• Title/Summary/Keyword: Small protein

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The Regulation of FOXP3 Expression by the Treatment of TGF-${\beta}$ and the Modification of DNA Methylation in Lung Cancer Cell Lines

  • Um, Sang-Won;Lee, Sang-Hee;Kim, Ho-Joong;Kwon, O-Jung;Kim, Hang-Rae;Kang, Jae-Seung;Lee, Wang-Jae
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.3
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    • pp.206-217
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    • 2011
  • Background: Transcription factor FOXP3 characterizes the thymically derived regulatory T cells. FOXP3 is expressed by cancer cell itself and FOXP3 expression was induced by TGF-${\beta}$ treatment in pancreatic cancer cell line. However, the expression of FOXP3 expression is not well known in patients with lung cancer. This study was conducted to investigate the expression of FOXP3 in patients with lung cancer and to investigate the regulation of FOXP3 expression by the treatment of TGF-${\beta}$ and DNA methyltransferase inhibitor in lung cancer cell lines. Methods: FOXP3 expression in the tissue of patients with resected non-small cell lung cancer (NSCLC) was evaluated by immunohistochemistry. The regulation of FOXP3 expression was investigated by Western blot and RT-PCR after lung cancer cell lines were stimulated with TGF-${\beta}1$ and TGF-${\beta}2$. The regulation of FOXP3 expression was also investigated by RT-PCR and flow cytometry after lung cancer cell lines were treated with DNA methyltransferase inhibitor (5-AZA-dC). Results: FOXP3 expression was confirmed in 27% of patients with NSCLC. In NCI-H460 cell line, TGF-${\beta}2$ decreased FOXP3 mRNA and protein expressions. In A549 cell line, both TGF-${\beta}1$ and TGF-${\beta}2$ decreased FOXP3 mRNA and protein expressions. 5-AZA-dC increased FOXP3 mRNA expression in NCI-H460 and A549 cell lines. Moreover, 5-AZA-dC increased intracellular FOXP3 protein expression in A549 cell lines. Conclusion: It was shown that FOXP3 is expressed by cancer cell itself in patients with NSCLC. Treatment of TGF-${\beta}2$ and DNA methyltransferase inhibitor seems to be associated with the regulation of FOXP3 expression in lung cancer cell lines.

A Study on the Nutritional Status of Lunch Box of Elementary School Students in Kyungnam Area (경남지역 국민학교 아동의 도시락 영양실태에 대한 조사연구)

  • 이성숙
    • Journal of the Korean Home Economics Association
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    • v.25 no.3
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    • pp.49-61
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    • 1987
  • The main purpose of these studies was the improvemen of nutritional supply and of the future physical coditions. The two groups which were regional difference were chosen and research for the nutritional status of elementary school students. The nutritional status of lunch-box between one elementary School of 98 students in the 6th year grade(hence-forward it was called "A-Group"), located at the center of Masan City, being provided the milk and the other elementary School of 134 students in the 6th year grade(hence-forward it was called "B-Group"), located in the small farm and fishing villages of Kusan-myun, Euichang-kun, Kyungnam province, were seweyed from the period of Nov. 20, 1986 to Dec. 4, 1986 and conclusion were as follows: 1. Nutritional status of lunch-box 1) It is indicated that in the case of A-Group, the nutirtional intake except animal protein and vitamin A were lower than the Recommended Daily Allowances of Korenas and in the case B-Group, was also lower than except vitamin A respectively and especially in both groups, the percentage of Fe was lowest and intended difference between A Group and B was that B-Group in the intake of calorie, fat, Ca, Fe and etc were lower than A-Group. 2) The total calorie-intake was short of the extent to approx. 83.8% as compared to the RDA. On the viewpoint at ratio of intake of caloric nutrients, A-Group appeared to be 69.5 : 16.5 : 14 and B-Group, 77.5 : 19.5 : 13 in order carbohydrate, protein and fat. As a result of the above ratio, B-Group depended upon carbohydrate as a staple food more than A-Group. It can be considered that the total calorie-intake is short, and that the amount of lunch-box was greatly insufficient for taking optimal nutrition. 3) Calorie, carbohydrate, vitamin B, and niacin were taken nutrition from a staple food and protein from at similar ratio of staple and subsidiary food and fat, Fe, vitamin A, vitamin B1, adn vitamin C were mainly taken from a staple food. It was indicated that B-Group was higher at the supply-ratio by a staple food than A-Group. 3. The realities of improvement of physical conditions The improvement of physical conditions of the two groups were lower in accordance with Korea Children's Growth Standard, but B-Group was inferior to A-Group. 4. Seeing that the total intake of nutrition being taken at lunch time was lower as campared to the RDA intakes between the two Groups were remakably different, a lot of efforts should be made to improve the calorie-intake for the purpose of the farm and fishing villages. For balanced intake of nutrition, selection of foods and various cooking ways are important, therefore, throughout the expansion of the lowest stratum of elementary schools for milk-supply, high-quality protein, fat, Ca, vitamin B2 and etc should be supplied to the growing students. That can narrow the gaps of the intake of nutrition and physical condition between the students who live in cities and the students who live in the country.

