For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at — 7℃ to — 35℃ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen- thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/ 0.5 ml, and 43.6 % frozen at 20embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

Superovulation, or ovarian stimulation is a commonly used ART for treatment of human infertility/subfertility. Recent studies suggest that superovulation unaffects methylated imprints acquisition in mouse oocytes during oogenesis, whereas disrupts DNA methylation maintenance in embryos during preimplantation development. However, the mechanisms of defects in methylation maintanence caused by superovulation remain largely unclear. We hypothesized that superovulation may disrupt the expression of DNA methyltransferases (Dnmts), the enzymes which catalyze DNA methylation acquisition and maintenance. The mice were subjected to superovulate with low (6 IU) and high (10 IU) dosage hormone. We examined the global DNA methylation levels in zygotes and DNA methylation of repeated sequences (IAP and Line 1) in blastocyst stage embryos. In addition, we investigated the expression of Dnmts (Dnmt3a, Dnmt3b, Dnmt3l and Dnmt1o) in ovulated oocytes and zygotes. Through staining with antibody 5mC and Di-H3K9 coupled with confocal microscopy, we found that global methylation profiles in zygotes derived from females after low or high dosage hormone treatment were not affected when compared to control counterpart. Moreover, methylation at IAP in blastocysts also was unaffected by superovulation, irrespective of hormone dosage. In contrast, methylation level at Line 1 decreased when the females were administered by high dosage hormone. Furthermore, expression of de novo DNA methyltransferase Dnmt3a, Dnmt3b, Dnmt3L, as well as maintenance Dnmt1o in MII oocytes and zygotes was not disrupted by superovulation. Given superovulation adversely affected methylation maintenance in blastocysts during preimplantation development but with normal expression of Dnmts in oocytes and zygotes, it is indicated that defects of embryonic methylation didn’t originate from abnormal expression of Dnmts.

Cryopreservation of avian semen is a useful tool to preserve genetic resource for aim of preventing extinction induced by infectious disease like avian influenza. Unlike those of mammals, data from chicken cryopreserved semen has not been showed feasible results. So, various cryoprotectants and diluents have been examined in many methods. In this report, as a major ingredient of avian seminal plasm, glutamine was substituted by alanyl glutamine to enhance physiological stability of chicken semen during freezing. We studied effect of glycerol and Dimethylacetamide(DMA) on motility and progressive motility of spermatozoa using glutamine diluent(EK-G) or alanyl glutamine diluent(EK-A) condition. The semen of Ogye was collected twice a week by the dorso-abdominimal massage method and diluted with same volume of EK-G or EK-A at 25℃ and stored for 10 min at 4℃ in cold chamber. Glycerol or DMA was added to diluted semen to reached 7% of final concentration at 4℃. After 3min of equilibration, the diluted semen was packed into 0.25ml straws and subjected to cryopreservation used freezing equipment. The packed straw were placed on height 5 cm above surface of liquid nitrogen(LN2) and held for 10min. After preserved for 2 weeks, the straw was thawed onto the 4℃ cooling bath. The images of motility and progressive motility spermatozoa were recorded by digital image recorder and analyzed by manual. The results showed 68.5% motility and 34.1% progressive motility in DMA/EKA diluent, 31.45% and 17.6% in glycerol/EKA, 45.4% and 8.6% in DMA/EKG, and 9.7% and 6.4% in glycerol/EKG. With these results, the alanyl glutamine and DMA could be used as a main composition of diluent and cryoprotectant for cryopreservation of chicken semen.

