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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.

The study on the cleft lip and/or palate patients who visited Dept. of Orthodontics, Seoul National University Dental Hospital during last 11 years (1988.3-1999.2) (최근 11년간 서울대학교병원 교정과에 내원한 순구개열 환자의 내원 현황에 관한 연구(1988.3 - 1999.2))

  • Yang, Won-Sik;Baek, Seung-Hak
    • The korean journal of orthodontics
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    • v.29 no.4 s.75
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    • pp.467-481
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    • 1999
  • Cleft lip and/or palate is one of the most common congenital craniofacial anomalies. According to previous epidemiologic studies, incidence of cleft lip and/or palate has been increasing nowadays. However, there is no report about epidemiologic study of cleft lip and/or palate patients who visited dept. of orthodontics in Korea. So the purpose of this study was to provide the epidemiological characteristics and important basic clinical data for the diagnosis and the treatment of the cleft lip and/or palate patients. With the orthodontic and cleft charts, diagnostic models and X-ray films from 250 patients with cleft lip and/or palate who visited Dept. of Orthodontics, Seoul National University Dental Hospital during the last 11 years, the authors investigated patient's visiting yew, types of cleft, patient's gender, and Angle's classification of malocclusion, and surgery timing. The results were as follows ; 1. The number of cleft patients who visited Dept. of Orthodontics, SNUDH increased during 1988-1990 and then it declined until 1992. From 1993 to 1996, it showed a stationary trend. After 1997 it showed an overwhelmingly increasing trend. 2. In the cleft type, the ratio of cleft lip cleft lip and alveolus cleft palate : cleft lip and palate was 7.6:19.2:9.6:63.6. In cleft position, unilateral clefts were more than bilateral ones (cleft lip 79:21, cleft lip and alveolus 77:23, cleft lip and palate 75.5:24.5). In cleft side, left clefts were mote than right clefts (cleft lip 53.3:46.7 cleft lip and alveolus 59.5:40.5, cleft lip and palate 59.2:40.8). 3. In gender ratio, males were more than females in cleft lip (57.9:42.1), cleft lip and alveolus (68.8:31.2) and cleft lip and palate (76.1:23.9). But in cleft Palate females were more than males as 41.7: 58.3. 4. In the age groups, 7-12 year group was the most abundant as $52\%$, and then 0-6 year group ($20.4\%$), 13-18 year group ($17.2\%$), more than 18 yew group ($10.4\%$) were followed as descending order. 5. Most of the cleft lip repair surgeries were operated in 0-3 month ($60.3\%$) and 4-6 month ($17.9\%$). 6. The cleft palate repair surgeries were done in 1-2 year ($31.7\%$), 0-1 year ($25.6\%$), 2-3 year ($12.1\%$), more than 5 year ($11.6\%$) as descending order. 7. The lip scar revision surgeries were done before admission at elementary school in $60\%$. (4-6 you ($27.5\%$), 6-8 year ($19.6\%$), more than 10 year ($19.6\%$), 2-4 year ($13.7\%$) as descending order) 8. The rhinoplasties were done before admission at elementary school in $51.7\%$. (0-2 year ($7.1\%$), 2-4 year ($14.3\%$), 4-6 year ($21.4\%$), 6-8 year ($14.3\%$)). 9. The pharyngeal flap were done at 6 Y (72.5 months) after birth on average and there was even distribution of surgery timing. 10. In relationship between Angle's classification of malocclusion and cleft types, Class I was most abundant and Class III, Class II were followed as descending order in cleft lip group. But Class III was most abundant and Class I, Class II were followed as descending order in cleft lip and alveolus group, cleft palate group, and cleft lip and Palate group. The percentage of frequency in Class III malocclusion was overwhelmingly higher in cleft lip and palate group than any other groups. 11. Because the frequency of class III malocclusion was most prevalent in all age groups, anterior crossbite was the most common chief complaint of cleft patients.

