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Sample records for competent cell preparation

  1. Competency-Based Reading Preparation for Secondary Teachers.

    Science.gov (United States)

    Getz, Howard G.; Kennedy, Larry D.

    The reading preparation program at Illinois State University represents a fundamental shift in the instructional focus for preservice preparation of secondary level teachers. The competency-based program in reading is largely self-directed and self-paced. Large groups of students are assigned to an instructional team, and the faculty provides…

  2. 高效JM109感受态细胞制备及转化条件的优化%Preparation and Transformation Conditions for Efficient JM109 Competent Cells

    Institute of Scientific and Technical Information of China (English)

    许彤; 刘钦松; 张丛丛; 刘孟刚

    2012-01-01

    Competent cells of Escherichia coli JM109 were prepared by different transformation liquid, and pUC18 was transformed into the competent cells. The effects of different growth state of bacteria, transformation liquid, heat shock time and culture medium after heat shock on transformation efficiency were studied. The results showed that the highest transformation rate could be obtained by preparing the competent cells by TB transformation liquid when bacteria A600nm was 0.705, heat shocked at 42℃ for 45 s and then culturing in SOC medium. The transformation efficiency could reach to 8.59×108 CFU/μg plasmid.%采用不同的转化液制备大肠杆菌(Escherichia coli)JM109感受态细胞,并进行pUC18质粒的转化,考察细菌生长状态、转化液、转化的热激时间和转化后所用培养基对转化率的影响.结果表明,菌液A600nm为0.705时采用TB转化液制备感受态细胞,在42℃热激45 s,转化后采用SOC培养基振荡培养,转化效率最高,可达8.59×108 CFU/μg质粒.

  3. 76 FR 72619 - User Fee To Take the Registered Tax Return Preparer Competency Examination

    Science.gov (United States)

    2011-11-25

    ... Competency Examination AGENCY: Internal Revenue Service (IRS), Treasury. ACTION: Final regulations. SUMMARY... a ] user fee for individuals to take the registered tax return preparer competency examination. The final regulations affect individuals who take the registered tax return preparer competency...

  4. Professional preparation in physical education: changing labor market and competence

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    Go Tani

    2013-09-01

    Full Text Available Professional preparation is indeed a complex and dynamic process because it involves a number of interacting elements, which change in time. The objective of the present essay is to analyze the professional preparation in physical education, with the focus on the relation between the very dynamic labor market and the required competence of the professionals to deal with the associated demands. There is no doubt that the professional preparation must not aim to train professionals to merely repeat means for solving practical problems, but professionals with the capacity to repeat the process of solving problems. Consequently, professional preparation programs need to be formative instead of informative and prepare professionals capable of using scientific thinking and method to solve practical problems of intervention.

  5. Building Emotional Competence: A Strategy for Disaster Preparation and Recovery

    Science.gov (United States)

    West, Nancie Tonner; Albrecht, Kay

    2007-01-01

    Emotional competency is defined as developed ability and skills in the areas of self-awareness, self-regulation, social awareness, and relationship management. These skills are nurtured, developed, and practiced until they become competencies and serve as a resource when a tragic event occurs. They are relatively undeveloped in very young children…

  6. Diversity Competencies within Early Childhood Teacher Preparation: Innovative Practices and Future Directions

    Science.gov (United States)

    Lim, Chih-Ing; Able-Boone, Harriet

    2005-01-01

    With classrooms becoming increasingly diverse due to children's various cultural backgrounds and varying abilities, early childhood teacher education programs face the challenge of how best to prepare the workforce. Various initiatives have been implemented in teacher preparation programs to prepare early childhood educators to become competent in…

  7. Developing palliative care competencies for the education of entry level baccalaureate prepared Canadian nurses.

    Science.gov (United States)

    Jacono, Brenda; Young, Lynne; Baker, Cynthia; Richardson, Holly R L; Cable-Williams, Beryl; Jewers, Heather; Lavoie, Mireille; Librach, Larry; Bidgood, Darcee; Mitchell, Mitzi Grace

    2011-08-15

    Educational preparation of health professionals for Palliative and End of Life Care (PEOLC) is inadequate, and nurses are no exception. In 2004, the Canadian Association of Schools of Nursing struck a Task Force to develop PEOLC competencies to address this issue. The development of national PEOLC nursing competencies involved a multi-step, emergent, interactive, and iterative process. An overarching principle guiding this process was building national consensus about the essential PEOLC specific competencies for nurses among experts in this field while simultaneously generating, revising, and refining them. There have been three stages in this iterative, multi-step process: 1) Generating a preliminary set of competencies, 2) Building a national consensus among educators and experts in the field on PEOLC specific competencies for nurses, and 3) Refining the consensus based competencies for curriculum development. Ongoing follow up work for this project is focusing on the integration of these competencies into nursing curricula.

  8. A Competency-Based Approach to the Master's Degree Preparation of Higher Education Professionals

    Science.gov (United States)

    Ott, Molly; Baca, Evelyn; Cisneros, Jesus; Bates, Evan

    2015-01-01

    The purpose of this manuscript is to describe a competency-based approach to designing and assessing master's level professional preparation programs in the field of higher education administration. Given the absence of a universal set of competencies defined for HEA master's degree programs, the authors draw from the CAHEP (2010) and Wright…

  9. Standardized Competencies for Parenteral Nutrition Order Review and Parenteral Nutrition Preparation, Including Compounding: The ASPEN Model.

    Science.gov (United States)

    Boullata, Joseph I; Holcombe, Beverly; Sacks, Gordon; Gervasio, Jane; Adams, Stephen C; Christensen, Michael; Durfee, Sharon; Ayers, Phil; Marshall, Neil; Guenter, Peggi

    2016-08-01

    Parenteral nutrition (PN) is a high-alert medication with a complex drug use process. Key steps in the process include the review of each PN prescription followed by the preparation of the formulation. The preparation step includes compounding the PN or activating a standardized commercially available PN product. The verification and review, as well as preparation of this complex therapy, require competency that may be determined by using a standardized process for pharmacists and for pharmacy technicians involved with PN. An American Society for Parenteral and Enteral Nutrition (ASPEN) standardized model for PN order review and PN preparation competencies is proposed based on a competency framework, the ASPEN-published interdisciplinary core competencies, safe practice recommendations, and clinical guidelines, and is intended for institutions and agencies to use with their staff.

  10. Forming of communicative competence as condition of professional preparation of future teachers of physical culture

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    Samsutina NM.

    2010-02-01

    Full Text Available The modern state and necessity of realization of forming communicative competence of future teachers of physical culture is found out in the process of professional preparation. 294 students took part in an experiment. Rotined expedience of realization of forming of communicative competence of future teachers of physical culture. The questionnaire of students of higher educational establishments is conducted. The level of formed of communicative competence for students remains at low level. It needs strengthening of attention to perfection of process of professional preparation of future teachers of physical culture.

  11. Administrators' ratings of competencies needed to prepare preservice teachers for oral deaf education programs.

    Science.gov (United States)

    Lartz, Maribeth N; Litchfield, Sharon K

    Deaf education teacher preparation programs must prepare teachers to staff an increasing number of oral programs. A survey was conducted to determine which competencies administrators of deaf education programs rate as important for teachers in oral programs and to compare ratings of these competencies by oral school administrators to ratings made by administrators of comprehensive deaf and hard of hearing programs. Between the two groups of administrators, six areas of agreement about competencies were found. There were notable differences in the range of ratings between the two groups. These differences were attributed to the roles teachers assume in the two types of programs and the focus of instruction in each type of program.

  12. Perceptions of veterinary faculty members regarding their responsibility and preparation to teach non-technical competencies.

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    Lane, India F; Bogue, E Grady

    2010-01-01

    The development of non-technical competencies has become an important component of veterinary education. In this study, we determined faculty perspectives regarding their perceived involvement and ability in the cultivation of these competencies. A survey was administered to faculty members at five institutions. Respondents were asked whether the competency should be taught in their own courses and how prepared they felt to teach and evaluate the competency. Responses were analyzed by participant institution, gender, terminal degree and year, discipline, rank, and teaching experience. More than 90% of faculty respondents reported a personal responsibility to teach or cultivate critical thinking skills, communication skills, self-development skills, and ethical skills, with more than 85% also agreeing to a role in skills such as interpersonal skills, creativity, and self-management. The lowest percentages were seen for crisis and incident management (64%) and business skills (56%). Perceived preparedness to teach and evaluate these competencies paralleled the preceding findings, especially for the four consensus competencies and self-management. Faculty preparedness was lowest for business skills. Junior faculty were somewhat less likely than others to perceive a responsibility to teach non-technical competencies; however, instructors were more prepared to teach and evaluate business skills than were other faculty. Institutional trends were evident in faculty preparation. Although male faculty and non-DVM faculty tended to report a higher degree of preparedness, few differences reached statistical significance. Faculty perceptions of their responsibility to teach non-technical competencies vary by competency and parallel their perceived preparedness to teach and evaluate them.

  13. Social Work Preparation to Compete in Today's Scientific Marketplace

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    Nurius, Paula S.

    2017-01-01

    As the scientific marketplace rapidly evolves, we must keep revisiting strategic preparation of our doctoral students and early career scholars to be successful innovators in these contexts. As an inherently integrative, change-oriented, community-engaged, and context-sensitive discipline, social work has enormous potential as a value-added…

  14. The Preparation of Highly Motivated and Professionally Competent Teachers in Initial Teacher Education

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    Tang, Sylvia Y. F.; Wong, Angel K. Y.; Cheng, May M. H.

    2015-01-01

    Education systems around the world need to recruit highly motivated individuals to become teachers and prepare professionally competent teacher education graduates to take up these broadened and deepened roles and responsibilities with a deep and lasting engagement to the profession. This article reports on a mixed-methods study that examines…

  15. The Implementation of an Intercultural Competence Syllabus to Prepare Study-Abroad Students for Global Communication

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    Kural, Faruk; Bayyurt, Yasemin

    2016-01-01

    This study presents the outcome of the implementation of a process-oriented model of an intercultural competence (IC) and English as a lingua franca (ELF)-awareness development syllabus to prepare government-sponsored Turkish international sojourners for global communication in English L1 countries. Based on social constructivist research…

  16. The Implementation of an Intercultural Competence Syllabus to Prepare Study-Abroad Students for Global Communication

    Science.gov (United States)

    Kural, Faruk; Bayyurt, Yasemin

    2016-01-01

    This study presents the outcome of the implementation of a process-oriented model of an intercultural competence (IC) and English as a lingua franca (ELF)-awareness development syllabus to prepare government-sponsored Turkish international sojourners for global communication in English L1 countries. Based on social constructivist research…

  17. Dynamized Preparations in Cell Culture

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    Ellanzhiyil Surendran Sunila

    2009-01-01

    Full Text Available Although reports on the efficacy of homeopathic medicines in animal models are limited, there are even fewer reports on the in vitro action of these dynamized preparations. We have evaluated the cytotoxic activity of 30C and 200C potencies of ten dynamized medicines against Dalton's Lymphoma Ascites, Ehrlich's Ascites Carcinoma, lung fibroblast (L929 and Chinese Hamster Ovary (CHO cell lines and compared activity with their mother tinctures during short-term and long-term cell culture. The effect of dynamized medicines to induce apoptosis was also evaluated and we studied how dynamized medicines affected genes expressed during apoptosis. Mother tinctures as well as some dynamized medicines showed significant cytotoxicity to cells during short and long-term incubation. Potentiated alcohol control did not produce any cytotoxicity at concentrations studied. The dynamized medicines were found to inhibit CHO cell colony formation and thymidine uptake in L929 cells and those of Thuja, Hydrastis and Carcinosinum were found to induce apoptosis in DLA cells. Moreover, dynamized Carcinosinum was found to induce the expression of p53 while dynamized Thuja produced characteristic laddering pattern in agarose gel electrophoresis of DNA. These results indicate that dynamized medicines possess cytotoxic as well as apoptosis-inducing properties.

  18. Preparing Globally Competent Teacher Candidates Through CrossCultural Experiential Learning

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    Michael A. Kopish

    2016-11-01

    Full Text Available This manuscript presents findings and implications from a case study of one global educator's attempt to develop globally competent teacher candidates in an elective teacher preparation course. Global Citizenship Education served as the framing paradigm for the course and human experiences of immigrants and refugees served as the milieu for teacher candidates to learn critical inquiry. Teacher candidates also participated in several cross-cultural experiential learning opportunities designed to facilitate the development of global competencies (Longview, 2008 in teacher candidates. Students' reflective journals were analyzed to determine the personal significance of different learning experiences and the extent to which teacher candidates’ perceptions of immigrants and refugees changed as a result of the course content and activities. The findings demonstrate the potential of critical inquiry and cross-cultural experiential learning as transformative teaching practices to develop globally competent teachers.

  19. Revisiting oocyte–somatic cell interactions: in search of novel intrafollicular predictors and regulators of oocyte developmental competence

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    Li, Qinglei; McKenzie, Laurie J.; Matzuk, Martin M.

    2008-01-01

    Prediction and improvement of oocyte competence are two critical issues in assisted reproductive technology to improve infertility therapy. The lack of reliable and objective predictors of oocyte developmental competence for oocyte/embryo selection during in vitro fertilization hampers the effectiveness of this technology. Likewise, the low pregnancy rate resulting from in vitro maturation of human oocytes represents a major obstacle for its clinical application. Oocyte competence is progressively acquired during follicular development, and the oocyte plays a dominant role in regulating granulosa cell functions and maintaining the microenvironment appropriate for the development of its competence. Hence, granulosa cell functions are reflective of oocyte competence, and molecular markers of granulosa cells are potentially reliable predictors of oocyte quality. With the advent of the functional genomics era, the transcriptome of granulosa cells has been extensively characterized. Experimental data supporting granulosa cell markers as predictors of oocyte competence are now emerging in both animal models and humans. Future efforts should focus on integrating granulosa cell genetic markers as parameters for oocyte/embryo selection. Moreover, novel in vitro evidence highlights the effectiveness of exogenous oocyte-secreted factors in promoting oocyte developmental competence in animal models. The challenge in evaluating the effect of oocyte-secreted factors on oocyte quality in a clinical setting is to standardize the various preparations of these recombinant proteins and decipher their complex interactions/cooperativity within the germline-somatic cell regulatory loop. PMID:18996952

  20. Mural granulosa cell gene expression associated with oocyte developmental competence

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    Jiang Jin-Yi

    2010-03-01

    Full Text Available Abstract Background Ovarian follicle development is a complex process. Paracrine interactions between somatic and germ cells are critical for normal follicular development and oocyte maturation. Studies have suggested that the health and function of the granulosa and cumulus cells may be reflective of the health status of the enclosed oocyte. The objective of the present study is to assess, using an in vivo immature rat model, gene expression profile in granulosa cells, which may be linked to the developmental competence of the oocyte. We hypothesized that expression of specific genes in granulosa cells may be correlated with the developmental competence of the oocyte. Methods Immature rats were injected with eCG and 24 h thereafter with anti-eCG antibody to induce follicular atresia or with pre-immune serum to stimulate follicle development. A high percentage (30-50%, normal developmental competence, NDC of oocytes from eCG/pre-immune serum group developed to term after embryo transfer compared to those from eCG/anti-eCG (0%, poor developmental competence, PDC. Gene expression profiles of mural granulosa cells from the above oocyte-collected follicles were assessed by Affymetrix rat whole genome array. Results The result showed that twelve genes were up-regulated, while one gene was down-regulated more than 1.5 folds in the NDC group compared with those in the PDC group. Gene ontology classification showed that the up-regulated genes included lysyl oxidase (Lox and nerve growth factor receptor associated protein 1 (Ngfrap1, which are important in the regulation of protein-lysine 6-oxidase activity, and in apoptosis induction, respectively. The down-regulated genes included glycoprotein-4-beta galactosyltransferase 2 (Ggbt2, which is involved in the regulation of extracellular matrix organization and biogenesis. Conclusions The data in the present study demonstrate a close association between specific gene expression in mural granulosa cells and

  1. A Commentary on Innovation and Emerging Scientific Careers: Is Social Work Prepared to Compete in Today's Scientific Marketplace?

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    Craddock, Jaih B.

    2017-01-01

    The aim of this article is to address some of the questions Dr. Paula S. Nurius presents in her article, "Innovation and Emerging Scientific Careers: Is Social Work Prepared to Compete in Today?s Scientific Marketplace?" Specifically, this article will focus on what we can do to better prepare our emerging research scholars to be…

  2. Replication-competent human adenovirus 11p vectors can propagate in Vero cells

    Energy Technology Data Exchange (ETDEWEB)

    Gokumakulapalle, Madhuri; Mei, Ya-Fang, E-mail: ya-fang.mei@umu.se

    2016-08-15

    The use of continuous cell lines derived from the African green monkey kidney (AGMK) has led to major advances in virus vaccine development. However, to date, these cells have not been used to facilitate the creation of human adenoviruses because most human adenoviruses undergo abortive infections in them. Here, we report the susceptibility of AGMK-derived cells to adenovirus 11p (Ad11p) infection. First, we showed that CD46 molecules, which act as receptors for Ad11p, are expressed in AGMK cells. We then monitored Ad11p replication by measuring GFP expression as an indicator of viral transcription. We found that AGMK-derived cells were as capable as carcinoma cells at propagating full-length replication-competent Ad11p (RCAd11p) DNA. Of the AGMK cell lines tested, Vero cells had the greatest capacity for adenovirus production. Thus, AGMK cells can be used to evaluate RCAd11p-mediated gene delivery, and Vero cells can be used for the production of RCAd11pGFP vectors at relatively high yields. - Highlights: • Africa green monkey cell lines were monitored for human adenovirus 11p GFP vector infection. • Human CD46 molecules were detectable in these monkey cell lines. • Adenovirus 11p GFP vector can be propagated in Vero cells increases the safety of Ad11p-based vectors for clinical trials. • To use Vero cells for preparation of Ad11p vector avoids the potential inclusion of oncogenes from tumor cells.

  3. CaCl2-heat shock preparation of competent cells of three Pseudomonas strains and related transformation conditions%3种假单胞菌CaCl2法感受态细胞制备及转化条件

    Institute of Scientific and Technical Information of China (English)

    赵峰; 张颖; 李慧; 史荣久; 韩斯琴

    2013-01-01

    假单胞菌因其生境和代谢类型的多样性,在污染环境修复、生物转化、生物防治等领域具有广阔的应用潜力;外源基因的导入是假单胞菌遗传改造的重要环节,而感受态细胞的制备和转化方法的建立是导入外源基因的重要方法学基础.本研究以从石油污染土壤中分离筛选的假单胞菌属的3个菌株Pseudomonas putida TS11、P.stutzeri DNB、P.mendocina JJ12为对象,通过3因素4水平正交实验设计,研究了不同CaCl2浓度、热激时间及复苏时间对不同假单胞菌感受态细胞制备及转化效率的影响.结果表明:CaCl2浓度是影响假单胞菌转化效率的最主要因素(P<0.05),且在制备感受态细胞之前用无菌蒸馏水多次洗涤菌体细胞,转化率明显提高.3种假单胞菌的CaCl2转化优化条件分别为:100 mmol·L-1 CaCl2,热激3 min,复苏1.5 h;50 mmol·L-1 CaCl2,热激6 min,复苏1.5 h;75 mmol·L-1 CaCl2,热激4.5 min,复苏0.5h.在上述转化条件下,3种假单胞菌的外源质粒转化效率均达到105个转化子·μg-1DNA水平.%Pseudomonas, due to its diversity in habitat and metabolic type, makes it have broad prospects applying in bioremediation, byconversion, and biocontrol, while the introduction of exogenous gene is the key link to genetically modified Pseudomonas.The preparation and transformation of competent cells are the important methodological basis of the introduction of exogenous gene.In this paper, three Pseudomonas strains (P.putida TS11, P.stutzeri DNB, and P.mendocina JJ12) isolated from a petroleum-contaminated soil were taken as the recipient strains, and a three-factor and four-level orthogonal experiment was conducted to investigate the effects of CaCl2 concentration, heat shock duration, and recovery duration on the preparation and transformation efficiency of the strains competent cells.The results showed that CaCl2 concentration was the most important factor affecting the transformation

  4. Effectiveness of Simulation Preparation on Novice Nurses' Competence and Self-Efficacy in a Multiple-Patient Simulation.

    Science.gov (United States)

    Franklin, Ashley E; Gubrud-Howe, Paula; Sideras, Stephanie; Lee, Christopher S

    2015-01-01

    This study examined the influence of three simulation preparation methods (expert modeling/intervention, voice-over PowerPoint/active control, and reading assignments/passive control) on improving novice nurses' competence and self-efficacy for providing care to multiple simulated patients. Both competence and self-efficacy were measured at baseline and following a five-week intervention. Twenty senior pre-licensure nursing students participated in the trial. One-way ANOVAs and parametric/nonparametric correlations were used. Voice-over PowerPoint and expert modeling resulted in greater improvements in self-efficacy compared with traditional reading assignments as simulation preparation. However, there was no relationship between change in competence and self-efficacy over time.

  5. Using Interactive Online Role-Playing Simulations to Develop Global Competency and to Prepare Engineering Students for a Globalised World

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    May, Dominik; Wold, Kari; Moore, Stephanie

    2015-01-01

    The world is changing significantly, and it is becoming increasingly globalised. This means that countries, businesses, and professionals must think and act globally to be successful. Many individuals, however, are not prepared with the global competency skills needed to communicate and perform effectively in a globalised system. To address this…

  6. The Preparation of Pre-Service Student Teachers' Competence to Work in Schools

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    Tang, Sylvia Y. F.; Cheng, May M. H.; Wong, Angel K. Y.

    2016-01-01

    Competence to work in schools is an important dimension of professional competence, although it is often a neglected dimension of teacher development. This article reports a qualitative study that examined student teachers' learning experiences in initial teacher education (ITE) in relation to competence to work in schools. In-depth interviews…

  7. Pneumococcal Competence Coordination Relies on a Cell-Contact Sensing Mechanism.

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    Marc Prudhomme

    2016-06-01

    Full Text Available Bacteria have evolved various inducible genetic programs to face many types of stress that challenge their growth and survival. Competence is one such program. It enables genetic transformation, a major horizontal gene transfer process. Competence development in liquid cultures of Streptococcus pneumoniae is synchronized within the whole cell population. This collective behavior is known to depend on an exported signaling Competence Stimulating Peptide (CSP, whose action generates a positive feedback loop. However, it is unclear how this CSP-dependent population switch is coordinated. By monitoring spontaneous competence development in real time during growth of four distinct pneumococcal lineages, we have found that competence shift in the population relies on a self-activated cell fraction that arises via a growth time-dependent mechanism. We demonstrate that CSP remains bound to cells during this event, and conclude that the rate of competence development corresponds to the propagation of competence by contact between activated and quiescent cells. We validated this two-step cell-contact sensing mechanism by measuring competence development during co-cultivation of strains with altered capacity to produce or respond to CSP. Finally, we found that the membrane protein ComD retains the CSP, limiting its free diffusion in the medium. We propose that competence initiator cells originate stochastically in response to stress, to form a distinct subpopulation that then transmits the CSP by cell-cell contact.

  8. TO THE PROBLEM OF FORMATION OF PEDAGOGICAL COMPETENCE IN PROFESSIONAL PREPARATION OF MATHEMATIANS

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    S. P. GRUSHEVSKIY

    2016-01-01

    Full Text Available This article presents the approach to formation of pedagogical competences of mathematicians in the course of vocational training on the basis of realization of through trajectories of professional and pedagogical preparation (the bachelor + the master in conditions of a two-level system of higher education. In accordance with the Federal State Educational Standards of higher education bachelor of mathematics in directions 01.03.01 "Mathematics", 02.03.01 "Mathematics and Computer Science" pedagogical practice is highlighted as one of the professional activities of graduates who have mastered the undergraduate program. That university graduates always form the basis of the body of teachers of mathematics and computer science both a secondary and higher education institutions. However, due to problematic features of the educational standards of bachelor of mathematics training emphasis shifts toward a general theoretical pedagogical training and towards the formation of the basic components of vocational training. Lack of training time does not allow graduates to get the necessary experience with pedagogical knowledge, so psychological and pedagogical unit does not complete with obtaining of the teacher qualification. In order to solve the existing problem, we propose the formation of professional pedagogical competence in modern mathematical and pedagogical master's programs, completing vocational teacher training of mathematicians. This approach provides a pass-through trajectory of vocational teacher training scheme Bachelor + Master in a two-level structure of higher education, the effectiveness of which is confirmed by experience in implementing mathematical and pedagogical master's programs "Information Technology in Education" and "Teaching Mathematics and Computer Science" at the Faculty of Mathematics and Computer Science of Kuban State University. The article reveals the basic structural components of

  9. Crosswalking public health and health education competencies: implications for professional preparation and practice.

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    Woodhouse, Lynn D; Auld, M Elaine; Miner, Kathleen; Alley, Kelly Bishop; Lysoby, Linda; Livingood, William C

    2010-01-01

    This article highlights similarities and differences between the public health competencies recently developed by the Association of Schools of Public Health (ASPH) and one public health specialty, health education (HE), which has used competencies in its quality assurance systems for more than 20 years. Based on a crosswalk methodology developed for this analysis, some 50 percent to 61 percent of the HE and ASPH competencies had similarities of varying degrees; 18 percent were deemed matches due to sameness in skill or content. Most similarities were found between the ASPH social and behavioral sciences competencies and the HE competencies. Significant domains of "no match" were found between the HE and ASPH competencies in the areas of Systems Thinking, Leadership, and Public Health Biology. The study results have implications for academic programs related to curricula review and revision, continuing education providers who are developing training agendas for the workforce, employers anticipating competencies in new job hires, and prospective students and practitioners who are considering a form of certification. Qualitative insights from the study related to professional culture, purpose, age, and consistency of the scope or depth of the two competency sets, as well as the crosswalk methodology itself, may be useful to those comparing other competency sets.

  10. FUTURE TEACHER PROFESSIONAL PREPARATION IN COMPETENCE-VALUED DIMENSION: PHILOSOPHY-CULTURE ASPECT

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    Yurii V. Pelekh

    2010-08-01

    Full Text Available The article deals with some aspects of actualization of “competence” and “competent approach” notions in modern pedagogical science. The attempt of the definition of conceptual axiological statements in the competence theory structure on the basis of philosophical culturelogical analysis has been made.

  11. [Preparation of cytoplasts from HL-60 cells].

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    Wang, Lili; Yu, Huangfei; Fang, Ning; Chen, Daixiong

    2013-06-01

    This experimental research was aimed to establish an optimum system of enucleation, purification and identification for preparing the cytoplasts of suspension culture cells in order to undertake cell recombination. Human leukemia HL-60 cells in suspension culture were purified by 42% Percoll density gradient centrifugation and low-speed centrifugation at 1 500r/min, respectively. The purified HL-60 cells were treated with cytochalasin B (CB) alone or combined with colcchicine and enucleated by isopycnic gradient centrifugation on 50% Percoll at 25 degrees C and 34 degrees C, respectively. Cytoplasts made from HL-60 cells were purified through gradient centrifugation by 37%, 38% and 40% Percoll, respectively. The final cytoplasts were identified by Wright-Giemsa staining and 4,6-diamidino-2-phenylidole dihydrochloride (DAPI)/5, 6-carboxyflu-orescein diacetate succinimidyl ester (CFSE) double-staining. The phenotype and mitochondrial membrane potential of HL-60 cytoplasts were analyzed by flow cytometry. The results indicated that the enucleation ratio of HL-60 cells induced by CB combined with colcchicine was up to 91. 98% +/-4. 29%, which was significantly higher than that in CB alone group (74. 95% +/- 3. 02%)(PHL-60 cytoplasts had no significant change, and the activity of the cytoplasts was above 80% within 12h. It is concluded that enucleation throuth density gradient centrifugation on 50% Percoll mediated by CB combined with colcchicine, 38%Percoll of purification followed by DAPI/CFSE double labeling and MMP detection is an optimum scheme for preparation and identification of cytoplast from suspension culture cells.

  12. Preparing for the data revolution: identifying minimum health information competencies among the health workforce.

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    Whittaker, Maxine; Hodge, Nicola; Mares, Renata E; Rodney, Anna

    2015-04-01

    Health information is required for a variety of purposes at all levels of a health system, and a workforce skilled in collecting, analysing, presenting, and disseminating such information is essential to fulfil these demands. While it is established that low- and middle-income countries (LMICs) are facing shortages in human resources for health (HRH), there has been little systematic attention focussed on non-clinical competencies. In response, we developed a framework that defines the minimum health information competencies required by health workers at various levels of a health system. Using the Delphi method, we consulted with leading global health information system (HIS) experts. An initial list of competencies and draft framework were developed based on results of a systematic literature review. During the second half of 2012, we sampled 38 experts with broad-based HIS knowledge and extensive development experience. Two rounds of consultation were carried out with the same group to establish validity of the framework and gain feedback on the draft competencies. Responses from consultations were analysed using Qualtrics® software and content analysis. In round one, 17 experts agreed to participate in the consultation and 11 (65%) completed the survey. In the second round, 11 experts agreed to participate and eight (73%) completed the survey. Overall, respondents agreed that there is a need for all health workers to have basic HIS competencies and that the concept of a minimum HIS competency framework is valid. Consensus was reached around the inclusion of 68 competencies across four levels of a health system. This consultation is one of the first to identify the HIS competencies required among general health workers, as opposed to specialist HIS roles. It is also one of the first attempts to develop a framework on minimum HIS competencies needed in LMICs, highlighting the skills needed at each level of the system, and identifying potential gaps in current

  13. Using interactive online role-playing simulations to develop global competency and to prepare engineering students for a globalised world

    Science.gov (United States)

    May, Dominik; Wold, Kari; Moore, Stephanie

    2015-09-01

    The world is changing significantly, and it is becoming increasingly globalised. This means that countries, businesses, and professionals must think and act globally to be successful. Many individuals, however, are not prepared with the global competency skills needed to communicate and perform effectively in a globalised system. To address this need, higher education institutions are looking for ways to instil these skills in their students. This paper explains one promising approach using current learning principles: transnational interactive online environments in engineering education. In 2011, the TU Dortmund and the University of Virginia initiated a collaboration in which engineering students from both universities took part in one online synchronous course and worked together on global topics. This paper describes how the course was designed and discusses specific research results regarding how interactive online role-playing simulations support students in gaining the global competency skills required to actively participate in today's international workforce.

  14. Special Education Teacher Transition-Related Competencies and Preparation in Saudi Arabia

    Science.gov (United States)

    Alnahdi, Ghaleb

    2014-01-01

    Preparing special education teachers to engage in transition services is a critical part of their preparation. This study examined how special education teachers perceive their preparation for transition services in Riyadh, Saudi Arabia. A total of 350 teachers participated in this study. Data were analyzed using a two-way ANOVA. The findings…

  15. Efficient Induction and Isolation of Human Primordial Germ Cell-Like Cells from Competent Human Pluripotent Stem Cells.

    Science.gov (United States)

    Irie, Naoko; Surani, M Azim

    2017-01-01

    We recently reported a robust and defined culture system for the specification of human primordial germ cell-like cells (hPGCLCs) from human pluripotent stem cells (hPSCs), both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in vitro (Irie et al. Cell 160: 253-268, 2015). Similar attempts previously produced hPGCLCs from hPSCs at a very low efficiency, and the resulting cells were not fully characterized. A key step, which facilitated efficient hPGCLC specification from hPSCs, was the induction of a "competent" state for PGC fate via the medium containing a cocktail of four inhibitors. The competency of hPSCs can be maintained indefinitely and interchangeably with the conventional/low-competent hPSCs. Specification of hPGCLC occurs following sequential expression of key germ cell fate regulators, notably SOX17 and BLIMP1, as well as initiation of epigenetic resetting over 5 days. The hPGCLCs can be isolated using specific cell surface markers without the need for generating germ cell-specific reporter hPSC lines. This powerful method for the induction and isolation of hPGCLCs can be applied to both hESCs and iPSCs, which can be used for advances in human germ line biology.

  16. How Two Small Pharmacy Schools’ Competency Standards Compare with an International Competency Framework and How Well These Schools Prepare Students for International Placements

    Directory of Open Access Journals (Sweden)

    John Hawboldt

    2017-03-01

    Full Text Available International standards of pharmacy curricula are necessary to ensure student readiness for international placements. This paper explores whether curricula from two pharmacy programs, in Australia and Canada, are congruent with international standards and if students feel prepared for international placements. Nationally prescribed educational standards for the two schools were compared to each other and then against the International Pharmaceutical Federation (FIP Global Competency Framework. Written student reflections complemented this analysis. Mapping results suggested substantial agreement between the FIP framework and Australia and Canada, with two gaps being identified. Moreover, the students felt their programs prepared them for their international placements. Despite differences in countries, pharmacy programs, and health-systems all students acclimatized to their new practice sites. Implications are that if pharmacy programs align well with FIP, pharmacists should be able to integrate and practise in other jurisdictions that also align with the FIP. This has implications for the mobility of pharmacy practitioners to countries not of their origin of training.

  17. A Comparison of Three Teacher Preparation Programs in Terms of Their Impact on Teacher Competencies

    Science.gov (United States)

    Yao, Yuankun; Williams, Wayne

    2010-01-01

    This study compared the effectiveness of three teacher education programs in terms of their impact on teacher competencies. The programs include a traditional 4-year teacher certification program, an alternative certification-based master of arts in teaching program, and an alternative certification program without the master degree option. The…

  18. Allied Health Care Employees' Workplace Skills and Competencies: Are They Prepared?

    Science.gov (United States)

    McClain, Clifford R.; McClain, Mildred A.

    2007-01-01

    The purpose of this study was to determine the extent to which allied health care providers considered the Secretary's Commission on Achieving Necessary Skills (SCANS, 1991) and competencies as those that are necessary for entry level employment in the allied health care industry. The extent that allied health care supervisors and managers…

  19. Teacher Preparation for Movement Education: Increasing Pre-Service Teachers' Competence for Working with Young Children

    Science.gov (United States)

    Sevimli-Celik, Serap; Johnson, James E.

    2016-01-01

    This study explores pre-service teachers' perceptions of movement education, the benefits they perceive from participating in a 12-week movement education module in a course on play, and the module's effects on their confidence and competence in regard to incorporating movement into a curriculum. Findings suggest that the pre-service teachers…

  20. A Study Examining the Dimensionality of Core Competencies Measure in Teacher Preparation Programs: Challenges and Lessons

    Science.gov (United States)

    Kizil, Ruhan Circi; Briggs, Derek; Seidel, Kent; Green, Kathy

    2014-01-01

    The evidence that teacher preparation programs have an impact on teacher quality is often limited. Progress in research on this topic will remain rather limited in its influence on practice until more proximal measures of teacher education outcomes can be established. The dearth of variables to measure the impact of teacher preparation programs on…

  1. 76 FR 59329 - User Fees Relating to the Registered Tax Return Preparer Competency Examination and...

    Science.gov (United States)

    2011-09-26

    ... retirement plan agent, or enrolled actuary who signs the tax return or claim for refund prepared by the..., certified public accountants, enrolled agents, enrolled retirement plan agents, and enrolled actuaries...

  2. Stereological quantification of immune-competent cells in baseline biopsy specimens from achilles tendons

    DEFF Research Database (Denmark)

    Kragsnaes, Maja Skov; Fredberg, Ulrich; Stribolt, Katrine

    2014-01-01

    immunohistochemically by quantifying the presence of macrophages (CD68-PGM1(+), CD68-KP1(+)), hemosiderophages (Perls blue), T lymphocytes (CD2(+), CD3(+), CD4(+), CD7(+), CD8(+)), B lymphocytes (CD20(+)), natural killer cells (CD56(+)), mast cells (NaSDCl(+)), Schwann cells (S100(+)), and endothelial cells (CD34......(+)) using a stereological technique. A follow-up examination was conducted more than 4 years (range, 4-9 years) after the biopsy procedure to evaluate the long-term presence of Achilles tendon symptoms. RESULTS: Macrophages, T lymphocytes, mast cells, and natural killer cells were observed in the majority......BACKGROUND: Limited data exist on the presence and function of immune-competent cells in chronic tendinopathic tendons and their potential role in inflammation and tissue healing as well as in predicting long-term outcome. PURPOSE: To quantify subtypes of immune-competent cells in biopsy specimens...

  3. Preparing Teachers for Technology Integration: Programs, Competencies, and Factors from the Literature

    Science.gov (United States)

    Oliver, Kevin; Townsend, Latricia

    2013-01-01

    This article presents a review of recent literature about preparing teachers for technology integration. The review found six types of training programs are commonly implemented: pre-service training, long-term courses, short-term workshops and institutes, coaching/mentoring, learning communities, and product/assessment approaches. The review…

  4. Preparing Community College Leaders: The AACC Core Competencies for Effective Leadership & Doctoral Education

    Science.gov (United States)

    McNair, Delores E.

    2010-01-01

    Community colleges in the United States face shortages of leaders prepared to assume administrative positions in the 21st century. To respond to this shortage, graduate programs are emerging with a specific emphasis on community college leadership; other graduate programs offer broader curricula focused on educational leadership, policy, or higher…

  5. Preparing Community College Leaders: The AACC Core Competencies for Effective Leadership & Doctoral Education

    Science.gov (United States)

    McNair, Delores E.

    2010-01-01

    Community colleges in the United States face shortages of leaders prepared to assume administrative positions in the 21st century. To respond to this shortage, graduate programs are emerging with a specific emphasis on community college leadership; other graduate programs offer broader curricula focused on educational leadership, policy, or higher…

  6. Preparing Culturally and Linguistically Competent Teachers for English as an International Language Education

    Science.gov (United States)

    Li, Guofang

    2017-01-01

    Despite recent scholarly advancements in teaching English as an international language (EIL), its implementation in TESOL classrooms has been challenging and limited. Because English teachers play a significant role in EIL implementation in their daily lessons, preparing EIL-oriented teachers becomes critical. This article outlines major…

  7. Resistance to Racial/Ethnic Dialog in Graduate Preparation Programs: Implications for Developing Multicultural Competence

    Science.gov (United States)

    Kelly, Bridget Turner; Gayles, Joy Gaston

    2010-01-01

    This study sought to understand how individuals experienced multicultural courses in graduate preparation programs. The researchers conducted focus groups with 37 current and former graduate students in student affairs. Participants reported resistance to multicultural dialog, both in their direct experiences and through their perceptions of…

  8. Preparing a 21st century workforce: is it time to consider clinically based, competency-based training of health practitioners?

    Science.gov (United States)

    Nancarrow, Susan A; Moran, Anna M; Graham, Iain

    2014-02-01

    Health workforce training in the 21st century is still based largely on 20th century healthcare paradigms that emphasise professionalisation at the expense of patient-focussed care. This is illustrated by the paradox of increased training times for health workers that have corresponded with workforce shortages, the limited career options and pathways for paraprofessional workers, and inefficient clinical training models that detract from, rather than add to, service capacity. We propose instead that a 21st century health workforce training model should be: situated in the clinical setting and supported by outsourced university training (not the other way around); based on the achievement of specific milestones rather than being time-defined; and incorporate para-professional career pathways that allow trainees to 'step-off' with a useable qualification following the achievement of specific competencies. Such a model could be facilitated by existing technology and clinical training infrastructure, with enormous potential for economies of scale in the provision of formal training. The benefits of a clinically based, competency-based model include an increase in clinical service capacity, and clinical training resources become a resource for the delivery of healthcare, not just education. Existing training models are unsustainable, and are not preparing a workforce with the flexibility the 21st century demands.

  9. Germ Cells are Made Semiotically Competent During Evolution

    DEFF Research Database (Denmark)

    Giorgi, Franco; Bruni, Luis Emilio

    2016-01-01

    Germ cells are cross-roads of development and evolution. They define the origin of every new generation and, at the same time, represent the biological end-product of any mature organism. Germ cells are endowed with the following capacities: (1) to store a self-descriptive program, (2) to accumul......Germ cells are cross-roads of development and evolution. They define the origin of every new generation and, at the same time, represent the biological end-product of any mature organism. Germ cells are endowed with the following capacities: (1) to store a self-descriptive program, (2......-ended semiotic relationship explored and gradually defined during evolution by the context-dependency of specific cell-to-cell interactions. In this way, any structural and functional novelty that has emerged in the course of germ cell interactions may be interpreted as an exaptation fixed in the species genome...

  10. The big Cs - contemplating on the competency and credibility of master's-prepared early career nurses.

    Science.gov (United States)

    Liu, Linda

    2014-09-01

    The landscape in master's-prepared nursing education has vastly changed. Programs are constantly expanding and evolving to respond to changing trends and needs in healthcare. Simultaneously, the demographic profile of nurses returning to graduate studies has immeasurably diversified. While it can be argued that developing advanced practice nurses (APNs) at an earlier time point may lengthen their leadership careers such that they build stronger portfolios to effect more substantive healthcare changes, it is also worth questioning whether limited clinical experience in our new generation of nurse leaders will ultimately widen our proverbial knowledge-practice gap.

  11. Maf acts downstream of ComGA to arrest cell division in competent cells of B. subtilis.

    Science.gov (United States)

    Briley, Kenneth; Prepiak, Peter; Dias, Miguel J; Hahn, Jeanette; Dubnau, David

    2011-07-01

    Transformable (competent) cells of Bacillus subtilis are blocked in cell division because the traffic ATPase ComGA prevents the formation of FtsZ rings. Although ComGA-deficient cells elongate and form FtsZ rings, cell division remains blocked at a later stage and the cells become mildly filamented. Here we show that the highly conserved protein Maf is synthesized predominantly in competent cells under the direct control of the transcription factor ComK and is solely responsible for the later block in cell division. In vivo and in vitro data show that Maf binds to both ComGA and DivIVA. A point mutation in maf that interferes with Maf binding to DivIVA also interferes with the ability of Maf to inhibit cell division. Based on these findings, we propose that Maf and ComGA mediate mechanisms for the inhibition of cell division in competent cells with Maf acting downstream of ComGA. We further suggest that Maf must interact with DivIVA to inhibit cell division. © 2011 Blackwell Publishing Ltd.

  12. Isolation and catalytic competence of different animal liver microsomal fractions prepared by calcium-aggregation method

    Directory of Open Access Journals (Sweden)

    Walawalkar Pradnya

    2006-01-01

    Full Text Available Rat, mouse, rabbit, and guinea pig liver microsomes were prepared, using the calcium-aggregation method. The mean specific spectral cytochrome P450 content obtained for rat, mouse, rabbit, and guinea pig liver microsomes were 0.547, 0.394, 0.677, and 0.378 nmol cytochrome P450/mg protein, respectively. All the microsomal samples showed the ability to biotransform p-nitrophenol to p-nitrocatechol. The mean rate of formation of p-nitrocatechol by rat, mouse, rabbit, and guinea pig liver microsomes was 0.272, 0.247, 0.497 and 0.424 nmole/min/nmole cytochrome 450, respectively. This method circumvents the need of an ultracentrifuge for isolation of microsomes, by the conventional differential centrifugation method.

  13. Electron microscopy of single-stranded structures in the DNA of competent Haemophilus influenzae cells

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Kupfer, D.M.

    1987-02-01

    Chromosomal DNAs from exponential-phase and competent cells of Haemophilus influenzae were examined by electron microscopy to determine whether the chromosome undergoes structural changes during competence development. Single-stranded gaps and single-stranded tails formed in chromosomal DNA during competence development. The generation of gaps was dependent on the rec-2 function. Since the rec-2 mutant is defective in the translocation of donor DNA, it was inferred that the gaps were involved in the translocation step of transformation. The generation of single-stranded tails was independent of the rec-1 and rec-2 genes. Therefore, these structures were assumed to play no direct role in the interaction of donor and recipient DNAs during transformation. Gaps were preferentially associated with a readily denaturable, possible A + T-rich fraction of the genome. This finding raised the possibility that hot spots for transformation might be associated with A + T-rich DNA.

  14. Competency development in antibody production in cancer cell biology

    Energy Technology Data Exchange (ETDEWEB)

    Park, M.S.

    1998-12-01

    This is the final report of a three-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). The main objective of this project was to develop a rapid recombinant antibody production technology. To achieve the objective, the authors employed (1) production of recombinant antigens that are important for cell cycle regulation and DNA repair, (2) immunization and specific selection of antibody-producing lymphocytes using the flow cytometry and magnetic bead capturing procedure, (3) construction of single chain antibody library, (4) development of recombinant vectors that target, express, and regulate the expression of intracellular antibodies, and (5) specific inhibition of tumor cell growth in tissue culture. The authors have accomplished (1) optimization of a selection procedure to isolate antigen-specific lymphocytes, (2) optimization of the construction of a single-chain antibody library, and (3) development of a new antibody expression vector for intracellular immunization. The future direction of this research is to continue to test the potential use of the intracellular immunization procedure as a tool to study functions of biological molecules and as an immuno-cancer therapy procedure to inhibit the growth of cancer cells.

  15. Meristematic competence is disrupted by microgravity, real or simulated, in seedlings and cultured cells of Arabidopsis

    Science.gov (United States)

    Medina, Francisco Javier; Herranz, Raul; Van Loon, ing.. Jack J. W. A.; Kiss, John; Valbuena, Miguel A.; Youssef, Khaled

    In actively proliferating plant cells, the rate of cell proliferation is strictly coordinated with cell growth, and this coordination is called “meristematic competence”. Cell proliferation consists of the adequate progression of the cell division cycle throughout specific regulatory checkpoints, and cell growth consists of reaching the critical size making possible cell division, based on the increase of biomass, essentially by means of protein synthesis. There are two cellular models in which these processes can be studied, namely the meristematic tissues of plants and seedlings and the in vitro suspension cell cultures. Meristems are essential for the determination of the developmental pattern of the plant, which is primarily based on the balance between proliferating (meristematic) and differentiated cells. Auxin is a fundamental phytohormone, responsible for the maintenance of meristematic competence and for the control of the rate of differentiation. We first studied the proliferating activity of root meristematic cells in the International Space Station (ISS) and in a random positioning machine (RPM), a ground-based device for simulated microgravity. The result in both experiments was the increase of mitotic activity (cell proliferation) and the depletion of ribosome synthesis (cell growth), that is, the disruption of meristematic competence. We found these effects associated with changes in the auxin levels and polar transport, which is related to the role of auxin as a mediator of the transduction of the gravitropic signal sensed in the root columella. We plan to advance in the investigation of mechanisms of the auxin control of meristematic competence in microgravity conditions in a new experiment, “Seedling Growth”, to be performed in the ISS. We will use mutants of the auxin transport pathway and we will also test the potential activating role of red light, known to be a cell proliferation and gene expression enhancer. The role played by

  16. Preparation, cryopreservation, and growth of cells prepared from the green turtle (Chelonia mydas)

    Science.gov (United States)

    Moore, M.K.; Work, T.M.; Balazs, G.H.; Docherty, D.E.

    1997-01-01

    Techniques are described for preparing, preserving, and growing cell cultures from 30 to 40-day old green turtle embryos (2.0-3.0 cm length) including cells derived from skeletal muscle, liver, heart, kidney, eye, lung, and brain. Acceptable growth of all cells occurred in all standard cell culture media tested, with optimum growth temperature near 30??C. These cell cultures will be used in the study of sea turtle viral diseases including fibropapillomatosis, which is currently epidemic in some green turtle populations.

  17. The effects of roscovitine on cumulus cell apoptosis and the developmental competence of domestic cat oocytes.

    Science.gov (United States)

    Sananmuang, T; Techakumphu, M; Tharasanit, T

    2010-01-15

    The developmental competence of cat oocytes matured in vitro is relatively poor when compared with that of in vivo oocytes. The study aimed to investigate the effect of roscovitine on the developmental competence of cat Felis catus oocytes matured in vitro. Cumulus-oocyte complexes (COCs) were classified as Grade I and II to III. Groups of COCs were cultured in 0, 12.5, 25, 50, 100, and 200 microM roscovitine for 24h and were either fixed to assess the stages of nuclear maturation (Experiment 1) or additionally matured in vitro for 24h before fixation (Experiment 2). In Experiment 3, cumulus cells from the COCs treated with roscovitine were examined for apoptosis. Experiment 4 examined the developmental competence of cat oocytes after roscovitine treatment and in vitro fertilization in terms of cleavage and morula and blastocyst formation rates. Roscovitine reversibly arrested cat oocytes at an immature stage in a dose-dependent manner. Roscovitine at 12.5 and 25 microM demonstrated less efficiency compared with that of other doses. However, higher doses of roscovitine induced cumulus cell apoptosis and resulted in a high number of degenerated oocytes after in vitro maturation. Roscovitine at 12.5 and 25 microM were therefore used to evaluate their effect on embryo development. Pretreatment with 12.5 and 25 microM roscovitine prior to in vitro maturation decreased the developmental competence of cat oocytes compared with that of non-roscovitine-treated controls. In conclusion, roscovitine reversibly maintained cat oocytes at the germinal vesicle stage without detrimental effect on nuclear maturation. However, it negatively affected cumulus cell viability and developmental competence.

  18. Preparation of Single Cells for Imaging Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Berman, E S; Fortson, S L; Kulp, K S; Checchi, K D; Wu, L; Felton, J S; Wu, K J

    2007-10-24

    Characterizing chemical changes within single cells is important for determining fundamental mechanisms of biological processes that will lead to new biological insights and improved disease understanding. Imaging biological systems with mass spectrometry (MS) has gained popularity in recent years as a method for creating precise chemical maps of biological samples. In order to obtain high-quality mass spectral images that provide relevant molecular information about individual cells, samples must be prepared so that salts and other cell-culture components are removed from the cell surface and the cell contents are rendered accessible to the desorption beam. We have designed a cellular preparation protocol for imaging MS that preserves the cellular contents for investigation and removes the majority of the interfering species from the extracellular matrix. Using this method, we obtain excellent imaging results and reproducibility in three diverse cell types: MCF7 human breast cancer cells, Madin-Darby canine kidney (MDCK) cells, and NIH/3T3 mouse fibroblasts. This preparation technique allows routine imaging MS analysis of cultured cells, allowing for any number of experiments aimed at furthering scientific understanding of molecular processes within individual cells.

  19. Dynamic clustering and dispersion of lipid rafts contribute to fusion competence of myogenic cells

    Energy Technology Data Exchange (ETDEWEB)

    Mukai, Atsushi [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 36-3 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Kurisaki, Tomohiro [Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507 (Japan); Sato, Satoshi B. [Research Center for Low Temperature and Material Sciences, Kyoto University, Yoshida-honmachi, Kyoto 606-8501 (Japan); Kobayashi, Toshihide [Lipid Biology Laboratory, Discovery Research Institute, RIKEN, Wako, Saitama 351-0198 (Japan); Kondoh, Gen [Laboratory of Animal Experiments for Regeneration, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507 (Japan); Hashimoto, Naohiro, E-mail: nao@nils.go.jp [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 36-3 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan)

    2009-10-15

    Recent research indicates that the leading edge of lamellipodia of myogenic cells (myoblasts and myotubes) contains presumptive fusion sites, yet the mechanisms that render the plasma membrane fusion-competent remain largely unknown. Here we show that dynamic clustering and dispersion of lipid rafts contribute to both cell adhesion and plasma membrane union during myogenic cell fusion. Adhesion-complex proteins including M-cadherin, {beta}-catenin, and p120-catenin accumulated at the leading edge of lamellipodia, which contains the presumptive fusion sites of the plasma membrane, in a lipid raft-dependent fashion prior to cell contact. In addition, disruption of lipid rafts by cholesterol depletion directly prevented the membrane union of myogenic cell fusion. Time-lapse recording showed that lipid rafts were laterally dispersed from the center of the lamellipodia prior to membrane fusion. Adhesion proteins that had accumulated at lipid rafts were also removed from the presumptive fusion sites when lipid rafts were laterally dispersed. The resultant lipid raft- and adhesion complex-free area at the leading edge fused with the opposing plasma membrane. These results demonstrate a key role for dynamic clustering/dispersion of lipid rafts in establishing fusion-competent sites of the myogenic cell membrane, providing a novel mechanistic insight into the regulation of myogenic cell fusion.

  20. Microvesicles preparation from mesenchymal stem cells.

    Science.gov (United States)

    Rad, Fariba; Pourfathollah, Ali Akbar; Yari, Fatemeh; Mohammadi, Saeed; Kheirandish, Maryam

    2016-01-01

    Extracellular vesicles are particles ranged from 30 nm to 5μm and subcategorized into three groups; exosomes, microvesicles and apoptotic bodies, each of which have different biological impact. Lack of a standard method for the detection and isolation of MVs has led to a challenging issue that is a worth considering. In this study, we isolated MVs from the conditioned medium of UC-MSCs by four different schemes of ultracentrifugation. We examined the efficacy of differential centrifugation ranging from 10,000×g to 60,000×g on UCMSCs- derived microvesicles yield and purity. The fractions were evaluated by Dynamic Light Scattering (DLS) method, total protein quantification and flow cytometry. UC-MSCs were spindle cells that adhered to plastic culture flasks. These cells expressed MSC markers such as CD44 and CD73, whereas were negative for hematopoietic markers CD45 and CD34. UC-MSCparticles were successfully isolated. Particles were heterogeneous vesicles of approximately 50 to 1250 nm in diameter that bear the surface-expressed molecules UC-MSCs such as; CD90, CD106, CD166 and CD44, and negative for CD34, CD63, and CD9. According to the results of DLS method, centrifugation at 10,000, 20,000, 40,000 and 60,000 ×g, all gave MVs of less than 1000 nm. It is of notion that only at the centrifugation rates of 40,000 and 60,000×g, particles of less than 100 nm in diameter were also obtained. The choice of exact speed greatly influences the purity of MVs and their yield. Our findings indicate that centrifugation at 20,000×g is appropriate for the purification of UC-MSC-MVs.

  1. Preparing Pre-Service Teachers to Integrate Technology: An Analysis of the Emphasis on Digital Competence in Teacher Education Curricula

    Science.gov (United States)

    Instefjord, Elen; Munthe, Elaine

    2016-01-01

    This article focuses on integration of digital competence in curriculum documents for teacher education in Norway. A model inspired by the work of Zhao, Pugh, Sheldon and Byers, as well as Krumsvik and Mishra and Koehler, has been developed as an analytical framework. Teachers' digital competence is here understood as comprising three knowledge…

  2. Determining the directions of increasing the innovative potential of the region by developing innovative technologies and competences when preparing and implementing large investment projects

    Directory of Open Access Journals (Sweden)

    Timur V. Kramin

    2015-12-01

    Full Text Available Objective to determine the directions of increasing the innovative potential of a region through the development of innovative technologies and competences in the process of preparation and implementation of large investment projects in the Republic of Tatarstan. Methods methodology of project management institutional approach. Results it is proved that the main largescale directions of innovative potential development in the Republic of Tatarstan as a result of preparation and implementing of Universiade 2013 in Kazan are knowledge management information technologies risk management. It is shown that in the framework of the considered innovative areas a complete system was formed of competences of employees and managers in the fields of education trade hospitality and service. Scientific novelty the key directions were defined of increasing the innovative potential of a region through the development of innovative technologies and competences in the process of preparation and implementation of large investment projects by the example of the World Summer Student Games in Kazan in 2013. Practical significance on the basis of specific examples the authors illustrate the practiceoriented mechanism of innovative potential development of a region as a result of implementation of large investment projects. nbsp

  3. REST controls self-renewal and tumorigenic competence of human glioblastoma cells.

    Science.gov (United States)

    Conti, Luciano; Crisafulli, Laura; Caldera, Valentina; Tortoreto, Monica; Brilli, Elisa; Conforti, Paola; Zunino, Franco; Magrassi, Lorenzo; Schiffer, Davide; Cattaneo, Elena

    2012-01-01

    The Repressor Element 1 Silencing Transcription factor (REST/NRSF) is a master repressor of neuronal programs in non-neuronal lineages shown to function as a central regulator of developmental programs and stem cell physiology. Aberrant REST function has been associated with a number of pathological conditions. In cancer biology, REST has been shown to play a tumor suppressor activity in epithelial cancers but an oncogenic role in brain childhood malignancies such as neuroblastoma and medulloblastoma. Here we examined REST expression in human glioblastoma multiforme (GBM) specimens and its role in GBM cells carrying self-renewal and tumorigenic competence. We found REST to be expressed in GBM specimens, its presence being particularly enriched in tumor cells in the perivascular compartment. Significantly, REST is highly expressed in self-renewing tumorigenic-competent GBM cells and its knock down strongly reduces their self-renewal in vitro and tumor-initiating capacity in vivo and affects levels of miR-124 and its downstream targets. These results indicate that REST contributes to GBM maintenance by affecting its self-renewing and tumorigenic cellular component and that, hence, a better understanding of these circuitries in these cells might lead to new exploitable therapeutic targets.

  4. Global competencies of regional stem cell research: bibliometrics for investigating and forecasting research trends.

    Science.gov (United States)

    Watatani, Kenji; Xie, Zhongquan; Nakatsuji, Norio; Sengoku, Shintaro

    2013-09-01

    We employed a bibliometric approach to examine regional stem cell research in the USA, the UK, Japan and China based on publications from 2007 to 2011 with a co-citation clustering analysis to identify region-specific clusters of global competencies. We observed that there are clear differences in the number and interdisciplinary spread of competencies across regions: the USA retains the largest capacity and capability for pursuing medical and pharmaceutical applications; China has shown substantial growth through fusion approaches with chemistry and material sciences; Japan has been pursuing basic biology and is currently seeking further growth; and the UK has shown considerable growth and quality with a focus on medical research and the widest interdisciplinary spread. Furthermore, we discuss policy implications from these results in terms of industrial and clinical applications. These findings provide a rational way of evaluating research policies and forecasting research trends.

  5. Replication-competent chimeric lenti-oncovirus with expanded host cell tropism.

    Science.gov (United States)

    Reiprich, S; Gundlach, B R; Fleckenstein, B; Uberla, K

    1997-04-01

    Baboon bone marrow was grafted into human immunodeficiency virus type 1-infected patients in the course of recent trials for AIDS treatment. Since the baboon genome harbors multiple copies of an endogenous oncovirus, chimeric lenti-oncoviruses could emerge in the xenotransplant recipient. To analyze the potential replication competence of hybrid viruses between different genera of retroviruses, we replaced most of the env gene of simian immunodeficiency virus with the env gene of an amphotropic murine leukemia virus. The hybrid virus could be propagated in human T-cell lines, in peripheral blood mononuclear cells of rhesus macaques, and in CD4- B-cell lines. Because of the expanded cell tropism, the hybrid virus might have a selective advantage in comparison to parental viruses. Therefore, emerging chimeric viruses may be considered a serious risk of xenotransplantation. A note of caution is also suggested for the use of pseudotyped lentiviral vectors for human gene therapy.

  6. Gluco-incretins regulate beta-cell glucose competence by epigenetic silencing of Fxyd3 expression.

    Directory of Open Access Journals (Sweden)

    David Vallois

    Full Text Available Gluco-incretin hormones increase the glucose competence of pancreatic beta-cells by incompletely characterized mechanisms.We searched for genes that were differentially expressed in islets from control and Glp1r-/-; Gipr-/- (dKO mice, which show reduced glucose competence. Overexpression and knockdown studies; insulin secretion analysis; analysis of gene expression in islets from control and diabetic mice and humans as well as gene methylation and transcriptional analysis were performed.Fxyd3 was the most up-regulated gene in glucose incompetent islets from dKO mice. When overexpressed in beta-cells Fxyd3 reduced glucose-induced insulin secretion by acting downstream of plasma membrane depolarization and Ca++ influx. Fxyd3 expression was not acutely regulated by cAMP raising agents in either control or dKO adult islets. Instead, expression of Fxyd3 was controlled by methylation of CpGs present in its proximal promoter region. Increased promoter methylation reduced Fxyd3 transcription as assessed by lower abundance of H3K4me3 at the transcriptional start site and in transcription reporter assays. This epigenetic imprinting was initiated perinatally and fully established in adult islets. Glucose incompetent islets from diabetic mice and humans showed increased expression of Fxyd3 and reduced promoter methylation.Because gluco-incretin secretion depends on feeding the epigenetic regulation of Fxyd3 expression may link nutrition in early life to establishment of adult beta-cell glucose competence; this epigenetic control is, however, lost in diabetes possibly as a result of gluco-incretin resistance and/or de-differentiation of beta-cells that are associated with the development of type 2 diabetes.

  7. Oocyte induction of EGF responsiveness in somatic cells is associated with the acquisition of porcine oocyte developmental competence.

    Science.gov (United States)

    Ritter, Lesley J; Sugimura, Satoshi; Gilchrist, Robert B

    2015-06-01

    Oocytes progressively acquire the competence to support embryo development as oogenesis proceeds with ovarian folliculogenesis. The objectives of this study were to investigate oocyte-secreted factor (OSF) participation in the development of somatic cell epidermal growth factor (EGF) responsiveness associated with oocyte developmental competence. A well-established porcine model was employed using oocytes from small (4 mm) antral follicles, representing low vs moderate developmental competence, respectively. Cumulus-oocyte complexes (COCs) were treated in vitro with inducers of oocyte maturation, and cumulus cell functions and oocyte developmental competence were assessed. COCs from small follicles responded to FSH but, unlike COCs from larger follicles, were incapable of responding to EGF family growth factors known to mediate oocyte maturation in vivo, exhibiting perturbed cumulus expansion and expression of associated transcripts (HAS2 and TNFAIP6). Low and moderate competence COCs expressed equivalent levels of EGF receptor (EGFR) mRNA; however, the former had less total EGFR protein leading to failed activation of phospho-EGFR and phospho-ERK1/2, despite equivalent total ERK1/2 protein levels. Native OSFs from moderate, but not from low, competence oocytes established EGF responsiveness in low competence COCs. Four candidate recombinant OSFs failed to mimic the actions of native OSFs in regulating cumulus expansion. Treatment with OSFs and EGF enhanced oocyte competence but only of the low competence COCs. These data suggest that developmental acquisition by the oocyte of capacity to regulate EGF responsiveness in the oocyte's somatic cells is a major milestone in the oocyte's developmental program and contributes to coordinated oocyte and somatic cell development.

  8. Jak Inhibitors Modulate Production of Replication Competent Zika Virus in Human Hofbauer, Trophoblasts, and Neuroblastoma cells

    Directory of Open Access Journals (Sweden)

    Christina Gavegnano

    2017-05-01

    Full Text Available Zika Virus (ZIKV is a Flavivirus that has been implicated in brain deformations, birth defects, and microcephaly of unborn fetuses and associated with Guillain-Barre syndrome.  Mechanisms responsible for transmission of ZIKV across the placenta to the fetus are incompletely understood.  Herein, we define key events modulating infection in clinically relevant cells, including primary placental macrophages (human hofbauer cells; HC, trophoblasts, and neuroblastoma cells. Consistent with previous findings, HC and trophoblasts are permissive to ZIKV infection. Decrease of interferon signaling by Jak 1/2 inhibition (via ruxolitinib significantly increased ZIKV replicationin HC, trophoblasts, and neuroblasts. Enhanced ZIKV production in ruxolitinib treated HC was associated with increased expression of HLA-DR and DC-SIGN. Nucleoside analogs blocked ruxolitinib-mediated production of extracellular virus. Although low-level ZIKV infection occurred in untreated HC and trophoblasts, the produced virus was incapable of infecting naïve Vero cells.  These deficient virions from untreated HC present “thin-coats” suggesting immature virion structure. Blocking Jak 1/2 signaling (with ruxolitinib restored replication competence as virions produced under these conditions confer CPE in naïve Vero cells.  These data demonstrate that Jak-STAT signaling directly impacts the ability of primary placental cells to produce replication competent virus and is a key gatekeeper in production of mature virions in clinically relevant cells including HC and trophoblasts. Design of targeted agents to prevent ZIKV replication in the placenta should consider Jak 1/2 signaling and the impact of its block on ZIKV infection and subsequent transmission to the fetus.

  9. Temozolomide competes for P-glycoprotein and contributes to chemoresistance in glioblastoma cells.

    Science.gov (United States)

    Munoz, Jessian L; Walker, Nykia D; Scotto, Kathleen W; Rameshwar, Pranela

    2015-10-10

    Chemotherapeutic resistance can occur by P-glycoprotein (P-gp), a 12-transmembrane ATP-dependent drug efflux pump. Glioblastoma (GBM) has poor survival rate and uniformly acquired chemoresistance to its frontline agent, Temozolomide (TMZ). Despite much effort, overcoming TMZ resistance remains a challenge. We reported on autonomous induction of TMZ resistance by increased transcription MDR1, the gene for P-gp. This study investigated how P-gp and TMZ interact to gain resistance. Using an experimental model of Adriamycin-resistant DC3F cells (DC3F/Adx), we showed that increased P-gp caused TMZ resistance. Increasing concentrations of TMZ competed with Calcein for P-gp, resulting in reduced efflux in the DC3F/Adx cells. Three different inhibitors of P-gp reversed the resistance to TMZ in two different GBM cell lines, by increasing active Caspase 3. Molecular modeling predicted the binding sites to be the intracellular region of P-gp and also identified specific amino acids and kinetics of energy for the efflux of TMZ. Taken together, we confirmed P-gp targeting of TMZ, a crucial regulator of TMZ resistance in GBM. This study provides insights on the effectiveness by which TMZ competes with other P-gp substrates, thereby opening the door for combined targeted therapies.