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Triptolide Inhibits the Proliferation of Immortalized HT22 Hippocampal Cells Via Persistent Activation of Extracellular Signal-Regulated Kinase-1/2 by Down-Regulating Mitogen-Activated Protein Kinase Phosphatase-1 Expression

  • Koo, Hee-Sang;Kang, Sung-Don;Lee, Ju-Hwan;Kim, Nam-Ho;Chung, Hun-Taeg;Pae, Hyun-Ock
    • Journal of Korean Neurosurgical Society
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    • v.46 no.4
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    • pp.389-396
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    • 2009
  • Objective : Triptolide (TP) has been reported to suppress the expression of mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1), of which main function is to inactivate the extracellular signal-regulated kinase-1/2 (ERK-1/2), the p38 MAPK and the c-Jun N-terminal kinase-1/2 (JNK-1/2), and to exert antiproliferative and pro-apoptotic activities. However, the mechanisms underlying antiproliferative and pro-apoptotic activities of TP are not fully understood. The purpose of this study was to examine whether the down-regulation of MKP-1 expression by TP would account for antiproliferative activity of TP in immortalized HT22 hippocampal cells. Methods : MKP-1 expression and MAPK phosphorylation were analyzed by Western blot. Cell proliferation was assessed by $^3H$-thymidine incorporation. Small interfering RNA (siRNA) against MKP-1, vanadate (a phosphatase inhibitor), U0126 (a specific inhibitor for ERK-1/2), SB203580 (a specific inhibitor for p38 MAPK), and SP600125 (a specific inhibitor for JNK-1/2) were employed to evaluate a possible mechanism of antiproliferative action of TP. Results : At its non-cytotoxic dose, TP suppressed MKP-1 expression, reduced cell growth, and induced persistent ERK-1/2 activation. Similar growth inhibition and ERK-1/2 activation were observed when MKP-1 expression was blocked by MKP-1 siRNA and its activity was inhibited by vanadate. The antiproliferative effects of TP, MKP-1 siRNA, and vanadate were significantly abolished by U0126, but not by SB203580 or SP600125. Conclusion : Our findings suggest that TP inhibits the growth of immortalized HT22 hippocampal cells via persistent ERK-1/2 activation by suppressing MKP-1 expression. Additionally, this study provides evidence supporting that MKP-1 may play an important role in regulation of neuronal cell growth.

A Study on the Mechanism of Calcium Binding Inhibition of Cardiac Sarcoplasmic Reticulum by Oxygen Free Radicals (산소대사물에 의한 심장근 Sarcoplasmic reticulum의 칼슘운반 억제 기전에 관한연구)

