DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.
Journal of the korean academy of Pediatric Dentistry
/
v.38
no.1
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pp.1-8
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2011
The objective of this study was to determine the effectiveness of topical 5% EMLA cream versus 20% Benzocaine gel in reducing pain from intra oral needle insertion alone as well as injection of anesthetic. The 2 topical anesthetics were tested against each other bilaterally using a randomized, controlled, single blinded, split mouth design. Phase I was conducted to find out the rapidity of onset action of the two agents on anterior/posterior vestibules and anterior/posterior palatal mucosa. Phase II was conducted to evaluate the efficacy of the two topical anaesthetic agents in reducing the pain of intraoral injections. The agents were left in anterior/posterior vestibules and anterior/posterior palatal mucosa for the amount of time recorded in phase I. Subjects recorded pain on a 100-mm modified visual analog scale(VAS). A pulse oximeter was used to recorded the preoperative and postoperative pulse rates. In phase I of the study, two topical agents showed the longer onset of action at anterior part and vestibules than posterior part and palatal mucosa. EMLA cream showed the rapidest onset of action compared to benzocaine gel except on anterior palatal mucosa. In phase II of the study, the VAS grading of the pain for anesthetic administration showed EMLA cream was significantly(P<.05) better in elimination or reducing the pain on the anterior/posterior palatal mucosa. In conclusion, EMLA cream showed the rapidest onset of action compared to benzocaine gel except on anterior palatal mucosa. EMLA cream was found to be superior to Benzocaine gel with regards to pain reduction for anesthetic administration especially on anterior and posterior palatal mucosa.
Occupational health services in Korea have been operated as dual types : one is operated by occupational health care manager and the other is health care agency without their own personnel. The performance of occupational health service should be different due to the variety of characteristics of health care manager and workplace, qualification of health care manager. This study is to analyze performance of occupational health care services with a particular consideration of job performance shape and efficiency, based on comparing those two types of health care management to show on the basic data for the settlement of more qualitative health care management system at workplace. For this study, total 391 places in Seoul and Inchon city area ; 154 places (39.4%) managed by designated health care manager and 237 places (60.6%) by the agency with their commission are selected as research samples. Tools for data collection are questionnares that have been investigated during the period of 20 September 1993-20 December 1993. Those data are compared with percentiles, mean, standard deviation and B/C ratio using SPSS PC program. Conclusions observed from the tests and each comparison could be summerized as follows : 1. Occupational health care have been accomplished at workplaces with designated people than with agencies people, and coverage rate of the occupational health care services has differences, due to management types. The reason of these results is due to visit only one or two times monthly by the agencies, while their own health care manager obsess, at the workplaces all the times. 2. Most of the expense for environmental control of all health care services expenditures shows that there is almost no fundamental improvement because more expenses are needed for procuring personal protective equipment and measuring work environment instead of environmental improvement. 3. It is investigated how much the cost of occupational health care services needs per worker, and calculated how much the cost needs per service hour per worker. The results from this show that the cost of occupational health services at workplaces with their own managers used less than the cost of health care agencies, eventually the former gives better services with less cost than the latter. 4. Benefit/Cost ratio is also produced by total benefit/total cost. The result from the above way reads 4.57 as a whole, while their own manager having workplaces reads 4.82 and the agencies do l.56. Even if their own manager performing workplaces spent more cost, this system produces more benefit than the agencies management. 5. The B/C ratio for medical organization such as local clinic, health care center and pharmacy shows more than or equal to at the workplaces controlled by the agencies. It is inferred that benefit would be much less than the cost used, with so being inefficient. 6. It is assumed that the efficiency ratio of health education is equal to reduction rate of workers medical organization visit. Estimated reduction rate 5%, 10%, 15%, show that the efficiency ratio of health education have an effect on producing benefits. It is estimated that more benefit can be produced if more qualitative education will be provided for enhancing health care efficiency. 7. Results of this study cannot be generalized because there are large scale of deviation in case of workplaces with less than 300 full time workers, but B/C ratio reads 2.69 as a whole and 3.25 at workplaces with their own health care manager are higher than 1.63 at the workplaces manged by the agencies. Finally, all the benefit concerning health care services could not be quantified, measured and shown on the value of money. This is a reason that a considerable part of benefits are so underestimated. This is also thought that measurement tools should be developed for measuring benefits of health care services with a comprehensive quantification. in the future. It is also expected that efficiency of occupational health care services should be investigated using cost-effectiveness analysis.