  10. Keratocyte fragments and cells utilize competing pathways to move in opposite directions in an electric field.

    Science.gov (United States)

    Sun, Yaohui; Do, Hao; Gao, Jing; Zhao, Ren; Zhao, Min; Mogilner, Alex

    2013-04-08

    Sensing of an electric field (EF) by cells-galvanotaxis-is important in wound healing [1], development [2], cell division, nerve growth, and angiogenesis [3]. Different cell types migrate in opposite directions in EFs [4], and the same cell can switch the directionality depending on conditions [5]. A tug-of-war mechanism between multiple signaling pathways [6] can direct Dictyostelium cells to either cathode or anode. Mechanics of motility is simplest in fish keratocytes, so we turned to keratocytes to investigate their migration in EFs. Keratocytes sense electric fields and migrate to the cathode [7, 8]. Keratocyte fragments [9, 10] are the simplest motile units. Cell fragments from leukocytes are able to respond to chemotactic signals [11], but whether cell fragments are galvanotactic was unknown. We found that keratocyte fragments are the smallest motile electric field-sensing unit: they migrate to the anode, in the opposite direction of whole cells. Myosin II was essential for the direction sensing of fragments but not for parental cells, while PI3 kinase was essential for the direction sensing of whole cells but not for fragments. Thus, two signal transduction pathways, one depending on PI3K, another on myosin, compete to orient motile cells in the electric field. Galvanotaxis is not due to EF force and does not depend on cell or fragment size. We propose a "compass" model according to which protrusive and contractile actomyosin networks self-polarize to the front and rear of the motile cell, respectively, and the electric signal orients both networks toward cathode with different strengths.

  11. Generation of functionally competent and durable engineered blood vessels from human induced pluripotent stem cells.

    Science.gov (United States)

    Samuel, Rekha; Daheron, Laurence; Liao, Shan; Vardam, Trupti; Kamoun, Walid S; Batista, Ana; Buecker, Christa; Schäfer, Richard; Han, Xiaoxing; Au, Patrick; Scadden, David T; Duda, Dan G; Fukumura, Dai; Jain, Rakesh K

    2013-07-30

    Efficient generation of competent vasculogenic cells is a critical challenge of human induced pluripotent stem (hiPS) cell-based regenerative medicine. Biologically relevant systems to assess functionality of the engineered vessels in vivo are equally important for such development. Here, we report a unique approach for the derivation of endothelial precursor cells from hiPS cells using a triple combination of selection markers--CD34, neuropilin 1, and human kinase insert domain-containing receptor--and an efficient 2D culture system for hiPS cell-derived endothelial precursor cell expansion. With these methods, we successfully generated endothelial cells (ECs) from hiPS cells obtained from healthy donors and formed stable functional blood vessels in vivo, lasting for 280 d in mice. In addition, we developed an approach to generate mesenchymal precursor cells (MPCs) from hiPS cells in parallel. Moreover, we successfully generated functional blood vessels in vivo using these ECs and MPCs derived from the same hiPS cell line. These data provide proof of the principle that autologous hiPS cell-derived vascular precursors can be used for in vivo applications, once safety and immunological issues of hiPS-based cellular therapy have been resolved. Additionally, the durability of hiPS-derived blood vessels in vivo demonstrates a potential translation of this approach in long-term vascularization for tissue engineering and treatment of vascular diseases. Of note, we have also successfully generated ECs and MPCs from type 1 diabetic patient-derived hiPS cell lines and use them to generate blood vessels in vivo, which is an important milestone toward clinical translation of this approach.

  12. Phospholipases of Mineralization Competent Cells and Matrix Vesicles: Roles in Physiological and Pathological Mineralizations

    Directory of Open Access Journals (Sweden)

    René Buchet

    2013-03-01

    Full Text Available The present review aims to systematically and critically analyze the current knowledge on phospholipases and their role in physiological and pathological mineralization undertaken by mineralization competent cells. Cellular lipid metabolism plays an important role in biological mineralization. The physiological mechanisms of mineralization are likely to take place in tissues other than in bones and teeth under specific pathological conditions. For instance, vascular calcification in arteries of patients with renal failure, diabetes mellitus or atherosclerosis recapitulates the mechanisms of bone formation. Osteoporosis—a bone resorbing disease—and rheumatoid arthritis originating from the inflammation in the synovium are also affected by cellular lipid metabolism. The focus is on the lipid metabolism due to the effects of dietary lipids on bone health. These and other phenomena indicate that phospholipases may participate in bone remodelling as evidenced by their expression in smooth muscle cells, in bone forming osteoblasts, chondrocytes and in bone resorbing osteoclasts. Among various enzymes involved, phospholipases A1 or A2, phospholipase C, phospholipase D, autotaxin and sphingomyelinase are engaged in membrane lipid remodelling during early stages of mineralization and cell maturation in mineralization-competent cells. Numerous experimental evidences suggested that phospholipases exert their action at various stages of mineralization by affecting intracellular signaling and cell differentiation. The lipid metabolites—such as arachidonic acid, lysophospholipids, and sphingosine-1-phosphate are involved in cell signaling and inflammation reactions. Phospholipases are also important members of the cellular machinery engaged in matrix vesicle (MV biogenesis and exocytosis. They may favour mineral formation inside MVs, may catalyse MV membrane breakdown necessary for the release of mineral deposits into extracellular matrix (ECM, or

  13. Genetic education and the challenge of genomic medicine: development of core competences to support preparation of health professionals in Europe

    DEFF Research Database (Denmark)

    Skirton, Heather; Lewis, Celine; Kent, Alastair;

    2010-01-01

    in professional education and regulation between European countries, setting curricula may not be practical. Core competences are used as a basis for health professional education in many fields and settings. An Expert Group working under the auspices of the EuroGentest project and European Society of Human...... Genetics Education Committee agreed that a pragmatic solution to the need to establish common standards for education and practice in genetic health care was to agree to a set of core competences that could apply across Europe. These were agreed through an exhaustive process of consultation with relevant......The use of genetics and genomics within a wide range of health-care settings requires health professionals to develop expertise to practise appropriately. There is a need for a common minimum standard of competence in genetics for health professionals in Europe but because of differences...

  14. In vitro development competence of bovine nuclear transfer embryos derived from Nanog-overexpressing fibroblast cells

    Directory of Open Access Journals (Sweden)

    Xi-bang Zheng, Yan Yun, Yong-ce Hu, Yong Li, Hua-yan Wang, Xiao-ling Ma, Jin-qiang Sui, An-min Lei and Zhong-ying Dou

    2014-04-01

    Full Text Available The purpose of this study was to establish Nanog-expressing cell lines that can be used as donor cells to construct transgenic cloned embryos, and to investigate their in vitro development competence. By reverse transcription-polymerase chain reaction (RT-PCR, the cDNA of Nanog gene was cloned from fetal bovine primordial genital ridge tissues. The gene was inserted into PMD18-T vector using recombination techniques and then subcloned into vector pEGFP-C1. After confirmation by restrictive endonuclease digestion and sequencing, the recombinant plasmid pEGFP-Nanog was transfected into skin fibroblast cells. A stable transfected cell line was successfully established after two months of selection with neomycine (G418. Fluorescence microscopy, RT-PCR, and Western Blotting assays indicated that Nanog mRNA and EGFP-Nanog fusion protein were expressed in these cells. The EGFP-Nanog expressing fibroblast cells and the intact fibroblast cells (BEF422 were respectively used to construct cloned embryos. The results showed that the cleavage rate of recombinant embryos in BEF422 cells was significantly (P<0.05 higher than in EGFP-Nanog expressing cells (82.14 vs 40.38 %, but the blastocyst development rate in the latter was slightly higher than in the former (17.30 vs 14.29% (P<0.05, indicating that Nanog-overexpressed fibroblasts may be a better candidate of donor cells. To our knowledge, this is the first time that Nanog gene has been introduced into fibroblast cells to produce cloned embryos in bovine.

  15. Propagation of human iPS cells in alginate-based microcapsules prepared using reactions catalyzed by horseradish peroxidase and catalase.

    Science.gov (United States)

    Ashida, Tomoaki; Sakai, Shinji; Taya, Masahito

    2016-09-01

    Cell encapsulation has been investigated as a bioproduction system in the biomedical and pharmaceutical fields. We encaps-ulated human induced pluripotent stem (hiPS) cells in duplex microcapsules prepared from an alginate derivative possessing phenolic hydroxyl moieties, in a single-step procedure based on two competing enzymatic reactions catalyzed by horseradish peroxidase (HRP) and catalase. The encapsulated cells maintained 91.4% viability and proliferated to fill the microcapsules following 19 days of culture. Encapsulated hiPS cells showed pluripotency comparable to that of unencapsulated cells during the cultures, as demonstrated by the expression of the SSEA-4 marker.

  16. Preparation of open-cell metal foams by investment cast

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Metal foams are a new kind of materials with low densities and novel physical, mechanical, thermal, electrical and acoustic properties. They can be divided into closed and open cell structures. In this paper the open cell structures,called sponges, were treated. A new technique to manufacture sponges by plaster investment casting was described.Experimental results show that it is essential to make a sound plaster mould by casting plaster slurry into the polyurethane foams and infiltrate the open channels of the baked plaster mold by molten metal. The optimal processes include plaster slurry preparation, plaster mold baking, and molten metal infiltration. The sponge sample with porosity of 97% is presented.

  17. Preparation of open-cell metal foams by investment cast

    Directory of Open Access Journals (Sweden)

    Lucai WANG

    2005-02-01

    Full Text Available Metal foams are a new kind of materials with low densities and novel physical, mechanical, thermal, electrical and acoustic properties. They can be divided into closed and open cell structures. In this paper the open cell structures, called sponges, were treated. A new technique to manufacture sponges by plaster investment casting was described. Experimental results show that it is essential to make a sound plaster mould by casting plaster slurry into the polyurethane foams and infiltrate the open channels of the baked plaster mold by molten metal. The optimal processes include plaster slurry preparation, plaster mold baking, and molten metal infiltration. The sponge sample with porosity of 97% is presented.

  18. Evidence for different pathways during horizontal gene transfer in competent Bacillus subtilis cells.

    Directory of Open Access Journals (Sweden)

    Dawit Kidane

    2009-09-01

    Full Text Available Cytological and genetic evidence suggests that the Bacillus subtilis DNA uptake machinery localizes at a single cell pole and takes up single-stranded (ss DNA. The integration of homologous donor DNA into the recipient chromosome requires RecA, while plasmid establishment, which is independent of RecA, requires at least RecO and RecU. RecA and RecN colocalize at the polar DNA uptake machinery, from which RecA forms filamentous structures, termed threads, in the presence of chromosomal DNA. We show that the transformation of chromosomal and of plasmid DNA follows distinct pathways. In the absence of DNA, RecU accumulated at a single cell pole in competent cells, dependent on RecA. Upon addition of any kind of DNA, RecA formed highly dynamic thread structures, which rapidly grew and shrank, and RecU dissipated from the pole. RecO visibly accumulated at the cell pole only upon addition of plasmid DNA, and, to a lesser degree, of phage DNA, but not of chromosomal DNA. RecO accumulation was weakly influenced by RecN, but not by RecA. RecO annealed ssDNA complexed with SsbA in vitro, independent of any nucleotide cofactor. The DNA end-joining Ku protein was also found to play a role in viral and plasmid transformation. On the other hand, transfection with SPP1 phage DNA required functions from both chromosomal and plasmid transformation pathways. The findings show that competent bacterial cells possess a dynamic DNA recombination machinery that responds in a differential manner depending if entering DNA shows homology with recipient DNA or has self-annealing potential. Transformation with chromosomal DNA only requires RecA, which forms dynamic filamentous structures that may mediate homology search and DNA strand invasion. Establishment of circular plasmid DNA requires accumulation of RecO at the competence pole, most likely mediating single-strand annealing, and RecU, which possibly down-regulates RecA. Transfection with SPP1 viral DNA follows an

  19. Evaluation of two different dendritic cell preparations with BCG reactivity

    Directory of Open Access Journals (Sweden)

    Fol Marek

    2016-01-01

    Full Text Available Dendritic cells (DCs play a key-role in the immune response against intracellular bacterial pathogens, including mycobacteria. Monocyte-derived dendritic cells (MoDCs are considered to behave as inflammatory cell populations. Different immunomagnetic methods (positive and negative can be used to purify monocytes before their in vitro differentiation and their culture behavior can be expected to be different. In this study we evaluated the reactivity of two dendritic cell populations towards the Bacillus Calmette-Guérin (BCG antigen. Monocytes were obtained from the blood of healthy donors, using positive and negative immunomagnetic separation methods. The expression of DC-SIGN, CD86, CD80, HLA-DR and CD40 on MoDCs was estimated by flow cytometry. The level of IL-12p70, IL-10 and TNF-α was measured by ELISA. Neither of the tested methods affected the surface marker expression of DCs. No significant alteration in immunological response, measured by cytokine production, was noted either. After BCG stimulation, the absence of IL-12, but the IL-23 production was observed in both cell preparations. Positive and negative magnetic separation methods are effective techniques to optimize the preparation of monocytes as the source of MoDCs for potential clinical application.

  20. Retrotransposition-Competent Human LINE-1 Induces Apoptosis in Cancer Cells With Intact p53

    Directory of Open Access Journals (Sweden)

    Abdelali Haoudi

    2004-01-01

    Full Text Available Retrotransposition of human LINE-1 (L1 element, a major representative non-LTR retrotransposon in the human genome, is known to be a source of insertional mutagenesis. However, nothing is known about effects of L1 retrotransposition on cell growth and differentiation. To investigate the potential for such biological effects and the impact that human L1 retrotransposition has upon cancer cell growth, we examined a panel of human L1 transformed cell lines following a complete retrotransposition process. The results demonstrated that transposition of L1 leads to the activation of the p53-mediated apoptotic pathway in human cancer cells that possess a wild-type p53. In addition, we found that inactivation of p53 in cells, where L1 was undergoing retrotransposition, inhibited the induction of apoptosis. This suggests an association between active retrotransposition and a competent p53 response in which induction of apoptosis is a major outcome. These data are consistent with a model in which human retrotransposition is sensed by the cell as a “genetic damaging event” and that massive retrotransposition triggers signaling pathways resulting in apoptosis.

  1. Competency-Based Framework and Continuing Education for Preparing a Skilled School Health Workforce for Asthma Care: The Colorado Experience

    Science.gov (United States)

    Cicutto, Lisa; Gleason, Melanie; Haas-Howard, Christy; Jenkins-Nygren, Lynn; Labonde, Susan; Patrick, Kathy

    2017-01-01

    School health teams commonly address the needs of students with asthma, which requires specific knowledge and skills. To develop a skilled school health team, a competency-based framework for managing asthma in schools was developed. A modified Delphi with 31 panelists was completed. Consensus (=80% agreement) was reached for all 148 items…

  2. Pre-Service Chemistry Teachers' Competencies in the Laboratory: A Cross-Grade Study in Solution Preparation

    Science.gov (United States)

    Karatas, F. O.

    2016-01-01

    One of the prerequisites for chemistry teacher candidates is to demonstrate certain laboratory skills. This article aims to determine and discuss the competencies of pre-service chemistry teachers in a chemistry laboratory context working with solution chemistry content. The participants in this study consisted of a group of pre-service chemistry…

  3. Preparing the health workforce in Ethiopia : A Cross-sectional study of competence of anesthesia graduating students

    NARCIS (Netherlands)

    Kibwana, Sharon; Woldemariam, Damtew; Misganaw, Awoke; Teshome, Mihereteab; Akalu, Leulayehu; Kols, Adrienne; Kim, Young Mi; Mengistu, Samuel; van Roosmalen, Jos; Stekelenburg, Jelle

    2016-01-01

    BACKGROUND: Efforts to address shortages of health workers in low-resource settings have focused on rapidly increasing the number of higher education programs for health workers. This study examines selected competencies achieved by graduating Bachelor of Science and nurse anesthetist students in

  4. Preparing the health workforce in Ethiopia : A Cross-sectional study of competence of anesthesia graduating students

    NARCIS (Netherlands)

    Kibwana, Sharon; Woldemariam, Damtew; Misganaw, Awoke; Teshome, Mihereteab; Akalu, Leulayehu; Kols, Adrienne; Kim, Young Mi; Mengistu, Samuel; van Roosmalen, Jos; Stekelenburg, Jelle

    2016-01-01

    BACKGROUND: Efforts to address shortages of health workers in low-resource settings have focused on rapidly increasing the number of higher education programs for health workers. This study examines selected competencies achieved by graduating Bachelor of Science and nurse anesthetist students in Et

  5. Special Education "Preparation Gap": Perceptions of the Western Region Special Education Advisers Concerning Pennsylvania's Special Education Competencies for School Leaders

    Science.gov (United States)

    Conner, Matthew Richard

    2012-01-01

    This study determines the strengths and weaknesses of the Special Education Competencies for School Leaders framework as adopted by the Pennsylvania Department of Education. The perspectives of Special Education Advisers (individuals with significant experience in serving students with disabilities) are gathered through semi-structured interviews.…

  6. Competence is Competence

    DEFF Research Database (Denmark)

    Bramming, Pia

    2004-01-01

    The article will address competence, its' diffusion, application, and the consequence of this application within the field of Human Resource Management (HRM). The concept competence-in-practice will be presented and in conclusion the article will consider implications and possibilities of compete...

  7. Preparation of pancreatic β-cells from human iPS cells with small molecules.

    Science.gov (United States)

    Hosoya, Masaki

    2012-01-01

    Human induced pluripotent stem (iPS) cells obtained from patients are expected to be a useful source for cell transplantation therapy, because many patients (including those with type 1 diabetes and severe type 2 diabetes) are on waiting lists for transplantation for a long time due to the shortage of donors. At present, many concerns related to clinical application of human iPS cells have been raised, but rapid development of methods for the establishment, culture, and standardization of iPS cells will lead autologous cell therapy to be realistic sooner or later. However, establishment of a method for preparing some of desired cell types is still challenging. Regarding pancreatic β-cells, there have been many reports about differentiation of these cells from human embryonic stem (ES)/iPS cells, but a protocol for clinical application has still not been established. Since there is clear proof that cell transplantation therapy is effective for diabetes based on the results of clinical islet transplantation, pancreatic β-cells prepared from human iPS cells are considered likely to be effective for reducing the burden on patients. In this article, the current status of procedures for preparing pancreatic β-cells from human ES/iPS cells, including effective use of small molecules, is summarized, and some of the problems that still need to be overcome are discussed.

  8. In Vitro Epigenetic Reprogramming of Human Cardiac Mesenchymal Stromal Cells into Functionally Competent Cardiovascular Precursors

    Science.gov (United States)

    Vecellio, Matteo; Meraviglia, Viviana; Nanni, Simona; Barbuti, Andrea; Scavone, Angela; DiFrancesco, Dario; Farsetti, Antonella; Pompilio, Giulio; Colombo, Gualtiero I.; Capogrossi, Maurizio C.

    2012-01-01

    Adult human cardiac mesenchymal-like stromal cells (CStC) represent a relatively accessible cell type useful for therapy. In this light, their conversion into cardiovascular precursors represents a potential successful strategy for cardiac repair. The aim of the present work was to reprogram CStC into functionally competent cardiovascular precursors using epigenetically active small molecules. CStC were exposed to low serum (5% FBS) in the presence of 5 µM all-trans Retinoic Acid (ATRA), 5 µM Phenyl Butyrate (PB), and 200 µM diethylenetriamine/nitric oxide (DETA/NO), to create a novel epigenetically active cocktail (EpiC). Upon treatment the expression of markers typical of cardiac resident stem cells such as c-Kit and MDR-1 were up-regulated, together with the expression of a number of cardiovascular-associated genes including KDR, GATA6, Nkx2.5, GATA4, HCN4, NaV1.5, and α-MHC. In addition, profiling analysis revealed that a significant number of microRNA involved in cardiomyocyte biology and cell differentiation/proliferation, including miR 133a, 210 and 34a, were up-regulated. Remarkably, almost 45% of EpiC-treated cells exhibited a TTX-sensitive sodium current and, to a lower extent in a few cells, also the pacemaker If current. Mechanistically, the exposure to EpiC treatment introduced global histone modifications, characterized by increased levels of H3K4Me3 and H4K16Ac, as well as reduced H4K20Me3 and H3s10P, a pattern compatible with reduced proliferation and chromatin relaxation. Consistently, ChIP experiments performed with H3K4me3 or H3s10P histone modifications revealed the presence of a specific EpiC-dependent pattern in c-Kit, MDR-1, and Nkx2.5 promoter regions, possibly contributing to their modified expression. Taken together, these data indicate that CStC may be epigenetically reprogrammed to acquire molecular and biological properties associated with competent cardiovascular precursors. PMID:23284745

  9. In vitro epigenetic reprogramming of human cardiac mesenchymal stromal cells into functionally competent cardiovascular precursors.

    Directory of Open Access Journals (Sweden)

    Matteo Vecellio

    Full Text Available Adult human cardiac mesenchymal-like stromal cells (CStC represent a relatively accessible cell type useful for therapy. In this light, their conversion into cardiovascular precursors represents a potential successful strategy for cardiac repair. The aim of the present work was to reprogram CStC into functionally competent cardiovascular precursors using epigenetically active small molecules. CStC were exposed to low serum (5% FBS in the presence of 5 µM all-trans Retinoic Acid (ATRA, 5 µM Phenyl Butyrate (PB, and 200 µM diethylenetriamine/nitric oxide (DETA/NO, to create a novel epigenetically active cocktail (EpiC. Upon treatment the expression of markers typical of cardiac resident stem cells such as c-Kit and MDR-1 were up-regulated, together with the expression of a number of cardiovascular-associated genes including KDR, GATA6, Nkx2.5, GATA4, HCN4, NaV1.5, and α-MHC. In addition, profiling analysis revealed that a significant number of microRNA involved in cardiomyocyte biology and cell differentiation/proliferation, including miR 133a, 210 and 34a, were up-regulated. Remarkably, almost 45% of EpiC-treated cells exhibited a TTX-sensitive sodium current and, to a lower extent in a few cells, also the pacemaker I(f current. Mechanistically, the exposure to EpiC treatment introduced global histone modifications, characterized by increased levels of H3K4Me3 and H4K16Ac, as well as reduced H4K20Me3 and H3s10P, a pattern compatible with reduced proliferation and chromatin relaxation. Consistently, ChIP experiments performed with H3K4me3 or H3s10P histone modifications revealed the presence of a specific EpiC-dependent pattern in c-Kit, MDR-1, and Nkx2.5 promoter regions, possibly contributing to their modified expression. Taken together, these data indicate that CStC may be epigenetically reprogrammed to acquire molecular and biological properties associated with competent cardiovascular precursors.

  10. Diego and Prickle regulate Frizzled planar cell polarity signalling by competing for Dishevelled binding.

    Science.gov (United States)

    Jenny, Andreas; Reynolds-Kenneally, Jessica; Das, Gishnu; Burnett, Micheal; Mlodzik, Marek

    2005-07-01

    Epithelial planar cell polarity (PCP) is evident in the cellular organization of many tissues in vertebrates and invertebrates. In mammals, PCP signalling governs convergent extension during gastrulation and the organization of a wide variety of structures, including the orientation of body hair and sensory hair cells of the inner ear. In Drosophila melanogaster, PCP is manifest in adult tissues, including ommatidial arrangement in the compound eye and hair orientation in wing cells. PCP establishment requires the conserved Frizzled/Dishevelled PCP pathway. Mutations in PCP-pathway-associated genes cause aberrant orientation of body hair or inner-ear sensory cells in mice, or misorientation of ommatidia and wing hair in D. melanogaster. Here we provide mechanistic insight into Frizzled/Dishevelled signalling regulation. We show that the ankyrin-repeat protein Diego binds directly to Dishevelled and promotes Frizzled signalling. Dishevelled can also be bound by the Frizzled PCP antagonist Prickle. Strikingly, Diego and Prickle compete with one another for Dishevelled binding, thereby modulating Frizzled/Dishevelled activity and ensuring tight control over Frizzled PCP signalling.

  11. Development of a replication-competent lentivirus assay for dendritic cell-targeting lentiviral vectors

    Directory of Open Access Journals (Sweden)

    Daniel C Farley

    2015-01-01

    Full Text Available It is a current regulatory requirement to demonstrate absence of detectable replication-competent lentivirus (RCL in lentiviral vector products prior to use in clinical trials. Immune Design previously described an HIV-1-based integration-deficient lentiviral vector for use in cancer immunotherapy (VP02. VP02 is enveloped with E1001, a modified Sindbis virus glycoprotein which targets dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN expressed on dendritic cells in vivo. Vector enveloped with E1001 does not transduce T-cell lines used in standard HIV-1-based RCL assays, making current RCL testing formats unsuitable for testing VP02. We therefore developed a novel assay to test for RCL in clinical lots of VP02. This assay, which utilizes a murine leukemia positive control virus and a 293F cell line expressing the E1001 receptor DC-SIGN, meets a series of evaluation criteria defined in collaboration with US regulatory authorities and demonstrates the ability of the assay format to amplify and detect a hypothetical RCL derived from VP02 vector components. This assay was qualified and used to test six independent GMP production lots of VP02, in which no RCL was detected. We propose that the evaluation criteria used to rationally design this novel method should be considered when developing an RCL assay for any lentiviral vector.

  12. Preparation of carboxymethyl cellulose based microgels for cell encapsulation

    Directory of Open Access Journals (Sweden)

    Y. Ke

    2014-11-01

    Full Text Available Biocompatible and biodegradable carboxymethyl cellulose (CMC has been modified with 4-hydroxybenzylamine (CMC-Ph in order to prepare CMC-based microgels through the horseradish peroxidise/hydrogen peroxide enzymatic reaction. CMC-Ph was identified as a blend, and the amount of the grafted 4-hydroxybenzylamine per 100 units of CMC was between 17 and 23 according to the molecular weight of CMC. Through a special designed co-flowing microfluidic device, CMC-Ph microgels were prepared with the radius from 100 to 500 μm via adjusting the flow rates of the disperse phase and the continuous phase, respectively. The chondrocytic cell line ATDC5 was encapsulated in the CMC-Ph microgels. The cell-laden microgels were cultured for up to 40 days, illustrating the biocompatibility of CMC-Ph and the microfluidic approach through the enzymatic crosslinking reaction primarily. CMC-Ph showed a great promise to encapsulate the cells for further fabrication of the injectable scaffolds.

  13. Competencies for midwifery teachers.

    Science.gov (United States)

    Thompson, Joyce E

    2002-12-01

    Saving women's lives with cost-quality effective midwifery care is based on sound pre-service and ongoing education. Effective midwifery education requires competent, caring, and compassionate teachers. In this paper, I address the basic competencies required of midwives who teach others to be midwives. These competencies are important regardless of level of student taught, type of educational programme, or number of years of midwifery experience that learners bring to the educational setting. The competencies are based on the midwifery philosophy, values and model of care. Competent midwifery teachers must be competent midwifery clinicians for their primary role is to set the boundaries of safety for each level of learner. Formal preparation for teaching, understanding how adults learn, understanding how to develop an appropriate plan for learning (curriculum), and developing competency in a variety of teaching methods for both theory and clinical practice are included in the competencies discussed in this paper.

  14. Long Noncoding RNA HOTAIR Controls Cell Cycle by Functioning as a Competing Endogenous RNA in Esophageal Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Kewei Ren

    2016-12-01

    Full Text Available Recent studies have shown that long noncoding RNAs (lncRNAs play pivotal roles in the initiation and progression of cancer, including esophageal squamous cell carcinoma (ESCC. The lncRNA HOX transcript antisense RNA (HOTAIR was reported to be dysregulated and correlated with the progression of ESCC. However, the biological role and the underlying mechanism of HOTAIR in the development of ESCC remain unclear. Herein, we found that HOTAIR was aberrantly upregulated in ESCC cells and that HOTAIR depletion inhibited proliferation and led to G1 cell cycle arrest in ESCC cells. Besides, we found that HOTAIR acted as an endogenous sponge to downregulate miR-1 expression by directly binding to miR-1. Furthermore, HOTAIR overturned the effect of miR-1 on the proliferation and cell cycle profile in ESCC cells, which involved the derepression of cyclin D1 (CCND1 expression, a target of miR-1. Taken together, our study elucidated a novel HOTAIR /miR-1/CCND1 regulatory axis in which HOTAIR acted as a competing endogenous RNA by sponging miR-1 and upregulated CCND1 expression, thereby facilitating the tumorigenesis of ESCC. Investigation of this lncRNA/miRNA/mRNA pathway may contribute to a better understanding of ESCC pathogenesis and facilitate the development of lncRNA-directed therapy against this disease.

  15. Evaluation of porcine stem cell competence for somatic cell nuclear transfer and production of cloned animals.

    Science.gov (United States)

    Secher, Jan O; Liu, Ying; Petkov, Stoyan; Luo, Yonglun; Li, Dong; Hall, Vanessa J; Schmidt, Mette; Callesen, Henrik; Bentzon, Jacob F; Sørensen, Charlotte B; Freude, Kristine K; Hyttel, Poul

    2017-03-01

    Porcine somatic cell nuclear transfer (SCNT) has been used extensively to create genetically modified pigs, but the efficiency of the methodology is still low. It has been hypothesized that pluripotent or multipotent stem cells might result in increased SCNT efficacy as these cells are closer than somatic cells to the epigenetic state found in the blastomeres and therefore need less reprogramming. Our group has worked with porcine SCNT during the last 20 years and here we describe our experience with SCNT of 3 different stem cell lines. The porcine stem cells used were: Induced pluripotent stem cells (iPSCs) created by lentiviral doxycycline-dependent reprogramming and cultered with a GSK3β- and MEK-inhibitor (2i) and leukemia inhibitor factor (LIF) (2i LIF DOX-iPSCs), iPSCs created by a plasmid-based reprogramming and cultured with 2i and fibroblast growth factor (FGF) (2i FGF Pl-iPSCs) and embryonic germ cells (EGCs), which have earlier been characterized as being multipotent. The SCNT efficiencies of these stem cell lines were compared with that of the two fibroblast cell lines from which the iPSC lines were derived. The blastocyst rates for the 2i LIF DOX-iPSCs were 14.7%, for the 2i FGF Pl-iPSC 10.1%, and for the EGCs 34.5% compared with the fibroblast lines yielding 36.7% and 25.2%. The fibroblast- and EGC-derived embryos were used for embryo transfer and produced live offspring at similar low rates of efficiency (3.2 and 4.0%, respectively) and with several instances of malformations. In conclusion, potentially pluripotent porcine stem cells resulted in lower rates of embryonic development upon SCNT than multipotent stem cells and differentiated somatic cells.

  16. Perovskite/polymer solar cells prepared using solution process

    Science.gov (United States)

    Rosa, E. S.; Shobih; Nursam, N. M.; Saputri, D. G.

    2016-11-01

    We report a simple solution-based process to fabricate a perovskite/polymer tandem solar cell using inorganic CH3NH3PM3 as an absorber and organic PCBM (6,6 phenyl C61- butyric acid methyl ester) as an electron transport layer. The absorber solution was prepared by mixing the CH3NH3I (methyl ammonium iodide) with PbI2 (lead iodide) in DMF (N,N- dimethyl formamide) solvent. The absorber and electron transport layer were deposited by spin coating method. The electrical characteristics generated from the cell under 50 mW/cm2 at 25 °C comprised of an open circuit voltage of 0.31 V, a short circuit current density of 2.53 mA/cm2, and a power conversion efficiency of 0.42%.

  17. COMPETENCY APPROACH USING IN THE ORGANIZATION OF EMPLOYMENT ON PHYSICAL PREPARATION OF CADETS AND STUDENTS OF EDUCATIONAL INSTITUTIONS OF RUSSIAN MINISTRY OF INTERNAL AFFAIRS

    Directory of Open Access Journals (Sweden)

    A. S. PRYAKHIN

    2016-01-01

    Full Text Available Educational institutions of higher education have high demands of familiarizing of cadets and listeners to the values of world and national culture, the formation of skills of effective social interaction, personal development and self-improvement. Physical training of police officers is a determining factor for their effective professional activity. In this regard, the definition of the main approaches to be used in the organization of employment on physical preparation of cadets and students of educational institutions Ministry of Internal Affairs of Russia is a priority. This paper examines the main approaches used in the organization of employment on physical training of cadets and students of educational institutions Ministry of Internal Affairs of Russia. A special place in this system is given the competence approach. In article substantiates the importance competency approach in the organization of employment on physical preparation of the profile of the university. The paper presents the basic guidelines for the organization of "physical training" in educational institutions Ministry of Internal Affairs of Russia. The implementation of modern requirements for training of cadets and students of educational institutions MIA of Russia assumes the achievement of an integrated end result of education, for which can be seen Maturity graduates of professionally significant competences as the unity of generalized knowledge and skills, universal capacity and preparedness to address large groups of tasks - from personal to the social and professional. It is proved that, to use the competency approach to the process of the organization of physical training of cadets and students of educational institutions Russian Ministry of Internal Affairs improves the efficiency of the overall training of the future professionals of the Interior.

  18. A protodermal miR394 signal defines a region of stem cell competence in the Arabidopsis shoot meristem.

    Science.gov (United States)

    Knauer, Steffen; Holt, Anna L; Rubio-Somoza, Ignacio; Tucker, Elise J; Hinze, Annika; Pisch, Melanie; Javelle, Marie; Timmermans, Marja C; Tucker, Matthew R; Laux, Thomas

    2013-01-28

    A long-standing question in plants and animals is how spatial patterns are maintained within stem cell niches despite ongoing cell divisions. Here we address how, during shoot meristem formation in Arabidopsis thaliana, the three apical cell layers acquire stem cell identity. Using a sensitized mutant screen, we identified miR394 as a mobile signal produced by the surface cell layer (the protoderm) that confers stem cell competence to the distal meristem by repressing the F box protein LEAF CURLING RESPONSIVENESS. This repression is required to potentiate signaling from underneath the stem cells by the transcription factor WUSCHEL, maintaining stem cell pluripotency. The interaction of two opposing signaling centers provides a mechanistic framework of how stem cells are localized at the tip of the meristem. Although the constituent cells change, the surface layer provides a stable point of reference in the self-organizing meristem. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Preparation

    Directory of Open Access Journals (Sweden)

    M.M. Dardir

    2014-03-01

    Full Text Available Some hexanamide-mono and di-linoleniate esters were prepared by the reaction of linolenic acid and hexanamide (derived from the reaction of hexanoic acid and diethanolamine. The chemical structure for the newly prepared hexanamide-mono and di-linoleniate esters were elucidated using elemental analysis, (FTIR, H 1NMR and chemical ionization mass spectra (CI/Ms spectroscopic techniques. The results of the spectroscopic analysis indicated that they were prepared through the right method and they have high purity. The new prepared esters have high biodegradability and lower toxicity (environmentally friendly so they were evaluated as a synthetic-based mud (ester-based mud for oil-well drilling fluids. The evaluation included study of the rheological properties, filtration and thermal properties of the ester based-muds formulated with the newly prepared esters compared to the reference commercial synthetic-based mud.

  20. TRAIL causes deletions at the HPRT and TK1 loci of clonogenically competent cells

    Energy Technology Data Exchange (ETDEWEB)

    Miles, Mark A.; Shekhar, Tanmay M. [Department of Biochemistry and Genetics, La Trobe University, Bundoora, Victoria (Australia); La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria (Australia); Hall, Nathan E. [La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria (Australia); Life Sciences Computation Centre, Victorian Life Sciences Computation Initiative, Melbourne, Victoria (Australia); Hawkins, Christine J., E-mail: c.hawkins@latrobe.edu.au [Department of Biochemistry and Genetics, La Trobe University, Bundoora, Victoria (Australia); La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria (Australia)

    2016-05-15

    Highlights: • Treatment with TRAIL or EMS provokes mutations in clonogenically viable TK6 cells. • TRAIL is 2–5-fold less mutagenic than an equivalently lethal concentration of EMS. • EMS mainly causes transition mutations at the HPRT and TK1 loci of TK6 cells. • Most loss-of-function HPRT or TK1 mutations caused by TRAIL treatment are deletions. - Abstract: When chemotherapy and radiotherapy are effective, they function by inducing DNA damage in cancerous cells, which respond by undergoing apoptosis. Some adverse effects can result from collateral destruction of non-cancerous cells, via the same mechanism. Therapy-related cancers, a particularly serious adverse effect of anti-cancer treatments, develop due to oncogenic mutations created in non-cancerous cells by the DNA damaging therapies used to eliminate the original cancer. Physiologically achievable concentrations of direct apoptosis inducing anti-cancer drugs that target Bcl-2 and IAP proteins possess negligible mutagenic activity, however death receptor agonists like TRAIL/Apo2L can provoke mutations in surviving cells, probably via caspase-mediated activation of the nuclease CAD. In this study we compared the types of mutations sustained in the HPRT and TK1 loci of clonogenically competent cells following treatment with TRAIL or the alkylating agent ethyl methanesulfonate (EMS). As expected, the loss-of-function mutations in the HPRT or TK1 loci triggered by exposure to EMS were almost all transitions. In contrast, only a minority of the mutations identified in TRAIL-treated clones lacking HPRT or TK1 activity were substitutions. Almost three quarters of the TRAIL-induced mutations were partial or complete deletions of the HPRT or TK1 genes, consistent with sub-lethal TRAIL treatment provoking double strand breaks, which may be mis-repaired by non-homologous end joining (NHEJ). Mis-repair of double-strand breaks following exposure to chemotherapy drugs has been implicated in the pathogenesis of

  1. The second Summer Youth Olympic Games in Nanjing, People's Republic of China: preparing youth athletes to compete in the heat

    Directory of Open Access Journals (Sweden)

    Brito J

    2014-09-01

    Full Text Available Joao Brito,1 Sebastien Racinais,2 George P Nassis,1,3 1National Sports Medicine Programme, Excellence in Football Project, Aspetar – Qatar Orthopaedic and Sports Medicine Hospital, Doha, Qatar; 2Athlete Health and Performance Research Centre, Aspetar – Qatar Orthopaedic and Sports Medicine Hospital, Doha, Qatar; 3School of Physical Education and Sport Science, National and Kapodistrian University of Athens, Athens, Greece  All authors contributed equally to this manuscript Abstract: The second Summer Youth Olympic Games will take place August 16–28, 2014 in Nanjing, People's Republic of China during the peak of the summer. Nanjing has been reported as one of the hottest cities in the People's Republic of China, with temperatures reaching as high as 40°C. There is limited clinical evidence of the real risks that youth athletes face when training and competing in the heat, but some recommendations can be made. The estimated average wet bulb globe temperature for Nanjing in August is 32°C, which has been classified as a very high risk/stop play condition for heat illness and injury. Training and competing under extreme heat conditions could stress the thermoregulatory system and adversely affect health and performance. However, current guidelines appear to be inadequate or too conservative, and mostly focus on adult elite athletes. Therefore, proper preventive measures are warranted to reduce the risks of heat illness and injury. With proper heat acclimatization and monitoring, youth athletes can exercise reasonably well and safely in the heat. During the second Summer Youth Olympic Games in Nanjing, People's Republic of China, special attention should be devoted to athletes exposed to long and extensive sunny and hot conditions. Keywords: youth sports, heat acclimatization, thermoregulation

  2. The Perceived Benefits of a Preparing Future Faculty Program and Its Effect on Job Satisfaction, Confidence, and Competence

    Science.gov (United States)

    Wurgler, Emily; VanHeuvelen, Jane S.; Rohrman, Shawna; Loehr, Annalise; Grace, Matthew K.

    2014-01-01

    The training of effective instructors and future faculty members is a critical component of doctoral programs in sociology. Many universities and departments have instituted a single course, course sequence, or certification program dedicated to the preparation of future academic faculty. This article evaluates the efficacy of one such program,…

  3. Effect of Perovskite Film Preparation on Performance of Solar Cells

    Directory of Open Access Journals (Sweden)

    Yaxian Pei

    2016-01-01

    Full Text Available For the perovskite solar cells (PSCs, the performance of the PSCs has become the focus of the research by improving the crystallization and morphology of the perovskite absorption layer. In this thesis, based on the structure of mesoporous perovskite solar cells (MPSCs, we designed the experiments to improve the photovoltaic performance of the PSCs by improved processing technique, which mainly includes the following two aspects. Before spin-coating PbI2 solution, we control the substrate temperature to modify the crystal quality and morphology of perovskite films. On the other hand, before annealing, we keep PbI2 films for the different drying time at room temperature to optimize films morphology. In our trials, it was found that the substrate temperature is more important in determining the photovoltaic performance than drying time. These results indicate that the crystallization and morphology of perovskite films affect the absorption intensity and obviously influence the short circuit current density of MPSCs. Utilizing films prepared by mentioning two methods, MPSCs with maximum power conversion efficiency of over 4% were fabricated for the active area of 0.5 × 0.5 cm2.

  4. Human NK cells of mice with reconstituted human immune system components require preactivation to acquire functional competence.

    Science.gov (United States)

    Strowig, Till; Chijioke, Obinna; Carrega, Paolo; Arrey, Frida; Meixlsperger, Sonja; Rämer, Patrick C; Ferlazzo, Guido; Münz, Christian

    2010-11-18

    To investigate human natural killer (NK)-cell reactivity in vivo we have reconstituted human immune system components by transplantation of human hematopoietic progenitor cells into NOD-scid IL2Rγ(null) mice. We demonstrate here that this model allows the development of all NK-cell subsets that are also found in human adult peripheral and cord blood, including NKp46(+)CD56(-) NK cells. Similar to human cord blood, NK cells from these reconstituted mice require preactivation by interleukin-15 to reach the functional competence of human adult NK cells. Mainly the terminally differentiated CD16(+) NK cells demonstrate lower reactivity without this stimulation. After preactivation, both CD16(+) and CD16(-) NK cells efficiently produce interferon-γ and degranulate in response to stimulation with NK cell-susceptible targets, including K562 erythroleukemia cells. NK-cell lines, established from reconstituted mice, demonstrate cytotoxicity against this tumor cell line. Importantly, preactivation can as well be achieved by bystander cell maturation via poly I:C stimulation in vitro and injection of this maturation stimulus in vivo. Preactivation in vivo enhances killing of human leukocyte antigen class I negative tumor cells after their adoptive transfer. These data suggest that a functional, but resting, NK-cell compartment can be established in immune-compromised mice after human hematopoietic progenitor cell transfer.

  5. PREPARATION AND CHARACTERIZATION OF SOLID ELECTROLYTES: FUEL CELL APPLICATIONS

    Energy Technology Data Exchange (ETDEWEB)

    Rambabu Bobba; Josef Hormes; T. Wang; Jaymes A. Baker; Donald G. Prier; Tommy Rockwood; Dinesha Hawkins; Saleem Hasan; V. Rayanki

    1997-12-31

    The intent of this project with Federal Energy Technology Center (FETC)/Morgantown Energy Technology Center (METC) is to develop research infrastructure conductive to Fuel Cell research at Southern University and A and M College, Baton Route. A state of the art research laboratory (James Hall No.123 and No.114) for energy conversion and storage devices was developed during this project duration. The Solid State Ionics laboratory is now fully equipped with materials research instruments: Arbin Battery Cycling and testing (8 channel) unit, Electrochemical Analyzer (EG and G PAR Model 273 and Solartron AC impedance analyzer), Fuel Cell test station (Globe Tech), Differential Scanning Calorimeter (DSC-10), Thermogravimetric Analyzer (TGA), Scanning Tunneling Microscope (STM), UV-VIS-NIR Absorption Spectrometer, Fluorescence Spectrometer, FT-IR Spectrometer, Extended X-ray Absorption Fine Structure (EXAFS) measurement capability at Center for Advanced Microstructure and Devices (CAMD- a multimillion dollar DOE facility), Glove Box, gas hood chamber, high temperature furnaces, hydraulic press and several high performance computers. IN particular, a high temperature furnace (Thermodyne 6000 furnace) and a high temperature oven were acquired through this project funds. The PI Dr. R Bobba has acquired additional funds from federal agencies include NSF-Academic Research Infrastructure program and other DOE sites. They have extensively used the multimillion dollar DOE facility ''Center'' for Advanced Microstructures and Devices (CAMD) for electrochemical research. The students were heavily involved in the experimental EXAFS measurements and made use of their DCM beamline for EXAFS research. The primary objective was to provide hands on experience to the selected African American undergraduate and graduate students in experimental energy research.The goal was to develop research skills and involve them in the Preparation and Characterization of Solid

  6. Preparation courses for medical clerkships and the final clinical internship in medical education – The Magdeburg Curriculum for Healthcare Competence

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    Spura, Anke

    2016-05-01

    Full Text Available Background/Goals: Supporting medical students entering their internships – the clinical clerkship and the internship “final clinical year” (Praktisches Jahr, PJ – the seminars “Ready for Clerkship” and “Ready for PJ” were held for the first time in 2014 and continued successfully in 2015. These seminars are part of the “Magdeburg Curriculum for Healthcare Competence” (Magdeburger Curriculum zur Versorgungskompetenz, MCV. The concept comprises three main issues: “Understanding interdisciplinary clinical procedures”, “Interprofessional collaboration”, and “Individual cases and their reference to the system.” The aim of the seminar series is to prepare students as medical trainees for their role in the practice-oriented clinical clerkship and PJ, respectively.Methods: Quality assurance evaluations and didactic research are integral parts of the seminars. In preparation for the “Ready for PJ” seminar a needs assessment was conducted. The seminars were rated by the participants using an anonymized questionnaire consisting of a 5-choice Likert scale (ranging from 1=fully agree to 5=fully disagree and spaces for comments that was generated by the evaluation software Evasys.Results: The results are presented for the preparatory seminars “Ready for Clerkship” and “Fit für PJ” held in 2014 and 2015. Overall, the students regarded the facultative courses as very good preparation for the clerkship as well as for the PJ. The three-dimensional main curricular concept of the MCV was recognized in the evaluation as a valuable educational approach. Interprofessional collaboration, taught by instructors focussing in teamwork between disciplines, was scored positively and highly valued.Conclusions: The “Magdeburg Curriculum for Healthcare Competence” (MCV integrates clerkship and PJ in a framing educational concept and allows students a better appreciation of their role in patient care and the tasks that they will

  7. Integration of signals along orthogonal axes of the vertebrate neural tube controls progenitor competence and increases cell diversity.

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    Noriaki Sasai

    2014-07-01

    Full Text Available A relatively small number of signals are responsible for the variety and pattern of cell types generated in developing embryos. In part this is achieved by exploiting differences in the concentration or duration of signaling to increase cellular diversity. In addition, however, changes in cellular competence-temporal shifts in the response of cells to a signal-contribute to the array of cell types generated. Here we investigate how these two mechanisms are combined in the vertebrate neural tube to increase the range of cell types and deliver spatial control over their location. We provide evidence that FGF signaling emanating from the posterior of the embryo controls a change in competence of neural progenitors to Shh and BMP, the two morphogens that are responsible for patterning the ventral and dorsal regions of the neural tube, respectively. Newly generated neural progenitors are exposed to FGF signaling, and this maintains the expression of the Nk1-class transcription factor Nkx1.2. Ventrally, this acts in combination with the Shh-induced transcription factor FoxA2 to specify floor plate cells and dorsally in combination with BMP signaling to induce neural crest cells. As development progresses, the intersection of FGF with BMP and Shh signals is interrupted by axis elongation, resulting in the loss of Nkx1.2 expression and allowing the induction of ventral and dorsal interneuron progenitors by Shh and BMP signaling to supervene. Hence a similar mechanism increases cell type diversity at both dorsal and ventral poles of the neural tube. Together these data reveal that tissue morphogenesis produces changes in the coincidence of signals acting along orthogonal axes of the neural tube and this is used to define spatial and temporal transitions in the competence of cells to interpret morphogen signaling.

  8. Integration of signals along orthogonal axes of the vertebrate neural tube controls progenitor competence and increases cell diversity.

    Science.gov (United States)

    Sasai, Noriaki; Kutejova, Eva; Briscoe, James

    2014-07-01

    A relatively small number of signals are responsible for the variety and pattern of cell types generated in developing embryos. In part this is achieved by exploiting differences in the concentration or duration of signaling to increase cellular diversity. In addition, however, changes in cellular competence-temporal shifts in the response of cells to a signal-contribute to the array of cell types generated. Here we investigate how these two mechanisms are combined in the vertebrate neural tube to increase the range of cell types and deliver spatial control over their location. We provide evidence that FGF signaling emanating from the posterior of the embryo controls a change in competence of neural progenitors to Shh and BMP, the two morphogens that are responsible for patterning the ventral and dorsal regions of the neural tube, respectively. Newly generated neural progenitors are exposed to FGF signaling, and this maintains the expression of the Nk1-class transcription factor Nkx1.2. Ventrally, this acts in combination with the Shh-induced transcription factor FoxA2 to specify floor plate cells and dorsally in combination with BMP signaling to induce neural crest cells. As development progresses, the intersection of FGF with BMP and Shh signals is interrupted by axis elongation, resulting in the loss of Nkx1.2 expression and allowing the induction of ventral and dorsal interneuron progenitors by Shh and BMP signaling to supervene. Hence a similar mechanism increases cell type diversity at both dorsal and ventral poles of the neural tube. Together these data reveal that tissue morphogenesis produces changes in the coincidence of signals acting along orthogonal axes of the neural tube and this is used to define spatial and temporal transitions in the competence of cells to interpret morphogen signaling.

  9. Sustained ERK Activation Underlies Reprogramming in Regeneration-Competent Salamander Cells and Distinguishes Them from Their Mammalian Counterparts

    OpenAIRE

    Maximina H. Yun; Phillip B. Gates; Jeremy P. Brockes

    2014-01-01

    Summary In regeneration-competent vertebrates, such as salamanders, regeneration depends on the ability of various differentiated adult cell types to undergo natural reprogramming. This ability is rarely observed in regeneration-incompetent species such as mammals, providing an explanation for their poor regenerative potential. To date, little is known about the molecular mechanisms mediating natural reprogramming during regeneration. Here, we have identified the extent of extracellular signa...

  10. Germ cell transplantation using sexually competent fish: an approach for rapid propagation of endangered and valuable germlines.

    Directory of Open Access Journals (Sweden)

    Sullip K Majhi

    Full Text Available The transplantation of germ cells into adult recipient gonads is a tool with wide applications in animal breeding and conservation of valuable and/or endangered species; it also provides a means for basic studies involving germ cell (GC proliferation and differentiation. Here we describe the establishment of a working model for xenogeneic germ cell transplantation (GCT in sexually competent fish. Spermatogonial cells isolated from juveniles of one species, the pejerrey Odontesthes bonariensis (Atherinopsidae, were surgically transplanted into the gonads of sexually mature Patagonian pejerrey O. hatcheri, which have been partially depleted of endogenous GCs by a combination of Busulfan (40 mg/kg and high water temperature (25 degrees C treatments. The observation of the donor cells' behavior showed that transplanted spermatogonial cells were able to recolonize the recipients' gonads and resume spermatogenesis within 6 months from the GCT. The presence of donor-derived gametes was confirmed by PCR in 20% of the surrogate O. hatcheri fathers at 6 months and crosses with O. bonariensis mothers produced hybrids and pure O. bonariensis, with donor-derived germline transmission rates of 1.2-13.3%. These findings indicate that transplantation of spermatogonial cells into sexually competent fish can shorten considerably the production time of donor-derived gametes and offspring and could play a vital role in germline conservation and propagation of valued and/or endangered fish species.

  11. Corona cell RNA sequencing from individual oocytes revealed transcripts and pathways linked to euploid oocyte competence and live birth.

    Science.gov (United States)

    Parks, Jason C; Patton, Alyssa L; McCallie, Blair R; Griffin, Darren K; Schoolcraft, William B; Katz-Jaffe, Mandy G

    2016-05-01

    Corona cells surround the oocyte and maintain a close relationship through transzonal processes and gap junctions, and may be used to assess oocyte competence. In this study, the corona cell transcriptome of individual cumulus oocyte complexes (COCs) was investigated. Isolated corona cells were collected from COCs that developed into euploid blastocysts and were transferred in a subsequent frozen embryo transfer. Ten corona cell samples underwent RNA-sequencing to generate unique gene expression profiles. Live birth was compared with negative implantation after the transfer of a euploid blastocyst using bioinformatics and statistical analysis. Individual corona cell samples produced a mean of 21.2 million sequence reads, and 307 differentially expressed transcrpits (P corona cell transcriptome was successfully generated using RNA-sequencing. Key genes and signalling pathways were identified in association with implantation outcome after the transfer of a euploid blastocyst in a frozen embryo transfer. These data could provide novel biomarkers for the non-invasive assessment of embryo viability.

  12. Application of mesenchymal stem cells as a vehicle to deliver replication-competent adenovirus for treating malignant glioma

    Institute of Scientific and Technical Information of China (English)

    Cui Hai; Yong-Min Jin; Wen-Biao Jin; Zhe-Zhu Han; Mei-Nv Cui; Xue-Zhe Piao; Xiong-Hu Shen; Song-Nan Zhang; Hong-Hua Sun

    2012-01-01

    Although gene therapy was regarded as a promising approach for glioma treatment,its therapeutic efficacy was often disappointing because of the lack of efficient drug delivery systems.Mesenchymal stem cells (MSCs) have been reported to have a tropism for brain tumors and thus could be used as delivery vehicles for glioma therapy.Therefore,in this study,we attempted to treat glioma by using MSCs as a vehicle for delivering replication-competent adenovirus.We firstly compared the infectivity of type 3,type 5,and type 35 fiber-modified adenoviruses in MSCs.We also determined suitable adenovirus titer in vitro and then used this titer to analyze the ability of MSCs to deliver replication-competent adenovirus into glioma in vivo.Our results indicated that type 35 fiber-modified adenovirus showed higher infectivity than did naked type 3 or type 5 fiber-modified adenovirus.MSCs carrying replication-competent adenovirus significantly inhibited tumor growth in vivo compared with other control groups.In conclusion,MSCs are an effective vehicle that can successfully transport replication-competent adenovirus into glioma,making it a potential therapeutic strategy for treating malignant glioma.

  13. Method for Preparation and Electroporation of S. aureus and S. epidermidis.

    Science.gov (United States)

    Grosser, Melinda R; Richardson, Anthony R

    2016-01-01

    For bacterial species that are not known to be naturally competent, such as Staphylococcus aureus and Staphylococcus epidermidis, electroporation is an efficient method for introducing genetic material into the cell. The technique utilizes electrical pulses to transiently permeabilize bacterial cell membranes, which allows for the passage of plasmid DNA across the membranes. Here, we describe methods for preparing electrocompetent S. aureus and S. epidermidis cells and outline a procedure for electroporation of the prepared competent cells.

  14. Evaluation of genotoxicity using automated detection of γH2AX in metabolically competent HepaRG cells.

    Science.gov (United States)

    Quesnot, Nicolas; Rondel, Karine; Audebert, Marc; Martinais, Sophie; Glaise, Denise; Morel, Fabrice; Loyer, Pascal; Robin, Marie-Anne

    2016-01-01

    The in situ detection of γH2AX was recently reported to be a promising biomarker of genotoxicity. In addition, the human HepaRG hepatoma cells appear to be relevant for investigating hepatic genotoxicity since they express most of drug metabolizing enzymes and a wild type p53. The aim of this study was to determine whether the automated in situ detection of γH2AX positive HepaRG cells could be relevant for evaluation of genotoxicity after single or long-term repeated in vitro exposure compared to micronucleus assay. Metabolically competent HepaRG cells were treated daily with environmental contaminants and genotoxicity was evaluated after 1, 7 and 14 days. Using these cells, we confirmed the genotoxicity of aflatoxin B1 and benzo(a)pyrene and demonstrated that dimethylbenzanthracene, fipronil and endosulfan previously found genotoxic with comet or micronucleus assays also induced γH2AX phosphorylation. Furthermore, we showed that fluoranthene and bisphenol A induced γH2AX while no effect had been previously reported in HepG2 cells. In addition, induction of γH2AX was observed with some compounds only after 7 days, highlighting the importance of studying long-term effects of low doses of contaminants. Together, our data demonstrate that automated γH2AX detection in metabolically competent HepaRG cells is a suitable high-through put genotoxicity screening assay.

  15. Periodic retinoic acid-STRA8 signaling intersects with periodic germ-cell competencies to regulate spermatogenesis.

    Science.gov (United States)

    Endo, Tsutomu; Romer, Katherine A; Anderson, Ericka L; Baltus, Andrew E; de Rooij, Dirk G; Page, David C

    2015-05-05

    Mammalian spermatogenesis--the transformation of stem cells into millions of haploid spermatozoa--is elaborately organized in time and space. We explored the underlying regulatory mechanisms by genetically and chemically perturbing spermatogenesis in vivo, focusing on spermatogonial differentiation, which begins a series of amplifying divisions, and meiotic initiation, which ends these divisions. We first found that, in mice lacking the retinoic acid (RA) target gene Stimulated by retinoic acid gene 8 (Stra8), undifferentiated spermatogonia accumulated in unusually high numbers as early as 10 d after birth, whereas differentiating spermatogonia were depleted. We thus conclude that Stra8, previously shown to be required for meiotic initiation, also promotes (but is not strictly required for) spermatogonial differentiation. Second, we found that injection of RA into wild-type adult males induced, independently, precocious spermatogonial differentiation and precocious meiotic initiation; thus, RA acts instructively on germ cells at both transitions. Third, the competencies of germ cells to undergo spermatogonial differentiation or meiotic initiation in response to RA were found to be distinct, periodic, and limited to particular seminiferous stages. Competencies for both transitions begin while RA levels are low, so that the germ cells respond as soon as RA levels rise. Together with other findings, our results demonstrate that periodic RA-STRA8 signaling intersects with periodic germ-cell competencies to regulate two distinct, cell-type-specific responses: spermatogonial differentiation and meiotic initiation. This simple mechanism, with one signal both starting and ending the amplifying divisions, contributes to the prodigious output of spermatozoa and to the elaborate organization of spermatogenesis.

  16. Reduced interleukin-2 responsiveness impairs the ability of Treg cells to compete for IL-2 in nonobese diabetic mice.

    Science.gov (United States)

    James, Cini R; Buckle, Irina; Muscate, Franziska; Otsuka, Masayuki; Nakao, Mari; Oon, Jack Sh; Steptoe, Raymond J; Thomas, Ranjeny; Hamilton-Williams, Emma E

    2016-05-01

    Enhancement of regulatory T cell (Treg cell) frequency and function is the goal of many therapeutic strategies aimed at treating type 1 diabetes (T1D). The interleukin-2 (IL-2) pathway, which has been strongly implicated in T1D susceptibility in both humans and mice, is a master regulator of Treg cell homeostasis and function. We investigated how IL-2 pathway defects impact Treg cells in T1D-susceptible nonobese diabetic (NOD) mice in comparison with protected C57BL/6 and NOD congenic mice. NOD Treg cells were reduced in frequency specifically in the lymph nodes and expressed lower levels of CD25 and CD39/CD73 immunosuppressive molecules. In the spleen and blood, Treg cell frequency was preserved through expansion of CD25(low), effector phenotype Treg cells. Reduced CD25 expression led to decreased IL-2 signaling in NOD Treg cells. In vivo, treatment with IL-2-anti-IL-2 antibody complexes led to effective upregulation of suppressive molecules on NOD Treg cells in the spleen and blood, but had reduced efficacy on lymph node Treg cells. In contrast, NOD CD8(+) and CD4(+) effector T cells were not impaired in their response to IL-2 therapy. We conclude that NOD Treg cells have an impaired responsiveness to IL-2 that reduces their ability to compete for a limited supply of IL-2.

  17. Sustained ERK Activation Underlies Reprogramming in Regeneration-Competent Salamander Cells and Distinguishes Them from Their Mammalian Counterparts

    Directory of Open Access Journals (Sweden)

    Maximina H. Yun

    2014-07-01

    Full Text Available In regeneration-competent vertebrates, such as salamanders, regeneration depends on the ability of various differentiated adult cell types to undergo natural reprogramming. This ability is rarely observed in regeneration-incompetent species such as mammals, providing an explanation for their poor regenerative potential. To date, little is known about the molecular mechanisms mediating natural reprogramming during regeneration. Here, we have identified the extent of extracellular signal-regulated kinase (ERK activation as a key component of such mechanisms. We show that sustained ERK activation following serum induction is required for re-entry into the cell cycle of postmitotic salamander muscle cells, partially by promoting the downregulation of p53 activity. Moreover, ERK activation induces epigenetic modifications and downregulation of muscle-specific genes such as Sox6. Remarkably, while long-term ERK activation is found in salamander myotubes, only transient activation is seen in their mammalian counterparts, suggesting that the extent of ERK activation could underlie differences in regenerative competence between species.

  18. Sustained ERK activation underlies reprogramming in regeneration-competent salamander cells and distinguishes them from their mammalian counterparts.

    Science.gov (United States)

    Yun, Maximina H; Gates, Phillip B; Brockes, Jeremy P

    2014-07-08

    In regeneration-competent vertebrates, such as salamanders, regeneration depends on the ability of various differentiated adult cell types to undergo natural reprogramming. This ability is rarely observed in regeneration-incompetent species such as mammals, providing an explanation for their poor regenerative potential. To date, little is known about the molecular mechanisms mediating natural reprogramming during regeneration. Here, we have identified the extent of extracellular signal-regulated kinase (ERK) activation as a key component of such mechanisms. We show that sustained ERK activation following serum induction is required for re-entry into the cell cycle of postmitotic salamander muscle cells, partially by promoting the downregulation of p53 activity. Moreover, ERK activation induces epigenetic modifications and downregulation of muscle-specific genes such as Sox6. Remarkably, while long-term ERK activation is found in salamander myotubes, only transient activation is seen in their mammalian counterparts, suggesting that the extent of ERK activation could underlie differences in regenerative competence between species.

  19. Proliferation of latently infected CD4+ T cells carrying replication-competent HIV-1: Potential role in latent reservoir dynamics

    Science.gov (United States)

    Hosmane, Nina N.; Kwon, Kyungyoon J.; Bruner, Katherine M.; Capoferri, Adam A.; Rosenbloom, Daniel I.S.; Keele, Brandon F.; Ho, Ya-Chi

    2017-01-01

    A latent reservoir for HIV-1 in resting CD4+ T lymphocytes precludes cure. Mechanisms underlying reservoir stability are unclear. Recent studies suggest an unexpected degree of infected cell proliferation in vivo. T cell activation drives proliferation but also reverses latency, resulting in productive infection that generally leads to cell death. In this study, we show that latently infected cells can proliferate in response to mitogens without producing virus, generating progeny cells that can release infectious virus. Thus, assays relying on one round of activation underestimate reservoir size. Sequencing of independent clonal isolates of replication-competent virus revealed that 57% had env sequences identical to other isolates from the same patient. Identity was confirmed by full-genome sequencing and was not attributable to limited viral diversity. Phylogenetic and statistical analysis suggested that identical sequences arose from in vivo proliferation of infected cells, rather than infection of multiple cells by a dominant viral species. The possibility that much of the reservoir arises by cell proliferation presents challenges to cure. PMID:28341641

  20. Solid oxide cell stack and method for preparing same

    DEFF Research Database (Denmark)

    2012-01-01

    A method for producing and reactivating a solid oxide cell stack structure by providing a catalyst precursor in at least one of the electrode layers by impregnation and subsequent drying after the stack has been assembled and initiated. Due to a significantly improved performance and an unexpected...... voltage improvement this solid oxide cell stack structure is particularly suitable for use in solid oxide fuel cell (SOFC) and solid oxide electrolysing cell (SOEC) applications....

  1. Preparing normal tissue cells for space flight experiments.

    Science.gov (United States)

    Koch, Claudia; Kohn, Florian P M; Bauer, Johann

    2016-01-01

    Deterioration of health is a problem in modern space flight business. In order to develop countermeasures, research has been done on human bodies and also on single cells. Relevant experiments on human cells in vitro are feasible when microgravity is simulated by devices such as the Random Positioning Machine or generated for a short time during parabolic flights. However, they become difficult in regard to performance and interpretation when long-term experiments are designed that need a prolonged stay on the International Space Station (ISS). One huge problem is the transport of living cells from a laboratory on Earth to the ISS. For this reason, mainly rapidly growing, rather robust human cells such as cancer cells, embryonic cells, or progenitor cells have been investigated on the ISS up to now. Moreover, better knowledge on the behavior of normal mature cells, which mimic the in vivo situation, is strongly desirable. One solution to the problem could be the use of redifferentiable cells, which grow rapidly and behave like cancer cells in plain medium, but are reprogrammed to normal cells when substances like retinoic acid are added. A list of cells capable of redifferentiation is provided, together with names of suitable drugs, in this review.