  • Kim, Hae-Won;Chung, Myung-Hee;Kim, Myung-Suk;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.21 no.2
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    • pp.79-89
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    • 1985
  • Mechanism of calcium transport inhibition of cardiac sarcoplasmic reticulum (SR) by oxygen free radicals was examined. Effects of oxygen free radicals generated by xanthine/xanthine oxidase (X/XO) system on isolated porcine ventricle SR were studied with respect to its calcium binding, lipid peroxidation, SH-group content and alteration of membrane protein components. The results are as follows. 1) Calcium binding of isolated SR was markedly inhibited by X/XO. 2) During the incubation of sarcoplasmic reticulum with xanthine/xanthine oxidase, there were marked inclose in lipid peroxidation and reduction of SH-group content. 3) An antioxidant, p-phenylenediamine effectively prevented the lipid peroxidation but partially prevented the calcium binding inhibition of X/XO treated SR. 4) The reduction of SH-group content of SR treated with X/XO was partially prevented by p-phenylendiamine. 5) When modifying SH-group of SR by treatment with DTNB, the inhibition of calcium binding activity was partially prevented. 6) On gel-permeation chromatography of X/XO-treated sarcoplasmic reticulum, there was an increase of small molecular weight products, probably protein degradation products. 7) Semicarbazide, which prevents the cross-linking reaction of protein components, did not affect the calcium binding inhibition of X/XO-treated SR. From these results, it is suggested that the inhibition of calcium binding of SR by oxygen free radicals results from the consequence of multiple changes of SR components, which are lipid peroxidation, SH-group oxidation and degradation of protein components.

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Intracellular Signaling Pathway for Host Defense Mechanisms against Piscine Nervous Necrosis Virus (NNV) (어류신경괴사증바이러스(nervous necrosis virus, NNV) 감염에 따른 숙주의 방어기전관련 세포신호전달)

  • Kim, Jong-Oh
    • Journal of Life Science
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    • v.30 no.4
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    • pp.402-409
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    • 2020
  • Nervous necrosis virus (NNV) contains a bi-segmented viral genome, RNA1 (3.4 kb, RdRp), and RNA2 (1.4 kb, capsid protein) in a small particle (25 nm). Despite its extremely compact size, NNV has caused serious damage by infecting approximately 120 fish species worldwide since it was first reported in the late 1980s. In order to minimize the damage caused by NNV infection and develop effective vaccines, it is necessary to understand the intra cellular signaling system according to NNV infection. NNV infection induces cell cycle arrest at the G1 phase via the p53-dependent pathway to use the cellular system for its replication. Otherwise, host cells recognize NNV infection through the RIG-1-like receptor (RLR) signaling pathway to control the virus and infected cells, and then ISGs required for antiviral action are activated via the IFN signaling pathway. Moreover, apoptosis of infected cells is triggered by the unfolded protein response (UPR) through ER stress and mitochondria-mediated cell death. Cell signaling studies on the NNV infection mechanisms are still at an early stage and many pathways have yet to be identified. Understanding the various disease-specific cellular signaling systems associated with NNV infection is essential for rapid and accurate diagnosis and vaccine development.

Classification of Antimicrobial Peptides among the Innate Immune Modulators (선천성 면역조절자인 항생펩타이드 분류)

  • Lee, Jong-Hwan
    • Journal of Life Science
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    • v.25 no.7
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    • pp.833-838
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    • 2015
  • Multidrug-resistant super bacterial, fungal, viral, and parasitic infections are major health threaten pathogens. However, to overcome the present healthcare situation, among the leading alternatives to current drugs are antimicrobial peptides (AMPs), which are abundantly produced via various species in nature. AMPs, small host defense proteins, are in charge of the innate immunity for the protection of multicellular organisms such as fish, amphibian, reptile, plants and animals from infection. The number of AMPs identified per year has increased steadily since the 1980s. Over 2,000 natural AMPs from bacteria, protozoa, fungi, plants, and animals have been listed into the antimicrobial peptide database (APD). The majority of these AMPs (>86%) possess 11–50 amino acids with a net charge from 0 to +7 and hydrophobic percentages between 31–70%. This report classified AMP into several categories including biological source, biological functions, peptide properties, covalent bonding pattern, and 3D structure. AMP functions not only antimicrobial activity but facilitates cell biological activity such as chemotatic activity. In addition, fibroblastic reticular cell (FRC) originated from mouse lymph node stroma induced the expression of AMP in inflammatory condition. AMP induced from FRC contained whey acidic protein (WAP) domain. It suggests that the classification of AMP will be done by protein domain.