Background and Methods: To study the effects of corticosteroid (CS) on the parquat (PQ) induced lung injury, serial cellular analyses of bronchoalveolar lavage (BAL) fluid were done with simultaneous histopathologic examination after intraperitoneal injection of PQ on the rats. The sacrificed animals were divided into three groups; control group, PQ group received intraperitoneal injection of 20 mg/kg of PQ, and CS group received daily injection of Methylprednisolone sodium succinate (20 mg/kg) in addition to PQ. Results: 1) Cellular analyses of BAL fluid: The total cell count in the BAL fluid were increased gradually from 6 hours after PQ administration (p<0.05), and was decreased at 3 days after (p<0.05). These changes were mainly due to the effects of PQ on the neutrophil influx (p<0.05). But, the number of macrophage and the percentage of lymphocyte in total cells showed little changes. The CS administration showed the suppression of neutrophil influx in the BAL fluid (p<0.05), but could not show any significant effect on the number of macrophage and lymphocyte. 2) Histopathologic examination: In the PQ group, inflammatory changes especially with prominant neutrophil infiltration were gradually progressed over time. Those changes were found in both alveolar space and interstitium with resultant alveolar structural changes, but subsided from 3 days after. CS suppressed inflammatory changes in the alvolar space and interstitium, especially with decreased infiltration of neutrophil. Conclusion: CS suppressed neutrophil infiltration in the acute lung injury induced by PQ, those findings were ascertained by serial cellular analyses of BAL fluid and histopathologic examination.
Background : Complicated exudative pleural fluid collections have traditionally been treated by either closed tube thoracostomy drainage or by open surgical drainage. Complete drainage is important in order to control pleural sepsis, restore pulmonary function, and entrapment. Recently intracavitary fibrinolytic therapy has been advocated as a method to facillitate drainage of complicated exudative pleural effusion and to allow enzymatic debridemant of the restrictive fibrinous sheets covering the pleural surface. The purpose of this study is to prospectively evaluate the effects of image-guided catheter drainage with high dose urokinase(UK) instillation in the treatment of complicated pleural effusions. Patients : Twenty complicated pleural effusion patients that poorly respond to image-guided drainage were allocated to receive UK. There were 8 pneumonia and 12 tuberculosis. Methods : Drugs were diluted in 250 mL normal saline and were infused intrapleurally through the chest tube or pig-tail catheter in a daily dose of 250,000 IU of UK. Response was assessed by clinical outcome, fluid drainage, chest radiography, pleural ultrasound and/or computed tomography. Results : The mean UK instillation time was $1.63{\pm}0.10$. The mean volume drained UK instillation was $381.3{\pm}314.4\;mL$, and post-UK was $321.6{\pm}489.5\;mL$. The follow up duration after UK therapy was mean $212.9{\pm}194.5$ days. We had successful results in 19 cases (95.0%). There were 12 pleural thickenings (60.0%), 2 markedly decreased effusions (10.0%) and 5 cases of no thickening or effusion. There was recurrence after treatment in only one patient(5%) with complicated pleural effusiondue to tuberculosis. Conclusions : Image-guided drainage with high dose UK instillation (250,000 U/day) in complicated pleural effusion is a safe and more effective method than closed thoracostomy drainage. And this management, in turn, can obviate surgery in most cases.
Kim, Doh-Hyung;Bae, Gang-U;Yong, Wha-Shim;Choi, Eun-Kyung;Kim, Youn-Seup;Park, Jae-Seuk;Jee, Young-Koo;Lee, Kye-Young
Tuberculosis and Respiratory Diseases
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v.53
no.3
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pp.275-284
/
2002
Background : Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. Methods : A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. Results : Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. Conclusion : Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene, Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.
