Many pronuclear stage eggs were used to generate transgenic mice (Tg) by microinjection. In this study, we used in vitro fertilized mouse eggs, followed by ultrarapid freezing to establish a simple procedure for production of Tg mice. We produced in vitro fertilized mouse eggs and cryopreserved them by ultrarapid freezing method. A total of 139 cryopreserved-thawed pronuclear eggs, of which 101 (72.6%) were survived following microinjection of chicken b-actin promoter-driven firefly improved luciferase cDNA (β-act/luc+) and were transferred into 5 recipients. All recipients became pregnant and gave birth to a total of 15 (14.8%) pups. As a control, same DNA construction (β- act/luc+) was also injected into 450 in vitro fertilized eggs, of which 338 (75.1%) were survived and then were transferred into 14 recipients. Eleven (78%) mice became pregnant and littered a total of 54 (19.1%) pups. Southern blotting analysis of Tg mice indicated that one (1/15, 6.6%) and three (3/54, 5.5%) transgenic mice were production from cryopreserved and in vitro fertilized eggs, respectively. All Tg mice produced from both eggs showed the expression of improved luciferase gene. These results indicated that efficiency of produced of Tg mice from cryopreserved eggs was comparable to that from in vitro fertilized eggs. Furthermore, it is suggested that microinjection of transgene into in vitro fertilized eggs cryopreserved by ultrarapid freezing is an easy and conveniently method for production of Tg mice.

Selenium (Se) is an essential trace element for humans and animals, and several findings suggest that dietary Se intake may be necessary for bone health. Accumulating evidence indicates that Se compounds possess anticancer properties. Se is specifically incorporated into proteins in the form of selenocysteine and non-specifically incorporated as selenomethionine in place of methionine. This study evaluated protection by Se in the bone repair process in ovariectomized rats after irradiation. For such purpose, 80 ovariectomized female Sprague-Dawley rats were randomly divided into 4 experimental groups: ovariectomized (Ov), Ov/Se, Ov/irradiated (Irr) and Ov/ Se/Irr. A bone defect was created on the tibia of all animals 40 days after ovariectomy. Two days after surgery, only the Ov/Se and Ov/Se/Irr rats received 0.8 mg Se/kg. Three days after surgery, only the Ov/Irr and Ov/Se/Irr rats received 10 Gy of X-rays on the lower limb region. The animals were euthanized at 7, 15, 22 and 29 days after surgery to assess the repair process, which was evaluated by analysis of trabecular bone number (Masson Trichrome) and birefringence analysis (Picrosirius). It was possible to observe a delay in the bone repair process in the ovariectomized/irradiated group and similarity between the ovariectomized, Ov/ Se and Ov/Se/Irr groups. Our findings suggest that sodium selenite may influence a radioprotective effect in the bone repair of tibia of ovariectomized rats without toxicity.

The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG (81.1±15.1), 1,2-PROH (88.0±21.1) or the control (99.9±21.3) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO (14.7±1.3), EG (12.1±1.1), Glycerol (15.2±1.8), and 1,2-PROH (11.5±1.3) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.5±0.7, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

The objective of this study was to monitor health conditions of genetically identical somatic cells cloned Korean white cattle, endangered indigenous cattle (EIC) and indigenous cattle (IC) by analysis of hematologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and one live cattle were delivered by caesarean section. The cloned Korean white cattle were genetically identical to the nuclear donor cattle. As a result, the mean values of RBC and platelet of cloned cattle and white cattle were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between cloned cattle and IC of the same age (1∼2 years) showed the statistical significance (P<0.05). Also, in the WBC of Korean white cattle, the estimated values were decreased according to the age from 12.0×103/μl under 1 year to 11.0×103/μl over 1 years respectively. Although clone-cattle had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned cattle were not different when compared to reference range. This study suggests that cloned Korean white cattle derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological parameters. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

