• Title/Summary/Keyword: in vitro detection

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Effects of First Assisted Reproductive Technologies on Anxiety and Depression among Infertile Women: A Systematic Review and Meta-Analysis (1차 체외수정시술이 난임여성의 불안과 우울에 미치는 효과: 체계적 문헌고찰과 메타분석)

  • Ha, Ju-Young;Ban, Seon-Hwa;Lee, Hae-Jung;Lee, Misoon
    • Journal of Korean Academy of Nursing
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    • v.50 no.3
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    • pp.369-384
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    • 2020
  • Purpose: The purpose of this study was to analyze anxiety and depression among infertile women at different time points during the first In Vitro Fertilization (IVF) or Intracytoplasmic Sperm Injection (ICSI) treatment through a systematic review and meta-analysis. Methods: Seven out of 3,011 studies were included for meta-analysis. To estimate the effect size, a meta-analysis of the studies was performed using the RevMan 5.3 program. We compared the measurement outcomes at three time points: before the start of treatment (T0), cancellation of treatment after pregnancy detection (T2), one to six months after treatment (T3). The effect size used was the standardized mean difference (SMD). Results: In comparing the different time points of the pregnant women from their cycle, significantly lower levels of depression were found at T2 than at T0. In non-pregnant women, anxiety at T2 and depression at T2 and T3 were significantly higher than those at T0. At T2 and T3, the non-pregnant women reported higher levels of anxiety and depression compared with the pregnant women. Conclusion: Anxiety and depression in infertile women undergoing the first IVF or ICSI are associated with the time points and pregnancy status after treatment. These findings suggest that attention should be paid to helping infertile women prepare for and cope with treatment and treatment failure.

Production System of Virus-free Apple Plants Using Heat Treatment and Shoot Tip Culture (열처리와 경정배양을 이용한 바이러스 무병 사과 생산 시스템)

  • Lee, Gunsup;Kim, Jeong Hee;Kim, Hyun Ran;Shin, Il Sheob;Cho, Kang Hee;Kim, Se Hee;Shin, Juhee;Kim, Dae Hyun
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.288-293
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    • 2013
  • In worldwide, viral diseases of apple plants has caused the serious problems like reduced production and malformation of fruits. Also, the damages of apple plants by virus and/or viroid infection (Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple mosaic virus, and Apple scar skin viroid) were reported in Korea. However there is few report about the protection approach against the infection by apple viruses. Therefore, this paper introduced the experimental protocol for the development of virus-free apple cultivars (Danhong, Hongan, Saenara, Summerdream). Apple plants were treated at $37^{\circ}C$ for 4 weeks and shoot tips were cultured in vitro. After heat treatment, the detection of apple viruses was performed by RT-PCR using virusspecific detection primers in new apple cultivars. With the heat treatments followed by in vitro shoot tip culture, the proportion of virus-free stocks of 'Danhong', 'Hongan', 'Saenara', and 'Summerdream' was 28%, 16%, 12%, and 12%, respectively. Taken together, this approach can be a good tool for production of virus-free apple stocks.

Optimal Conditions of Single Cell Gel Electrophoresis (Comet) Assay to detect DNA single strand breaks in Mouse Lymphoma L5178Y cells

  • Ryu, Jae-Chun;Kwon, Oh-Seung;Kim, Hyung-Tae
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.89-94
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    • 2001
  • Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

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The Significance of Gonadotropin Ratio in In Vitro Fertilization of Human Oocytes (성선 자극호르몬의 비율이 인간난자의 체외수정에 미치는 영향에 관한 연구)

  • Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.15 no.2
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    • pp.135-147
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    • 1988
  • To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.

