• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.658-671
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• 2018

Objective: Nandrolone decanoate (ND) is an anabolic-androgenic steroid frequently used for clinical treatment. However, the inappropriate use of ND results in the reduction of serum testosterone level and sperm production. The suppressive effect of ND on testosterone production has not been investigated in detail. The present study was designed to examine the effect of ND on the expression of steroidogenic enzymes in the rat testis. Methods: Male Sprague Dawley rats at 50 days of age were subcutaneously administrated with either 2 or 10 mg of ND/kg body weight/week for 2 or 12 weeks. The changes of transcript and protein levels of steroidogenic enzymes in the testis were determined by real-time polymerase chain reaction and western blotting analyses, respectively. Moreover, immunohistochemical analysis was employed to determine the changes of immunostaining intensity of these enzymes. The steroidogenic enzymes investigated were steroidogenic acute regulatory protein, cytochrome P450 side chain cleavage enzyme, $17{\alpha}-hydroxylase$, $3{\beta}-hydroxysteroid$ dehydrogenase, and cytochrome P450 aromatase. Results: The treatment of ND resulted in depletion of Leydig cells and sloughing of germ cells in the testis. The ND treatment caused significant expressional decreases of steroidogenic enzymes at transcript and protein levels, and the destructive effects of ND on the testis were more apparent with a higher dose and a longer period of the treatment. Evident reduction of immunostaining intensity present in Leydig cells was clearly detected by the ND treatment. Conclusion: The exposure to ND in young male results not only in histological changes of the testis but also in aberrant gene expression of testicular steroidogenic enzymes, consequently leading into the reduction of testosterone production in the testis and thus likely disruption of spermatogenesis.

• 동의생리병리학회지
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• 제19권1호
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• pp.81-86
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• 2005

The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Psoraleae fructus. The extract was studied for dipheny-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being 0.427 mg/mL. The extract was dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (67.7 %) was blocked by the extract concentration- dependently. Furthermore, the extract also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Psoraleae fructus has potent antioxidant activity.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1267-1274
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• 2004

Colostrum contains various kinds of cytokines including TGF-$\beta$ that has potent regulatory effects on cells of the immune system. We compared the levels of TGF-$\beta$1 and TGF-$\beta$2 in bovine and human colostrum. Based on the isoform-specific ELISA, bovine colostrum collected on day 1 post-delivery retained $53.71{\pm}29.55\;ng/ml$ of TGF-$\beta$1 and $40.41{\pm}21.78\;{\mu}g/ml$ of TGF-$\beta$2 (n=4), while in human, $381.45{\pm}158.24\;ng/ml$ of TGF-$\beta$1 and $41.47{\pm}9.63\;ng/ml$ of TGF-$\beta$2 (n=5). Thus, dominant TGF-$\beta$ isoforms were completely opposite between human and bovine colostrum samples. The concentrations of both isoforms declined as lactation proceeded. Biological activities of the colostrum samples were determined using an MV1LU cell line. Consistent with the result from the immunoassay, TGF-$\beta$1 in human and TGF-$\beta$2 in bovine colostrum were responsible for the anti proliferative activity against MV1LU cells. Furthermore, bovine colostrum increased IgA secretion by LPS-stimulated mesenteric lymph node (MLN) cells, and this effect was abrogated by either anti­TGF-$\beta$2 antibody or combined anti-TGF-$\beta$1/$\beta$2 antibody, but not by anti- TGF-$\beta$1 antibody alone. Similarly, TGF-$\beta$2 in bovine colostrum enhanced the Ig germ line (GL) promoter activity, which is the earliest event toward IgA isotype switching. Taken together, these results suggest that TGF-$\beta$ isoforms, differentially expressed in human and bovine colostrum, may promote IgA isotype production in the neonatal intestine.