이 연구에서는 정소가 제거된 흰쥐에 비타민 E와 셀레늄(Selevit)을 5주간 투여 후 체중, 장기 무게, 혈액학적 그리고 생화학적 변화를 관찰하였다. 체중의 변화에서는 모든 실험군에서 증가가 나타냈다. Orch+Selevit군의 체중 증가는 11.2±10.25 g으로 가장 낮았으며, Intact군, Sham군, Orch군과 비교 시 유의적으로 감소되었다. 장기 무게의 변화에서는 Orch+Selevit군의 심장과 간장 무게는 Intact군, Sham군과 비교 시 유의적으로 감소되었다. Intact군, Sham군, Orch군의 신장 무게는 Orch+Selevit군 비교 시 유의적인 증가를 확인하였다. 백혈구 수의 혈액학적 변화에서는 Orch+Selevit군은 다른 모든 군과 비교해 유의적인 증가를 확인하였으며, 적혈구수, 평균적혈구용적, 평균적혈구혈색소량, 평균적혈구색소농도 등과 같은 혈액학적 측정치에서는 유의성은 인정되지 않았다. 생화학적 변화에서는 Orch+Selevit군의 혈청총단백질, 알부민은 Orch군과 비교 시 유의적으로 증가하였으며, 알부민은 Intact군, Sham군 그리고 Orch군과 비교 시 유의적으로 감소했다. AST와 ALT는 모든 실험군에서 유의성이 없었다.

본 연구에서 배아의 생식세포 동결에 가장 흔히 쓰이고 있는 두 가지 동결 보호제, 즉 DMSO와 EG의 독성을 비교하고자 생쥐 수정란 모델을 이용한 실험을 하였다. 생후 6주령의 암컷 생쥐 F1 hybrid mice에 10 IU의 PMSG를 복강 주사하여 과배란을 유도하고, 2-세포기 배아를 획득하고 DMSO와 EG 각각 노출시킨 후, 배양을 하였다. 배반포의 전체 세포수는 2-세포기 단계에서 DMSO(68.1±24.1)로 EG(81.2±27.0) 혹은 control(99.0±18.3)(p<0.001) 처리구에 비해서 유의적으로 낮았다. DMSO 처리구가 EG 처리구에 비해 세포수가 적었다. DMSO(15.4±1.5)와 EG(10.2±1.4) 두 처리구는 대조구(6.1±0.9, p<0.0001)와 비교해서 배반포에서 세포사 비율이 더 높음을 확인했다. 또한, DMSO 처리구는 EG 처리구(p<0.001)보다 더 많은 세포사멸된 세포가 확인되었다. DMSO 또는 EG 처리군과 대조군 사이에는 배아 부화율에 있어서 차이가 있었으며, 이는 배아에 대한 동결 보호제의 잠재적인 독성을 확인한 결과였다. 이번 연구에서 장기간 처리했을 때 EG 처리군보다 DMSO 처리군에서 배아발달과 세포수가 저하된 것은 DMSO의 독성이 더 높을 것으로 사료된다.

Despite the increasing interest in noticing by second language (L2) learners during the output process, little attention has been given to child L2 learners’ noticing and output. Thus, the present study investigated what child L2 learners of different English proficiency levels noticed as they composed a text and received written corrective feedback (WCF) (error correction vs. models), and how they incorporated the noticed features into their revised texts. Data were collected from twenty-six child pairs throughout three stages (composition, comparison, and revision stages) and note-taking was employed as a means of measuring learner noticing. It was found that learners were able to initiate noticing on their own when composing texts, and that highproficiency learners tended to attend to grammatical problems more frequently than medium/low-proficiency learners. WCF played a facilitative role in leading learners to notice, and their attention to language was mediated differently by different types of WCF. Learners incorporated the noticed features into their revisions, and textual revisions were mostly lexical. Error correction (EC) triggered more grammatical revisions, while the model text helped learners to notice something beyond what they could produce on their own. These findings suggest a facilitative role for output and WCF as a means of learner noticing, and the different roles of EC and model texts.

담관 협착의 원인을 양성인지 악성인지 구별하기가 어려운 경우가 종종 있어 많은 임상의사들을 곤혹스럽게 한다. 그러한 이유는 여러 가지 영상검사와 내시경적 역행성 담췌관 조영술에 의한 반복적인 세포검사와 조직검사를 시행함에도 불구하고 양성 담관 협착과 악성 담관 협착을 수술 전 정확히 감별하지 못하는 경우가 많기 때문이다. 저자들은 50 세 남자환자에게 원위부 담도 폐쇄 소견이 있어 세포 검사와 조직검사를 여러 차례 반복적으로 시행하였으나 악성세포는 나오지 않았고, 간흡충 충란이 다량 발견되어 간흡충증에 의한 양성 협착과 악성 협착의 감별이 어려운 상황에서 1개월 후 복부 전산화 단층촬영을 시행하였다. 그 결과 췌장실질의 위축과 췌관확장이 진행하여 악성종양을 강력히 의심하고 유문부 보존 췌십이지장 절제술을 시행하였다. 조직검사 결과에서 췌장 머리의 선암으로 진단하였다. 악성 담관 협착을 수술 전에 정확히 진단하기는 쉽지 않지만, 담관 협착은 악성이 의심되면 수술적 진단 등의 적극적 검사를 주저해서는 안될 것이다.