Inhibition of L-type Ca2+ current by ginsenoside Rd in rat ventricular myocytes

  • Lu, Cheng;Sun, Zhijun;Wang, Line
    • Journal of Ginseng Research
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    • v.39 no.2
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    • pp.169-177
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    • 2015
  • Background: Ginsenoside Rd (GSRd), one of the most abundant ingredients of Panax ginseng, protects the heart via multiple mechanisms including the inhibition of $Ca^{2+}$ influx.We intended to explore the effects of GSRd on L-type $Ca^{2+}$ current ($I_{Ca,L}$) and define the mechanism of the suppression of $I_{Ca,L}$ by GSRd. Methods: Perforated-patch recording and whole-cell voltage clamp techniques were applied in isolated rat ventricular myocytes. Results: (1) GSRd reduced $I_{Ca,L}$ peak amplitude in a concentration-dependent manner [half-maximal inhibitory concentration $(IC_{50})=32.4{\pm}7.1{\mu}mol/L$] and up-shifted the current-voltage (I-V) curve. (2) GSRd ($30{\mu}mol/L$) significantly changed the steady-state activation curve of $I_{Ca,L}$ ($V_{0.5}:-19.12{\pm}0.68$ vs. $-6.26{\pm}0.38mV$; n = 5, p < 0.05) and slowed down the recovery of $I_{Ca,L}$ from inactivation [the time content (${\zeta}$) from 91 ms to 136 ms, n = 5, p < 0.01]. (3) A more significant inhibitive effect of GSRd ($100{\mu}mol/L$) was identified in perforated-patch recording when compared with whole-cell recording [$65.7{\pm}3.2%$ (n = 10) vs. $31.4{\pm}5.2%$ (n = 5), p < 0.01]. (4) Pertussis toxin ($G_i$ protein inhibitor) completely abolished the $I_{Ca,L}$ inhibition induced by GSRd. There was a significant difference in inhibition potency between the two cyclic adenosine monophosphate elevating agents (isoprenaline and forskolin) prestimulation [$55{\pm}7.8%$ (n = 5) vs. $17.2{\pm}3.5%$ (n = 5), p < 0.01]. (5) 1H-[1,2,4]Oxadiazolo[4,3-a]-quinoxalin-1-one (a guanylate cyclase inhibitor) and N-acetyl-$\small{L}$-cysteine (a nitric oxide scavenger) partly recovered the $I_{Ca,L}$ inhibition induced by GSRd. (6) Phorbol-12-myristate-13-acetate (a protein kinase C activator) and GF109203X (a protein kinase C inhibitor) did not contribute to the inhibition of GSRd. Conclusion: These findings suggest that GSRd could inhibit $I_{Ca,L}$ through pertussis toxin-sensitive G protein ($G_i$) and a nitric oxide-cyclic guanosine monophosphate-dependent mechanism.

Characteristics of Tenebrio molitor larvae and Bombyx mori pupae sequentially fermented with Aspergillus oryzae and Bacillus subtilis (Aspergillus oryzae와 Bacillus subtilis를 순차접종 발효한 갈색거저리유충과 누에번데기 발효물의 품질특성)

  • Kang, Sanghun;Chung, Chang-Ho
    • Korean Journal of Food Science and Technology
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    • v.54 no.1
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    • pp.94-102
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    • 2022
  • Defatted soybean, larvae of brown mealworm (Tenebrio molitor), and powdered pupae of silkworm (Bombyx mori) were fermented in solid and liquid forms using Aspergillus oryzae and Bacillus subtilis. The protein degradation rate (NDR) through solid fermentation was the highest in the fermented soybean control sample (54.69±6.54%), followed by silkworm pupae (34.82±5.99%) and brown mealworm larvae (30.54±3.80%). When these edible insects were fermented in liquid form, solid extraction yield was 37.73-46.88%, and protein yield was 47.47-63.02%. NDR of fermented liquid form products increased to 58.90, 52.62, and 50.13% for soybean, brown mealworm larvae, and silkworm pupae, respectively. SDS-PAGE of the liquid fermented products confirmed that microbial fermentation decomposed higher-molecular-weight proteins into small polypeptides. In vitro digestibility of liquid forms of edible insects increased by 1.26 to 1.53 times after fermentation. The protein solubility, foaming ability, and foam stability of liquid-fermented edible insects all tended to increase through fermentation.