• Asian-Australasian Journal of Animal Sciences
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• 제28권11호
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• pp.1565-1572
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• 2015

Porcine embryonic stem cells (pESCs) have become an advantageous experimental tool for developing therapeutic applications and producing transgenic animals. However, despite numerous reports of putative pESC lines, deriving validated pESC lines from embryos produced in vitro remains difficult. Here, we report that embryo aggregation was useful for deriving pESCs from in vitro-produced embryos. Blastocysts derived from embryo aggregation formed a larger number of colonies and maintained cell culture stability. Our derived cell lines demonstrated expression of pluripotent markers (alkaline phosphatase, Oct4, Sox2, and Nanog), an ability to form embryoid bodies, and the capacity to differentiate into the three germ layers. A cytogenetic analysis of these cells revealed that all lines derived from aggregated blastocysts had normal female and male karyotypes. These results demonstrate that embryo aggregation could be a useful technique to improve the efficiency of deriving ESCs from in vitro-fertilized pig embryos, studying early development, and deriving pluripotent ESCs in vitro in other mammals.

• 한국식물병리학회지
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• 제2권1호
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• pp.37-42
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• 1986

[ $0.5\%$ ] lactophenol acid fuchsin으로 염색해 본 결과 보리흰가루 병균의 제1차 발아관에 반응하여 보리표피 세포에 papillae 및 cytoplasmic aggregate가 형성되었으나 그 크기는 부착기에 반응하여 형성된 것보다는 훨씬 작았으며 또한 접종후 36-48시간 이후의 표피세포내에는 acid fuchsin에 의해서 다른 세포들에 비해 세포전체가 좀 더 진하게 염색되는 곳이 군데군데 관찰되었다. 그러나 접종 후 96시간까지 지질과산화산물의 일종인 malondialdehyde의 함량은 증가하지 않았다. 접종 후 6시간에 형성된 papillae 및 cytoplasmic aggregate내에 callose, protein, phenol물질 등은 집적되었으나 접종 후 72시간까지도 cellulose, cutin, suberin, lignin등은 검출되지 않았다. 보리-흰가루병 상호조합의 상기 모든 반응은 race 비특이적이었다.

• 한국식물병리학회지
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• 제3권2호
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• pp.100-107
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• 1987

동일한 벼 품종은 수분함량이 다른 토양에서 재배했을 경우, 도열병에 대한 감수성에 차이가 생기는데 그 원인을 조사하기 위하여 잎 표피에서 분생포자발아 및 부착기형성까지의 도열병균 침입전 행동을 광학현미경과 주사전자현미경(SEM)으로 관찰하였다. 수분함량이 다른 토양에서 자란 벼 잎 사이에 표피세포의 외부형태, 잎 표피에서의 분생포자발아관의 생장 및 생장방향, 부착기의 형태 및 크기는 차이가 없었다. 부착기는 토양수분처리와 상관없이 기동세포$(35\~48\%)$위에 가　 많이 형성되었고 단세포$(19\~27\%)$, 장세포 및 공변세포$(13\~20\%)$의 순이었다. 모분상의 부착기형성은 관찰되지 않았다. 본 시험결과, 수분함량이 다른 토양에서 자란 벼 사이에 나타나는 도열병에 대한 감수성의 차이는 도열병균의 침입전 행동에서 기인하는 것이 아닌 것으로 생각된다.

• Biomolecules & Therapeutics
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• 제22권5호
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• pp.371-383
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• 2014

The nematode Caenorhabditis elegans (C. elegans) offers a unique opportunity for biological and basic medical researches due to its genetic tractability and well-defined developmental lineage. It also provides an exceptional model for genetic, molecular, and cellular analysis of human disease-related genes. Recently, C. elegans has been used as an ideal model for the identification and functional analysis of drugs (or small-molecules) in vivo. In this review, we describe conserved oncogenic signaling pathways (Wnt, Notch, and Ras) and their potential roles in the development of cancer stem cells. During C. elegans germline development, these signaling pathways regulate multiple cellular processes such as germline stem cell niche specification, germline stem cell maintenance, and germ cell fate specification. Therefore, the aberrant regulations of these signaling pathways can cause either loss of germline stem cells or overproliferation of a specific cell type, resulting in sterility. This sterility phenotype allows us to identify drugs that can modulate the oncogenic signaling pathways directly or indirectly through a high-throughput screening. Current in vivo or in vitro screening methods are largely focused on the specific core signaling components. However, this phenotype-based screening will identify drugs that possibly target upstream or downstream of core signaling pathways as well as exclude toxic effects. Although phenotype-based drug screening is ideal, the identification of drug targets is a major challenge. We here introduce a new technique, called Drug Affinity Responsive Target Stability (DARTS). This innovative method is able to identify the target of the identified drug. Importantly, signaling pathways and their regulators in C. elegans are highly conserved in most vertebrates, including humans. Therefore, C. elegans will provide a great opportunity to identify therapeutic drugs and their targets, as well as to understand mechanisms underlying the formation of cancer.

