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Development and Characterization of Hyperglycosylated Recombinant Human Erythropoietin (HGEPO)

  • JarGal, Naidansuren;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.77-83
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    • 2009
  • Erythropoietin (EPO), a glycoprotein hormone produced from primarily cells of the peritubular capillary endothelium of the kidney, is responsible for the regulation of red blood cell production. We have been investigating the roles of glycosylation site added in the biosynthesis and function of recombinant protein. We constructed three EPO mutants ($\Delta$69, $\Delta$105 and $\Delta$69,105), containing an additional oligosaccharide chains. EPOWT and EPO$\Delta$69 were effectively expressed in transient and stably transfected CHO-K1 cell lines. But, it wasn't detected any protein in the culture medium of EPO$\Delta$105 and EPO$\Delta$69,105 mutants. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure the cytokine dependency and in vitro bioactivity of rec-hEPO. MTT assay values were increased by survival of F36E cells at 24h. To analysis biological activity in vivo, two groups of ICR-mice (7 weeks old) were injected subcutaneously with 10 IU per mice of rec-hEPO molecules on days 0 and 2. Red blood cell and hematocrit values were measured on 6 days after the first injection. The hematocrit values were remarkably increased in all treatment groups. The pharmacokinetic analysis was also affected in the mice injected with rec-hEPO molecules 2.5 IU by tail intravenous. Protein samples were detected by Western blotting. An EPO$\Delta$69 protein migrated as a broad band with an average apparent molecular and detected slightly high band. Enzymatic N-deglycosylation resulted in narrow band and was the same molecular size. The biological activity of EPO$\Delta$69 was enhanced to compare with wt-hEPO. The half-life was longer than wt-hEPO. The results suggest that hyperglycosyalted recombinant human erythropoietin (EPO$\Delta$69) may have important biological and therapeutic good points.

Clinico-pathology of Lung Cancer in a Regional Cancer Center in Northeastern India

  • Mandal, Sanjeet Kumar;Singh, Thaudem Tomcha;Sharma, Takhenchangbam Dhaneshor;Amrithalingam, Venkatesan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7277-7281
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    • 2013
  • Background: Globally, there have been important changes in trends amongst gender, histology and smoking patterns of lung cancer cases. Materials and Methods: This retrospective study was conducted on 466 patients with lung cancer who were registered in Regional Cancer Center, Regional Institute of Medical Sciences, Manipur from January 2008 to December 2012. Results: Most were more than 60 years of age (67.8%) with a male: female ratio of 1.09:1. Some 78.8% of patients were chronic smokers with male smoker to female smoker ratio of 1.43:1. Consumption of alcohol was found in 29.4%, both smoking and alcohol in 27.5%, betel nut chewing in 37.9% and tobacco chewing in 25.3%. A history of tuberculosis was present in 16.3% of patients. The most frequent symptom was coughing (36.6%) and most common radiological presentation was a mass lesion (70%). Most of the patients had primary lung cancer in the right lung (60.3%). The most common histological subtype was squamous cell carcinoma (49.1%), also in the 40-60 year age group (45.9%), more than 60 year age group (51.6%), males (58.1%) and females (41.8%). As many as 91.9% of squamous cell carcinoma patients had a history of smoking. About 32.5% of patients had distant metastasis at presentation with brain (23.8%) and positive malignant cells in pleural effusions (23.1%) as common sites. The majority of patients were in stage III (34.4%), stage IV (32.5%) and stage II (30.2%). Conclusions: Our analysis suggests that the gender gap has been narrowed such that about half of the patients diagnosed with lung cancer are women in this part of India. This alarming rise in female incidence is mainly attributed to an increased smoking pattern. Squamous cell carcinoma still remains the commonest histological subtype. Most of the patients were elderly aged and presented at locally or distantly advanced stages.

