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The Effects of Storage of Human Saliva on DNA Isolation and Stability (인체타액의 보관이 DNA 분리와 안정도에 미치는 영향)

  • Kim, Yong-Woo;Kim, Young-Ku
    • Journal of Oral Medicine and Pain
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    • v.31 no.1
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    • pp.1-16
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    • 2006
  • The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of micromethodologies and increasing availability of immunological and molecular biological reagents. The technological advances led to consider the diagnostic use of saliva for an array of analytes and DNA source. The purpose of the present study was to compare DNA from saliva with those from blood and buccal swab, to evaluate diagnostic and forensic application of saliva, to investigate the changes of genomic DNA in saliva according to the storage temperature and period of saliva samples, and to evaluate the integrity of the DNA from saliva stored under various storage conditions by PCR analysis. Peripheral venous blood, unstimulated whole saliva, stimulated whole saliva, and buccal swab were obtained from healthy 10 subjects (mean age: $29.9{\pm}9.8$ years) and genomic DNA was extracted using commercial kit. For the study of effects of various storage conditions on genomic DNA from saliva, stimulated whole saliva were obtained from healthy 20 subjects (mean age: $32.3{\pm}6.6$ years). After making aliquots from fresh saliva, they were stored at room temperature, $4^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$. Saliva samples after lyophilization and dry-out procedure were stored at room temperature. After 1, 3, and 5 months, the same experiment was performed to investigate the changes in genomic DNA in saliva samples. In case of saliva aliquots stored at room temperature and dry-out samples, the results in 2 weeks were also included. Integrity of DNA from saliva stored under various storage conditions was also evaluated by PCR amplification analysis of $\beta$-globin gene fragments (989-bp). The results were as follows: 1. Concentration of genomic DNA extracted from saliva was lower than that from blood (p<0.05), but there were no significant differences among various types of saliva samples. Purities of genomic DNA extracted from stimulated whole saliva and lyophilized one were significantly higher than that from blood (p<0.05). Purity of genomic DNA extracted from buccal swab was lower than those from various types of saliva samples (p<0.05). 2. Concentration of genomic DNA from saliva stored at room temperature showed gradual reduction after 1 month, and decreased significantly in 3 and 5 months (p<0.05, p<0.01, respectively). Purities of DNA from saliva stored for 3 and 5 months showed significant differences with those of fresh saliva and stored saliva for 1 month (p<0.05). 3. In the case of saliva stored at $4^{\circ}C$ and $-20^{\circ}C$, there were no significant changes of concentration of genomic DNA in 3 months. Concentration of DNA decreased significantly in 5 months (p<0.05). 4. There were no significant differences of concentration of genomic DNA from saliva stored at $-70^{\circ}C$ and from lyophilized one according to storage period. Concentration of DNA showed decreasing tendency in 5 months. 5. Concentration of genomic DNA immediately extracted from saliva dried on Petri dish were 60% compared with that of fresh saliva. Concentration of DNA from saliva stored at room temperature after dry-out showed rapid reduction within 2 weeks (p<0.05). 6. Amplification of $\beta$-globin gene using PCR was successful in all lyophilized saliva stored for 5 months. At the time of 1 month, $\beta$-globin gene was successfully amplified in all saliva samples stored at $-20^{\circ}C$ and $-70^{\circ}C$, and in some saliva samples stored at $4^{\circ}C$. $\beta$-globin gene was failed to amplify in saliva stored at room temperature and dry-out saliva.

A Study on the Improvement of Flexible Working Hours (탄력적 근로시간제 개선에 대한 연구)

  • Kwon, Yong-man
    • Journal of Venture Innovation
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    • v.5 no.3
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    • pp.57-70
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    • 2022
  • In modern industrial capitalism, the relationship between the provision of work and the receipt of wages has become an important principle governing society. According to the labor contract, the wages provided by entrusting the right to dispose of one's labor to the employer are directly compensated, and human life should be guaranteed and reproduced with proper rest. The establishment of labor relations under free contracts represents a problem in protecting workers, and accordingly, the maximum of working hours is set as a minimum right for workers, and the standard for minimum rest is set and assigned. The reduction of working hours is very important in terms of the quality of life of workers, but it is also an important issue in efficient corporate activities. As of 2020, Korea has 1,908 hours of annual working hours, the third lowest among OECD 37 countries in the happiness index surveyed by the Sustainable Development Solution Network(SDSN), an agency under the United Nations. Accordingly, the necessity of reducing working hours has been recognized, and the maximum working hours per week has been limited to 52 hours since 2018. In this situation, various working hours are legally excluded as a way to maintain the company's value-added creation and meet the diverse needs of workers, and Korea's Labor Standards Act restricts flexible working hours within three months, flexible working hours exceeding three months, selective working hours, and extended working hours. However, in the discussion on the application of the revised flexible working hours system in 2021 and the expansion of the settlement unit period recently discussed, there is a problem with the flexible working hours system, which needs to be improved. Therefore, this paper aims to examine the problems of the flexible working hours system and improvement measures. The flexible working hours system is a system that does not violate working hours even if the legal working hours are exceeded on a specific day or week according to a predetermined standard, and does not have to pay additional wages for excessive overtime work. It is mainly useful as a form of shift work in manufacturing, sales service, continuous business or electricity, gas, water, and transportation for long-term operations. It is also used as a way to shorten working hours, such as expanding holidays through short working days. However, if the settlement unit period is expanded, it is disadvantageous to workers as the additional wages that workers can receive will not be received. Therefore, First, in order to expand the settlement unit period currently under discussion, additional wages should be paid for the period expanded from the current standard. Second, it is necessary to improve the application of the flexible working hours system to individual workers to have sufficient consultation with individual workers in a written agreement with the worker representative, Third, clarify the allowable time for extended work during the settlement unit period, and Fourth, limit the daily working hours or apply to continuous rest. In addition, since the written agreement of the worker representative is an important issue in the application of the flexible working hours system, it is necessary to secure the representation of the worker representative.