• 제목/요약/키워드: Biological screening methods

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자궁경부암세포에서 방사선조사시 차등 발현되는 유전자 동정 (Identification of Differentially Expressed Radiation-induced Genes in Cervix Carcinoma Cells Using Suppression Subtractive Hybridization)

  • 김준상;이영숙;이증훈;이웅희;성은영;조문준
    • Radiation Oncology Journal
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    • 제23권1호
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    • pp.43-50
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    • 2005
  • 목적 : 자경경부암세포에서 polymeric chain reaction (PCR)원리를 이용한 suppression subtractive hybridization (SSH) 방법으로 방사선조사 시 차등 발현되는 유전자를 동정하고자 하였다. 대상 및 방법 : 자궁경부암세포주인 HeLa세포주에 방사선조사 전과 후 총 RNA와 poly $(A)^+$ mRNA를 분리였다. SSH방법으로 forward 및 reverse-subtracted cDNA libraries를 만들었다. 차등 발현된 유전자를 screening하기 위해 reverse Northern blotting (dot blot analysis)을 이용하여 각각의 library에서 88개의 클론을 선택하였고 Nothern blotting으로 확인 후 sequencing하였다. 결과 : screening상 176개 클론 중 forward-subtracted library에서 10개의 유전자가 reverse-subtracted library에서 9개의 유전자가 동정되었다. forward-subtracted library로부터 3개의 유전자가 Northern blotting에 의하여 확인되었고 이중 telomerase catalytic subunit and sodium channel-like protein 유전자와 1개의 ESTs (expressed sequence tags) 유전자가 방사선선량에 따라 증가하쳐다. 결론 : 본 연구를 통해 자궁경부암세포주에서 방사선에 의해 유도되는 유전자를 SSH 방법을 통해 동정할 수 있었다. 그러나 이러한 유전자가 어떤 생물학적인 기능을 갖고 있는지에 대한 계속적인 연구가 필요하다

Screening of Differentially Expressed Genes Related to Bladder Cancer and Functional Analysis with DNA Microarray

  • Huang, Yi-Dong;Shan, Wei;Zeng, Li;Wu, Yang
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권8호
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    • pp.4553-4557
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    • 2013
  • Objective: The purpose of this study was to identify genes related to bladder cancer with samples from normal and disease cases by microarray chip. Methods: After downloading the gene expression profile GSE3167 from Gene Expression Omnibus database which includes 50 bladder samples, comprising 9 normal and 41 disease samples, differentially expressed genes were identified with packages in R language. The selected differentially expressed genes were further analyzed using bioinformatics methods. Firstly, molecular functions, biological processes and cell component analysis were researched by software Gestalt. Then, software String was used to search interaction relationships among differentially expressed genes, and hub genes of the network were selected. Finally, by using plugins of software Cytoscape, Mcode and Bingo, module analysis of hub-genes was performed. Results: A total of 221 genes were identified as differentially expressed by comparing normal and disease bladder samples, and a network as well as the hub gene C1QBP was obtained from the network. The C1QBP module had the closest relationship to production of molecular mediators involved in inflammatory responses. Conclusion: We obtained differentially expressed genes of bladder cancer by microarray, and both PRDX2 and YWHAZ in the module with hub gene C1QBP were most significantly related to production of molecular mediators involved in inflammatory responses. From knowledge of inflammatory responses and cancer, our results showed that, the hub gene and its module could induce inflammation in bladder cancer. These related genes are candidate bio-markers for bladder cancer diagnosis and might be helpful in designing novel therapies.

단백질(蛋白質) 품질평가(品質評價)를 위(爲)한 신속방법(迅速方法) (Rapid In Vitro Methods for Protein Evaluation)

  • 류홍수;이강호
    • 한국식품영양과학회지
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    • 제14권2호
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    • pp.202-213
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    • 1985
  • The protein nutritional quality of foods has become an important factor to food processors with the advent of nutritional labeling regulations for foods. Then, as is true today, the officially approved assay for protein nutritional quality was the rat based protein efficiency ratio(PER) bioassay. The PER bioassay requires a minimum of 28 days to performe, and is therefore not applicable to routine quality assurance use by the food industry. Within the past ten years there has been a research emphasis placed on the development of rapid, inexpensive, biological and/or chemical based assays for protein nutritional quality. It was hoped that if a rapid assay could be developed and thoroughly tested, it could be used in lieu of the PER bioassay in the day-to-day quality assurance screening of food ingredients and products. The rapid assays developed in the hope of attaining this goal have been based on microorganisms, proteolytic enzymes, and amino acid profiles, as well as combinations of the above. In this review, it will be described and briefly discussed many of procedures which had contributed conceptually as well as practically to the development of in vitro methods for the evaluation of protein quality. Special emphasis will be placed on the C-PER(computed protein efficiency ratio) assay which combines data from in vitro protease digestion and amino acid composition to predict protein nutritional quality designed by Satterlee et al. (1980), and the DC-PER(discriminant computed PER) which is a method of estimating protein quality based on rat assay and in vitro digestibility obtained using solely essential amino acid data will be also introduced.

