• Asian-Australasian Journal of Animal Sciences
• /
• 제19권11호
• /
• pp.1665-1670
• /
• 2006

The objective of this research was to provide the characterization and method for producing anti-E. coli O157:H7 antibodies in egg-laying hens and to determine if the antibody can restrain the proliferation of E. coli O157:H7 in-vitro. Selected antigenic fractions (whole cell, outer membrane protein and lipopolysaccharide (LPS)) from E. coli O157:H7 were injected to hens in order to produce anti-E. coli O157:H7 antibodies. The immune response and the egg yolk antibodies of laying hens against the whole cell, outer membrane protein and LPS antigens were monitored by ELISA. The level of antibodies against whole cell antigen monitored through ELISA sharply increased after the initial immunization, and it was found to be maximum on day 49 however, the level was maintained up to day 70. Antibodies (5 mg/ml) directed against the whole cell inhibited E. coli proliferation 10-13 times more than outer membrane protein or LPS. The antibody response against the whole cell antigens appeared to have higher activity in restraining the proliferation of E. coli O157:H7 than antibody against outer membrane protein or LPS. Results reflected that increasing the IgY's in the egg yolk could prevent greater economic losses due to human and animal health from pathogenic bacteria i.e. E. coli O157:H7.

• 대한임상검사과학회지
• /
• 제39권1호
• /
• pp.1-6
• /
• 2007

Cervical cancer is one of the most prevalent cancers developed in women worldwide, and human papillomavirus(HPV) type 16 is the most common agent linked to human cerivical carcinoma. Viral oncogenes E6 and E7 are selectively retained and expressed in carcinoma cells infected with human papillomavirus type 16 and cooperate with each other in the immortalization and transformation of primary keratinocytes. Because the HPV oncogenesis mechanism was not completely solved, more thorough studies are required. ln the present study, we investigated the telomere independent role of telomerase in HPV oncogenesis, we constructed the E6 mutant, E7, E6/E7 and hTERT over-expressed stable cells with a telomerase negative cell line, SW13. Expressions of inserted genes were measured by RT-PCR. E6, E7 and hTERT genes were well expressed in each cell lines when compared with the control groups. By analyzing the cell morphology under the microscope, hTERT clone size was a smaller than the mock control but oncogene expressed clones had a slightly lengthened marginal region. In addition, hTERT cells also has a tendency of brief dividing time compared to the mock control. To determine whether telomerase activity was associated with a HPV oncogenesis by oncoprotein expression, we performed the PCR based TRAP assay and a Northern blot analysis. In TRAP assay data, telomerase activities in hTERT and oncogene clones increased compared to the mock control. In addition, SW13/E6/E7 cells showed an extremely increased activity compared to the other clones. Induced hTERT mRNA by E6/E7 wasn't, however, detected in Northern blotting. In conclusion, these findings suggest that telomerase activity is closely associated with the HPV oncogenesis and E6/E7 co-expression is a most important factor of telomerase activity.

• 한국수산과학회지
• /
• 제24권5호
• /
• pp.327-339
• /
• 1991

줄새우아재비, Palaemon serrifer의 뇌와 흉부신경절에 분포하는 신경분비세포와 생식소발달과의 관련 기능을 알아보고자 생식소발달의 조직학적 변화를 조사하여 생식년주기를 밝히고, 뇌와 흉부신경절에 분포하는 신경분비세포를 분류하여 분비활성변화를 조사하였으며, 생식소발달에 따른 이들 분비세포들의 활성변화를 연구하였다. 1. 줄새우아재비, Palaemon serrifer는 수컷의 경우 1월부터, 암컷은 3월부터 생식소가 성장하기 시작하는 성장기를 거쳐, 성숙기, 완숙 및 산란기, 퇴화 및 휴지기의 연속된 생식연주기를 가지며, 주산란기 는 7-8월이었다. 2. 신경분비세포로서 뇌에서는 A-, B-그리고 E-cell이 흉부신경절에서는 A-, A'- 그리고 B-cell이 구분되었으며 A-와 A'-cell은 크기가 $80-90{\mu}m$로 가장 큰 세포였고 B-cell은 $30-40{\mu}m$의 크기였으며, E-cell은 $10-15{\mu}m$ 크기로 미세한 세포였다. 3. 활성중인 A-와 B-cell은 CHP와 AF에, 그리고 B-cell은 AF에만 양성반응을 나타냈었고, A-cell은 분비과립이 축색으로 이동하는 것 외에 주변방출(peripheral discharge)을 나타냈다. 4. 신경분비세포의 활성변화를 생식소발달상태와 연관하여 볼 때 난소의 성장과 성숙시기에는 E-cell, 배란시기에는 A-cell의 분비활성이 강했고, 정소의 성장시기에는 E-cell, 정자의 변태 및 방정시기에는 A-cell의 강한 분비활성이 관찰되었다.

