• Title/Summary/Keyword: Biological sample

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Rapid Detection of Clostridium tetani by Recombinase Polymerase Amplification Using an Exo Probe

  • Guo, Mingjing;Feng, Pan;Zhang, Liqun;Feng, Chunfeng;Fu, Jie;Pu, Xiaoyun;Liu, Fei
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.91-98
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    • 2022
  • Tetanus is a potentially fatal public health illness resulted from the neurotoxins generated by Clostridium tetani. C. tetani is not easily culturable and culturing the relevant bacteria from infected wounds has rarely been useful in diagnosis; PCR-based assays can only be conducted at highly sophisticated laboratories. Therefore, a real-time recombinase polymerase amplification assay (Exo-RPA) was constructed to identify the fragments of the neurotoxin gene of C. tetani. Primers and the exo probe targeting the conserved region were designed, and the resulting amplicons could be detected in less than 20 min, with a detection limit of 20 copies/reaction. The RPA assay displayed good selectivity, and there were no cross-reactions with other infectious bacteria common in penetrating wounds. Tests of target-spiked serum and pus extract revealed that RPA is robust to interfering factors and has great potential for further development for biological sample analysis. This method has been confirmed to be reliable for discriminating between toxic and nontoxic C. tetani strains. The RPA assay dramatically improves the diagnostic efficacy with simplified device architecture and is a promising alternative to real-time PCR for tetanus detection.

Monitoring the presence of wild boar and land mammals using environmental DNA metabarcoding - Case study in Yangpyeong-gun, Gyeonggi-do - (환경 DNA 메타바코딩을 활용한 멧돼지 및 육상 포유류 출현 모니터링 - 경기도 양평군 일대를 중심으로 -)

  • Kim, Yong-Hwan;Han, Youn-Ha;Park, Ji-Yun;Kim, Ho Gul;Cho, Soo-Hyun;Song, Young-Keun
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.24 no.6
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    • pp.133-144
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    • 2021
  • This study aims to estimate location of land mammals habitat by analyzing spatial data and investigate how to apply environmental DNA monitoring methodology to lotic system in Yangpyeong-gun, Gyeonggi-do. Environmental DNA sampling points are selected through spatial analysis with QGIS open source program by overlaying Kernel density of wild boar(Sus scrofa), elevation, slope and land-cover map, and 81 samples are collected. After 240 mL of water was filtered in each sample, metabarcoding technique using MiMammal universal primer was applied in order to get a whole list of mammal species whose DNA particles contained in filtered water. 8 and 22 samples showed DNA of wild boar and water deer, respectively. DNA of raccoon dog, Eurasian otter, and Siberian weasel are also detected through metabarcoding analysis. This study is valuable that conducted in outdoor lotic system. The study suggests a new wildlife monitoring methodology integrating overlayed geographic data and environmental DNA.

Psychosocial Risks Assessment in Cryopreservation Laboratories

  • Fernandes, Ana;Figueiredo, Margarida;Ribeiro, Jorge;Neves, Jose;Vicente, Henrique
    • Safety and Health at Work
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    • v.11 no.4
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    • pp.431-442
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    • 2020
  • Background: Psychosocial risks are increasingly a type of risk analyzed in organizations beyond chemical, physical, and biological risks. To this type of risk, a greater attention has been given following the update of ISO 9001: 2015, more precisely the requirement 7.1.4 for the process operation environment. The update of this normative reference was intended to approximate OHSAS 18001: 2007 reference updated in 2018 with the publication of ISO 45001. Thus, the organizations are increasingly committed to achieving and demonstrating good occupational health and safety performance. Methods: The aim of this study was to characterize the psychosocial risks in a cryopreservation laboratory and to develop a predictive model for psychosocial risk management. The methodology followed to collect the information was the inquiry by questionnaire that was applied to a sample comprising 200 employees. Results: The results show that most of the respondents are aware of the psychosocial risks, identifying interpersonal relationships and emotional feelings as the main factors that lead to this type of risks. Furthermore, terms such as lack of resources, working hours, lab equipment, stress, and precariousness show strong correlation with psychosocial risks. The model presented in this study, based on artificial neural networks, exhibited good performance in the prediction of the psychosocial risks. Conclusion: This work presents the development of an intelligent system that allows identifying the weaknesses of the organization and contributing to the enhancement of the psychosocial risks management.