The objective of this study was to assess the effect on post-thawed sperm motility, viability and acrosome integrity of boar semen frozen in the freezing extender with chicken or duck egg yolks. The Sperm rich fraction of ejaculates from three Duroc boars were collected by a glove-hand technique. Samples with more than 80% motile sperm were used for this experiment. Semen was diluted with freezing extender (LEY) containing 11% (v/v) lactose, 20% (v/v) hen egg yolk with 3.5% (v/v) glycerol, and 0.5% (v/v) Orvus Es Paste(OEP, Nova Chemical Sales Inc., Scituate, MA. USA) to yield a final sperm concentration of 5×108 cells/ml. Following complete dilution, semen samples were loaded in 0.5 ml French medium straws (IMV technologies, France) and transferred to programmable semen freezer (SY-LAB Gerate GmbH, Austria). For freezing the semen samples, each straw was cooled from 5℃ to — 5℃ at 6℃/min, auto-seeding at — 5℃ and held for 60sec, samples were then cooled from — 5 to — 80℃ at 40℃/min, and thereafter from — 80℃ to — 150℃ at 60℃/min. The yolks used were sourced from fresh chicken and duck eggs. To evaluate the post-thaw sperm quality, semen was thawed at 38℃ for 20 sec and sperm motility, viability and acrosome integrity were assessed. Motility was assessed for %motile cell characteristics using computer-assisted semen analysis (CASA; SAIS SI-100, Medical supply, Korea). The percentage of sperm viability was assessed using LIVE/DEAD® sperm viability kit (Molecular probes, Eugene, OR, USA). The acrosome integrity was assessed by FITC-PNA staining. Sperm quality in terms of motility, viability and acrosome integrity showed higher after freezing in medium containing duck yolk than chicken yolk. However, there was no significant difference in sperm quality for the different types of yolk(p>0.05). * The result of this study showed that there was no significant difference between the egg yolk types when considering the sperm motility, viability and acrosome integrity of boar semen frozen in the freezing extender with chicken or duck egg yolks.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The purpose of this study was to compare the muscle activity of the abdominal and lumbar multifidus during unilateral prone hip extension on the floor and on a round foam roll. Fifteen healthy participants were recruited. They were instructed to perform a unilateral hip extension on the floor and on a round foam roll in the prone position. Surface electromyography (EMG) signals were recorded from bilateral lumbar multifidus (LM), external oblique (EO), and internal oblique (IO) muscles. A paired t-test was used to compare muscle activity, with the level of significance set at =.05. The results showed that bilateral LM, EO, IO EMG activity during right-hip extension on a round foam roll was greater than that on the floor, and EMG activity of bilateral LM, right EO, and left IO during left-hip extension on a round foam roll was greater than that on the floor (p<.05). These findings suggest that the unilateral hip-extension exercise on a round foam roll can be used to activate the lumbar multifidus and abdominal oblique muscles and causes a different increasing pattern between the two lifting sides.

The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

This study aims to examine the effect of patellar taping common to patients with patellofemoral pain syndrome on the change of knee joint location. The total number of participants is 12 patients with no pain in their knee. There are three different experiments: no-taping, placebo taping, and patellar taping. After application, they squat on their hams. As a result, both the muscle activity of vastus medialis and that of vastus lateralis increased in placebo taping compared to no-taping, which wasn't statistically significant. However, the muscle activity of vastus medialis and that of vastus lateralis decreased in patellar taping compared to no-taping, which was statistically significant. This suggests that patellar taping causing the lateral attraction of knee joint is more influential to the dynamics of knee joint than skin afferent input in placebo taping. Therefore, patellar taping is effective to change the location of knee joint, affect the muscle activity of quadriceps muscle of thigh, and thus correct the misalignments of the knee joint.

The purpose of this study was to examine the degree of knowledge of care workers working at long term care hospitals and nursing homes on pressure ulcer. A total of 81 care workers including 34 at long term care hospitals and 47 at nursing homes were surveyed. 24 questions were used to evaluate their degree of knowledge on pressure ulcer. Their knowledge on pressure ulcer scored 12.84 out of the total score of 24 points(SD=3.40), which was equal to 53.50 (SD=24.23) out of 100 points. Their knowledge on the prevention of pressure ulcer was highest among the subareas of evaluation. Their knowledge on pressure ulcer statistically significantly differed according to education on pressure ulcer(P<.05). A pressure ulcer is a skin disorder that may be prevented and cured. At this point when long term care facilities are rapidly increasing, care workers highlevel knowledge on and good management of pressure ulcer is very important. Practically educating them on pressure ulcer including the provision of recent, updated relevant knowledge will be necessary.