Although the decline in fertility rate is generally observed along the history of economic development throughout the world, the continuing decline hitting below the replacement level in Korea over the recent years gathered serious social concerns on the ground that it accelerates the process of population aging. The total fertility rate in Koreareached 2.08 in 1983, and gradually fell to the levels of 1.08 in 2005 and 1.26 in 2007. The policy debate over the role of the government has been focused mainly on the level of theoretical discussion without substantial basis on firm empirical evidence and the determinants of fertility. The objective of the paper is to empirically investigate the fertility effect of the female wage, which is understood as one of the most important determinants of fertility in Koreasince 1980 focusing on one aspect of fertility, namely birth spacing. Using the Korean National Fertility Survey conducted in 2006, I estimate a duration model of first and second births taking into account individual heterogeneity, which turned out to be an important factor to control for. Compared with previous studies in the literature on the Korean fertility, the study has an advantage of using the complete pregnancy history of women in a more representative sample. Unlike the previous studies, the analysis also deals with the endogeneity of marriage by treating a certain age, rather than age at marriage, as the time in which a woman becomes exposed to the risk of pregnancy. The study shares the common problem in the literature on birth spacing of lacking relevant wage information for respondents in a retrospective survey. I estimate the wage series as a function of the basic characteristics using the annual Wage Structure Survey from 1980 to 2005, which is considered as a nationally representative sample for wage information of employees. The results suggest that the increase in female wage by 10 percent leads to a decrease in second birth hazard by 0.56~0.92 percentage points and that the increase in spouse's wage by the equal amount is accompanied by the increase in second birth hazard by 0.36~1.13 percentage points. These estimates are more precisely estimated and of smaller magnitude than those presented by the previous studies. The results are robust to the different specifications of the wage equation. The simulation analysis based on the predicted values shows that about 17% of the change in the second birth hazard over the period 1980 to 2005 was due to the change in the female wage. Although there is some limitation in data, the results can be viewed as one estimate of the role of female wage on the recent fertility decline in Korea. The question raised by the paper is not a normative one of whether a government should promote childbearing but a positive one thatexplains fertility decline. Therefore, if there is a wide consensus on promoting childbearing, the finding suggests that the policies designed to reduce the opportunity cost of women in the labor market would be effective. The recent movement of implementing a wide range of family-friendly policies including child care support, maternity leave, parental leave and tax benefit in developed countries should be understood in this context.
Kim, Ok-Bae;Kim, Jin-Hee;Cha, Soon-Do;Choi, Tae-Jin;Ye, Ji-Won
Radiation Oncology Journal
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v.22
no.2
/
pp.124-129
/
2004
Purpose : To evaluate the clinical findings, prognosis and treatment strategy of patients with neuroendocrine carcinoma of cervix. Materials and Methods: Thirteen patients with neuroendocrine carcinoma of cervix were included in this study, as confirmed histologically and immunohistochemically at the Dongsan Medical Center, Keimyung University, between May 1944 and October 2001. The mean age of patients was 56 years with a range of 32 to 78 years of age. According to the FIGO staging system, there were 5 patients with stage IB carcinoma, 5 patients with IIA, and 3 patients with stage IIB. Four patients underwent radical hysterectomy with pelvic lymphadenectomy, 3 of these patients also received postoperative radiotherapy, and one patient underwent extrafascial hysterectomy after radiotherapy. Primary radiotherapy was done in 9 patients, and 3 were irradiated postoperatively. Nine patients received chemotherapy, 7 received neoadjuvant and 2 received concurrent chemoradiotherapy. The radical purpose of radiotherapy consisted of external irradiation to the whole pelvis (4,500 $\~$5,400 cGy) and intracavitary irradiation (3,000 $\~$ 3,500 cGy). Results : The mean follow up duration was 36 months with a range of 3 to 104 months. The overall 5-year survival rate was 61.5$\%$ and the 5-year survival rates for stage IB, IIA, IIB were 60.0$\%$, 60.0$\%$, and 66.7$\%$ respectively (p=0.99). Eight patients are still alive without disease, and all of the 5 patients with recurrence are dead due to distant metastasis. Conclusion : Neuroendocrine carcinoma of cervix is highly aggressive, with early lymphatic dissemination and a high rate of distant recurrence. Therefore, an aggressive therapeutic strategy Is required to obtain pelvic and distant disease control. Multimodal therapy should be considered at the time of Initial diagnosis.