림프구는 면역기능의 중심적인 역할을 담당하는 세포이고, 운동에 의해 변화하고 노화에 의해 기능이 저하하는 것으로 잘 알려져 있지만, 고령자를 대상으로 운동에 의한 림프구의 변동에 대한 연구는 지금까지 미흡한 실정이다. 그래서 본 연구에서는 운동훈련이 고령흰쥐의 림프구에 미치는 영향에 대해 조사하였다. 8주간의 운동처리가 비장세포 중 그리고 장간막 림프절의 림프구에 미치는 영향을 분석했다. 비장세포중의 림프구에 대해서는 헬프-T 림프구, 세포상해성 T 림프구, 그리고 보조자극 신호 수용체 발현 T 림프구의 비율이 운동처리구에 있어 유의적으로 높은 수치를 나타냈다. 한편, 비장세포중의 B 림프구 및 장간막 림프절중의 림프구에 대해서는 두 처리구 간에는 유의적인 차이는 보이지 않았다. 이상의 결과들로부터, 운동처리구가 흰쥐 비장세포중의 T 림프구수에 영향을 미치는 것이 밝혀졌다. 그러나 면역기능을 평가하기 위해서는 림프구의 수 뿐만 아니라 기능에 대해서도 면밀히 검토해야 한다. 금후에는 운동처리구가 림프구의 기능에 미치는 영향을 해석할 필요성이 사료된다.

Task-based language instruction has been increasingly promoted in second language (L2) classrooms under the assumption that it can provide an ultimate condition for L2 learning when combined with form-focused interventions such as teacher corrective feedback (CF). The present research investigated the amount, type, and learner uptake of teacher CF in relation to the following task-related variables: (1) task vs. nontask work, (2) focused vs. unfocused tasks, and (3) open vs. closed tasks. Data were collected from 8 intermediate adult EFL classrooms. It was found that the amount of teacher CF provided during tasks versus nontasks and focused versus unfocused tasks was not significantly different. However, the teachers used different strategies depending on the focus of tasks. The study did not relate the matter of open versus closed tasks to the amount and types of teacher CF. When considering learner uptake, the study found that the teachers tended to offer more uptake opportunities during relatively form-oriented activities, and the learners’ uptake was more successful when they were working in relatively form-oriented activities compared to the cases where they were in meaning-based activities. Learner uptake was not associated with whether or not tasks were open and closed.

The importance of genetic resource preservation has been highlighted in the literature as a means of maintaining genetic diversity. Investigations for hematologic values and the differential count of white blood cell count (WBC) for Korean indigenous cattle (KIC) and endangered indigenous cattle (EIC) are rarely performed. Therefore, the objective of this study was to investigate the hematologic values of total 40 EIC (White, Black, Mini cattle) and 35 KIC as control by analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of EIC were significantly decreased by age (p<0.05). The mean values of RBC, HCT, MCV and MCHC between EIC and KIC of the same age (2 ～3 years) showed the statistical significance (p<0.05). Also, in the WBC of EIC, the mean values were decreased according to the age from 13.9×103/μL～12.7×103/μL under 1 year to 9.1×103/μL～11.5×103/μL over 2 years respectively. In the differential count of WBC of EIC (White, Black, Mini cattle), it showed generally the rates of 40.2%, 52.2%, 49.0% lymphocyte and 27.2%, 33.9%, 32.0% segmented neutrophil from 2～3 years respectively. Result of this study will be used for establishing reference range for blood analysis in EIC such as white, black and mini cattle. This study reported hematological values which could serve as baseline information for comparison in conditions of nutrient deficiency, physiological and health status of endangered Korean native cattle. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

Articles in one domestic journal, English Teaching (ET), and one international journal, TESOL Quarterly (TQ) from 2011 to 2013 were surveyed in order to analyze current research trends and to suggest better orientation toward future research focusing on participants, research methods, target language skills, and research topics. The results show that university students have participated most in both of the journal articles, indicating that they are practically available to the researchers in higher educational settings. The proportion of quantitative and qualitative research methods was well balanced in TQ, while, in ET, more than 50 percent of the articles relied on quantitative research methodology along with a relatively larger proportion of mixed method research in ET. In terms of target language skills, writing has been studied most in ET and speaking in TQ, showing growing interest in productive language skills nowadays. As for research topics, articles based on socio-cultural factors have been published most in TQ and articles based on classrooms pedagogy in ET, which reflects that English learning is performed mostly in classroom settings in Korea. Pedagogical implications and suggestions are made based on the findings