Latent Transforming Growth Factor-beta1 Functionalised Electrospun Scaffolds Promote Human Cartilage Differentiation: Towards an Engineered Cartilage Construct

  • Lim, Erh-Hsuin;Sardinha, Jose Paulo;Myers, Simon;Stevens, Molly
    • Archives of Plastic Surgery
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    • v.40 no.6
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    • pp.676-686
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    • 2013
  • Background To overcome the potential drawbacks of a short half-life and dose-related adverse effects of using active transforming growth factor-beta 1 for cartilage engineering, a cell-mediated latent growth factor activation strategy was developed incorporating latent transforming growth factor-${\beta}$1 (LTGF) into an electrospun poly(L-lactide) scaffold. Methods The electrospun scaffold was surface modified with NH3 plasma and biofunctionalised with LTGF to produce both random and orientated biofunctionalised electrospun scaffolds. Scaffold surface chemical analysis and growth factor bioavailability assays were performed. In vitro biocompatibility and human nasal chondrocyte gene expression with these biofunctionalised electrospun scaffold templates were assessed. In vivo chondrogenic activity and chondrocyte gene expression were evaluated in athymic rats. Results Chemical analysis demonstrated that LTGF anchored to the scaffolds was available for enzymatic, chemical and cell activation. The biofunctionalised scaffolds were non-toxic. Gene expression suggested chondrocyte re-differentiation after 14 days in culture. By 6 weeks, the implanted biofunctionalised scaffolds had induced highly passaged chondrocytes to re-express Col2A1 and produce type II collagen. Conclusions We have demonstrated a proof of concept for cell-mediated activation of anchored growth factors using a novel biofunctionalised scaffold in cartilage engineering. This presents a platform for development of protein delivery systems and for tissue engineering.

Anticancer Effect of COX-2 Inhibitor DuP-697 Alone and in Combination with Tyrosine Kinase Inhibitor (E7080) on Colon Cancer Cell Lines

  • Altun, Ahmet;Turgut, Nergiz Hacer;Kaya, Tijen Temiz
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3113-3121
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    • 2014
  • Colorectal cancer remains one of the most common types of cancer and a leading cause of cancer death worldwide. In this study, we aimed to investigate effects of DuP-697, an irreversible selective inhibitor of COX-2 on colorectal cancer cells alone and in combination with a promising new multi-targeted kinase inhibitor E7080. The HT29 colorectal cancer cell line was used. Real time cell analysis (xCELLigence system) was conducted to determine effects on colorectal cell proliferation, angiogenesis was assessed with a chorioallantoic membrane model and apoptosis was determined with annexin V staining. We found that DuP-697 alone exerted antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells. For the antiproliferative effect the half maximum inhibition concentration ($IC_{50}$) was $4.28{\times}10^{-8}mol/L$. Antiangiogenic scores were 1.2, 0.8 and 0.5 for 100, 10 and 1 nmol/L DuP-697 concentrations, respectively. We detected apoptosis in 52% of HT29 colorectal cancer cells after administration of 100 nmol/L DuP-697. Also in combination with the thyrosine kinase inhibitor E7080 strong antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells were observed. This study indicates that DuP-697 may be a promising agent in the treatment of colorectal cancer. Additionally the increased effects observed in the combination with thyrosine kinase inhibitor give the possibility to use lower doses of DuP-697 and E7080 which can avoid and/or minimize side effects.

담배세초현탁배양을 이용한 human granulocyte-macrophage colony stimulating factor의 생산에서 배지 성분이 미치는 영향

  • Lee, Gi-Yong;Lee, Sang-Yun;Myeong, Hyeon-Jong;No, Yun-Suk;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.325-328
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    • 2002
  • Production of human granulocyte-macrophage colony stimulating factor (hGM-CSF) by Nicotiana tabacum cell suspension culture was studied in Murashige and Skoog (MS) medium with sucrose as a carbon source, ammonium nitrate and potassium nitrate as nitrogen sources, potassium dihydrogen phosphate and sodium dihydrogen phosphate hydrate as phosphate sources, respectively. Optimum concentrations for carbon, nitrogen, phosphate was determined to enhance the production of hGM-CSF. Cell growth was better at high initial sucrose concentration (60 g/L), high initial nitrogen concentration (121.04 mM). Maximum cell density (18.28 g/L) was obtained at 60 g/L of sucrose after 14 days. Cell growth was not so good at low initial sucrose concentration 00 g/L), but the highest hGM-CSF production was obtained at the latter half of exponential phase. hGM-CSF production increased about 3 fold at initial phosphate concentration of 4.96 nM