• Journal of Microbiology and Biotechnology
• /
• 제20권10호
• /
• pp.1463-1470
• /
• 2010

E. coli has long been widely used as a host system for the manufacture of recombinant proteins intended for human therapeutic use. When considering the impurities to be eliminated during the downstream process, residual host cell DNA is a major safety concern. The presence of residual E. coli host cell DNA in the final products is typically determined using a conventional slot blot hybridization assay or total DNA Threshold assay. However, both the former and latter methods are time consuming, expensive, and relatively insensitive. This study thus attempted to develop a more sensitive real-time PCR assay for the specific detection of residual E. coli DNA. This novel method was then compared with the slot blot hybridization assay and total DNA Threshold assay in order to determine its effectiveness and overall capabilities. The novel approach involved the selection of a specific primer pair for amplification of the E. coli 16S rRNA gene in an effort to improve sensitivity, whereas the E. coli host cell DNA quantification took place through the use of SYBR Green I. The detection limit of the real-time PCR assay, under these optimized conditions, was calculated to be 0.042 pg genomic DNA, which was much higher than those of both the slot blot hybridization assay and total DNA Threshold assay, where the detection limits were 2.42 and 3.73 pg genomic DNA, respectively. Hence, the real-time PCR assay can be said to be more reproducible, more accurate, and more precise than either the slot blot hybridization assay or total DNA Threshold assay. The real-time PCR assay may thus be a promising new tool for the quantitative detection and clearance validation of residual E. coli host cell DNA during the manufacturingprocess for recombinant therapeutics.

• ALGAE
• /
• 제31권3호
• /
• pp.219-230
• /
• 2016

The Euglena deses group are common freshwater species composed of E. adhaerens, E. carterae, E. deses, E. mutabilis, and E. satelles. These species are characterized by elongated cylindrical worm-like cell bodies and numerous discoid chloroplasts with a naked pyrenoid. To understand the cryptic diversity, species delimitation and phylogenetic relationships among members of the group, we analyzed morphological data (light and scanning electron microscopy) and molecular data (nuclear small subunit [SSU] and large subunit [LSU] rDNAs and plastid SSU and LSU rDNAs). Bayesian and maximum likelihood analyses based on the combined four-gene dataset resulted in a tree consisting of two major clades within the group. The first clade was composed of two subclades: the E. mutabilis subclade, and the E. satelles, E. carterae, and E. adhaerens subclade. The E. mutabilis subclade was characterized by a lateral canal opening at the anterior end and a single pellicular stria, whereas the E. satelles, E. carterae, and E. adhaerens subclade was characterized by an apical canal opening at the anterior end of the cell and double pellicular striae. The second clade consisted of 20 strains of E. deses, characterizing by a subapical canal opening at the anterior end and double pellicular striae, but they showed cell size variation and high genetic diversity. Species boundaries were tested using a Bayesian multi-locus species delimitation method, resulting in the recognition of five cryptic species within E. deses clade.

• Archives of Pharmacal Research
• /
• 제17권6호
• /
• pp.480-482
• /
• 1994

A number of 5-substituted pyrimidine acyclic nucleosides were synthesized and tested for invitor cytotoxicity against four cell lines (j-82 cell, p-388 cell, FM-3A cell and U-938 cell lines). Synthesis of 1-cyanomethyl-5-substituted pyrimidines (1a-e) and 1-(4-cyanobutyl)-5-substituted pyrimidines (2a-e) was acomplished from the series of alkylation reactions ofl 5-substituted uracils with the corresponding chloacetonitrile and 5-chlorovaleronitile in DMSO under $50^{\circ}C$ temperature. These 5-substituted pyrimidine acylic nucleosides (1a-e and 2a-e) exhibited moderate to significant acitivity aginst four cell lines.

• Environmental Analysis Health and Toxicology
• /
• 제29권
• /
• pp.10.1-10.6
• /
• 2014

Objectives Cigarette smoking had been recorded as the main cause of impaired endothelium-dependent vasodilation in smokers by reducing nitric oxide (NO), a production of endothelial nitric oxide synthase (eNOS). However, the mechanism of NO impairment via eNOS activity is unclear until now. In this study, cell passage is suggested to be a relevant factor to eNOS expression under cigarette smoking stress. Methods Bovine aortic endothelial cells (BAECs) were chosen as the research subject with passages ranking from 6 to 9 (6P to 9P). After exposure of cigarette smoking extract (CSE) solution, MTT assay and Western blot method were performed to check the cell viability as well as eNOS protein concentration. In these experiments, four concentrations of CSE at 0.5, 1, 2, and 4% were selected for treatment. Results Our results showed that cells almost died at 4% of CSE. Besides, eNOS protein mass had a linear decrease under the increase of CSE concentration. In addition, the effect of CSE on eNOS expression was dissimilar between different passages. Conclusions This study indicated that CSE had effect on both cell viability and eNOS expression. Besides, a reduction in protein mass was matched with the decrease of cell viability due to CSE tress. Last but not least, the response of eNOS protein to different concentration of CSE at different passages was disparate, making the hypothesis about cell passage related inhibition of eNOS caused by CSE solution.