This study was conducted in order to examine relationship between stroke patients' degree of satisfaction with leisure and quality of life according to their leisure activity types and whether they conducted leisure activities. A direct survey was performed from April 8 to May 3, 2013 on 92 inpatients and outpatients who received occupational therapy at hospitals located in Jeonju and Gunsan. A frequency analysis was carried out in order to look at the general characteristics of the subjects, their degree of participation in leisure activities, and their degree of satisfaction with leisure activities. A Pearson's coefficient was used to examine relationship between their degree of satisfaction and quality of life according to participation in leisure activities. Their degree of satisfaction according to participation in leisure activities was significantly higher when they took part in leisure activities such as handicraft activities, sports activities, or travel or tourism activities than when they did not perform any leisure activities. Their quality of life according to participation in leisure activities was higher when they conducted sports activities, outing activities, or tourism activities than when they did not. There was significant correlation between their degree of satisfaction with leisure and quality of life according to leisure activities.

The goal of this study was to assess the effect of a group exercise program on cognitive function of elderly people. Subjects were chosen to be elders with dementia having minor to moderate degrees of cognitive function. Study was started out by randomly dividing the 16 subjects into two groups, each with 8 people; the group exercise group and the control group. The group exercise group performed 8 weeks of group exercise program and general physiotherapy while the control group only performed general physiotherapy. Cognitive function was measured by Korean version mini-mental state examination. The study group's attention and calculation statistically significantly improved but the control group saw no statistically significant change. The group exercise program affected improvement in cognitive function of elderly people with dementia and in particular, was effective for enhancing their attention and calculation.

The insecticidal activities of materialsderived from Dryopteris crassirhizoma Nakai against third instar larvae of three species mosquitoes (Culex pipiens, Aedes albopictus and Anopheles sinnensis) were evaluated using a direct contact mortality bioassay. The methanol extracts of D. crassirhizoma showed 100%, 87.8% and 100% larvicidal activity at 1,000 ppm against Cx. pipiens, Ae. albopictus and An. sinensis, respectively. Hexane fraction showed 100% larvicidal activity three species mosquitoes at 500 ppm after 24 hrs. Purification of the biologically active constituents from the hexane extraction with larvicidal activity was done using silica gel column chromatography. H1 and H3 fractions gave 100% mortality to Cx. pipiens, Ae. albopictus and An. sinensis at 100 ppm. H1 fraction separated with methanol to give a H111 through centrifugation. Fraction of the biologically active constituents from the H3 fraction with larvicidal activity was done using methanol. H31 fraction was determined 100% mortality to Cx. Pipiens, Ae. albopictus and An. sinensis at 50 ppm, respectively. Two fractions were analyzed C14H22O (H111, MW206.0) and C11H14O4 (H31, MW210.08) by GC and GC-MS. D. crassirhizoma derived compounds described herein could be useful for managing filed populations as larvicide of Cx. pipiens, Ae. albopictus and An. sinensis.

This study was conducted for SNPs in the 5'-regions of estrogen receptor-α(ESR-α), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genet-ic improvement was incurred low reproduction performance. The objective of this study was to investigate connec-tions between single nucleotide polymorphisms (SNP) of Estrogen receptor-α (ESR-α) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to ge-notype the cattle. The result showed two variants of the ESR-α gene. The Bgl I cut the 492 bp amplification pro- duct into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction frag-ments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in the-se breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was 382.18±10.03 days, and G/G was 381.69±5.22 days. The SPC of A/G was 1.62±0.16, and G/G was 1.32±0.04. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was 7,253.00±936.00 kg and of G/G was 8,747.51±204.88 kg, showing no significant difference.

The legume family (Fabaceae) is the third largest family of flowering plants worldwide and often damaged by stink bugs, particularly Riptortus clavatus Thunberg (Heteroptera: Alydidae). This insect selectively feeds and oviposits on these plants, resulting in >50% of damage to soybeans harvested in autumn. The most common control methods for this insect are periodic application of chemical pesticides, pheromone traps, and bait crops, but still they encounters residual problem, insect resistance or low control efficacy. The current management system needs alternative control methods, which possibly combined with the present methods. Some entomopathogenic fungi have high virulence against the nymphs and adults of R. clavatus in a favorable conditions, reaching ∼80% mortality with mycosis in 3∼5 days. The genera of Beauveria and Metarhizium are possible control agents for effective and safe management of the serious pest. Fungal infection in R. clavatus was observed using an egfp-expressing fungal transformant. Thermotolerant entomopathogenic fungal conidia can be produced in cereal substrates, which enables conidia to be stable for long times. Our considerations need to be given to the combinations with other control methods, such as chemical pesticides or pheromone trap.