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EVALUATION OF CONDYLAR POSITION USING COMPUTED TOMOGRAPH FOLLOWING BILATERAL SAGITTAL SPLIT RAMUS OSTEOTOMY (전산화단층촬영법을 이용한 하악 전돌증 환자의 하악지 시상 골절단술후 하악과두 위치변화 분석)

  • Chol, Kang-Young;Lee, Sang-Han
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.18 no.4
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    • pp.570-593
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    • 1996
  • This study was intended to perform the influence of condyle positional change after surgical correction of skeletal Class III malocclusion after BSSRO in 20 patients(males 9, females 11) using computed tomogram that were taken in centric occlusion before, immediate, and long term after surgery and lateral cephalogram that were taken in centric occlusion before, 7 days within the period intermaxillary fixation, 24hour after removing intermaxillary fixation and long term after surgery. 1. Mean intercondylar distance was $84.45{\pm}4.01mm$ and horizontal long axis of condylar angle was $11.89{\pm}5.19^{\circ}$on right, $11.65{\pm}2.09^{\circ}$on left side and condylar lateral poles were located about 12mm and medial poles about 7mm from reference line(AA') on the axial tomograph. Mean intercondylar distance was $84.43{\pm}3.96mm$ and vertical axis angle of condylar angle was $78.72{\pm}3.43^{\circ}$on right, $78.09{\pm}6.12^{\circ}$on left. 2. No statistical significance was found on the condylar change(T2C-T1C) but it had definitive increasing tendency. There was significant decreasing of the distance between both condylar pole and the AA'(p<0.05) during the long term(TLC-T2C). 3. On the lateral cephalogram, no statistical significance was found between immediate after surgery and 24 hours after the removing of intermaxillary fixation but only the lower incisor tip moved forward about 0.33mm(p<0.05). Considering individual relapse rate, mean relapse rate was 1.2% on L1, 5.0% on B, 2.0% on Pog, 9.1% on Gn, 10.3% on Me(p<0.05). 4. There was statistical significance on the influence of the mandibular set-back to the total mandibular relapse(p<0.05). 5. There was no statistical significance on the influence of the mandibular set-back(T2-T1) to the condylar change(T2C-T1C), the condylar change(T2C-T1C, TLC-T2C) to the mandibular total relapse, the pre-operative condylar position to the condylar change(T2C-T1C, TLC-T2C), the pre-operative mandibular posture to the condylar change(T2C-T1C, TLC-T2C)(p>0.05). 6. The result of multiple regression analysis on the influence of the pre-operative condylar position to the total mandibular relapse revealed that the more increasing of intercondylar distance and condylar vertical axis angle and decreasing of condyalr head long axis angle, the more increasing of mandibular horizontal relapse(L1,B,Pog,Gn,Me) on the right side condyle. The same result was founded in the case of horizontal relapse(L1,Me) on the left side condyle.(p<0.05). 7. The result of multiple regression analysis on the influence of the pre-operative condylar position to the pre-operative mandibular posture revealed that the more increasing of intercondylar distance and condylar vertical axis angle and decreasing of condylar head long axis angle, the more increasing of mandibular vertical length on the right side condyle. and increasing of vertical lengh & prognathism on the left side condyle(p<0.05). 8. The result of simple regression analysis on the influence of the pre-operative mandibular posture to the mandibular total relapse revealed that the more increasing of prognathism, the more increasing of mandibular total relapse in B and the more increasing of over-jet the more increasing of mandibular total relapse(p<0.05). Consequently, surgical mandibular repositioning was not significantly influenced to the change of condylar position with condylar reposition method.