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Assessment of Masks Used by Healthcare Workers: Development and Validation of a Mask Qualitative Assessment Tool (MQAT)

  • Gharibi, Vahid;Cousins, Rosanna;Mokarami, Hamidreza;Jahangiri, Mehdi;Keshavarz, Mohammad A.;Shirmohammadi-Bahadoran, Mohammad M.
    • Safety and Health at Work
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    • 제13권3호
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    • pp.364-371
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    • 2022
  • Background: Respiratory masks can provide healthcare workers with protection from biological hazards when they have good performance. There is a direct relationship between the visual specifications of a mask and its efficacy; thus, the aim of this study was to develop tools for qualitative assessment of the performance of masks used by healthcare workers. Methods: A mixed-methods design was used to develop a qualitative assessment tool for medical face masks (MFM) and particle filtering half masks (PFHM). The development of domains and items was undertaken using observation and interviews, the opinions of an expert panel, and a review of texts and international standards. The second phase evaluated the psychometric properties of tools. Finally, the validated Mask Qualitative Assessment Tools (MQAT) were used to assess six samples from 10 brands of the two types of masks. Results: MQAT-MFM and MQAT-PHFM shared 42 items across seven domains: "cleanliness," "design," "marking, labeling and packaging," "mask layers," "mask strap," "materials and construction," and "nose clip." MQAT-MFM included one additional item. MQAT-PHFM included another nine items associated with an eighth "Practical Performance" domain, and the valve version had another additional "Exhalation Valve" domain and six items. The evaluation indicated 80% compliance for MFM and 71% compliance for PFHM. "Marking, labeling and packaging" and "Layers" were associated with the least compliance in both types of masks and should be checked carefully for defining mask quality. Conclusion: MQAT can be used for immediate screening and initial assessment of MFM and PHFM through appearance, simple tools, and visual inspection.

크롬 직업병 유소견자와 요관찰자 추이 및 취급 근로자의 생물학적 노출수준 고찰 (Review of the Literature on Level of Biological Exposure and Trends in Possible and Probable Occupational Disease among Chromium Workers)

  • 박상일;김남수;황보영;김화성;이성수;김용배
    • 한국산업보건학회지
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    • 제31권3호
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    • pp.213-225
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    • 2021
  • Objectives: This study identifies the trends in possible and probable occupational disease among chromium workers and reviews the literature on domestic and foreign chromium workers to review the industries, biological exposure levels, and major results. Methods: The results of the Ministry of Employment and Labor's special health-screening program for hazard agents among workers from 2009 to 2019 were used. Also, the industries, biological exposure levels, and major results on chromium workers were reviewed using PubMed and RISS. Results: The average annual proportion of both possible and probable occupational disease for chromium workers has recently increased. The average annual proportion of possible and probable occupational disease that can occur was high for both men and women in their 60s or older by age and 10 to 14 years by work duration. By industry, possible occupational disease showed high in manufacturing. In the literature review, many electroplating-related chromium-workers reported high levels of exposure to blood and urine chromium, as did dental technicians; tannery, tile factory or glass mill workers; cement workers; and sodium bichromate workers. Furthermore, a number of main findings in recent studies have reported that chromium exposure is related to genetic toxicity among workers. Conclusions: In this study, the average annual rate of both possible and probable occupational disease in domestic chromium workers is increasing, and a body of literature shows that chromium exposure is related to genetic toxicity and associated indicators among workers, which requires more systematic study.

Phytochemical, Antidiabetic, Antioxidant, Antibacterial, Acute and Sub-Chronic Toxicity of Moroccan Arbutus unedo Leaves