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The Effects of Taking Elective Chemistry II Courses in High School on Understanding Concepts of Electrochemistry in General Chemistry: Focusing on Chemical Cell (고등학교 화학II 선택과목 이수가 대학 일반화학의 전기화학 관련 개념의 이해에 미치는 영향: 화학전지를 중심으로)

  • Yang, Hye-Ran;Lee, Sang Kwon
    • Journal of the Korean Chemical Society
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    • v.61 no.1
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    • pp.34-44
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    • 2017
  • The purpose of this study was to analyze the conception type change and to investigate the effect of understanding on concepts in electrochemistry after general chemistry lessons. The significant differences in concept understanding of electrochemistry were shown in both groups. Statistically significant gains in both groups were as shown in distinguishing the chemical cell, in identifying the anode and cathode, and in understanding current formation and flow, while, significant achievements in understanding the role of the salt bridge, and the need for a standard half-cell were not found. Taking elective chemistry II in high school had an effect on understanding related concepts of electrochemistry in general chemistry lessons. It was shown that many freshmen had difficulties in understanding exact related concepts in several kinds after general chemistry lessons. In order to solve these problems, it is necessary to teach contents of the basic concepts in electrochemistry exactly and to hold supplementary lessons.

Enhancing Electricity Generation Using a Laccase-Based Microbial Fuel Cell with Yeast Galactomyces reessii on the Cathode

  • Chaijak, Pimprapa;Sukkasem, Chontisa;Lertworapreecha, Monthon;Boonsawang, Piyarat;Wijasika, Sutthida;Sato, Chikashi
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1360-1366
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    • 2018
  • The fungi associated with termites secrete enzymes such as laccase (multi-copper oxidase) that can degrade extracellular wood matrix. Laccase uses molecular oxygen as an electron acceptor to catalyze the degradation of organic compounds. Owing to its ability to transfer electrons from the cathodic electrode to molecular oxygen, laccase has the potential to be a biocatalyst on the surface of the cathodic electrode of a microbial fuel cell (MFC). In this study, a two-chamber MFC using the laccase-producing fungus Galactomyces reessii was investigated. The fungus cultured on coconut coir was placed in the cathode chamber, while an anaerobic microbial community was maintained in the anode chamber fed by industrial rubber wastewater and supplemented by sulfate and a pH buffer. The laccase-based biocathode MFC (lbMFC) produced the maximum open circuit voltage of 250 mV, output voltage of 145 mV (with a $1,000{\Omega}$ resistor), power density of $59mW/m^2$, and current density of $278mA/m^2$, and a 70% increase in half-cell potential. This study demonstrated the capability of laccase-producing yeast Galactomyces reessii as a biocatalyst on the cathode of the two-chamber lbMFC.

Increase in apoptotic effect of Panax ginseng by microwave processing in human prostate cancer cells: in vitro and in vivo studies

  • Park, Jun Yeon;Choi, Pilju;Kim, Ho-kyong;Kang, Ki Sung;Ham, Jungyeob
    • Journal of Ginseng Research
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    • v.40 no.1
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    • pp.62-67
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    • 2016
  • Background: Ginseng, which is widely used in functional foods and as an herbal medicine, has been reported to reduce the proliferation of prostate cancer cells by mechanisms that are not yet fully understood. Methods: This study was designed to investigate the changes in ginsenoside content in ginseng after treatment with a microwave-irradiation thermal process and to verify the anticancer effects of the extracts. To confirm the anticancer effect of microwave-irradiated processed ginseng (MG), it was tested in three human prostate cancer cell lines (DU145, LNCaP, and PC-3 cells). Involvements of apoptosis and autophagy were assessed using Western blotting. Results: After microwave treatment, the content of ginsenosides Rg1, Re, Rb1, Rc, Rb2, and Rd in the extracts decreased, whereas the content of ginsenosides 20(S)-Rg3, 20(R)-Rg3, Rk1, and Rg5 increased. Antiproliferation results for the human cancer cell lines treated with ginseng extracts indicate that PC-3 cells treated with MG showed the highest activity with an half maximal inhibitory concentration of $48{\mu}g/mL$. We also showed that MG suppresses the growth of human prostate cancer cell xenografts in athymic nude mice as an in vivo model. This growth suppression by MG is associated with the inductions of cell death and autophagy. Conclusion: Therefore, heat processing by microwave irradiation is a useful method to enhance the anticancer effect of ginseng by increasing the content of ginsenosides Rg3, Rg5, and Rk1.