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A Survey on the Perception of the Counterplans of Medical Accident and Dispute of Dental Hygienist (의료사고 및 의료분쟁에 대한 치위생사의 인식도 조사)

  • Oh, Jin-Ho;Kwon, Jeong-Seung;Ahn, Hyoung-Joon;Kang, Jin-Kyu;Choi, Jong-Hoon
    • Journal of Oral Medicine and Pain
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    • v.32 no.1
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    • pp.9-33
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    • 2007
  • In the field of dentistry, there existed relatively few emergency patients or patients who need intensive care and thus had low medical dispute rates. However, these days, there is a general tendency of increased medical disputes. Although many medical disputes are caused by medical accidents of the dentists, because dental assistants are also lawfully involved in practicing dentistry, there is a possibility of medical disputes or medical accidents caused by dental assistants. Therefore, the role of the dental assistants cannot be ignored. This study consists of a survey given to dental hygienists currently working in general hospitals, dental hospitals and private dental clinics. Following is the results of the analysis of 275 respondents' backgrounds, medical disputes rates including patients' complaints, their understanding of medical regulations and their general understanding of overall dental practice and medical disputes. 1. 251 of 274(91.6%) respondents doubted the risk of medical accident and dispute. 2. 81(29.5%) dental hygienist experienced complaint from patients. They have been working in the private dental clinic, the rate of this experience was high. 3. 349 case of 1805(19.3%) the complaints by patients, highest percentage among its category, were those regarding dental fees and poor service. 4. 129 case of 1805(7.1%) patients' complaints, highest percentage among it's subcategory, were those regarding the absence of explanations of precautions or request of agreements before dental treatment. 5. 252 of 267 (94.4%) dental hygienists chart after a scaling treatment. However, only 55(20.7%) dental hygienists chart the fact of explaining the precautions. 6. 6(2.2%) dental hygienists do not inspect patients' medical history, if patients don't mention it. 7. 104 of 274(38.0%) dental hygienists responded to be capable of administering first aid treatment. 8. 115(41.8%) dental hygienists have a first aid kit and equipment. 9. In case of medical dispute, 268(97.8%) dental hygienists respond that, charting plays a big role in resolving the dispute. 10. In case of medical dispute, 272(93.3%) dental hygienists respond that, explanation and agreement before treatment have an important role in settlement of dispute 11. Only 160(58.4%) dental hygienists responded correct answer that the duration of keeping medical records is 10 years. 12. 124(45.3%) respondents thought that it is legal for a dental hygienist to take a panoramic dental X-ray, 71(25.9%) respondents thought that it is legal practice cervical resin treatment by dental hygienist, and 37(13.5%) respondents thought that it is legal extract primary teeth by dental hygienist. 13. 24(18.76%) respondents thought that it doesn't matter to tell patient's state to others 14. 272(99.27%) responded that receiving education for the prevention of medical disputes was needed and of them, 61.0% thought it was urgent. 15. 186(64.2%) has never had classes regarding the prevention of medical disputes while in school and 212(77.4%) has not had the same type of classes after graduating from school. 16. 256(93.4%) responded that there will be even more of an increased number of medical disputes. Among them, 83.3% of respondents though that due to the increased opportunity of acquiring information through the internet and mass media. The study shows that 29.5 percentage of dental hygienists have experienced the medical disputes and complaints and they are lack of recognition of medical regulations and dental hygienist's official duty. So, there is a big potential of the percentage to increase. Therefore, the correct understanding of explaining precautions and requesting agreement before dental treatments and performing them are mandatory. Moreover, classes regarding the prevention and counterplans of medical disputes need to be widely offered.

Halitosis and Related Factors among Rural Residents (농촌지역 주민들의 구취실태와 유발요인)