• 대한의생명과학회지
• /
• 제12권4호
• /
• pp.399-403
• /
• 2006

Cervical cancer is one of the most prevalent cancers developed in women worldwide, and human papillomavirus (HPV) type 16 is the most common agent linked to human cerivical carcinoma. Viral oncogenes E6 and E7 are selectively ratined and expressed in carcinoma cells infected with human papillomavirus type 16 and cooperated with each other in immortalization and transformation of primary keratinocytes. Because of HPV oncogenesis mechanism was not completely solved, the more studies be required thoroughly. In the present study, to investigate the telomere independent role of telomerase in HPV oncogenesis, we constructed the E6 mutant, E7, E6/E7 and hTERT over-expressed stable cells with a telomerase negative cell line, SW13. Expressions of Inserted genes were measured by RT-PCR. E6, E7 and hTERT genes were well expressed in each cell lines comparing with the control groups. By analyzing the cell morphology under the microscope, hTERT clone size was a more smaller than the mock control but oncogene expressed clones were slightly lengthened the marginal region. In addition, hTERT cells has also, a tendency of brief dividing time compared to the mock control. To determine whether telomerase activity associated with a HPV oncogenesis by oncoprotein expression, we performed the PCR based TRAP assay and Northern blot analysis. In TRAP assay data, telomerase activities in hTERT and oncogene clones were more increased than the mock control. In addition, SW13/ E6/E7 cells appeared a extremely increased activity than any other clones. Induced TERT mRNA by E6/E7 wasn't, however, detected in Northern blotting. In conclusion, these findings suggest that telomerase activity closely associated the HPV oncogenesis and E6/E7 co-expression is a most important factor of telomerase activity.

• IMMUNE NETWORK
• /
• 제4권1호
• /
• pp.44-52
• /
• 2004

Background: As a potent antigen presenting cell and a powerful inducer of antigen specific immunity, dendritic cells (DCs) are being considered as a promising anti-tumor therapeutic module. The expected therapeutic effect of DCs in renal cell carcinoma was tested in the mouse model. Established late-stage tumor therapeutic (E-T) and minimal residual disease (MRD) model was considered in the in vivo experiments. Methods: Syngeneic renal cell carcinoma cells (RENCA) were inoculated either subcutaneously (E-T) or intravenously (MRD) into the Balb/c mouse. Tumor cell lysate pulsed-DCs were injected twice in two weeks. Intraperitoneal DC injection was started 3 week (E-T model) or one day (MRD model) after tumor cell inoculation. Two weeks after the final DC injection, the tumor growth and the systemic immunity were observed. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 for 7 days and pulsed with RENCA cell lysate for 18 hrs. Results: Compared to the saline treated group, tumor growth (E-T model) or formation (MRD model) was suppressed in pulsed-DC treated group. RENCA specific lymphocyte proliferation was observed in the RENCA tumor-bearing mice treated with pulsed-DCs. Primary cytotoxic T cell activity against RENCA cells was increased in pulsed-DC treated group. Conclusion: The data suggest the possible anti-tumor effect of cultured DCs in established or minimal residual disease/metastasis state of renal cell carcinoma. Systemic tumor specific immunity including cytotoxic T cell activity was modulated also in pulsed-DC treated group.

• 한국통신학회논문지
• /
• 제36권3A호
• /
• pp.240-249
• /
• 2011

본 논문은 펨토셀시스템(femtocell system)의 단말(mobile station)이 핸드오버(handover)하기 전에 이웃한 기지국(neighbor station)의 정보를 공유하는 셀 관리(cell management) 기법과 핸드오버 방법에 대해 제안한다. 펨토셀시스템에서는 셀관리를 위해 수많은 펨토셀의 정보를 MOB_NBR-ADV메시지에 포함하여 방송하는데, 단말과 기지국에서 MOB_NBR-ADV메시지를 처리할 때 일시적인 부하가 발생된다. 따라서 효율적인 동작과 핸드오버 성공률 개선을 위해 MOB_NBR-ADV메시지의 구성 및 전송방법을 제안한다. 또한, 셀 간 중첩된 영역에서 핸드오버의 핑퐁(ping-pong) 현상을 막기 위해 히스테리시스 범위(hysteresis margin)를 이용한다. 그러나 수많은 펨토셀이 도입된 경우 기존의 히스테리시스범위 만으로 핑퐁현상을 막을 수 없다. 본 논문에서는 중첩된 셀의 개수가 증가하는 경우 히스테리시스범위가 커지는 방법과, 차등적으로 셀을 관리하여 불필요한 핸드오버의 횟수를 최소화하는 방법 등을 제안하였다. 컴퓨터 모의실험을 통해 성능 검증한 결과, 제안한 방식이 기존의 펨토셀 시스템의 셀 관리 및 핸드오버 방식에 비해 성능을 개선시키는 것을 확인하였다.