  • Latifa Doudach;Hanae Naceiri Mrabti;Samiah Hamad Al-Mijalli;Mohamed Reda Kachmar;Kaoutar Benrahou;Hamza Assaggaf;Ahmed Qasem;Emad Mohamed Abdallah;Bodour Saeed Rajab;Khouloud Harraqui;Mouna Mekkaoui;Abdelhakim Bouyahya;Moulay El Abbes Faouzi
    • 대한약침학회지
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    • 제26권1호
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    • pp.27-37
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    • 2023
  • Objectives: Moroccan Arbutus unedo is an essential medicinal plant; however, little is known about the biological properties of its leaves mentioned in Moroccan traditional medicine. Methods: Various standard experiments were performed to evaluate the phytochemical, antidiabetic, antioxidant, antibacterial, and acute and sub-chronic toxicity characteristics of A. unedo leaves. Results: Phytochemical screening led to the identification of several phytochemical classes, including tannins, flavonoids, terpenoids, and anthraquinones, with high concentrations of polyphenols (31.83 ± 0.29 mg GAEs/g extract) and flavonoids (16.66 ± 1.47 mg REs/g extract). Further, the mineral analysis revealed high levels of calcium and potassium. A. unedo extract demonstrated significant antioxidant and anti-diabetic activities by inhibiting α-amylase (1.350 ± 0.32 g/mL) and α-glucosidase (0.099 ± 1.21 g/mL) compared to the reference drug Acarbose. Also, the methanolic extract of the plant exhibited significantly higher antibacterial activity than the aqueous extract. Precisely, three of the four examined bacterial strains exhibited substantial susceptibility to the methanolic extract . Minimum bactericidal concentration (MBC)/minimum inhibitory concentration (MIC) values indicated that A. unedo harbor abundant bactericidal compounds. For toxicological studies, mice were administered with A. unedo aqueous extract at single doses of 2,000 and 5,000 mg/kg. They did not exhibit significant abnormal behavior, toxic symptoms, or death during the 14-day acute toxicity test and the 90-day sub-chronic toxicity test periods. The general behavior, body weight, and hematological and biochemical status of the rats were assessed, revealing no toxicological symptoms or clinically significant changes in biological markers observed in the mice models, except hypoglycemia, after 90 days of daily dose administration. Conclusion: The study highlighted several biological advantages of A. unedo leaves without toxic effects in short-term application. Our findings suggest that conducting more comprehensive and extensive in vivo investigations is of utmost importance to identify molecules that can be formulated into pharmaceuticals in the future.

Efficient Macrocyclization for Cyclicpeptide Using Solid-Phase Reaction

  • Kim, Joong-Hup;Hong, Il-Khee;Kim, Hyo-Jeong;Jeong, Hyeh-Jean;Choi, Moon-Jeong;Yoon, Chang-No;Jeong, Jin-Hyun
    • Archives of Pharmacal Research
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    • 제25권6호
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    • pp.801-806
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    • 2002
  • Cyclicpeptides are important targets in peptide synthesis because of their interesting biological properties. Constraining highly flexible linear peptides by cyclization is one of the mostly widely used approaches to define the bioactive conformation of peptides. Cyclic peptides often have increased receptor affinity and metabolic stability over their linear counterparts. We carried out virtual screening experiment via docking in order to understand the interaction between HLE-Human Leukocyte Elastase and ligand peptide and to identify the sequence that can be a target in various ligand peptides. We made cyclic peptides as a target base on Metlle-Phe sequence having affinity for ligand and receptor active site docking. There are three ways to cyclize certain sequences of amino acids such as Met-lie-Phe-Gly-Ile. First is head-to-tail cyclization method, linking between N-terminal and C-terminal. Second method utilizes amino acid side chain such as thiol functional group in Cys, making a thioether bond. The last one includes an application of resin-substituted amino acids in solid phase reaction. Among the three methods, solid phase reaction showed the greatest yield. Macrocyclization of Fmoc-Met-Ile-Phe-Gly-Ile-OBn after cleavage of Fmoc protection in solution phase was carried out to give macrocyclic compound 5 in about 7% yield. In the contrast with solution phase reaction, solid phase reaction for macrocyclization of Met-Ile-Phe-Gly-Ile-Asp-Tentagel in normal concentrated condition gave macrocyclic compound 7 in more than 35% yield.

Salivary soluble receptor activator of nuclear factor kappa B ligand/osteoprotegerin ratio in periodontal disease and health

  • Tabari, Zahra Alizadeh;Azadmehr, Abbas;Tabrizi, Mohammad Amir Alizadeh;Hamissi, Jalaloddin;Ghaedi, Fatemeh Baharak
    • Journal of Periodontal and Implant Science
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    • 제43권5호
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    • pp.227-232
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    • 2013
  • Purpose: The receptor activator of nuclear factor kappa B (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system plays a significant role in osteoclastogenesis, activation of osteoclasts, and regulation of bone resorption. This study aimed to evaluate the use of the salivary soluble RANKL (sRANKL)/OPG ratio as a diagnostic marker for periodontitis in nonsmokers. Methods: Twenty-five patients with chronic periodontitis and 25 individuals with a healthy periodontium were enrolled in this study. Samples containing 5 mL of unstimulated saliva were obtained from each subject. Salivary sRANKL and OPG concentrations were determined using a standard enzyme-linked immunosorbent assay. Statistical analysis was performed using SPSS ver. 18.0. Results: The levels of sRANKL and OPG were detectable in all of the samples. Positive relationships were found between the plaque index and clinical attachment level and both the salivary concentration of sRANKL and the salivary sRANKL/OPG ratio (P<0.05). The salivary concentration of sRANKL and the sRANKL/OPG ratio were significantly higher in the periodontitis group than in the healthy group (P=0.004 and P=0.001, respectively). In contrast, the OPG concentration showed no significant differences between the groups (P=0.455). Conclusions: These findings suggest that the salivary sRANKL/OPG ratio may be helpful in the screening and diagnosis of periodontitis. However, longitudinal studies with larger populations are needed to confirm these results.