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Regulation of Phthalates and Their Alternatives in Children's Products and Their Toxicity Data (어린이제품 내 프탈레이트류 및 대체제의 규제와 독성자료에 대한 연구)

  • Lee, Inhyae;Ra, Jinsung;Ji, Kyunghee
    • Journal of Environmental Health Sciences
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    • v.47 no.1
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    • pp.1-19
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    • 2021
  • Objectives: Phthalates, which are widely used as plasticizers, have been recognized as endocrine disruptors. In the present study, we provided information on the regulation of these chemicals and summarized the information available on their detection and toxicity in children's products and those of their alternatives. Methods: The regulatory frameworks related to phthalates in children's products in Korea, the United States (US), and the European Union (EU) were compared. Data on the detection concentration of 16 phthalates and seven phthalate alternatives that could be used in polyvinyl chloride (PVC) plastic products for children as well as on their toxicity classification and endocrine disruption toxicity were collected from the literature. Results: Korea adopted US and EU chemical standards for six phthalates (DEHP, BBP, DBP, DINP, DIDP, and DNOP), but not others (e.g., DIBP, DPP, DHP, and DCHP). Among the ten phthalates and seven substitutes for which regulatory standards were not determined, DIBP, DHP, DEHA, DIBA, DINA, and DEHT were detected in children's products made from PVC plastic. DIBP and DHP, which have a reproductive toxicity classification of 1B, were frequently detected in PVC toys. The reproductive toxicity, estrogenicity, and anti-androgenic activity of the unregulated phthalates and their alternatives have been reported in diverse in vitro and in vivo assays. Conclusion: The use of unregulated phthalates and their substitutes in children's products is increasing. Further monitoring and toxicological information on phthalate alternatives is required to develop proper management plans.

Application of Cumulus Cells as Factors to Predict the Outcome of IVF-ET (체외수정시술의 결과를 예측할 수 있는 인자로서 난구세포의 활용에 관한 연구)

  • Kim, Kwang-Dae;Kim, Ki-Hyung;Na, Yong-Jin;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.419-432
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    • 1999
  • Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.

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Effect of object position in the field of view and application of a metal artifact reduction algorithm on the detection of vertical root fractures on cone-beam computed tomography scans: An in vitro study

  • Nikbin, Ava;Kajan, Zahra Dalili;Taramsari, Mehran;Khosravifard, Negar
    • Imaging Science in Dentistry
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    • v.48 no.4
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    • pp.245-254
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    • 2018
  • Purpose: To assess the effects of object position in the field of view (FOV) and application of a metal artifact reduction (MAR) algorithm on the diagnostic accuracy of cone-beam computed tomography (CBCT) for the detection of vertical root fractures(VRFs). Materials and Methods: Sixty human single-canal premolars received root canal treatment. VRFs were induced in 30 endodontically treated teeth. The teeth were then divided into 4 groups, with 2 groups receiving metal posts and the remaining 2 only having an empty post space. The roots from different groups were mounted in a phantom made of cow rib bone, and CBCT scans were obtained for the 4 different groups. Three observers evaluated the images independently. Results: The highest frequency of correct diagnoses of VRFs was obtained with the object positioned centrally in the FOV, using the MAR algorithm. Peripheral positioning of the object without the MAR algorithm yielded the highest sensitivity for the first observer (66.7%). For the second and third observers, a central position improved sensitivity, with or without the MAR algorithm. In the presence of metal posts, central positioning of the object in the FOV significantly increased the diagnostic sensitivity and accuracy compared to peripheral positioning. Conclusion: Diagnostic accuracy was higher with central positioning than with peripheral positioning, irrespective of whether the MAR algorithm was applied. However, the effect of the MAR algorithm was more significant with central positioning than with peripheral positioning of the object in the FOV. The clinical experience and expertise of the observers may serve as a confounder in this respect.

Biochemical Reactions on a Microfluidic Chip Based on a Precise Fluidic Handling Method at the Nanoliter Scale

  • Lee, Chang-Soo;Lee, Sang-Ho;Kim, Yun-Gon;Choi, Chang-Hyoung;Kim, Yong-Kweon;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.146-153
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    • 2006
  • A passive microfluidic delivery system using hydrophobic valving and pneumatic control was devised for microfluidic handling on a chip. The microfluidic metering, cutting, transport, and merging of two liquids on the chip were correctly performed. The error range of the accuracy of microfluid metering was below 4% on a 20 nL scale, which showed that microfluid was easily manipulated with the desired volume on a chip. For a study of the feasibility of biochemical reactions on the chip, a single enzymatic reaction, such as ${\beta}-galactosidase$ reaction, was performed. The detection limit of the substrate, i.e. fluorescein $di-{\beta}-galactopyranoside$ (FDG) of the ${\beta}-galactosidase$ (6.7 fM), was about 76 pM. Additionally, multiple biochemical reactions such as in vitro protein synthesis of enhanced green fluorescence protein (EGFP) were successfully demonstrated at the nanoliter scale, which suggests that our microfluidic chip can be applied not only to miniaturization of various biochemical reactions, but also to development of the microfluidic biochemical reaction system requiring a precise nano-scale control.