A Sensitive, Efficient, and Cost-Effective Method to Determine Rotigotine in Rat Plasma Using Liquid-Liquid Extraction (LLE) and LC-MRM

  • Kim, Ji Seong;Jang, Yong Jin;Kim, Jin Hee;Kim, Jin Hwan;Seo, Jae Hee;Park, Il-Ho;Kang, Myung Joo;Choi, Yong Seok
    • Mass Spectrometry Letters
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    • v.13 no.4
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    • pp.146-151
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    • 2022
  • Rotigotine (RTG) is a non-ergot dopamine agonist used to manage the early stage of Parkinson's disease (PD) as transdermal patch. However, the poor medication compliance of PD patients and skin issues related with repeated applications of RTG patches lead to the search for alternative formulations and it also requires appropriate analytical methods for their in vivo evaluation. Thus, here, a sensitive, efficient, and cost-effective method to determine RTG in rat plasma using liquid-liquid extraction (LLE) and multiple reaction monitoring was developed. The use of 20 µL of rat plasma for sample treatment, 8-OH-DPAT as the internal standard, and methyl tert-butyl ether as the LLE solvent in the present method gives it advantages over previous methods for the analysis of RTG in biological samples. The good analytical performance of the developed method was confirmed in specificity, linearity (the coefficient of determination ≥0.999 within 0.1-100 ng/mL), sensitivity (the lower limit of quantitation at 0.1 ng/mL), accuracy (81.00-115.05%), precision (≤10.75%), and recovery (81.00-104.48%) by following the FDA guidelines. Finally, the applicability test of the validated method to the in vivo evaluation of a RTG formulation showed that the present method is the only method which can be accurately applied to that longer than 24 hours, critical for the development of formulations with reduced dosing frequencies. Therefore, the present method could contribute to the development of new RTG formulations helpful to people suffering from PD.

Factors Influencing the Intake of Convenience Foods among Korean Adolescents: Based on the bio-psycho-social model (한국 청소년의 편의식품 섭취 경험의 영향요인 : 생물-심리-사회 모델을 바탕으로)

  • Moon-Hee Kang;Soon-Ok Kim;Yeon-Hee Jeong
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.3
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    • pp.547-559
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    • 2023
  • The purpose of this study was to identify the influencing factors of Korean adolescents' consumption of convenience foods based on a bio-psycho-social model. Data were obtained from the 15th (2019) Korea Youth Risk Behavior Survey, which involved a nationwide representative sample of 57,303 middle- and high-school students. As a result of the study, gender, grade, and atopic dermatitis were biological factors of convenience food intake in adolescents. The psychological factors were perceived stress level, experience of depression, suicidal ideation, subjective sleep sufficiency, perceived health status, physical activity, and current smoking and drinking. And nutrition and meal education, breakfast frequency, subjective academic achievement, subjective economic status, and family living together were sociological factors that affected adolescents' convenience food intake experience. Therefore, it is necessary to provide effective nutrition education programs and provide emotional support to manage healthy eating habits of adolescents. In addition, family members of adolescents should be included in education.

Discrimination of biological and artificial nicotine in e-liquid

  • Hyoung-Joon Park;Heesung Moon;Min Kyoung Lee;Min Soo Kim;Seok Heo;Chang-Yong Yoon;Sunyoung Baek
    • Analytical Science and Technology
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    • v.36 no.1
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    • pp.22-31
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    • 2023
  • As the use of e-liquid cigarettes is rapidly increasing worldwide, it multiplies the potential risk undisclosed to the health of non- and smokers. To reduce the hazard, each country has its own set of regulations for controlling e-liquids. In Korea, the narrow definition of tobacco makes it difficult and have been steadily occurring tax evasion exploiting the difference in natural and artificial nicotine. Therefore, it is very important to distinguish source of nicotine for their regulation. To find biochemical discriminant markers, this study established analysis methods based on high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) and high-performance liquid chromatography coupled with triple Quadrupole mass spectrometry (HPLC-MS/MS) for nicotine enantiomers and tobacco alkaloids targeted using the difference in pathways of nicotine biosynthesis and chemical synthesis. The method was validated by experimenting linearity (R2 > 0.999), recovery (80.99-108.41 %), accuracy (94.11-109.73 %) and precision (0.04-8.27 %). Then, the results for discrimination of the nicotine obtained from analysis of 65 commercial e-liquid products available in Korean market was evaluated. The method successfully applied to the e-liquids and one sample labelled 'synthetic nicotine' for tax exemption was found to contain a natural nicotine product. This method can be used to determine whether an e-liquid product uses natural or artificial nicotine and monitor non-taxable e-liquid products. The method is more scientific than the existing one, which relies only on field evidence.

Determination of diallyl disulfide in garlic by reversed-phase high performance liquid chromatography (역상 고성능 액체 크로마토그래피를 이용한 마늘에서 diallyl disulfide의 분석)

  • Wan, Xiaolong;Polyakova, Yulia;Row, Kyung Ho
    • Analytical Science and Technology
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    • v.20 no.5
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    • pp.442-447
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    • 2007
  • The properties of garlic (Allium sativum L.) are attributed to organosulfur compounds. In this paper, an analytical technique with a rapid and simple sample preparation procedures for determination of diallyl disulfide (DADS) in garlic was reported. The DADS was simply extracted with various solvents (methanol, benzene or tetrahydrofuran) from garlic and prepared for HPLC analysis. From the results, the methanol was select as an optimal extraction solvent. The mobile phase was composed from methanol and water, and the gradient elution mode was applied. 0.61 mg of DADS per g garlic powder can be extracted with methanol. This work offers some advantages over the currently accepted techniques and would be useful for chemical and biological studies of garlic and its products.