Comparison of miR-106b, miR-191, and miR-30d expression dynamics in milk with regard to its composition in Holstein and Ayrshire cows

  • Marina V. Pozovnikova;Viktoria B. Leibova;Olga V. Tulinova;Elena A. Romanova;Artem P. Dysin;Natalia V. Dementieva;Anastasiia I. Azovtseva;Sergey E. Sedykh
    • Animal Bioscience
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    • v.37 no.6
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    • pp.965-981
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    • 2024
  • Objective: Milk composition varies considerably and depends on paratypical, genetic, and epigenetic factors. MiRNAs belong to the class of small non-coding RNAs; they are one of the key tools of epigenetic control because of their ability to regulate gene expression at the post-transcriptional level. We compared the relative expression levels of miR-106b, miR-191, and miR-30d in milk to demonstrate the relationship between the content of these miRNAs with protein and fat components of milk in Holstein and Ayrshire cattle. Methods: Milk fat, protein, and casein contents were determined in the obtained samples, as well as the content of the main fatty acids (g/100 g milk), including: saturated acids, such as myristic (C14:0), palmitic (C16:0), and stearic (C18:0) acids; monounsaturated acids, including oleic (C18:1) acid; as well as long-, medium- and short-chain, polyunsaturated, and trans fatty acids. Real-time stem-loop one-tube reverse transcription polymerase chain reaction with TaqMan probes was used to measure the miRNA expression levels. Results: The miRNA expression levels in milk samples were found to be decreased in the first two months in Holstein breed, and in the first four months in Ayrshire breed. Correlation analysis did not reveal any dependence between changes in the expression level of miRNA and milk fat content, but showed a multidirectional relationship with individual milk fatty acids. Positive associations between the expression levels of miR-106b and miR-30d and protein and casein content were found in the Ayrshire breed. Receiver operating characteristic curve analysis showed that miR-106b and miR-30d expression levels can cause changes in fatty acid and protein composition of milk in Ayrshire cows, whereas miR-106b expression level determines the fatty acid composition in Holsteins. Conclusion: The data obtained in this study showed that miR-106b, miR-191, and miR-30d expression levels in milk samples have peculiarities associated with breed affiliation and the lactation period.

FUN14 Domain-Containing Protein 1 Is Involved in Amyloid Beta Peptide-Induced Mitochondrial Dysfunction and Cell Injury in HT-22 Neuronal Cells (HT-22 신경세포에서 아밀로이드 베타 펩티드에 의한 미토콘드리아와 세포 손상 기전에서 FUN14 도메인 함유 단백 1의 역할)

  • Jae Hoon Kang;Jae Suk Woo
    • Journal of Life Science
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    • v.34 no.1
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    • pp.37-47
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    • 2024
  • FUN14 domain-containing protein 1 (FUNDC1), an outer mitochondrial membrane protein, contributes to removal of damaged mitochondria through mitophagy. In this study, to elucidate the role of the FUNDC1 in the amyloid beta peptide (Aβ)-induced neuropathy, changes in the degree of mitochondrial dysfunction and cell injury caused by Aβ treatment were examined in the HT-22 neuronal cells in which the FUNDC1 expression was transiently silenced or overexpressed. We found that Aβ treatment causes a time-dependent decrease of the FUNDC1 expression. In the Aβ-treated cells, there were a drop in MTT reduction ability, depletion of cellular ATP, disruption of mitochondrial membrane potential, stimulation of cellular ROS production, and increased mitochondrial Ca2+ load. Activation of caspase-3 and induction of apoptotic cell death were also observed. Transient silencing of the FUNDC1 expression by transfection with the FUNDC1 small interfering RNA per se caused mitochondrial dysfunction and apoptotic cell death like the effect of Aβ treatment. Conversely, in cells in which the FUNDC1 was transiently overexpressed by FUNDC1-Myc transfection, overexpression itself had no effect on the mitochondrial functional integrity and cell survival but showed a significant prevention effect against mitochondrial and cell injury caused by Aβ treatment. Overall, these results suggest that the FUNDC1 is importantly involved in the Aβ-induced mitochondrial dysfunction and cell injury in the HT-22 neuronal cells.