This study was carried out to identify how a self-stretching exercise program affects pain for each body area, pain relief and job satisfaction for care workers. 20 of 40 care workers with musculoskeletal symptom were randomly selected and participated a self-stretching exercise program consisting of 15 motions. The intervention was done five times or more per weeks for 8 weeks and 1 session lasted within 15 minutes. 'Musculoskeletal symptom survey table' of the Korea Occupational Safety and Health Agency(KOSHA) and JDI(Job Descriptive Index) was used for pain on the musculoskeletal symptom and job satisfaction. Survey were done twice before and after the program. The result of this study showed that self-stretching exercise program group(SSPG) relieved from pain significantly in the shoulders(p<.01) and lumbar(p<.05), comparing to the non selfstretching exercise program group(NSPG). Although no significant difference on variations in the JDI appeared in SSPG, the significant reduction appeared from the colleague relationship and organization in NSPG(p<.05). SSPG showed the significant increase on variations in JDI from the job and organization comparing to NSPG. Especially, the improvement on satisfaction for the organization was shown(p<.05). Accordingly, the self-stretching exercise program for care workers can be said to positively affect the overall pain relief and increase on the JDI.

For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at —7 ℃ to —35 ℃ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen-thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/ 0.5ml, and 43.6 % frozen at 20 embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

Experiments were conducted in order to assess the healing effect of bee venom (BV) cream on full-thickness skin wounds in rabbits. BV cream was compared with silver sulfadiazine (SS) as a topical medicament against a control on experimentally created full-thickness wounds. Two wounds measuring 2 × 2 cm were created bilaterally (four wounds/rabbit) on the dorsolateral aspect of the trunk of seven New Zealand white rabbits. Wound treatments were evenly distributed on four sites, using a Latin square design. The contact layer of wounds was treated with physiological saline (control), SS cream, and BV cream over a period of 28 days. Assessment of wound healing was based on scab hardness, wound exudates, wound area, unepithelialized granulation tissue, and histopathological findings. Topical application of BV and SS creams to wounds resulted in reduced inflammation, debridement of necrotic tissue, and promoted granulation and epithelialization. Wound healing was faster, with statistical significance in BV and SS treatments, compared to the control (P<0.05). Treatment with BV evoked an anti-inflammation effect in a rabbit model. BV cream produced a wound healing effect similar to that of commercially available SS cream. Anti-inflammation effect as a topical treatment with BV cream appears to be better than that with SS cream. These results suggest that topical application of BV cream may be an alternative treatment for full-thickness skin wounds.

Malignant pleural effusion (MPE) and blood samples can be used as a practical source for detection of epidermal growth factor receptor (EGFR) mutations in patients with advanced non-small cell lung cancer. We compared EGFR mutation status of cell blocks, cell-free fluid of MPE, and plasma from patients with lung adenocarcinoma. We obtained paired samples of MPE and plasma from 14 pathologically-confirmed lung adenocarcinoma patients. Peptide nucleic acid (PNA)-mediated real-time polymerase chain reaction (RT-PCR) clamping was performed for determination of EGFR mutation status. EGFR mutations were detected in five (35.7%) cell blocks of MPE, which showed results identical to those of the corresponding cell-free fluid, whereas mutations were detected in the plasma of only two (40.0%) of the five patients. Of seven patients treated with EGFR tyrosine kinase inhibitors (TKIs), EGFR mutations were detected in cell blocks, cell-free fluid of MPE, and plasma for only one of the four patients who responded to EGFR TKIs, while mutations were detected only in cell blocks of MPE and cell-free fluid of the three remaining patients. Our results suggest that detection of EGFR mutations in cell-free pleural fluid from lung adenocarcinoma patients using highly sensitive methods may be feasible, but that analysis of free plasma may lead to undetected mutations and misdiagnosis.

Cytokines are known to function as regulatory molecules that can be produced by virtually every nucleated cell type in the body, including lymphocytes, monocytes/macrophages, epithelial cells, fibroblasts, and many others. Cytokines include lymphocyte-derived factors (lymphokines), monocyte-derived factors (monokines), hematopoietic factors (colony-stimulating factors), connective tissue/ growth factors, and chemotactic chemokines. Cytokines released in response to infection can affect tumor development in different ways. When exposed to infectious agents, cytokines are secreted by sentinel cells, such as macrophages and dendritic cells. These cytokines include interleukin 1 (IL-1) and tumor necrosis factor-α, as well as others, such as IL-6, IL-12, and IL-18. When released in sufficient quantities, these molecules can cause inflammation. Chronic inflammation is highly associated with tumor initiation, promotion, and progression. In this article, we review the roles and mechanisms of cytokines in tumor development.

