• Title/Summary/Keyword: industrial proteomics

검색결과 29건 처리시간 0.027초

Siamese Crocodile White Blood Cell Extract Inhibits Cell Proliferation and Promotes Autophagy in Multiple Cancer Cell Lines

  • Phosri, Santi;Jangpromma, Nisachon;Chang, Leng Chee;Tan, Ghee T.;Wongwiwatthananukit, Supakit;Maijaroen, Surachai;Anwised, Preeyanan;Payoungkiattikun, Wisarut;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • 제28권6호
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    • pp.1007-1021
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    • 2018
  • Cancer represents one of the most significant threats to human health on a global scale. Hence, the development of effective cancer prevention strategies, as well as the discovery of novel therapeutic agents against cancer, is urgently required. In light of this challenge, this research aimed to evaluate the effects of several potent bioactive peptides and proteins contained in crocodile white blood cell extract (cWBC) against LU-1, LNCaP, PC-3, MCF-7, and CaCo-2 cancer cell lines. The results demonstrate that 25, 50, 100, and $200{\mu}g/ml$ cWBC exhibits a strong cytotoxic effect against all investigated cell lines ($IC_{50}$ $70.34-101.0{\mu}g/ml$), while showing no signs of cytotoxicity towards noncancerous Vero and HaCaT cells. Specifically, cWBC treatment caused a significant reduction in the cancerous cells' colony forming ability. A remarkable suppression of cancerous cell migration was observed after treatment with cWBC, indicating potent antimetastatic properties. The mechanism involved in the cancer cell cytotoxicity of cWBC may be related to apoptosis induction, as evidenced by typical apoptotic morphology features. Moreover, certain cWBC concentrations induced significant overproduction of ROS and significantly inhibited the $S-G_2/M$ transition in the cancer cell. The molecular mechanisms of cWBC in apoptosis induction were to decrease Bcl-2 and XIAP expression levels and increase the expression levels of caspase-3, caspase-8, and p53. These led to a decrease in the expression level of the cell cycle-associated gene cyclin-B1 and the arrest of cell population growth. Consequently, these findings demonstrate the prospect of the use of cWBC for cancer therapy.

New Protein Extraction/Solubilization Protocol for Gel-based Proteomics of Rat (Female) Whole Brain and Brain Regions

  • Hirano, Misato;Rakwal, Randeep;Shibato, Junko;Agrawal, Ganesh Kumar;Jwa, Nam-Soo;Iwahashi, Hitoshi;Masuo, Yoshinori
    • Molecules and Cells
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    • 제22권1호
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    • pp.119-125
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    • 2006
  • The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using precast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins. Our results reveal enrichment of protein spots by use of individual brain regions rather than whole brain, as well as the presence of differentially expressed spots in their proteomes. Thus individual brain regions provide improved protein coverage and are better suited for differential protein detection. Moreover, using a phosphoprotein-specific dye, ingel detection of phosphoproteins was demonstrated. Representative high-resolution silver nitrate stained proteome profiles of rat whole brain total soluble protein are presented. Shortcomings apart (failure to separate membrane proteins), gel-based proteomics remains a viable option, and 2-DGE is the method of choice for generating high-resolution proteome maps of rat brain and brain regions.

In Vivo Wound Healing Activity of Crocodile (Crocodylus siamensis) Hemoglobin and Evaluation of Antibacterial and Antioxidant Properties of Hemoglobin and Hemoglobin Hydrolysate