  2. Utilization of replication-competent XMRV reporter-viruses reveals severe viral restriction in primary human cells.

    Directory of Open Access Journals (Sweden)

    Christina Martina Stürzel

    Full Text Available The gammaretrovirus termed xenotropic murine leukemia virus-related virus (XMRV was described to be isolated from prostate cancer tissue biopsies and from blood of patients suffering from chronic fatigue syndrome. However, many studies failed to detect XMRV and to verify these disease associations. Data suggesting the contamination of specimens in particular by PCR-based methods and recent reports demonstrating XMRV generation via recombination of two murine leukemia virus precursors raised serious doubts about XMRV being a genuine human pathogen. To elucidate cell tropism of XMRV, we generated replication competent XMRV reporter viruses encoding a green fluorescent protein or a secretable luciferase as tools to analyze virus infection of human cell lines or primary human cells. Transfection of proviral DNAs into LNCaP prostate cancer cells resulted in readily detectably reporter gene expression and production of progeny virus. Inoculation of known XMRV susceptible target cells revealed that these virions were infectious and expressed the reporter gene, allowing for a fast and highly sensitive quantification of XMRV infection. Both reporter viruses were capable of establishing a spreading infection in LNCaP and Raji B cells and could be easily passaged. However, after inoculation of primary human blood cells such as CD4 T cells, macrophages or dendritic cells, infection rates were very low, and a spreading infection was never established. In line with these results we found that supernatants derived from these XMRV infected primary cell types did not contain infectious virus. Thus, although XMRV efficiently replicated in some human cell lines, all tested primary cells were largely refractory to XMRV infection and did not support viral spread. Our results provide further evidence that XMRV is not a human pathogen.

  3. Mathematical Competences

    DEFF Research Database (Denmark)

    Westphael, Henning; Mogensen, Arne

    2013-01-01

    In this article we present the notion of Mathematical competences as a tool to describe the mathematically gifted students.......In this article we present the notion of Mathematical competences as a tool to describe the mathematically gifted students....

  4. A cell-block preparation using glucomannan extracted from Amorphophallus konjac.

    Science.gov (United States)

    Kaneko, Chiyuki; Kobayashi, Tadao K; Hasegawa, Kiyoshi; Udagawa, Yasuhiro; Iwai, Muneo

    2010-09-01

    To evaluate a cell-block preparation using glucomannan, which was extracted from Amorphophallus konjac. Ten specimens were centrifuged at 1,500 rpm for 5 minutes, the supernatant was removed; the remnant after the preparation of smear specimens for routine cytological examination was fixed with 20% formalin. The specimen was recentrifuged at 1,500 rpm for 5 minutes, and the supernatant was removed. The residue was resuspended with 2 ml of eosin solution and 1-5 ml of 80% alcohol, and stirred well. After further centrifugation, the supernatant was removed, and one drop of a glucomannan-formalin water solution was added gently. After immersion in methanol for 2 hours, glucomannan is solidified and becomes gelatinous. The obtained cell block was placed in the cassette for the preparation of tissue specimens, dehydrated by the routine method, infiltrated with paraffin, and a paraffin-embedded block was prepared. Thin sections were prepared from the paraffin-embedded cell block, and hematoxylin-eosin (H&E) stain with immunological stains was performed. H&E stain, periodic acid-Schiff reaction, Alcian blue, and immunohistochemical stain were clearly demonstrated.We evaluated a new modality of cell-block preparation using a glucomannan-formalin water solution. We found that the method was easy to perform and thought it could be useful as an alternative technique for cell-block preparations. Thus, this novel technique should find wide application in the future. Copyright 2009 Wiley-Liss, Inc.

  5. Reduced Intensity Preparative Regimen Followed by Stem Cell Transplant (FAB)

    Science.gov (United States)

    2016-03-29

    Myelodysplastic and Myeloproliferative Disorders; Acute Myelogenous Leukemia; Acute Lymphoblastic Leukemia; Chronic Myelogenous Leukemia; Multiple Myeloma; Plasma Cell Dyscrasia; Lymphoproliferative Disorders; Hematologic Diseases

  6. Preparation of Drosophila S2 cells for Light Microscopy

    Science.gov (United States)

    Buster, Daniel W.; Nye, Jonathan; Klebba, Joseph E.; Rogers, Gregory C.

    2010-01-01

    The ideal experimental system would be cheap and easy to maintain, amenable to a variety of techniques, and would be supported by an extensive literature and genome sequence database. Cultured Drosophila S2 cells, the product of disassociated 20-24 hour old embryos1, possess all these properties. Consequently, S2 cells are extremely well-suited for the analysis of cellular processes, including the discovery of the genes encoding the molecular components of the process or mechanism of interest. The features of S2 cells that are most responsible for their utility are the ease with which they are maintained, their exquisite sensitivity to double-stranded (ds)RNA-mediated interference (RNAi), and their tractability to fluorescence microscopy as either live or fixed cells. S2 cells can be grown in a variety of media, including a number of inexpensive, commercially-available, fully-defined, serum-free media2. In addition, they grow optimally and quickly at 21-24°C and can be cultured in a variety of containers. Unlike mammalian cells, S2 cells do not require a regulated atmosphere, but instead do well with normal air and can even be maintained in sealed flasks. Complementing the ease of RNAi in S2 cells is the ability to readily analyze experimentally-induced phenotypes by phase or fluorescence microscopy of fixed or live cells. S2 cells grow in culture as a single monolayer but do not display contact inhibition. Instead, cells tend to grow in colonies in dense cultures. At low density, S2 cultures grown on glass or tissue culture-treated plastic are round and loosely-attached. However, the cytology of S2 cells can be greatly improved by inducing them to flatten extensively by briefly culturing them on a surface coated with the lectin, concanavalin A (ConA)3. S2 cells can also be stably transfected with fluorescently-tagged markers to label structures or organelles of interest in live or fixed cells. Therefore, the usual scenario for the microscopic analysis of cells is

  7. Fuel cell membrane preparation: effects of base polymer

    Energy Technology Data Exchange (ETDEWEB)

    Brack, H.P.; Scherer, G.G. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1997-06-01

    Radiation grafted films and membranes prepared from the partially fluorinated base copolymer poly(ethylene-alt-tetrafluoroethylene) or ETFE have better mechanical properties than those prepared from poly(tetrafluoroethylene-co-hexafluoropropylene) or FEP. The influence of the base copolymer film type on the grafting rate and yields is reported in the present investigation. An understanding of the effects of these parameters is important so that the grafting process can be carried out reproducibly in as short a time as possible. The grafting rate and yield as a function of the irradiation dose has been found to be much higher for the partially fluorinated base copolymer ETFE. (author) 2 figs., 1 tab., 5 refs.

  8. Model for solid oxide fuel cell cathodes prepared by infiltration

    DEFF Research Database (Denmark)

    Samson, Alfred Junio; Søgaard, Martin; Hendriksen, Peter Vang

    2017-01-01

    A 1-dimensional model of a cathode has been developed in order to understand and predict the performance of cathodes prepared by infiltration of La0.6Sr0.4Co1.05O3-δ (LSC) into porous backbones of Ce0.9Gd0.1O1.95 (CGO). The model accounts for the mixed ionic and electronic conductivity of LSC......, ionic conductivity of CGO, gas transport in the porous cathode, and the oxygen reduction reaction at the surface of percolated LSC. Geometrical variations are applied to reflect a changing microstructure of LSC under varying firing temperatures. Using microstructural parameters obtained from detailed...... parameter variations are presented and discussed with the aim of presenting specific guidelines for optimizing the microstructure of cathodes prepared by infiltration....

  9. Forming key competences from the position of competence approach

    OpenAIRE

    Latypova E. R.

    2012-01-01

    Teaching foreign languages pursues practical, educational, bringing-up and developing aims in non-linguistic educational institution. The practical aim in teaching foreign languages is preparation of a highly trained specialist, which masters key competences and can work hard on his specialty according to world standards and is ready to act in professional communication with foreigners. New requirements direct the system of education toward competence approach. The main aim of competence appr...

  10. The small GTPase Cdc42 modulates the number of exocytosis-competent dense-core vesicles in PC12 cells

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Mai [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902 (Japan); Kitaguchi, Tetsuya [Cell Signaling Group, Waseda Bioscience Research Institute in Singapore (WABOIS), Waseda University, 11 Biopolis Way, 05-01/02 Helios, Singapore 138667 (Singapore); Numano, Rika [The Electronics-Inspired Interdisciplinary Research Institute (EIIRIS), Toyohashi University of Technology, 1-1 Hibarigaoka, Tennpaku-cho, Toyohashi, Aichi 441-8580 (Japan); Ikematsu, Kazuya [Forensic Pathology and Science, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8523 (Japan); Kakeyama, Masaki [Laboratory of Environmental Health Sciences, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033 (Japan); Murata, Masayuki; Sato, Ken [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902 (Japan); Tsuboi, Takashi, E-mail: takatsuboi@bio.c.u-tokyo.ac.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902 (Japan)

    2012-04-06

    Highlights: Black-Right-Pointing-Pointer Regulation of exocytosis by Rho GTPase Cdc42. Black-Right-Pointing-Pointer Cdc42 increases the number of fusion events from newly recruited vesicles. Black-Right-Pointing-Pointer Cdc42 increases the number of exocytosis-competent dense-core vesicles. -- Abstract: Although the small GTPase Rho family Cdc42 has been shown to facilitate exocytosis through increasing the amount of hormones released, the precise mechanisms regulating the quantity of hormones released on exocytosis are not well understood. Here we show by live cell imaging analysis under TIRF microscope and immunocytochemical analysis under confocal microscope that Cdc42 modulated the number of fusion events and the number of dense-core vesicles produced in the cells. Overexpression of a wild-type or constitutively-active form of Cdc42 strongly facilitated high-KCl-induced exocytosis from the newly recruited plasma membrane vesicles in PC12 cells. By contrast, a dominant-negative form of Cdc42 inhibited exocytosis from both the newly recruited and previously docked plasma membrane vesicles. The number of intracellular dense-core vesicles was increased by the overexpression of both a wild-type and constitutively-active form of Cdc42. Consistently, activation of Cdc42 by overexpression of Tuba, a Golgi-associated guanine nucleotide exchange factor for Cdc42 increased the number of intracellular dense-core vesicles, whereas inhibition of Cdc42 by overexpression of the Cdc42/Rac interactive binding domain of neuronal Wiskott-Aldrich syndrome protein decreased the number of them. These findings suggest that Cdc42 facilitates exocytosis by modulating both the number of exocytosis-competent dense-core vesicles and the production of dense-core vesicles in PC12 cells.

  11. Sample preparation by cell guiding using negative dielectrophoresis

    DEFF Research Database (Denmark)

    Christensen, Troels Balmer; Pedersen, Christian Møller; Bang, Dang Duong

    2007-01-01

    and a decreased efficiency with increased flowrate is observed. In addition, the DEP behaviour of the yeast cells at different medium conductivities and applied frequencies is investigated. The chip is easily fabricated in a two-step process: Standard UV lithography techniques are used for electrode fabrication......In this study, we present a microsystem designed for performing and testing dielectrophoretic (DEP) guiding of biological cells. Baker's yeast (Saccharomyces cerevisiae) is used as a model organism to study cell guiding in the system. The guiding efficiency as a function of flowrate is investigated...

  12. Where Cultural Competency Begins: Changes in Undergraduate Students' Intercultural Competency

    Science.gov (United States)

    Sandell, Elizabeth J.; Tupy, Samantha J.

    2015-01-01

    Teacher preparation programs and accreditation organizations have acknowledged need for educators to demonstrate intercultural knowledge, skills, and abilities. Teacher educators are responding to emphasis in higher education to assure that graduates achieve intercultural competence (NCATE, 2008). This study compared the cultural competency of…

  13. A sealed preparation for long-term observations of cultured cells.

    Science.gov (United States)

    Sluder, Greenfield; Nordberg, Joshua J; Miller, Frederick J; Hinchcliffe, Edward H

    2007-01-01

    INTRODUCTIONThe continuous long-term observation of cultured cells on the microscope has always been a technically demanding undertaking. This protocol describes a sealed preparation that allows the continuous long-term observation of cultured mammalian cells on upright or inverted microscopes without environmental CO(2) control. The preparation allows for optical conditions consistent with high-quality imaging and good cell viability for at least 100 hours. The preparation is an aluminum support slide with a square aperture cut in its center. The coverslip bearing the cells is attached to the top of the slide with a thin layer of silicone grease, and the bottom of the slide is similarly covered with a clean coverslip of the same size. The thickness of the slide is intended to coordinately maximize the volume of the medium while maintaining optical properties that allow Koehler illumination with standard condensers. The chamber is filled in equal parts with HEPES-buffered media containing fetal calf serum and a low-viscosity fluorocarbon oil. These oils have a high solubility for atmospheric gases. The inclusion of the oil in the preparations is intended to provide a source of oxygen and perhaps a sink for some of the CO(2) produced by the cells. Although the inclusion of fluorocarbon oil in the preparation may not be necessary for short-term (~24 hr) observations, particularly with cells that are sparsely plated, long-term cell viability is ensured when the oil is present.

  14. Multiparameter Analysis of Human Bone Marrow Stromal Cells Identifies Distinct Immunomodulatory and Differentiation-Competent Subtypes

    NARCIS (Netherlands)

    S. James (Sally); J. Fox (James); F. Afsari (Farinaz); J. Lee (Jennifer); S. Clough (Sally); C. Knight (Charlotte); J. Ashmore (James); P. Ashton (Peter); O. Preham (Olivier); M.J. Hoogduijn (Martin); R.D.A.R. Ponzoni (Raquel De Almeida Rocha); Y. Hancock; M. Coles (Mark); P.G. Genever (Paul)

    2015-01-01

    textabstractBone marrow stromal cells (BMSCs, also called bone-marrow-derived mesenchymal stromal cells) provide hematopoietic support and immunoregulation and contain a stem cell fraction capable of skeletogenic differentiation. We used immortalized human BMSC clonal lines for multi-level analysis

  15. Competencies: requirements and acquisition

    NARCIS (Netherlands)

    Kuenn, A.C.; Meng, C.M.; Peters, Z.; Verhagen, A.M.C.

    2013-01-01

    Higher education is given the key task to prepare the highly talented among the young to fulfil highly qualified roles in the labour market. Successful labour market performance of graduates is generally associated with the acquisition of the correct competencies. Education as an individual investme

  16. Developing Soldier Cultural Competency

    Science.gov (United States)

    2006-03-03

    providing Soldiers a “ tourist ” level of understanding which does not adequately prepare them to be culturally competent and effective. The one exception is...cultures use high-context, indirect communications. High- 7 context groups view the low context groups as abrupt and rude in their messages, while low

  17. MicroRNA and protein profiling of brain metastasis competent cell-derived exosomes.

    Directory of Open Access Journals (Sweden)

    Laura Camacho

    Full Text Available Exosomes are small membrane vesicles released by most cell types including tumor cells. The intercellular exchange of proteins and genetic material via exosomes is a potentially effective approach for cell-to-cell communication and it may perform multiple functions aiding to tumor survival and metastasis. We investigated microRNA and protein profiles of brain metastatic (BM versus non-brain metastatic (non-BM cell-derived exosomes. We studied the cargo of exosomes isolated from brain-tropic 70W, MDA-MB-231BR, and circulating tumor cell brain metastasis-selected markers (CTC1BMSM variants, and compared them with parental non-BM MeWo, MDA-MB-231P and CTC1P cells, respectively. By performing microRNA PCR array we identified one up-regulated (miR-210 and two down-regulated miRNAs (miR-19a and miR-29c in BM versus non-BM exosomes. Second, we analyzed the proteomic content of cells and exosomes isolated from these six cell lines, and detected high expression of proteins implicated in cell communication, cell cycle, and in key cancer invasion and metastasis pathways. Third, we show that BM cell-derived exosomes can be internalized by non-BM cells and that they effectively transport their cargo into cells, resulting in increased cell adhesive and invasive potencies. These results provide a strong rationale for additional investigations of exosomal proteins and miRNAs towards more profound understandings of exosome roles in brain metastasis biogenesis, and for the discovery and application of non-invasive biomarkers for new therapies combating brain metastasis.

  18. MicroRNA and protein profiling of brain metastasis competent cell-derived exosomes.

    Science.gov (United States)

    Camacho, Laura; Guerrero, Paola; Marchetti, Dario

    2013-01-01

    Exosomes are small membrane vesicles released by most cell types including tumor cells. The intercellular exchange of proteins and genetic material via exosomes is a potentially effective approach for cell-to-cell communication and it may perform multiple functions aiding to tumor survival and metastasis. We investigated microRNA and protein profiles of brain metastatic (BM) versus non-brain metastatic (non-BM) cell-derived exosomes. We studied the cargo of exosomes isolated from brain-tropic 70W, MDA-MB-231BR, and circulating tumor cell brain metastasis-selected markers (CTC1BMSM) variants, and compared them with parental non-BM MeWo, MDA-MB-231P and CTC1P cells, respectively. By performing microRNA PCR array we identified one up-regulated (miR-210) and two down-regulated miRNAs (miR-19a and miR-29c) in BM versus non-BM exosomes. Second, we analyzed the proteomic content of cells and exosomes isolated from these six cell lines, and detected high expression of proteins implicated in cell communication, cell cycle, and in key cancer invasion and metastasis pathways. Third, we show that BM cell-derived exosomes can be internalized by non-BM cells and that they effectively transport their cargo into cells, resulting in increased cell adhesive and invasive potencies. These results provide a strong rationale for additional investigations of exosomal proteins and miRNAs towards more profound understandings of exosome roles in brain metastasis biogenesis, and for the discovery and application of non-invasive biomarkers for new therapies combating brain metastasis.

  19. PREPARATION AND THE CULTURE OF LO2 CELLS ON PVA-BASED MICROCARRIERS

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hong; YU Yaoting; PAN Jilun; WANG lianyong

    2001-01-01

    Using polyvinyl alcohol (PVA) as raw material and vacuum pump oil as oil phase medium,PVA-based microcarriers were prepared by suspension method The diameters of the beads were 100-180 μ m. LO2 cells were cultured on PVA-based microcarriers and cytodexⅢ microcarriers.Morphology, attachment and growth rate of LO2 cells were studied.

  20. PREPARATION AND THE CULTURE OF LO2 CELLS ON PVA—BASED MICROCARRIERS

    Institute of Scientific and Technical Information of China (English)

    ZHANGHong; YUYaoting; 等

    2001-01-01

    Using polyvinyl alcohol(PVA) as raw material and vacuum pump oil as oil phase medium,PVA-based microcarriers were prepared by suspension method,The diameters of the beads were 100-180um,LO2 cells were cultured on PVA-based microcarriers and cytodexⅢ microcarriers.Morphology,attachment and growth rate of LO2 cells were studied.

  1. Immune-Pineal Axis: Nuclear Factor κB (NF-kB Mediates the Shift in the Melatonin Source from Pinealocytes to Immune Competent Cells

    Directory of Open Access Journals (Sweden)

    Regina P. Markus

    2013-05-01

    Full Text Available Pineal gland melatonin is the darkness hormone, while extra-pineal melatonin produced by the gonads, gut, retina, and immune competent cells acts as a paracrine or autocrine mediator. The well-known immunomodulatory effect of melatonin is observed either as an endocrine, a paracrine or an autocrine response. In mammals, nuclear translocation of nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB blocks noradrenaline-induced melatonin synthesis in pinealocytes, which induces melatonin synthesis in macrophages. In addition, melatonin reduces NF-κB activation in pinealocytes and immune competent cells. Therefore, pathogen- or danger-associated molecular patterns transiently switch the synthesis of melatonin from pinealocytes to immune competent cells, and as the response progresses melatonin inhibition of NF-κB activity leads these cells to a more quiescent state. The opposite effect of NF-κB in pinealocytes and immune competent cells is due to different NF-κB dimers recruited in each phase of the defense response. This coordinated shift of the source of melatonin driven by NF-κB is called the immune-pineal axis. Finally, we discuss how this concept might be relevant to a better understanding of pathological conditions with impaired melatonin rhythms and hope it opens new horizons for the research of side effects of melatonin-based therapies.

  2. Preparation of crude rough microsomes from tissue culture cells.

    Science.gov (United States)

    Sabatini, David D

    2014-09-02

    There are various procedures for isolating microsomal fractions from tissue culture cells. The essential conditions for each step of one procedure are described here. Notes for special circumstances are included so that the procedure can be modified according to the experimental purpose.

  3. Preparation and characterization of polymeric membranes for fuel cells

    KAUST Repository

    Nunes, Suzana Pereira

    2012-06-24

    This paper summarized part of the material development for fuel cell conducted in my group in the last 10 years, based on new functionalized polymers with phosphonic, sulfonic as well as oxadiazole and triazole sites. © 2012 Bentham Science Publishers. All rights reserved.

  4. IL-17 regulates systemic fungal immunity by controlling the functional competence of NK cells.

    Science.gov (United States)

    Bär, Eva; Whitney, Paul G; Moor, Kathrin; Reis e Sousa, Caetano; LeibundGut-Landmann, Salomé

    2014-01-16

    Interleukin 17 (IL-17)-mediated immunity plays a key role in protection from fungal infections in mice and man. Here, we confirmed that mice deficient in the IL-17 receptor or lacking the ability to secrete IL-17 are highly susceptible to systemic candidiasis, but we found that temporary blockade of the IL-17 pathway during infection in wild-type mice did not impact fungal control. Rather, mice lacking IL-17 receptor signaling had a cell-intrinsic impairment in the development of functional NK cells, which accounted for the susceptibility of these mice to systemic fungal infection. NK cells promoted antifungal immunity by secreting GM-CSF, necessary for the fungicidal activity of neutrophils. These data reveal that NK cells are crucial for antifungal defense and indicate a role for IL-17 family cytokines in NK cell development. The IL-17-NK cell axis may impact immunity against not only fungi but also bacteria, viruses, and tumors.

  5. Preparation of Epidermal Peels and Guard Cell Protoplasts for Cellular, Electrophysiological, and -Omics Assays of Guard Cell Function.

    Science.gov (United States)

    Zhu, Mengmeng; Jeon, Byeong Wook; Geng, Sisi; Yu, Yunqing; Balmant, Kelly; Chen, Sixue; Assmann, Sarah M

    2016-01-01

    Bioassays are commonly used to study stomatal phenotypes. There are multiple options in the choice of plant materials and species used for observation of stomatal and guard cell responses in vivo. Here, detailed procedures for bioassays of stomatal responses to abscisic acid (ABA) in Arabidopsis thaliana are described, including ABA promotion of stomatal closure, ABA inhibition of stomatal opening, and ABA promotion of reaction oxygen species (ROS) production in guard cells. We also include an example of a stomatal bioassay for the guard cell CO2 response using guard cell-enriched epidermal peels from Brassica napus. Highly pure preparations of guard cell protoplasts can be produced, which are also suitable for studies on guard cell signaling, as well as for studies on guard cell ion transport. Small-scale and large-scale guard cell protoplast preparations are commonly used for electrophysiological and -omics studies, respectively. We provide a procedure for small-scale guard cell protoplasting from A. thaliana. Additionally, a general protocol for large-scale preparation of guard cell protoplasts, with specifications for three different species, A. thaliana, B. napus, and Vicia faba is also provided.

  6. Establishment of Germ-line Competent C57BL/6J Embryonic Stem Cell Lines

    Institute of Scientific and Technical Information of China (English)

    Gui-jun YAN; Zheng GU; Jian WANG; Jia-ke TSO

    2004-01-01

    Objective To establish C57BL/6J embryonic stem (ES) cell lines with potential germline contribution Methods ES cells were isolated from blastocyst inner cell mass of C57BL/6J mice, and cultured for 15 passages, and then injected into blastococels of lCR mice blastocysts to establish chimeric mice.Results Three ES cell lines (mC57ESl,mC57ES3, mC57ES7) derived from the inner cell mass of C57BL/6J mice blastocysts were established. They were characteristic of undifferentiated state, including normal XY karyotype, expression of a specific cell surface marker "stage-specific embryonic antigen-1" and alkaline phosphatase in continuous passage. When injected into immunodeficient mice, mC5 7ES1 cells consis tently differentiated into derivatives of all three embryonic germ layers. When mC57ES1cells were transferred into ICR mice blastocysts, 4 chimeric mice have been obtained.One male of them revealed successful germ-line transmission. Conclussion We have obtained C57BL/6J ES cell lines with a potential germ-line contribution, which can be used to generate transgenic and gene knock-out mice.

  7. Dorsomorphin promotes survival and germline competence of zebrafish spermatogonial stem cells in culture.

    Directory of Open Access Journals (Sweden)

    Ten-Tsao Wong

    Full Text Available Zebrafish spermatogonial cell cultures were established from Tg(piwil1:neo;Tg(piwil1:DsRed transgenic fish using a zebrafish ovarian feeder cell line (OFC3 that was engineered to express zebrafish Lif, Fgf2 and Gdnf. Primary cultures, initiated from testes, were treated with G418 to eliminate the somatic cells and select for the piwil1:neo expressing spermatogonia. Addition of dorsomorphin, a Bmp type I receptor inhibitor, prolonged spermatogonial stem cell (SSC survival in culture and enhanced germline transmission of the SSCs following transplantation into recipient larvae. In contrast, dorsomorphin inhibited the growth and survival of zebrafish female germline stem cells (FGSCs in culture. In the presence of dorsomorphin, the spermatogonia continued to express the germ-cell markers dazl, dnd, nanos3, vasa and piwil1 and the spermatogonial markers plzf and sox17 for at least six weeks in culture. Transplantation experiments revealed that 6 week-old spermatogonial cell cultures maintained in the presence of dorsomorphin were able to successfully colonize the gonad in 18% of recipient larvae and produce functional gametes in the resulting adult chimeric fish. Germline transmission was not successful when the spermatogonia were cultured 6 weeks in the absence of dorsomorphin before transplantation. The results indicate that Bmp signaling is detrimental to SSCs but required for the survival of zebrafish FGSCs in culture. Manipulation of Bmp signaling could provide a strategy to optimize culture conditions of germline stem cells from other species.

  8. In vivo assay for the developmental competence of embryo-derived zebrafish cell lines

    NARCIS (Netherlands)

    Speksnijder, JE; Hage, WJ; Lanser, PH; Collodi, P; Zivkovic, D

    1997-01-01

    We have produced chimeric zebrafish embryos by transplanting permanent embryo-derived cell lines into blastula-stage embryos. Furthermore, we have established a fluorescent in vivo assay to monitor the developmental effects and fate of such transplanted cells using confocal laser scanning microscopy

  9. Neural-Competent Cells of Adult Human Dermis Belong to the Schwann Lineage

    Science.gov (United States)

    Etxaniz, Usue; Pérez-San Vicente, Adrián; Gago-López, Nuria; García-Dominguez, Mario; Iribar, Haizea; Aduriz, Ariane; Pérez-López, Virginia; Burgoa, Izaskun; Irizar, Haritz; Muñoz-Culla, Maider; Vallejo-Illarramendi, Ainara; Leis, Olatz; Matheu, Ander; Martín, Angel G.; Otaegui, David; López-Mato, María Paz; Gutiérrez-Rivera, Araika; MacLellan, Robb; Izeta, Ander

    2014-01-01

    Summary Resident neural precursor cells (NPCs) have been reported for a number of adult tissues. Understanding their physiological function or, alternatively, their activation after tissue damage or in vitro manipulation remains an unsolved issue. Here, we investigated the source of human dermal NPCs in adult tissue. By following an unbiased, comprehensive approach employing cell-surface marker screening, cell separation, transcriptomic characterization, and in vivo fate analyses, we found that p75NTR+ precursors of human foreskin can be ascribed to the Schwann (CD56+) and perivascular (CD56−) cell lineages. Moreover, neural differentiation potential was restricted to the p75NTR+CD56+ Schwann cells and mediated by SOX2 expression levels. Double-positive NPCs were similarly obtained from human cardiospheres, indicating that this phenomenon might be widespread. PMID:25418723

  10. Neural-Competent Cells of Adult Human Dermis Belong to the Schwann Lineage

    Directory of Open Access Journals (Sweden)

    Usue Etxaniz

    2014-11-01

    Full Text Available Resident neural precursor cells (NPCs have been reported for a number of adult tissues. Understanding their physiological function or, alternatively, their activation after tissue damage or in vitro manipulation remains an unsolved issue. Here, we investigated the source of human dermal NPCs in adult tissue. By following an unbiased, comprehensive approach employing cell-surface marker screening, cell separation, transcriptomic characterization, and in vivo fate analyses, we found that p75NTR+ precursors of human foreskin can be ascribed to the Schwann (CD56+ and perivascular (CD56− cell lineages. Moreover, neural differentiation potential was restricted to the p75NTR+CD56+ Schwann cells and mediated by SOX2 expression levels. Double-positive NPCs were similarly obtained from human cardiospheres, indicating that this phenomenon might be widespread.

  11. Neural-competent cells of adult human dermis belong to the Schwann lineage.

    Science.gov (United States)

    Etxaniz, Usue; Pérez-San Vicente, Adrián; Gago-López, Nuria; García-Dominguez, Mario; Iribar, Haizea; Aduriz, Ariane; Pérez-López, Virginia; Burgoa, Izaskun; Irizar, Haritz; Muñoz-Culla, Maider; Vallejo-Illarramendi, Ainara; Leis, Olatz; Matheu, Ander; Martín, Angel G; Otaegui, David; López-Mato, María Paz; Gutiérrez-Rivera, Araika; MacLellan, Robb; Izeta, Ander

    2014-11-11

    Resident neural precursor cells (NPCs) have been reported for a number of adult tissues. Understanding their physiological function or, alternatively, their activation after tissue damage or in vitro manipulation remains an unsolved issue. Here, we investigated the source of human dermal NPCs in adult tissue. By following an unbiased, comprehensive approach employing cell-surface marker screening, cell separation, transcriptomic characterization, and in vivo fate analyses, we found that p75NTR(+) precursors of human foreskin can be ascribed to the Schwann (CD56(+)) and perivascular (CD56(-)) cell lineages. Moreover, neural differentiation potential was restricted to the p75NTR(+)CD56(+) Schwann cells and mediated by SOX2 expression levels. Double-positive NPCs were similarly obtained from human cardiospheres, indicating that this phenomenon might be widespread.

  12. Cell dynamics in the pulpal healing process following cavity preparation in rat molars.

    Science.gov (United States)

    Harada, Masahiro; Kenmotsu, Shin-Ichi; Nakasone, Naohiro; Nakakura-Ohshima, Kuniko; Ohshima, Hayato

    2008-10-01

    Odontoblast-lineage cells acquire heat-shock protein (HSP)-25-immunoreactivity (IR) after they complete their cell division, suggesting that this protein acts as a switch between cell proliferation and differentiation during tooth development. However, there are few available data concerning the relationship between cell proliferation and differentiation following cavity preparation. The present study aims to clarify the expression of HSP-25 in the odontoblast-lineage cells with their proliferative activity after cavity preparation by immunocytochemistry for HSP-25 and cell proliferation assay using 5-bromo-2'-deoxyuridine (BrdU) labeling. In untreated control teeth, intense HSP-25-IR was found in odontoblasts and some subodontoblastic mesenchymal cells. Cavity preparation caused the destruction of odontoblasts and the disappearance of HSP-25-IR was conspicuous at the affected site, although some cells retained HSP-25-IR and subsequently most of them disappeared from the pulp-dentin border by postoperative day 1. Contrary, some subodontoblastic mesenchymal cells with weak HSP-25-IR began to take the place of degenerated cells, although no proliferative activity was recognizable in the dental pulp. Interestingly, proliferative cells in the dental pulp significantly increased in number on day 2 when the newly differentiating cells already arranged along the pulp-dentin border, and continued their proliferative activity in the wide range of the pulp tissue until day 5. These findings indicate that progenitor cells equipped in the subodontoblastic layer firstly migrate and differentiate into new odontoblast-like cells to compensate for the loss of the odontoblast layer, and subsequently the reorganization of dental pulp was completed by active proliferation of the mesenchymal cells occurring in a wide range of pulp tissue.

  13. Noble metal-free hydrazine fuel cell catalysts: EPOC effect in competing chemical and electrochemical reaction pathways.

    Science.gov (United States)

    Sanabria-Chinchilla, Jean; Asazawa, Koichiro; Sakamoto, Tomokazu; Yamada, Koji; Tanaka, Hirohisa; Strasser, Peter

    2011-04-13

    We report the discovery of a highly active Ni-Co alloy electrocatalyst for the oxidation of hydrazine (N(2)H(4)) and provide evidence for competing electrochemical (faradaic) and chemical (nonfaradaic) reaction pathways. The electrochemical conversion of hydrazine on catalytic surfaces in fuel cells is of great scientific and technological interest, because it offers multiple redox states, complex reaction pathways, and significantly more favorable energy and power densities compared to hydrogen fuel. Structure-reactivity relations of a Ni(60)Co(40) alloy electrocatalyst are presented with a 6-fold increase in catalytic N(2)H(4) oxidation activity over today's benchmark catalysts. We further study the mechanistic pathways of the catalytic N(2)H(4) conversion as function of the applied electrode potential using differentially pumped electrochemical mass spectrometry (DEMS). At positive overpotentials, N(2)H(4) is electrooxidized into nitrogen consuming hydroxide ions, which is the fuel cell-relevant faradaic reaction pathway. In parallel, N(2)H(4) decomposes chemically into molecular nitrogen and hydrogen over a broad range of electrode potentials. The electroless chemical decomposition rate was controlled by the electrode potential, suggesting a rare example of a liquid-phase electrochemical promotion effect of a chemical catalytic reaction ("EPOC"). The coexisting electrocatalytic (faradaic) and heterogeneous catalytic (electroless, nonfaradaic) reaction pathways have important implications for the efficiency of hydrazine fuel cells. © 2011 American Chemical Society

  14. Fish peripheral blood mononuclear cells preparation for future monitoring applications.

    Science.gov (United States)

    Pierrard, Marie-Aline; Roland, Kathleen; Kestemont, Patrick; Dieu, Marc; Raes, Martine; Silvestre, Frédéric

    2012-07-15

    Fish species possess many specific characteristics that support their use in ecotoxicology. Widely used in clinical research, peripheral blood mononuclear cells (PBMCs) can reasonably be exploited as relevant target cells in the assessment of environmental chemical toxicity. The current article focuses on the methods necessary to isolate, characterize, and culture fish PBMCs. These procedures were successfully applied on an endangered species, the European eel (Anguilla anguilla L.), and on an economically important and worldwide exported species, the Asian catfish (Pangasianodon hypophthalmus S.). Proteomic approaches can be useful to screen xenobiotic exposure at the protein expression level, giving the opportunity to develop early warning signals thanks to molecular signatures of toxicity. To date, a major limitation of proteomic analyses is that most protein expression profiles often reveal the same predominant and frequently differentially expressed families of proteins regardless of the experimental stressing conditions. The current study describes a methodology to get a postnuclear fraction of high quality isolated from fish PBMCs in order to perform subsequent subproteomic analyses. Applied on samples from eel, the subproteomic analysis (two-dimensional differential in-gel electrophoresis) allowed the identification by liquid chromatography-tandem mass spectrometry and searches in the full NCBInr (National Center for Biotechnology Information nonredundant) database of 66 proteins representing 36 different proteins validated through Peptide and Protein Prophet of Scaffold software.

  15. Primary culture of adult rat liver cells. I. Preparation of isolated cells from trypsin-perfused liver of adult rat

    Directory of Open Access Journals (Sweden)

    Miyazaki,Masahiro

    1977-12-01

    Full Text Available Isolated hepatic cells from adult rats were prepared by perfusing the livers with trypsin. The highest yield of viable cells was obtained by perfusing the liver with 0.1% trypsin, pH 7.0, at 37 degrees C for 30 min. Following this treatment about 70% of cells excluded trypan blue. The isolated cells contained many binucleate cells. Between 60 and 70% of DNA present originally in the liver was recovered from the isolated hepatic cells, which had higher glucose 6-phosphatase activity than the liver. Thus the resulting cell population seems to be rich in hepatocytes. The isolated hepatic cells, however, lost some of their cellular proteins such as alanine and tyrosine amino-transferases. It was suggested that the membranes of isolated hepatic cells might be damaged by both enzymatic digestion and mechanical destruction.

  16. Outsourcing competence

    NARCIS (Netherlands)

    Bergstra, J.; Delen, G.; van Vlijmen, B.

    2011-01-01

    The topic of this paper, competences needed for outsourcing, is organized by first providing a generic competence scheme, which is subsequently instantiated to the area of sourcing and outsourcing. Sourcing and outsourcing are positioned as different areas of activity, neither one of which is

  17. Japanese encephalitis virus induce immuno-competency in neural stem/progenitor cells.

    Directory of Open Access Journals (Sweden)

    Sulagna Das

    Full Text Available BACKGROUND: The low immunogenicity of neural stem/progenitor cells (NSPCs coupled with negligible expression of MHC antigens has popularized their use in transplantation medicine. However, in an inflammatory environment, the NSPCs express costimulatory molecules and MHC antigens, and also exhibit certain immunomodulatory functions. Since NSPCs are the cellular targets in a number of virus infections both during postnatal and adult stages, we wanted to investigate the immunological properties of these stem cells in response to viral pathogen. METHODOLOGY/PRINCIPAL FINDINGS: We utilized both in vivo mouse model and in vitro neurosphere model of Japanese encephalitis virus (JEV infection for the study. The NSPCs residing in the subventricular zone of the infected brains showed prominent expression of MHC-I and costimulatory molecules CD40, CD80, and CD86. Using Flow cytometry and fluorescence microscopy, we observed increased surface expression of co-stimulatory molecule and MHC class I antigen in NSPCs upon progressive JEV infection in vitro. Moreover, significant production of pro-inflammatory cyto/chemokines was detected in JEV infected NSPCs by Cytokine Bead Array analysis. Interestingly, NSPCs were capable of providing functional costimulation to allogenic T cells and JEV infection resulted in increased proliferation of allogenic T cells, as detected by Mixed Lymphocyte reaction and CFSE experiments. We also report IL-2 production by NSPCs upon JEV infection, which possibly provides mitogenic signals to T cells and trigger their proliferation. CONCLUSION/SIGNIFICANCE: The in vivo and in vitro findings clearly indicate the development of immunogenicity in NSPCs following progressive JEV infection, in our case, JEV infection. Following a neurotropic virus infection, NSPCs possibly behave as immunogenic cells and contribute to both the innate and adaptive immune axes. The newly discovered immunological properties of NSPCs may have implications in

  18. In vitro developmental competence of pig nuclear transferred embryos: effects of GFP transfection, refrigeration, cell cycle synchronization and shapes of donor cells.

    Science.gov (United States)

    Zhang, Yun-Hai; Pan, Deng-Ke; Sun, Xiu-Zhu; Sun, Guo-Jie; Liu, Xiao-Hui; Wang, Xiao-Bo; Tian, Xing-Hua; Li, Yan; Dai, Yun-Ping; Li, Ning

    2006-08-01

    The present study was designed to evaluate the feasibility of producing pig transgenic blastocysts expressing enhanced green fluorescent protein (GFP) and to examine the effects of shape and preparation methods of donor cells on in vitro developmental ability of pig nuclear transferred embryos (NTEs). In experiment 1, the effect of GFP transfection on development of pig NTEs was evaluated. The cleavage and blastocyst rates showed no significant difference between NTEs derived from transfected and non-transfected donors. In experiment 2, the effect of different nuclear donor preparation methods on in vitro development of NTEs was examined. The cleavage rate showed no statistically significant differences among three preparation methods. The blastocyst rates of donor cells treated once at -4 degrees C and those of freshly digested cells were similar to each other (26.3% vs 17.9%). The lowest blastocyst rates (5.88%) were observed when cells cryopreserved at -196 degrees C were used as donors. In experiment 3, the effect of different cell cycle synchronization methods on the in vitro development potential of pig NTEs was evaluated. The cleavage rate of NTEs derived from cycling cells was much better than that of NTEs derived from serum-starved cells (64.4% vs 50.5%, p refrigerated pig GFP-transfected cells could be used as donors in nuclear transfer and these NTEs could be effectively developed to blastocyst stage; (ii) serum starvation of GFP-transfected cells is not required for preimplantation development of pig NTEs; and (iii) a rough surface of GFP-transfected donor cells affects fusion rate negatively but has no influence on the cleavage rate or blastocyst rate of pig NTEs.

  19. Acquiring Metastatic Competence by Oral Squamous Cell Carcinoma Cells Is Associated with Differential Expression of α-Tubulin Isoforms

    Directory of Open Access Journals (Sweden)

    Becky Lou

    2012-01-01

    Full Text Available We performed comparative global proteomics analyses of patient-matched primary (686Tu and metastatic (686Ln OSCC cells. The metastatic OSCC 686Ln cells showed greater in vitro migratory/invasive potential and distinct cell shape from their parental primary 686Tu cells. Ettan DIGE analysis revealed 1316 proteins spots in both cell lines with >85% to be quantitatively similar (<2 folds between the two cell lines. However, two protein spots among four serial spots were highly dominant in 686Ln cells. Mass spectrometry sequencing demonstrated all four spots to be α-tubulin isotypes. Further analysis showed no significant quantitative difference in the α-tubulin between the two cell lines either at mRNA or protein levels. Thus, two distinct isoforms of α-tubulin, probably due to posttranslational modification, were associated with metastatic 686Ln cells. Immunofluorescence demonstrated remarkable differences in the cytosolic α-tubulin distribution patterns between the two cells. In 686Tu cells, α-tubulin proteins formed a normal network composed of filaments. In contrast, α-tubulin in 686Ln cells exhibited only partial cytoskeletal distribution with the majority of the protein diffusely distributed within the cytosol. Since α-tubulin is critical for cell shape and mobility, our finding suggests a role of α-tubulin isoforms in acquisition of metastatic phenotype and represents potential target for therapeutic intervention.

  20. Timespacing competence

    DEFF Research Database (Denmark)

    Laursen, Helle Pia; Mogensen, Naja Dahlstrup

    2016-01-01

    Drawing on Kramsch’s (2009) conceptualization of the multilingual subject and the symbolic self, in this paper, we explore how multilingual children re-signify three intertwined myths about the bilingual student, linguistic diversity and language competence, when, in the researcher-generated acti......Drawing on Kramsch’s (2009) conceptualization of the multilingual subject and the symbolic self, in this paper, we explore how multilingual children re-signify three intertwined myths about the bilingual student, linguistic diversity and language competence, when, in the researcher....... By perceiving competences from a subjective child perspective, we learn how children do what we call timespacing competence. On that basis, we suggest paying attention to how children themselves timespace competence by focusing (more consistently) on the subjective, social, spatial and temporal dimensions...

  1. Preparation and regulating cell adhesion of anion-exchangeable layered double hydroxide micropatterned arrays.

    Science.gov (United States)

    Yao, Feng; Hu, Hao; Xu, Sailong; Huo, Ruijie; Zhao, Zhiping; Zhang, Fazhi; Xu, Fujian

    2015-02-25

    We describe a reliable preparation of MgAl-layered double hydroxide (MgAl-LDH) micropatterned arrays on gold substrate by combining SO3(-)-terminated self-assembly monolayer and photolithography. The synthesis route is readily extended to prepare LDH arrays on the SO3(-)-terminated polymer-bonded glass substrate amenable for cell imaging. The anion-exchangeable MgAl-LDH micropattern can act both as bioadhesive region for selective cell adhesion and as nanocarrier for drug molecules to regulate cell behaviors. Quantitative analysis of cell adhesion shows that selective HepG2 cell adhesion and spreading are promoted by the micropatterned MgAl-LDH, and also suppressed by methotrexate drug released from the LDH interlayer galleries.

  2. Single cell analysis of gene expression patterns of competence development and initiation of sporulation in Bacillus subtilis grown on chemically defined media

    NARCIS (Netherlands)

    Veening, J. -W.; Smits, W. K.; Hamoen, L. W.; Kuipers, O. P.

    2006-01-01

    Aim: Understanding the basis for the heterogeneous (or bistable) expression patterns of competence development and sporulation in Bacillus subtilis. Methods and Results: Using flow cytometric analyses of various promoter-GFP fusions, we have determined the single-cell gene expression patterns of com

  3. Smooth Muscle-Like Cells Generated from Human Mesenchymal Stromal Cells Display Marker Gene Expression and Electrophysiological Competence Comparable to Bladder Smooth Muscle Cells.

    Directory of Open Access Journals (Sweden)

    Juliane Brun

    Full Text Available The use of mesenchymal stromal cells (MSCs differentiated toward a smooth muscle cell (SMC phenotype may provide an alternative for investigators interested in regenerating urinary tract organs such as the bladder where autologous smooth muscle cells cannot be used or are unavailable. In this study we measured the effects of good manufacturing practice (GMP-compliant expansion followed by myogenic differentiation of human MSCs on the expression of a range of contractile (from early to late myogenic markers in relation to the electrophysiological parameters to assess the functional role of the differentiated MSCs and found that differentiation of MSCs associated with electrophysiological competence comparable to bladder SMCs. Within 1-2 weeks of myogenic differentiation, differentiating MSCs significantly expressed alpha smooth muscle actin (αSMA; ACTA2, transgelin (TAGLN, calponin (CNN1, and smooth muscle myosin heavy chain (SM-MHC; MYH11 according to qRT-PCR and/or immunofluorescence and Western blot. Voltage-gated Na+ current levels also increased within the same time period following myogenic differentiation. In contrast to undifferentiated MSCs, differentiated MSCs and bladder SMCs exhibited elevated cytosolic Ca2+ transients in response to K+-induced depolarization and contracted in response to K+ indicating functional maturation of differentiated MSCs. Depolarization was suppressed by Cd2+, an inhibitor of voltage-gated Ca2+-channels. The expression of Na+-channels was pharmacologically identified as the Nav1.4 subtype, while the K+ and Ca2+ ion channels were identified by gene expression of KCNMA1, CACNA1C and CACNA1H which encode for the large conductance Ca2+-activated K+ channel BKCa channels, Cav1.2 L-type Ca2+ channels and Cav3.2 T-type Ca2+ channels, respectively. This protocol may be used to differentiate adult MSCs into smooth muscle-like cells with an intermediate-to-late SMC contractile phenotype exhibiting voltage-gated ion

  4. Smooth Muscle-Like Cells Generated from Human Mesenchymal Stromal Cells Display Marker Gene Expression and Electrophysiological Competence Comparable to Bladder Smooth Muscle Cells

    Science.gov (United States)

    Brun, Juliane; Lutz, Katrin A.; Neumayer, Katharina M. H.; Klein, Gerd; Seeger, Tanja; Uynuk-Ool, Tatiana; Wörgötter, Katharina; Schmid, Sandra; Kraushaar, Udo; Guenther, Elke; Rolauffs, Bernd; Aicher, Wilhelm K.; Hart, Melanie L.

    2015-01-01

    The use of mesenchymal stromal cells (MSCs) differentiated toward a smooth muscle cell (SMC) phenotype may provide an alternative for investigators interested in regenerating urinary tract organs such as the bladder where autologous smooth muscle cells cannot be used or are unavailable. In this study we measured the effects of good manufacturing practice (GMP)-compliant expansion followed by myogenic differentiation of human MSCs on the expression of a range of contractile (from early to late) myogenic markers in relation to the electrophysiological parameters to assess the functional role of the differentiated MSCs and found that differentiation of MSCs associated with electrophysiological competence comparable to bladder SMCs. Within 1–2 weeks of myogenic differentiation, differentiating MSCs significantly expressed alpha smooth muscle actin (αSMA; ACTA2), transgelin (TAGLN), calponin (CNN1), and smooth muscle myosin heavy chain (SM-MHC; MYH11) according to qRT-PCR and/or immunofluorescence and Western blot. Voltage-gated Na+ current levels also increased within the same time period following myogenic differentiation. In contrast to undifferentiated MSCs, differentiated MSCs and bladder SMCs exhibited elevated cytosolic Ca2+ transients in response to K+-induced depolarization and contracted in response to K+ indicating functional maturation of differentiated MSCs. Depolarization was suppressed by Cd2+, an inhibitor of voltage-gated Ca2+-channels. The expression of Na+-channels was pharmacologically identified as the Nav1.4 subtype, while the K+ and Ca2+ ion channels were identified by gene expression of KCNMA1, CACNA1C and CACNA1H which encode for the large conductance Ca2+-activated K+ channel BKCa channels, Cav1.2 L-type Ca2+ channels and Cav3.2 T-type Ca2+ channels, respectively. This protocol may be used to differentiate adult MSCs into smooth muscle-like cells with an intermediate-to-late SMC contractile phenotype exhibiting voltage-gated ion channel

  5. Both Freshly Prepared and Frozen-Stored Amniotic Membrane Cells Express the Complement Inhibitor CD59

    Directory of Open Access Journals (Sweden)

    Ágnes Füst

    2012-01-01

    Full Text Available Amniotic membrane proved to be very effective tool in the treatment of a number of ocular surface diseases. The amniotic membrane, however, has to be stored before its transplantation onto the ocular surface followed by mandatory serologic control in order to exclude the transmission of certain viruses. Therefore it is most important to study if cryopreservation of the membrane affects cell surface expression of the molecules. We measured cell surface expression of CD59, a membrane-bound complement inhibitor on the cells of freshly prepared and cryopreserved amniotic membrane. Cells of amniotic membrane were separated mechanically. Epithelial and mesenchymal cells were identified by the intracellular expression of nanog and the cell surface ICAM1 positivity, respectively. Multicolor flow cytometric immunophenotyping was used for determination of the CD59 expression. CellQuest-Pro software program (Becton Dickinson was used both for measurements and analysis. CD59-positive cells could be detected in all investigated samples and in all investigated cell types, although the expression level of CD59 differed. CD59 was expressed both on freshly prepared and frozen-stored samples. Higher level of CD59 was detected on ICAM1+ mesenchymal cells than on nanog+ epithelial cells. Our findings indicate that amniotic membranes maintain their complement inhibiting capacity after cryopreservation.

  6. A simple nanostructured polymer/ZnO hybrid solar cell - preparation and operation in air

    DEFF Research Database (Denmark)

    Krebs, Frederik C; Thomann, Yi; Thomann, Ralf

    2008-01-01

    without notable loss in efficiency. The devices do not require any form of encapsulation to gain stability, while a barrier for mechanical protection may be useful. The devices are based on soluble zinc oxide nanoparticles mixed with the thermocleavable conjugated polymer poly-(3-(2-methylhexan-2-yl......A detailed description is given of the preparation of a polymer solar cell and its characterization. The solar cell can be prepared entirely in the ambient atmosphere by solution processing without the use of vacuum coating steps, and it can be operated in the ambient atmosphere with good...

  7. Dock mediates Scar- and WASp-dependent actin polymerization through interaction with cell adhesion molecules in founder cells and fusion-competent myoblasts.

    Science.gov (United States)

    Kaipa, Balasankara Reddy; Shao, Huanjie; Schäfer, Gritt; Trinkewitz, Tatjana; Groth, Verena; Liu, Jianqi; Beck, Lothar; Bogdan, Sven; Abmayr, Susan M; Önel, Susanne-Filiz

    2013-01-01

    The formation of the larval body wall musculature of Drosophila depends on the asymmetric fusion of two myoblast types, founder cells (FCs) and fusion-competent myoblasts (FCMs). Recent studies have established an essential function of Arp2/3-based actin polymerization during myoblast fusion, formation of a dense actin focus at the site of fusion in FCMs, and a thin sheath of actin in FCs and/or growing muscles. The formation of these actin structures depends on recognition and adhesion of myoblasts that is mediated by cell surface receptors of the immunoglobulin superfamily. However, the connection of the cell surface receptors with Arp2/3-based actin polymerization is poorly understood. To date only the SH2-SH3 adaptor protein Crk has been suggested to link cell adhesion with Arp2/3-based actin polymerization in FCMs. Here, we propose that the SH2-SH3 adaptor protein Dock, like Crk, links cell adhesion with actin polymerization. We show that Dock is expressed in FCs and FCMs and colocalizes with the cell adhesion proteins Sns and Duf at cell-cell contact points. Biochemical data in this study indicate that different domains of Dock are involved in binding the cell adhesion molecules Duf, Rst, Sns and Hbs. We emphasize the importance of these interactions by quantifying the enhanced myoblast fusion defects in duf dock, sns dock and hbs dock double mutants. Additionally, we show that Dock interacts biochemically and genetically with Drosophila Scar, Vrp1 and WASp. Based on these data, we propose that Dock links cell adhesion in FCs and FCMs with either Scar- or Vrp1-WASp-dependent Arp2/3 activation.

  8. KIT mutation analysis in mast cell neoplasms : recommendations of the European Competence Network on Mastocytosis

    NARCIS (Netherlands)

    Arock, M.; Sotlar, K.; Akin, C.; Broesby-Olsen, S.; Hoermann, G.; Escribano, L.; Kristensen, T. K.; Kluin-Nelemans, H. C.; Hermine, O.; Dubreuil, P.; Sperr, W. R.; Hartmann, K.; Gotlib, J.; Cross, N. C. P.; Haferlach, T.; Garcia-Montero, A.; Orfao, A.; Schwaab, J.; Triggiani, M.; Horny, H-P; Metcalfe, D. D.; Reiter, A.; Valent, P.

    Although acquired mutations in KIT are commonly detected in various categories of mastocytosis, the methodologies applied to detect and quantify the mutant type and allele burden in various cells and tissues are poorly defined. We here propose a consensus on methodologies used to detect KIT

  9. Secretory leukocyte proteinase inhibitor-competent DNA deposits are potent stimulators of plasmacytoid dendritic cells

    DEFF Research Database (Denmark)

    Skrzeczynska-Moncznik, Joanna; Wlodarczyk, Agnieszka; Zabieglo, Katarzyna

    2012-01-01

    Secretory leukocyte proteinase inhibitor (SLPI) is a well-established inhibitor of serine proteases such as human neutrophil elastase (HNE) and a NF-κB regulatory agent in immune cells. In this paper, we report that SLPI plays a previously uncharacterized role in regulating activation of plasmacy...

  10. From two competing oscillators to one coupled-clock pacemaker cell system.

    Science.gov (United States)

    Yaniv, Yael; Lakatta, Edward G; Maltsev, Victor A

    2015-01-01

    At the beginning of this century, debates regarding "what are the main control mechanisms that ignite the action potential (AP) in heart pacemaker cells" dominated the electrophysiology field. The original theory which prevailed for over 50 years had advocated that the ensemble of surface membrane ion channels (i.e., "M-clock") is sufficient to ignite rhythmic APs. However, more recent experimental evidence in a variety of mammals has shown that the sarcoplasmic reticulum (SR) acts as a "Ca(2+)-clock" rhythmically discharges diastolic local Ca(2+) releases (LCRs) beneath the cell surface membrane. LCRs activate an inward current (likely that of the Na(+)/Ca(2+) exchanger) that prompts the surface membrane "M-clock" to ignite an AP. Theoretical and experimental evidence has mounted to indicate that this clock "crosstalk" operates on a beat-to-beat basis and determines both the AP firing rate and rhythm. Our review is focused on the evolution of experimental definition and numerical modeling of the coupled-clock concept, on how mechanisms intrinsic to pacemaker cell determine both the heart rate and rhythm, and on future directions to develop further the coupled-clock pacemaker cell concept.

  11. From two competing oscillators to one coupled-clock pacemaker cell system

    Directory of Open Access Journals (Sweden)

    Yael eYaniv

    2015-02-01

    Full Text Available At the beginning of this century, debates regarding what are the main control mechanisms that ignite the action potential (AP in heart pacemaker cells dominated the electrophysiology field. The original theory which prevailed for over 50 years had advocated that the ensemble of surface membrane ion channels (i.e., M clock is sufficient to ignite rhythmic APs. However, more recent experimental evidence in a variety of mammals has shown that the sarcoplasmic reticulum (SR acts as a Ca2+ clock rhythmically discharges diastolic local Ca2+ releases (LCRs beneath the cell surface membrane. LCRs activate an inward current (likely that of the Na+/Ca2+ exchanger that prompts the surface membrane M clock to ignite an AP. Theoretical and experimental evidence has mounted to indicate that this clock crosstalk operates on a beat-to-beat basis and determines both the AP firing rate and rhythm. Our review is focused on the evolution of experimental definition and numerical modeling of the coupled-clock concept, on how mechanisms intrinsic to pacemaker cell determine both the heart rate and rhythm, and on future directions to develop further the coupled-clock pacemaker cell concept.

  12. Rapid induction of orthotopic hepatocellular carcinoma in immune-competent rats by non-invasive ultrasound-guided cells implantation

    Directory of Open Access Journals (Sweden)

    Pan Huay-Ben

    2010-07-01

    Full Text Available Abstract Background The fact that prognoses remain poor in patients with advanced hepatocellular carcinoma highlights the demand for suitable animal models to facilitate the development of anti-cancer medications. This study employed a relatively non-invasive approach to establish an orthotopic hepatocellular carcinoma model in immune-competent rats. This was done by ultrasound-guided implantation of cancer cells and the model was used to evaluate the therapeutic efficacy of short-term and low-dose epirubicin chemotherapy. Methods Rat Novikoff hepatoma cells were injected percutaneously into the liver lobes of Sprague-Dawley rats under the guidance of high resolution ultrasound. The implantation rate and the correlation between dissected and ultrasound-measured tumor sizes were evaluated. A similar induction procedure was performed by means of laparotomy in a different group of rats. Pairs of tumor measurement were compared by ultrasound and computerized tomography scan. Rats with a successful establishment of the tumor were divided into the treatment (7-day low-dose epirubicin group and the control group. The tumor sizes were non-invasively monitored by the same ultrasound machine. Blood and tumor tissues from tumor-bearing rats were examined by biochemical and histological analysis respectively. Results Ultrasound-guided implantation of Novikoff hepatoma cells led to the formation of orthotopic hepatocellular carcinoma in 60.4% (55/91 of the Sprague-Dawley rats. Moreover, tumor sizes measured by ultrasound significantly correlated with those measured by calipers after sacrificing the animals (P Conclusions Ultrasound-guided implantation of Novikoff hepatoma cells is an effective means of establishing orthotopic hepatocellular carcinoma in Sprague-Dawley rats. Short-term and low-dose epirubicin chemotherapy had perturbed tumor progression by inducing apoptosis and neovascularization blockade.

  13. Temporal restriction of pancreatic branching competence during embryogenesis is mirrored in differentiating embryonic stem cells.

    Science.gov (United States)

    Lim, Sue Mei; Li, Xueling; Schiesser, Jacqueline; Holland, Andrew M; Elefanty, Andrew G; Stanley, Edouard G; Micallef, Suzanne J

    2012-07-01

    To develop methods for the generation of insulin-producing β-cells for the treatment of diabetes, we have used GFP-tagged embryonic stem cells (ESCs) to elucidate the process of pancreas development. Using the reporter Pdx1(GFP/w) ESC line, we have previously described a serum-free differentiation protocol in which Pdx1-GFP(+) cells formed GFP bright (GFP(br)) epithelial buds that resembled those present in the developing mouse pancreas. In this study we extend these findings to demonstrate that these cells can undergo a process of branching morphogenesis, similar to that seen during pancreatic development of the mid-gestation embryo. These partially disaggregated embryoid bodies containing GFP(br) buds initially form epithelial ring-like structures when cultured in Matrigel. After several days in culture, these rings undergo a process of proliferation and form a ramified network of epithelial branches. Comparative analysis of explanted dissociated pancreatic buds from E13.5 Pdx1(GFP/w) embryos and ESC-derived GFP(br) buds reveal a similar process of proliferation and branching, with both embryonic Pdx1(GFP/w) branching pancreatic epithelium and ESC-derived GFP(br) branching organoids expressing markers representing epithelial (EpCAM and E-Cadherin), ductal (Mucin1), exocrine (Amylase and Carboxypeptidase 1A), and endocrine cell types (Glucagon and Somatostatin). ESC-derived branching structures also expressed a suite of genes indicative of ongoing pancreatic differentiation, paralleling gene expression within similar structures derived from the E13.5 fetal pancreas. In summary, differentiating mouse ESCs can generate pancreatic material that has significant similarity to the fetal pancreatic anlagen, providing an in vitro platform for investigating the cellular and molecular mechanisms underpinning pancreatic development.

  14. Preparation of cDNA libraries from vascular cells.

    Science.gov (United States)

    Lieb, M E; Taubman, M B

    1999-01-01

    The vast majority of past and present efforts in the molecular cloning of expressed sequences involve isolation of clones from cDNA libraries constructed in bacteriophage lambda (1,2). As discussed in Chapter 6 , screening these cDNA libraries using labeled probes remains the most straightforward method to isolate full length cDNAs for which some partial sequence information is known. Although the availability of high quality reagents and kits over the past decade has made the process of library construction increasingly straightforward, generation of high-quality libraries is a task that still requires a fair amount of dedicated effort. Because alternative PCR-based cloning strategies have become increasingly popular alternatives to cDNA library screening, it is useful to consider the advantages and disadvantages of each strategy before embarking on a project to construct a cDNA library (Table 1). In our opinion, it is worthwhile to construct a cDNA library when the transcript of interest is not exceedingly rare (i.e., can readily be detected by Northern blot analysis of total RNA), when multiple cDNAs will need to be cloned over a period of time, and in situations where occasional mutations can not be tolerated (for example, if the cDNA is to be expressed in mammalian cells to examine function). In situations where the transcript of interest is expressed at exceedingly low levels, or when only a single cDNA needs to be cloned, a PCR-based strategy should be considered. When the tissue source is precious (such as a unique clinical specimen), successful construction of a phage library provides a resource that can be amplified and used for multiple cloning projects over many years, but runs the risk of consuming the available RNA if the library construction fails. Table 1 Comparison of Relative Advantages of cDNA Cloning from Lambda Phage Libraries by Plaque Hybridization Compared to Newer PCR- Based Strategies Lambda phage cDNA library PCR-based strategy Freedom

  15. Exosomes released by melanoma cells prepare sentinel lymph nodes for tumor metastasis.

    Science.gov (United States)

    Hood, Joshua L; San, Roman Susana; Wickline, Samuel A

    2011-06-01

    Exosomes are naturally occurring biological nanovesicles utilized by tumors to communicate signals to local and remote cells and tissues. Melanoma exosomes can incite a proangiogenic signaling program capable of remodeling tissue matrices. In this study, we show exosome-mediated conditioning of lymph nodes and define microanatomic responses that license metastasis of melanoma cells. Homing of melanoma exosomes to sentinel lymph nodes imposes synchronized molecular signals that effect melanoma cell recruitment, extracellular matrix deposition, and vascular proliferation in the lymph nodes. Our findings highlight the pathophysiologic role and mechanisms of an exosome-mediated process of microanatomic niche preparation that facilitates lymphatic metastasis by cancer cells.

  16. Competence Building

    DEFF Research Database (Denmark)

    Borrás, Susana; Edquist, Charles

    on the one hand, and the real world of innovation policy-making on the other, typically not speaking to each other. With this purpose in mind, this paper discusses the role of competences and competence-building in the innovation process from a perspective of innovation systems; it examines how governments...... and public agencies in different countries and different times have actually approached the issue of building, maintaining and using competences in their innovation systems; it examines what are the critical and most important issues at stake from the point of view of innovation policy, looking particularly...

  17. Preparation of six quinazoline schiff bases and their inhibitory effect on HHCC and Bcap-37 cells

    Institute of Scientific and Technical Information of China (English)

    陈惠; 孙晓莉; 刘志红; 张生勇; 药立波

    2003-01-01

    Objective: To prepare six quinazoline schiff bases by six steps of chemistry organic synthesis and test their inhibitory effect on hepatomacellular carcinoma cells HHCC and mammary cancer cell Bcap-37, furthmore,to compare their antitumor activities on these two kinds of cells. Methods: 2-Amino-5-nitro-benzylcarbonitrile was the initial material, and it was under the reaction of hydrolysis, ring-closing, halogenation, addition, reduction and substitution in turn to get the six quinazoline schiff bases, MTT method was adopted to compare their anticancer activities against the two cancer cells. Result and Conclusion: Six 6-imine-4-halo substituted anilinoquinozolines were prepared. The anticancer activities against both HHCC and Bcap-37were found, furthermore, they have more potency that on HHCC than on Bcap-37. In the six compounds, the schiff base Ⅵ is the most potent compound.

  18. THE COMMUNICATIVE COMPETENCE OF FUTURE TEACHERS

    Directory of Open Access Journals (Sweden)

    Pakhomova Irina Yurevna

    2013-04-01

    Full Text Available This article describes the concept of "communicative competence of future teachers," describes the essential characteristics and features of pedagogical communication. Objective: To define the notion of "communicative competence of future teachers' Methodology of work: competence approach. Scope of the results: the preparation of future teachers at the Pedagogical University. Results: This article describes the concept of "communicative competence of future teachers," describes the essential characteristics and features of pedagogical communication.

  19. From two competing oscillators to one coupled-clock pacemaker cell system

    Science.gov (United States)

    Yaniv, Yael; Lakatta, Edward G.; Maltsev, Victor A.