In vitro Dissolution and in vivo Bioequivalence Study of Controlled Release Carbamazepine Formulation (Epileptol CR® vs Tegretol CR® in Healthy Male Korean Volunteers

  • Kim, Ji-Young;Kim, Hyung-Tae;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.38 no.5
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    • pp.335-342
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    • 2008
  • The bioequivalence of two carbamazepine preparations was conducted. The in vivo bioequivalence study in 20 healthy male Korean volunteers was designed by using a single dose, randomized, 2-period crossover with a 3-weeks washout period between the doses. Prior to the in vivo study, an in vitro comparative dissolution test was performed by the paddle and basket method as described in the bioequivalence guidance of the Korea Food and Drug Administration (KFDA). Based on the similar dissolution pattern between two preparations in the dissolution test, the two formulations are demonstrated to be pharmaceutically equivalent. In addition, in vivo bioequivalence test was used to reconfirm the in vitro dissolution results. In the in vivo bioequivalence study, the plasma concentrations of carbamazepine up to 144 h after the administration were determined using a validated HPLC method with UV detection and the bioequivalence between the two drug products was assessed by statistical analysis of the log transformed mean ratios of $C_{max}$, $AUC_{0-t}$ and $AUC_{0-\infty}$. The mean maximum concentration ($C_{max}$) of the test and reference were found to be $1467.0{\pm}335.8\;ng/mL$ and $1465.9{\pm}310.3\;ng/mL$, respectively. The 90% confidence intervals (C.I.) of $C_{max}$ were in the range from 0.95 to 1.05. As for the $AUC_{0-t}$ and $AUC_{0-\infty}$, test values were $110027.1{\pm}27786.4\;ng/mL{\cdpt}h$, $128807.0{\pm}34563.2\;ng/mL{\cdot}h$ and $105473.6{\pm}26496.2\;ng/mL{\cdot}h$, $125448.5{\pm}35975.5\;ng/mL{\cdot}h$, respectively. The 90% C.I. of $AUC_{0-t}$ were 0.97 to 1.10 and of $AUC_{0-\infty}$, 0.99 to 1.09 and thus were within the log 0.8-log 1.25 interval proposed by the KFDA. A two-way ANOVA showed no significant difference between the two formulations. Based on these statistical analysis, it was concluded that the test formulation is bioequivalent to the reference.

Detection of transgene in early developmental stage by GFP monitoring enhances the efficiency of genetic transformation of pepper

  • Jung, Min;Shin, Sun-Hee;Park, Jeong-Mi;Lee, Sung-Nam;Lee, Mi-Yeon;Ryu, Ki-Hyun;Paek, Kee-Yoeup;Harn, Chee-Hark
    • Plant Biotechnology Reports
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    • v.5 no.2
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    • pp.157-167
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    • 2011
  • In order to establish a reliable and highly efficient method for genetic transformation of pepper, a monitoring system featuring GFP (green fluorescent protein) as a report marker was applied to Agrobacteriummediated transformation. A callus-induced transformation (CIT) system was used to transform the GFP gene. GFP expression was observed in all tissues of $T_0$, $T_1$ and $T_2$ peppers, constituting the first instance in which the whole pepper plant has exhibited GFP fluorescence. A total of 38 T0 peppers were obtained from 4,200 explants. The transformation rate ranged from 0.47 to 1.83% depending on the genotype, which was higher than that obtained by CIT without the GFP monitoring system. This technique could enhance selection power by monitoring GFP expression at the early stage of callus in vitro. The detection of GFP expression in the callus led to successful identification of the shoot that contained the transgene. Thus, this technique saved lots of time and money for conducting the genetic transformation process of pepper. In addition, a co-transformation technique was applied to the target transgene, CaCS (encoding capsaicinoid synthetase of Capsicum) along with GFP. Paprika varieties were transformed by the CaCS::GFP construct, and GFP expression in callus tissues of paprika was monitored to select the right transformant.