• 동의생리병리학회지
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• 제19권6호
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• pp.1541-1545
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• 2005

The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Corm fructus. The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being $200{\mu}g/mL$. The extract at concentrations of $10-500{\mu}g/ml$ showed dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (63.0%) was blocked by the extract (10, 50, 100, 250 and $500{\mu}g/ml$) concentration-dependently. Furthermore, the extract (50, 100 and $250{\mu}g/ml$) also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Corm fructus has potent antioxidant activity.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.50-50
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• 2003

We have identified and analysed a putative response regulator two-component gene (CaSKN7) from Candida albicans and its encoding protein (CaSkn7). CaSKN7 has an open reading frame of 1677bp. CaSKN7 encodes a 559 amino acid protein (CaSkn7) with an estimated molecular mass of 61.1 kDa. CaSKN7 is a homologue of a Saccharomyces cerevisiae SKN7 that is the regulator involved in the oxidative stress response. To study the role of CaSKN7, we constructed a CAI4-derived mutant strain carrying a homozygous deletion of the CaSKN7 gene. In the caskn7 disruptant cells, the formation of germ tube require shorter time than that in the congenic wild-type strain but the growth of mycelium delayed in liquid media. In contrast, the caskn7 disruptant cells attenuate the differentiation in solid media and the virulence in mouse model system. Expression level of hypha-specific and virulence genes - HYR1, ECE1, HWP1, and ALS1 - in the caskn7 disruptant cells increased as compared with that in the congenic wild-type strain in 10％ serum YPD. Skn7 in 5. cerevisiae was found to bind the HSE element from the SSA promoter, Also, CaSkn7 contains heat shock factor DNA-binding domain and the promoters of these genes have HSE-like sties. Therefore these results show that CaSKN7 regulate the differentiation and virulence of C. albicans.

• IMMUNE NETWORK
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• 제4권4호
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• pp.216-223
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• 2004

Background: It is well known that IgA isotype switching is induced by $TGF-{\beta}1$. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of $TGF-{\beta}1$. Nevertheless, there are lots of difficulties in studying normal B cells in detail because it is not simple to obtain highly purified B cells, showing low reproducibility and transfection efficacy, moreover impossible to keep continuous culture. To overcome these obstacles, it is desperately needed to develop B cell line which acts like normal B cells. In the present study, we investigated whether CH12F3-2A lymphoma cells are appropriate for studying IgA isotype switching event. Methods: CH12F3-2A B cell line was treated with LPS and $TGF-{\beta}1$, then levels of germ-line (GL) transcripts were measured by RT-PCR, and $GL{\alpha}$ promoter activity was measured by luciferase assay. In addition, membrane IgA (mIgA) expression and IgA secretion were determined by FACS and ELISA, respectively. Results: $TGF-{\beta}1$, regardless of the presence of LPS, increased level of $GL{\alpha}$ transcripts but not $GL{\gamma}2b$ transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and $TGF-{\beta}1$. Both mIgA and IgA secretion in the presence of $TGF-{\beta}1$ were further increased by over-expression of Smad3/4. Finally, $GL{\alpha}$ promoter activity was increased by $TGF-{\beta}1$. Conclusion: CH12F3-2A cell line acts quite similarly to the normal B cells which have been previously reported regarding IgA expression. Thus, CH12F3-2A lymphoma cell line appears to be adequate for the investigation of the mechanism(s) of IgA isotype switching at the cellular and molecular levels.