    2015-01-01

    At the beginning of this century, debates regarding “what are the main control mechanisms that ignite the action potential (AP) in heart pacemaker cells” dominated the electrophysiology field. The original theory which prevailed for over 50 years had advocated that the ensemble of surface membrane ion channels (i.e., “M-clock”) is sufficient to ignite rhythmic APs. However, more recent experimental evidence in a variety of mammals has shown that the sarcoplasmic reticulum (SR) acts as a “Ca2+-clock” rhythmically discharges diastolic local Ca2+ releases (LCRs) beneath the cell surface membrane. LCRs activate an inward current (likely that of the Na+/Ca2+ exchanger) that prompts the surface membrane “M-clock” to ignite an AP. Theoretical and experimental evidence has mounted to indicate that this clock “crosstalk” operates on a beat-to-beat basis and determines both the AP firing rate and rhythm. Our review is focused on the evolution of experimental definition and numerical modeling of the coupled-clock concept, on how mechanisms intrinsic to pacemaker cell determine both the heart rate and rhythm, and on future directions to develop further the coupled-clock pacemaker cell concept. PMID:25741284

  20. Detection of opsonic antibodies against Enterococcus faecalis cell wall carbohydrates in immune globulin preparations.

    Science.gov (United States)

    Hufnagel, M; Sixel, K; Hammer, F; Kropec, A; Sava, I G; Theilacker, C; Berner, R; Huebner, J

    2014-08-01

    Three different commercially available polyvalent immune globulins (IG) were investigated for the existence of antibodies against cell wall carbohydrates of four different E. faecalis serotypes (using a cell wall carbohydrate-enzyme-linked immunosorbent assay), and whether these antibodies mediated opsonic killing (using an opsonic-killing assay). All three IG preparations contained antibodies against all four serotypes (CPS-A to CPS-D). However, only one of the three IG preparations showed opsonic killing against all four serotypes. Average killing was higher against serotypes A and B (72 and 79 %, respectively) than against serotypes C and D (30 and 37 %, respectively). Such IG preparations could play a role as an adjuvant therapeutic option in life-threatening infections with E. faecalis, particularly when resistant strains are involved.

  1. Evaluation of preparation methods for MS-based analysis of intestinal epithelial cell proteomes

    DEFF Research Database (Denmark)

    Hesselager, Marianne Overgaard; Codrea, Marius Cosmin; Bendixen, Emøke

    2015-01-01

    The gut epithelium formed between an organism and the environment plays an essential role in host–microbe interactions, yet remains one of the least characterized mammalian tissues. Especially the membrane proteins, which are critical to bacterial adhesion, are understudied, because these proteins...... are low in abundance, and large amounts of sample is needed for their preparation and for undertaking MS-based analysis. The aim of this study was to evaluate three different methods for isolation and preparation of pig intestinal epithelial cells for MS-based analysis of the proteome. Samples were...... of ease and speed of sample preparation, as well as protein recovery. In comparison, more gentle methods where intestinal epithelial cells are harvested by shaking are more time consuming, result in lower protein yield, and are prone to increased technical variation due to multiple steps involved....

  2. Technical Advance: a new cell preparation strategy that greatly improves the yield of vital and functional Tregs and NKT cells.

    Science.gov (United States)

    Rissiek, Björn; Danquah, Welbeck; Haag, Friedrich; Koch-Nolte, Friedrich

    2014-03-01

    Release of NAD(+) during preparation of murine lymphocytes causes enzymatic ADP-ribosylation of cell-surface proteins on T cells, catalyzed by toxin-related ecto-ADP-ribosyltransferase, ARTC2. ADP-riboslyation activates the cytolytic P2X7 ion channel and affects, in particular, the vitality and function of Tregs and NKT cells. Here, we describe a simple method-injection of an ARTC2-blocking nanobody-to greatly improve Treg and NKT cell vitality and to preserve their function during in vitro assays and in adoptive-transfer experiments. Moreover, we present a method for the sorting of functional, primary NKT cells, based on coexpression of ARTC2 and NK1.1. Our results pave the way for the efficient ex vivo proliferation of Tregs and NKT cells and for new experimental and therapeutic uses of these important regulatory cells.

  3. De novo epidermal regeneration using human eccrine sweat gland cells: higher competence of secretory over absorptive cells.

    Science.gov (United States)

    Pontiggia, Luca; Biedermann, Thomas; Böttcher-Haberzeth, Sophie; Oliveira, Carol; Braziulis, Erik; Klar, Agnieszka S; Meuli-Simmen, Claudia; Meuli, Martin; Reichmann, Ernst

    2014-06-01

    In our previous work, we showed that human sweat gland-derived epithelial cells represent an alternative source of keratinocytes to grow a near normal autologous epidermis. The role of subtypes of sweat gland cells in epidermal regeneration and maintenance remained unclear. In this study, we compare the regenerative potential of both secretory and absorptive sweat gland cell subpopulations. We demonstrate the superiority of secretory over absorptive cells in forming a new epidermis on two levels: first, the proliferative and colony-forming efficiencies in vitro are significantly higher for secretory cells (SCs), and second, SCs show a higher frequency of successful epidermis formation as well as an increase in the thickness of the formed epidermis in the in vitro and in vivo functional analyses using a 3D dermo-epidermal skin model. However, the ability of forming functional skin substitutes is not limited to SCs, which supports the hypothesis that multiple subtypes of sweat gland epithelial cells hold regenerative properties, while the existence and exact localization of a keratinocyte stem cell population in the human eccrine sweat gland remain elusive.

  4. New activators and inhibitors in the hair cycle clock: targeting stem cells' state of competence.

    Science.gov (United States)

    Plikus, Maksim V

    2012-05-01

    The timing mechanism of the hair cycle remains poorly understood. However, it has become increasingly clear that the telogen-to-anagen transition is controlled jointly by at least the bone morphogenic protein (BMP), WNT, fibroblast growth factor (FGF), and transforming growth factor (TGF)-β signaling pathways. New research shows that Fgf18 signaling in hair follicle stem cells synergizes BMP-mediated refractivity, whereas Tgf-β2 signaling counterbalances it. Loss of Fgf18 signaling markedly accelerates anagen initiation, whereas loss of Tgf-β2 signaling significantly delays it, supporting key roles for these pathways in hair cycle timekeeping.

  5. Evaluation of cell wall preparations for proteomics: a new procedure for purifying cell walls from Arabidopsis hypocotyls

    Directory of Open Access Journals (Sweden)

    Canut Hervé

    2006-05-01

    Full Text Available Abstract Background The ultimate goal of proteomic analysis of a cell compartment should be the exhaustive identification of resident proteins; excluding proteins from other cell compartments. Reaching such a goal closely depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific difficulties: (i the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP during the isolation procedure, (ii polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins. Several reported procedures to isolate cell walls for proteomic analyses led to the isolation of a high proportion (more than 50% of predicted intracellular proteins. Since isolated cell walls should hold secreted proteins, one can imagine alternative procedures to prepare cell walls containing a lower proportion of contaminant proteins. Results The rationales of several published procedures to isolate cell walls for proteomics were analyzed, with regard to the bioinformatic-predicted subcellular localization of the identified proteins. Critical steps were revealed: (i homogenization in low ionic strength acid buffer to retain CWP, (ii purification through increasing density cushions, (iii extensive washes with a low ionic strength acid buffer to retain CWP while removing as many cytosolic proteins as possible, and (iv absence of detergents. A new procedure was developed to prepare cell walls from etiolated hypocotyls of Arabidopsis thaliana. After salt extraction, a high proportion of proteins predicted to be secreted was released (73%, belonging to the same functional classes as proteins identified using previously described protocols. Finally, removal of intracellular proteins was obtained using detergents, but their amount represented less than 3% in mass of the total protein extract, based on protein quantification. Conclusion The

  6. Preparation and Evaluation of Multi-Layer Anodes of Solid Oxide Fuel Cell

    Science.gov (United States)

    Santiago, Diana; Farmer, Serene C.; Setlock, John A.

    2012-01-01

    The development of an energy device with abundant energy generation, ultra-high specific power density, high stability and long life is critical for enabling longer missions and for reducing mission costs. Of all different types of fuel cells, the solid oxide fuel cells (SOFC) is a promising high temperature device that can generate electricity as a byproduct of a chemical reaction in a clean way and produce high quality heat that can be used for other purposes. For aerospace applications, a power-to-weight of (is) greater than 1.0 kW/kg is required. NASA has a patented fuel cell technology under development, capable of achieving the 1.0 kW/kg figure of merit. The first step toward achieving these goals is increasing anode durability. The catalyst plays an important role in the fuel cells for power generation, stability, efficiency and long life. Not only the anode composition, but its preparation and reduction are key to achieving better cell performance. In this research, multi-layer anodes were prepared varying the chemistry of each layer to optimize the performance of the cells. Microstructure analyses were done to the new anodes before and after fuel cell operation. The cells' durability and performance were evaluated in 200 hrs life tests in hydrogen at 850 C. The chemistry of the standard nickel anode was modified successfully reducing the anode degradation from 40% to 8.4% in 1000 hrs and retaining its microstructure.

  7. Silver Vanadium Phosphorous Oxide, Ag(2)VO(2)PO(4): Chimie Douce Preparation and Resulting Lithium Cell Electrochemistry.

    Science.gov (United States)

    Kim, Young Jin; Marschilok, Amy C; Takeuchi, Kenneth J; Takeuchi, Esther S

    2011-08-15

    Recently, we have shown silver vanadium phosphorous oxide (Ag(2)VO(2)PO(4), SVPO) to be a promising cathode material for lithium based batteries. Whereas the first reported preparation of SVPO employed an elevated pressure, hydrothermal approach, we report herein a novel ambient pressure synthesis method to prepare SVPO, where our chimie douce preparation is readily scalable and provides material with a smaller, more consistent particle size and higher surface area relative to SVPO prepared via the hydrothermal method. Lithium electrochemical cells utilizing SVPO cathodes made by our new process show improved power capability under constant current and pulse conditions over cells containing cathode from SVPO prepared via the hydrothermal method.

  8. Isolation of IL-12p70-competent human monocyte-derived dendritic cells

    DEFF Research Database (Denmark)

    Søndergaard, Jonas Nørskov; Pedersen, Susanne Brix

    2012-01-01

    that moDCs generated under standard conditions develop into two subsets based on CD1a-expression with the CD1a+ moDCs being the main IL-12p70 producers. This has however not been generally accepted, which we show here because the subset described as CD1a-negative does express CD1a, but at a lower level......Diverse methodologies ranging from experimental immunological studies to immunotherapy involve the application of human monocyte-derived dendritic cells (moDCs). Considerable donor-dependent variations in the moDC production of IL-12p70 affect the outcome of these methodologies. It has been shown...

  9. Simultaneous evaluation of toxicities using a mammalian cell array chip prepared by photocatalytic lithography.

    Science.gov (United States)

    Komori, Kikuo; Nada, Jun; Nishikawa, Masaki; Notsu, Hideo; Tatsuma, Tetsu; Sakai, Yasuyuki

    2009-10-27

    A prototype of a mammalian cell array chip was developed on a flat glass surface. A superhydrophilic (water contact angle=5 degrees)/highly hydrophobic (120 degrees) pattern was prepared on a fluorinated polymer-coated glass surface by means of photocatalytic lithography, and A549 (a human alveolar epithelial cell line), Hep G2 (a human hepatoma cell line) and mouse fibroblast 3T3 cells were inoculated onto the superhydrophilic regions. The cell populations were confined in the superhydrophilic regions for at least 24 h and separated from each other for at least one week. Organ-specific toxicity of aflatoxin B(1) and non-specific toxicity of adriamycin were successfully detected by using the cell array chip.

  10. Preparation and characteristics of DNA-nanoparticles targeting to hepatocarcinoma cells

    Institute of Scientific and Technical Information of China (English)

    Qin He; Ji Liu; Xun Sun; Zhi-Rong Zhang

    2004-01-01

    AIM: To prepare thymidine kinase gene (TK gene) nanopartuckes and to investigate the expression of TK gene.METHODS: Poly(D,L-lactic-co-glycolic acid) (PLGA), a biodegradable and biocompatible polymer, was used to prepare recombinant plasmid pEGFP-AFP nanoparticles by a double-emulsion evaporation technique. Characteristics of the nanoparticles were investigated in this study, including morphology, entrapment efficiency, and tissue distribution.The expression of TK gene was also investigated by MTT assay, by which the viable cells were determined after the addition of ganciclovir (GCV). The enhanced green fluorescent protein (EGFP) expression in human hepatocellular carcinoma 5MMC-7721 cells and normal parenchymal Chang liver cells were assessed by flow cytometry.RESUITS: The prepared plasmid-nanoparticles had regular spherical surface and narrow particle size span with a mean diameter of 72±12 nm. The mean entrapment efficiency was 91.25%. A total of 80.14% DNA was found to be localized in the livers after 1-h injection with 32P-DNA-PLGA nanoparticles in mouse caudal vein. The expression of DNA encapsulated in nanopartides was much higher than that in naked DNA, and human hepatocellular carcinoma SMMC7721 cells were more sensitive to GCV than human normal parenchymal Chang liver cells.CONCLUSION: The enhanced transfection efficiency and stronger ability to protect plasmid DNA from being degraded by nucleases are due to nanoparticles encapsulation.

  11. Preparation and immobilization of noble metal nanoparticles for plasmonic solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ruoli; Pitzer, Martin; Hu, DongZhi; Schaadt, Daniel M. [Institut fuer Angewandte Physik, Karlsruher Institut fuer Technologie (KIT), Karlsruhe (Germany); DFG Centrum fuer Funktionelle Nanostrukturen (CFN), KIT (Germany); Fruk, Ljiljana [DFG Centrum fuer Funktionelle Nanostrukturen (CFN), KIT (Germany)

    2011-07-01

    Thin-film solar cells are of high interest due to good electrical properties and low material consumption. Traditional thin-film cells, however, have considerable transmission losses because of the reduced absorption volume. A promising way to enhance absorption in the active layer is the light-trapping by plasmonic nanostructures. Metallic nanoparticles have in particular shown large enhancement of the photocurrent in thin-film devices. In this poster, we present preparation of Au,Ag and Pt nanoparticles by polyol method and seed mediated methods for use in plasmonic solar cells. Polyol method typically uses ethylene glycol as the solvent and reducing agent,and in seed-mediated synthesis small nanoparticle seeds are first prepared and then used to promote the growth of different shapes of nanoparticles. We particularly focus on the use of nanocubes and nanospheres for solar cell design. Following the nanoparticle preparation, a new method to immobilize particles on GaAs surfaces via covalent chemical bonds has been developed which prevents agglomerations and allows control of the surface density. Photocurrent spectra of GaAs pin solar cells with and without particles have been recorded. These measurements show the dependence of the photocurrent enhancement on particle material, shape and density.

  12. Preparation and Primary Culture of Liver Cells Isolated from Adult Rats by Dispase Perfusion

    Directory of Open Access Journals (Sweden)

    Wahid,Syarifuddin

    1984-06-01

    Full Text Available The dispase perfusion technique was used to isolate liver cells from adult rats. The optimum conditions for obtaining many isolated liver cells with high viability were an enzyme concentration of 2000 U/ml, a pH of 7.5 and a perfusion time of 20 min. The population of isolated liver cells prepared with dispase consisted of 43.6% cells with diameters less than 20 micron and 56.4% cells with diameters above 20 micron. The isolated liver cells were cultured in basal culture medium either supplemented with or without dexamethasone (1 X 10(-5M and insulin (10 micrograms/ml. The addition of hormones to the culture medium improved the attachment efficiency of the isolated liver cells and delayed the disappearance of mature hepatocytes. Epithelial-like clear cells proliferated early in primary culture even in the presence of hormones. Therefore, functioning mature hepatocytes and proliferating epithelial-like clear cells coexisted well in the hormone-containing medium. Furthermore, the number of cultured cells reached a maximal level earlier in the presence of hormones than in the absence of hormones. The level of TAT activity in primary cultured cells was higher up to 3 days after inoculation in the presence of hormones than in their absence. No difference between G6Pase activity in primary cultured cells in the presence of hormones and that in the absence of hormones was found.

  13. Preparation of corncob grits as a carrier for immobilizing yeast cells for ethanol production.

    Science.gov (United States)

    Lee, Sang-Eun; Lee, Choon Geun; Kang, Do Hyung; Lee, Hyeon-Yong; Jung, Kyung-Hwan

    2012-12-01

    In this study, DEAE-corncobs [delignified corncob grits derivatized with 2-(diethylamino)ethyl chloride hydrochloride (DEAE·HCl)] were prepared as a carrier to immobilize yeast (Saccharomyces cerevisiae) for ethanol production. The immobilized yeast cell reactor produced ethanol under optimized DEAE·HCl derivatization and adsorption conditions between yeast cells and the DEAE-corncobs. When delignified corncob grit (3.0 g) was derivatized with 0.5M DEAE·HCl, the yeast cell suspension (OD600 = 3.0) was adsorbed at >90% of the initial cell OD600. This amount of adsorbed yeast cells was estimated to be 5.36 mg-dry cells/g-DEAE corncobs. The Qmax (the maximum cell adsorption by the carrier) of the DEAE-corncobs was estimated to be 25.1 (mg/g), based on a Languir model biosorption isotherm experiment. When we conducted a batch culture with medium recycling using the immobilized yeast cells, the yeast cells on DEAE-corncobs produced ethanol gradually, according to glucose consumption, without cells detaching from the DEAE-corncobs. We observed under electron microscopy that the yeast cells grew on the surface and in the holes of the DEAEcorncobs. In a future study, DEAE-corncobs and the immobilized yeast cell reactor system will contribute to bioethanol production from biomass hydrolysates.

  14. Crosslinked gelatin nanofibres: Preparation, characterisation and in vitro studies using glial-like cells

    Energy Technology Data Exchange (ETDEWEB)

    Tonda-Turo, C. [Politecnico di Torino, Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, Torino (Italy); Cipriani, E. [Nanostructured Interfaces and Surfaces (NIS) Centre of Excellence, Department of Chemistry IFM, Università di Torino, Via P. Giuria 7, Torino (Italy); Gnavi, S. [Department Of Human and Animal Biology, Università di Torino, Via Accademia Albertina, 23, Torino (Italy); Chiono, V.; Mattu, C.; Gentile, P. [Politecnico di Torino, Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, Torino (Italy); Perroteau, I. [Department Of Human and Animal Biology, Università di Torino, Via Accademia Albertina, 23, Torino (Italy); Zanetti, M. [Nanostructured Interfaces and Surfaces (NIS) Centre of Excellence, Department of Chemistry IFM, Università di Torino, Via P. Giuria 7, Torino (Italy); Ciardelli, G., E-mail: gianluca.ciardelli@polito.it [Politecnico di Torino, Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, Torino (Italy); CNR-IPCF UOS Pisa Via Moruzzi, 1, 56124 Pisa (Italy)

    2013-07-01

    Gelatin (GL) nanofibrous matrices mimicking the complex biological structure of the natural extracellular matrix (ECM) were prepared from aqueous solutions by electrospinning technique. GL nanofibres with a diameter size of around 300 nm were obtained optimising the process and solution parameters. To increase the GL stability in aqueous environment γ-glycidoxypropyltrimethoxysilane (GPTMS) was used as GL crosslinker. GPTMS crosslinking did not modify the nanofibrous matrix morphology: fibre diameter and membrane pores size were 327 ± 45 nm and 1.64 ± 0.37 μm, respectively. The produced GPTMS crosslinked GL nanofibres (GL/GPTMS{sub N}F) were found to support the in vitro adhesion, proliferation and survival of neonatal olfactory bulb ensheating cells (NOBECs). - Highlights: • Gelatin nanofibres were prepared from aqueous solution. • A silane-coupling agent was used as gelatin crosslinker. • Glial-like cells adhered and proliferated on the developed nanofibres. • Elongated morphology of glial-like cells was observed.

  15. New methodology of preparation support for solid oxide fuel cells using different pore forming agent

    Energy Technology Data Exchange (ETDEWEB)

    Fiuza, Raigenis da P.; Guedes, Bruna C.F.; Silva, Marcos A. da; Carvalho, Luiz F.V. de; Boaventura, Jaime S. [Universidade Federal da Bahia (IQ/UFBA), Salvador, BA (Brazil). Inst. de Quimica; Pontes, Luiz A.M. [Universidade Federal da Bahia (EP/UFBA), Salvador, BA (Brazil). Escola Politecnica. Programa de Pos-Graduacao em Engenharia Quimica

    2008-07-01

    The development of environment-friendly energy sources has been of the most important scientific and technological area. Solid oxide fuel cells (SOFC) are very promising alternative for their ability to handle renewable fuels with low emissions and high efficiency. However, this device requires massive improvement before commercial application. This work studies the pore formation in the cell anode and cathode with NaHCO{sub 3} or citric acid, comparing to graphite. The three agents make pore with similar features, but the use of NaHCO{sub 3} and citric acid considerably improves the adhesion of the electrode-electrolyte interface, critical characteristic for good cell efficiency. The prepared anode-electrolyte-cathode structure was studied by SEM technique. The SOFC prepared using citric acid was tested with gaseous ethanol, natural gas and hydrogen. For all these three fuels the SOFC shows virtually no overpotential, indicating the good ionic conductance of the electrodes-electrolyte interface.. (author)

  16. Preparation of a hydrophobic polythiophene film to improve protein adsorption and proliferation of PC 12 cells.

    Science.gov (United States)

    Li, Da-Feng; Wang, Hua-Jie; Fu, Jian-Xi; Wang, Wei; Jia, Xue-Shun; Wang, Jin-Ye

    2008-12-25

    High quality films of polythiophene with different alkyl side chains were successfully synthesized by a novel method in the presence of sodium dodecylbenzenesulonate (SDBS) under N2 atmosphere on the PTFE (polytetrafluorethylene) substrate. The as-prepared films were characterized by scanning electron microscopy (SEM), conductivity measurement, and water contact angle measurement. The morphologies of the films were homogeneous with micro-/nanostructures, and their conductivities were high enough for biomedical applications. Hydrophobicity of the films could be adjusted easily by inducing alkyl side chains with different length, which could control protein adsorption in succession. Hydrophobic polythiophene film with a long alkyl side chain had a higher ability of protein adsorption and PC 12 cell proliferation. The biocompatibility study of the synthesized films in vitro proved that the synthesized films were not cytotoxic to two cell lines used and could support cell attachment and proliferation well. Polythiophenes films prepared by in-situ deposition will be good candidates for biomedical applications.

  17. A simple pressure cell and delivery system for the preparation of Xe derivatives for protein crystallography

    Energy Technology Data Exchange (ETDEWEB)

    Stowell, M.H.; Soltis, S.M.; Kisker, C.; Peters, J.W.; Schindelin, H.; Rees, D.C.; Cascio, D.; Beamer, L.; Hart, J.; Whitby, F.G. [Carl F. and Winifred H. Braun Laboratories, Mail Stop 147-75CH, California Institute of Technology, Pasadena, CA 91125 (United States)]|[Stanford Synchrotron Radiation Laboratory, SLAC, P. O. Box 4349, Bin 69, Stanford University, CA 94309 (United States)]|[University of California Los Angeles, Institute of Molecular Biology, Department of Chemistry and Biochemistry, Los Angeles, CA 90024 (United States)]|[University of Utah Medical Center, Department of Biochemistry, 50 North Medical Drive, Salt Lake City, UT 84132 (United States)

    1996-09-01

    We have developed a simple device for preparing Xe derivatives under moderate gaseous pressure (1{endash}100 atm). The device employs a Cajon ultra-torr fitting to ensure a gas-tight seal around a standard x-ray capillary. As such, the cell can accommodate standard x-ray capillaries up to 1.5 mm in diameter without any modification. The device is straightforward to utilize, and samples can be mounted and pressurized in a matter of seconds. In addition, a simple and safe purging and pressurization system has been designed and constructed for the use at beamline 7-1 at the Stanford Synchrotron Radiation Laboratory (SSRL). We describe the construction of both the pressure cell and the delivery system and present results on the cells use in the preparation of xenon derivatives. {copyright} {ital 1996 American Institute of Physics.}

  18. Antiproliferative activity of various Uncaria tomentosa preparations on HL-60 promyelocytic leukemia cells.

    Science.gov (United States)

    Pilarski, Radosław; Poczekaj-Kostrzewska, Magdalena; Ciesiołka, Danuta; Szyfter, Krzysztof; Gulewicz, Krzysztof

    2007-01-01

    The woody Amazonian vine Uncaria tomentosa (cat's claw) has been recently more and more popular all over the world as an immunomodulatory, antiinflammatory and anti-cancer remedy. This study investigates anti-proliferative potency of several cat's claw preparations with different quantitative and qualitative alkaloid contents on HL-60 acute promyelocytic human cells by applying trypan blue exclusion and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay (MTT). By standardization and statistical comparison of the obtained results pteropodine and isomitraphylline are indicated to be most suitable for standardization of medical cat's claw preparations.

  19. Preparation of yttria-stabilized zirconia films for solid oxide fuel cells by electrophoretic deposition method

    Energy Technology Data Exchange (ETDEWEB)

    Ishihara, Tatsumi; Sato, Keiji; Mizuhara, Yukako; Takita, Yusaku (Oita Univ. (Japan). Faculty of Engineering)

    1992-06-01

    The electrophoretic deposition (EPD) method was applied for the preparation of yttria-stabilized zirconia (YSZ) films for a solid oxide fuel cell (SOFC). Dense YSZ films with uniform thickness can be readily prepared by EPD method. When the planar SOFC was fabricated by using La[sub 0.6]Sr[sub 0.4]MnO[sub 3] as a cathode and electroless plating Pt as an anode, the open circuit voltage and the maximum power density attained were 1.03 V and 1.87 W cm[sup -2], respectively. (author).

  20. Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2

    Institute of Scientific and Technical Information of China (English)

    JIANG Qiang; ZHANG Su-zhan; PENG Jia-ping; WANG Xu-lin

    2005-01-01

    Objective: To prepare microencapsulated cells releasing human tissue inhibitor ofmetalloproteinase-2 (TIMP-2), and investigate their biological characteristics in vitro. Methods: Chinese hamster ovary (CHO) cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide) assay. Enzyme linked immunosorbent assay (ELISA) and reverse zymography were used to confirm the release of biologically active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl sulfoxide (DMSO) as preservative agent. Results: The microcapsules appeared like a sphere kept proliferating over the 6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse zymography confirmed the bioactivity of MMP (matrix metalloproteinase) inhibition of TIMP-2.The cryopreservation process did not damage the microcapsule morphology nor the viability of the cells inside. Conclusion:Microencapsulated engineered CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules.

  1. Octanoate in Human Albumin Preparations Is Detrimental to Mesenchymal Stromal Cell Culture

    Directory of Open Access Journals (Sweden)

    Way-Wua Wong

    2015-01-01

    Full Text Available Cell therapies hold great promise as the next major advance in medical treatment. To enable safe, effective ex vivo culture whilst maintaining cell phenotype, growth media constituents must be carefully controlled. We have used a chemically defined mesenchymal stromal cell culture medium to investigate the influence of different preparations of human serum albumin. We examined two aspects of cell culture, growth rate as measured by population doubling time and colony forming ability which is a representative measure of the stemness of the cell population. Albumin preparations showed comparative differences in both of these criteria. Analysis of the albumin bound fatty acids also showed differences depending on the manufacturing procedure used. We demonstrated that octanoate, an additive used to stabilize albumin during pasteurization, slows growth and lowers colony forming ability during ex vivo culture. Further to this we also found the level of Na+/K+ ATPase, a membrane bound cation pump inhibited by octanoate, is increased in cells exposed to this compound. We conclude that the inclusion of human serum albumin in ex vivo growth media requires careful consideration of not only the source of albumin, but also the associated molecular cargo, for optimal cell growth and behavior.

  2. Preparation and characterization of novel polymeric microcapsules for live cell encapsulation and therapy.

    Science.gov (United States)

    Chen, Hongmei; Ouyang, Wei; Jones, Mitchell; Metz, Terrence; Martoni, Christopher; Haque, Tasima; Cohen, Rebecca; Lawuyi, Bisi; Prakash, Satya

    2007-01-01

    This article describes the preparation and in vitro characterization of novel genipin cross-linked alginate-chitosan (GCAC) microcapsules that have potential for live cell therapy applications. This microcapsule system, consisting of an alginate core with a covalently cross-linked chitosan membrane, was formed via ionotropic gelation between calcium ions and alginate, followed by chitosan coating by polyelectrolyte complexation and covalent cross-linking of chitosan by naturally derived genipin. Results showed that, using this design concept and the three-step procedure, spherical GCAC microcapsules with improved membrane strength, suppressed capsular swelling, and suitable permeability can be prepared. The suitability of this novel membrane formulation for live cell encapsulation was evaluated, using bacterial Lactobacillus plantarum 80 (pCBH1) (LP80) and mammalian HepG2 as model cells. Results showed that capsular integrity and bacterial cell viability were sustained 6 mo postencapsulation, suggesting the feasibility of using this microcapsule formulation for live bacterial cell encapsulation. The metabolic activity of the encapsulated HepG2 was also investigated. Results suggested the potential capacity of this GCAC microcapsule in cell therapy and the control of cell signaling; however, further research is required.

  3. Preliminary study on preparation of E.coli cell-free system for protein expression

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    In the new era of "Omics",the traditional techniques of protein expression in vivo can not come up with the exponential increase of genetic information.The cellfree protein synthesis system provides a new strategy of protein expression with advantages of rapid,convenient and high-throughput expression.The preparation of cell extracts,the optimization of substrate concentrations and the energy regeneration system are the key factors for the successful construction of cell-free protein expression system.In this work,the cell extract was prepared from RNase I- defective strain E.coli A19.The cell growth phase,the pressure for cell disruption and the storage condition of cell extracts were optimized.Meanwhile,the optimal substrate concentrations and the energy regeneration system were selected.Under the optimized conditions,the green fluorescent protein (GFP) reporter gene was expressed in the E.coli cell-free system with high expression level (Ca.154 μg/mL) which was 29 times higher than the expression level before optimization.

  4. Imaging mammalian cells with soft x rays: The importance of specimen preparation

    Energy Technology Data Exchange (ETDEWEB)

    Brown, J.T.; Meyer-Ilse, W. [Ernest Orlando Lawrence Berkeley National Lab., CA (United States)

    1997-04-01

    Studies of mammalian cell structure and spatial organization are a very prominent part of modern cell biology. The interest in them as well as their size make them very accommodating subject specimens for imaging with soft x-rays using the XM-1 transmission microscope built and operated by The Center for X-ray Optics on Beam Line 6.1 at the Advanced Light Source. The purpose of these experiments was to determine if the fixative protocols normally used in electron or visible light microscopy were adequate to allow imaging cells, either fibroblasts or neurons, with minimal visible radiation damage due to imaging with soft x-rays at 2.4 nm. Two cell types were selected. Fibroblasts are easily cultured but fragile cells which are commonly used as models for the detailed study of cell physiology. Neurons are complex and sensitive cells which are difficult to prepare and to culture for study in isolation from their connections with surrounding cells. These cell types pose problems in their preparation for any microscopy. To improve the contrast and to prevent postmortem alteration of the chemistry and hence the structure of cells extracted from culture or from living organisms, fixation and staining techniques are employed in electron and visible light microscopy. It has been accepted by biologists for years that these treatments create artifacts and false structure. The authors have begun to develop protocols for specimens of each of these two cell types for soft x-ray microscopy which will preserve them in as near normal state as possible using minimal fixation, and make it possible to image them in either a hydrated or dried state free of secondary addition of stains or other labels.

  5. Forming key competences from the position of competence approach

    Directory of Open Access Journals (Sweden)

    Latypova E. R.

    2012-06-01

    Full Text Available Teaching foreign languages pursues practical, educational, bringing-up and developing aims in non-linguistic educational institution. The practical aim in teaching foreign languages is preparation of a highly trained specialist, which masters key competences and can work hard on his specialty according to world standards and is ready to act in professional communication with foreigners. New requirements direct the system of education toward competence approach. The main aim of competence approach in teaching foreign languages is an intensifying of practical studying.

  6. Preparations for low-cost silica substrate of CIGS solar cell

    Science.gov (United States)

    Hsu, Ming-Seng; Chang, Chung Chih; Cheng, Hsiang Hshi; Ouyang, Yueh; Der Sheu, Shinn

    2008-08-01

    The production of CuInGaSe2 (CIGS) solar cell is based on vacuum processes, which requires a high manufacturing temperature and high cost. Our result show a simple method has been developed to prepare the silica substrates of CIGS solar cell. It's synthesized by sol-gel process from tetraethylorthosilicate (TEOS), methanol (CH3OH) and pure water (both ion-exchange and distillation) in the presence of ammonia as catalyst. The preparation procedure was elaborated as the flexible sequence to control chemical composition and properties of the particles in sol-gel-derived silica substrate. The morphology, particle size, and size distribution of CIGS substrate were characterized with dynamic light scattering (DLS) and atomic force microscopy (AFM). The results of AFM morphology and statistic evidence we find an easy way, non-vacuum and low temperature processes, to successfully prepare the CIGS solar cell substrates with surface roughness below 3 nm. It is powerful the advance study in low cost solar cell.

  7. Preparation of PLLA/bpV(pic) microspheres and their effect on nerve cells.

    Science.gov (United States)

    Lin, Qiang; Chen, Hai-yun; Li, Hao-shen; Cai, Yang-ting

    2014-02-01

    In this study, we prepared PLLA/bpV(pic) microspheres, a bpV(pic) controlled release system and examined their ability to protect nerve cells and promote axonal growth. PLLA microspheres were prepared by employing the o/w single emulsification-evaporation technique. Neural stem cells and dorsal root ganglia were divided into 3 groups in terms of the treatment they received: a routine medium group (cultured in DMEM), a PLLA microsphere group (DMEM containing PLLA microspheres alone) and a PLLA/bpV(pic) group [DMEM containing PLLA/bpV(pic) microspheres]. The effects of PLLA/bpV(pic) microspheres were evaluated by the live-dead test and measurement of axonal length. Our results showed that PLLA/bpV(pic) granulation rate was (88.2±5.6)%; particle size was (16.8±3.1)%, drug loading was (4.05±0.3)%; encapsulation efficiency was (48.5±1.8)%. The release time lasted for 30 days. In PLLA/bpV(pic) microsphere group, the cell survival rate was (95.2 ±4.77)%, and the length of dorsal root ganglion (DRG) was 718±95 μm, which were all significantly greater than those in ordinary routine medium group and PLLA microsphere group. This preliminary test results showed the PLLA/bpV(pic) microspheres were successfully prepared and they could promote the survival and growth of neural cells in DRG.

  8. Two-step protocol for preparing adherent cells for high-throughput flow cytometry.

    Science.gov (United States)

    Kaur, Mandeep; Esau, Luke

    2015-09-01

    We have developed a simple, cost-effective, and labor-efficient two-step protocol for preparing adherent cells for high-throughput flow cytometry. Adherent cells were grown on microplates, detached with 2.9 mM EDTA (pH 6.14) added directly to wells containing cell culture medium, stained, and then analyzed on a flow cytometer. This protocol bypasses washing, centrifugation, and transfer between plates, reducing the cell loss that occurs in standard multistep protocols. The method has been validated using six adherent cell lines, four commercially available dyes, and two antibodies; the results have been confirmed using two different flow cytometry (FC) instruments. Our approach has been used for estimating apoptosis, mitochondrial membrane potential, reactive oxygen species, and autophagy in response to exposure to pure compounds as well as plant and bacterial extracts.

  9. Continuous Preparation of Copper/Carbon Nanotube Composite Films and Application in Solar Cells.

    Science.gov (United States)

    Luo, Xiao Gang; Le Wu, Min; Wang, Xiao Xia; Zhong, Xin Hua; Zhao, Ke; Wang, Jian Nong

    2016-02-08

    Realizing the continuous and large scale preparation of particle/carbon nanotube (CNT) composites with enhanced functionalities, and broad applications in energy conversion, harvesting, and storage systems, remains as a big challenge. Here, we report a scalable strategy to continuously prepare particle/CNT composite films in which particles are confined by CNT films. This is achieved by the continuous condensation and deposition of a cylindrical assembly of CNTs on a paper strip and the in situ incorporation of particles during the layer-by-layer deposition process. A Cu/CNT composite film is prepared as an example; such a film exhibits very high power conversion efficiency when it is used as a counter electrode in a solar cell, compared with previous materials under otherwise identical conditions. The proposed method can be extended to other CNT-based composite films with excellent functionalities for wide applications.

  10. Preparation and evaluation of advanced catalysts for phosphoric acid fuel cells

    Science.gov (United States)

    Stonehart, P.; Baris, J.; Hockmuth, J.; Pagliaro, P.

    1984-01-01

    The platinum electrocatalysts were characterized for their crystallite sizes and the degree of dispersion on the carbon supports. One application of these electrocatalysts was for anodic oxidation of hydrogen in hot phosphoric acid fuel cells, coupled with the influence of low concentrations of carbon monoxide in the fuel gas stream. In a similar way, these platinum on carbon electrocatalysts were evaluated for oxygen reduction in hot phosphoric acid. Binary noble metal alloys were prepared for anodic oxidation of hydrogen and noble metal-refractory metal mixtures were prepared for oxygen reduction. An exemplar alloy of platinum and palladium (50/50 atom %) was discovered for anodic oxidation of hydrogen in the presence of carbon monoxide, and patent disclosures were submitted. For the cathode, platinum-vanadium alloys were prepared showing improved performance over pure platinum. Preliminary experiments on electrocatalyst utilization in electrode structures showed low utilization of the noble metal when the electrocatalyst loading exceeded one weight percent on the carbon.

  11. Influence of preparative procedures on the membrane viscoelasticity of human red cell ghosts.

    Science.gov (United States)

    Nash, G B; Tran-Son-Tay, R; Meiselman, H J

    1986-02-13

    The effects of systematic variations in the preparative procedures on the membrane viscoelastic properties of resealed human red blood cell ghosts have been investigated. Ghosts, prepared by hypotonic lysis at 0 degrees C and resealing at 37 degrees C, were subjected to: measurement of the time constant for extensional recovery (tc); measurement of the membrane shear elastic modulus (mu) via three separate techniques; determination of the membrane viscosity (eta m) via a cone-plate Rheoscope. Membrane viscosity was also determined as eta m = mu X tc. Compared to intact cells, ghosts had shorter tc, regardless of their residual hemoglobin concentration (up to 21.6 g/dl). However, prolonged exposure to hypotonic media did increase their recovery time toward the intact cell value. The shear elastic modulus, as judged by micropipette aspiration of membrane tongues (mu p), was similar for all ghosts and intact cells. This result, taken with the tc data, indicates that ghosts have reduced membrane viscosity. Rheoscopic analysis also showed that eta m was reduced for ghosts, with the degree of reduction (approx. 50%) agreeing well with that estimated by the product mu p X tc. However, flow channel and pipette elongation estimates indicated that the ghost membrane elastic modulus was somewhat elevated compared to intact cells. We conclude that: ghosts have reduced membrane viscosity; ghosts have membrane rigidities close to intact cells, except possibly when the membrane is subjected to very large strains; the reduction in eta m is not directly related to the loss of hemoglobin; prolonged exposure of ghosts to low-ionic strength media increases the membrane viscosity toward its initial cellular level. These data indicate that the mechanical characteristics of ghost membranes can be varied by changing the methods of preparation and thus have potential application to further studies of the structural determinants of red cell membrane viscoelasticity.

  12. Complete Cell Killing by Applying High Hydrostatic Pressure for Acellular Vascular Graft Preparation

    Directory of Open Access Journals (Sweden)

    Atsushi Mahara

    2014-01-01

    Full Text Available Pressure treatment has been developed in tissue engineering application. Although the tissue scaffold prepared by a ultrahydrostatic pressure treatment has been reported, an excessive pressure has a potential to disrupt a structure of extracellular matrix through protein denaturation. It is important to understand the suitable low-pressure condition and mechanisms for cell killing. In this study, cellular morphology, mitochondria activity, and membrane permeability of mammalian cells with various pressure treatments were investigated with in vitro models. When the cells were treated with a pressure of 100 MPa for 10 min, cell morphology and adherence were the same as an untreated cells. Dehydrogenase activity in mitochondria was almost the same as untreated cells. On the other hand, when the cells were treated with the pressure of more than 200 MPa, the cells did not adhere, and the dehydrogenase activity was completely suppressed. However, green fluorescence was observed in the live/dead staining images, and the cells were completely stained as red after above 500 MPa. That is, membrane permeability was disturbed with the pressure treatment of above 500 MPa. These results indicated that the pressure of 200 MPa for 10 min was enough to induce cell killing through inactivation of mitochondria activity.

  13. Thermally Sprayed Large Tubular Solid Oxide Fuel Cells and Its Stack: Geometry Optimization, Preparation, and Performance

    Science.gov (United States)

    Zhang, Shan-Lin; Li, Cheng-Xin; Liu, Shuai; Li, Chang-Jiu; Yang, Guan-Jun; He, Peng-Jiang; Yun, Liang-Liang; Song, Bo; Xie, Ying-Xin

    2017-02-01

    In this study, we develop a large tubular solid oxide fuel cells design with several cells in series on a porous cermet support, which has many characteristics such as self-sealing, low Ohmic loss, high strength, and good thermal expansion coefficient matching. Here, we investigate aspects of the cell design, manufacture, performance, and application. Firstly, the cell length and number of cells in series are optimized by theoretical analysis. Then, thermal spraying is applied as a cost-effective method to prepare all the cell components. Finally, the performance of different types of cells and two types of stacks is characterized. The maximum output power of one tube, which had 20 cells in series, reaches 31 and 40.5 W at 800 and 900 °C, respectively. Moreover, the output power of a stack assembled with 56 tubes, each with ten cells in series, reaches 800 W at 830 °C. The excellent single tube and cell stack performance suggest that thermally sprayed tubular SOFCs have significant potential for commercialized application.

  14. Thermally Sprayed Large Tubular Solid Oxide Fuel Cells and Its Stack: Geometry Optimization, Preparation, and Performance

    Science.gov (United States)

    Zhang, Shan-Lin; Li, Cheng-Xin; Liu, Shuai; Li, Chang-Jiu; Yang, Guan-Jun; He, Peng-Jiang; Yun, Liang-Liang; Song, Bo; Xie, Ying-Xin

    2017-01-01

    In this study, we develop a large tubular solid oxide fuel cells design with several cells in series on a porous cermet support, which has many characteristics such as self-sealing, low Ohmic loss, high strength, and good thermal expansion coefficient matching. Here, we investigate aspects of the cell design, manufacture, performance, and application. Firstly, the cell length and number of cells in series are optimized by theoretical analysis. Then, thermal spraying is applied as a cost-effective method to prepare all the cell components. Finally, the performance of different types of cells and two types of stacks is characterized. The maximum output power of one tube, which had 20 cells in series, reaches 31 and 40.5 W at 800 and 900 °C, respectively. Moreover, the output power of a stack assembled with 56 tubes, each with ten cells in series, reaches 800 W at 830 °C. The excellent single tube and cell stack performance suggest that thermally sprayed tubular SOFCs have significant potential for commercialized application.

  15. Solvent-Assisted Preparation of High-Performance Mesoporous CH₃NH₃Pbl₃ Perovskite Solar Cells.

    Science.gov (United States)

    Li, Zhi-Hua; Liu, Jie; Ma, Jing-Yuan; Jiang, Yan; Ge, Qian-Qing; Ding, Jie; Hu, Jin-Song; Wan, Li-Jun

    2016-01-01

    Organometal trihalide perovskite based solar cells have attracted great attention worldwide since their power conversion efficiency (PCE) have risen to over 15% within only 3 years of development. Comparing with other types of perovskite solar cells, mesostructured perovskite solar cells based on CH₃NH₃Pbl₃ as light harvesting material have already demonstrated remarkable advance in performance and reproducibility. Here, we reported a mesoscopic TiO₂/CH₃NH₃Pbl₃ heterojunction solar cell with uniform perovskite thin film prepared via solvent-assisted solution processing method. The best performing device delivered photocurrent density of 20.11 mA cm⁻², open-circuit voltage of 1.02 V, and fill factor of 0.70, leading to a PCE of 14.41%. A small anomalous hysteresis in the J-V curves was observed, where the PCE at forward scan was measured to be 84% of the PCE at reverse scan. Based on a statistical analysis, the perovskite solar cells prepared by the reported method exhibited reproducible and high PCE, indicating its promising application in the fabrication of low-cost and high-efficiency perovskite solar cells.

  16. Evaluation of a filter-syringe set for preparation of packed cell aliquots for neonatal transfusion.

    Science.gov (United States)

    Chambers, L A

    1995-09-01

    A closed-system filter-syringe set designed for preparation of prefiltered aliquots of packed red cells for neonatal transfusion was evaluated. In three experiments, filter-syringe sets were sterile-connected, and aliquots were prepared at six intervals during 35-day storage of CPDA-1 packed cells. Hemoglobin (HGB), supernatant potassium (K+), and free hemoglobin (fHGB) in the aliquot and primary storage container were compared. There was no tubing weld failure, filter-occlusion, or bacterial contamination of the units. Hemoglobin remained stable over 35 days of storage, with comparable values in the primary collection container and syringe aliquot. Supernatant potassium in the units increased to an average of 89.5 mEq/L at 35 days, and levels in the aliquots closely matched those in the primary containers at each storage interval (maximum average 91.8 mEq/L). Free hemoglobin also progressively and comparably increased in both the primary containers and syringe aliquots. The gentle negative pressure and turbulence during use of the filter-syringe set apparently causes little hemolysis. The set simplifies preparation of aliquots for neonatal small volume and syringe-pump transfusion at costs comparable to alternative preparation methods. When sterile-connected, the filter-syringe set facilitates "assigned unit" inventory management for neonatal transfusion, which has been shown to be highly effective in limiting donor exposures.

  17. Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology

    Directory of Open Access Journals (Sweden)

    Jing Ruan

    2011-02-01

    Full Text Available Jing Ruan1,*, Jie Shen2,*, Zheng Wang2, Jiajia Ji1, Hua Song1, Kan Wang1, Bin Liu1, Jinhui Li2, Daxiang Cui11Department of Bio-Nano Science and Engineering, Key Laboratory for Thin Film and Microfabrication Technology of the Ministry of Education, National Key Laboratory of Micro/Nano Fabrication Technology, Research Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 2Shanghai Institute of Digestive Diseases, Shanghai Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China. *These two authors contributed equally to this workAbstract: Efficient preparation and labeling of human induced pluripotent stem (iPS cells is a great challenge in stem cell research and development. With the aim of investigating the feasibility of using nanotechnology to enhance the preparation efficiency of iPS cells and to label iPS cells for long-term tracing and imaging, in this paper, four transcription factor genes, ie, Oct4, Sox2, LIN28, and Nanog, and packaging plasmids such as PSPAX2 and PMD2.G were cotransfected into 293T cells using Generation 5.0 polyamidoamine dendrimer-modified magnetic nanoparticles (dMNPs as a delivery system. The resultant supernatant liquids were incubated with human fibroblast cells at 37°C for 21 days, then the embryonic stem (ES cell-like clones were screened, cultured, and identified. Finally, the prepared iPS cells were labeled with fluorescent magnetic nanoparticles (FMNPs. The results showed that dMNPs can efficiently deliver all vectors into 293T cells. The resultant lentiviruses’ titers were 10-fold more than those based on Lipofectamine™ 2000. Reverse transcription polymerase chain reaction analysis showed that four genes (Oct4, Sox2, LIN28, and Nanog exhibited different expressions in iPS cells. Immunostaining analysis showed that specific surface markers of ES cells such as SSEA-3, SSEA-4, Tra-1-60, and Tra

  18. Preparation and analysis of new proton conducting membranes for fuel cells

    DEFF Research Database (Denmark)

    Lund, Peter Brilner

    2007-01-01

    A range of potential new fuel cell membranes were prepared by inserting zirconium phosphate (ZrP) into divinylbenzene (DVB) crosslinked, sulfonated, polystyrene grafted poly(ethylene-alt-tetrafluoroethylene) and poly(vinyl difluoride) membranes using an ion exchange procedure. In short, the prefo......A range of potential new fuel cell membranes were prepared by inserting zirconium phosphate (ZrP) into divinylbenzene (DVB) crosslinked, sulfonated, polystyrene grafted poly(ethylene-alt-tetrafluoroethylene) and poly(vinyl difluoride) membranes using an ion exchange procedure. In short....... Additional zirconium phosphate treatment resulted in composite ETFE samples containing up to 15 wt.% ZrP and composite PVdF samples containing up to 27 wt.%. TG analyses of the ETFE-g-PSSA and PVdF-g-PSSA composite membranes indicated no significant changes of the thermal stability in comparison...

  19. Continuous Preparation of Carbon Nanotube Film and Its Applications in Fuel and Solar Cells.

    Science.gov (United States)

    Luo, Xiao Gang; Huang, Xin Xin; Wang, Xiao Xia; Zhong, Xin Hua; Meng, Xin Xin; Wang, Jian Nong

    2016-03-01

    So far, simultaneously realizing the continuous, controllable, and scalable preparation of carbon nanotube (CNT) film has remained a big challenge. Here, we report a scalable approach to continuously prepare CNT film with good control of film size and thickness. This is achieved through the layer-by-layer condensation and deposition of a cylindrical CNT assembly that is continuously produced from a floating catalyst CVD reactor on a paper strip. The promising applications of such a film are demonstrated by directly using it as an effective protecting layer for the Pt/C catalyst in proton exchange membrane fuel cells and as an efficient counter electrode material in quantum-dot-sensitized solar cells.

  20. Preparation of silver-coated glass frit and its application in silicon solar cells

    Science.gov (United States)

    Feng, Xiang; Biyuan, Li; Yingfen, Li; Jian, Zhou; Weiping, Gan

    2016-07-01

    A simple electroless plating process was employed to prepare silver-coated glass frits for solar cells. The surface of the glass frits was modified with polyvinyl-pyrrolidone (PVP) before the electroless plating process. Infrared (IR) spectroscopy, field emission scanning electron microscopy (FESEM), and x-ray diffraction (XRD) were used to characterize the PVP modified glass frits and investigate the mechanism of the modification process. It was found that the PVP molecules adsorbed on the glass frit surface and reduced the silver ions to the silver nanoparticles. Through epitaxial growth, these nanoparticles were uniformly deposited onto the surface of the glass frit. Silicon solar cells with this novel silver coating exhibited a photoelectric conversion efficiency increase of 0.33%. Compared with the electroless plating processes, this method provides a simple route to prepare silver-coated glass frits without introducing impurity ions.

  1. Preparation of silver-coated glass frit and its application in silicon solar cells

    Institute of Scientific and Technical Information of China (English)

    向锋; 李碧渊; 黎应芬; 周健; 甘卫平

    2016-01-01

    A simple electroless plating process was employed to prepare silver-coated glass frits for solar cells. The surface of the glass frits was modified with polyvinyl-pyrrolidone (PVP) before the electroless plating process. Infrared (IR) spectroscopy, field emission scanning electron microscopy (FESEM), and x-ray diffraction (XRD) were used to characterize the PVP modified glass frits and investigate the mechanism of the modification process. It was found that the PVP molecules adsorbed on the glass frit surface and reduced the silver ions to the silver nanoparticles. Through epitaxial growth, these nanoparticles were uniformly deposited onto the surface of the glass frit. Silicon solar cells with this novel silver coating exhibited a photoelectric conversion efficiency increase of 0.33%. Compared with the electroless plating processes, this method provides a simple route to prepare silver-coated glass frits without introducing impurity ions.

  2. Preparation of electrolyte membranes for micro tubular solid oxide fuel cells

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Yttria-stabilized zirconia (YSZ) micro tubular electrolyte membranes for solid oxide fuel cells (SOFCs) were prepared via the combined wet phase inversion and sintering technique. The as-derived YSZ mi- cro tubes consist of a thin dense skin layer and a thick porous layer that can serve as the electrode of fuel cells. The dense and the porous electrolyte layers have the thickness of 3-5 μm and 70-90 μm, respectively, while the inner surface porosity of the porous layer is higher than 28.1%. The two layers are perfectly integrated together to preclude the crack or flake of electrolyte film from the electrode. The presented method possesses distinct advantages such as technological simplicity, low cost and high reliability, and thus provides a new route for the preparation of micro tubular SOFCs.

  3. Subcellular SIMS imaging of isotopically labeled amino acids in cryogenically prepared cells

    Energy Technology Data Exchange (ETDEWEB)

    Chandra, Subhash

    2004-06-15

    Ion microscopy is a potentially powerful technique for localization of isotopically labeled molecules. In this study, L-arginine and phenylalanine amino acids labeled with stable isotopes {sup 13}C and {sup 15}N were localized in cultured cells with the ion microscope at 500 nm spatial resolution. Cells were exposed to the labeled amino acids and cryogenically prepared. SIMS analyses were made in fractured freeze-dried cells. A dynamic distribution was observed from labeled arginine-treated LLC-PK{sub 1} kidney cells at mass 28 ({sup 13}C{sup 15}N) in negative secondaries, revealing cell-to-cell heterogeneity and preferential accumulation of the amino acid (or its metabolite) in the nucleus and nucleolus of some cells. The smaller nucleolus inside the nucleus was clearly resolved in SIMS images and confirmed by correlative light microscopy. The distribution of labeled phenylalanine contrasted with arginine as it was rather homogeneously distributed in T98G human glioblastoma cells. Images of {sup 39}K, {sup 23}Na and {sup 40}Ca were also recorded to confirm the reliability of sample preparation and authenticity of the observed amino acid distributions. These observations indicate that SIMS techniques can provide a valuable technology for subcellular localization of nitrogen-containing molecules in proteomics since nitrogen does not have a radionuclide tracer isotope. Amino acids labeled with stable isotopes can be used as tracers for studying their transport and metabolism in distinct subcellular compartments with SIMS. Further studies of phenylalanine uptake in human glioblastoma cells may have special significance in boron neutron capture therapy (BNCT) as a boron analogue of phenylalanine, boronophenylalanine is a clinically approved compound for the treatment of brain tumors.

  4. Subcellular SIMS imaging of isotopically labeled amino acids in cryogenically prepared cells

    Science.gov (United States)

    Chandra, Subhash

    2004-06-01

    Ion microscopy is a potentially powerful technique for localization of isotopically labeled molecules. In this study, L-arginine and phenylalanine amino acids labeled with stable isotopes 13C and 15N were localized in cultured cells with the ion microscope at 500 nm spatial resolution. Cells were exposed to the labeled amino acids and cryogenically prepared. SIMS analyses were made in fractured freeze-dried cells. A dynamic distribution was observed from labeled arginine-treated LLC-PK 1 kidney cells at mass 28 ( 13C15N) in negative secondaries, revealing cell-to-cell heterogeneity and preferential accumulation of the amino acid (or its metabolite) in the nucleus and nucleolus of some cells. The smaller nucleolus inside the nucleus was clearly resolved in SIMS images and confirmed by correlative light microscopy. The distribution of labeled phenylalanine contrasted with arginine as it was rather homogeneously distributed in T98G human glioblastoma cells. Images of 39K, 23Na and 40Ca were also recorded to confirm the reliability of sample preparation and authenticity of the observed amino acid distributions. These observations indicate that SIMS techniques can provide a valuable technology for subcellular localization of nitrogen-containing molecules in proteomics since nitrogen does not have a radionuclide tracer isotope. Amino acids labeled with stable isotopes can be used as tracers for studying their transport and metabolism in distinct subcellular compartments with SIMS. Further studies of phenylalanine uptake in human glioblastoma cells may have special significance in boron neutron capture therapy (BNCT) as a boron analogue of phenylalanine, boronophenylalanine is a clinically approved compound for the treatment of brain tumors.

  5. Plasmacytoid Dendritic Cells Act as the Most Competent Cell Type in Linking Antiviral Innate and Adaptive Immune Responses

    Institute of Scientific and Technical Information of China (English)

    Zheng Zhang; Fu-Sheng Wang

    2005-01-01

    Appropriate in vivo control of plasmacytoid dendritic cell (pDC) recruitment and activation is a fundamental requirement for defense against viral infection. During this process, a pivotal event that influences the outcome of viral infection is the production of high levels of type I interferon by pDCs. In particular, recent research findings showed that pDCs not only shape the nature of innate resistance, but are also responsible for the successful transition from innate to adaptive immunity for viral resistance. In addition, pDCs can differentiate into antigen presenting cells that may regulate tolerance to a given pathogen. Importantly, in a series of recent clinical studies,pDCs appeared to be defective in number and function in conditions of chronic viral diseases such as infected with HIV-1, HBV or HCV. pDC-associated clinical antiviral therapy is also emerging. This review describes research findings exanining the functional and antiviral properties of in vivo pDC plasticity.

  6. Hydrophilic PCU scaffolds prepared by grafting PEGMA and immobilizing gelatin to enhance cell adhesion and proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Changcan; Yuan, Wenjie; Khan, Musammir; Li, Qian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Feng, Yakai, E-mail: yakaifeng@tju.edu.cn [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin) Tianjin 300072 (China); Yao, Fanglian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Zhang, Wencheng, E-mail: wenchengzhang@yahoo.com [Department of Physiology and Pathophysiology, Logistics University of Chinese People' s Armed Police Force, Tianjin 300162 (China)

    2015-05-01

    Gelatin contains many functional motifs which can modulate cell specific adhesion, so we modified polycarbonate urethane (PCU) scaffold surface by immobilization of gelatin. PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatins onto the surface of aminated PCU scaffolds. To increase the immobilization amount of gelatin, poly(ethylene glycol) methacrylate (PEGMA) was grafted onto PCU scaffolds by surface initiated atom transfer radical polymerization. Then, following amination and immobilization, PCU-g-PEGMA-g-gelatin scaffolds were obtained. Both modified scaffolds were characterized by chemical and biological methods. After immobilization of gelatin, the microfiber surface became rough, but the original morphology of scaffolds was maintained successfully. PCU-g-PEGMA-g-gelatin scaffolds were more hydrophilic than PCU-g-gelatin scaffolds. Because hydrophilic PEGMA and gelatin were grafted and immobilized onto the surface, the PCU-g-PEGMA-g-gelatin scaffolds showed low platelet adhesion, perfect anti-hemolytic activity and excellent cell growth and proliferation capacity. It could be envisioned that PCU-g-PEGMA-g-gelatin scaffolds might have potential applications in tissue engineering artificial scaffolds. - Graphical abstract: PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatin onto the surface of aminated PCU scaffolds (method a). To increase the immobilization amount of gelatin, PEGMAs were grafted onto the scaffold surface by SI-ATRP. PCU-g-PEGMA-g-gelatin scaffolds were prepared by method b. The gelatin modified scaffolds exhibited high hydrophilicity, low platelet adhesion, perfect anti-hemolytic activity, and excellent cell adhesion and proliferation capacity. They might have potential applications as tissue engineering scaffolds for artificial blood vessels. - Highlights: • Hydrophilic scaffolds were prepared by grafting PEGMA and immobilization of gelatins. • Grafting PEGMA enhanced the immobilization amount of gelatin

  7. Preparation, Characterization, and Efficacy of Cell Wall and Ribosomal Vaccines from Legionella Pneumophila

    Science.gov (United States)

    1981-05-01

    isolated from Washington strain organisms (15). Mice either passively immunized with goat antibodies to an attenuated strain or actively immunized with...strains differ in major antigenic components. Antibodies produced by L. pneumophila have been shown to confer passive immunity (15, 31) and to have...It J 1- %0. U %o .T- ON_ C-1. 777 22 TABLE 2. Protection afforded AICR/J irice (n = /gou) inmunized with cell wall And ribosome vaccines prepared from

  8. Preparation of Al-Sr Master Alloy in Aluminum Electrolysis Cell

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Al-Sr master alloy was prepared by using liquid aluminum cathode and a mixture of Na3AlF6- SrCO3 as the basic molten salt electrolyte in a laboratory electrolysis cell.The effects of electrolyte composition,electrolysis temperature,cathodic current density and the electrolytic duration on Sr content of Al-Sr alloy were studied.Through laboratory experiments,the parameters for smooth electrolytic reaction were proposed.

  9. Facile preparation of smooth perovskite films for efficient meso/planar hybrid structured perovskite solar cells.

    Science.gov (United States)

    Zhang, Meng; Yu, Hua; Yun, Jung-Ho; Lyu, Miaoqiang; Wang, Qiong; Wang, Lianzhou

    2015-06-21

    Smooth organolead halide perovskite films for meso/planar hybrid structured perovskite solar cells were prepared by a simple compressed air blow-drying method under ambient conditions. The resultant perovskite films show high surface coverage, leading to a device power conversion efficiency of over 10% with an open circuit voltage up to 1.003 V merely using pristine poly(3-hexylthiophene) (P3HT) as a hole transporter.

  10. Preparation of cell lines for single-cell analysis of transcriptional activation dynamics.

    Science.gov (United States)

    Rafalska-Metcalf, Ilona U; Janicki, Susan M

    2013-01-01

    Imaging molecularly defined regions of chromatin in single living cells during transcriptional activation has the potential to provide new insight into gene regulatory mechanisms. Here, we describe a method for isolating cell lines with multi-copy arrays of reporter transgenes, which can be used for real-time high-resolution imaging of transcriptional activation dynamics in single cells.

  11. Biomarker and competing endogenous RNA potential of tumor-specific long noncoding RNA in chromophobe renal cell carcinoma

    Science.gov (United States)

    He, Hai-Tao; Xu, Mu; Kuang, Ye; Han, Xiao-Yun; Wang, Ming-Qi; Yang, Qing

    2016-01-01

    Background Accumulating evidence suggests long noncoding RNAs (lncRNAs) play important roles in the initiation and progression of cancers. However, their functions in chromophobe renal cell carcinoma (chRCC) are not fully understood. Methods We analyzed the expression profiles of lncRNA, microRNA, and protein-coding RNA, along with the clinical information of 59 primary chRCC patients collected from The Cancer Genome Atlas database to identify lncRNA biomarkers for prognosis. We also constructed an lncRNA–microRNA–mRNA coexpression network (competitive endogenous RNAs network) by bioinformational approach. Results One hundred and forty-two lncRNAs were found to be differentially expressed between the cancer and normal tissues (fold change ≥1.5, P<0.001). Among them, 12 lncRNAs were also differentially expressed with the corresponding clinical characteristics (fold change ≥1.5, P<0.01). Besides, 7 lncRNAs (COL18A1-AS, BRE-AS1, SNHG7, TMEM51-AS1, C21orf62-AS1, LINC00336, and LINC00882) were identified to be significantly correlated with overall survival (log-rank P<0.05). A competitive endogenous RNA network in chRCC containing 16 lncRNAs, 18 miRNAs, and 168 protein-coding RNAs was constructed. Conclusion Our results identified specific lncRNAs associated with chRCC progression and prognosis, and presented competing endogenous RNA potential of lncRNAs in the tumor.

  12. Transduction and oncolytic profile of a potent replication-competent adenovirus 11p vector (RCAd11pGFP in colon carcinoma cells.

    Directory of Open Access Journals (Sweden)

    Jim Silver

    Full Text Available Replication-competent adenovirus type 5 (Ad5 vectors promise to be more efficient gene delivery vehicles than their replication-deficient counterparts, and chimeric Ad5 vectors that are capable of targeting CD46 are more effective than Ad5 vectors with native fibers. Although several strategies have been used to improve gene transduction and oncolysis, either by modifying their tropism or enhancing their replication capacity, some tumor cells are still relatively refractory to infection by chimeric Ad5. The oncolytic effects of the vectors are apparent in certain tumors but not in others. Here, we report the biological and oncolytic profiles of a replication-competent adenovirus 11p vector (RCAd11pGFP in colon carcinoma cells. CD46 was abundantly expressed in all cells studied; however, the transduction efficiency of RCAd11pGFP varied. RCAd11pGFP efficiently transduced HT-29, HCT-8, and LS174T cells, but it transduced T84 cells, derived from a colon cancer metastasis in the lung, less efficiently. Interestingly, RCAd11p replicated more rapidly in the T84 cells than in HCT-8 and LS174T cells and as rapidly as in HT-29 cells. Cell toxicity and proliferation assays indicated that RCAd11pGFP had the highest cell-killing activities in HT29 and T84 cells, the latter of which also expressed the highest levels of glycoproteins of the carcinoma embryonic antigen (CEA family. In vivo experiments showed significant growth inhibition of T84 and HT-29 tumors in xenograft mice treated with either RCAd11pGFP or Ad11pwt compared to untreated controls. Thus, RCAd11pGFP has a potent cytotoxic effect on colon carcinoma cells.