Enhancement on the Charge-discharge Property of Carbon Anode by the Addition of Metal Oxides in Li-ion Secondary Batteries (금속산화물 첨가방법에 의한 리튬이차전지 부극재료의 충방전 특성 개선)

  • 김정식
    • Journal of the Korean Ceramic Society
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    • v.40 no.11
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    • pp.1085-1089
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    • 2003
  • In the present study effects of SnO$_2$-impregnation on the cell performance of Mesocarbon Microbeads (MCMB) electrode in the Li-ion battery have been investigated. Sn element was impreganted into MCMB powders by the chemical titration, and then post annealed at 250$^{\circ}C$ for 1 h in ambient atmosphere to be transformed as tin-oxide. From the measurement for the cell performance with the half cell in which the SnO$_2$-impregnated MCMB was used as an anode, the SnO$_2$-impregnated MCMB showed higher charge/discharge capacities, higher reversible specific charge capacity and better cycleability than a raw MCMB. As the amount of impregnated SnO$_2$ increased, both reversible and irreversible capacities increased.

Optical and Electrical Properties of InAs Sub-Monolayer Quantum Dot Solar Cell

  • Han, Im-Sik;Park, Dong-U;No, Sam-Gyu;Kim, Jong-Su;Kim, Jin-Su;Kim, Jun-O
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.196.2-196.2
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    • 2013
  • 본 연구에서는 분자선 에피택시 (MBE)법으로 성장된 InAs submonolayer quantum dot (SML-QD)을 태양전지에 응용하여 광학 및 전기적 특성을 평가하였다. 본 연구에서 사용된 양자점 태양전지(quantum dot solar cell, QDSC)의 구조는 n+-GaAs 기판 위에 n+-GaAs buffer와 n-GaAs base layer를 차례로 성장 한 후, 활성영역에 InAs/InGaAs SML-QD와 n-GaAs spacer layer를 8주기 형성하였다. 그 위에 p+-GaAs emitter, p+-AlGaAs window layer를 성장하고 ohmic contact을 위하여 p+-GaAs 를 성장하였다. SML-QD 구조의 두께는 0.3 ML 이며, 이때 SML-QD의 적층수를 4 stacks 으로 고정하였다. SML-QD 와의 비교를 위하여 2.0 ML크기의 InAs자발 형성 양자점 태양전지(SK-QDSC)과 GaAs 단일 접합 태양전지 (reference-SC)를 동일한 성장조건에서 제작하였다. PL 측정 결과, 300 K에서 SML-QD의 발광 피크는 SK-QD 보다 고에너지에서 나타나는데(1.349 eV), 이것은 SML-QD가 SK-QD보다 작은 크기를 가지기 때문으로 사료된다. SML-QD는 single peak를 보이는 반면, SK-QD는 dual peaks (1.112 / 1.056 eV)을 확인하였다. SML-QD의 반치폭(full width at half maximum, FWHM)이 SK-QD에 비하여 작은 것으로 보아 SML-QD가 SK-QD보다 양자점 크기 분포의 균일도가 높은 것으로 해석된다. Illumination I-V 측정 결과, SML-QDSC의 개방 전압(VOC) 과 단락전류밀도(JSC)는 SK-QDSC의 값과 비교해 보면, 각각 47 mV와 0.88 mA/cm2만큼 증가하였다. 이는 SK-QD보다 상대적으로 작은 크기를 가진 SML-QD로 인해 VOC가 증가되었으며, SML-QD가 SK-QD 보다 태양광을 흡수할 수 있는 영역이 비교적 적지만, QD내에 존재하는 energy level에서 탈출 할 수 있는 확률이 더 높음으로써 JSC가 증가한 것으로 분석 된다.

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