Analysis of Endophytic Bacterial Communities and Investigation of Core Taxa in Apple Trees

  • Yejin Lee;Gyeongjun Cho;Da-Ran Kim;Youn-Sig Kwak
    • The Plant Pathology Journal
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    • v.39 no.4
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    • pp.397-408
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    • 2023
  • Fire blight disease, caused by Erwinia amylovora, is a devastating affliction in apple cultivation worldwide. Chemical pesticides have exhibited limited effectiveness in controlling the disease, and biological control options for treating fruit trees are limited. Therefore, a relatively large-scale survey is necessary to develop microbial agents for apple trees. Here we collected healthy apple trees from across the country to identify common and core bacterial taxa. We analyzed the endophytic bacterial communities in leaves and twigs and discovered that the twig bacterial communities were more conserved than those in the leaves, regardless of the origin of the sample. This finding indicates that specific endophytic taxa are consistently present in healthy apple trees and may be involved in vital functions such as disease prevention and growth. Furthermore, we compared the community metabolite pathway expression rates of these endophyte communities with those of E. amylovora infected apple trees and discovered that the endophyte communities in healthy apple trees not only had similar community structures but also similar metabolite pathway expression rates. Additionally, Pseudomonas and Methylobacterium-Methylorobrum were the dominant taxa in all healthy apple trees. Our findings provide valuable insights into the potential roles of endophytes in healthy apple trees and inform the development of strategies for enhancing apple growth and resilience. Moreover, the similarity in cluster structure and pathway analysis between healthy orchards was mutually reinforcing, demonstrating the power of microbiome analysis as a tool for identifying factors that contribute to plant health.

Effect of Demographic Factors, Radiation Knowledge Level, Radiation Awareness on Radiation Benefit by Hierarchical Regression Analysis Model (위계적 회귀분석 모형에 의한 인구학적 요인, 방사선 지식수준, 방사선 인식도가 방사선 이익성에 미치는 영향)

  • Myeong-Hoon Ji;Youl-Hun Seoung
    • Journal of radiological science and technology
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    • v.46 no.5
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    • pp.435-444
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    • 2023
  • The purpose of this study was to analyze the factors that demographic factors, radiation knowledge level, and radiation awareness could be affecting the benefits of radiation. From July 2022 to July 2023, after receiving consent to participate by using the link of Naver through Social Network Service (SNS) for the general public, 312 people were surveyed by self-registration method without collecting personal information. The questionnaire consisted of a total of 25 questions following demographic factors (5 questions including age group by life cycle, sex, monthly household income, residence), radiation knowledge level (8 questions including basic physical, biological effects, radiation protection technology), radiation awareness (12 questions including risk, management, benefit). Independent sample T-test and ANOVA tests were performed for significant differences in the average radiation awareness between variables, and hierarchical regression was performed to identify influencing factors on radiation benefits. As a result, the benefit of radiation was significantly high among the radiation awareness, but the awareness of the danger of radiation was insufficient to the level of recognizing it as safe. Men had significantly higher awareness of radiation management and benefits than women, and the awareness of radiation management was significantly higher in the middle class with a monthly household income of 4.31 million won or more. The higher the knowledge level of radiation, the higher the awareness of the benefits of radiation. The factors that had a positive effect on radiation benefits were the high level of radiation knowledge and awareness of radiation management.

LC-MS/MS-based Quantification of Ten Neurotransmitters in Rat Limbic System and Serum: Application to Chronic Unpredictable Mild Stress-Induced Depression Rats

  • Mingyan Ma;Qiangxiang Chen;Wen Cao;Yubo Zhou;Aijuan Yan;Yanru Zhu
    • Mass Spectrometry Letters
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    • v.14 no.3
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    • pp.91-103
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    • 2023
  • As one of the most common mood disorders, numerous studies have shown depression is the main risk factor for non-suicidal self-harm. The pathogenesis of depression is complex, and a comprehensive and rapid measurement of monoamine neurotransmitters and their metabolites will be very helpful in understanding the pathogenesis of depression. Therefore, a rapid and sensitive underivatized liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous monitoring of the levels of ten neurotransmitters and their metabolites in rat serum and limbic system and successfully applied to quantify the changes of neurotransmitter levels in chronic unpredictable mild stress-induced rats. The analytes studied were mainly involved in tyrosine metabolism, tryptophan metabolism, and glutamate cycling pathways, which are important in the pathogenesis of depression. It had been verified the method was sensitive and effective, with satisfactory linearity, and met the requirements of biological sample determination. Levels of neurotransmitters in rat serum, hippocampus, amygdala, prefrontal cortex, striatum, and hypothalamus were determined via the method. The results showed serotonin, dopamine, norepinephrine, and their metabolites were decreased, glutamine was increased, and glutamate was disturbed in chronic unpredictable mild stress-induced depression rats. This method provides a new approach to studying the pathogenesis of depression and other neurological disorders.