도성 세포사멸에 내성을 보이는 기전에는 NF-${\kappa}B$의 역할이 미미하리라고 사료된다.