  13. Cell proliferation on modified DLC thin films prepared by plasma enhanced chemical vapor deposition.

    Science.gov (United States)

    Stoica, Adrian; Manakhov, Anton; Polčák, Josef; Ondračka, Pavel; Buršíková, Vilma; Zajíčková, Renata; Medalová, Jiřina; Zajíčková, Lenka

    2015-06-12

    Recently, diamondlike carbon (DLC) thin films have gained interest for biological applications, such as hip and dental prostheses or heart valves and coronary stents, thanks to their high strength and stability. However, the biocompatibility of the DLC is still questionable due to its low wettability and possible mechanical failure (delamination). In this work, DLC:N:O and DLC: SiOx thin films were comparatively investigated with respect to cell proliferation. Thin DLC films with an addition of N, O, and Si were prepared by plasma enhanced CVD from mixtures of methane, hydrogen, and hexamethyldisiloxane. The films were optically characterized by infrared spectroscopy and ellipsometry in UV-visible spectrum. The thickness and the optical properties were obtained from the ellipsometric measurements. Atomic composition of the films was determined by Rutherford backscattering spectroscopy combined with elastic recoil detection analysis and by x-ray photoelectron spectroscopy. The mechanical properties of the films were studied by depth sensing indentation technique. The number of cells that proliferate on the surface of the prepared DLC films and on control culture dishes were compared and correlated with the properties of as-deposited and aged films. The authors found that the level of cell proliferation on the coated dishes was high, comparable to the untreated (control) samples. The prepared DLC films were stable and no decrease of the biocompatibility was observed for the samples aged at ambient conditions.

  14. A Simple and Rapid Method for Preparing a Cell-Free Bacterial Lysate for Protein Synthesis

    Science.gov (United States)

    Kaduri, Maya; Shainsky-Roitman, Janna; Goldfeder, Mor; Ivanir, Eran; Benhar, Itai; Shoham, Yuval; Schroeder, Avi

    2016-01-01

    Cell-free protein synthesis (CFPS) systems are important laboratory tools that are used for various synthetic biology applications. Here, we present a simple and inexpensive laboratory-scale method for preparing a CFPS system from E. coli. The procedure uses basic lab equipment, a minimal set of reagents, and requires less than one hour to process the bacterial cell mass into a functional S30-T7 extract. BL21(DE3) and MRE600 E. coli strains were used to prepare the S30-T7 extract. The CFPS system was used to produce a set of fluorescent and therapeutic proteins of different molecular weights (up to 66 kDa). This system was able to produce 40–150 μg-protein/ml, with variations depending on the plasmid type, expressed protein and E. coli strain. Interestingly, the BL21-based CFPS exhibited stability and increased activity at 40 and 45°C. To the best of our knowledge, this is the most rapid and affordable lab-scale protocol for preparing a cell-free protein synthesis system, with high thermal stability and efficacy in producing therapeutic proteins. PMID:27768741

  15. High-throughput preparation methods of crude extract for robust cell-free protein synthesis.

    Science.gov (United States)

    Kwon, Yong-Chan; Jewett, Michael C

    2015-03-02

    Crude extract based cell-free protein synthesis (CFPS) has emerged as a powerful technology platform for high-throughput protein production and genetic part characterization. Unfortunately, robust preparation of highly active extracts generally requires specialized and costly equipment and can be labor and time intensive. Moreover, cell lysis procedures can be hard to standardize, leading to different extract performance across laboratories. These challenges limit new entrants to the field and new applications, such as comprehensive genome engineering programs to improve extract performance. To address these challenges, we developed a generalizable and easily accessible high-throughput crude extract preparation method for CFPS based on sonication. To validate our approach, we investigated two Escherichia coli strains: BL21 Star™ (DE3) and a K12 MG1655 variant, achieving similar productivity (defined as CFPS yield in g/L) by varying only a few parameters. In addition, we observed identical productivity of cell extracts generated from culture volumes spanning three orders of magnitude (10 mL culture tubes to 10 L fermentation). We anticipate that our rapid and robust extract preparation method will speed-up screening of genomically engineered strains for CFPS applications, make possible highly active extracts from non-model organisms, and promote a more general use of CFPS in synthetic biology and biotechnology.

  16. Imaging of Bacterial and Fungal Cells Using Fluorescent Carbon Dots Prepared from Carica papaya Juice.

    Science.gov (United States)

    Kasibabu, Betha Saineelima B; D'souza, Stephanie L; Jha, Sanjay; Kailasa, Suresh Kumar

    2015-07-01

    In this paper, we have described a simple hydrothermal method for preparation of fluorescent carbon dots (C-dots) using Carica papaya juice as a precursor. The synthesized C-dots show emission peak at 461 nm with a quantum yield of 7.0 %. The biocompatible nature of C-dots was confirmed by a cytotoxicity assay on E. coli. The C-dots were used as fluorescent probes for imaging of bacterial (Bacillus subtilis) and fungal (Aspergillus aculeatus) cells and emitted green and red colors under different excitation wavelengths, which indicates that the C-dots can be used as a promising material for cell imaging.

  17. Preparation and properties of microencapsulated genetically engineered bacteria cells for oral therapy of uremia

    Institute of Scientific and Technical Information of China (English)

    GAO Hong; YU Yaoting; CAI Baoli; WANG Manyan

    2004-01-01

    Microencapsulated genetically engineered bacteria cells are a novel approach of oral therapy for uremia.Klebsiella aerogenes urease genes (UreaDABCEFG) are transformed into E. coli DH5α cells through plasmid pKAU17. The transformant can use urea or ammonia as its sole nitrogen source through strain training. The urease genetically engineered bacteria cells are entrapped in polyvinyl alcohol (PVA) microcapsules, which can be used to remove urea from uremia patients. The mechanical strength of PVA microcapsules is significantly higher than that of APA microcapsules. This suggests that the problem of friability of APA can be solved in this way. The optimal conditions for the preparation of PVA microencapsulated genetically engineered bacterial cells are: polyvinyl alcohol (PVA, 2450±50)used as the carrier at a concentration 6%, the pH value of boric acid as crosslinking reagent 6.5, crosslinking time 24 h,entrapment ratio of bacteria 8%, air flow rate of the encapsulate device 3 L/min and liquid flow rate at 1 mL/10 min.The average diameter of microcapsules prepared under these optimal conditions is 20-40 mesh. Experiments in vitro showed that one hundred milligrams of wet bacterial cells in PVA microcapsules could remove 18.4 mg of urea in 4 h.

  18. Expression of HSV-1 ICP0 Antigen Peptide in Prokaryotic Cells and Preparation of Specific Antibody

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    As an immediate-early protein of herpes simplex virus, infected-cell polypeptide 0 (ICP0) exhibits complicated interactions with host cells, and its regulatory function on gene expression is of great importance. Since the ICP0 encoding sequence contains many rare codons which are absent in E.coli, and ICP0 is highly unstable in prokaryotic cells, expression of entire ICP0 in prokaryotic cells has never been reported. In order to further investigate the function of ICP0, a recombinant plasmid was constructed by subcloning a cDNA fragment encoding an amino-terminal of 105 residues of the ICP0 protein into pGEX-5x-1 vector. The resulting GST-105 fusion antigen peptide was expressed with high efficiency in E.coli. Antibodies prepared after the immunization of mice with purified fusion protein can recognize not only the denatured ICP0 protein, but also the native ICP0 protein with normal biological conformation.

  19. Sample preparation and in situ hybridization techniques for automated molecular cytogenetic analysis of white blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Rijke, F.M. van de; Vrolijk, H.; Sloos, W. [Leiden Univ. (Netherlands)] [and others

    1996-06-01

    With the advent in situ hybridization techniques for the analysis of chromosome copy number or structure in interphase cells, the diagnostic and prognostic potential of cytogenetics has been augmented considerably. In theory, the strategies for detection of cytogenetically aberrant cells by in situ hybridization are simple and straightforward. In practice, however, they are fallible, because false classification of hybridization spot number or patterns occurs. When a decision has to be made on molecular cytogenetic normalcy or abnormalcy of a cell sample, the problem of false classification becomes particularly prominent if the fraction of aberrant cells is relatively small. In such mosaic situations, often > 200 cells have to be evaluated to reach a statistical sound figure. The manual enumeration of in situ hybridization spots in many cells in many patient samples is tedious. Assistance in the evaluation process by automation of microscope functions and image analysis techniques is, therefore, strongly indicated. Next to research and development of microscope hardware, camera technology, and image analysis, the optimization of the specimen for the (semi)automated microscopic analysis is essential, since factors such as cell density, thickness, and overlap have dramatic influences on the speed and complexity of the analysis process. Here we describe experiments that have led to a protocol for blood cell specimen that results in microscope preparations that are well suited for automated molecular cytogenetic analysis. 13 refs., 4 figs., 1 tab.

  20. Are there priority competencies for preparation of accountants?: A study about Brazilian accountants ¿Existen competencias que deben ser prioridad en el desarrollo del contador?: Un estudio sobre los contadores Existem competências a serem priorizadas no desenvolvimento do contador?: Um estudo sobre os contadores brasileiros

    Directory of Open Access Journals (Sweden)

    Ricardo Lopes Cardoso

    2010-09-01

    Full Text Available This study sought to determine the need for priority of competencies in preparation of accountants as well as variations between men and women. This question is motivated by Hardern (1995, Morgan (1997, Moramed and Lashine (2003and IFAC (2003 with a theoretical foundation of competencies related to McClelland (1973, 1998 and Boyatzis (1982. Professional references are Hardern (1995, AICPA (1999 and IFAC (2003. It was based on 18 competencies that were collected from 24 articles for questions submitted to 159 accountants and the data collection instrument showed a 0.8774 Cronbach alpha. Replies were submitted to the Kruskal-Wallis test which showed a significance level of El objetivo principal de este estudio es verificar si existen competencias que deben ser prioridad en el desarrollo del contador, como también analizar si esa prioridad se altera entre hombres y mujeres. Ese cuestionamiento tiene como motivación las observaciones de Hardern (1995, Morgan (1997 y IFAC (2003. El fundamento teórico sobre competencias está relacionado al estudio de McClelland (1973,1998, Boyatzis (1982 y Spencer y Spencer (1993. Los estudios relacionados a las competencias del contador no están basados en fundamentos de la Psicología y de Recursos Humanos, y ni siquiera en el análisis de estructuras genéricas de competencias o en la determinación de prioridades en el desarrollo de competencias de los contadores, esta última es el objetivo de este trabajo. La investigación está basada en el estudio de 18 competencias del contador y fue submetida a 159 contadores. El instrumento de recolección de los datos presentó un Alfa de Cronbach de 0,8774. A partir de las respuestas se realizó el test de Kruskal-Wallis, que demostró un nivel de significancia O objetivo principal deste estudo é verificar se existem competências a serem priorizadas no desenvolvimento do contador, bem como analisar se essa prioridade se altera entre homens e mulheres. Esse

  1. Supplementation with cumulus cell masses improves the in vitro meiotic competence of porcine cumulus-oocytes complexes derived from small follicles.

    Science.gov (United States)

    Matsunaga, R; Funahashi, H

    2017-03-30

    The present study was conducted to examine the supplemented effect of cumulus cell masses (CCMs) derived from middle follicle (MF; 3-6 mm diameter) on the morphology and the meiotic or developmental competence of oocytes from small follicles (SF; 1-2 mm diameter). The number of cumulus cells surrounding oocytes just after collection was also lower in cumulus-oocyte complexes (COCs) from SF than MF. The ooplasmic diameter of oocytes was significantly smaller in SF-derived oocytes than MF-derived ones before and after in vitro maturation (IVM), whereas the diameter significantly increased during the culture. Co-culture of SF-derived COCs with MF-derived CCMs during IVM significantly improved the meiotic competence of the oocytes to the metaphase-II stage. Furthermore, the ooplasmic diameter of SF-derived COCs during IVM was increased to the similar size of MF-derived those in the presence of MF-derived CCMs. The abilities of oocytes to be penetrated, to form male pronuclear formation and to cleave or develop to the blastocyst stage were not affected by the co-culture with CCMs. Electrophoretic analysis of CCM secretions clearly showed the presence of more protein(s) approximately 27.6 kDa in the conditioned medium when supplemented with MF-derived CCMs. In conclusion, we demonstrate that supplementation with MF-derived CCMs improves the ooplasmic diameter and meiotic competence of SF-derived oocytes.

  2. Al2 O3 Underlayer Prepared by Atomic Layer Deposition for Efficient Perovskite Solar Cells.

    Science.gov (United States)

    Zhang, Jinbao; Hultqvist, Adam; Zhang, Tian; Jiang, Liangcong; Ruan, Changqing; Yang, Li; Cheng, Yibing; Edoff, Marika; Johansson, Erik M J

    2017-08-31

    Perovskite solar cells, as an emergent technology for solar energy conversion, have attracted much attention in the solar cell community by demonstrating impressive enhancement in power conversion efficiencies. However, the high temperature and manually processed TiO2 underlayer prepared by spray pyrolysis significantly limit the large-scale application and device reproducibility of perovskite solar cells. In this study, lowtemperature atomic layer deposition (ALD) is used to prepare a compact Al2 O3 underlayer for perovskite solar cells. The thickness of the Al2 O3 layer can be controlled well by adjusting the deposition cycles during the ALD process. An optimal Al2 O3 layer effectively blocks electron recombination at the perovskite/fluorine-doped tin oxide interface and sufficiently transports electrons through tunneling. Perovskite solar cells fabricated with an Al2 O3 layer demonstrated a highest efficiency of 16.2 % for the sample with 50 ALD cycles (ca. 5 nm), which is a significant improvement over underlayer-free PSCs, which have a maximum efficiency of 11.0 %. Detailed characterization confirms that the thickness of the Al2 O3 underlayer significantly influences the charge transfer resistance and electron recombination processes in the devices. Furthermore, this work shows the feasibility of using a high band-gap semiconductor such as Al2 O3 as the underlayer in perovskite solar cells and opens up pathways to use ALD Al2 O3 underlayers for flexible solar cells. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Preparation of hydrogel hollow particles for cell encapsulation by a method of polyester core degradation.

    Science.gov (United States)

    Rabanel, J-M; Hildgen, P

    2004-06-01

    Implantation of encapsulated cells in particles of less than 1 mm (micro-encapsulation) has been proposed as a cell synthesized bio-molecule delivery system. Encapsulation provides immuno-isolation, protecting foreign cells from host immune system while nutrients, oxygen and therapeutic products can diffuse freely across capsule walls. A new method is described for the synthesis of a new family of hollow microparticles for cell encapsulation. Unlike other micro-encapsulation methods, encapsulation in those devices will take place after capsule synthesis, by micro-injection. The microcapsules were prepared by a three-steps original procedure: first, synthesis of a core particle, followed by coating with a layer of epichlorohydrin cross-linked amylo-pectin gel and, finally, selective degradation of the core particle to create the cavity. Initial experiments make use of amylo-pectin cross-linked with trimetaphosphate as core particle material. However, selective degradation was difficult to achieve. In further essays, polyesters were used successfully for the preparation of core particles. Optimizations were carried out and the permeability and morphology of the hollow particles were investigated. The preliminary results show that the new method has the potential to become a standard procedure to obtain hydrogel hollow particles. Moreover, the permeability study seems to be in accordance with specifications for immuno-isolation.

  4. Efficient and stable perovskite solar cells prepared in ambient air irrespective of the humidity.

    Science.gov (United States)

    Tai, Qidong; You, Peng; Sang, Hongqian; Liu, Zhike; Hu, Chenglong; Chan, Helen L W; Yan, Feng

    2016-04-01

    Poor stability of organic-inorganic halide perovskite materials in humid condition has hindered the success of perovskite solar cells in real applications since controlled atmosphere is required for device fabrication and operation, and there is a lack of effective solutions to this problem until now. Here we report the use of lead (II) thiocyanate (Pb(SCN)2) precursor in preparing perovskite solar cells in ambient air. High-quality CH3NH3PbI3-x(SCN)x perovskite films can be readily prepared even when the relative humidity exceeds 70%. Under optimized processing conditions, we obtain devices with an average power conversion efficiency of 13.49% and the maximum efficiency over 15%. In comparison with typical CH3NH3PbI3-based devices, these solar cells without encapsulation show greatly improved stability in humid air, which is attributed to the incorporation of thiocyanate ions in the crystal lattice. The findings pave a way for realizing efficient and stable perovskite solar cells in ambient atmosphere.

  5. Efficient and stable perovskite solar cells prepared in ambient air irrespective of the humidity

    Science.gov (United States)

    Tai, Qidong; You, Peng; Sang, Hongqian; Liu, Zhike; Hu, Chenglong; Chan, Helen L. W.; Yan, Feng

    2016-01-01

    Poor stability of organic–inorganic halide perovskite materials in humid condition has hindered the success of perovskite solar cells in real applications since controlled atmosphere is required for device fabrication and operation, and there is a lack of effective solutions to this problem until now. Here we report the use of lead (II) thiocyanate (Pb(SCN)2) precursor in preparing perovskite solar cells in ambient air. High-quality CH3NH3PbI3−x(SCN)x perovskite films can be readily prepared even when the relative humidity exceeds 70%. Under optimized processing conditions, we obtain devices with an average power conversion efficiency of 13.49% and the maximum efficiency over 15%. In comparison with typical CH3NH3PbI3-based devices, these solar cells without encapsulation show greatly improved stability in humid air, which is attributed to the incorporation of thiocyanate ions in the crystal lattice. The findings pave a way for realizing efficient and stable perovskite solar cells in ambient atmosphere. PMID:27033249

  6. [Cell biology researches aboard the robotic space vehicles: preparation and performance].

    Science.gov (United States)

    Tairbekov, M G

    2006-01-01

    The article reviews the unique aspects of preparation and performance of cell biology experiments flown on robotic space vehicles Bion and Foton, and gives an overview of key findings in researches made under the author's leadership over the past decades. Described are the criteria of selecting test objects, and the conditions required for preparation and implementation of space and control (synchronous) experiments. The present-day status and issues of researches into cell responsivity to space microgravity and other factors are discussed. Also, potentialities of equipment designed to conduct experiments with cell cultures in vitro and populations of single-celled organisms are presented, as well as some ideas for new devices and systems. Unveiled are some circumstances inherent to the development and performance of space experiments, setting up laboratory facilities at the launch and landing site, and methods of safe transportation and storage of biosamples. In conclusion, the author puts forward his view on biospecies, equipment and areas of research aboard future space vehicles.

  7. Three key variables involved in feeder preparation for the maintenance of human embryonic stem cells.

    Science.gov (United States)

    Zhou, Di; Liu, Tiancheng; Zhou, Xiaoying; Lu, Guangxiu

    2009-07-01

    Although the development of a feeder-free culture system for future applications of human embryonic stem cells (hESCs), at present the regular culture system uses mitotically inactivated mouse embryonic fibroblasts (mEFs) as feeder cells for maintaining undifferentiated hESCs. Mitomycin C (MMC) is used to inactivate mEFs, but this causes DNA damage, and it is unclear whether MMC remains in the culture system after several washes. Three variables have been evaluated with respect to feeder preparation and MMC involvement, including mEF exposure to MMC, density of feeder cells, and different wash steps during the preparation of feeder cells. These variables are critical to the subsequent planting of hESCs because remnants of MMC would be unsafe with respect to long-term culture of hESCs The novel data here evaluates the remnant amounts of MMC in a hESCs culture system using HPLC/MS/MS. The ultimate objective of this study is the control of MMC within a safe range.

  8. Preparation and use of lipid microemulsions as nutritional supplements for culturing mammalian cells.

    Science.gov (United States)

    Darfler, F J

    1990-08-01

    Cells grown in vitro generally have a requirement for an exogenous source of lipid. This requirement is often met by the addition of serum, lipoproteins, or lipids complexed to albumin. To overcome the disadvantages of using lipoproteins or albumin for culturing cells in serum-free media, a method has been devised to provide necessary lipids. This report describes the preparation and use of protein-free lipid microemulsions suitable for use in tissue culture. The microemulsions are prepared from purified, synthetic lipids to produce a homogeneous, water-soluble, stable suspension that can be sterile-filtered. The best results were obtained using a sonicate of cholesterol oleate, dipalmitoyl phosphatidylcholine, dilinoleoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, cholesterol, sphingomyelin, alpha-tocopherol, alpha-tocopherol acetate, and Tween 80. Using Chinese hamster ovary (CHO) cells in a protein-free medium, cell growth was 222% vs. control (no microemulsion) in a 5-d assay. Inclusion of the microemulsion to protein-free media also increased the growth rate of murine hybridomas, H9 transformed T lymphoblasts, and human skin keratinocytes.

  9. WHO NEEDS INTERCULTURAL COMPETENCES?

    Directory of Open Access Journals (Sweden)

    Carmen – Laura ZARZU

    2013-06-01

    Full Text Available The current essay focuses on the need for formal education in the area of intercultural communication and training of intercultural competences. It builds on cultural identity and diversity literature, on the experiment conducted in the Low Countries in introducing a new topic for students from social sciences referring to intercultural communication and on reports and papers of international companies, organizations and agencies. The argument of globalization which should give equal opportunities to each and every world’s citizen adds pressure on managers dealing with multicultural teams. Intercultural competences gain importance in recruiting, while turning cultural diversity in team performance requires skills, knowledge and experience. Managing cultural diversity presupposes that people are aware, recognize, understand and deal with differences. Thus intercultural communication should be studied as a stand-alone topic or imbedded in other subjects in different forms of education or training, so people are prepared for intercultural, social and professional relationships.

  10. Preparation and characterization of sensing layers for pH detection in living plant cells

    Science.gov (United States)

    Mrazek, J.; Kasik, I.; Nekola, J.; Martan, T.; Podrazky, O.; Pospisilova, M.; Matejec, V.

    2011-05-01

    Local chemical sensing in living cells by fluorescence methods with submicron spatial resolution is in the scope of biologist because of bringing new information about biochemical processes in living matter [1]. One of the most important monitored variables is pH. Despite of progress of novel submicron probes suitable for in-situ measurement in living cells [1] and biological micro samples [2] still there is a lack off suitable opto-chemical transducers sensitive around pH 5-7 limiting development of novel fluorescence sensors. Moreover, the interaction of the immobilized transducer with the matrix can strongly affect its fluorescence properties. In our contributition the 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) fluorescence pH transducer was incorporated into organosols based on tetraethylorthosilicate (TEOS), 3-glycidoxypropyltrimethoxysilane (GPTMS) and 3-aminopropyltriethoxysilane (APTES). Formed organosols were spin-coated onto Pyrex glass substrates and thermally treated at 140°C for 4 hours. Prepared thin layers were exposed to Britton-Robinson buffers with different pH ranging from 4 to 8 pH units. Optical properties of immobilized BCECF were investigated by the mean of absorption and fluorescence spectroscopy Acquired results were compared with the properties of BCECF solutions. It was found that all matrices reduce the sensitivity of the BCECF transducer comparing to the free solution. GPTMS and APTES contained layers exhibited better mechanical properties and increase the solubility of BCECF inside prepared layers comparing to layers prepared from pure TEOS.

  11. Commonly used bowel preparations have significant and different effects upon cell proliferation in the colon: a pilot study

    Directory of Open Access Journals (Sweden)

    Riley Stuart A

    2008-11-01

    Full Text Available Abstract Background Markers of crypt cell proliferation are frequently employed in studies of the impact of genetic and exogenous factors on human colonic physiology. Human studies often rely on the assessment of tissue acquired at endoscopy. Modulation of cell proliferation by bowel preparation with oral laxatives may confound the findings of such studies, but there is little data on the impact of commonly used bowel preparations on markers of cell proliferation. Methods Crypt length, crypt cellularity and crypt cell proliferation were assessed in biopsies acquired after preparation with either Klean-Prep or Picolax. Crypt cell proliferation was assessed by whole-mount mitotic figure count, and by two different immunohistochemical (IHC labelling methods (Ki-67 and pHH3. Subsequent biopsies were obtained from the same patients without bowel preparation and similarly assessed. Parameters were compared between groups using analysis of variance and paired t-tests. Results There were significant differences in labelling indices (LI between biopsies taken after Klean-prep and those taken after Picolax preparation, for both Ki67 (p = 0.019 and pHH3 (p = 0.017. A similar trend was seen for whole-mount mitotic figure counts. Suppression or elevation of proliferation parameters by bowel preparation may mask any effect due to an intervention or disease. Conclusion Commonly used bowel preparations may have significant and different effects on crypt cell proliferation. This should be taken into account when designing studies and when considering the findings of existing studies.

  12. Preparation of conjugated polymer-based composite thin film for application in solar cell

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Yang-Yen, E-mail: yyyu@mail.mcut.edu.tw [Department of Materials Engineering, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Battery Research Center of Green Energy, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Center for Thin Film Technologies and Applications, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Chien, Wen-Chen [Department of Chemical Engineering, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Battery Research Center of Green Energy, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Ko, Yu-Hsin [Department of Materials Engineering, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Chen, Chih-Ping [Department of Materials Engineering, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Battery Research Center of Green Energy, Ming Chi University of Technology, 84 Gunjuan Road, Taishan, New Taipei City 243, Taiwan (China); Chang, Chao-Ching [Department of Chemical and Materials Engineering, Tamkang University, 151, Yingzhuan Rd., Tamsui Dist., New Taipei City 25137, Taiwan (China)

    2015-06-01

    This paper reports on the enhanced cell efficiency of structures and properties of regioregular poly(3-hexylthiophene) (P3HT)/multiwalled carbon nanotube (MWNT) hybrid materials. The prepared hybrid materials were characterized using ultraviolet–visible absorption spectroscopy, photoluminescence spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. Different concentrations of these MWNTs were suspended in polymer solutions and spin-cast onto indium tin oxide (ITO) glass. Solar cells with a device structure of ITO/poly(3,4-ethylenedioxythiophene):poly(4-styrenesulfonate) /P3HT:MWNTs/aluminum were then produced using evaporated aluminum as the back contact. The results showed that the ratio of P3HT to MWNTs considerably influenced the performance of the fabricated solar cells. The efficiency of the solar cells increased with the ratio of carbon nanotubes. Monochromatic incident photon-to-electron conversion efficiency analysis was performed and the results indicated that at the optimal P3HT/MWNTs ratio (= 1/1), the solar cells demonstrated a high-quality conversion of 2.16% with a fill factor of 42.22%, an open circuit voltage of 0.56 V, and a short circuit current of 9.12 mA/cm{sup 2}. - Highlights: • Solar cells ITO/PEDOT:PSS(DMSO)/P3HT:MWNT/Al were fabricated. • Optimal ratio of P3HT to MWNT was investigated. • Solar cell with 2.16% efficiency was obtained.

  13. Colocalization and interaction between elongasome and divisome during a preparative cell division phase in Escherichia coli.

    Science.gov (United States)

    van der Ploeg, René; Verheul, Jolanda; Vischer, Norbert O E; Alexeeva, Svetlana; Hoogendoorn, Eelco; Postma, Marten; Banzhaf, Manuel; Vollmer, Waldemar; den Blaauwen, Tanneke

    2013-03-01

    The rod-shaped bacterium Escherichia coli grows by insertion of peptidoglycan into the lateral wall during cell elongation and synthesis of new poles during cell division. The monofunctional transpeptidases PBP2 and PBP3 are part of specialized protein complexes called elongasome and divisome, respectively, which catalyse peptidoglycan extension and maturation. Endogenous immunolabelled PBP2 localized in the cylindrical part of the cell as well as transiently at midcell. Using the novel image analysis tool Coli-Inspector to analyse protein localization as function of the bacterial cell age, we compared PBP2 localization with that of other E. coli cell elongation and division proteins including PBP3. Interestingly, the midcell localization of the two transpeptidases overlaps in time during the early period of divisome maturation. Försters Resonance Energy Transfer (FRET) experiments revealed an interaction between PBP2 and PBP3 when both are present at midcell. A decrease in the midcell diameter is visible after 40% of the division cycle indicating that the onset of new cell pole synthesis starts much earlier than previously identified by visual inspection. The data support a new model of the division cycle in which the elongasome and divisome interact to prepare for cell division.

  14. Process competencies in a problem and project based learning environment

    DEFF Research Database (Denmark)

    Du, Xiangyun; Kolmos, Anette

    2006-01-01

    competencies. Consequently, engineering education is facing challenges regarding how to facilitate students with scientific-technological competencies as well as process competencies. Problem based learning (PBL) as an educational model is regarded as an effective example regarding preparing students...... with the expected professional competencies. Based on the educational practice of PBL Aalborg Model, which is characterized by problem-orientation, project-organization and team work, this paper examines the process of developing process competencies through studying engineering in a PBL environment from...

  15. Using Herbs and Spices/Preparing Sauces and Gravies. Learning Activity Pack and Instructor's Guide 5.11. Commercial Foods and Culinary Arts Competency-Based Series. Section 5: Basic Food Preparation.

    Science.gov (United States)

    Florida State Univ., Tallahassee. Center for Studies in Vocational Education.

    This document consists of a learning activity packet (LAP) for the student and an instructor's guide for the teacher. The LAP is intended to acquaint occupational home economics students with herbs and spices and the selection and preparation of sauces and gravies. Illustrated information sheets and learning activities are provided in these areas:…

  16. Biological tissue and cell culture specimen preparation for TEM nanoparticle characterization.

    Science.gov (United States)

    Nagashima, Kunio; Zheng, Jiwen; Parmiter, David; Patri, Anil K

    2011-01-01

    This chapter outlines the procedures for ex vivo TEM preparation of nanoparticle-containing tissue or cell culture samples using an epoxy resin embedding method. The purpose of this procedure is to preserve the structure of tissue in a hardened epoxy block with minimal disruption of cellular structures, to aid in the meaningful analysis of in vivo or cell culture experiments. The process begins with hydrated tissue and ends with tissue that is virtually water-free and preserved in a static state within a plastic resin matrix. The resin mixture permeates the dehydrated tissue, making the sample firm enough to cut. Procedures are also given for fixing nanoparticle-containing cell culture samples.

  17. Pt-Ru Catalysts Prepared by a Modified Polyol Process for Direct Methanol Fuel Cells

    Institute of Scientific and Technical Information of China (English)

    ZHANG Junmin; ZHU Fangfang; ZHANG Kunhua; LIU Weiping; GUAN Weiming

    2012-01-01

    Supported PtRu/C catalysts used in direct methanol fuel cells (DMFCs) were prepared by a new modified polyol method.Transmission electron microscopy (TEM),X-ray diffraction (XRD) and cyclic voltammograms (CVs) were carried out to characterize the morphology,composition and the electrochemical properties of the PtRu/C catalyst.The results revealed that the PtRu nanoparticles with small average particle size (≈2.5 nm),and highly dispersed on the carbon support.The PtRu/C catalyst exhibited high catalytic activity and anti-poisoned performance than that of the JM PtRu/C.It is imply that the modified polyol method is efficient for PtRu/C catalyst preparation.

  18. Nano Si preparation by constant cell voltage electrolysis of FFC-Cambridge Process in molten CaCl2

    Institute of Scientific and Technical Information of China (English)

    Ji; Zhao; Shengmei; Lu; Linyan; Hu; Can; Li

    2013-01-01

    Using FFC-Cambridge Process to prepare Si from SiO2 is a promising method to prepare nanostructured and highly pure silicon for solar cells.However,the method still has many problems unsolved and the controlling effect of the cell voltage on silicon product is not clear.Here we report in this article that nano cluster-like silicon product with purity of 99.95%has been prepared by complete conversion of raw material SiO2,quartz glass plate,using constant cell voltage electrolysis FFC-Cambridge Process.By analysis of XRD,EDS,TEM,HRTEM and ICP-AES as well as the discussion from the thermodynamics calculation,the morphology and components of the product based on the change of cell voltage are clarified.It is clear that pure silicon could be prepared at the cell voltage of 1.7 2.1 V in this reaction system.The silicon material have cluster-like structure which are made of silicon nanoparticles in 20 100 nm size.Interestingly,the cluster-like nano structure of the silicon can be tuned by the used cell voltage.The purity,yield and the energy cost of silicon product prepared at the optimized cell voltage are discussed.The purity of the silicon product could be further improved,hence this method is promising for the preparation of solar grade silicon in future.

  19. In vitro preparation of radionuclides labeled blood cells: Status and requirements; Preparation in vitro des cellules du sang marquees par des radionucleides: statut et recommandations

    Energy Technology Data Exchange (ETDEWEB)

    Couret, I. [Service de medecine nucleaire et radiopharmacie, hopital Lapeyronie, CHU de Montpellier, 34 - Montpellier (France); Desruet, M.D. [Service de medecine nucleaire et radiopharmacie, CHU de Grenoble, 38 - Grenoble (France); Bolot, C. [Service de pharmacie, hospices civils de Lyon, groupement hospitalier Est, 69 - Bron (France); Chassel, M.L. [Service de pharmacie et radiopharmacie, centre hospitalier de Chambery, 73 - Chambery (France); Pellegrin, M. [Inserm U896, CRLC Val-d' Aurelle-Paul-Lamarque, IRCM, universite Montpellier 1, 34 - Montpellier (France)

    2010-11-15

    Labelled blood cells permit nuclear medicine imaging using their physiological behaviours. The radiolabeling must be performed in vitro because of the lack of specific markers and requires several highly technical stages of preparation. Labelled blood cells have not the medication drug status, so that the nuclear physician conducting the nuclear test is fully liable. In most cases, the physician delegates the technical responsibility to radio-pharmacists. Although the status of radiolabelled autologous cells is not legally defined and in the absence of a specific repository, it is essential that their preparation is subject to the requirements of the rules of French Good Manufacturing Practice published by Agence francaise de securite sanitaire des produits de sante (Afssaps). It would be desirable to harmonize the practices of radiolabeling cellular blood components by editing a repository. (authors)

  20. Preparation and influence of performance of anodic catalysts for direct methanol fuel cell

    Institute of Scientific and Technical Information of China (English)

    WANG Zhenbo; YIN Geping; SHI Pengfei

    2007-01-01

    This research aims at increasing the utilization of platinum-ruthenium alloy (Pt-Ru) catalysts and thus lowering the catalyst loading in anodes for methanol electrooxidation.The direct methanol fuel cell's (DMFC) anodic catalysts,Pt-Ru/C,were prepared by chemical reduction with a reducing agent added in two kinds of solutions under different circumstances.The reducing agent was added in hot solution with the protection of inert gases or just air,and in cold solution with inert gases.The catalysts were treated at different temperatures.Their performance was tested by cyclic voltammetry and potentiostatic polarization by utilizing their inherent powder microelectrode in 0.5 mol/L CH3OH and 0.5 mol/L H2SO4 solution.The structures and micro-surface images ofthe catalysts were determined and observed by X-ray diffraction and transmission electron microscopy,respectively.The catalyst prepared in inert gases showed a better catalytic performance for methanol electrooxidation than that prepared in air.It resulted in a more homogeneous distribution of the Pt-Ru alloy in carbon.Its size is small,only about 4.5 nm.The catalytic performance is affected by the order of the reducing agent added.The performance of the catalyst prepared by adding the reductant at constant temperature of the solution is better than that prepared by adding it in the solution at 0℃ and then heating it up to the reducing temperature.The structure of the catalyst was modified,and there was an increase in the conversion of ruthenium into the alloyed state and an increase in particle size with the ascension of heat treatment temperature.In addition,the stability of the catalyst was improved after heat treatment.

  1. Religious competence as cultural competence

    Science.gov (United States)

    2012-01-01

    Definitions of cultural competence often refer to the need to be aware and attentive to the religious and spiritual needs and orientations of patients. However, the institution of psychiatry maintains an ambivalent attitude to the incorporation of religion and spirituality into psychiatric practice. This is despite the fact that many patients, especially those from underserved and underprivileged minority backgrounds, are devotedly religious and find much solace and support in their religiosity. I use the case of mental health of African Americans as an extended example to support the argument that psychiatric services must become more closely attuned to religious matters. I suggest ways in which this can be achieved. Attention to religion can aid in the development of culturally competent and accessible services, which in turn, may increase engagement and service satisfaction among religious populations. PMID:22421686

  2. Research on Preparation of Human Immune Cell in vitro with Response to Shrimp Allergen

    Directory of Open Access Journals (Sweden)

    W.B. Pan

    2015-01-01

    Full Text Available Shrimp is one of the most important food allergens. Tropomyosin is its major allergen. Wherein Pen a 1, contains five antibody binding regions, has been identified as the only major shrimp allergen. However, the study on IgE with response to shrimp allergen is still a serious lack, compared with the allergenic proteins. Particularly in the aspects of the preparation of IgE in vitro, it is restricted and can only obtain the complete IgE molecules by polyclonal or monoclonal technology. As for the preparation of small molecule IgE to the shrimp allergen has not yet been reported. This study attempts to carry out research on obtaining of cell materials that are used to clone. It sets up a convenient and efficient immune system in vitro which combines dendritic cell differentiation, allergens immune, mixed lymphocyte culture and so on. Finally the system successfully activates the proliferation of specific B cells and the secretion of a large number of specific IgE antibodies to shrimp allergen.

  3. Differentiating nursing leadership and management competencies.

    Science.gov (United States)

    Jennings, Bonnie Mowinski; Scalzi, Cynthia C; Rodgers, James D; Keane, Anne

    2007-01-01

    As the foundation of evaluating content for nursing leadership and administration courses, leadership and management competencies were identified from a literature review of 140 articles published between 2000-2004. Similarities and differences among the competencies were assessed. A large intersection of common competencies was discovered, indicating a lack of discrimination between leadership and management competencies. Arguably, this fusion ignores the different purposes served by leadership and management. Alternately, the convergence of leadership and management competencies might reflect that traditional distinctions have narrowed. Nevertheless, ambiguity persists regarding essential leadership and management competencies and the way they are reflected in nursing curricula. If concerns about the work environment are to be remedied for patients and staff, nurse educators are urged to act quickly to identify requisite competencies and better align course content with them. This will better prepare nurse administrators to succeed in their arduous work.

  4. Reducing time and increasing sensitivity in sample preparation for adherent mammalian cell metabolomics.

    Science.gov (United States)

    Lorenz, Matthew A; Burant, Charles F; Kennedy, Robert T

    2011-05-01

    A simple, fast, and reproducible sample preparation procedure was developed for relative quantification of metabolites in adherent mammalian cells using the clonal β-cell line INS-1 as a model sample. The method was developed by evaluating the effect of different sample preparation procedures on high performance liquid chromatography- mass spectrometry quantification of 27 metabolites involved in glycolysis and the tricarboxylic acid cycle on a directed basis as well as for all detectable chromatographic features on an undirected basis. We demonstrate that a rapid water rinse step prior to quenching of metabolism reduces components that suppress electrospray ionization thereby increasing signal for 26 of 27 targeted metabolites and increasing total number of detected features from 237 to 452 with no detectable change of metabolite content. A novel quenching technique is employed which involves addition of liquid nitrogen directly to the culture dish and allows for samples to be stored at -80 °C for at least 7 d before extraction. Separation of quenching and extraction steps provides the benefit of increased experimental convenience and sample stability while maintaining metabolite content similar to techniques that employ simultaneous quenching and extraction with cold organic solvent. The extraction solvent 9:1 methanol: chloroform was found to provide superior performance over acetonitrile, ethanol, and methanol with respect to metabolite recovery and extract stability. Maximal recovery was achieved using a single rapid (∼1 min) extraction step. The utility of this rapid preparation method (∼5 min) was demonstrated through precise metabolite measurements (11% average relative standard deviation without internal standards) associated with step changes in glucose concentration that evoke insulin secretion in the clonal β-cell line INS-1.

  5. PREPARATION AND PROPERTIES OF SPAES-TiO_2 HYBRID MEMBRANES FOR DIRECT METHANOL FUEL CELL

    Institute of Scientific and Technical Information of China (English)

    刘惠玲

    2009-01-01

    Sulfonated poly(arylene ether sulfone)(SPAES) copolymer with degree of sulfonation of 1.0 was synthesized and characterized.A series of SPAES-TiO_2 hybrid membranes with various contents of nano-sized TiO_2 particles were prepared and characterized through sol-gel reactions.Scanning electron microscopy(SEM) images indicated the TiO_2 particles were well dispersed within polymer matrix.These composite membranes were evaluated for proton exchange membranes(PEMs) in direct methanol fuel cell(DMFC).These mem...

  6. Developing the Intercultural Competence of Graduate Students

    Science.gov (United States)

    Dimitrov, Nanda; Dawson, Debra L.; Olsen, Karyn C.; Meadows, Ken N.

    2014-01-01

    This study explores how teaching development programs may facilitate the development of intercultural competence in graduate students and prepare them for communicating effectively in the global workplace after graduation. First, we describe the concept of intercultural teaching competence and examine the skills that graduate students may need to…

  7. Cultivation of corneal endothelial cells on a pericellular matrix prepared from human decidua-derived mesenchymal cells.

    Directory of Open Access Journals (Sweden)

    Ryohei Numata

    Full Text Available The barrier and pump functions of the corneal endothelium are essential for the maintenance of corneal transparency. Although corneal transplantation is the only current therapy for treating corneal endothelial dysfunction, the potential of tissue-engineering techniques to provide highly efficient and less invasive therapy in comparison to corneal transplantation has been highly anticipated. However, culturing human corneal endothelial cells (HCECs is technically difficult, and there is no established culture protocol. The aim of this study was to investigate the feasibility of using a pericellular matrix prepared from human decidua-derived mesenchymal cells (PCM-DM as an animal-free substrate for HCEC culture for future clinical applications. PCM-DM enhanced the adhesion of monkey CECs (MCECs via integrin, promoted cell proliferation, and suppressed apoptosis. The HCECs cultured on the PCM-DM showed a hexagonal morphology and a staining profile characteristic of Na⁺/K⁺-ATPase and ZO-1 at the plasma membrane in vivo, whereas the control HCECs showed a fibroblastic phenotype. The cell density of the cultured HCECs on the PCM-DM was significantly higher than that of the control cells. These results indicate that PCM-DM provides a feasible xeno-free matrix substrate and that it offers a viable in vitro expansion protocol for HCECs while maintaining cellular functions for use as a subsequent clinical intervention for tissue-engineered based therapy of corneal endothelial dysfunction.

  8. Cultivation of corneal endothelial cells on a pericellular matrix prepared from human decidua-derived mesenchymal cells.

    Science.gov (United States)

    Numata, Ryohei; Okumura, Naoki; Nakahara, Makiko; Ueno, Morio; Kinoshita, Shigeru; Kanematsu, Daisuke; Kanemura, Yonehiro; Sasai, Yoshiki; Koizumi, Noriko

    2014-01-01

    The barrier and pump functions of the corneal endothelium are essential for the maintenance of corneal transparency. Although corneal transplantation is the only current therapy for treating corneal endothelial dysfunction, the potential of tissue-engineering techniques to provide highly efficient and less invasive therapy in comparison to corneal transplantation has been highly anticipated. However, culturing human corneal endothelial cells (HCECs) is technically difficult, and there is no established culture protocol. The aim of this study was to investigate the feasibility of using a pericellular matrix prepared from human decidua-derived mesenchymal cells (PCM-DM) as an animal-free substrate for HCEC culture for future clinical applications. PCM-DM enhanced the adhesion of monkey CECs (MCECs) via integrin, promoted cell proliferation, and suppressed apoptosis. The HCECs cultured on the PCM-DM showed a hexagonal morphology and a staining profile characteristic of Na⁺/K⁺-ATPase and ZO-1 at the plasma membrane in vivo, whereas the control HCECs showed a fibroblastic phenotype. The cell density of the cultured HCECs on the PCM-DM was significantly higher than that of the control cells. These results indicate that PCM-DM provides a feasible xeno-free matrix substrate and that it offers a viable in vitro expansion protocol for HCECs while maintaining cellular functions for use as a subsequent clinical intervention for tissue-engineered based therapy of corneal endothelial dysfunction.

  9. Upregulated expression of indoleamine 2, 3-dioxygenase in CHO cells induces apoptosis of competent T cells and increases proportion of Treg cells

    Directory of Open Access Journals (Sweden)

    Liu Juntian

    2011-09-01

    Full Text Available Abstract Introduction The inflammatory enzyme indoleamine 2, 3-dioxygenase (IDO participates in immune tolerance and promotes immune escape of IDO+ tumors. A recent hypothesis suggested that IDO may contribute to the differentiation of new T regulatory cells (Tregs from naive CD4+ T cells. In this study we investigated the role of IDO in induction of immunosuppression in breast cancer by increasing the apoptosis of T cells and the proportion of Tregs. Methods An IDO expression plasmid was constructed and Chinese hamster ovary (CHO cells were stably transfected with human IDO. Purified CD3+ T cells were isolated from the peripheral blood monouclear cells of breast cancer patients. After co-culturing IDO expressing or untransfected (control CHO cells with T cells, T cells apoptosis were determined by flow cytometry analysis and annexin-V and PI staining. The proportion of the regulatory T cell (Tregs [CD4 + CD25 + CD127-] subset was measured by flow cytometry analysis. T cells total RNA and cellular protein samples were isolated for detecting Foxp3 gene and protein expression. Results IDO transgenic CHO cells yielded high levels of IDO enzymatic activity, resulting in complete depletion of tryptophan from the culture medium. We found that apoptosis occurred in 79.07 ± 8.13% of CD3+T cells after co-cultured with IDO+ CHO cells for 3 days and the proportion of CD4 + CD25 + CD127- T cells increased from 3.43 ± 1.07% to 8.98 ± 1.88% (P Conclusions These results suggest that IDO helps to create a tolerogenic milieu in breast tumors by directly inducing T cell apoptosis and enhancing Treg-mediated immunosuppression.

  10. Upregulated expression of indoleamine 2, 3-dioxygenase in CHO cells induces apoptosis of competent T cells and increases proportion of Treg cells.

    Science.gov (United States)

    Sun, Jingyan; Yu, Jinpu; Li, Hui; Yang, Lili; Wei, Feng; Yu, Wenwen; Liu, Juntian; Ren, Xiubao

    2011-09-14

    The inflammatory enzyme indoleamine 2, 3-dioxygenase (IDO) participates in immune tolerance and promotes immune escape of IDO+ tumors. A recent hypothesis suggested that IDO may contribute to the differentiation of new T regulatory cells (Tregs) from naive CD4+ T cells. In this study we investigated the role of IDO in induction of immunosuppression in breast cancer by increasing the apoptosis of T cells and the proportion of Tregs. An IDO expression plasmid was constructed and Chinese hamster ovary (CHO) cells were stably transfected with human IDO. Purified CD3+ T cells were isolated from the peripheral blood monouclear cells of breast cancer patients. After co-culturing IDO expressing or untransfected (control) CHO cells with T cells, T cells apoptosis were determined by flow cytometry analysis and annexin-V and PI staining. The proportion of the regulatory T cell (Tregs [CD4 + CD25 + CD127⁻]) subset was measured by flow cytometry analysis. T cells total RNA and cellular protein samples were isolated for detecting Foxp3 gene and protein expression. IDO transgenic CHO cells yielded high levels of IDO enzymatic activity, resulting in complete depletion of tryptophan from the culture medium. We found that apoptosis occurred in 79.07 ± 8.13% of CD3+T cells after co-cultured with IDO+ CHO cells for 3 days and the proportion of CD4 + CD25 + CD127⁻ T cells increased from 3.43 ± 1.07% to 8.98 ± 1.88% (P Tregs in vitro. Increased expression of Foxp3, a key molecular marker of Tregs, was confirmed by RT-PCR, real-time RT-PCR and Western blot analysis at the same time. These results suggest that IDO helps to create a tolerogenic milieu in breast tumors by directly inducing T cell apoptosis and enhancing Treg-mediated immunosuppression.

  11. Preparation of Nanoporous TiO2 Electrodes for Dye-Sensitized Solar Cells

    Directory of Open Access Journals (Sweden)

    Hsiue-Hsyan Wang

    2011-01-01

    Full Text Available Nano-porous TiO2 thin films have been widely used as the working electrodes in dye-sensitized solar cells (DSSCs. In this work, the phase-pure anatase TiO2 (a-TiO2 and rutile TiO2 (r-TiO2 have been prepared using hydrothermal processes. The investigation of photo-to-electron conversion efficiency of DSSCs fabricated from mixed-TiO2 with a-TiO2 and r-TiO2 ratio of 80 : 20 (A8R2 was performed and compared to that from commercial TiO2 (DP-25. The results showed higher efficiency of DSSC for A8R2 cells with same dependence of cell efficiency on the film thickness for both A8R2 and DP-25 cells. The best efficiency obtained in this work is 5.2% from A8R2 cell with TiO2 film thickness of 12.0 μm. The correlation between the TiO2 films thickness and photoelectron chemical properties of DSSCs fabricated from A8R2 and DP-25 was compared and discussed.

  12. Preparation of anatase/rutile mixed-phase titania nanoparticles for dye-sensitized solar cells.

    Science.gov (United States)

    Hwang, Yong-Kyung; Park, Sung Soo; Lim, Jun-Heok; Won, Yong Sun; Huh, Seong

    2013-03-01

    Acid-labile high surface mesoporous ZnO/Zn(OH)2 composite material is used as a novel hard template for the preparation of mesoporous amorphous TiO2. The template-free amorphous TiO2 material is then thermally crystallized at suitable temperature to control the relative ratio of anatase and rutile phases in a particle. Four different anatase/rutile (AR) mixed-phase TiO2 nanoparticles (AR-3, AR-15, AR-20, and AR-23 denoted for the samples of 3%, 15%, 20%, and 23% rutile phase, respectively) are prepared and characterized by powder X-ray diffraction (PXRD) and transmission electron microscopy (TEM). The coexistence of anatase and rutile phases in a TiO2 nanoparticle is visually confirmed by HRTEM analysis. These mixed-phase TiO2 nanoparticles are examined as candidates for photoelectrodes of dye-sensitized solar cells (DSSCs). The J-V curves and IPCE spectra for the DSSCs prepared from the mixed-phase TiO2 nanoparticles are obtained, and their photovoltaic properties are investigated. The photo-conversion efficiency (eta) indicates the highest value of 5.07% for AR-20. The synergistic effect of coexisting anatase and rutile phases with an optimal ratio in a TiO2 nanoparticle of AR-20 for an efficient interfacial transfer of photo-generated electrons is likely to lead to the highest efficiency among the AR-n samples.

  13. Dye-Sensitized Solar Cells with Anatase TiO2 Nanorods Prepared by Hydrothermal Method

    Directory of Open Access Journals (Sweden)

    Ming-Jer Jeng

    2013-01-01

    Full Text Available The hydrothermal method provides an effective reaction environment for the synthesis of nanocrystalline materials with high purity and well-controlled crystallinity. In this work, we started with various sizes of commercial TiO2 powders and used the hydrothermal method to prepare TiO2 thin films. We found that the synthesized TiO2 nanorods were thin and long when smaller TiO2 particles were used, while larger TiO2 particles produced thicker and shorter nanorods. We also found that TiO2 films prepared by TiO2 nanorods exhibited larger surface roughness than those prepared by the commercial TiO2 particles. It was found that a pure anatase phase of TiO2 nanorods can be obtained from the hydrothermal method. The dye-sensitized solar cells fabricated with TiO2 nanorods exhibited a higher solar efficiency than those fabricated with commercial TiO2 nanoparticles directly. Further, triple-layer structures of TiO2 thin films with different particle sizes were investigated to improve the solar efficiency.

  14. Magnetic Field-Assisted Perovskite Film Preparation for Enhanced Performance of Solar Cells.

    Science.gov (United States)

    Wang, Haoxu; Lei, Jie; Gao, Fei; Yang, Zhou; Yang, Dong; Jiang, Jiexuan; Li, Juan; Hu, Xihong; Ren, Xiaodong; Liu, Bin; Liu, Jing; Lei, Hairui; Liu, Zhike; Liu, Shengzhong Frank

    2017-07-05

    Perovskite solar cells (PSCs) are promising low-cost photovoltaic technologies with high power conversion efficiency (PCE). The crystalline quality of perovskite materials is crucial to the photovoltaic performance of the PSCs. Herein, a simple approach is introduced to prepare high-quality CH3NH3PbI3 perovskite films with larger crystalline grains and longer carriers lifetime by using magnetic field to control the nucleation and crystal growth. The fabricated planar CH3NH3PbI3 solar cells have an average PCE of 17.84% and the highest PCE of 18.56% using an optimized magnetic field at 80 mT. In contrast, the PSCs fabricated without the magnetic field give an average PCE of 15.52% and the highest PCE of 16.72%. The magnetic field action produces an ordered arrangement of the perovskite ions, improving the crystallinity of the perovskite films and resulting in a higher PCE.

  15. Carbons prepared from coals for anodes of lithium-ion cells

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, T.; Xing, W.; Dahn, J.R. [Simon Fraser University, Burnaby, BC (Canada). Dept. of Physics

    1996-12-31

    Carbons were prepared by pyrolysing eight different coals from the Argonne Premium coal sample bank at temperatures near 1000{degree}C. Electrochemical cells were made to study lithium insertion in these carbons. The electrochemical behavior and physical properties of these pyrolysed coals was compared to that of pyrolysed sugar, PVC and pitch. Using powder X-ray diffraction and small angle X-ray scattering (SAXS), it was shown that pyrolysed coals which have the smallest fraction of parallel-stacked graphene layers and hence the largest number of nanoscopic pores per unit mass have the largest capacity for lithium insertion. A reversible capacity of about 450 mAh/g was attained for lithium/carbon cells made from Blind Canyon Seam coal. 17 refs., 10 figs., 3 tabs.

  16. Preparation of Desirable Porous Cell Structure Polylactide/Wood Flour Composite Foams Assisted by Chain Extender

    Directory of Open Access Journals (Sweden)

    Youyong Wang

    2017-08-01

    Full Text Available Polylactide (PLA/wood flour composite foam were prepared through a batch foaming process. The effect of the chain extender on the crystallization behavior and dynamic rheological properties of the PLA/wood flour composites were investigated as well as the crystal structure and cell morphology of the composite foams. The incorporation of the chain extender enhanced the complex viscosity and storage modulus of PLA/wood flour composites, indicating the improved melt elasticity. The chain extender also led to a decreased crystallization rate and final crystallinity of PLA/wood flour composites. With an increasing chain extender content, a finer and more uniform cell structure was formed, and the expansion ratio of PLA/wood flour composite foams was much higher than without the chain extender. Compared to the unfoamed composites, the crystallinity of the foamed PLA/wood flour composites was improved and the crystal was loosely packed. However, the new crystalline form was not evident.

  17. Unconventional Specimen Preparation Techniques Using High Resolution Low Voltage Field Emission Scanning Electron Microscopy to Study Cell Motility, Host Cell Invasion, and Internal Cell Structures in Toxoplasma gondii

    Science.gov (United States)

    Schatten, Heide; Ris, Hans

    2002-04-01

    Apicomplexan parasites employ complex and unconventional mechanisms for cell locomotion, host cell invasion, and cell division that are only poorly understood. While immunofluorescence and conventional transmission electron microscopy have been used to answer questions about the localization of some cytoskeletal proteins and cell organelles, many questions remain unanswered, partly because new methods are needed to study the complex interactions of cytoskeletal proteins and organelles that play a role in cell locomotion, host cell invasion, and cell division. The choice of fixation and preparation methods has proven critical for the analysis of cytoskeletal proteins because of the rapid turnover of actin filaments and the dense spatial organization of the cytoskeleton and its association with the complex membrane system. Here we introduce new methods to study structural aspects of cytoskeletal motility, host cell invasion, and cell division of Toxoplasma gondii, a most suitable laboratory model that is representative of apicomplexan parasites. The novel approach in our experiments is the use of high resolution low voltage field emission scanning electron microscopy (LVFESEM) combined with two new specimen preparation techniques. The first method uses LVFESEM after membrane extraction and stabilization of the cytoskeleton. This method allows viewing of actin filaments which had not been possible with any other method available so far. The second approach of imaging the parasite's ultrastructure and interactions with host cells uses semithick sections (200 nm) that are resin de-embedded (Ris and Malecki, 1993) and imaged with LVFESEM. This method allows analysis of structural detail in the parasite before and after host cell invasion and interactions with the membrane of the parasitophorous vacuole as well as parasite cell division.

  18. Exploring Feasibility for Application of Luminescent CdTe Quantum Dots Prepared in Aqueous Phase to Live Cell Imaging

    Institute of Scientific and Technical Information of China (English)

    Ji Fang WENG; Xing Tao SONG; Liang LI; Hui Feng QIAN; Ke Ying CHEN; Xue Ming XU; Cheng Xi CAO; Ji Cun REN

    2006-01-01

    This paper explored the feasibility for the application of luminescent CdTe quantum dots prepared in aqueous phase to live cell imaging. The highly luminescent CdTe quantum dots (QDs)were first prepared in aqueous phase, and then were covalently coupled to a plant lectin (UEA-1),as a fluorescent probe. After incubating with of human umbilical vein endothelial cells (HUVECs), the QD probe with UEA-1 was able to specifically bind the corresponding cell receptor. The good cell images were obtained in live cells using laser confocal scanning microscopy. We predict that QDs prepared in water phase will probably become an attractive alternative probe in cellular imaging and bio-labeling.

  19. Quantum dot sensitized solar cells with improved efficiency prepared using electrophoretic deposition.

    Science.gov (United States)

    Salant, Asaf; Shalom, Menny; Hod, Idan; Faust, Adam; Zaban, Arie; Banin, Uri

    2010-10-26

    Quantum dot sensitized solar cells (QDSSC) may benefit from the ability to tune the quantum dot optical properties and band gap through the manipulation of their size and composition. Moreover, the inorganic nanocrystals may provide increased stability compared to organic sensitizers. We report the facile fabrication of QDSSC by electrophoretic deposition of CdSe QDs onto conducting electrodes coated with mesoporous TiO(2). Unlike prior chemical linker-based methods, no pretreatment of the TiO(2) was needed, and deposition times as short as 2 h were sufficient for effective coating. Cross-sectional chemical analysis shows that the Cd content is nearly constant across the entire TiO(2) layer. The dependence of the deposition on size was studied and successfully applied to CdSe dots with diameters between 2.5 and 5.5 nm as well as larger CdSe quantum rods. The photovoltaic characteristics of the devices are greatly improved compared with those achieved for cells prepared with a linker approach, reaching efficiencies as high as 1.7%, under 1 sun illumination conditions, after treating the coated electrodes with ZnS. Notably, the absorbed photon to electron conversion efficiencies did not show a clear size-dependence indicating efficient electron injection even for the larger QD sizes. The electrophoretic deposition method can be easily expanded and applied for preparations of QDSSCs using diverse colloidal quantum dot and quantum rod materials for sensitization.

  20. Characterization and single cell testing of Pt/C electrodes prepared by electrodeposition

    Energy Technology Data Exchange (ETDEWEB)

    Martin, A.J.; Chaparro, A.M.; Gallardo, B.; Folgado, M.A. [CIEMAT, Department of Energy, Avda. Complutense 22, 28040 Madrid (Spain); Daza, L. [CIEMAT, Department of Energy, Avda. Complutense 22, 28040 Madrid (Spain); Instituto de Catalisis y Petroleoquimica (CSIC), C/. Marie Curie 2, Campus Cantoblanco, 28049 Madrid (Spain)

    2009-07-01

    Electrodes for proton exchange membrane fuel cells (PEMFC) have been prepared by the electrodeposition method. For this task, the electrodeposition of platinum is carried out on a carbon black substrate impregnated with an ionomer, proton conducting, medium. Before electrodeposition, the substrate is submitted to an activation process to increase the hydrophilic character of the surface to a few microns depth. Electrodeposition of platinum takes place inside the generated surface hydrophilic layer, resulting in a continuous phase covering totally or partially carbon substrate grains. Cross sectional images show a decay profile of platinum towards the interior of the substrate, reflecting a deposition process limited by diffusion of PtCl{sub 6}{sup 2-} through the porous substrate. Electrodes with different platinum loads have been prepared, and membrane electrode assemblies (MEA) have been mounted with the electrodeposited electrodes as cathode and other standard components (commercial anode and Nafion{sup R} 117 membrane). The electrochemically active surface area determined from hydrogen underpotential deposition charge, is lower on the electrodeposited electrodes than on standard electrodes. However, single cell testing shows higher mass specific activity on electrodeposited cathodes with low and intermediate Pt load (below 0.05 mg Pt cm{sup -2}). (author)

  1. Characterization and single cell testing of Pt/C electrodes prepared by electrodeposition

    Science.gov (United States)

    Martín, A. J.; Chaparro, A. M.; Gallardo, B.; Folgado, M. A.; Daza, L.

    Electrodes for proton exchange membrane fuel cells (PEMFC) have been prepared by the electrodeposition method. For this task, the electrodeposition of platinum is carried out on a carbon black substrate impregnated with an ionomer, proton conducting, medium. Before electrodeposition, the substrate is submitted to an activation process to increase the hydrophilic character of the surface to a few microns depth. Electrodeposition of platinum takes place inside the generated surface hydrophilic layer, resulting in a continuous phase covering totally or partially carbon substrate grains. Cross sectional images show a decay profile of platinum towards the interior of the substrate, reflecting a deposition process limited by diffusion of PtCl 6 2- through the porous substrate. Electrodes with different platinum loads have been prepared, and membrane electrode assemblies (MEA) have been mounted with the electrodeposited electrodes as cathode and other standard components (commercial anode and Nafion R 117 membrane). The electrochemically active surface area determined from hydrogen underpotential deposition charge, is lower on the electrodeposited electrodes than on standard electrodes. However, single cell testing shows higher mass specific activity on electrodeposited cathodes with low and intermediate Pt load (below 0.05 mg Pt cm -2).

  2. Preparation of active layers in polymer solar cells by aerosol jet printing.

    Science.gov (United States)

    Yang, Chunhe; Zhou, Erjun; Miyanishi, Shoji; Hashimoto, Kazuhito; Tajima, Keisuke

    2011-10-01

    Active layers of polymer solar cells were prepared by aerosol jet printing of organic inks. Various solvents and additives with high boiling points were screened for the preparation of high-quality polymer films. The effects on device performance of treating the films by thermal and solvent vapor annealing were also investigated. The components of the solvent were important for controlling the drying rate of the liquid films, reducing the number of particle-like protrusions on the film surface, and realizing high molecular ordering in the polymer phases. The optimized solar cell device with poly(3-hexylthiophene) and a C(60) derivative showed a high fill factor of 67% and power conversion efficiency of 2.53% without thermal annealing. The combination of poly[N-9-heptadecanyl-2,7-carbazole-alt-3,6-bis(thiophen-5-yl)-2,5-diethylhexyl-2,5-dihydropyrrolo-[3,4-]pyrrole-1,4-dione] and a C(70) derivative led to power conversion efficiency of 3.92 and 3.14% for device areas of 0.03 and 1 cm(2), respectively.

  3. Preparation, cell compatibility and degradability of collagen-modified poly(lactic acid).

    Science.gov (United States)

    Cui, Miaomiao; Liu, Leili; Guo, Ning; Su, Ruixia; Ma, Feng

    2015-01-05

    Poly(lactic acid) (PLA) was modified using collagen through a grafting method to improve its biocompatibility and degradability. The carboxylic group at the open end of PLA was transferred into the reactive acylchlorided group by a reaction with phosphorus pentachloride. Then, collagen-modified PLA (collagen-PLA) was prepared by the reaction between the reactive acylchlorided group and amino/hydroxyl groups on collagen. Subsequently, the structure of collagen-PLA was confirmed by Fourier transform infrared spectroscopy, fluorescein isothiocyanate-labeled fluorescence spectroscopy, X-ray photoelectron spectroscopy, and DSC analyses. Finally, some properties of collagen-PLA, such as hydrophilicity, cell compatibility and degradability were characterized. Results showed that collagen had been grafted onto the PLA with 5% graft ratio. Water contact angle and water absorption behavior tests indicated that the hydrophilicity of collagen-PLA was significantly higher than that of PLA. The cell compatibility of collagen-PLA with mouse embryonic fibroblasts (3T3) was also significantly better than PLA in terms of cell morphology and cell proliferation, and the degradability of PLA was also improved after introducing collagen. Results suggested that collagen-PLA was a promising candidate for biomedical applications.

  4. Low-cost plasmonic solar cells prepared by chemical spray pyrolysis.

    Science.gov (United States)

    Kärber, Erki; Katerski, Atanas; Oja Acik, Ilona; Mikli, Valdek; Mere, Arvo; Sildos, Ilmo; Krunks, Malle

    2014-01-01

    Solar cells consisting of an extremely thin In2S3/CuInS2 buffer/absorber layer uniformly covering planar ZnO were prepared entirely by chemical spray pyrolysis. Au nanoparticles (Au-NPs) were formed via thermal decomposition of a gold(III) chloride trihydrate (HAuCl4·3H2O) precursor by spraying 2 mmol/L of the aqueous precursor solution onto a substrate held at 260 °C. Current-voltage scans and external quantum efficiency spectra were used to evaluate the solar cell performance. This work investigates the effect of the location of the Au-NP layer deposition (front side vs rear side) in the solar cell and the effect of varying the volume (2.5-10 mL) of the sprayed Au precursor solution. A 63% increase (from 4.6 to 7.5 mA/cm(2)) of the short-circuit current density was observed when 2.5 mL of the precursor solution was deposited onto the rear side of the solar cell.