Lee Sang-wook;Kim Jae-Seung;Im Ki Chun;Ryu Jin Sook;Lee Hee Kwan;Kim Jong Hoon;Ahn Seung Do;Shin Seong Soo;Yoon Sang Min;Song Siyeol;Park Jin-hong;Moon Dae Hyuk;Choi Eun Kyung
Radiation Oncology Journal
/
v.22
no.2
/
pp.98-105
/
2004
Purpose : To evaluate whether positron omission tomography (PET) with 2-[F-18]fluoro-2-deoxy-D-giucose(FDG) can be used to predict of early response to definitive aim radlotherapy (RT) in squamous cell carcinoma of the head and neck using response rate and locoreglonal control as study endpoints. Materials and Methods : Twenty-two patients with head and neck cancer underwent a FDG-PET study before RT, after a flrst dose of 45 Gy, and after a second dose on more 4han 70 Gy. Standard uptake value (SUV) was calculated for primary tumor (n=22) and neck lymph node (n:10). Attenuation corrected PET scans acquired 60 min after tracer injection were used for evaluation of FDG uptake In tumors. A quantitative FDG uptake index was expressed as Suvlean (corrected for iean body mass). The follow-up time was at least 5 months (range S-1 S months). Results : A total of 22 primary tumors and 10 metastatic lymph nodes were analyzed In FDG-PET. In the first PET study the mean SUVlean the primary tumors and nodes were 5.4 (SD, 2.5) and 4.6 (SD, 2.3), respectively. In the second PET, study peformed after 46 Gy RT the mean SUV in primary tumor and node decreased to 2.9 (SD, 1.9, p<0.001) and 1.7 (SD, 1.3) respectively. in the third PET study peformed at the full dose (more than 70 Gy), RT the mean SUV In the primary tumors and nodes decreased to 2.3 (SD, 1.5, p<0.001) and 1.5 (SD, 1 .1) respectively. Conclusions: FDG uptake In tumors showed a significant decrease after the 45 Gy and more than 70 Gy of RT for squamous cell carcinoma of the head and neck. Reduction of metabolic activity after 46 Gy of radiotherapy Is closely correlated with radiation response.
The purpose of this study was to survey the knowledge and awareness of dental hygienists and dental hygiene students about hepatitis B in an attempt to lay the solid foundation for the prevention of hepatitis B and infection control, as dental hygienists were highly likely to be exposed to HBV during job performance. The findings of the study were as follows: 1. 32.5% of the dental hygienists and 15.6% of the dental hygiene students were aware of their own HBsAg and HBsAb states. The dental hygienists who were cognizant of the states statistically significantly outnumbered the dental hygiene students who were (p=.000). 2. As for vaccination against hepatitis B, 71.8% of the dental hygienists and 47.3 % of the dental hygiene students were inoculated against it. The dental hygienists who were vaccinated against it outnumbered the dental hygiene students who were, and the disparity between the two was statistically significant (p=.000). 3. In the event of those who were vaccinated against hepatitis B, the dental hygienists were better cognizant than the dental hygiene students of the right time for the vaccination, required vaccination frequency (three times) and confirmation of the formation of hepatitis B antibody (p=.000). 4. Regarding awareness of hepatitis B infection route, the dental hygienists knew significantly better than the dental hygiene students that hepatitis B might be infected via blood (p=.030), sexual relations (p=.000), contaminated needle sticks (p=.000), mothers with hepatitis B positive during delivery (p=.000), toothbrushes/razors (p=.000) and exchange of drinking cups (p=.000). 5. As to the relationship between health status and knowledge on hepatitis B infection route, the respondents who were in bad shape had the best knowledge about that, followed by those in an average state of health and healthy respondents. And the dental hygienists had a significantly better knowledge than the dental hygiene students (p=.001). 6. Just a small number of the dental hygienists and students knew about the hepatitis B-related past experiences of their families, and the gap between the two was insignificant.
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