  • Pakdeesuwan, Anawat;Araki, Tomohiro;Daduang, Sakda;Payoungkiattikun, Wisarut;Jangpromma, Nisachon;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.26-35
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    • 2017
  • The hydrolysis of proteins constitutes an invaluable tool, granting access to a variety of peptide fragments with potentially interesting biological properties. Therefore, a hemoglobin (Hb) hydrolysate of Crocodylus siamensis was generated by digestion under acidic conditions. The antibacterial and antioxidant activities of the Hb hydrolysate were assessed in comparison with intact Hb. A disc diffusion assay revealed that the Hb hydrolysate exhibited antibacterial activity against eight strains of gram-positive bacteria and showed a higher efficacy than intact Hb. Moreover, the antioxidant activity of intact Hb and its hydrolysate was evaluated using ABTS and DPPH radical scavenging assays. The Hb hydrolysate exhibited free radical scavenging rates of 6-32%, whereas intact Hb showed a slightly higher activity. In addition, non-toxicity to human erythrocytes was observed after treatment with quantities of Hb hydrolysate up to $10{\mu}g$. Moreover, active fragmented Hb (P3) was obtained after purifying the Hb hydrolysate by reversed-phase HPLC. Scanning electron microscopy demonstrated the induction of bacterial cell membrane abnormalities after exposure to P3. Antibacterial and antioxidant activities play crucial roles for supporting the wound healing activity. Consequently, an in vivo mice excisional skin wound healing assay was carried out to investigate the effects of intact Hb treatment on wound healing in more detail. The results clearly demonstrate that intact Hb is capable of promoting 75% wound closure within 6 days. These findings imply that intact Hb of C. siamensis and its acid hydrolysate may serve as valuable precursors for food supplementary products benefitting human health.

In Vitro and in Vivo Wound Healing Properties of Plasma and Serum from Crocodylus siamensis Blood

  • Jangpromma, Nisachon;Preecharram, Sutthidech;Srilert, Thanawan;Maijaroen, Surachai;Mahakunakorn, Pramote;Nualkaew, Natsajee;Daduang, Sakda;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • 제26권6호
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    • pp.1140-1147
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    • 2016
  • The plasma and serum of Crocodylus siamensis have previously been reported to exhibit potent antimicrobial, antioxidant, and anti-inflammatory activities. During wound healing, these biological properties play a crucial role for supporting the formation of new tissue around the injured skin in the recovery process. Thus, this study aimed to evaluate the wound healing properties of C. siamensis plasma and serum. The collected data demonstrate that crocodile plasma and serum were able to activate in vitro proliferation and migration of HaCaT, a human keratinocyte cell line, which represents an essential phase in the wound healing process. With respect to investigating cell migration, a scratch wound experiment was performed which revealed the ability of plasma and serum to decrease the gap of wounds in a dose-dependent manner. Consistent with the in vitro results, remarkably enhanced wound repair was also observed in a mouse excisional skin wound model after treatment with plasma or serum. The effects of C. siamensis plasma and serum on wound healing were further elucidated by treating wound infections by Staphylococcus aureus ATCC 25923 on mice skin coupled with a histological method. The results indicate that crocodile plasma and serum promote the prevention of wound infection and boost the re-epithelialization necessary for the formation of new skin. Therefore, this work represents the first study to demonstrate the efficiency of C. siamensis plasma and serum with respect to their wound healing properties and strongly supports the utilization of C. siamensis plasma and serum as therapeutic products for injured skin treatment.

전산학의 최신 응용 및 학제 분야인 생명정보학 (Bioinformatics : Latest Application and Interdisciplinary Field of Computer Science)

  • 김기봉
    • 한국산학기술학회논문지
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    • 제11권3호
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    • pp.971-977
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    • 2010
  • 본 바이오데이터들이 홍수처럼 쏟아지면서 컴퓨터를 활용해 해결해야 할 많은 문제들이 야기되고 있다. 생체분자들과 연관된 정보들을 대량으로 분석하는 전산기술 응용분야인 생명정보학은 학제간 학문분야로서 현재 확고히 자리매김하고 있으며. 구조생물학, 유전체학, 단백질체학, 시스템 생물학, 생물통계학 및 전산학 등 광범위한 학문영역을 포괄한다. 본 총설에서는 생명정보학에 대한 최근 상황과 전반적인 응용분야에 대해 소개하고자 한다. 일반적으로 널리 사용되는 생명정보 및 바이오데이터베이스들의 유형을 살펴보고, 전산학 분야와 긴밀한 관계가 있는 몇가지 생명정보학 응용 분야들을 다루고자 한다.