  5. Preparation of nano/submicrometer yam and its benefits on collagen secretion from skin fibroblast cells.

    Science.gov (United States)

    Chiang, Lung-Hsuan; Chen, Shih-Hsin; Yeh, An-I

    2012-12-19

    Nano/submicrometer-scaled yam particles have been prepared by using media-milling. The particle size of media-milled yam was confirmed by the laser light scattering method and scanning electron microscopy. Influences of media-milled yam on skin fibroblast cells (WS1) were evaluated. The size reduction did not significantly alter the proximate composition, and the presence of nanoparticles was not toxic to WS1 cells. The contents of bioactive compounds (diosgenin, stigmasterol, and β-sitosterol) were significantly increased by media-milling, which enhanced the secretion of hTGF-β and inhibited the formation of MMP-1. Thus, the collagen secretion from WS1 was significantly increased by size reduction. Diosgenin was employed as a positive control. Nevertheless, media-milled yam exhibited greater effects on WS1 cells than diosgenin. It appeared that both diosgenin and size reduction were helpful for enhancing the secretion of collagen by WS1 cells. In addition, the irritancy of yam was eliminated by media-milling.

  6. Low-cost plasmonic solar cells prepared by chemical spray pyrolysis

    Directory of Open Access Journals (Sweden)

    Erki Kärber

    2014-12-01

    Full Text Available Solar cells consisting of an extremely thin In2S3/CuInS2 buffer/absorber layer uniformly covering planar ZnO were prepared entirely by chemical spray pyrolysis. Au nanoparticles (Au-NPs were formed via thermal decomposition of a gold(III chloride trihydrate (HAuCl4·3H2O precursor by spraying 2 mmol/L of the aqueous precursor solution onto a substrate held at 260 °C. Current–voltage scans and external quantum efficiency spectra were used to evaluate the solar cell performance. This work investigates the effect of the location of the Au-NP layer deposition (front side vs rear side in the solar cell and the effect of varying the volume (2.5–10 mL of the sprayed Au precursor solution. A 63% increase (from 4.6 to 7.5 mA/cm2 of the short-circuit current density was observed when 2.5 mL of the precursor solution was deposited onto the rear side of the solar cell.

  7. The Preparation of Bioimprinted Whole-cell Biocatalyst and Its Application in Bioconversion of Biodiesel

    Directory of Open Access Journals (Sweden)

    Meiling Chen

    2015-08-01

    Full Text Available Biodiesel has attracted considerable attention as an environmentally friendly alternative fuel. Lipase is the most popular enzyme for biodiesel production and immobilization has been deployed to improve enzyme stability and reusability. Exploitation of high activity lipase is the key point for biodiesel production. Whole-cell biocatalysts have been applied in the biosynthesis of biodiesel and bioimprinting is a promising approach for enzyme performances improvement. In this study, based on the S. cerevisiae cell-surface display system with &alpha-agglutinin as anchor, a whole-cell biocatalyst of codon-optimized Rhizopus oryzae lipase was constructed and bioimprinted with oleic acid, gaining 5-fold increase on enzymatic activity in the alcoholysis of soybean oil to biodiesel. Moreover, the conversion of FAME was up to 95.45±2.73% after a 27-h reaction at 60°C. Our results indicated that combining bioimprinting with yeast display technique to prepare whole-cell biocatalyst could result in potential enzymes for bioconversion of biodiesel in organic solvents.

  8. Low-cost plasmonic solar cells prepared by chemical spray pyrolysis

    Science.gov (United States)

    Katerski, Atanas; Oja Acik, Ilona; Mikli, Valdek; Mere, Arvo; Sildos, Ilmo; Krunks, Malle

    2014-01-01

    Summary Solar cells consisting of an extremely thin In2S3/CuInS2 buffer/absorber layer uniformly covering planar ZnO were prepared entirely by chemical spray pyrolysis. Au nanoparticles (Au-NPs) were formed via thermal decomposition of a gold(III) chloride trihydrate (HAuCl4·3H2O) precursor by spraying 2 mmol/L of the aqueous precursor solution onto a substrate held at 260 °C. Current–voltage scans and external quantum efficiency spectra were used to evaluate the solar cell performance. This work investigates the effect of the location of the Au-NP layer deposition (front side vs rear side) in the solar cell and the effect of varying the volume (2.5–10 mL) of the sprayed Au precursor solution. A 63% increase (from 4.6 to 7.5 mA/cm2) of the short-circuit current density was observed when 2.5 mL of the precursor solution was deposited onto the rear side of the solar cell. PMID:25551068

  9. Cell-laden microgel prepared using a biocompatible aqueous two-phase strategy.

    Science.gov (United States)

    Liu, Yang; Nambu, Natalia Oshima; Taya, Masahito

    2017-09-01

    Microfluidic methods are frequently used to produce cell-laden microgels for various biomedical purposes. Such microfluidic methods generally employ oil-water systems. The poor distribution of crosslinking reagents in the oil phase limits the available gelation strategies. Extracting the microgel from the oil-phase also reduces its production efficiency. In this study, an aqueous two-phase system (ATPS) involving dextran (DEX) and polyethylene glycol (PEG) was used to prepare cell-laden microgel. This avoided the problems associated with an oil phase. The microgel precursor polymers and crosslinking reagents were dispersed in the DEX and PEG phases, respectively. The ultra-low interfacial tension of the ATPS hindered droplet formation. A co-flow microfluidic device was fabricated to overcome this problem. The device incorporated a square-wave-changing injection force, to improve the efficiency of droplet formation. The microgel precursor (including alginate and carboxymethyl cellulose derivatives possessing phenolic hydroxyl moieties) could be dispersed in the DEX solution at various concentrations. Uniform droplets were formed with controllable diameters, and were sequentially converted to microgel by horseradish peroxidase-catalyzed crosslinking. Cells were dispersed in the DEX phase with the microgel precursor polymer, and retained their high viability and proliferation in the resulting microgel. The solubility of gelatin derivatives in the DEX phase was low, but was sufficient to impart cell adhesion properties on the microgel.

  10. Type I interferon reaction to viral infection in interferon-competent, immortalized cell lines from the African fruit bat Eidolon helvum.

    Directory of Open Access Journals (Sweden)

    Susanne E Biesold

    Full Text Available Bats harbor several highly pathogenic zoonotic viruses including Rabies, Marburg, and henipaviruses, without overt clinical symptoms in the animals. It has been suspected that bats might have evolved particularly effective mechanisms to suppress viral replication. Here, we investigated interferon (IFN response, -induction, -secretion and -signaling in epithelial-like cells of the relevant and abundant African fruit bat species, Eidolon helvum (E. helvum. Immortalized cell lines were generated; their potential to induce and react on IFN was confirmed, and biological assays were adapted to application in bat cell cultures, enabling comparison of landmark IFN properties with that of common mammalian cell lines. E. helvum cells were fully capable of reacting to viral and artificial IFN stimuli. E. helvum cells showed highest IFN mRNA induction, highly productive IFN protein secretion, and evidence of efficient IFN stimulated gene induction. In an Alphavirus infection model, O'nyong-nyong virus exhibited strong IFN induction but evaded the IFN response by translational rather than transcriptional shutoff, similar to other Alphavirus infections. These novel IFN-competent cell lines will allow comparative research on zoonotic, bat-borne viruses in order to model mechanisms of viral maintenance and emergence in bat reservoirs.

  11. Parvovirus B19 promoter at map unit 6 confers autonomous replication competence and erythroid specificity to adeno-associated virus 2 in primary human hematopoietic progenitor cells.

    Science.gov (United States)

    Wang, X S; Yoder, M C; Zhou, S Z; Srivastava, A

    1995-01-01

    The pathogenic human parvovirus B19 is an autonomously replicating virus with a remarkable tropism for human erythroid progenitor cells. Although the target cell specificity for B19 infection has been suggested to be mediated by the erythrocyte P-antigen receptor (globoside), a number of nonerythroid cells that express this receptor are nonpermissive for B19 replication. To directly test the role of expression from the B19 promoter at map unit 6 (B19p6) in the erythroid cell specificity of B19, we constructed a recombinant adeno-associated virus 2 (AAV), in which the authentic AAV promoter at map unit 5 (AAVp5) was replaced by the B19p6 promoter. Although the wild-type (wt) AAV requires a helper virus for its optimal replication, we hypothesized that inserting the B19p6 promoter in a recombinant AAV would permit autonomous viral replication, but only in erythroid progenitor cells. In this report, we provide evidence that the B19p6 promoter is necessary and sufficient to impart autonomous replication competence and erythroid specificity to AAV in primary human hematopoietic progenitor cells. Thus, expression from the B19p6 promoter plays an important role in post-P-antigen receptor erythroid-cell specificity of parvovirus B19. The AAV-B19 hybrid vector system may also prove to be useful in potential gene therapy of human hemoglobinopathies. Images Fig. 2 Fig. 3 Fig. 4 PMID:8618912

  12. Human amniotic fluid derived cells can competently substitute dermal fibroblasts in a tissue-engineered dermo-epidermal skin analog

    NARCIS (Netherlands)

    Hartmann-Fritsch, Fabienne; Hosper, Nynke; Luginbuehl, Joachim; Biedermann, Thomas; Reichmann, Ernst; Meuli, Martin

    Human amniotic fluid comprises cells with high differentiation capacity, thus representing a potential cell source for skin tissue engineering. In this experimental study, we investigated the ability of human amniotic fluid derived cells to substitute dermal fibroblasts and support epidermis

  13. Modification of competence for in vitro response to Fusarium oxysporum in tomato cells. II. Effect of the integration of Agrobacterium tumefaciens genes for auxin and cytokinin synthesis.

    Science.gov (United States)

    Storti, E; Bogani, P; Bettini, P; Bittini, P; Guardiola, M L; Pellegrini, M G; Inzé, D; Buiatti, M

    1994-04-01

    We have studied the effect of a change in the endogenous hormone equilibria on the competence of tomato (Lycopersicon esculentum) cells to defend themselves against the fungal pathogen Fusarium oxysporum f. sp. lycopersici. Calluses from cvs 'Davis' and 'Red River', respectively resistant and susceptible to Fusarium and transgenic for an auxin- or cytokinin-synthesizing gene from Agrobacterium tumefaciens, were used. The integration of Agrobacterium hormone-related genes into susceptible cv 'Red River' can bring the activation of defense processes to a stable competence as assessed by the inhibition of mycelial growth in dual culture and gem-tube elongation of Fusarium conidia, the determination of callose contents, peroxidase induction and ion leakage in the presence of fusaric acid. This is particularly true when the transformation results in a change of phytohormone equilibria towards an higher cytokin in concentration. On the contrary, in resistant cv 'Davis' the inhibition of both fungal growth in dual culture and conidia germination is higher when the hormone balance is modified in favour of the auxins. No significant effect was observed for ion leakage and peroxidase induction, probably because of a constitutive overproduction of cytokinins in 'Davis' cells.

  14. Application of immobilized cell preparation obtained from biomass of Gluconacetobacter xylinus bacteria in biotransformation of glycerol to dihydroxyacetone

    Directory of Open Access Journals (Sweden)

    Lidia Stasiak-Różańska

    2011-03-01

    Full Text Available Introduction. Dihydroxyacetone (DHA, being a product of glycerol oxidation by acetic acid bacteria, is an important compound widely applied in the cosmetic, food, and pharmaceutical industry, as well as in medicine. Biotransformation of glycerol to DHA is catalyzed by glycerol dehydrogenase (GlyDH, EC 1.1.1.6 bound with the cytoplasmic membrane of bacteria. An attempt was undertaken in this study to conduct glycerol biotransformation with immobilized fractions of a cell preparation with GlyDH activity. The content of dihydroxyacetone obtained with the cell preparation was compared with its content achieved in the reaction with immobilized viable cells of G. xylinus. Material and methods. Cell walls of Gluconacetobacter xylinus bacteria were disintegrated enzymatically. The resultant preparation was immobilized on calcium alginate or first separated into two fractions (precipitate and supernatant by centrifugation and then immobilized. DHA content was determined colorimetrically after the reaction with 3,5-dinitrosalicilic acid. Glycerol content was assayed with the refractometric method. Results. After 20 days of the process, the concentration of DHA obtained with immobilized whole cells reached 25 g/l. In turn, the content of DHA obtained in the same period with immobilized fractions of the cell preparation accounted for 16.9 g/l and 8.95 g/l (depending on the fraction applied. Conclusions. DHA may be obtained in the process independent of G. xylinus metabolic activity using a preparation which displays the catalytic activity of glycerol dehydrogenase and obtained as a result of disintegration of live bacterial cells. The application of such a preparation may in the future eliminate technological problems posed by the presence of bacterial cells and their metabolites in the culture medium.

  15. Generic Engineering Competencies: A Review and Modelling Approach

    Science.gov (United States)

    Male, Sally A.

    2010-01-01

    This paper puts forward the view that engineering educators have a responsibility to prepare graduates for engineering work and careers. The current literature reveals gaps between the competencies required for engineering work and those developed in engineering education. Generic competencies feature in these competency gaps. Literature suggests…

  16. Evaluation of the effects of Streptococcus mutans chaperones and protein secretion machinery components on cell surface protein biogenesis, competence, and mutacin production.

    Science.gov (United States)

    Crowley, P J; Brady, L J

    2016-02-01

    The respective contributions of components of the protein translocation/maturation machinery to cell surface biogenesis in Streptococcus mutans are not fully understood. Here we used a genetic approach to characterize the effects of deletion of genes encoding the ribosome-associated chaperone RopA (Trigger Factor), the surface-localized foldase PrsA, and the membrane-localized chaperone insertases YidC1 and YidC2, both singly and in combination, on bacterial growth, chain length, self-aggregation, cell surface hydrophobicity, autolysis, and antigenicity of surface proteins P1 (AgI/II, PAc), WapA, GbpC, and GtfD. The single and double deletion mutants, as well as additional mutant strains lacking components of the signal recognition particle pathway, were also evaluated for their effects on mutacin production and genetic competence.

  17. Relationship between Nucleus Swelling and Development Competence of Bovine Cloned Embryos Reconstructed by Enucleated Oocytes with Serum-starved or Serum-fed Fetal Somatic Cells

    Directory of Open Access Journals (Sweden)

    Mokhamad Fahrudin

    2015-10-01

    Full Text Available This study was conducted to examine the occurrence of nuclear remodeling (nucleus swelling and its effectson the subsequent in vitro development of bovine embryos reconstructed by serum-starved and serum-fed somaticcells. Results from this study demonstrated that all of the reconstructed embryos that received serum-starved andserum-fed somatic cells exhibited condensed-nuclei. More than 90% of the transferred nuclei exhibited nuclearenvelope breakdown and premature chromatin condensation which clearly distinct from an intact nucleus. Therewas no significant difference on the degree of nucleus swelling in SS-NT embryos or SF-NT embryos, indicatingthat either serum-starved or confluent somatic cell lines could be reprogrammed by the recipient cytoplasmenvironments in similar pattern. Although the fusion rate was not significantly different among the groups, theproportion of SS-NT embryos which developed to the 2- to 4-cell stage (89.7% and to the 8- to 16-cell stage (74.7%was significantly higher than that of SF-NT embryos. Whereas, the proportion of reconstructed embryos thatdeveloped to the morula and blastocyst stages were not significantly different among the groups. Results of thesestudies demonstrate that reconstructed embryos, which received either serum-starved or serum-fed confluentsomatic cells, showed similar developmental competence to the blastocyst stage.Keywords: nuclear transplantation technique-somatic cells-nucleus swelling

  18. Mesenchymal Stem Cells for Regenerative Therapy: Optimization of Cell Preparation Protocols

    Directory of Open Access Journals (Sweden)

    Chiho Ikebe

    2014-01-01

    Full Text Available Administration of bone marrow-derived mesenchymal stem cells (MSCs is an innovative approach for the treatment of a range of diseases that are not curable by current therapies including heart failure. A number of clinical trials have been completed and many others are ongoing; more than 2,000 patients worldwide have been administered with culture-expanded allogeneic or autologous MSCs for the treatment of various diseases, showing feasibility and safety (and some efficacy of this approach. However, protocols for isolation and expansion of donor MSCs vary widely between these trials, which could affect the efficacy of the therapy. It is therefore important to develop international standards of MSC production, which should be evidence-based, regulatory authority-compliant, of good medical practice grade, cost-effective, and clinically practical, so that this innovative approach becomes an established widely adopted treatment. This review article summarizes protocols to isolate and expand bone marrow-derived MSCs in 47 recent clinical trials of MSC-based therapy, which were published after 2007 onwards and provided sufficient methodological information. Identified issues and possible solutions associated with the MSC production methods, including materials and protocols for isolation and expansion, are discussed with reference to relevant experimental evidence with aim of future clinical success of MSC-based therapy.

  19. Clastogenic potential of Ruta graveolens extract and a homeopathic preparation in mouse bone marrow cells.

    Science.gov (United States)

    Preethi, Korengath C; Nair, Cherappally K K; Kuttan, Ramadasan

    2008-01-01

    Ruta graveolens belonging to family Rutaceae has long been traditionally used as a medicinal plant as well as a flavoring agent in food. However, very little data are available on the toxicity of the plant. This report presents evidence on the genotoxic and clastogenic potential of an extract of Ruta graveolens and Ruta 200C, a homeopathic preparation. Various types of chromosomal aberrations were noted in bone marrow cells after treatment. The percentage of aberrated cells in the 400mg/kgb.wt extract administered group was found to be 21% and with 1,000 mg/kg.b.wt it was 31%. The value for the Ruta 200C treated group was also elevated to 23% as compared to the 3%for untreated animals. In addition, bone marrow cells had higher incidence of micronuclei induction when treated with the extract (400 mg and 1,000 mg/kg body weight) and Ruta 200C for 30 days. Administration of the extract (1,000 mg/kg.b.wt) over a period of 30 days also resulted in damage to cellular DNA as evidenced by comet formation where the comet parameters such as percentage DNA in tail, tail length, tail moment of the bone marrow cells were increased several fold over control values. The comet tail moment of the bone marrow cells increased from 4.5 to 50.2 after the extract treatment. Administration of Ruta 200C for 5 consecutive days increased the tail moment to 11.7. These results indicate that Ruta graveolens and Ruta 200C may induce genotoxicity in animals.

  20. Defining Tobacco Regulatory Science Competencies.

    Science.gov (United States)

    Wipfli, Heather L; Berman, Micah; Hanson, Kacey; Kelder, Steven; Solis, Amy; Villanti, Andrea C; Ribeiro, Carla M P; Meissner, Helen I; Anderson, Roger

    2017-02-01

    In 2013, the National Institutes of Health and the Food and Drug Administration funded a network of 14 Tobacco Centers of Regulatory Science (TCORS) with a mission that included research and training. A cross-TCORS Panel was established to define tobacco regulatory science (TRS) competencies to help harmonize and guide their emerging educational programs. The purpose of this paper is to describe the Panel's work to develop core TRS domains and competencies. The Panel developed the list of domains and competencies using a semistructured Delphi method divided into four phases occurring between November 2013 and August 2015. The final proposed list included a total of 51 competencies across six core domains and 28 competencies across five specialized domains. There is a need for continued discussion to establish the utility of the proposed set of competencies for emerging TRS curricula and to identify the best strategies for incorporating these competencies into TRS training programs. Given the field's broad multidisciplinary nature, further experience is needed to refine the core domains that should be covered in TRS training programs versus knowledge obtained in more specialized programs. Regulatory science to inform the regulation of tobacco products is an emerging field. The paper provides an initial list of core and specialized domains and competencies to be used in developing curricula for new and emerging training programs aimed at preparing a new cohort of scientists to conduct critical TRS research. © The Author 2016. Published by Oxford University Press on behalf of the Society for Research on Nicotine and Tobacco. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. MORPHOFUNCTIONAL CHARACTERISTICS OF FIBROBLASTS (MCCOY CELL LINE CULTURED WITH MAGNESIUM PREPARATIONS

    Directory of Open Access Journals (Sweden)

    L. V. Didenko

    2015-01-01

    Full Text Available Aim. To study the effect of magnesium orotate, magnesium/pyridoxine combination and magnesium sulfate on fibroblast morphofunctional characteristics in cell culture of fibroblasts (McCoy line.Material and methods. The study of fibroblasts (McCoy line with the addition of magnesium-containing preparations (magnesium orotate, magnesium/pyridoxine combination, magnesium sulphate to the culture medium was performed using scanning and transmission electron microscopy.Results. When adding into the media magnesium orotate or magnesium/pyridoxine combination, significant changes in morphofunctional state of fibroblasts were noted, which were absent when adding magnesium sulfate. At that fibroblasts significantly promoted synthetic and secretory activity. This was expressed in the formation of amorphous and fibrous components in well-formed cell layers.Conclusion. Stimulation by magnesium ions of the proliferative activity of fibroblasts is shown in the morphological analysis of the effect of magnesium orotate or magnesium in combination with pyridoxine and magnesium sulfate on morphological and functional organization of fibroblasts. Revealed the predominant influence of magnesium orotate, and a less pronounced effect of magnesium in combination with pyridoxine on biosynthetic processes in the cells, including the synthesis of amorphous and fibrous components (protocollagen and elastin of the extracellular matrix founded.

  2. Preparative chromatography of flavonoids and saponins in Gynostemma pentaphyllum and their antiproliferation effect on hepatoma cell.

    Science.gov (United States)

    Tsai, Y C; Lin, C L; Chen, B H

    2010-12-15

    A preparative column chromatographic method was developed to isolate flavonoids and saponins from Gynostemma pentaphyllum, a Chinese Medicinal herb, and evaluate their antiproliferation effect on hepatoma cell Hep3B, with the standards rutin and ginsenoside Rb(3) being used for comparison. Initially the powdered G. pentaphyllum was extracted with ethanol, followed by eluting flavonoids and saponins with ethanol-water (30:70, v/v) and 100% ethanol, respectively, in an open-column containing 5 g of Cosmosil 75C(18)-OPN, and then subjected to HPLC-MS analysis. The flavonoid fraction was mainly composed of quercetin- and kaempferol-glycosides, while in saponin fraction, both ginsenoside Rb(3) and ginsenoside Rd dominated. Both fractions were more effective against Hep3B cells than the standards rutin and ginsenoside Rb(3), with the cell cycle being arrested at G0/G1 phase for all the treatments. Additionally, the inhibition effect followed a dose-dependent increase for all the sample treatments. The result of this study may be used as a basis for possible phytopreparations in the future with G. pentaphyllum as raw material. Copyright © 2010 Elsevier GmbH. All rights reserved.

  3. Preparation, blood coagulation and cell compatibility evaluation of chitosan-graft-polylactide copolymers.

    Science.gov (United States)

    Wang, Qi; Liu, Pei; Liu, Peifeng; Gong, Tao; Li, Suming; Duan, Yourong; Zhang, Zhirong

    2014-02-01

    Biodegradable chitosan-graft-polylactide (PLA-CS) copolymers were prepared by the grafting of a poly(L-lactide) (PLLA) or poly(D-lactide) (PDLA) precursor to the backbone of chitosan using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC ⋅ HCl) and N-hydroxysuccinimide (NHS) as a coupling agent. The blood and cell compatibility of the graft copolymers were investigated in comparison to PLLA and PDLA homopolymers. The coagulation properties of PLA-CS were evaluated by hemolysis, plasma recalcification, dynamic blood clotting and protein absorption assays. PLA-CS copolymers present similar hemolysis ratio and plasma recalcification time as PLA, but slower dynamic blood clotting and lower protein absorption. The cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), agar diffusion and lactate dehydrogenase (LDH) experiments. All the samples presented no effect on the viability to cells. Inflammatory cytokine analysis using sandwich ELISAs revealed that PLA-CS would not stimulate inflammatory activity.

  4. Preparation of chitosan films using different neutralizing solutions to improve endothelial cell compatibility.

    Science.gov (United States)

    He, Qing; Ao, Qiang; Gong, Yandao; Zhang, Xiufang

    2011-12-01

    The development of chitosan-based constructs for application in large-size defects or highly vascularized tissues is still a challenging issue. The poor endothelial cell compatibility of chitosan hinders the colonization of vascular endothelial cells in the chitosan-based constructs, and retards the establishment of a functional microvascular network following implantation. The aim of the present study is to prepare chitosan films with different neutralization methods to improve their endothelial cell compatibility. Chitosan salt films were neutralized with either sodium hydroxide (NaOH) aqueous solution, NaOH ethanol solution, or ethanol solution without NaOH. The physicochemical properties and endothelial cell compatibility of the chitosan films were investigated. Results indicated that neutralization with different solutions affected the surface chemistry, swelling ratio, crystalline conformation, nanotopography, and mechanical properties of the chitosan films. The NaOH ethanol solution-neutralized chitosan film (Chi-NaOH/EtOH film) displayed a nanofiber-dominant surface, while the NaOH aqueous solution-neutralized film (Chi-NaOH/H(2)O film) and the ethanol solution-neutralized film (Chi-EtOH film) displayed nanoparticle-dominant surfaces. Moreover, the Chi-NaOH/EtOH films exhibited a higher stiffness as compared to the Chi-NaOH/H(2)O and Chi-EtOH films. Endothelial cell compatibility of the chitosan films was evaluated with a human microvascular endothelial cell line, HMEC-1. Compared with the Chi-NaOH/H(2)O and Chi-EtOH films, HMECs cultured on the Chi-NaOH/EtOH films fully spread and exhibited significantly higher levels of adhesion and proliferation, with retention of the endothelial phenotype and function. Our findings suggest that the surface nanotopography and mechanical properties contribute to determining the endothelial cell compatibility of chitosan films. The nature of the neutralizing solutions can affect the physicochemical properties and

  5. Preparation of Sulfonated PVA-TMSP Membranes for Direct Methanol Fuel Cell

    Directory of Open Access Journals (Sweden)

    Haryadi

    2012-08-01

    Full Text Available Novel preparation and characterization of sulfonated polyvinyl alcohol (PVA–trimethoxysilyl propanethiol (TMSP membranes for direct methanol fuel cell (DMFC application have been investigated. Preparation of sulfonated PVA-TMSP membrane was conducted by crosslinking steps using sol-gel method and a catalyst of concentrated HCl. TMSP concentrations were varied from 1% to 3%. The gel solution was cast on to the membrane metal plate to obtain membrane sheets. The membrane was then oxidized in H2O2 concentrations of (10-30% to convert the mercapto groups into sulfonate group. Investigations of the cross-linking process and the existence of sulfonate group were conducted by infrared spectroscopy as shown for frequencies at 1140–1200/cm and 1200–1145/cm respectively. The scanning electron microscope–energy dispersive X-rays (SEM–EDX of the membranes indicated that the distribution of silica particles from sol–gel reaction products was uneven due to the fast exchange rate of condensation. The degree of swelling decreased as methanol concentrations increase for sulfonated PVA–TMSP membrane which opposed toward the value of commercial Nafion membrane. The maximum value of ion exchange capacity of the membrane was 1.82 mmol/g whereas the highest proton conductivity was 3.9 x 10-4 S/cm. Therefore it can be concluded that the membrane was a potential candidate for application in DMFC.

  6. The Optimization of Matrix Preparation Process and Performance Testing for Molten Carbonate Fuel Cell

    Directory of Open Access Journals (Sweden)

    Jian Cheng

    2014-01-01

    Full Text Available A key component in the molten carbonate fuel cell (MCFC is electrolyte matrix, which provides both ionic conduction and gas sealing. The aim of this work is to investigate the effects of selected operating conditions on the performance of the matrix preparation. Slurries were prepared to produce matrices by the technique of tape casting. The characteristics of the slurries and matrices were examined by laser particle size analyzer, scanning electron microscopy, and BET surface area analyzer. The testing results revealed that a slurry composition with 40 wt.% lithium aluminate was the optimal formulation to produce a good matrix with a pore size distribution of 0.1–0.4 μm and porosity of 50 vol.%. Coarse and fine LiAlO2 particles were added in the matrix slurry to enhance the mechanical strength. Several green sheets were heated and pressed to enhance the bulk density to get a dense matrix of MCFC. Finally, a single MCFC was assembled and tested. The testing results showed the matrix with 40% solid loading gave the maximum discharge current of 20 A at 0.56 V.

  7. Dye-sensitized solar cell and photocatalytic performance of nanocomposite photocatalyst prepared by electrochemical anodization

    Indian Academy of Sciences (India)

    MOHAMAD MOHSEN MOMENI

    2016-10-01

    This study compares different Fe-doped TiO$_2$ nanostructures in terms of their photocatalytic performance. Iron-doped TiO$_2$ nanostructures (FeTNs) were prepared by in situ anodizing of titanium in a single-stepprocess in the presence of 3, 9, 15 and 21 mM K$_3$Fe(CN)$_6$. Potassium ferricyanide was used as the iron source. Prepared films are amorphous, so these layers were thermally annealed. The effect of iron doping on the photoelectrochemical properties (including dye-sensitized solar cells) and photocatalysis properties (decomposition of methomyl) was investigated. In all investigated cases, the sample C, which was formed by anodizing in a ethylene glycol electrolyte containing 9 mM K$_3$Fe(CN)$_6$, exhibited better performance than the bare TiO$_2$ and FeNTs fabricated using other iron concentrations. This study demonstrated a feasible and simple anodizing method to fabricate an effective, reproducible and inexpensive photocatalyst for various applications.

  8. Film properties of alumina passivation layer for silicon solar cells prepared by spin-coating method

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Ryosuke, E-mail: rwatanabe@st.seikei.ac.jp; Kawashima, Mizuho; Saito, Yoji

    2015-09-01

    We prepared alumina passivation films deposited by a sol-gel wet process for silicon substrates. Aluminum acetylacetonate was used as a precursor, and the solution was spin-coated onto silicon substrates. Calcination temperature dependence of the passivation quality of the films was evaluated mainly by measuring effective lifetime using a photo conductance decay technique and capacitance–voltage measurements. Also, X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy were carried out to evaluate film properties. A large amount of negative fixed charge density (Q{sub f} = − 3.1 × 10{sup 12} cm{sup −2}) exists in the films calcined at 300 °C. On the other hand, a long effective lifetime of 400 μs was obtained for the sample calcined at 600 °C, and the passivation films had a large amount of positive fixed charge density (Q{sub f} = 3.6 × 10{sup 12} cm{sup −2}) with a low interface state density. - Highlights: • Alumina passivation films for silicon solar cells were prepared by spin-coating. • Electronic properties and the quality of passivation films were investigated. • Carrier lifetime was enhanced for the samples that were calcined above 400 °C. • The films calcined at 300 °C have high amount of negative fixed charge.

  9. Organic Photovoltaic Cells Based on PbPc Nanocolumns Prepared by Glancing Angle Deposition

    Directory of Open Access Journals (Sweden)

    Yang Liu

    2013-01-01

    Full Text Available Organic small material lead phthalocyanine (PbPc nanocolumns were prepared via glancing angle deposition (GLAD on indium tin oxide (ITO coated glass substrates. Organic electron acceptor materials fullerene (C60 was evaporated onto the nanocolumn PbPc thin films to prepare heterojunction structure ITO/PbPc/C60/Bphen/Al organic photovoltaic cells (OPVs. It is worthwhile to mention that C60 molecules firstly fill the voids between PbPc nanocolumns and then form impact C60 layer. The interpenetrating electron donor/acceptor structure effectively enhances interface between electron donor and electron acceptor, which is beneficial to exciton dissociation. The short circuit current density (Jsc of organic photovoltaic devices (OPVs based on PbPc nanocolumn was increased from 1.19 mA/cm2 to 1.74 mA/cm2, which should be attributed to the increase of interface between donor and acceptor. The effect of illumination intensity on the performance of OPVs was investigated by controlling the distance between light source and sample, and the Jsc of two kind of OPVs was increased along with the increase of illumination intensity.

  10. Silicon thin films prepared in the transition region and their use in solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, S.; Raniero, L.; Fortunato, E.; Aguas, H.; Ferreira, I.; Martins, R. [Department of Materials Science/CENIMAT, Faculty of Sciences and Technology, New University of Lisbon and CEMOP-UNINOVA, 2829-516 Caparica (Portugal); Liao, X.; Xu, Y.; Kong, G. [State Key Laboratory for Surface Physics, Institute of Semiconductors & amp; Center for Condensed Matter Physics, Chinese Academy of Sciences, Beijing 100083 (China)

    2006-11-23

    Diphasic silicon films (nc-Si/a-Si:H) have been prepared by a new regime of plasma enhanced chemical vapour deposition in the region adjacent of phase transition from amorphous to microcrystalline state. Comparing to the conventional amorphous silicon (a-Si:H), the nc-Si/a-Si:H has higher photoconductivity ({sigma}{sub ph}), better stability, and a broader light spectral response range in the longer wavelength range. It can be found from Raman spectra that there is a notable improvement in the medium range order. The blue shift for the stretching mode and red shift for the wagging mode in the IR spectra also show the variation of the microstructure. By using this kind of film as intrinsic layer, a p-i-n junction solar cell was prepared with the initial efficiency of 8.51% and a stabilized efficiency of 8.01% (AM1.5, 100mw/cm{sup 2}) at room temperature. (author)

  11. Improvement of MEAs for direct-methanol fuel cells by tuned layer preparation and coating technology

    Energy Technology Data Exchange (ETDEWEB)

    Lindermeir, A.; Rosenthal, G.; Kunz, U.; Hoffmann, U. [Institute for Chemical Process Engineering, Technical University of Clausthal, Leibnizstrasse 17, D-38678 Clausthal-Zellerfeld (Germany)

    2004-04-01

    Large scale production of membrane electrode assemblies (MEAs) for fuel cells requires a fabrication technology, which is not only based on electrochemical phenomena but also takes into account aspects of materials processing and conversion. For the fabrication of MEAs for DMFC a production sequence consisting of wet ball-milling, wet spray coating and calendering was developed at the ICVT Clausthal. This has a proven ability for the preparation of diffusion backings and catalyst layers with high reproducibility. To improve MEA performance, investigations concerning the influence of layer fabrication and composition are made with respect to the resulting morphology of the structures. Intermediates and MEAs are evaluated by conductivity measurements, determination of permeability values and current density-voltage characteristics supported by SEM and optical microscopy. (Abstract Copyright [2004], Wiley Periodicals, Inc.)

  12. Porous polylactic acid-silica hybrids: preparation, characterization, and study of mesenchymal stem cell osteogenic differentiation.

    Science.gov (United States)

    Pandis, Christos; Trujillo, Sara; Matos, Joana; Madeira, Sara; Ródenas-Rochina, Joaquín; Kripotou, Sotiria; Kyritsis, Apostolos; Mano, João F; Gómez Ribelles, José Luis

    2015-02-01

    A novel approach to reinforce polymer porous membranes is presented. In the prepared hybrid materials, the inorganic phase of silica is synthesized in-situ and inside the pores of aminolyzed polylactic acid (PLA) membranes by sol-gel reactions using tetraethylorthosilicate (TEOS) and glycidoxypropyltrimethoxysilane (GPTMS) as precursors. The hybrid materials present a porous structure with a silica layer covering the walls of the pores while GPTMS serves also as coupling agent between the organic and inorganic phase. The adjustment of silica precursors ratio allows the modulation of the thermomechanical properties. Culture of mesenchymal stem cells on these supports in osteogenic medium shows the expression of characteristic osteoblastic markers and the mineralization of the extracellular matrix.

  13. Successful Hematopoietic Stem Cell Transplantation Following a Cyclophosphamide-Containing Preparative Regimen with Concomitant Phenobarbital Administration

    Directory of Open Access Journals (Sweden)

    Catherine Weber

    2012-01-01

    Full Text Available Cyclophosphamide is an immunosuppressive agent and an anticancer prodrug which requires bioactivation catalyzed primarily by cytochrome P450 enzymes in order to be transformed into its active alkylating compounds. Concomitant administration of drugs known to inhibit or induce this enzyme system is a clinical concern. Herein, we present the case of a chronically ill 21-year-old patient who received high-dose cyclophosphamide, equine antithymocyte globulin (eATG, and total body irradiation (TBI followed by an allogeneic hematopoietic stem cell transplant (HSCT for severe aplastic anemia. Throughout her hospitalization, she continued to receive quadruple anticonvulsant therapy including phenobarbital for her long-standing seizure history. The preparative regimen was tolerated well aside from a hypersensitivity reaction to eATG, and minimal cyclophosphamide-related toxicities. Safe and effective administration of high-dose cyclophosphamide was possible with multidisciplinary care consisting of physician, nursing, pharmacy, neurology consultation, as well as social work and case management.

  14. Preparation of big size open-cell aluminum foam board using infiltration casting

    Institute of Scientific and Technical Information of China (English)

    Wang Lucai; Chen Yuyong; Wang Fang; Wu Jianguo; You Xiaohong

    2008-01-01

    This paper presents an infiltration casting technique for manufacturing big size open-cell aluminum foam boards. The principle and key technologies of infiltration casting are also analyzed. Based on the previous practice of the small size aluminum foam production, the die for preparing big size aluminum foam boards is designed and manufactured. The experiments on aluminum boards of 300 mm×300 mm×(20-75) mm, with the pore size ranging from 1.0 to 3.2 mm and average porosity of 60%, have been performed. The experimental results show that a reliable infiltration process depends critically on the pouring temperature of the molten AI-alloy, the preheated temperature of the mould and salt particles and vacuum. Current research explores the possibility of large-scale manufacturing and application of the aluminum foams.

  15. Preparation and properties of low-cost graphene counter electrodes for dye-sensitized solar cells

    Science.gov (United States)

    Wu, Qishuang; Shen, Yue; Wang, Qiandi; Gu, Feng; Cao, Meng; Wang, Linjun

    2013-12-01

    With the advantages of excellent electrical properties, high catalytic activity and low-cost preparation, Graphene is one of the most expected carbon materials to replace the expensive Pt as counter electrodes for dye-sensitized solar cells (DSSCs). In this paper, graphene counter electrodes were obtained by simple doctor-blade coating method on fluorine tin oxides (FTOs). The samples were investigated by X-ray diffraction (XRD), Raman spectroscopy and scanning electron microscope (SEM). Then the low-cost graphene electrodes were applied in typical sandwich-type DSSCs with TiO2 or ZnO as photoanodes, and their photoelectric conversion efficiency (η) were about 4.34% and 2.28%, respectively, which were a little lower than those of Pt electrodes but much higher than those of graphite electrodes. This law was consistent with the test results of electrochemical impedance spectroscopy (EIS). Low-cost graphene electrodes can be applied in DSSCs by process optimization.

  16. Preparation of Nanoporous TiO2 Electrodes for Dye-Sensitized Solar Cells

    OpenAIRE

    Hsiue-Hsyan Wang; Chaochin Su; Huei-Siou Chen; Yi-Cheng Liu; Yi-Wen Hsu; Nai-Mu Hsu; Wen-Ren Li

    2011-01-01

    Nano-porous TiO2 thin films have been widely used as the working electrodes in dye-sensitized solar cells (DSSCs). In this work, the phase-pure anatase TiO2 (a-TiO2) and rutile TiO2 (r-TiO2) have been prepared using hydrothermal processes. The investigation of photo-to-electron conversion efficiency of DSSCs fabricated from mixed-TiO2 with a-TiO2 and r-TiO2 ratio of 80 : 20 (A8R2) was performed and compared to that from commercial TiO2 (DP-25). The results showed higher efficiency of DSSC for...

  17. Appropriate materials and preparation techniques for polycrystalline-thin-film thermophotovoltaic cells

    Science.gov (United States)

    Dhere, Neelkanth G.

    1997-03-01

    Polycrystalline-thin-film thermophotovoltaic (TPV) cells have excellent potential for reducing the cost of TPV generators so as to address the hitherto inaccessible and highly competitive markets such as self-powered gas-fired residential warm air furnaces and energy-efficient electric cars, etc. Recent progress in polycrystalline-thin-film solar cells have made it possible to satisfy the diffusion length and intrinsic junction rectification criteria for TPV cells operating at high fluences. Continuous ranges of direct bandgaps of the ternary and pseudoternary compounds such as Hg1-xCdxTe, Pb1-xCdxTe, Hg1-xZnxTe, and Pb1-xZnxS cover the region of interest of 0.50-0.75 eV for efficient TPV conversion. Other ternary and pseudoternary compounds which show direct bandgaps in most of or all of the 0.50-0.75 eV range are Pb1-xZnxTe, Sn1-xCd2xTe2, Pb1-xCdxSe, Pb1-xZnxSe, and Pb1-xCdxS. Hg1-xCdxTe (with x~0.21) has been studied extensively for infrared detectors. PbTe and Pb1-xSnxTe have also been studied for infrared detectors. Not much work has been carried out on Hg1-xZnxTe thin films. Hg1-xCdxTe and Pb1-xCdxTe alloys cover a wide range of cut-off wavelengths from the far infrared to the near visible. Acceptors and donors are introduced in these materials by excess non-metal (Te) and excess metal (Hg and Pb) respectively. Extrinsic acceptor impurities are Cu, Au, and As while and In and Al are donor impurities. Hg1-xCdxTe thin films have been deposited by isothermal vapor-phase epitaxy (VPE), liquid phase epitaxy (LPE), hot-wall metalorganic chemical vapor deposition (MOCVD), electrodeposition, sputtering, molecular beam epitaxy (MBE), laser-assisted evaporation, and vacuum evaporation with or without hot-wall enclosure. The challenge in the preparation of Hg1-xCdxTe is to provide excess mercury incidence rate, to optimize the deposition parameters for enhanced mercury incorporation, and to achieve the requisite stoichiometry, grain size, and doping. MBE and MOCVD

  18. Comparative immune phenotypic analysis of cutaneous Squamous Cell Carcinoma and Intraepidermal Carcinoma in immune-competent individuals: proportional representation of CD8+ T-cells but not FoxP3+ Regulatory T-cells is associated with disease stage.

    Directory of Open Access Journals (Sweden)

    Andrew Freeman

    Full Text Available Squamous Cell Carcinoma (SCC is a type of non-melanoma skin cancer prevalent in immune-suppressed transplant recipients and older individuals with a history of chronic sun-exposure. SCC itself is believed to be a late-stage manifestation that can develop from premalignant lesions including Intraepidermal Carcinoma (IEC. Notably, while SCC regression is rare, IEC typically regresses in response to immune modifying topical treatments, however the underlying immunological reasons for these differential responses remain unclear. This study aimed to define whether IEC and SCC are associated with distinct immune profiles. We investigated the immune cell infiltrate of photo-damaged skin, IEC, and SCC tissue using 10-colour flow cytometry following fresh lesion digest. We found that IEC lesions contain higher percentages of CD3+ T-cells than photo-damaged skin, however, the abundance of CD3-CD56+ Natural Killer (NK cells, CD11c+HLA-DR+ conventional Dendritic Cells (cDC, BDCA-2+HLA-DR+ plasmacytoid DC (pDC, FoxP3+ Regulatory T-cells (T-reg, Vα24+Vβ11+ invariant NKT-cells, and γδ Tcells did not alter with disease stage. Within the total T-cell population, high percentages of CD4+ T-cells were associated with SCC, yet CD8+ T-cells were less abundant in SCC compared with IEC. Our study demonstrates that while IEC lesions contain a higher proportion of T-cells than SCC lesions in general, SCC lesions specifically display a lower abundance of CD8+ T-cells than IEC. We propose that differences in CD8+ T-cell abundance contribute critically to the different capacity of SCC and IEC to regress in response to immune modifying topical treatments. Our study also suggests that a high ratio of CD4+ T-cells to CD8+ T-cells may be a immunological diagnostic indicator of late-stage SCC development in immune-competent patients.

  19. Comparative immune phenotypic analysis of cutaneous Squamous Cell Carcinoma and Intraepidermal Carcinoma in immune-competent individuals: proportional representation of CD8+ T-cells but not FoxP3+ Regulatory T-cells is associated with disease stage.

    Science.gov (United States)

    Freeman, Andrew; Bridge, Jennifer A; Maruthayanar, Pirashanthini; Overgaard, Nana H; Jung, Ji-Won; Simpson, Fiona; Prow, Tarl W; Soyer, H Peter; Frazer, Ian H; Freeman, Michael; Wells, James W

    2014-01-01

    Squamous Cell Carcinoma (SCC) is a type of non-melanoma skin cancer prevalent in immune-suppressed transplant recipients and older individuals with a history of chronic sun-exposure. SCC itself is believed to be a late-stage manifestation that can develop from premalignant lesions including Intraepidermal Carcinoma (IEC). Notably, while SCC regression is rare, IEC typically regresses in response to immune modifying topical treatments, however the underlying immunological reasons for these differential responses remain unclear. This study aimed to define whether IEC and SCC are associated with distinct immune profiles. We investigated the immune cell infiltrate of photo-damaged skin, IEC, and SCC tissue using 10-colour flow cytometry following fresh lesion digest. We found that IEC lesions contain higher percentages of CD3+ T-cells than photo-damaged skin, however, the abundance of CD3-CD56+ Natural Killer (NK) cells, CD11c+HLA-DR+ conventional Dendritic Cells (cDC), BDCA-2+HLA-DR+ plasmacytoid DC (pDC), FoxP3+ Regulatory T-cells (T-reg), Vα24+Vβ11+ invariant NKT-cells, and γδ Tcells did not alter with disease stage. Within the total T-cell population, high percentages of CD4+ T-cells were associated with SCC, yet CD8+ T-cells were less abundant in SCC compared with IEC. Our study demonstrates that while IEC lesions contain a higher proportion of T-cells than SCC lesions in general, SCC lesions specifically display a lower abundance of CD8+ T-cells than IEC. We propose that differences in CD8+ T-cell abundance contribute critically to the different capacity of SCC and IEC to regress in response to immune modifying topical treatments. Our study also suggests that a high ratio of CD4+ T-cells to CD8+ T-cells may be a immunological diagnostic indicator of late-stage SCC development in immune-competent patients.

  20. Impact of individual intravenous iron preparations on the differentiation of monocytes towards macrophages and dendritic cells

    Science.gov (United States)

    Fell, Lisa H.; Seiler-Mußler, Sarah; Sellier, Alexander B.; Rotter, Björn; Winter, Peter; Sester, Martina; Fliser, Danilo; Heine, Gunnar H.; Zawada, Adam M.

    2016-01-01

    Background Treatment of iron deficiency with intravenous (i.v.) iron is a first-line strategy to improve anaemia of chronic kidney disease. Previous in vitro experiments demonstrated that different i.v. iron preparations inhibit differentiation of haematopoietic stem cells to monocytes, but their effect on monocyte differentiation to macrophages and mature dendritic cells (mDCs) has not been assessed. We investigated substance-specific effects of iron sucrose (IS), sodium ferric gluconate (SFG), ferric carboxymaltose (FCM) and iron isomaltoside 1000 (IIM) on monocytic differentiation to M1/M2 macrophages and mDCs. Methods Via flow cytometry and microRNA (miRNA) expression analysis, we morphologically and functionally characterized monocyte differentiation to M1/M2 macrophages and mDCs after monocyte stimulation with IS, SFG, FCM and IIM (0.133, 0.266 and 0.533 mg/mL, respectively). To assess potential clinical implications, we compared monocytic phagocytosis capacity in dialysis patients who received either 500 mg IS or IIM. Results Phenotypically, IS and SFG dysregulated the expression of macrophage (e.g. CD40, CD163) and mDC (e.g. CD1c, CD141) surface markers. Functionally, IS and SFG impaired macrophage phagocytosis capacity. Phenotypic and functional alterations were less pronounced with FCM, and virtually absent with IIM. In miRNA expression analysis of mDCs, IS dysregulated miRNAs such as miR-146b-5p and miR-155-5p, which are linked to Toll-like receptor and mitogen-activated protein kinase signalling pathways. In vivo, IS reduced monocytic phagocytosis capacity within 1 h after infusion, while IIM did not. Conclusions This study demonstrates that less stable i.v. iron preparations specifically affect monocyte differentiation towards macrophages and mDCs. PMID:27190361

  1. Dye sensitised solar cells with nickel oxide photocathodes prepared via scalable microwave sintering.

    Science.gov (United States)

    Gibson, Elizabeth A; Awais, Muhammad; Dini, Danilo; Dowling, Denis P; Pryce, Mary T; Vos, Johannes G; Boschloo, Gerrit; Hagfeldt, Anders

    2013-02-21

    Photoactive NiO electrodes for cathodic dye-sensitised solar cells (p-DSCs) have been prepared with thicknesses ranging between 0.4 and 3.0 μm by spray-depositing pre-formed NiO nanoparticles on fluorine-doped tin oxide (FTO) coated glass substrates. The larger thicknesses were obtained in sequential sintering steps using a conventional furnace (CS) and a newly developed rapid discharge sintering (RDS) method. The latter procedure is employed for the first time for the preparation of p-DSCs. In particular, RDS represents a scalable procedure that is based on microwave-assisted plasma formation that allows the production in series of mesoporous NiO electrodes with large surface areas for p-type cell photocathodes. RDS possesses the unique feature of transmitting heat from the bulk of the system towards its outer interfaces with controlled confinement of the heating zone. The use of RDS results in a drastic reduction of processing times with respect to other deposition methods that involve heating/calcination steps with associated reduced costs in terms of energy. P1-dye sensitized NiO electrodes obtained via the RDS procedure have been tested in DSC devices and their performances have been analysed and compared with those of cathodic DSCs derived from CS-deposited samples. The largest conversion efficiencies (0.12%) and incident photon-to-current conversion efficiencies, IPCEs (50%), were obtained with sintered NiO electrodes having thicknesses of ~1.5-2.0 μm. In all the devices, the photogenerated holes in NiO live significantly longer (τ(h) ~ 1 s) than have previously been reported for P1-sensitized NiO photocathodes. In addition, P1-sensitised sintered electrodes give rise to relatively high photovoltages (up to 135 mV) when the triiodide-iodide redox couple is used.

  2. A putatively phase variable gene (dca) required for natural competence in Neisseria gonorrhoeae but not Neisseria meningitidis is located within the division cell wall (dcw) gene cluster.

    Science.gov (United States)

    Snyder, L A; Saunders, N J; Shafer, W M

    2001-02-01

    A cluster of 18 open reading frames (ORFs), 15 of which are homologous to genes involved in division and cell wall synthesis, has been identified in Neisseria gonorrhoeae and Neisseria meningitidis. The three additional ORFs, internal to the dcw cluster, are not homologous to dcw-related genes present in other bacterial species. Analysis of the N. meningitidis strain MC58 genome for foreign DNA suggests that these additional ORFs have not been acquired by recent horizontal exchange, indicating that they are a long-standing, integral part of the neisserial dcw gene cluster. Reverse transcription-PCR analysis of RNA extracted from N. gonorrhoeae strain FA19 confirmed that all three ORFs are transcribed in gonococci. One of these ORFs (dca, for division cluster competence associated), located between murE and murF, was studied in detail and found to be essential for competence in the gonococcal but not in the meningococcal strains tested. Computer analysis predicts that dca encodes an inner membrane protein similar to hypothetical proteins produced by other gram-negative bacteria. In some meningococcal strains dca is prematurely terminated following a homopolymeric tract of G's, the length of which differs between isolates of N. meningitidis, suggesting that dca is phase variable in this species. A deletion and insertional mutation was made in the dca gene of N. gonorrhoeae strain FA19 and N. meningitidis strain NMB. This mutation abrogated the ability of the gonococci to be transformed with chromosomal DNA. Thus, we conclude that the dca-encoded gene product is an essential competence factor for gonococci.

  3. Pulpal regeneration after cavity preparation, with special reference to close spatio-relationships between odontoblasts and immunocompetent cells.

    Science.gov (United States)

    Ohshima, Hayato; Nakakura-Ohshima, Kuniko; Takeuchi, Kiichi; Hoshino, Masaaki; Takano, Yoshiro; Maeda, Takeyasu

    2003-04-01

    The regeneration process of the odontoblast cell layer incident to tooth injury, especially its relationship with immunocompetent cells in pulp healing, has not been fully understood. The purpose of the present study was to clarify this relationship between odontoblasts and immunocompetent cells in the process of pulp regeneration following cavity preparation in rat molars by immunocytochemistry for heat shock protein (Hsp) 25 as well as class II major histocompatibility complex (MHC) molecules. In untreated control teeth, intense Hsp 25-immunoreactivity was found in the cell bodies of odontoblasts and their processes within the predentin, whereas class II MHC-positive cells were predominantly located beneath the odontoblast cell layer. Cavity preparation caused the destruction of the odontoblast layer to form an edematous lesion and the shift of class II MHC-positive cells with the injured odontoblasts toward the pulp core at the affected site. Some damaged odontoblasts without apparent cytoplasmic processes, round in profile, retained the immunoreactivity for Hsp25, suggesting the survival of a part of the odontoblasts against artificial external stimuli. Twelve hours after cavity preparation, numerous class II MHC-positive cells appeared along the pulp-dentin border and extended their processes deep into the exposed dentinal tubules. By postoperative 72 hours, newly differentiated odontoblasts with Hsp 25-immunoreactivity were arranged at the pulp-dentin border, but the class II MHC-positive cells moved from the pulp-dentin border to the subodontoblastic layer. These findings indicate that the time course of changes in the expression of Hsp 25-immunoreactivity reflects the regeneration process of odontoblasts. The functional roles of Hsp 25-positive odontoblasts and immunocompetent cells such as class II MHC-positive cells in the process of pulp regeneration after cavity preparation are discussed in conjunction with our previous experimental data.

  4. Ultra-Fast and Optimized Method for the Preparation of Rodent Testicular Cells for Flow Cytometric Analysis

    Directory of Open Access Journals (Sweden)

    López-Carro Beatriz

    2009-01-01

    Full Text Available Abstract Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular tissue, various methods have been used to obtain enriched or purified cell populations, including flow cell sorting. Current protocols are usually time-consuming and may imply loss of short-lived RNAs, which is undesirable for expression profiling. We describe an optimized method to speed up the preparation of suitable testicular cell suspensions for cytometric analysis of different spermatogenic stages from rodents. The procedure takes only 15 min including testis dissection, tissue cutting, and processing through the Medimachine System (Becton Dickinson. This method could be a substitute for the more tedious and time-consuming cell preparation techniques currently in use.

  5. Ultra-Fast and Optimized Method for the Preparation of Rodent Testicular Cells for Flow Cytometric Analysis

    Directory of Open Access Journals (Sweden)

    Rodríguez-Casuriaga Rosana

    2009-03-01

    Full Text Available Abstract Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular tissue, various methods have been used to obtain enriched or purified cell populations, including flow cell sorting. Current protocols are usually time-consuming and may imply loss of short-lived RNAs, which is undesirable for expression profiling. We describe an optimized method to speed up the preparation of suitable testicular cell suspensions for cytometric analysis of different spermatogenic stages from rodents. The procedure takes only 15 min including testis dissection, tissue cutting, and processing through the Medimachine System (Becton Dickinson. This method could be a substitute for the more tedious and time-consuming cell preparation techniques currently in use.

  6. Human amniotic fluid derived cells can competently substitute dermal fibroblasts in a tissue-engineered dermo-epidermal skin analog

    NARCIS (Netherlands)

    Hartmann-Fritsch, Fabienne; Hosper, Nynke; Luginbuehl, Joachim; Biedermann, Thomas; Reichmann, Ernst; Meuli, Martin

    2013-01-01

    Human amniotic fluid comprises cells with high differentiation capacity, thus representing a potential cell source for skin tissue engineering. In this experimental study, we investigated the ability of human amniotic fluid derived cells to substitute dermal fibroblasts and support epidermis formati

  7. An Immune-Competent Murine Model to Study Elimination of AAV-Transduced Hepatocytes by Capsid-Specific CD8+ T Cells

    Directory of Open Access Journals (Sweden)

    Brett Palaschak

    2017-06-01

    Full Text Available Multiple independent adeno-associated virus (AAV gene therapy clinical trials for hemophilia B, utilizing different AAV serotypes, have reported a vector dose-dependent loss of circulating factor IX (FIX protein associated with capsid-specific CD8+ T cell (Cap-CD8 elimination of transduced hepatocytes. Hemophilia B patients who develop transient transaminitis and loss of FIX protein may be stabilized with the immune-suppressive (IS drug prednisolone, but do not all recover lost FIX expression, whereas some patients fail to respond to IS. We developed the first animal model demonstrating Cap-CD8 infiltration and elimination of AAV-transduced hepatocytes of immune-deficient mice. Here, we extend this model to an immune-competent host where Cap-CD8 transfer to AAV2-F9-treated mice significantly reduced circulating and hepatocyte FIX expression. Further, we studied two high-expressing liver tropic AAV2 variants, AAV2-LiA and AAV2-LiC, obtained from a rationally designed capsid library. Unlike AAV2, Cap-CD8 did not initially reduce circulating FIX levels for either variant. However, FIX levels were significantly reduced in AAV2-LiC-F9-treated, but not AAV2-LiA-F9-treated, mice at the study endpoint. Going forward, the immune-competent model may provide an opportunity to induce immunological memory directed against a surrogate AAV capsid antigen and study recall responses following AAV gene transfer.

  8. Preparation and Amplification of Colony of Goat Transgenic Fetal Fibroblast and Mammary Gland Epithelial Cell with Human Lactoferrin Gene

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yu-ling; LIU Feng-jun; ZHANG Yong

    2009-01-01

    [Objective] The aim was to explore technical system of making single transgenic positive cells become colony cells by amplification culture. [Method] Fetal fibroblasts and mammary gland epithelial cells of single goat fetus of pBLM-C1 which specifically expressed human lactoferrin were cloned. Single cell colony of single transfection cell was prepared with 3 concentrations of 0%, 50% and 100% conditioned culture media. Transfection cell and non-transfection cell were carried out amplification culture by con-culture, neo gene was as screened gene, genome DNA of transfection cell was detected by PCR method. Chromosome karyotype analysis of single colony cell was tested. [Result] Compared with non-conditioned culture medium, 100% conditioned culture medium could greatly increase survived rate of single colony cells (FF: 53.33% vs. 10.00%; MGE: 33.33% vs. 6.67%). Compared with control, con-culture of transfection cell and non-transfection cell could greatly increase rate of transfection cell single colony after amplification culture (FF: 53.33% vs. 10.00%;MGE: 33.33% vs. 6.67%), confluence time of amplification culture was significantly decreased (20-30 d). The result of PCR showed that the colony cell obtained by above method contained hLF target gene. The result of karyotype analysis showed that most cloned cell chromosomes were normal. [Conclusion] The study provides a reliable method for separating transgenic cell, inserting and diagnosing ideal vector, and can save expense and time for transgenic animal production.

  9. Application of stem-cell media to explant culture of human periosteum: An optimal approach for preparing osteogenic cell material

    Directory of Open Access Journals (Sweden)

    Kohya Uematsu

    2013-10-01

    Full Text Available As part of our clinical tests on bone regeneration using cultured periosteal sheets, here, we prepared cultured periosteal sheets in two types of stem-cell culture media, STK1 and STK3. Human periosteum was expanded either in 1% human serum–supplemented STK1 for 28 days, in 1% human serum–supplemented STK1 for 14 days followed by 1% human serum–supplemented STK3 for 14 days (1% human serum–supplemented STK1+3, or in 10% fetal bovine serum–supplemented Medium 199 for 28 days (control. Cultured periosteal sheet diameter and DNA content were significantly higher, and the multilayer structure was prominent in 1% human serum–supplemented STK1 and 1% human serum–supplemented STK1+3. The messenger RNA of osteoblastic markers was significantly upregulated in 1% human serum–supplemented STK1+3. Osteopontin-immunopositive staining and mineralization were evident across a wide area of the cultured periosteal sheet in 1% human serum–supplemented STK1+3. Subcutaneous implantation in nude mice following expansion in 1% human serum–supplemented STK1+3 produced the highest cultured periosteal sheet osteogenic activity. Expansion in 1% human serum–supplemented STK1+3 successfully induced cultured periosteal sheet growth while retaining osteogenic potential, and subsequent osteoblastic induction promoted the production of homogeneous cell material.

  10. Preparation of electron buffer layer with crystalline ZnO nanoparticles in inverted organic photovoltaic cells

    Science.gov (United States)

    Lee, Donghwan; Kang, Taeho; Choi, Yoon-Young; Oh, Seong-Geun

    2017-06-01

    Zinc oxide (ZnO) nanoparticles synthesized through sol-gel method were used to fabricate the electron buffer layer in inverted organic photovoltaic cells (OPVs) after thermal treatment. To investigate the effect of thermal treatment on the formation of crystalline ZnO nanoparticles, the amorphous ZnO nanoparticles were treated via hydrothermal method. The crystalline phase of ZnO with well-ordered structure could be obtained when the amorphous phase of ZnO was processed under hydrothermal treatment at 170 °C. The crystalline structure of ZnO thin film in inverted organic solar cell could be obtained under relatively low annealing temperature by using thermally treated ZnO nanoparticles. The OPVs fabricated by using crystalline ZnO nanoparticles for electron buffer layer exhibited higher efficiency than the conventional ZnO nanoparticles. The best power conversion efficiency (PCE) was achieved for 7.16% through the ZnO film using the crystalline ZnO nanoparticles. The proposed method to prepared ZnO nanoparticles (NPs) could effectively reduce energy consumption during the fabrication of OPVs, which would greatly contribute to advantages such as lower manufacturing costs, higher productivity and application on flexible substrates.

  11. Isoniazid Proliposome Powders for Inhalation—Preparation, Characterization and Cell Culture Studies

    Directory of Open Access Journals (Sweden)

    Wipaporn Rojanarat

    2011-07-01

    Full Text Available The aims of this study were to develop proliposome powders containing isoniazid (INH in a dry powder aerosol form. INH-proliposome powders were prepared by a spray drying method. Proliposome physicochemical properties were determined using cascade impactor, X-ray diffraction and differential scanning calorimetry. The toxicity of proliposomes to respiratory-associated cell lines and its potential to provoke immunological responses from alveolar macrophages (AM were determined. Free INH and INH-proliposome bioactivities were tested in vitro and in AM infected with Mycobacterium bovis (M. bovis. Aerosolization properties of INH-proliposome powders at 60 L/min, the powders showed mass median aerodynamic diameters of 2.99–4.92 mm, with fine particle fractions (aerosolized particles less than 4.4 µm of 15–35%. Encapsulation of INH was 18–30%. Proliposome formulations containing INH to mannitol ratios of 4:6 and 6:4 exhibited the greatest overlapping peak between the drug and mannitol. INH-proliposomes were evidently nontoxic to respiratory-associated cells, and did not activate AM to produce inflammatory mediators—including interleukin-1b (IL-1b, tumor necrosis factor-a (TNF-a, and nitric oxide—at a toxic level. The efficacy of INH-proliposome against AM infected with M. bovis was significantly higher than that of free INH (p < 0.05. INH-proliposomes are potential candidates for an alternative tuberculosis treatment.

  12. Isoniazid proliposome powders for inhalation-preparation, characterization and cell culture studies.

    Science.gov (United States)

    Rojanarat, Wipaporn; Changsan, Narumon; Tawithong, Ekawat; Pinsuwan, Sirirat; Chan, Hak-Kim; Srichana, Teerapol

    2011-01-01

    The aims of this study were to develop proliposome powders containing isoniazid (INH) in a dry powder aerosol form. INH-proliposome powders were prepared by a spray drying method. Proliposome physicochemical properties were determined using cascade impactor, X-ray diffraction and differential scanning calorimetry. The toxicity of proliposomes to respiratory-associated cell lines and its potential to provoke immunological responses from alveolar macrophages (AM) were determined. Free INH and INH-proliposome bioactivities were tested in vitro and in AM infected with Mycobacterium bovis (M. bovis). Aerosolization properties of INH-proliposome powders at 60 L/min, the powders showed mass median aerodynamic diameters of 2.99-4.92 μm, with fine particle fractions (aerosolized particles less than 4.4 μm) of 15-35%. Encapsulation of INH was 18-30%. Proliposome formulations containing INH to mannitol ratios of 4:6 and 6:4 exhibited the greatest overlapping peak between the drug and mannitol. INH-proliposomes were evidently nontoxic to respiratory-associated cells, and did not activate AM to produce inflammatory mediators-including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide-at a toxic level. The efficacy of INH-proliposome against AM infected with M. bovis was significantly higher than that of free INH (p < 0.05). INH-proliposomes are potential candidates for an alternative tuberculosis treatment.

  13. Preparation of a self-humidifying membrane electrode assembly for fuel cell and its performance analysis

    Institute of Scientific and Technical Information of China (English)

    王诚; 毛宗强; 徐景明; 谢晓峰; 杨立寨

    2003-01-01

    A novel nano-porous material SiO2-gel was prepared. After being purified by H2O2, then protonized by H2SO4 and desiccated in vacuum, the SiO2-gel, mixed with Nafion solution, was coated between an electrode and a solid electrolyte, which made a new type of self-humidifying membrane electrode assembly. The SiO2 powder was characterized by FTIR, BET and XRD. The surface of the electrodes was characterized by SEM and EDS. The performances of the self-hu- midifying membrane electrodes were analyzed by polarization discharge and AC impedance under the operation modes of external humidification and self-humidification respectively. Experimental results indicated that the SiO2 powder held super-hydrophilicity, and the layer of SiO2 and Nafion polymer between electrode and solid electrolyte expanded three-dimension electrochemistry reaction area, maintained stability of catalyst layer and enhanced back-diffusion of water from cathode to anode, so the PEM Fuel cell can generate electricity at self-humidification mode. The power density of single PEM fuel cell reached 1.5 W/cm2 under 0.2 Mpa, 70℃ and dry hydrogen and oxygen.