  • Lee, Young-Ok;Hong, Jung-Pyo;Lee, Tae-Yong
    • Journal of Oral Medicine and Pain
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    • v.32 no.2
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    • pp.157-175
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    • 2007
  • This study was conducted through an interview process in which questionnaires were administered to 293 people. The questionnaires related to the behaviors of oral hygiene care, and disease history related to halitosis, and status of halitosis, halitosis measurement, oral examination, and caries activity tests such as the snyder test, Salivary flow rate test, and Salivary buffering capacity test. Our sample was taken from 293 rural residents within the period from 4th to 21st of January 2006. This was done in order to provide basic data to prepare both policies of halitosis prevention and a device to efficiently measure halitosis status and investigate the factors related therein. The major findings of this study results are as follows: 1. As for frequency of tooth brushing, twice a day occupied the greatest portion at 46.1 % Women exceeded men in frequency of tooth brushing. Tongue brushing everyday produced a 25.6 % result among subjects and The use of auxiliary oral hygiene devices occupied 9.2 %. 2. As for degree of usual self-awareness of halitosis: 62.5 %. This result also demonstrate that the severest time of self-awareness in regards to halitosis is wake up time in the morning. The time period produced the highest portion of 72.7 % in times of self-awareness. In terms of the area in which halitosis was observed, gum resulted in 23.0 %. As for types of halitosis, fetid smell was the most frequent at 37.2 %. 3. As for the result of halitosis measurement, values of OG less than 50 ppm occupied 54.3 % and $50{\sim}100ppm$ occupied 41.6 %. As for $NH_3$ values, $20{\sim}60ppm$ showed the highest value range of 52.6 %. 4. As for OG per disease history related to halitosis, values of OG were significantly high in the ranges of $50{\sim}100ppm$ within family history groups of food impaction by dental caries, diabetes mellitus and halitosis. As for values of $NH_3$, there showed a significant difference in respiratory system disease groups. 5 Value range of OG per ordinary halitosis self-awareness degree: values ranging less than 50 ppm were recorded at 55.9 % from the group realizing not aware of smell. 57.5 % from groups only realizing sometimes, while values range of $50{\sim}100ppm$ were recorded at 52.0 % from groups always aware of smell. 63.6 % from groups always strongly aware of smell. Meanwhile as for the values ranges of $NH_3$, $20{\sim}60ppm$. they occupied high portions for all groups of exams. 6. Values of OG per oral examination: the more pulp-exposed teeth and food impaction and the higher the tongue plaque index, values of OG increased within the range of $50{\sim}100ppm$. As for values of $NH_3$, the more prosthetic teeth and the higher the tongue plaque index, this value increased significantly, and the values increased up to no less than 60 ppm for groups of mandibular partial denture. 7. Within the realm of caries activity test: as for the Snyder test, high activity was highest by 43.0 % wherewith the higher the activity of acidogenic bacteria the higher the OG values. As for the salivary flow rate test, the number of cases below 8.0 ml showed the highest tendency by 62.5 %. The larger the salivary flow rate the more decreased OG values distribution. As for the salivary buffering capacity test, $6{\sim}10$ drops of 0.1N lactic acid showed the overwhelming trend by 58.7 % whereby the higher the salivary buffering capacity the greater distribution occupancy ratio of OG values below 50 ppm which is scentless to on ordinary person. 8. As for the correlation between oral environment and halitosis, OG showed the positive correlation with pulp exposed teeth, filled teeth, present teeth, tongue plaque index, and food impaction, while the negative correlation with salivary flow rate and prosthetic teeth. $NH_3$ showed a positive correlation with prosthetic teeth and frequency of tooth brushing, while decayed teeth was negative correlation. 9. As for the multiple regression analysis result, there have been selected female, pulp exposed teeth, prosthetic teeth, food impaction, salivary flow rate, tongue plaque index and severe activities in the Snyder test as factors affecting OG wherein explanatory power on it was 45.1 %. There have been selected females, pulp exposed teeth, tongue plaque index, and prosthetic teeth as factors affecting on $NH_3$ wherein explanatory power on it was 6.6 %. With the aforementioned results in mind, the status of halitosis among rural residents is considered to bare a close relation with oral environments and other factors related to halitosis such as the Snyder test from caries activity test, and salivary flow rate test. For the prevention of halitosis of residents in rural areas, we have to focus on correct tooth brushing methods and tongue brushing, with using auxiliary oral hygiene devices to remove fur of tongue plaque and food impaction. Also, when the cause and ingredients of halitosis are diverse and complex, in order to analyze exactly the factors of individual halitosis development, we need continuous and systematic study in order to provide rural residents with programs of oral hygiene education and encourage the use of dental hygienists in public health centers.