Expression of P-glycoprotein is Positively Correlated with p53 in Human Papilloma Virus Induced Squamous Intraepithelial Lesions of Uterine Cervix: Poor Prognosis Association

  • Singh, Madhulika;Singh, Uma;Mathur, Neeraj;Shukla, Yogeshwer
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6039-6045
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    • 2012
  • This study was conducted to assess the predictive value of p-glycoprotein (p-gp) and p53 immunoexpression in human papillomavirus (HPV) infected cases of cervical dysplasia. Expression of both p-gp and p53 proteins was detected in cervical smears from 177 squamous intraepithelial lesions (SIL) cases along with 183 "atypical squamous cells of unknown significance" (ASCUS) and 150 normal cases. HPV 16 and 18 infection was detected by polymerase chain reaction using type-specific primers for HPV sub-types. There were no significant detectable p53 and p-gp expression in the normal cervix smears (p>0.05). In the ASCUS group 10 cases were positive for both p53 and p-gp immunoreactivity. In cervical dysplasia cases, p53 was positive in 86 (48.58%) while p-gp was positive in 93 (52.54%) and the two markers showed a highly significant correlation (r=0.92, p<0.001). Expression of p53 and p-gp was associated with grade of SIL (p<0.001). A positive correlation between the presence of HPV and expression of proteins p53 and p-gp in smears of patients with cervical lesions was also noted (p<0.001). Thus, p53 and p-gp immunostaining in cervical smears may act as an auxiliary biomarker for detection of HPV-associated cervical lesions. Additionally, a significant positive correlation between ascending grades of SIL and labeling indices of markers suggests that p53 and p-gp can be used as an adjunct to cytomorphological interpretation of conventional cervical Pap smears.

Molecular Cloning and Expression of a Novel Cuticle Protein Gene from the Chinese Oak Silkmoth, Antheraea pernyi

  • Kim Bo Yeon;Park Nam Sook;Jin Byung Rae;Kang Pil Don;Lee Bong Hee;Seong Su Il;Hwang Jae Sam;Chang Jong Su;Lee Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
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    • 제10권1호
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    • pp.11-17
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    • 2005
  • In our research to identify gene involved in the cuticle protein, we cloned a novel cuticle protein gene, ApCP15.5, from the Chinese oak silkmoth, Antheraea pernyi, larvae cDNA library. The gene encodes a 149 amino acid polypeptide with a predicted molecular mass of 15.5 kDa and a pI of 9.54. The ApCP15.5 contained a type-specific consensus sequence identifiable in other insect cuticle proteins and the deduced amino acid sequence of the ApCP15.5 cDNA is most homologous to Tenebrio molitor-C1B ($43\%$ protein sequence identity), followed by Locusta migratoria-76 ($42\%$ protein sequence identity). Northern blot and Western blot analyses revealed that the ApCP15.5 showed the epidermis-specific expression. The expression profile of ApCP15.5 indicated that the ApCP15.5 mRNA expression was detected in the early stages after larval ecdysis and larval-pupal metamorphosis, and its expression level was most significant on the first day of larval ecdysis and pupal stage. The ApCP15.5 was expressed as a 15.5 kDa polypeptide in baculovirus-infected insect cells.

일부 자원자를 통한 납, 카드뮴의 다경로 인체 노출평가 (Human Multi-route Exposure Assessment of Lead and Cadmium for Korean Volunteers)

  • 오은하;임호섭;장재연;이은일
    • Journal of Preventive Medicine and Public Health
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    • 제39권1호
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    • pp.53-58
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    • 2006
  • Objectives: We wanted to evaluate the main route of exposure to lead and cadmium for the general population in Korea by using multi-route and multi-media exposure assessment Methods: Samples of air at the homes, samples of the food and water and peripheral blood samples were collected from thirty volunteers living in Seoul (the metropolitan area), Yong-in (the suburban area) and Ansan (the industrial area) in 2001. Graphite furnace atomic absorption spectrometric methods were used for the determination of the Pb and Cd levels in the air, food, water and blood samples. Results: The average intake of lead through the air, drinking water and food were $5.06{\mu}g/day\;(26.3%)\;0.002{\mu}g/day\;(0.1%),\;and\;16.4{\mu}g/day\;(73.6%)$, respectively. The average intake of cadmium through the air, drinking water and food were $0.082{\mu}g/day\;(0.9%),\;0.001{\mu}g/day\;(0.007%),\;and\;12.61 {\mu}g/day\;(99.0%)$, respectively. The blood lead level was statistically higher in the male subjects than in the female subjects (3.39 and $2.22{\mu}g/dl$, respectively), and only gender was a significant variable on the multiple regression analysis for blood lead. Conclusions: This study showed that the intake of lead and cadmium through food was the major route of exposure. A policy to reduce the pollutants according to the exposure routes should be established. However, more studies will be needed to support these data for the general population.