  14. PREPARATION AND CHARACTERIZATION OF PVA BASED SOLID POLYMER ELECTROLYTES FOR ELECTROCHEMICAL CELL APPLICATIONS

    Institute of Scientific and Technical Information of China (English)

    Anji Reddy Polu; Ranveer Kumar

    2013-01-01

    Solid polymer electrolyte films containing poly(vinyl alcohol) (PVA) and magnesium nitrate (Mg(NO3)2) were prepared by solution casting technique and characterized by using XRD,FT1R,DSC and AC impedance spectroscopic analysis.The amorphous nature of the polymer electrolyte films has been confirmed by XRD.The complex formation between PVA and Mg salt has been confirmed by FTIR.The glass transition temperature decreases with increasing the Mg salt concentration.The AC impedance studies are performed to evaluate the ionic conductivity of the polymer electrolyte films in the range of 303-383 K,and the temperature dependence seems to obey the Arrhenius behavior.Transport number measurements show that the charge transport is mainly due to ions.Electrochemical cell of configuration Mg/(PVA + Mg(NO3)2) (70:30)/(I2 + C + electrolyte) has been fabricated.The discharge characteristics of the cell were studied for a constant load of 100 kΩ.

  15. Optimized extract preparation methods and reaction conditions for improved yeast cell-free protein synthesis.

    Science.gov (United States)

    Hodgman, C Eric; Jewett, Michael C

    2013-10-01

    Cell-free protein synthesis (CFPS) has emerged as a powerful platform technology to help satisfy the growing demand for simple, affordable, and efficient protein production. In this article, we describe a novel CFPS platform derived from the popular bio-manufacturing organism Saccharomyces cerevisiae. By developing a streamlined crude extract preparation protocol and optimizing the CFPS reaction conditions we were able to achieve active firefly luciferase synthesis yields of 7.7 ± 0.5 µg mL(-1) with batch reactions lasting up to 2 h. This duration of synthesis is the longest ever reported for a yeast CFPS batch reaction. Furthermore, by removing extraneous processing steps and eliminating expensive reagents from the cell-free reaction, we have increased relative product yield (µg protein synthesized per $ reagent cost) over an alternative commonly used method up to 2000-fold from ∼2 × 10(-4) to ∼4 × 10(-1)  µg $(-1) , which now puts the yeast CPFS platform on par with other eukaryotic CFPS platforms commercially available. Our results set the stage for developing a yeast CFPS platform that provides for high-yielding and cost-effective expression of a variety of protein therapeutics and protein libraries.

  16. Application of green chemistry techniques to prepare electrocatalysts for direct methanol fuel cells.

    Science.gov (United States)

    Shimizu, Kenichi; Wang, Joanna S; Wai, Chien M

    2010-03-25

    A series of green techniques for synthesizing carbon nanotube-supported platinum nanoparticles and their high electrocatalytic activity toward methanol fuel cell applications are reported. The techniques utilize either the supercritical fluid carbon dioxide or water as a medium for depositing platinum nanoparticles on surfaces of multiwalled or single-walled carbon nanotubes. The catalytic properties of the carbon nanotubes-supported Pt nanoparticle catalysts prepared by four different techniques are compared for anodic oxidation of methanol and cathodic reduction of oxygen using cyclic voltammetry. One technique using galvanic exchange of Pt(2+) in water with zerovalent iron present on the surfaces of as-grown single-walled carbon nanotubes produces a Pt catalyst that shows an unusually high catalytic activity for reduction of oxygen but a negligible activity for oxidation of methanol. This fuel-selective catalyst may have a unique application as a cathode catalyst in methanol fuel cells to alleviate the problems caused by crossover of methanol through the polymer electrolyte membrane.

  17. Preparation of tumor antigen-loaded mature dendritic cells for immunotherapy.

    Science.gov (United States)

    Sabado, Rachel Lubong; Miller, Elizabeth; Spadaccia, Meredith; Vengco, Isabelita; Hasan, Farah; Bhardwaj, Nina

    2013-08-01

    While clinical studies have established that antigen-loaded DC vaccines are safe and promising therapy for tumors, their clinical efficacy remains to be established. The method described below, prepared in accordance with Good Manufacturing Process (GMP) guidelines, is an optimization of the most common ex vivo preparation method for generating large numbers of DCs for clinical studies. Our method utilizes the synthetic TLR 3 agonist Polyinosinic-Polycytidylic Acid-poly-L-lysine Carboxymethylcellulose (Poly-ICLC) to stimulate the DCs. Our previous study established that Poly-ICLC is the most potent individual maturation stimulus for human DCs as assessed by an upregulation of CD83 and CD86, induction of interleukin-12 (IL-12), tumor necrosis factor (TNF), interferon gamma-induced protein 10 (IP-10), interleukmin 1 (IL-1), and type I interferons (IFN), and minimal interleukin 10 (IL-10) production. DCs are differentiated from frozen peripheral blood mononuclear cells (PBMCs) obtained by leukapheresis. PBMCs are isolated by Ficoll gradient centrifugation and frozen in aliquots. On Day 1, PBMCs are thawed and plated onto tissue culture flasks to select for monocytes which adhere to the plastic surface after 1-2 hr incubation at 37 °C in the tissue culture incubator. After incubation, the lymphocytes are washed off and the adherent monocytes are cultured for 5 days in the presence of interleukin-4 (IL-4) and granulocyte macrophage-colony stimulating factor (GM-CSF) to differentiate to immature DCs. On Day 6, immature DCs are pulsed with the keyhole limpet hemocyanin (KLH) protein which serves as a control for the quality of the vaccine and may boost the immunogenicity of the vaccine. The DCs are stimulated to mature, loaded with peptide antigens, and incubated overnight. On Day 7, the cells are washed, and frozen in 1 ml aliquots containing 4-20 x 10(6) cells using a controlled-rate freezer. Lot release testing for the batches of DCs is performed and must meet minimum

  18. Evaluation of the Efficiency of Different Disruption Methods on Yeast Cell Wall Preparation for β-Glucan Isolation

    Directory of Open Access Journals (Sweden)

    Anna Bzducha-Wróbel

    2014-12-01

    Full Text Available Selected methods for yeast cell disruption were evaluated to establish their suitability for cell wall preparation in the process of β-glucan isolation. The effect of different disruption methods on contents of total saccharides, β-glucans and proteins in the produced cell walls preparations was analyzed. The degree of cell wall purification from intracellular components was established on the basis of the ratio of solubilised material. The investigated methods included: cell exposure to hot water (autoclaving, thermally-induced autolysis, homogenization in a bead mill, sonication and their combinations. Experimental systems were prepared in water (pH 5.0 and pH 7.0 and Tris-HCl buffer (pH 8.0. The Saccharomyces cerevisiae yeast cell wall preparations with the highest degree of cytosol component release and purification of β-glucans were produced by 30 min of cell homogenization with zirconium-glass beads (0.5 mm in diameter. This was confirmed by the highest ratio of solubilised material (approx. 64%–67%. The thus-produced preparations contained ca. 60% of total saccharides, 13%–14% of β(1,3/(1,6-glucans, and approx. 35% of crude proteins. Similar results were obtained after autolysis coupled with bead milling as well as with sonication, but the time required for these processes was more than 24 h. Homogenization in a bead mill could be valuable for general isolation procedures because allows one to eliminate the different autolytic activity of various yeast strains.

  19. Cryopreservation in Closed Bag Systems as an Alternative to Clean Rooms for Preparations of Peripheral Blood Stem Cells.

    Science.gov (United States)

    Spoerl, Silvia; Peter, Robert; Krackhardt, Angela M

    2016-01-01

    Autologous and allogeneic stem cell transplantation (SCT) represents a therapeutic option widely used for hematopoietic malignancies. One important milestone in the development of this treatment strategy was the development of effective cryopreservation technologies resulting in a high quality with respect to cell viability as well as lack of contamination of the graft.Stem cell preparations have been initially performed within standard laboratories as it is routinely still the case in many countries. With the emergence of cleanrooms, manufacturing of stem cell preparations within these facilities has become a new standard mandatory in Europe. However, due to high costs and laborious procedures, novel developments recently emerged using closed bag systems as reliable alternatives to conventional cleanrooms. Several hurdles needed to be overcome including the addition of the cryoprotectant dimethylsulfoxide (DMSO) as a relevant manipulation. As a result of the development, closed bag systems proved to be comparable in terms of product quality and patient outcome to cleanroom products. They also comply with the strict regulations of good manufacturing practice.With closed systems being available, costs and efforts of a cleanroom facility may be substantially reduced in the future. The process can be easily extended for other cell preparations requiring minor modifications as donor lymphocyte preparations. Moreover, novel developments may provide solutions for the production of advanced-therapy medicinal products in closed systems.

  20. Preparation, Characterization and Tests of Incorporation in Stem Cells of Superparamagnetic Iron Oxide

    Science.gov (United States)

    Haddad, P. S.; Britos, T. N.; Li, L. M.; Li, L. D. S.

    2015-05-01

    Superparamagnetic iron oxide nanoparticles (SPIONs) have been produced and used as contrast-enhancing agents in magnetic resonance imaging (MRI) for diagnostic use in a wide range of maladies including cardiovascular, neurological disorders, and cancer. The reasons why these SPIONs are attractive for medical purposes are based on their important and unique features. The large surface area of the nanoparticles and their manipulation through an external magnetic field are features that allow their use for carrying a large number of molecules such as biomolecules or drugs. In this scenario, the present work reports on the synthesis and characterization of SPIONs and in vitro MRI experiments to increase their capacity as probes for MRI applications on stem cells therapy. Initially, the SPIONs were prepared through the co-precipitation method using ferrous and ferric chlorides in acidic solution. The SPIONs were coated with two thiolmolecules such as mercaptosuccinic acid (MSA) and cysteine (Cys) (molar ratio SPIONs:ligand = 1:20), leading to the formation of a stable aqueous dispersion of thiolated nanoparticles (SH-SPIONs). The SH-SPIONs were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and vibrating sample magnetometry (VSM). The results showed that the SH-SPIONs have a mean diameter of 14 nm and display superparamagnetic behavior at room temperature. Preliminary tests of incorporation of SH-SPIONs were evaluated stem cells. The results showed that the thiolated nanoparticles have no toxic effects for stem cells and successfully internalized and enhance the contrast in MRI.

  1. One Health Core Competency Domains.

    Science.gov (United States)

    Frankson, Rebekah; Hueston, William; Christian, Kira; Olson, Debra; Lee, Mary; Valeri, Linda; Hyatt, Raymond; Annelli, Joseph; Rubin, Carol

    2016-01-01

    The emergence of complex global challenges at the convergence of human, animal, and environmental health has catalyzed a movement supporting "One Health" approaches. Despite recognition of the importance of One Health approaches to address these complex challenges, little effort has been directed at identifying the seminal knowledge, skills, and attitudes necessary for individuals to successfully contribute to One Health efforts. Between 2008 and 2011, three groups independently embarked on separate initiatives to identify core competencies for professionals involved with One Health approaches. Core competencies were considered critically important for guiding curriculum development and continuing professional education, as they describe the knowledge, skills, and attitudes required to be effective. A workshop was convened in 2012 to synthesize the various strands of work on One Health competencies. Despite having different mandates, participants, and approaches, all of these initiatives identified similar core competency domains: management; communication and informatics; values and ethics; leadership; teams and collaboration; roles and responsibilities; and systems thinking. These core competency domains have been used to develop new continuing professional education programs for One Health professionals and help university curricula prepare new graduates to be able to contribute more effectively to One Health approaches.

  2. One Health Core Competency Domains

    Science.gov (United States)

    Frankson, Rebekah; Hueston, William; Christian, Kira; Olson, Debra; Lee, Mary; Valeri, Linda; Hyatt, Raymond; Annelli, Joseph; Rubin, Carol

    2016-01-01

    The emergence of complex global challenges at the convergence of human, animal, and environmental health has catalyzed a movement supporting “One Health” approaches. Despite recognition of the importance of One Health approaches to address these complex challenges, little effort has been directed at identifying the seminal knowledge, skills, and attitudes necessary for individuals to successfully contribute to One Health efforts. Between 2008 and 2011, three groups independently embarked on separate initiatives to identify core competencies for professionals involved with One Health approaches. Core competencies were considered critically important for guiding curriculum development and continuing professional education, as they describe the knowledge, skills, and attitudes required to be effective. A workshop was convened in 2012 to synthesize the various strands of work on One Health competencies. Despite having different mandates, participants, and approaches, all of these initiatives identified similar core competency domains: management; communication and informatics; values and ethics; leadership; teams and collaboration; roles and responsibilities; and systems thinking. These core competency domains have been used to develop new continuing professional education programs for One Health professionals and help university curricula prepare new graduates to be able to contribute more effectively to One Health approaches. PMID:27679794

  3. One Health Core Competency Domains

    Directory of Open Access Journals (Sweden)

    Rebekah Frankson

    2016-09-01

    Full Text Available The emergence of complex global challenges at the convergence of human, animal, and environmental health has catalyzed a movement supporting ‘One Health’ approaches. Despite recognition of the importance of One Health approaches to address these complex challenges, little effort has been directed at identifying the seminal knowledge, skills and attitudes necessary for individuals to successfully contribute to One Health efforts. Between 2008 and 2011, three groups independently embarked on separate initiatives to identify core competencies for professionals involved with One Health approaches. Core competencies were considered critically important for guiding curriculum development and continuing professional education as they describe the knowledge, skills and attitudes required to be effective. A workshop was convened in 2012 to synthesize the various strands of work on One Health competencies. Despite having different mandates, participants, and approaches, all of these initiatives identified similar core competency domains: management; communication and informatics; values and ethics; leadership; teams and collaboration; roles and responsibilities; and systems thinking. These core competency domains have been used to develop new continuing professional education programs for One Health professionals and help university curricula prepare new graduates to be able to contribute more effectively to One Health approaches.

  4. Examining Rhodium Catalyst complexes for Use with Conducting Polymers Designed for Fuel Cells in Preparing Biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Carpio, M.M.; Kerr, J.B.

    2005-01-01

    Biosensing devices are important because they can detect, record, and transmit information regarding the presence of, or physiological changes in, different chemical or biological materials in the environment. The goal of this research is to prepare a biosensing device that is effective, quick, and low cost. This is done by examining which chemicals will work best when placed in a biosensor. The first study involved experimenting on a rhodium catalyst complexed with ligands such as bipyridine and imidazole. The rhodium catalyst is important because it is reduced from RhIII to RhI, forms a hydride by reaction with water and releases the hydride to react with nicotinamide adenine dinucleotide (NAD+) to selectively produce 1,4-NADH, the reduced form of NAD+. The second study looked at different types of ketones and enzymes for the enzyme-substrate reaction converting a ketone into an alcohol. Preliminary results showed that the rhodium complexed with bipyridine was able to carry out all the reactions, while the rhodium complexed with imidazole was not able to produce and release hydrides. In addition, the most effective ketone to use is benzylacetone with the enzyme alcohol dehydrogenase from baker’s yeast. Future work includes experimenting with bis-imidazole, which mimics the structure of bipyridine to see if it has the capability to reduce and if the reduction rate is comparable to the bipyridine complex. Once all testing is completed, the fastest catalysts will be combined with polymer membranes designed for fuel cells to prepare biosensing devices that can be used in a variety of applications including ones in the medical and environmental fields.

  5. Human amniotic fluid derived cells can competently substitute dermal fibroblasts in a tissue-engineered dermo-epidermal skin analog

    OpenAIRE

    2013-01-01

    PURPOSE: Human amniotic fluid comprises cells with high differentiation capacity, thus representing a potential cell source for skin tissue engineering. In this experimental study, we investigated the ability of human amniotic fluid derived cells to substitute dermal fibroblasts and support epidermis formation and stratification in a humanized animal model. METHODS: Dermo-epidermal skin grafts with either amniocytes or with fibroblasts in the dermis were compared in a rat model. Full-thicknes...

  6. Pin-Hole Free Perovskite Film for Solar Cells Application Prepared by Controlled Two-Step Spin-Coating Method

    Science.gov (United States)

    Bahtiar, A.; Rahmanita, S.; Inayatie, Y. D.

    2017-05-01

    Morphology of perovskite film is a key important for achieving high performance perovskite solar cells. Perovskite films are commonly prepared by two-step spin-coating method. However, pin-holes are frequently formed in perovskite films due to incomplete conversion of lead-iodide (PbI2) into perovskite CH3NH3PbI3. Pin-holes in perovskite film cause large hysteresis in current-voltage curve of solar cells due to large series resistance between perovskite layer-hole transport material. Moreover, crystal structure and grain size of perovskite crystal are also other important parameters for achieving high performance solar cells, which are significantly affected by preparation of perovskite film. We studied the effect of preparation of perovskite film using controlled spin-coating parameters on crystal structure and morphological properties of perovskite film. We used two-step spin-coating method for preparation of perovskite film with varied spinning speed, spinning time and temperature of spin-coating process to control growth of perovskite crystal aimed to produce high quality perovskite crystal with pin-hole free and large grain size. All experiment was performed in air with high humidity (larger than 80%). The best crystal structure, pin-hole free with large grain crystal size of perovskite film was obtained from film prepared at room temperature with spinning speed 1000 rpm for 20 seconds and annealed at 100°C for 300 seconds.

  7. Immune competence of cancer-reactive T cells generated de novo in adult tumor-bearing mice.

    Science.gov (United States)

    May, Kenneth F; Lute, Kenneth; Kocak, Ergun; Abdessalam, Shahab; Yin, Lijie; Li, Ou; Guan, Zhen; Philips, Gary; Zheng, Pan; Liu, Yang

    2007-01-01

    The impact of timing of antigen introduction into fetus and neonates leads to the suggestion that pre-existing antigens are tolerogenic to immunocompetent cells generated thereafter. This hypothesis predicts that in patients with cancer who are undergoing bone marrow transplantation, newly produced T cells with specificity for pre-existing tumor cells will be inactivated by the tumor antigens in the host. Because the effect of tumor cells on developing cancer-reactive T cells has not been investigated, we set out to systematically analyze the impact of tumor cells in the periphery on the development of tumor-reactive T cells in the thymus and their immunocompetence in the periphery. Our data demonstrate that in the host in which a tumor is established in the periphery, the cancer-reactive T cells develop normally, remain fully immunocompetent, become activated in the periphery, and cause regression of large established tumors. The immunocompetence of T cells generated in an antigen-bearing host is also confirmed in a skin graft transplantation model.

  8. Modification of aflatoxin B1 and ochratoxin A toxicokinetics in rats administered a yeast cell wall preparation

    OpenAIRE

    2010-01-01

    Abstract The cell wall of Saccharomyces cerevisiae can bind mycotoxins in vitro but there is scarce information on whether this property decreases the absorption of mycotoxins in vivo. The effect of a yeast cell wall preparation (YCW) on toxicokinetics and balance excretion (urine and faeces) of aflatoxin B1 (AFB1) and ochratoxin A (OTA) was tested in rats after oral administration of each toxin. The 3H-labelled mycotoxins were used at low doses. Co-administration of YCW with AF...

  9. Induction of apoptosis in MCF-7 cells by β-1,3-xylooligosaccharides prepared from Caulerpa lentillifera.

    Science.gov (United States)

    Maeda, Reiko; Ida, Tomoaki; Ihara, Hideshi; Sakamoto, Tatsuji

    2012-01-01

    β-1,3-Xylan was prepared from the green alga, Caulerpa lentillifera, and hydrolyzed to oligosaccharides by a mild acid treatment. The average degree of polymerization was about 5. The oligosaccharides reduced the number of viable human breast cancer MCF-7 cells in a dose-dependent manner, and induced chromatin condensation and degradation of poly ADP-ribose polymerase, indicating that they induced apoptosis in MCF-7 cells.

  10. Cultural competencies for graduate nursing education.

    Science.gov (United States)

    Clark, Lauren; Calvillo, Evelyn; Dela Cruz, Felicitas; Fongwa, Marie; Kools, Susan; Lowe, John; Mastel-Smith, Beth

    2011-01-01

    Nursing is challenged to meet the health needs of ethnic and socioculturally diverse populations. To this end, American Association of Colleges of Nursing (AACN) charged an expert nursing faculty advisory group to formulate competencies for graduate nursing education, expanding them to integrate leadership and scholarship. The Cultural Competency in Baccalaureate Nursing Education served as the springboard for the initiative. In formulating the graduate cultural competencies and the toolkit, the advisory group reviewed all AACN Essentials documents and the cultural competency literature, drew upon their collective experiences with cultural diversity, and used cultural humility as the supporting framework. Six core competencies were formulated and endorsed by the AACN board of directors and key professional nursing organizations. A companion toolkit was compiled to provide resources for the implementation of the competencies. A 1-day conference was held in California to launch the cultural competencies and toolkit. Dissemination to graduate nursing programs is in process, with emphasis on faculty readiness to undertake this graduate educational transformation. The AACN Cultural Competencies for Graduate Nursing Education set national standards to prepare culturally competent nurses at the graduate level who will contribute to the elimination of health disparities through education, clinical practice, research, scholarship, and policy.

  11. The effect of conspecific ampulla oviductal epithelial cells during in vitro maturation on oocyte developmental competence and maturation-promoting factor (MPF) activity in sheep.

    Science.gov (United States)

    Dadashpour Davachi, Navid; Kohram, Hamid; Zare Shahneh, Ahmad; Zhandi, Mahdi; Goudarzi, Abbas; Fallahi, Roozbeh; Masoudi, Reza; Yousefi, Ali Reza; Bartlewski, Pawel M

    2017-01-15

    The acquisition of fertilization ability by oocytes is one of the prerequisites for successful in vitro embryo production. In the present study, we examined the influence of conspecific ampulla oviductal epithelial cells incubated with cumulus-oocyte complexes (COCs) throughout the IVM phase on the developmental competence and maturation-promoting factor (MPF) activity of sheep oocytes. There were six experimental groups in this study, namely four groups with and two groups without oviductal epithelial cells added to IVM media: adult COCs matured in vitro with the ampulla oviductal epithelial cells obtained from adult (AAE; G1) or prepubertal donors (prepubertal sheep ampulla oviductal epithelial cells [PAE]; G4), COCs obtained from prepubertal animals cocultured with AAE (G2) or PAE (G3), and adult (C1) and prepubertal sheep COCs (C2) matured without oviductal epithelial cells. Coincubation of oocytes retrieved from both adult and sexually immature donors with AAE (G1 and G2) resulted in significantly improved rates of metaphase-II (M-II) attainment (G1: 85.1 ± 2.0 and G2: 40.2 ± 1.3) and blastocyst formation (G1: 42.2 ± 1.1 and G2: 21.2 ± 1.0) as well as blastocyst development (total cell count; G1: 130.3 ± 7.8, G2: 70.2 ± 3.5) compared with their respective controls (C1: 94.3 ± 4.1 and C2: 49.7 ± 2.0). Prior to IVM, the activity of MPF was greater (P vitro embryo production efficiency. A significant increase in MPF activity following IVM of G2 oocytes could be responsible, at least partly, for the improved rate of blastocyst formation after IVF of prepubertal sheep oocytes. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

  12. Comparison of dengue virus type 2-specific small RNAs from RNA interference-competent and -incompetent mosquito cells.

    Directory of Open Access Journals (Sweden)

    Jaclyn C Scott

    Full Text Available The exogenous RNA interference (RNAi pathway is an important antiviral defense against arboviruses in mosquitoes, and virus-specific small interfering (siRNAs are key components of this pathway. Understanding the biogenesis of siRNAs in mosquitoes could have important ramifications in using RNAi to control arbovirus transmission. Using deep sequencing technology, we characterized dengue virus type 2 (DENV2-specific small RNAs produced during infection of Aedes aegypti mosquitoes and A. aegypti Aag2 cell cultures and compared them to those produced in the C6/36 Aedes albopictus cell line. We show that the size and mixed polarity of virus-specific small RNAs from DENV-infected A. aegypti cells indicate that they are products of Dicer-2 (Dcr2 cleavage of long dsRNA, whereas C6/36 cells generate DENV2-specific small RNAs that are longer and predominantly positive polarity, suggesting that they originate from a different small RNA pathway. Examination of virus-specific small RNAs after infection of the two mosquito cell lines with the insect-only flavivirus cell fusing agent virus (CFAV corroborated these findings. An in vitro assay also showed that Aag2 A. aegypti cells are capable of siRNA production, while C6/36 A. albopictus cells exhibit inefficient Dcr2 cleavage of long dsRNA. Defective expression or function of Dcr2, the key initiator of the RNAi pathway, might explain the comparatively robust growth of arthropod-borne viruses in the C6/36 cell line, which has been used frequently as a surrogate for studying molecular interactions between arboviruses and cells of their mosquito hosts.

  13. Conversion of dechlorodauricumine into miharumine by a cell-free preparation from cultured roots of Menispermum dauricum.

    Science.gov (United States)

    Hori, Rieko; Sugimoto, Gen; Matsui, Miharu; Yamauchi, Yasuo; Takikawa, Hirosato; Sugimoto, Yukihiro

    2009-02-01

    Dechlorodauricumine (5) and dechloroacutumine (6) were converted to miharumine (7) and dechloroacutumidine (8), respectively, by a cell-free preparation from cultured roots of Menispermum dauricum in the presence of FAD. The structures of 7 and 8 were elucidated on the basis of spectroscopic analyses and chemical conversion.

  14. Influence of interferon preparations on the proliferative capacity of human and mouse bone marrow cells in vitro

    NARCIS (Netherlands)

    E. van 't Hull (Eveline); B. Löwenberg (Bob); M.J. de Vries (Marco); H. Schellekens (Huub)

    1978-01-01

    textabstractThe toxicity of interferon to bone marrow was studied by the use of in vitro colony forming assays for hemopoietic cells. In the same study the relative inhibitory effects of two clinically common interferon preparations, leukocyte and fibroblast interferons, were compared with regard to

  15. Preparation, characterisation and application of Pd/C nanocatalyst in passive alkaline direct ethanol fuel cells (ADEFC)

    CSIR Research Space (South Africa)

    Modibedi, RM

    2015-10-01

    Full Text Available This work describes the use of Pd nanocatalyst on Vulcan XC-72 (at a very low loading, 8 wt % Pd) in passive alkaline direct ethanol fuel cell (ADEFC). The average Pd particle size obtained with TEM was 3.5 ± 0.5 nm. The XRD results of the prepared...

  16. Influence of interferon preparations on the proliferative capacity of human and mouse bone marrow cells in vitro

    NARCIS (Netherlands)

    E. van 't Hull (Eveline); B. Löwenberg (Bob); M. de Vries (Marco); H. Schellekens (Huub)

    1978-01-01

    textabstractThe toxicity of interferon to bone marrow was studied by the use of in vitro colony forming assays for hemopoietic cells. In the same study the relative inhibitory effects of two clinically common interferon preparations, leukocyte and fibroblast interferons, were compared with regard to

  17. Combining designer cells and click chemistry for a one-pot four-step preparation of enantiopure beta-hydroxytriazoles

    NARCIS (Netherlands)

    Szymanski, Wiktor; Postema, Christiaan; Tarabiono, C.; Berthiol, Florian; Campbell-Verduyn, Lachlan; de Wildeman, S.; de Vries, J.G.; Feringa, B.L.; Janssen, D.B.

    2010-01-01

    The multistep catalytic process using designer cells, either added as freshly prepared suspensions or as stable lyophilized powder, and click reaction can be performed in one pot. The sequence of four reactions allows the production of both enantiomers of beta-hydroxytriazoles with high enantiomeric

  18. Combining designer cells and click chemistry for a one-pot four-step preparation of enantiopure beta-hydroxytriazoles

    NARCIS (Netherlands)

    Szymanski, Wiktor; Postema, Christiaan; Tarabiono, C.; Berthiol, Florian; Campbell-Verduyn, Lachlan; de Wildeman, S.; de Vries, J.G.; Feringa, B.L.; Janssen, D.B.

    2010-01-01

    The multistep catalytic process using designer cells, either added as freshly prepared suspensions or as stable lyophilized powder, and click reaction can be performed in one pot. The sequence of four reactions allows the production of both enantiomers of beta-hydroxytriazoles with high enantiomeric

  19. Design of a disulfide-less, pharmacologically inert, and chemically competent analog of maurocalcine for the efficient transport of impermeant compounds into cells.

    Science.gov (United States)

    Ram, Narendra; Weiss, Norbert; Texier-Nogues, Isabelle; Aroui, Sonia; Andreotti, Nicolas; Pirollet, Fabienne; Ronjat, Michel; Sabatier, Jean-Marc; Darbon, Hervé; Jacquemond, Vincent; De Waard, Michel

    2008-10-03

    Maurocalcine is a 33-mer peptide initially isolated from the venom of a Tunisian scorpion. It has proved itself valuable as a pharmacological activator of the ryanodine receptor and has helped the understanding of the molecular basis underlying excitation-contraction coupling in skeletal muscles. Because of its positively charged nature, it is also an innovative vector for the cell penetration of various compounds. We report a novel maurocalcine analog with improved properties: (i) the complete loss of pharmacological activity, (ii) preservation of the potent ability to carry cargo molecules into cells, and (iii) coupling chemistries not affected by the presence of internal cysteine residues of maurocalcine. We did this by replacing the six internal cysteine residues of maurocalcine by isosteric 2-aminobutyric acid residues and by adding an additional N-terminal biotinylated lysine (for a proof of concept analog) or an N-terminal cysteine residue (for a chemically competent coupling analogue). Additional replacement of a glutamate residue by alanyl at position 12 further improves the potency of these analogues. Coupling to several cargo molecules or nanoparticles are presented to illustrate the cell penetration potency and usefulness of these pharmacologically inactive analogs.

  20. Nanostructured Carbon Materials as Supports in the Preparation of Direct Methanol Fuel Cell Electrocatalysts

    Directory of Open Access Journals (Sweden)

    María Jesús Lázaro

    2013-08-01

    Full Text Available Different advanced nanostructured carbon materials, such as carbon nanocoils, carbon nanofibers, graphitized ordered mesoporous carbons and carbon xerogels, presenting interesting features such as high electrical conductivity and extensively developed porous structure were synthesized and used as supports in the preparation of electrocatalysts for direct methanol fuel cells (DMFCs. The main advantage of these supports is that their physical properties and surface chemistry can be tailored to adapt the carbonaceous material to the catalytic requirements. Moreover, all of them present a highly mesoporous structure, diminishing diffusion problems, and both graphitic character and surface area can be conveniently modified. In the present work, the influence of the particular features of each material on the catalytic activity and stability was analyzed. Results have been compared with those obtained for commercial catalysts supported on Vulcan XC-72R, Pt/C and PtRu/C (ETEK. Both a highly ordered graphitic and mesopore-enriched structure of these advanced nanostructured materials resulted in an improved electrochemical performance in comparison to the commercial catalysts assayed, both towards CO and alcohol oxidation.

  1. Preparation and characterization of dye-sensitized TiO{sub 2} nanorod solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Meng, Lijian, E-mail: ljm@isep.ipp.pt [Departamento de Física, Instituto Superior de Engenharia do Porto, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 431, 4200-072 Porto (Portugal); Centro de Física, Universidade do Minho, 4800-058 Guimarães (Portugal); Chen, Hong [Key Laboratory of Optical System Advanced Manufacturing Technology, Changchun Institute of Optics, fine Mechanics and Physics of Chinese Academy of Science, Changchun 130033 (China); Li, Can [State Key Laboratory of Catalysis, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023,China (China); Santos, M.P. dos [CEFITEC, Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Departamento de Física, Escola de Ciências e Tecnologia, Universidade de Évora (Portugal)

    2015-02-27

    TiO{sub 2} nanorods were prepared by DC reactive magnetron sputtering technique and applied to dye-sensitized solar cells (DSSCs). The length of the TiO{sub 2} nanorods was varied from 1 μm to 6 μm. The scanning electron microscopy images show that the nanorods are perpendicular to the substrate. Both the X-ray diffraction patterns and Raman scattering results show that the nanorods have an anatase phase; no other phase has been observed. (101) and the (220) diffraction peaks have been observed for the TiO{sub 2} nanorods. The (101) diffraction peak intensity remained constant despite the increase of nanorod length, while the intensity of the (220) diffraction peak increased almost linearly with the nanorod length. These nanorods were used as the working electrodes in DSSCs and the effect of the nanorod length on the conversion efficiency has been studied. An optimum photoelectric conversion efficiency of 4.8% has been achieved for 4 μm length nanorods. - Highlights: • [110] oriented TiO{sub 2} nanorods were deposited on ITO substrate by dc reactive magnetron sputtering. • The structural properties of these nanorods have been studied. • The (110) texture is dominated by strain energy minimization. • DSSCs were assembled using these nanorods as electrode.

  2. Preparation, in vitro mineralization and osteoblast cell response of electrospun 13-93 bioactive glass nanofibers.

    Science.gov (United States)

    Deliormanlı, Aylin M

    2015-08-01

    In this study, silicate based 13-93 bioactive glass fibers were prepared through sol-gel processing and electrospinning technique. A precursor solution containing poly (vinyl alcohol) and bioactive glass sol was used to produce fibers. The mixture was electrospun at a voltage of 20 kV by maintaining tip to a collector distance of 10 cm. The amorphous glass fibers with an average diameter of 464±95 nm were successfully obtained after calcination at 625 °C. Hydroxyapatite formation on calcined 13-93 fibers was investigated in simulated body fluid (SBF) using two different fiber concentrations (0.5 and 1 mg/ml) at 37 °C. When immersed in SBF, conversion to a calcium phosphate material showed a strong dependence on the fiber concentration. At 1mg/ml, the surface of the fibers converted to the hydroxyapatite-like material in SBF only after 30 days. At lower solid concentrations (0.5 mg/ml), an amorphous calcium phosphate layer formation was observed followed by the conversion to hydroxyapatite phase after 7 days of immersion. The XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) assay was conducted to evaluate the osteoblast cell response to the bioactive glass fibers. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Education catching up with science: preparing students for three-dimensional literacy in cell biology.

    Science.gov (United States)

    Kramer, Ijsbrand M; Dahmani, Hassen-Reda; Delouche, Pamina; Bidabe, Marissa; Schneeberger, Patricia

    2012-01-01

    The large number of experimentally determined molecular structures has led to the development of a new semiotic system in the life sciences, with increasing use of accurate molecular representations. To determine how this change impacts students' learning, we incorporated image tests into our introductory cell biology course. Groups of students used a single text dealing with signal transduction, which was supplemented with images made in one of three iconographic styles. Typically, we employed realistic renderings, using computer-generated Protein Data Bank (PDB) structures; realistic-schematic renderings, using shapes inspired by PDB structures; or schematic renderings, using simple geometric shapes to represent cellular components. The control group received a list of keywords. When students were asked to draw and describe the process in their own style and to reply to multiple-choice questions, the three iconographic approaches equally improved the overall outcome of the tests (relative to keywords). Students found the three approaches equally useful but, when asked to select a preferred style, they largely favored a realistic-schematic style. When students were asked to annotate "raw" realistic images, both keywords and schematic representations failed to prepare them for this task. We conclude that supplementary images facilitate the comprehension process and despite their visual clutter, realistic representations do not hinder learning in an introductory course.

  4. Enhanced performance of microbial fuel cell with in situ preparing dual graphene modified bioelectrode.

    Science.gov (United States)

    Chen, Junfeng; Hu, Yongyou; Tan, Xiaojun; Zhang, Lihua; Huang, Wantang; Sun, Jian

    2017-10-01

    This study proposed a three-step method to prepare dual graphene modified bioelectrode (D-GM-BE) by in situ microbial-induced reduction of GO and polarity reversion in microbial fuel cell (MFC). Both graphene modified bioanode (GM-BA) and biocathode (GM-BC) were of 3D graphene/biofilm architectures; the viability and thickness of microbial biofilm decreased compared with control bioelectrode (C-BE). The coulombic efficiency (CE) of GM-BA was 2.1 times of the control bioanode (C-BA), which demonstrated higher rate of substrates oxidation; the relationship between peak current and scan rates data meant that GM-BC was of higher efficiency of catalyzing oxygen reduction than the control biocathode (C-BC). The maximum power density obtained in D-GM-BE MFC was 122.4±6.9mWm(-2), the interfacial charge transfer resistance of GM-BA and GM-BC were decreased by 79% and 75.7%. The excellent electrochemical performance of D-GM-BE MFC was attributed to the enhanced extracellular electron transfer (EET) process and catalyzing oxygen reduction. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Low temperature preparation and fuel cell properties of rare earth doped barium cerate solid electrolytes

    Institute of Scientific and Technical Information of China (English)

    蒋凯; 何志奇; 孟建; 任玉芳; 苏锵

    1999-01-01

    The solid electrolytes, BaCe0.8 Ln0.2O2.9 (Ln: Gd, Sm, Eu), were prepared by the sol-gel method. XRD indicated that a pure orthorhombic phase was formed at 900℃. The synthesis temperature by the sol-gel method was about 600℃ lower than the high temperature solid phase reaction method, The electrical conductivity and impedance spectra were measured and the conduction mechanism was studied. The grain-boundary resistance of the solid electrolyte could be reduced or eliminated by the sol-gel method. The conductivity of BaCe0.8Gd0.2O2.9 is 7.87×10-2 S·cm-1 at 800℃. The open-circuit voltage of hydrogen-oxygen fuel cell using BaCe0.8 Gd0.2O2.9 as electrolyte was near to 1 V and its maximum power density was 30 mW·cm-2.

  6. The impact of preparation and support procedures for children with sickle cell disease undergoing MRI

    Energy Technology Data Exchange (ETDEWEB)

    Cejda, Katherine R. [St. Jude Children' s Research Hospital, Child Life Program, Memphis, TN (United States); Smeltzer, Matthew P. [St. Jude Children' s Research Hospital, Department of Biostatistics, Memphis, TN (United States); Hansbury, Eileen N. [Baylor International Hematology Center of Excellence and the Texas Children' s Center for Global Health, Houston, TX (United States); McCarville, Mary Elizabeth; Helton, Kathleen J. [St. Jude Children' s Research Hospital, Department of Radiological Sciences, Memphis, TN (United States); Hankins, Jane S. [St. Jude Children' s Research Hospital, Department of Hematology, Memphis, TN (United States)

    2012-10-15

    Children with sickle cell disease (SCD) often undergo MRI studies to assess brain injury or to quantify hepatic iron. MRI requires the child to lie motionless for 30-60 min, thus sedation/anesthesia might be used to facilitate successful completion of exams, but this poses additional risks for SCD patients. To improve children's ability to cope with MRI examinations and avoid sedation, our institution established preparation and support procedures (PSP). To investigate the impact of PSP in reducing the need for sedation during MRI exams among children with SCD. Data on successful completion of MRI testing were compared among 5- to 12-year-olds who underwent brain MRI or liver R2*MRI with or without receiving PSP. Seventy-one children with SCD (median age 9.85 years, range 5.57-12.99 years) underwent a brain MRI (n = 60) or liver R2*MRI (n = 11). Children who received PSP were more likely to complete an interpretable MRI exam than those who did not 30 of 33; 91% vs. 27 of 38; 71%, unadjusted OR = 4.1 (P = 0.04) and OR = 8.5 (P < 0.01) when adjusting for age. PSP can help young children with SCD complete clinically interpretable, nonsedated MRI exams, avoiding the risks of sedation/anesthesia. (orig.)

  7. Efficiency enhancement of perovskite solar cells by fabricating as-prepared film before sequential spin-coating procedure

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, Jiajia [Jiangsu Key Laboratory of Materials and Technology for Energy Conversion, College of material science and technology, Nanjing University of Aeronautics and Astronautics, Nanjing 211100 (China); Tao, Hai jun, E-mail: taohaijun@nuaa.edu.cn [Jiangsu Key Laboratory of Materials and Technology for Energy Conversion, College of material science and technology, Nanjing University of Aeronautics and Astronautics, Nanjing 211100 (China); Jiangsu Collaborative Innovation Center of Photovolatic Science and Engineering, Changzhou University, Changzhou 213164, Jiangsu (China); Chen, Shanlong; Tan, Bin [Jiangsu Key Laboratory of Materials and Technology for Energy Conversion, College of material science and technology, Nanjing University of Aeronautics and Astronautics, Nanjing 211100 (China); Zhou, Ning [Shanghai Electrochemical Energy Devices Research Center, School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai 200240 (China); Zhu, Lumin; Zhao, Yuan [Jiangsu Key Laboratory of Materials and Technology for Energy Conversion, College of material science and technology, Nanjing University of Aeronautics and Astronautics, Nanjing 211100 (China); Wang, Yuqiao [Jiangsu Optoelectronic Functional Materials and Engineering Laboratory, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189 (China); Tao, Jie [Jiangsu Key Laboratory of Materials and Technology for Energy Conversion, College of material science and technology, Nanjing University of Aeronautics and Astronautics, Nanjing 211100 (China)

    2016-05-15

    Graphical abstract: Schematic illustration of modified two-step spin-coating procedure for MAPbI{sub 3} perovskite thin films. - Highlights: • An as-prepared CH{sub 3}NH{sub 3}PbI{sub 3} and PbI{sub 2} film was introduced before the traditional two-step process. • Smooth morphology and trace amount of remaining PbI{sub 2} benefit the performance of solar cell. • The optimal as-prepared film introduced improves the efficiency of CH{sub 3}NH{sub 3}PbI{sub 3} solar cells from 9.11% to 11.16%. - Abstract: Sequential spin-coating procedure is a widely adopted strategy to prepare CH{sub 3}NH{sub 3}PbI{sub 3} on mesostructured TiO{sub 2} electrode for organolead halide perovskite-based solar cells. However, this method suffers from the rough surface and excessively residual PbI{sub 2} in the resulting perovskite film, deteriorating the device performance seriously. Herein, a facial modified sequential solution deposition method, by introducing an as-prepared CH{sub 3}NH{sub 3}PbI{sub 3} and PbI{sub 2} film before the traditional two-step process, was proposed to fabricate the perovskite-based solar cell with smooth morphology and trace amount of remaining PbI{sub 2}. The optimal as-prepared film introduced improves the efficiency of CH{sub 3}NH{sub 3}PbI{sub 3} solar cells from 9.11% to 11.16%. The enhancement of device performance can be attributed to the increased light absorption ability and decreased recombination rate of carriers in CH{sub 3}NH{sub 3}PbI{sub 3} absorber.

  8. Biochemistry and molecular biology of competence of plant cell differentiation and regeneration in vitro—a review

    National Research Council Canada - National Science Library

    Urmil Sethi; Sipra Guha-Mukherjee

    1990-01-01

    In spite of accumulation of voluminous literature on induction of cell division and differentiation in tissue culture, little information is available about the biochemical and molecular events that control...

  9. Quantitative TCR:pMHC Dissociation Rate Assessment by NTAmers Reveals Antimelanoma T Cell Repertoires Enriched for High Functional Competence.

    Science.gov (United States)

    Gannon, Philippe O; Wieckowski, Sébastien; Baumgaertner, Petra; Hebeisen, Michaël; Allard, Mathilde; Speiser, Daniel E; Rufer, Nathalie

    2015-07-01

    Experimental models demonstrated that therapeutic induction of CD8 T cell responses may offer protection against tumors or infectious diseases providing that T cells have sufficiently high TCR/CD8:pMHC avidity for efficient Ag recognition and consequently strong immune functions. However, comprehensive characterization of TCR/CD8:pMHC avidity in clinically relevant situations has remained elusive. In this study, using the novel NTA-His tag-containing multimer technology, we quantified the TCR:pMHC dissociation rates (koff) of tumor-specific vaccine-induced CD8 T cell clones (n = 139) derived from seven melanoma patients vaccinated with IFA, CpG, and the native/EAA or analog/ELA Melan-A(MART-1)(26-35) peptide, binding with low or high affinity to MHC, respectively. We observed substantial correlations between koff and Ca(2+) mobilization (p = 0.016) and target cell recognition (p tumor-reactive T cell clones derived from patients vaccinated with the low-affinity (native) peptide expressed slower koff rates than those derived from patients vaccinated with the high-affinity (analog) peptide (p tumor-specific T cells bearing TCRs with high TCR/CD8:pMHC avidity (p < 0.0001). Altogether, TCR:pMHC interaction kinetics correlated strongly with T cell functions. Our study demonstrates the feasibility and usefulness of TCR/CD8:pMHC avidity assessment by NTA-His tag-containing multimers of naturally occurring polyclonal T cell responses, which represents a strong asset for the development of immunotherapy. Copyright © 2015 by The American Association of Immunologists, Inc.

  10. Immunohistochemical evaluation of endothelial cell adhesion molecules in human dental pulp: effects of tooth preparation and adhesive application.

    Science.gov (United States)

    Bagis, Bora; Atilla, Pergin; Cakar, Nur; Hasanreisoglu, Ufuk

    2007-08-01

    Studies have demonstrated that restorative procedures can initiate pulpal inflammation. Adhesion molecules on endothelial cells mediate the leukocyte-endothelium interaction, which is the fundamental event of inflammation. The aim of this study was to evaluate possible changes in the endothelial cell adhesion molecules (CAMs) of human dental pulp with tooth preparation, and after the application of one-step self-etch adhesive. Twenty healthy human premolars and third molars scheduled to be extracted for orthodontic reasons were randomly assigned to four experimental groups. Group 1 involved sound intact teeth representing the controls. In group 2, teeth were prepared for full crown and extracted within 2h. Groups 3 and 4 comprised the teeth coated with one-step self-etch adhesive, iBond Gluma inside following the preparation and extracted after 24 and 48h, respectively. Tissue distribution and staining intensity of CAMs including E-selectin, P-selectin, ICAM-1, VCAM-1 and PECAM-1 was investigated in the pulp samples using monoclonal antibodies and the streptavidin-biotin-horse-radish immunoperoxidase procedure. The assessment of immunohistochemical reactions was performed by two independent observers using a semi-quantitative scale. All the CAMs evaluated were expressed by the healthy pulp tissues. Significant alterations in the distribution and staining intensity of CAMs were detected following tooth preparation. One-step self-etch adhesive tested in the present study induced inflammatory reactions in the pulp (Padhesive on prepared teeth had a potential to interfere with the inflammatory response.

  11. Preparation of Biotubes with vascular cells component by in vivo incubation using adipose-derived stromal cell-exuding multi-microporous molds.

    Science.gov (United States)

    Iwai, Ryosuke; Tsujinaka, Takahiro; Nakayama, Yasuhide

    2015-12-01

    Biotubes, prepared using in-body tissue architecture (IBTA) technology, have adequate mechanical properties and excellent biocompatibility for vascular grafts. However, they have thin walls, lack vascular constructing cells, and are composed of subcutaneous connective tissues consisting mainly of collagen and fibroblasts. This study aimed to prepare Biotubes with a vascular-like structure including an endothelial cell lining and a smooth muscle cell by IBTA using adipose-derived vascular stromal cell (ADSCs)-exuding specially designed multiporous tubes (outer diameter 5 mm, length 24 mm, pore size 500 μm, pore number 180, cell number/tube >3.0 × 10(6)). ADSCs were separated from rat subcutaneous fat, suspended in a Matrigel™ solution at 4 °C, and then filled into the tubes. After the tubes were embedded into dorsal subcutaneous pouches of the same rats for 2 weeks, robust Biotubes with a wall thickness of >600 μm were formed surrounding the tubes. The luminal layer of the obtained Biotubes was dominated by the cells positive for an endothelial marker. Almost the entire intima, with a thickness of about 400 μm, was occupied with cells positive for a smooth muscle marker. Both cells were derived from ADSCs. Biotube walls were constructed by fusing ADSC-derived vascular constructing cells exuded from the tubes and fibroblasts and collagen from the surrounding connective tissue. A robust Biotubes with vascular cells component, were formed after only 2 weeks of subcutaneous incubation of ADSCs-exuding multiporous tubes.

  12. GnRH receptor activation competes at a low level with growth signaling in stably transfected human breast cell lines

    Directory of Open Access Journals (Sweden)

    Morgan Kevin

    2011-11-01

    Full Text Available Abstract Background Gonadotrophin releasing hormone (GnRH analogs lower estrogen levels in pre-menopausal breast cancer patients. GnRH receptor (GnRH-R activation also directly inhibits the growth of certain cells. The applicability of GnRH anti-proliferation to breast cancer was therefore analyzed. Methods GnRH-R expression in 298 primary breast cancer samples was measured by quantitative immunofluorescence. Levels of functional GnRH-R in breast-derived cell lines were assessed using 125I-ligand binding and stimulation of 3H-inositol phosphate production. Elevated levels of GnRH-R were stably expressed in cells by transfection. Effects of receptor activation on in vitro cell growth were investigated in comparison with IGF-I and EGF receptor inhibition, and correlated with intracellular signaling using western blotting. Results GnRH-R immunoscoring was highest in hormone receptor (triple negative and grade 3 breast tumors. However prior to transfection, functional endogenous GnRH-R were undetectable in four commonly studied breast cancer cell lines (MCF-7, ZR-75-1, T47D and MDA-MB-231. After transfection with GnRH-R, high levels of cell surface GnRH-R were detected in SVCT and MDA-MB-231 clones while low-moderate levels of GnRH-R occurred in MCF-7 clones and ZR-75-1 clones. MCF-7 sub-clones with high levels of GnRH-R were isolated following hygromycin phosphotransferase transfection. High level cell surface GnRH-R enabled induction of high levels of 3H-inositol phosphate and modest growth-inhibition in SVCT cells. In contrast, growth of MCF-7, ZR-75-1 or MDA-MB-231 clones was unaffected by GnRH-R activation. Cell growth was inhibited by IGF-I or EGF receptor inhibitors. IGF-I receptor inhibitor lowered levels of p-ERK1/2 in MCF-7 clones. Washout of IGF-I receptor inhibitor resulted in transient hyper-elevation of p-ERK1/2, but co-addition of GnRH-R agonist did not alter the dynamics of ERK1/2 re-phosphorylation. Conclusions Breast cancers

  13. [Simplified preparation of test-red blood cells for ABO blood grouping in a laboratory in Madagascar].

    Science.gov (United States)

    Rasamiravaka, T; Andrianarivelo, A M; Ramarison, G; Rakoto-Alson, A O; Rasamindrakotroka, A

    2011-10-01

    To ensure self-sufficiency and lower costs associated with reagent red blood cells, some medical laboratories produce their own test-red blood cells for plasma ABO blood grouping. However, given the vital importance of blood goup testing, it is essential to verify the reliability of these cells. The purpose of this study was to assess the quality of laboratory-made ABO test-red blood cells. This study comparing house made and commercially available test-red blood cells was carried out at the Medical Biology Training and Research Laboratory in Madagascar. This laboratory is attended by people wishing to obtain their blood group card. In this population, no discrepancy was found between the red cell and plasma tests. Comparison of test-red blood cells with commercially available reagent red blood cells showed no difference in reactivity in the first four days of conservation. However a decrease in the reactivity of house made cells appeared on the 5th day. House made red blood cells are costless than commercially available reagent red blood cells mainly due to the simplified method of preparation. However, since laboratory-made cells progressivley lose antigenic reactivity quicly, production must be repeated regularly and good internal quality control is necessary to ensure reliability.

  14. Preparation of chitosan-plasmid DNA nanoparticles encoding interleukin-12 and their expression in CT-26 colon carcinoma cells.

    Science.gov (United States)

    Hallaj-Nezhadi, Somayeh; Valizadeh, Hadi; Dastmalchi, Siavoush; Baradaran, Behzad; Jalali, Mohammad Barzegar; Dobakhti, Faramarz; Lotfipour, Farzaneh

    2011-01-01

    Interleukin-12 (Il-12) as a cytokine has been proved to possess antitumor effects via stimulating the immune system. Non-viral gene delivery systems exhibit low toxicity and are easier to prepare compared to their viral counterparts. In this study, we aimed to prepare plasmid DNA loaded chitosan nanoparticles for expression of Il-12 and to evaluate their physicochemical characteristics, cytotoxicity and transfection efficiency in Murine CT-26 colon carcinoma cells. Nanoparticles were prepared using a complex coacervation process at different N/P ratios and characterized in terms of size, zeta potential, polydispersity index, morphology, encapsulation efficiency and polyplex formation. The cytotoxicities and transfection efficiencies of the prepared polyplexes were evaluated by MTT assay and ELISA (for hIL-12, p70), respectively. Size and zeta potential varied from 76.73 to 867.03 nm and between 5.68 and 16.77 mV, respectively. Strong attachment of the DNA to chitosan was observed after polyplex preparation. Encapsulation efficiencies were high (72.97-94.87%). The transfection efficiencies of the prepared complexes were obviously higher than those of naked pDNA when N/P ratios were between 16 and 60. Maximum level of phIL-12 expression was obtained at (N/P = 16) with mean particle size of 381.83±82.77 nm (polydispersity index=0.44) indicating the improved transfection of pUMVC3-hIL12 about 2.80 times compared to that of the naked pUMVC3-hIL12. Prepared polyplexes were nontoxic to CT-26 cells. Chitosan-DNA nanoparticles at N/P = 16 with minimal cytotoxicity, can be used as suitable candidate for Il-12 delivery. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.

  15. Nursing students' views of clinical competence assessment.

    Science.gov (United States)

    Bradshaw, Carmel; O'Connor, Maureen; Egan, Geraldine; Tierney, Katie; Butler, Mary Pat; Fahy, Anne; Tuohy, Dympna; Cassidy, Irene; Quillinan, Bernie; McNamara, Mary C

    This paper reports on some outcomes of a research study evaluating a new assessment framework of clinical competence used in undergraduate nursing programmes in the Mid West Region of Ireland. First, this paper presents both the strengths and weaknesses of the present model, as articulated by student nurses. Second, it generates a broader critical debate around the concept of competency assessment. The model of competence in question was developed by the Irish Nursing Board then elaborated on by the University of Limerick in partnership with local health service providers in 2002. Methodology involved a triangulated approach, comprising a series of focus group interviews with students (n=13) and preceptors (n=16) followed by a survey of students (n=232) and preceptors (n=837). Findings from the student focus groups are reported here. Themes identified using Burnard's (1991) framework for analysis are preparation for competency assessment, competency documentation, supporting assessment in practice, organisational and resource factors and the competency assessment structure and process. Results from this research have implications for refinement and revision of the present competency assessment framework, for student and staff preparation and for collaboration between stakeholders.

  16. Preparation and characterization of mono-sheet bipolar membranes by pre-irradiation grafting method for fuel cell applications

    Science.gov (United States)

    Guan, Yingjie; Fang, Jun; Fu, Tao; Zhou, Huili; Wang, Xin; Deng, Zixiang; Zhao, Jinbao

    2016-09-01

    A new method for the preparation of the mono-sheet bipolar membrane applied to fuel cells was developed based on the pre-irradiation grafting technology. A series of bipolar membranes were successfully prepared by simultaneously grafting of styrene onto one side of the poly(ethylene-co-tetrafluoroethylene) base film and 1-vinylimidazole onto the opposite side, followed by the sulfonation and alkylation, respectively. The chemical structures and microstructures of the prepared membranes were investigated by ATR-FTIR and SEM-EDS. The TGA measurements demonstrated the prepared bipolar membranes have reasonable thermal stability. The ion exchange capacity, water uptake and ionic conductivity of the membranes were also characterized. The H2/O2 single fuel cells using these membranes were evaluated and revealed a maximum power density of 107 mW cm-2 at 35 °C with unhumidified hydrogen and oxygen. The preliminary performances suggested the great prospect of these membranes in application of bipolar membrane fuel cells.

  17. Exposure of pig oocytes to PCBs during in vitro maturation: effects on developmental competence, cytoplasmic remodelling and communications with cumulus cells

    Directory of Open Access Journals (Sweden)

    TAL Brevini

    2009-06-01

    Full Text Available Polychlorinated biphenyls (PCBs are one of the most persistent and widespread groups of endocrine disrupting compounds in the ecosystem. These substances are present in sewage sludge that is spread in increasing amounts on arable land and pasture as fertilizer, and are ingested by farm animals with food and drinking water. This study investigated the effect of different PCB concentrations on pig oocyte in vitro maturation and developmental competence as well as examined the possible mechanisms involved. A concentration ranging from 0 to 1 ?g/mL of Aroclor 1254 (A1254, a pool of more than 60 PCB congeners, was added to the maturation medium, as its composition is considered environmentally relevant. A1254 had no effect on maturation of pig oocytes and on the number of oocytes that cleaved following parthenogenetic activation at any of the doses tested. By contrast, a significant decrease in the number of zygotes that developed to blastocyst stage became evident at a concentration of 10 ng/mL. The number of blastocysts obtained decreased significantly, and in a dose response manner with higher concentrations. Exposure to PCBs altered mitochondria relocation during maturation and this was associated with the lack of a cytoplasmic microtubule network. No effect on mitochondria activity was observed. A1254 exposure also perturbed gap-junction mediated communications between oocytes and cumulus cells. In conclusion, PCB exposure of pig oocytes during in vitro maturation significantly decreased oocyte developmental competence, altered both their cytoplasmic remodelling and the communication with the somatic compartment. These data indicated that accumulation of PCBs in the pig organism may have a detrimental effect on the reproductive efficiency in this species.

  18. Preparation of Silicon-Carbon-Based Dots@Dopamine and Its Application in Intracellular Ag(+) Detection and Cell Imaging.

    Science.gov (United States)

    Jiang, Yuliang; Wang, Zhaoyin; Dai, Zhihui

    2016-02-17

    A novel nanocomposite, silicon-carbon-based dots@dopamine (Si-CDs@DA) was prepared using (3-aminopropyl) triethoxysilane, glycerol, and dopamine as raw materials via a rapid microwave-assisted irradiation. This type of Si-CDs@DA exhibited ultrabright fluorescence emission (quantum yield of 12.4%) and could response to Ag(+) selectively and sensitively. Moreover, the obtained Si-CDs@DA can be further applied in sensing intracellular Ag(+) and cell imaging, because of its photostability, salt stability, and low cytotoxicity. This study provides a simple and efficient approach for preparing novel Ag(+) fluorescent probes, which could expand the application of carbon nanomaterials in designing related biosensors.

  19. In-Vitro Adhesion and Invasion Properties of Salmonella Typhimurium Competing with Bacteriophage in Epithelial Cells and Chicken Macrophages

    Directory of Open Access Journals (Sweden)

    HY Lee

    2015-12-01

    Full Text Available ABSTRACT This study was designed to assess the role of bacteriophage P22 in the adhesion, invasion, intracellular survival of, and cellular immune response to Salmonella Typhimurium in intestinal epithelial INT-407 and chicken macrophage-like HD11 cells. The ability of S. Typhimurium to adhere, invade, and survive to INT-407 and HD11cells was evaluated under Salmonella infection alone (control, phage treatment followed by Salmonella infection (PS, Salmonella infection followed by phage treatment (SP, and a combination treatment with Salmonella and phage (S+P. The number of S. Typhimurium associated on INT-407 cells was reduced from 4.2 to 2.7 log cfu/cm2 by phage treatment (SP. The number of intracellular S. Typhimurium within INT-407 cells was significantly reduced to below the detection limit (0.7 log cfu/cm2 compared with the control (3.4 log cfu/cm2. S. Typhimurium remained inside HD11 cells at 49% and 17% levels in the absence and presence of phages, respectively, at 24 h post-infection (hpi. The expression levels of IFN-g, IL-10, IL-1b, IL-6, IL-8, iNOS, and IL-12 increased in HD11 cells regardless the absence and presence of phages, while those of IL-16, TLR2-1, TLR3, and TLR7 were decreased at 0 and 24 hpi. This study sheds new light on our understanding of the role of phages in Salmonella adhesion, invasion, survival, and cellular immune responses.

  20. Assessment of Innovation Competency

    DEFF Research Database (Denmark)

    Nielsen, Jan Alexis

    2015-01-01

    of the recorded talk in interaction that occurred in teacher group discussion sessions at 5 upper secondary schools. Based on the analysis, it was possible to extrapolate assessment criteria for 5 subcompetencies relevant to innovation (creative competency, collaboration competency, navigation competency, action...... competency, and communication competency) as well as assessment criteria for a number of skills relevant to these subcompetencies. These assessment criteria, it is argued, largely resonate with existing literature and they provide a detailed glimpse into how assessment of innovation competency could...

  1. Optimization and comparison of two different 3D culture methods to prepare cell aggregates as a bioink for organ printing.

    Science.gov (United States)

    Imani, Rana; Hojjati Emami, Shahriar; Fakhrzadeh, Hossein; Baheiraei, Nafiseh; Sharifi, Ali M

    2012-04-01

    The ultimate goal of tissue engineering is to design and fabricate functional human tissues that are similar to natural cells and are capable of regeneration. Preparation of cell aggregates is one of the important steps in 3D tissue engineering technology, particularly in organ printing. Two simple methods, hanging drop (HD) and conical tube (CT) were utilized to prepare cell aggregates. The size and viability of the aggregates obtained at different initial cell densities and pre-culture duration were compared. The proliferative ability of the cell aggregates and their ability to spread in culture plates were also investigated. In both methods, the optimum average size of the aggregates was less than 500 microm. CT aggregates were smaller than HD aggregates. 5,000 cells per drop HD aggregates showed a marked ability to attach and spread on the culture surface. The proliferative ability reduced when the initial cell density was increased. Comparing these methods, we found that the HD method having better size controlling ability as well as enhanced ability to maintain higher rates of viability, spreading, and proliferation. In conclusion, smaller HD aggregates might be a suitable choice as building blocks for making bioink particles in bioprinting technique.

  2. Variations of secretome profiles according to conditioned medium preparation: The example of human mesenchymal stem cell-derived adipocytes.

    Science.gov (United States)

    Clabaut, Aline; Grare, Céline; Léger, Thibaut; Hardouin, Pierre; Broux, Odile

    2015-10-01

    One challenging point in analyzing cellular secretome collected as conditioned medium is cross-contamination by cell culture media components, especially bovine serum proteins. A common approach for serum removal is to wash the cells, an alternative is to grow cells using serum-free conditions. Given that the sample processing may influence the phenotype of cells and thus the secretome, it is important to establish the optimal protocol for each cell type. In this study, we compared two methods for preparing conditioned medium from human adipocytes derived from mesenchymal stem cells. Cells were either washed twice with PBS or cultured the last four days of differentiation in serum-free adipogenic medium. Gene expression of the cells was evaluated by using real-time PCR and 1D LC-MS/MS was used to compare secreted proteins present in the culture supernatants. Surprisingly, results showed significant differences in gene expression patterns of the cells and in protein content of the conditioned media and suggested that PBS washes induced severe modifications of the phenotype of cells and thus changes in protein secretion profiles. These data emphasize the significant variations in protein species related to cell manipulations and underline the importance of procedure optimization prior to any proteomic investigation.

  3. Young Investigator Challenge: A novel, simple method for cell block preparation, implementation, and use over 2 years.

    Science.gov (United States)

    Lindsey, Kathryn G; Houser, Patricia M; Shotsberger-Gray, Wanda; Chajewski, Olga S; Yang, Jack

    2016-12-01

    The cell block is an essential adjunct to conventional cytopreparatory techniques. The need for molecular analysis and immunostains will increase the need for successful cell block preparation. Even with this need, to the authors' knowledge very little has changed regarding the way in which cell blocks are produced. The authors developed A Formalin-Fixed, paraffin Embedded Cytology cell block Technique (AFFECT) that uses a cytospin centrifuge and funnel to deposit a cell pellet into a well on a piece of open-cell, absorbent foam. The foam and the pellet are then sent through normal processing. Herein, the authors present the implementation of this method and some of their experience with its performance over the course of 2 years. Although a comparison of the methods indicated good correlation for the production of a cell block between AFFECT and the agarose method, the AFFECT blocks demonstrated markedly improved cellular morphology. Over the first 6 months of use, AFFECT produced a successful cell block in 74% of cases overall, and in 65% of cases with a cell pellet measuring ≤0.1 mL. The year preceding the implementation of AFFECT and its first year of use were compared for endoscopic and bronchoscopic ultrasound-guided fine-needle aspiration specimens, and demonstrated an improved success rate. The authors developed a novel method of cell block preparation that demonstrates improved histology and has increased the success rate of cell block production compared with the agarose method. Cancer Cytopathol 2016;124:885-892. © 2016 American Cancer Society. © 2016 American Cancer Society.