음식물 폐기물 침출수를 처리하는 분리막 결합 고온 혐기성소화공정(생물학적 반응조) (Anaeorobic Membrane Bioreactor, AnMBR)의 파일럿 운전에서 분리막의 교차여과 속도와 막간압력이 파울링에 미치는 영향을 관찰하였다. 연구 결과 정압여과 하에서 교차여과 속도가 증가할수록 파울링의 속도는 현격히 감소되었다. 그러나 이와 같은 영향은 낮은 막간압력에서 더욱 효과적이었다. 막간압력이 증가할수록 여과대상 물질의 압축성으로 인해 투과성이 상대적으로 낮은 파울링층(혹은 케익층)이 분리막 표면에 형성된 것에 기인된 듯하다. 여과대상 시료의 입도분석을 해 본 결과 입자크기는 약 10~100mum 범위에서 분포하였고 이에 따라 브라운확산에 의한 역수송보다 분리막 표면에서 교차여과에 의해 발생하는 전단력이 입자의 역수송에 더욱 기여하고 있음을 예측할 수 있었으며 이는 AnMBR의 연속운전을 통해 재확인할 수 있었다. 운전 후 막 부검을 실시한 결과 유기 및 무기 파울링이 모두 관찰되었으나 어느 것이 지배적인 파울링 기작을 나타내는지는 앞으로 더욱 연구가 필요하다. 무기 파울링의 경우 대부분 분리막 표면에서 스케일링 형성이 지배적이었으며, 따라서 분리막의 공극 막힘에 주로 기여하는 작은 콜로이드성 유기물질의 경우 분리막 표면에서 전단력에 의한 역수송 효과는 그다지 크지 않을 것으로 사료된다.

Antimicrobial peptides are widely found in living organisms and are known to play a critical role in innate immunity. Numerous antimicrobial peptides from diverse species appear to be effective against pathogenic microorganisms of bacteria, fungi, protozoa, and viruses. Because antibiotic resistance is a global health issue in the fight against pathogenic microorganisms, there has been an urgent need for development of new antibiotic substances. In the current study, we performed yeast two hybrid screening using Beclin1 bait in order to find new peptide antibiotics from a random peptide library. Two candidate peptides from the screening were expressed in a yeast secretory system of Pichia pastoris and tested for any antimicrobial activity against Staphylococcus aureus, MRSA, MRSA2242, MRSA2250, Lactobacillus casei, and Lactobacillus acidophilus. Disc clear zone assay and spectrophotometric analysis revealed that the two peptides exert a decent activity against the pathogenic bacteria, in contrast to minimal effect on the commensal Lactobacillus strains. Taken together, this study presents novel peptides with antibacterial activity against the pathogenic forms of Staphylococcus aureus and suggests the possibility that these peptides, upon further characterization, may be developed as clinically useful antibiotics.

Beauveria bassiana is one of universal insect pathogenic fungi that have been used for biocontrol agent against insect pests. This fungus has also been studied for medicinal use. To meet for commercial use, the artificial production of the fruit body of this fungus has been established by the Mushtech Co in Korea. This study was carried out to define the morphological features of the fruit body of B. bassiana developed through artificial cultivation. For the observation of mycelia growth, B. bassiana was cultured on the Sabouraud Dextrose agar plus Yeast Extract(SDAY), nut-supplemented medium, and Fe ion-supplemented SDAY at 25℃ for 15 days. The variation of colony color was observed between the different media. Strong pigmentation was observed on Fe ion-supplemented SDAY. To investigate morphological characteristics of fruit body, geminating ascospores and vegetative hyphae were observed though light microscopy and scanning microscope. During seven weeks of cultivation period, the development process of apical fertile part of stromata can be separated by the development stage of perithecia. To understand the developing process of fruit body at the transcript level, investigating process of distinct gene expression according to cultural condition and developmental stage was discussed.

The purpose of this study was to investigate the influence of handicraft activities on hand promptness and grasp in the elderly. Subjects were comprised of 14 senior citizens between the ages of 70-85, with 7 subjects in the experiment group and 7 in the control group. Subjects in the experiment group practiced various handicrafts twice a day, while those in the control group did not participate in any special activity. The Jebsen Taylor Hand Function Test was used to evaluate the results, while a dynamometer and pinch gauge were used to measure hand promptness and grasp. The 7 senior citizens in the experiment group were able to increase their hand promptness and grasping skills. Conclusively, handicrafts can help improve hand promptness and grasp in the elderly. Furthermore, the development and improvement of such skills can have a positive influence on the daily lives of senior citizens. Such skills are expected to improve the overall neuro-function in the elderly population.