The objective of this study was to examine the effects of auditory biofeedback training and kicking training on walking times in patients with hemiplegia to determine if the effects of such training would be maintained even after stopping the intervention. Thirty hemiplegia patients were selected and randomly assigned to a control group of 10 patients to receive general exercise treatment; an auditory biofeedback training group of 10 patients to receive auditory biofeedback training, along with general exercise; and a kicking training group to receive kicking training, along with general exercise. All the patients received their respective training 12 times in total, three times per week for four weeks. In addition, all those in the control and experimental groups received the same general exercise treatment 12 times in total, three times per week for four weeks, and underwent follow-up tests thereafter. The patients’10m walking times were measured using a stopwatch. The significance was analyzed using repeated-ANOVAs. In cases where there were interactions between measuring times and groups, in each group were examined using repeated-ANOVAs. In cases where there were differences, post-hoc tests were conducted using repeated of contrast test. The 10m walking times of the control and experimental groups were significant differences in 10m walking times were shown between measurement times(p<.05), and significant differences in the interactions between measuring times and groups were shown between the groups(p <.05). However, no significant differences in 10m walking times were shown between the groups(p>.05). The auditory biofeedback training group showed significant decreases in walking times four weeks after the beginning of the intervention(p<.05) and significant increases eight weeks after the beginning of the intervention(p<.05). The kicking training group showed significant decreases in walking time four weeks after the beginning of the intervention(p<.05) and maintained the walking times without showing any significant differences eight weeks after the beginning of the intervention(p>.05). The walking speeds of only the kicking training group were maintained until eight weeks after the beginning of the intervention.

The objective of this study was to examine the effects of auditory biofeedback training and kicking training on walking times in patients with hemiplegia to determine if the effects of such training would be maintained even after stopping the intervention. Thirty hemiplegia patients were selected and randomly assigned to a control group of 10 patients to receive general exercise treatment; an auditory biofeedback training group of 10 patients to receive auditory biofeedback training, along with general exercise; and a kicking training group to receive kicking training, along with general exercise. All the patients received their respective training 12 times in total, three times per week for four weeks. In addition, all those in the control and experimental groups received the same general exercise treatment 12 times in total, three times per week for four weeks, and underwent follow-up tests thereafter. The patients’10m walking times were measured using a stopwatch. The significance was analyzed using repeated-ANOVAs. In cases where there were interactions between measuring times and groups, in each group were examined using repeated-ANOVAs. In cases where there were differences, post-hoc tests were conducted using repeated of contrast test. The 10m walking times of the control and experimental groups were significant differences in 10m walking times were shown between measurement times(p<.05), and significant differences in the interactions between measuring times and groups were shown between the groups(p <.05). However, no significant differences in 10m walking times were shown between the groups(p>.05). The auditory biofeedback training group showed significant decreases in walking times four weeks after the beginning of the intervention(p<.05) and significant increases eight weeks after the beginning of the intervention(p<.05). The kicking training group showed significant decreases in walking time four weeks after the beginning of the intervention(p<.05) and maintained the walking times without showing any significant differences eight weeks after the beginning of the intervention(p>.05). The walking speeds of only the kicking training group were maintained until eight weeks after the beginning of the intervention.

The purpose of this study is to provide the case of influence of myofacial release(MFR) and cryotherapy in acute whiplash injury. The subjects were consisted of 3 adults(2 males, 1 female) who had been diagnosed with whiplash injury. The performance period was 1 weeks, 6 times and 1 time is 40min(MFR: 20min, cryotherapy: 20min). We measured neck pain(Visual analogue scale, VAS) and cervical alignment(cervical curvature & line of gravity). All measurements of each subject were measured at pre-treatment and post-treatment. The results are listed below. The neck pain(VAS) index decreased from 7.54cm to 4.11cm. The cervical curvature increased from 23.62°to 29.76°. The line of gravity decreased from 8.03mm to 3.66mm. Based on the results, it can be suggested that both MFR and cryotherapy can be used to neck pain and alignment in acute whiplash injury.