In vitro screening of elastase, collagenase, hyaluronidase, and tyrosinase inhibitory and antioxidant activities of 22 halophyte plant extracts for novel cosmeceuticals

  • Jiratchayamaethasakul, Chanipa;Ding, Yuling;Hwang, Ouibo;Im, Seung-Tae;Jang, Yebin;Myung, Seung-Won;Lee, Jeong Min;Kim, Hyun-Soo;Ko, Seok-Chun;Lee, Seung-Hong
    • Fisheries and Aquatic Sciences
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    • 제23권3호
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    • pp.6.1-6.9
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    • 2020
  • Background: Halophyte plant (HPs), a salt-resistant flora, has been reported to provide several health benefits, but the knowledge of its cosmeceutical potential is still ambiguous. Here, 70% ethanol extracts of 22 HPs collected from along the coast of South Korea were investigated for their potentials of antioxidant, anti-aging, and whitening properties for use as materials in novel cosmeceuticals. Methods: Antioxidant activities were determined by DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical and hydrogen peroxide scavenging assays, and skin aging-related enzyme activities (anti-elastase, anti-collagenase, anti-hyaluronidase, and anti-tyrosinase) were evaluated by using the spectrophotometric method. Results: Among the 22 HPs, we found that Ischaemum antephoroides f. coreana and Atriplex gmelinii extracts presented the strongest scavenging effects against DPPH free radical and hydrogen peroxide, respectively. Our finding additionally suggested that Salicornia europaea extract might provide a major source of anti-elastase and anti-hyaluronidase; meanwhile, Rosa rugosa extract showed the highest anti-collagenase effect. Furthermore, the highest tyrosinase inhibitory activity was possessed by Spartina anglica extract. Conclusion: These findings may suggest that halophyte plants showing biological activities may be potent inhibitors of tyrosinase, elastase, collagenase, and hyaluronidase and could be useful for application in cosmeceuticals.

황련지황탕(黃連地黃湯)이 Streptozotocin으로 유발(誘發)된 당뇨병(糖尿病)에 미치는 영향 (Effects of Hwangryungeehwang-tang Aqueous Extracts on Streptozotocin-induced Diabetic Rat)

  • 유동조;윤경민;강석봉
    • 대한한방내과학회지
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    • 제32권2호
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    • pp.288-300
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    • 2011
  • Objectives : The object of this study was to observe the effects of Hwangryungeehwang-tang (HRGHT), which has traditionally been used in Korean medicine for treating diabetes, by administering aqueous extracts to streptozotocin-induced diabetic rats. Methods : Three different dosages of HRGHT extracts were orally administered once a day for 28 days from 3 weeks after streptozotocin treatment (60 mg/kg, single intraperitoneally administered). 6 groups, of 8 rats per group were used in the present study, selected 3 weeks after streptozotocin treatment based on blood glucose levels and body weights. Changes to the body weights, blood glucose levels, serum low density lipoprotein (LDL), high density lipoprotein (HDL), triglyceride and total cholesterol levels were observed, as well as changes on the pancreatic, hepatic and kidney malondialdehyde (MDA) contents for lipid peroxidation and glutathione contents for antioxidant effects. The results were compared with a potent antioxidant silymarin 100 mg/kg, of which the effects on streptozotocin-induced diabetes and related complications are already confirmed. Results : Significant decrease of blood glucose levels, serum LDL, total cholesterol, triglyceride levels, pancreatic, hepatic and renal MDA contents and significant increase of body weights, serum HDL levels, pancreatic, hepatic and renal GSH contents were detected in HRGHT extracts 100 and 200 mg/kg administered groups as compared to the streptozotocin control group. The lowest dosage of HRGHT treated group, 50 mg/kg, did not show any favorable effects as compared with the streptozotocin control. Conclusions : HRGHT has favorable effects on diabetes and related hyperlipidemia. Therefore, more detailed mechanism studies should be conduced in future with efficacy tests of individual herbal compositions of HRGHT and screening of the biological active compounds in the herbs.