  4. PREPARATION AND PROPERTIES OF SPAES-TiO_2 HYBRID MEMBRANES FOR DIRECT METHANOL FUEL CELL

    Institute of Scientific and Technical Information of China (English)

    Ni Zhang; Hui-ling Liu; Jun-jing Li; Zhi Xia

    2009-01-01

    Sulfonated poly(arylene ether sulfone) (SPAES) copolymer with degree of sulfonation of 1.0 was synthesized and characterized. A series of SPAES-TiO_2 hybrid membranes with various contents of nano-sized TiO_2 particles were prepared and characterized through sol-gel reactions. Scanning electron microscopy (SEM) images indicated the TiO_2 particles were well dispersed within polymer matrix. These composite membranes were evaluated for proton exchange membranes (PEMs) in direct methanol fuel cell (DMFC). These membranes showed good thermal stability and mechanical properties. It was found that the water uptake of these membranes increased with the increase of the TiO_2 contents in the hybrid membranes. Meanwhile, the introduction of inorganic particles increased the proton conductivity and reduced the methanol permeability. The proton conductivities (0.118-0.162 S/cm) of SPAES-TiO_2 hybrid membranes with 8% TiO_2 particles are much higher than those of Nation 117 membrane (0.095-0.117 S/cm) and pure SPAES membrane (0.100-0.124 S/cm) with degree of sulfonation of 1.0 at all temperatures (25-100℃). Especially, the methanol diffusion coefficient (8.4×10~(-7) cm~2/s) of SPAES-TiO_2 hybrid membranes with 8% TiO_2 is much lower than that of Nation 117 membrane (2.1 ×10~(-6) cm~2/s). SPAES-TiO_2 hybrid membranes were therefore proposed as candidates of material for PEM in DMFC.

  5. Preparation and characterization of TiO2 barrier layers for dye-sensitized solar cells.

    Science.gov (United States)

    Zheng, Yichen; Klankowski, Steven; Yang, Yiqun; Li, Jun

    2014-07-09

    A TiO2 barrier layer is critical in enhancing the performance of dye-sensitized solar cells (DSSCs). Two methods to prepare the TiO2 barrier layer on fluorine-doped tin dioxide (FTO) surface were systematically studied in order to minimize electron-hole recombination and electron backflow during photovoltaic processes of DSSCs. The film structure and materials properties were correlated with the photovoltaic characteristics and electrochemical properties. In the first approach, a porous TiO2 layer was deposited by wet chemical treatment of the sample with TiCl4 solution for time periods varying from 0 to 60 min. The N719 dye molecules were found to be able to insert into the porous barrier layers. The 20 min treatment formed a nonuniform but intact TiO2 layer of ∼100-300 nm in thickness, which gave the highest open-circuit voltage VOC, short-circuit photocurrent density JSC, and energy conversion efficiency. But thicker TiO2 barrier layers by this method caused a decrease in JSC, possibly limited by lower electrical conductance. In the second approach, a compact TiO2 barrier layer was created by sputter-coating 0-15 nm Ti metal films on FTO/glass and then oxidizing them into TiO2 with thermal treatment at 500 °C in the air for 30 min. The dye molecules were found to only attach at the outer surface of the barrier layer and slightly increased with the layer thickness. These two kinds of barrier layer showed different characteristics and may be tailored for different DSSC studies.

  6. Effervescent Granules Prepared Using Eucommia ulmoides Oliv. and Moso Bamboo Leaves: Hypoglycemic Activity in HepG2 Cells

    Directory of Open Access Journals (Sweden)

    Xiang-Zhou Li

    2016-01-01

    Full Text Available Eucommia ulmoides Oliv. (E. ulmoides Oliv. and moso bamboo (Phyllostachys pubescens leaves are used as folk medicines in central-western China to treat diabetes. To investigate the hypoglycemic activity of the effervescent granules prepared using E. ulmoides Oliv. and moso bamboo leaves (EBEG in HepG2 cells, EBEG were prepared with 5% of each of polysaccharides and chlorogenic acids from moso bamboo and E. ulmoides Oliv. leaves, respectively. HepG2 cells cultured in a high-glucose medium were classified into different groups. The results displayed EBEG-treated cells showed better glucose utilization than the negative controls; thus, the hypoglycemic effect of EBEG was much greater than that of granules prepared using either component alone, thereby indicating that this effect was due to a synergistic action of the components. Further, glucose consumption levels in the cells treated with EBEG (156.35% at 200 μg/mL and the positive controls (metformin, 162.29%; insulin, 161.52% were similar. Thus, EBEG exhibited good potential for use as a natural antidiabetic agent. The hypoglycemic effect of EBEG could be due to the synergistic action of polysaccharides from the moso bamboo leaves and chlorogenic acids from E. ulmoides Oliv. leaves via the inhibition of alpha-glucosidase and glucose-6-phosphate displacement enzyme.

  7. Preparation of flexible TiO2 photoelectrodes for dye-sensitized solar cells

    Science.gov (United States)

    Li, Wen-Ren; Wang, Hsiu-Hsuan; Lin, Chia-Feng; Su, Chaochin

    2014-09-01

    Dye-sensitized solar cells (DSSCs) based on nanocrystalline TiO2 photoelectrodes on indium tin oxide (ITO) coated polymer substrates have drawn great attention due to its lightweight, flexibility and advantages in commercial applications. However, the thermal instability of polymer substrates limits the process temperature to below 150 °C. In order to assure high and firm interparticle connection between TiO2 nanocrystals (TiO2-NC) and polymer substrates, the post-treatment of flexible TiO2 photoelectrodes (F-TiO2-PE) by mechanical compression was employed. In this work, Degussa P25 TiO2-NC was mixed with tert-butyl alcohol and DI-water to form TiO2 paste. F-TiO2-PE was then prepared by coating the TiO2 paste onto ITO coated polyethylene terephthalate (PET) substrate using doctor blade followed by low temperature sintering at 120 °C for 2 hours. To study the effect of mechanical compression, we applied 50 and 100 kg/cm2 pressure on TiO2/PET to complete the fabrication of F-TiO2-PE. The surface morphology of F-TiO2-PE was characterized using scanning electron microscopy. The resultant F-TiO2-PE sample exhibited a smooth, crack-free structure indicating the great improvement in the interparticle connection of TiO2-NC. Increase of compression pressure could lead to the increase of DSSC photoconversion efficiency. The best photoconversion efficiency of 4.19 % (open circuit voltage (Voc) = 0.79 V, short-circuit photocurrent density (Jsc) = 7.75 mA/cm2, fill factor (FF) = 0.68) was obtained for the F-TiO2-PE device, which showed great enhancement compared with the F-TiO2-PE cell without compression treatment. The effect of compression in DSSC performance was vindicated by the electrochemical impedance spectroscopy measurement.

  8. Preparation of immunomagnetic nanoparticles and their application in the separation of mouse CD34 + hematopoietic stem cells

    Science.gov (United States)

    Liang, Xin; Xu, Ke; Xu, Jianhe; Chen, Wei; Shen, Hebai; Liu, Jianwen

    2009-06-01

    The magnetic nanoparticles with a diameter of about 60 nm were synthesized by coprecipitation from ferrous and ferric iron solutions and coated with silica. Then the nanoparticles were modified with N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAPS) in order to immobilize anti-CD34 + monoclonal antibodies to the surface of modified magnetic particles. The results of transmission electron microscope (TEM) and Fourier transformed infrared (FT-IR) indicated that the nanoparticles were successfully prepared. Scanning electron microscope (SEM) photo confirmed that the mouse CD34 + cells (cells expressing CD34) were separated by the immunomagnetic nanoparticles. The viability of the separated cells was studied by hematopoietic colony-forming assay, the result of which showed that the target cells still had an ability of proliferation and differentiation. The application of the separated CD34 + cells was in testing the pharmacological effect of three samples isolated from enzyme-digested traditional Chinese medicine Colla corii asini.

  9. Characterization of CuInSe{sub 2}/CdS thin-film solar cells prepared using CBD

    Energy Technology Data Exchange (ETDEWEB)

    Vidyadharan Pillai, P.K.; Vijayakumar, K.P. [Department of Physics, Cochin University of Science and Technology, Kochi, Kerala (India)

    1997-12-19

    CuInSe{sub 2}/CdS thin-film heterojunction solar cells were fabricated entirely by chemical bath deposition technique. The illuminated J-V characteristics of the devices prepared with different thicknesses of CdS and CuInSe{sub 2} were studied. The typical solar cell parameters obtained for the best cell are: V{sub oc}=365 mV, J{sub sc}=12 mA/cm{sup 2}, FF=61%, and {eta}=3.1% under an illumination of 85 mW/cm{sup 2} on a cell of active area 0.1 cm{sup 2}. The J-V and C-V characteristics under dark condition and the spectral response were also studied for the best cell. The diode quality factor obtained is 1.7

  10. Preparation of microcrystalline single junction and amorphous-microcrystalline tandem silicon solar cells entirely by hot-wire CVD

    Energy Technology Data Exchange (ETDEWEB)

    Kupich, M.; Grunsky, D.; Kumar, P.; Schroeder, B. [University of Kaiserslautern (Germany). Department of Physics

    2004-01-25

    The hot-wire chemical vapour deposition (HWCVD) has been used to prepare highly conducting p- and n-doped microcrystalline silicon thin layers as well as highly photoconducting, low defect density intrinsic microcrystalline silicon films. These films were incorporated in all-HWCVD, all-microcrystalline nip and pin solar cells, achieving conversion efficiencies of {eta}=5.4% and 4.5%, respectively. At present, only the nip-structures are found to be stable against light-induced degradation. Furthermore, microcrystalline nip and pin structures have been successfully incorporated as bottom cells in all-hot-wire amorphous-microcrystalline nipnip- and pinpin-tandem solar cells for the first time. So far, the highest conversion efficiencies of the 'micromorph' tandem structures are {eta}=5.7% for pinpin-solar cells and 7.0% for nipnip solar cells. (author)

  11. Approach to a case of multiple irregular red cell antibodies in a liver transplant recipient: Need for developing competence

    Directory of Open Access Journals (Sweden)

    Ravi C Dara

    2015-01-01

    Full Text Available Liver transplant procedure acts as a challenge for transfusion services in terms of specialized blood components, serologic problems, and immunologic effects of transfusion. Red cell alloimmunization in patients awaiting a liver transplant complicate the process by undue delay or unavailability of compatible red blood cell units. Compatible blood units can be provided by well-equipped immunohematology laboratory, which has expertise in resolving these serological problems. This report illustrates resolution of a case with multiple alloantibodies using standard techniques, particularly rare antisera. Our case re-emphasizes the need for universal antibody screening in all patients as part of pretransfusion testing, which helps to identify atypical antibodies and plan for appropriate transfusion support well in time. We recommend that the centers, especially the ones that perform complex procedures like solid organ transplants and hematological transplants should have the necessary immunohematological reagents including rare antisera to resolve complex cases of multiple antibodies as illustrated in this case.

  12. Production of Alexa Fluor 488-labeled reovirus and characterization of target cell binding, competence, and immunogenicity of labeled virions.

    Science.gov (United States)

    Fecek, Ronald J; Busch, Ryan; Lin, Hong; Pal, Kasturi; Cunningham, Cynthia A; Cuff, Christopher F

    2006-07-31

    Respiratory enteric orphan virus (reovirus) has been used to study many aspects of the biology and genetics of viruses, viral infection, pathogenesis, and the immune response to virus infection. This report describes the functional activity of virus labeled with Alexa Fluor 488, a stable fluorescent dye. Matrix assisted laser desorption-time of flight analysis indicated that Alexa Fluor 488 labeled the outer capsid proteins of reovirus. Labeled virus bound to murine L929 fibroblasts as determined by flow cytometry and fluorescence microscopy, and the specificity of binding were demonstrated by competitive inhibition with non-labeled virus. Labeled reovirus induced apoptosis and cytopathic effect in infected L929 cells. Mice infected with labeled virus mounted robust serum antibody and CD8(+) T-cell responses, indicating that labeled virus retained immunogenicity in vivo. These results indicate that Alexa Fluor 488-labeled virus provides a powerful new tool to analyze reovirus infection in vitro and in vivo.

  13. The competing roles of i-ZnO in Cu(In,Ga)Se2 solar cells

    NARCIS (Netherlands)

    Williams, B.L.; Zardetto, V.; Kniknie, B.; Verheijen, M.A.; Kessels, W.M.M.; Creatore, M.

    2016-01-01

    The electrical role of the highly resistive and transparent (HRT) i-ZnO layer in Cu(In, Ga)Se2(CIGS) solar cells is investigated. By tuning the resistivity of atomic layer deposited (ALD) i-ZnO through the use of post-growth O2-plasma treatments, it is shown that low i-ZnO carrier densities (i.e. hi

  14. Competencies: A New Sector.

    Science.gov (United States)

    Brophy, Monica; Kiely, Tony

    2002-01-01

    Job analysis of managers in 42 Irish three-star hotels identified the following key management competencies and associated behavioral indicators. The results were used to develop a competency framework for management development. (Contains 29 references.) (SK)

  15. Developing mathematical modelling competence

    DEFF Research Database (Denmark)

    Blomhøj, Morten; Jensen, Tomas Højgaard

    2003-01-01

    In this paper we introduce the concept of mathematical modelling competence, by which we mean being able to carry through a whole mathematical modelling process in a certain context. Analysing the structure of this process, six sub-competences are identified. Mathematical modelling competence...... cannot be reduced to these six sub-competences, but they are necessary elements in the development of mathematical modelling competence. Experience from the development of a modelling course is used to illustrate how the different nature of the sub-competences can be used as a tool for finding...... the balance between different kinds of activities in a particular educational setting. Obstacles of social, cognitive and affective nature for the students' development of mathematical modelling competence are reported and discussed in relation to the sub-competences....

  16. Inhibition of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus

    Science.gov (United States)

    Chang, Hong-Bin; Chen, Bing-Huei

    2015-01-01

    The objectives of this study were to explore the inhibition mechanism of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus. In addition, human bronchus epithelial cell line BEAS-2B (normal cell) was selected for comparison. A high-performance liquid chromatography (HPLC) method was developed to separate and quantify the various curcuminoids in C. longa extract, including curcumin (1,714.5 μg/mL), demethoxycurcumin (1,147.4 μg/mL), and bisdemethoxycurcumin (190.2 μg/mL). A high-stability nanoemulsion composed of Tween 80, water, and curcuminoid extract was prepared, with mean particle size being 12.6 nm. The cell cycle was retarded at G2/M for both the curcuminoid extract and nanoemulsion treatments; however, the inhibition pathway may be different. H460 cells were more susceptible to apoptosis than A549 cells for both curcuminoid extract and nanoemulsion treatments. Growth of BEAS-2B remained unaffected for both the curcuminoid extract and nanoemulsion treatments, with a concentration range from 1 to 4 μg/mL. Also, the activities of caspase-3, caspase-8, and caspase-9 followed a dose-dependent increase for both A549 and H460 cells for both the treatments, accompanied by a dose-dependent increase in cytochrome C expression and a dose-dependent decrease in CDK1 expression. Interestingly, a dose-dependent increase in cyclin B expression was shown for A549 cells for both the treatments, while a reversed trend was found for H460 cells. Both mitochondria and death receptor pathways may be responsible for apoptosis of both A549 and H460 cells. PMID:26345201

  17. Inhibition of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus.

    Science.gov (United States)

    Chang, Hong-Bin; Chen, Bing-Huei

    2015-01-01

    The objectives of this study were to explore the inhibition mechanism of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus. In addition, human bronchus epithelial cell line BEAS-2B (normal cell) was selected for comparison. A high-performance liquid chromatography (HPLC) method was developed to separate and quantify the various curcuminoids in C. longa extract, including curcumin (1,714.5 μg/mL), demethoxycurcumin (1,147.4 μg/mL), and bisdemethoxycurcumin (190.2 μg/mL). A high-stability nanoemulsion composed of Tween 80, water, and curcuminoid extract was prepared, with mean particle size being 12.6 nm. The cell cycle was retarded at G2/M for both the curcuminoid extract and nanoemulsion treatments; however, the inhibition pathway may be different. H460 cells were more susceptible to apoptosis than A549 cells for both curcuminoid extract and nanoemulsion treatments. Growth of BEAS-2B remained unaffected for both the curcuminoid extract and nanoemulsion treatments, with a concentration range from 1 to 4 μg/mL. Also, the activities of caspase-3, caspase-8, and caspase-9 followed a dose-dependent increase for both A549 and H460 cells for both the treatments, accompanied by a dose-dependent increase in cytochrome C expression and a dose-dependent decrease in CDK1 expression. Interestingly, a dose-dependent increase in cyclin B expression was shown for A549 cells for both the treatments, while a reversed trend was found for H460 cells. Both mitochondria and death receptor pathways may be responsible for apoptosis of both A549 and H460 cells.

  18. Preparation of nano-hydroxyapatite particles with different morphology and their response to highly malignant melanoma cells in vitro

    Science.gov (United States)

    Li, Bo; Guo, Bo; Fan, Hongsong; Zhang, Xingdong

    2008-11-01

    To investigate the effects of nano-hydroxyapatite (HA) particles with different morphology on highly malignant melanoma cells, three kinds of HA particles with different morphology were synthesized and co-cultured with highly malignant melanoma cells using phosphate-buffered saline (PBS) as control. A precipitation method with or without citric acid addition as surfactant was used to produce rod-like hydroxyapatite (HA) particles with nano- and micron size, respectively, and a novel oil-in-water emulsion method was employed to prepare ellipse-like nano-HA particles. Particle morphology and size distribution of the as prepared HA powders were characterized by transmission electron microscope (TEM) and dynamic light scattering technique. The nano- and micron HA particles with different morphology were co-cultured with highly malignant melanoma cells. Immunofluorescence analysis and MTT assay were employed to evaluate morphological change of nucleolus and proliferation of tumour cells, respectively. To compare the effects of HA particles on cell response, the PBS without HA particles was used as control. The experiment results indicated that particle nanoscale effect rather than particle morphology of HA was more effective for the inhibition on highly malignant melanoma cells proliferation.

  19. Preparation of nano-hydroxyapatite particles with different morphology and their response to highly malignant melanoma cells in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Li Bo [National Engineering Research Center for Biomaterial, Sichuan University, Chengdu 610064 (China); Guo Bo [National Engineering Research Center for Biomaterial, Sichuan University, Chengdu 610064 (China); West China Eye Center of Huaxi Hospital, Sichuan University, Chengdu 610064 (China); Fan Hongsong [National Engineering Research Center for Biomaterial, Sichuan University, Chengdu 610064 (China)], E-mail: leewave@126.com; Zhang Xingdong [National Engineering Research Center for Biomaterial, Sichuan University, Chengdu 610064 (China)

    2008-11-15

    To investigate the effects of nano-hydroxyapatite (HA) particles with different morphology on highly malignant melanoma cells, three kinds of HA particles with different morphology were synthesized and co-cultured with highly malignant melanoma cells using phosphate-buffered saline (PBS) as control. A precipitation method with or without citric acid addition as surfactant was used to produce rod-like hydroxyapatite (HA) particles with nano- and micron size, respectively, and a novel oil-in-water emulsion method was employed to prepare ellipse-like nano-HA particles. Particle morphology and size distribution of the as prepared HA powders were characterized by transmission electron microscope (TEM) and dynamic light scattering technique. The nano- and micron HA particles with different morphology were co-cultured with highly malignant melanoma cells. Immunofluorescence analysis and MTT assay were employed to evaluate morphological change of nucleolus and proliferation of tumour cells, respectively. To compare the effects of HA particles on cell response, the PBS without HA particles was used as control. The experiment results indicated that particle nanoscale effect rather than particle morphology of HA was more effective for the inhibition on highly malignant melanoma cells proliferation.

  20. Sample Preparation Methods Following CellSearch Approach Compatible of Single-Cell Whole-Genome Amplification: An Overview

    NARCIS (Netherlands)

    Swennenhuis, J.F.; Terstappen, L.W.M.M.; Kroneis, Thomas

    2015-01-01

    Single cells are increasingly used to determine the heterogeneity of therapy targets in the genome during the course of a disease. The first challenge using single cells is to isolate these cells from the surrounding cells, especially when the targeted cells are rare. A number of techniques have bee

  1. Preparation of 3D electrode microarrays of multi-walled carbon nanotubes/nafion nanocomposites for microfluidic biofuel cells.

    Science.gov (United States)

    Choi, Jin Ho; Kim, Young Ho; Choi, Sung Deuk; Kim, Gyu Man

    2014-12-01

    Three-dimensional (3D) electrode microarrays with multi-walled carbon nanotubes (MWCNTs) reinforced Nafion nanocomposites were prepared for microfluidic biofuel cells. The oxidized MWCNTs (ox-MWCNTs) were prepared using chemical reactions with 60% nitric acid solution with pristine MWCNTs at 120 degrees C for 12 hrs with a nitrogen gas flow environment. Ox-MWCNTs in the range of 1 to 20 wt.% based on the Nafion polymer weight were reinforced to Nafion nanocomposites by solution casting. The micro-porous structure of the ox-MWCNTs reinforced Nafion nanocomposites was prepared by plasma etching for 5 to 20 min. The 10 wt.% ox-MWCNTs reinforced Nafion nanocomposite produced stable micro-porous structures of 3D electrodes by 10 min plasma etching. Micro-scale 3D structures of MWCNTs reinforced Nafion nanocomposites in a diameter range of 47 to 300 μm were prepared by the micro-stencil assisted casting. To characterize the 3D electrode microarrays, the physical geometry and the reinforced MWCNT dispersion in the nanocomposite structure were examined using a scanning electron microscope (SEM) and an optical microscope. Thermal property measurements of the ox-MWCNTs reinforced Nafion nanocomposites with 10 min of plasma etching, and without plasma etching were made. Both showed stable thermal properties over 300 degrees C. The proposed 3D electrode microarray of MWCNT/Nafion nanocomposites with micro-porous structures can be applied to miniaturized fuel cell devices.

  2. Nanostructured TiO2 films for dye-sensitized solar cells prepared by the sol-gel method.

    Science.gov (United States)

    Jin, Young Sam; Kim, Kyung Hwan; Park, Sang Joon; Yoon, Hyon Hee; Choi, Hyung Wook

    2011-12-01

    TiO2 films were prepared on glass substrates using the sol-gel process for a dye-sensitized solar cell application. The TiO2 sol was prepared using hydrolysis/polycondensation. Titanium (IV) Tetra Isopropoxide (TTIP) was used as precursor and Nitric acid (HNO3) was used as a catalyst for the peptization. The crystal structure and morphology of the prepared materials were characterized by XRD, and an SEM. The observations confirmed the nanocrystalline nature of the TiO2. The reaction parameters, such as the catalyst concentrations, the calcination time, and the calcination temperature were varied during the synthesis in order to achieve nanosize TiO2 particles. The prepared TiO2 particles were coated onto FTO glass using a screen printing technique. The prepared TiO2 films were characterized by UV-vis. The TiO2 particles calcinated at low temperatures showed an anatase phase they grew into a rutile phase when the calcination temperature increased. The size and structure of the TiO2 particles were adjusted to specific surface areas. It was found that the conversion efficiency of the DSSC was highly affected by the properties of the TiO2 particles.

  3. Building Project Competence

    DEFF Research Database (Denmark)

    Pemsel, Sofia; Wiewiora, Anna

    This research investigates the development of project competence, and particularly, three related dynamic capabilities (shifting, adapting, leveraging) that contribute to project competence development. In doing so, we make use of the emerging literature on knowledge governance and theorize how k...... of dynamic capability building promoting project competence development....

  4. Identification of a novel MTOR activator and discovery of a competing endogenous RNA regulating autophagy in vascular endothelial cells.

    Science.gov (United States)

    Ge, Di; Han, Lei; Huang, ShuYa; Peng, Nan; Wang, PengChong; Jiang, Zheng; Zhao, Jing; Su, Le; Zhang, ShangLi; Zhang, Yun; Kung, HsiangFu; Zhao, BaoXiang; Miao, JunYing

    2014-06-01

    MTOR, a central regulator of autophagy, is involved in cancer and cardiovascular and neurological diseases. Modulating the MTOR signaling balance could be of great significance for numerous diseases. No chemical activators of MTOR have been found, and the urgent challenge is to find novel MTOR downstream components. In previous studies, we found a chemical small molecule, 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2(3H)-one (3BDO), that inhibited autophagy in human umbilical vein endothelial cells (HUVECs) and neuronal cells. Here, we found that 3BDO activated MTOR by targeting FKBP1A (FK506-binding protein 1A, 12 kDa). We next used 3BDO to detect novel factors downstream of the MTOR signaling pathway. Activation of MTOR by 3BDO increased the phosphorylation of TIA1 (TIA1 cytotoxic granule-associated RNA binding protein/T-cell-restricted intracellular antigen-1). Finally, we used gene microarray, RNA interference, RNA-ChIP assay, bioinformatics, luciferase reporter assay, and other assays and found that 3BDO greatly decreased the level of a long noncoding RNA (lncRNA) derived from the 3' untranslated region (3'UTR) of TGFB2, known as FLJ11812. TIA1 was responsible for processing FLJ11812. Further experiments results showed that FLJ11812 could bind with MIR4459 targeting ATG13 (autophagy-related 13), and ATG13 protein level was decreased along with 3BDO-decreased FLJ11812 level. Here, we provide a new activator of MTOR, and our findings highlight the role of the lncRNA in autophagy.

  5. Cu2ZnSnS4 solar cells prepared with sulphurized dc-sputtered stacked metallic precursors

    OpenAIRE

    Fernandes, P. A.; Salomé, P M P; Cunha, A. F. da; Schubert, Björn-Arvid

    2010-01-01

    In the present work we report the details of the preparation and characterization results of Cu2ZnSnS4 (CZTS) based solar cells. The CZTS absorber was obtained by sulphurization of dc magnetron sputtered Zn/Sn/Cu precursor layers. The morphology, composition and structure of the absorber layer were studied by scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction and Raman scattering. The majority carrier type was identified via a hot point probe analysis....

  6. Human IDO-competent, long-lived immunoregulatory dendritic cells induced by intracellular pathogen, and their fate in humanized mice

    Science.gov (United States)

    Tyagi, Rajeev K.; Miles, Brodie; Parmar, Rajesh; Garg, Neeraj K.; Dalai, Sarat K.; Baban, Babak; Cutler, Christopher W.

    2017-01-01

    Targeting of myeloid-dendritic cell receptor DC-SIGN by numerous chronic infectious agents, including Porphyromonas gingivalis, is shown to drive-differentiation of monocytes into dysfunctional mDCs. These mDCs exhibit alterations of their fine-tuned homeostatic function and contribute to dysregulated immune-responses. Here, we utilize P. gingivalis mutant strains to show that pathogen-differentiated mDCs from primary human-monocytes display anti-apoptotic profile, exhibited by elevated phosphorylated-Foxo1, phosphorylated-Akt1, and decreased Bim-expression. This results in an overall inhibition of DC-apoptosis. Direct stimulation of complex component CD40 on DCs leads to activation of Akt1, suggesting CD40 involvement in anti-apoptotic effects observed. Further, these DCs drove dampened CD8+ T-cell and Th1/Th17 effector-responses while inducing CD25+Foxp3+CD127− Tregs. In vitro Treg induction was mediated by DC expression of indoleamine 2,3-dioxygenase, and was confirmed in IDO-KO mouse model. Pathogen-infected & CMFDA-labeled MoDCs long-lasting survival was confirmed in a huMoDC reconstituted humanized mice. In conclusion, our data implicate PDDCs as an important target for resolution of chronic infection. PMID:28198424

  7. Effect of TiH2 on preparation of closed-cell aluminum foam and its compressive behavior

    Institute of Scientific and Technical Information of China (English)

    YANG Guo-jun; YU Hai-jun; YAO Guang-chun

    2006-01-01

    The vesicant problem during the process of preparing closed-cell aluminum foam by molten body transitional foaming process was discussed and the effect of granularity and addition of TiH2 on porosity of closed-cell aluminum foam was investigated.The static compressive behavior of closed-cell aluminum foam and the influence of porosity on static compressive property of closed-cell aluminum foam were researched as well. The results show that with increasing granularity of TiH2, the porosity of closed-cell aluminum foam firstly increases and then decreases gradually, the granularity should be controlled in the range of 38-74 μm which can result in higher porosity. The porosity of closed-cell aluminum foam increases with the increasing addition of TiH2,and the addition of TiH2 should be controlled from 1.5% to 2.5% which can result in homogeneous cell and moderate strength of closed-cell aluminum foam. The compressive process of closed-cell aluminum foam obviously displays linear elastic phase, plastic collapse phase, and densification phase, and the compressive strength grows with decreasing porosity.

  8. Development of a lung slice preparation for recording ion channel activity in alveolar epithelial type I cells

    Directory of Open Access Journals (Sweden)

    Crandall Edward D

    2005-04-01

    Full Text Available Abstract Background Lung fluid balance in the healthy lung is dependent upon finely regulated vectorial transport of ions across the alveolar epithelium. Classically, the cellular locus of the major ion transport processes has been widely accepted to be the alveolar type II cell. Although evidence is now emerging to suggest that the alveolar type I cell might significantly contribute to the overall ion and fluid homeostasis of the lung, direct assessment of functional ion channels in type I cells has remained elusive. Methods Here we describe a development of a lung slice preparation that has allowed positive identification of alveolar type I cells within an intact and viable alveolar epithelium using living cell immunohistochemistry. Results This technique has allowed, for the first time, single ion channels of identified alveolar type I cells to be recorded using the cell-attached configuration of the patch-clamp technique. Conclusion This exciting new development should facilitate the ascription of function to alveolar type I cells and allow us to integrate this cell type into the general model of alveolar ion and fluid balance in health and disease.

  9. Hysteretic Behavior upon Light Soaking in Perovskite Solar Cells Prepared via Modified Vapor-Assisted Solution Process.

    Science.gov (United States)

    Liu, Chong; Fan, Jiandong; Zhang, Xing; Shen, Yanjiao; Yang, Lin; Mai, Yaohua

    2015-05-06

    Recently, the organic-inorganic hybrid perovskite solar cells exhibit rapidly rising efficiencies, while anomalous hysteresis in perovskite solar cells remains unsolvable. Herein, a high-quality perovskite thin film is prepared by a modified vapor-assisted solution process, which is a simple but well-controllable method proven to be capable of producing a thin film with full surface coverage and grain size up to micrometers. The as-fabricated perovskite solar cell has efficiency as high as 10.2%. The hysteresis effects of both planar and mesoscopic TiO2-based perovskite solar cells have been comprehensively studied upon illumination. The results demonstrate that mesoporous-based perovskite cells combined with remarkable grain size are subject to alleviating the hysteresis effects in comparison to the planar cells. Likewise, mesoscopic TiO2-based perovskite cells perform independently of illumination and bias conditions prior to the measurements, whereas the planar cells display a reversible behavior of illumination and applied bias-dependent I-V curves. The present study would refer strip road for the stability study of the perovskite solar cells.

  10. Application of liquid-based cytology preparation in micronucleus assay of exfoliated buccal epithelial cells in road construction workers

    Directory of Open Access Journals (Sweden)

    P Arul

    2017-01-01

    Full Text Available Background: Asphalts are bitumens that consist of complex of hydrocarbon mixtures and it is used mainly in road construction and maintenance. Aim: This study was undertaken to evaluate the micronucleus (MN assay of exfoliated buccal epithelial cells in road construction workers using liquid-based cytology (LBC preparation. Materials and Methods: Three different stains (May–Grunwald Giemsa, hematoxylin and eosin, and Papanicolaou were used to evaluate the frequency of MN in exfoliated buccal epithelial of 100 participants (fifty road construction workers and fifty administrative staff using LBC preparation. Statistical analysis was performed with Student's t-test, and P< 0.05 was considered statistically significant. Results: The mean frequency of MN for cases was significantly higher than that of controls (P = 0.001 regardless of staining method used and also cases with exposure period of more than 5 years had statistically significant difference (P < 0.05 than cases with < 5 years of exposure. Conclusion: The present study concluded that workers exposed to asphalts during road construction exhibit a higher frequency of MN in exfoliated buccal epithelial cells and they are under the significant risk of cytogenetic damage. LBC preparation has potential application for the evaluation of frequency of MN. This technique may be advocated in those who are occupationally exposed to potentially carcinogenic agents in view of improvement in the smear quality and visualization of cell morphology.

  11. Quaternized chitosan-coated nanofibrous materials containing gossypol: preparation by electrospinning, characterization and antiproliferative activity towards HeLa cells.

    Science.gov (United States)

    Ignatova, Milena; Manolova, Nevena; Toshkova, Reneta; Rashkov, Iliya; Gardeva, Elena; Yossifova, Liliya; Alexandrov, Marin

    2012-10-15

    Nanofibrous polylactide-based materials loaded with a natural polyphenolic compound gossypol (GOS) with antitumor properties were prepared by electrospinning. The nanofibrous materials were coated with a thin film of crosslinked quaternized chitosan (QCh). GOS incorporated in the nanofibrous mats was in the amorphous state. GOS release was diffusion-controlled and its in vitro release profiles depended on the mat composition. The nanofibrous materials exhibited high cytotoxicity towards HeLa tumor cells. Interestingly, it was particularly pronounced in the case of fibrous materials, which contain both QCh and GOS. The observed strong antiproliferative effect of the nanofibrous mats was mainly due to induction of cell apoptosis.

  12. Herbal preparation (HemoHIM) enhanced functional maturation of bone marrow-derived dendritic cells mediated toll-like receptor 4

    OpenAIRE

    Lee, Sung-Ju; Kim, Jong-Jin; Kang, Kyung-Yun; Hwang, Yun-Ho; Jeong, Gil-Yeon; Jo, Sung-Kee; Jung, Uhee; Park, Hae-Ran; Yee, Sung-Tae

    2016-01-01

    Background HemoHIM, which is an herbal preparation of three edible herbs (Angelicam gigas Nakai, Cnidium offinale Makino, and Peaonia japonica Miyabe), is known to have various biological and immunological activities, but the modulatory effects of this preparation on dendritic cells (DCs)-mediated immune responses have not been examined previously. DCs are a unique group of white blood cells that initiate primary immune responses by capturing, processing, and presenting antigens to T cells. R...

  13. Herbal preparation (HemoHIM) enhanced functional maturation of bone marrow-derived dendritic cells mediated toll-like receptor 4

    OpenAIRE

    Lee, Sung-Ju; Kim, Jong-Jin; Kang, Kyung-Yun; Hwang, Yun-Ho; Jeong, Gil-Yeon; Jo, Sung-Kee; Jung, Uhee; Park, Hae-Ran; Yee, Sung-Tae

    2016-01-01

    Background HemoHIM, which is an herbal preparation of three edible herbs (Angelicam gigas Nakai, Cnidium offinale Makino, and Peaonia japonica Miyabe), is known to have various biological and immunological activities, but the modulatory effects of this preparation on dendritic cells (DCs)-mediated immune responses have not been examined previously. DCs are a unique group of white blood cells that initiate primary immune responses by capturing, processing, and presenting antigens to T cells. R...

  14. Effects of Yimu Shenghuasan Preparation on the Cytochrome P450 in Endometrial Cells and Immune Function of Dairy Cows

    Institute of Scientific and Technical Information of China (English)

    DU Jin-liang; QIN Jian-hua; CHU Jing-sheng; XU Li-na; MA Yu-zhong

    2010-01-01

    In order to investigate the mode of action of Yimu Shenghuasan preparation in endometrial cells of dairy cows,the primary cultured endometrial cells in cows were isolated and the inflammatory models were made by lipopolysaccharide(LPS)induction.The inflammatory cells were treated with gradient concentration of herbal medicine preparation,Yimu Shenghuasan for 48 and 72 h.The expression of cytochrome P450(CYP450)was detected by Western blot.The amounts of IgG and IgA in sera were also detected in the endometritis of dairy cows.The expression level of CYP450 in the endometrial cells of dairy cow was increased gradually,and the amounts of IgG,IgA were increased significantly as compared with those in the control group.The expression level of CYP450 in the inflammatory cells was increased significantly in the treatment of 2 000 μg mL-1 of Yimu Shenghuasan after 48 h of treatment.

  15. Promotion of glucose utilization by insulin enhances granulosa cell proliferation and developmental competence of porcine oocyte grown in vitro.

    Science.gov (United States)

    Itami, Nobuhiko; Munakata, Yasuhisa; Shirasuna, Koumei; Kuwayama, Takehito; Iwata, Hisataka

    2017-02-01

    In vitro culture of the oocyte granulosa cell complexes (OGCs) from early antral follicles (EAFs) shows granulosa cell (GC) proliferation, but to a lesser extent than that observed in vivo during follicle development. As the number of GCs closely relates to energy sufficiency of the oocytes, enhancement of GC proliferation influences oocyte development. GC proliferation depends on glycolysis and insulin-mediated AKT/mTOR signaling pathway; therefore, addition of culture medium containing insulin and glucose may potentially promote GC proliferation and hence improve oocyte development. In the present study, we assessed the effect of exogenous insulin and glucose concentration on GC proliferation and oocyte energy status as well as developmental abilities of porcine oocytes grown in vitro. In the presence of 5.5 mM of glucose (Low), a comparison of 10 versus 20 μg/ml insulin showed that high insulin enhanced GC proliferation but exhausted glucose from the medium, which resulted in low energy status including lipid and adenosine triphosphate of the oocyte. Whereas, in the presence of 20 μg/ml insulin, medium with 11 mM glucose (High) enhanced GC proliferation and oocyte energy status as well as developmental ability up to the blastocyst stage. Considering that there was no difference in OGCs development observed with medium (10 μg/ml insulin) containing 5.5 versus 11 mM glucose, we concluded that the combination of high insulin and glucose enhanced GC proliferation and energy status of oocytes as well as the developmental ability of the oocytes grown in vitro.

  16. Aqueous phase preparation of ultrasmall MoSe2 nanodots for efficient photothermal therapy of cancer cells

    Science.gov (United States)

    Yuwen, Lihui; Zhou, Jiajia; Zhang, Yuqian; Zhang, Qi; Shan, Jingyang; Luo, Zhimin; Weng, Lixing; Teng, Zhaogang; Wang, Lianhui

    2016-01-01

    Photothermal therapy (PTT) is a promising cancer treatment with both high effectiveness and fewer side effects. However, an ideal PTT agent not only needs strong absorption of near-infrared (NIR) light and high photothermal conversion efficiency, but also needs good biocompatibility, stability, and small size, which makes the design and preparation of a novel PTT agent a great challenge. In this work, we developed an ultrasonication-assisted liquid exfoliation method for the direct preparation of ultrasmall (2-3 nm) MoSe2 nanodots (NDs) in aqueous solution and demonstrated their superior properties as a PTT agent. The as-prepared MoSe2 NDs have strong absorption of NIR light and high photothermal conversion efficiency of about 46.5%. In vitro cellular experiments demonstrate that MoSe2 NDs have negligible cytotoxicity and can efficiently kill HeLa cells (human cervical cell line) under NIR laser (785 nm) irradiation.Photothermal therapy (PTT) is a promising cancer treatment with both high effectiveness and fewer side effects. However, an ideal PTT agent not only needs strong absorption of near-infrared (NIR) light and high photothermal conversion efficiency, but also needs good biocompatibility, stability, and small size, which makes the design and preparation of a novel PTT agent a great challenge. In this work, we developed an ultrasonication-assisted liquid exfoliation method for the direct preparation of ultrasmall (2-3 nm) MoSe2 nanodots (NDs) in aqueous solution and demonstrated their superior properties as a PTT agent. The as-prepared MoSe2 NDs have strong absorption of NIR light and high photothermal conversion efficiency of about 46.5%. In vitro cellular experiments demonstrate that MoSe2 NDs have negligible cytotoxicity and can efficiently kill HeLa cells (human cervical cell line) under NIR laser (785 nm) irradiation. Electronic supplementary information (ESI) available: Characterization, size distribution and EDS spectrum of MoSe2 NDs, calculation of

  17. Evidence of clinical competence.

    Science.gov (United States)

    Lejonqvist, Gun-Britt; Eriksson, Katie; Meretoja, Riitta

    2012-06-01

    This cross-sectional research used a qualitative questionnaire to explore clinical competence in nursing. The aim was to look for evidence of how clinical competence showed itself in practice. In the research, the views from both education and working life are combined to broadly explore and describe clinical competence from the perspective of students, clinical preceptors and teachers. The questions were formulated on how clinical competence is characterised and experienced, what contributes to it and how it is maintained, and on the relation between clinical competence and evidence-based care. The answers were analysed by inductive content analysis. The results showed that clinical competence in practice is encountering, knowing, performing, maturing and improving. Clinical competence is an ongoing process, rather than a state and manifests itself in an ontological and a contextual dimension.

  18. Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models

    OpenAIRE

    Ahlf Wheatcraft, Dorothy R.; Liu, Xin; Hummon, Amanda B.

    2014-01-01

    Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with some of the spatial and molecular complexity of tissue. For example, many cell lines form 3D structures given appropriate in vitro conditions. Colon cancer cell lines form 3D cell culture spheroids, in vitro mimics of avascular tumor nodules. While immunohistochemistry and other classical imaging methods are popular for monitoring the distribu...

  19. Antiproliferative activity of extracts prepared from three species of Reishi on cultured human normal and tumor cell lines.

    Science.gov (United States)

    Katagata, Yohtaro; Sasaki, Fumiyuki

    2010-01-01

    The present study investigated the growth of human fibrosarcoma (HT-1080) and fibroblast (SF-TY) cells in combination with water-soluble (WS) and high molecular component (HMC) fractions prepared from Reishi (R), Rokkaku-Reishi (2R) and Apple Rokkaku-Reishi (A2R). Each WS fraction exhibited dose-and time-dependent inhibition of the growth of the HT-1080 and SF-TY cells. The extracts exhibited marked antiproliferative activity against the HT-1080 cells. The HMC fractions inhibited cell growth dose-and time-dependently in the HT-1080 cells only, and not in the SF-TY cells, suggesting that HMC fractions selectively inhibit HT-1080 cells. Among the HMC fractions, A2R is a strong candidate for anti-tumor targeting since its fraction exhibited better inhibition than the R and 2R fractions. Furthermore, the volume of the A2R fraction was approximately five times greater than that of the others, and included four proteins (molecular mass 9, 13, 22 and 40 kDa) detected by SDS-PAGE. Three of these (13, 22 and 40 kDa) were confirmed to be glycosylated with the Periodic Acid-Schiff Stain kit. These results suggest that A2R may possess anti-tumor activity and, in particular, that the protein components of A2R may act to selectively inhibit the growth of HT-1080 cells.

  20. A Compendium of Preparation and Application of Stem Cells in Parkinson's Disease: Current Status and Future Prospects.

    Science.gov (United States)

    Shen, Yan; Huang, Jinsha; Liu, Ling; Xu, Xiaoyun; Han, Chao; Zhang, Guoxin; Jiang, Haiyang; Li, Jie; Lin, Zhicheng; Xiong, Nian; Wang, Tao

    2016-01-01

    Parkinson's Disease (PD) is a progressively neurodegenerative disorder, implicitly characterized by a stepwise loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc) and explicitly marked by bradykinesia, rigidity, resting tremor and postural instability. Currently, therapeutic approaches available are mainly palliative strategies, including L-3,4-dihydroxy-phenylalanine (L-DOPA) replacement therapy, DA receptor agonist and deep brain stimulation (DBS) procedures. As the disease proceeds, however, the pharmacotherapeutic efficacy is inevitably worn off, worse still, implicated by side effects of motor response oscillations as well as L-DOPA induced dyskinesia (LID). Therefore, the frustrating status above has propeled the shift to cell replacement therapy (CRT), a promising restorative therapy intending to secure a long-lasting relief of patients' symptoms. By far, stem cell lines of multifarious origins have been established, which can be further categorized into embryonic stem cells (ESCs), neural stem cells (NSCs), induced neural stem cells (iNSCs), mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs). In this review, we intend to present a compendium of preparation and application of multifarious stem cells, especially in relation to PD research and therapy. In addition, the current status, potential challenges and future prospects for practical CRT in PD patients will be elaborated as well.

  1. Vismodegib, itraconazole and sonidegib as hedgehog pathway inhibitors and their relative competencies in the treatment of basal cell carcinomas.

    Science.gov (United States)

    Wahid, Mohd; Jawed, Arshad; Mandal, Raju K; Dar, Sajad A; Khan, Saif; Akhter, Naseem; Haque, Shafiul

    2016-02-01

    The advent of more sophisticated studies published has clarified the understating of the root cause of various skin cancers or basal cell carcinomas (BCCs). The remarkable role is played by the comprehensive work done on unraveling the mechanism controlling the function of hedgehog (Hh) pathway. The defective Hh pathway has been found as the major cause for BCCs as activated Hh signaling within primary cilia plays a key role in the pathogenesis of BCCs. The BCC accounts for up to 40% of all cancers in the US, with growing incidences in other countries as well. Thus, it is considered to be utmost important by the researchers all over the world developing drugs for the treatment of skin cancers targeting Hh pathway. Fewer drugs like vismodegib, itraconazole and sonidegib have shown promising results inhibiting the awry function of Hh pathway resulting in treatment of different forms of skin cancers. These drugs have shown positive results but failed to prove their potential as expected. Vismodegib and sonidegib are better but fail in case of resistant tumors. This review article describes the mechanism of actions of these Hh pathway inhibitors and provides the rationale for their effectiveness/non-effectiveness for the treatment of metastatic or locally advanced BCC.

  2. Gentamicin Blocks the ACh-Induced BK Current in Guinea Pig Type II Vestibular Hair Cells by Competing with Ca2+ at the l-Type Calcium Channel

    Directory of Open Access Journals (Sweden)

    Hong Yu

    2014-04-01

    Full Text Available Type II vestibular hair cells (VHCs II contain big-conductance Ca2+-dependent K+ channels (BK and L-type calcium channels. Our previous studies in guinea pig VHCs II indicated that acetylcholine (ACh evoked the BK current by triggering the influx of Ca2+ ions through l-type Ca2+ channels, which was mediated by M2 muscarinic ACh receptor (mAChRs. Aminoglycoside antibiotics, such as gentamicin (GM, are known to have vestibulotoxicity, including damaging effects on the efferent nerve endings on VHCs II. This study used the whole-cell patch clamp technique to determine whether GM affects the vestibular efferent system at postsynaptic M2-mAChRs or the membrane ion channels. We found that GM could block the ACh-induced BK current and that inhibition was reversible, voltage-independent, and dose-dependent with an IC50 value of 36.3 ± 7.8 µM. Increasing the ACh concentration had little influence on GM blocking effect, but increasing the extracellular Ca2+ concentration ([Ca2+]o could antagonize it. Moreover, 50 µM GM potently blocked Ca2+ currents activated by (--Bay-K8644, but did not block BK currents induced by NS1619. These observations indicate that GM most likely blocks the M2 mAChR-mediated response by competing with Ca2+ at the l-type calcium channel. These results provide insights into the vestibulotoxicity of aminoglycoside antibiotics on mammalian VHCs II.

  3. Comparison of commercial RNA extraction kits for preparation of DNA-free total RNA from Salmonella cells

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    Gonzalez-Escalona Narjol

    2010-07-01

    Full Text Available Abstract Background The isolation of DNA-free RNA is a crucial step in the reverse transcription PCR (RT-PCR. Every RNA extraction procedure results in RNA samples contaminated with genomic DNA, which can cause false-positive outcomes in highly sensitive applications, including a recently developed quantitative real-time PCR (RT-qPCR assay that targets invA mRNA for the detection of live Salmonella cells. The assay of this specific mRNA can be used to indicate the presence of live, as opposed to dead, cells of Salmonella enterica in a food matrix. Findings We evaluated the ability of five RNA extraction kits to produce RNA preparations from exponentially growing Salmonella cells. The acceptability of the preparations for use in downstream applications such as RT-qPCR was judged in terms of the total amount of RNA recovered, the integrity of the RNA molecules, and minimal content of DNA. The five kits produced RNA preparations that differed markedly in yield, integrity of the Salmonella RNA and the amount of contaminant DNA. The greatest RNA recovery was achieved with the MasterPure kit; however, the preparation contained high levels of genomic DNA. The UltraClean extraction kit gave a low level of RNA recovery with a poor level of integrity. The RNeasy Mini, RiboPure and PureLink extraction kits produced high-quality, DNA-free RNA suitable for Salmonella detection by RT-qPCR. Conclusions We showed that the RNeasy Mini and PureLink RNA extraction kits were the most suitable for the detection of Salmonella invA mRNA by RT-qPCR. The use of these two kits will greatly reduce the frequency of false-positive results and might allow fast RT-qPCR determination of invA mRNA produced by viable Salmonella in food samples.

  4. The utility of pap cell block preparations with liqui-PREP™ cell pellets to clarify the cytological diagnosis of atypical squamous cells of undetermined significance and atypical glandular cells.

    Science.gov (United States)

    George, Nicholas B; Baldassari, Jashua Haddad; Pérez Taveras, Digno A; José Fernández, María; Concepción Robledo, María

    2017-06-01

    Atypical squamous cells of undetermined significance (ASCUS) and atypical glandular cells (AGC) reflect cellular abnormalities insufficient for clear diagnosis. We used cell pellets obtained from liquid-based cytology (LBC) to prepare cell blocks (CB) and clarify the initial diagnosis of ASCUS and AGC. A total of 393 CBs with initial diagnosis of ASCUS or AGC were processed. Of those, 305 of the ASCUS and 20 of the AGC had adequate specimens. We compared results of CBs prepared from ASCUS and AGC to determine which had higher frequencies of higher-grade lesions. A majority of specimens (83%) were adequate for evaluation. Compared with the initial diagnosis, 14% (42/305) of ASCUS were diagnosed with low-grade squamous intraepithelial lesion (LSIL) in CB, while 10% (2/20) of AGC were diagnosed with LSIL or adenocarcinoma. No statistical relationship between the initial diagnosis of ASCUS and AGC and results of higher-grade lesions in CB is evident as determined by p value greater than 0.05 (p = 0.228). CBs prepared from Liqui-PREP cell pellets are, in most cases, assessable and can be useful as an adjunctive test to help clarify the initial diagnosis of ASCUS and AGC. Diagn. Cytopathol. 2017;45:520-525. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  5. New engineering: from knowledge to competences

    Science.gov (United States)

    Cartagena, M. C.; Tarquis, A. M.; Arce, A.

    2009-04-01

    One of the main innovations of Bologna system has been to link learning outcomes, ECTS workload based credits and competences. Competences represent a dynamic combination of knowledge, understanding, skills and abilities. Competences can be distinguished in subject specific and generic ones (instrumental, interpersonal and systemic competences). Actually in Spain Engineering degrees are changing to the new University educational system and should aim to satisfy the real needs of European society. This change has been long and complex, particularly. on the issue that have influenced curricular change Consultation with "actors" and "stakeholders", the definition of academic and professionals profiles and the translation of these into desired learning outcomes. Generic competences or transferable skills are relevant for preparing students well for their future role in society in terms of employability and citizenship. The criteria used by the companies to select their engineers are based in a good background and capacity to adapt and to acquire new knowledge, better than specific education, even postgraduate. It was interesting to note the great importance of generic competences However, Spanish government has regulated conditions of core curriculum need for to guarantee the acquisition of the competences needs to exercise the correspondent professional activities. The new degrees should comply with the core curriculum if the graduates want maintain the legal attributions guaranteed actually by the Spanish Professional Associations. After these degrees, students can access to professional master with actually horizontal attributions of regulated professions.

  6. Inhibition of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus

    Directory of Open Access Journals (Sweden)

    Chang HB

    2015-08-01

    Full Text Available Hong-Bin Chang,1 Bing-Huei Chen1,21Department of Food Science, 2Graduate Institute of Medicine, Fu Jen Catholic University, Taipei, TaiwanAbstract: The objectives of this study were to explore the inhibition mechanism of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus. In addition, human bronchus epithelial cell line BEAS-2B (normal cell was selected for comparison. A high-performance liquid chromatography (HPLC method was developed to separate and quantify the various curcuminoids in C. longa extract, including curcumin (1,714.5 µg/mL, demethoxycurcumin (1,147.4 µg/mL, and bisdemethoxycurcumin (190.2 µg/mL. A high-stability nanoemulsion composed of Tween 80, water, and curcuminoid extract was prepared, with mean particle size being 12.6 nm. The cell cycle was retarded at G2/M for both the curcuminoid extract and nanoemulsion treatments; however, the inhibition pathway may be different. H460 cells were more susceptible to apoptosis than A549 cells for both curcuminoid extract and nanoemulsion treatments. Growth of BEAS-2B remained unaffected for both the curcuminoid extract and nanoemulsion treatments, with a concentration range from 1 to 4 µg/mL. Also, the activities of caspase-3, caspase-8, and caspase-9 followed a dose-dependent increase for both A549 and H460 cells for both the treatments, accompanied by a dose-dependent increase in cytochrome C expression and a dose-dependent decrease in CDK1 expression. Interestingly, a dose-dependent increase in cyclin B expression was shown for A549 cells for both the treatments, while a reversed trend was found for H460 cells. Both mitochondria and death receptor pathways may be responsible for apoptosis of both A549 and H460 cells.Keywords: curcuminoid extract, curcuminoid nanoemulsion, Curcuma longa Linnaeus, lung cancer cell, cell cycle, apoptosis mechanism

  7. Preparation of single cells from aggregated Taxus suspension cultures for population analysis.

    Science.gov (United States)

    Naill, Michael C; Roberts, Susan C

    2004-06-30

    A method for the isolation of single plant cells from Taxus suspension cultures has been developed for the analysis of single cells via rapid throughput techniques such as flow cytometry. Several cell wall specific enzymes, such as pectinase, pectolyase Y-23, macerozyme, Driselase(R), and cellulase were tested for efficacy in producing single cell suspensions. The method was optimized for single cell yield, viability, time, and representivity of aggregated cell cultures. The best combination for single cell isolation was found to be 0.5% (w/v) pectolyase Y-23 and 0.04% (w/v) cellulase. High viability (>95%) and high yields of single cell aggregates (>90%) were obtained following 4 hours of digestion for four separate Taxus cell lines. In addition, methyl jasmonate elicitation (200 microM) was found to have no effect on three of the four tested Taxus lines. Isolated single cells were statistically similar to untreated cell cultures for peroxidase activity (model cell wall protein) and paclitaxel content (secondary metabolite produced in Taxus cell cultures). In comparison, protoplasts showed marked changes in both peroxidase activity and paclitaxel content as compared to untreated cultures. The use of flow cytometry was demonstrated with isolated cells that were found to have > 99% viability upon staining with fluorescein diacetate. The development of a method for the isolation of single plant cells will allow the study of population dynamics and culture variability on a single cell level for the development of population models of plant cell cultures and secondary metabolism. Copyright 2004 Wiley Periodicals, Inc.

  8. Preparation and properties of thin epoxy/compressed expanded graphite composite bipolar plates for proton exchange membrane fuel cells

    Science.gov (United States)

    Du, Chao; Ming, Pingwen; Hou, Ming; Fu, Jie; Shen, Qiang; Liang, Dong; Fu, Yunfeng; Luo, Xiaokuan; Shao, Zhigang; Yi, Baolian

    Although the composite bipolar plates prepared by the method of the vacuum resin impregnation in compressed expanded graphite (CEG) sheets have been applied in the KW-class stacks, there have been few investigations of the preparation and properties of them so far. In this research, the influences of the microstructure on the physical properties of the thin epoxy/CEG composites (the thickness is 1 mm) are investigated for the first time and the optimum preparation conditions are obtained. Results demonstrated that the mechanical property and the impermeability of the composites increases evidently with the resin content changing from 4% to 30%, while the electrical properties keep nearly constant. It can be attributed to the continuous expanded graphite (EG) conductive network of the raw CEG sheet. The epoxy (30 wt.%)/CEG composite is shown to be the optimum composite, displaying in-plane conductivity of 119.8 S cm -1, through-plane resistance of 17.13 mΩ cm 2, density of 1.95 g cm -3, gas permeability of 1.94 × 10 -6 cm 3 cm -2 s -1 and flexural strength of 45.8 MPa. The alcohol scrubbing is the optimum method of surface post-processing. The performance of a single cell with the optimum composite bipolar plates is tested and demonstrated to be outstanding. Above all, the composite prepared by resin vacuum impregnation in the CEG sheet is a promising candidate for bipolar plate materials in PEMFCs.

  9. Preparative isoelectric focusing and Joule effect: a purification cell that contains a heat exchanger.

    Science.gov (United States)

    Araque, A; Jaugey, J; Javet, P

    1996-01-01

    Reproducibility in protein purification by preparative isoelectric focusing depends greatly on temperature control during the separation process. A preparative apparatus is described, including a heat exchanger between compartments with isoelectric membranes. The selectivity of the isoelectric membranes was optimized as a function of isoelectric points of the separated proteins. At 2500 V and 60 W, 0.3 g of horse heart myoglobin from 0.2 g of whale skeletal muscle myoglobin could be separated in 1 h. At a total load of 2 g protein, 97% of bovine hemoglobin (2% initial concentration) was purified from bovine serum albumin (0.15%).

  10. Grief Counseling: An Investigation of Counselors' Training, Experience, and Competencies

    Science.gov (United States)

    Ober, Anne M.; Granello, Darcy Haag; Wheaton, Joe E.

    2012-01-01

    Grief is prevalent in counseling, but little is known about the current status of counselors' preparation and competencies to provide effective care. This exploratory study surveyed counselors (N = 369) on grief training, personal and professional experiences with grief, and grief counseling competence. Multiple regression analyses found training…

  11. Deliberations on the Development of an Intercultural Competence Curriculum

    Science.gov (United States)

    Punteney, Katherine

    2016-01-01

    Committed to developing an institution-wide intercultural competence curriculum for master's-level students preparing for international careers, a team of nine professors from across disciplines deliberated for a year on their fundamental understandings of intercultural competence and what it would mean to facilitate the development of that…

  12. Experimental cell transplantation therapy in rat myocardial infarction model including nude rat preparation.

    Science.gov (United States)

    Dai, Wangde; Kloner, Robert A

    2010-01-01

    As a novel potential therapeutic strategy for cardiac disease, cell transplantation therapy has been extensively investigated in experimental studies and clinical trials. Although encouraging results have been demonstrated, a number of critical questions still remain to be answered. For example, what kind of stem cell and how many cells should be used; what is the best time for cell transplantation after acute myocardial infarction; which delivery approach is better, intravenous injection or direct intramyocardial injection? Transplantation of cells derived from human tissues into experimental animals may elicit an immune rejection. Immunodeficient nude rats provide a useful myocardial infarction model for cell transplantation therapy studies. We introduce our detailed methods of direct intramyocardial injection of immature heart cells and stem cells into the myocardial infarction region of rats and nude rats. Careful maintenance under aseptic conditions and proper surgical technique are essential to improve the survival of immunodeficient rats after surgery.

  13. The use of gamma-irradiation and ultraviolet-irradiation in the preparation of human melanoma cells for use in autologous whole-cell vaccines

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    Denlinger Chadrick E

    2008-12-01

    Full Text Available Abstract Background Human cancer vaccines incorporating autologous tumor cells carry a risk of implantation and subsequent metastasis of viable tumor cells into the patient who is being treated. Despite the fact that the melanoma cell preparations used in a recent vaccine trial (Mel37 were gamma-irradiated (200 Gy, approximately 25% of the preparations failed quality control release criteria which required that the irradiated cells incorporate 3H-thymidine at no more than 5% the level seen in the non-irradiated cells. We have, therefore, investigated ultraviolet (UV-irradiation as a possible adjunct to, or replacement for gamma-irradiation. Methods Melanoma cells were gamma- and/or UV-irradiated. 3H-thymidine uptake was used to assess proliferation of the treated and untreated cells. Caspase-3 activity and DNA fragmentation were measured as indicators of apoptosis. Immunohistochemistry and Western blot analysis was used to assess antigen expression. Results UV-irradiation, either alone or in combination with gamma-irradiation, proved to be extremely effective in controlling the proliferation of melanoma cells. In contrast to gamma-irradiation, UV-irradiation was also capable of inducing significant levels of apoptosis. UV-irradiation, but not gamma-irradiation, was associated with the loss of tyrosinase expression. Neither form of radiation affected the expression of gp100, MART-1/MelanA, or S100. Conclusion These results indicate that UV-irradiation may increase the safety of autologous melanoma vaccines, although it may do so at the expense of altering the antigenic profile of the irradiated tumor cells.

  14. Learning Analytics for 21st Century Competencies

    Science.gov (United States)

    Buckingham Shum, Simon; Crick, Ruth Deakin

    2016-01-01

    Many educational institutions are shifting their teaching and learning towards equipping students with knowledge, skills, and dispositions that prepare them for lifelong learning, in a complex and uncertain world. These have been termed "21st century competencies." Learning analytics (LA) approaches in general offer different kinds of…

  15. Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins: Focus on sample preparation and derivatization conditions.

    Science.gov (United States)

    Weber, Daniela; Davies, Michael J; Grune, Tilman

    2015-08-01

    Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples.

  16. The preparation technique optimization of epoxy/compressed expanded graphite composite bipolar plates for proton exchange membrane fuel cells

    Science.gov (United States)

    Du, Chao; Ming, Pingwen; Hou, Ming; Fu, Jie; Fu, Yunfeng; Luo, Xiaokuan; Shen, Qiang; Shao, Zhigang; Yi, Baolian

    Vacuum resin impregnation method has been used to prepare polymer/compressed expanded graphite (CEG) composite bipolar plates for proton exchange membrane fuel cells (PEMFCs). In this research, three different preparation techniques of the epoxy/CEG composite bipolar plate (Compression-Impregnation method, Impregnation-Compression method and Compression-Impregnation-Compression method) are optimized by the physical properties of the composite bipolar plates. The optimum conditions and the advantages/disadvantages of the different techniques are discussed respectively. Although having different characteristics, bipolar plates obtained by these three techniques can all meet the demands of PEMFC bipolar plates as long as the optimum conditions are selected. The Compression-Impregnation-Compression method is shown to be the optimum method because of the outstanding properties of the bipolar plates. Besides, the cell assembled with these optimum composite bipolar plates shows excellent stability after 200 h durability testing. Therefore the composite prepared by vacuum resin impregnation method is a promising candidate for bipolar plate materials in PEMFCs.

  17. The preparation of HL-60 cells vaccine expressing BCG heat shock protein 70 and detection of its immunogenicity in vitro.

    Science.gov (United States)

    Li, Xiao-Ling; Zhao, Yan-Xia; Sun, Li-Rong; Yang, Jing; Xu, Hui-Juan

    2012-10-01

    Gene-modified cell vaccines are the best way to achieve the immunotherapy for all types of acute leukemia. In this study, the recombinant eukaryotic expression vector (pDisplay-HSP70) of heat shock protein 70 (HSP70) of Bacille Calmette-Guérin (BCG) was constructed by amplifying the whole BCG HSP70 gene using polymerase chain reaction (PCR) and sub-cloning into the polyclone endonuclease sites in pDisplay. Then the HL-60 cell vaccine expressing the protein onto the cell surface was prepared by lipofectamine transfection and its anti-tumor effect and mechanism were further studied. Results showed that the fragment of BCG HSP70 was consistent with Mycobacterium tuberculosis HSP70 gene published in GeneBank. DNA sequencing showed that the recombinant vector was correctly constructed and named pDisplay-HSP70. After BCG HSP70 gene transfection, the yellow-green fluorescence on the HL-60 cells surface was observed under a fluorescence microscope. The immunogenicity of HSP70-transfected HL-60 cells exhibited upregulated proliferation of lymphocytes, increased cytokine secretion (IFN-γ) and enhanced killing activity. These results suggested that gene transfection of BCG HSP70 could significantly enhance the immunogenicity of HL-60 cells. It may be used as a suitable candidate gene-modified cell vaccine for cancer immunotherapy.

  18. [Gelatin/alginate hydrogel scaffolds prepared by 3D bioprinting promotes cell adhesion and proliferation of human dental pulp cells in vitro].

    Science.gov (United States)

    Yu, Hai-Yue; Ma, Dan-Dan; Wu, Bu-Ling

    2017-05-20

    To evaluate the cytotoxicity of gelatin/alginate hydrogel scaffolds prepared by 3D bioprinting in human dental pulp cells (HDPCs) and compare the cell adhesion and proliferation of the cells seeded in the biomaterial using two different methods. HDPCs isolated by tissue block culture and enzyme digestion were cultured and passaged. Gelatin/alginate hydrogel scaffolds were printed using a bioplotter, and the cytotoxicity of the aqueous extracts of the scaffold material was tested in the third passage of HDPCs using cell counting kit-8. Scanning electron microscopy and trypan blue were used to assess the adhesion and proliferation of the cells seeded in the scaffold material at a low or high concentration. The aqueous extract of the scaffolds at different concentrations showed no obvious cytotoxicity and promoted the proliferation of HDPCs. The scaffolds had a good biocompatibility and HDPCs seeded in the scaffold showed good cell growth. Cell seeding at a high concentration in the scaffold better promoted the adhesion of HDPCs and resulted in a greater cell number on the scaffold surface compared with low-concentration cell seeding after a 5-day culture (Padhesion to the scaffold material.

  19. Preparation and characterization of nano-hydroxyapatite nanomaterials for liver cancer cell treatment.

    Science.gov (United States)

    Ezhaveni, Sathiyamoorthy; Yuvakkumar, Rathinam; Rajkumar, Mani; Sundaram, Nachiappan Meenakshi; Rajendran, Venkatachalam

    2013-03-01

    Nano-hydroxyapatite was synthesized by means of the hydrothermal treatment. The effects of nano-hydroxyapatite material on the behaviour of G2 liver cancer cells were explored. About 50% of cell viability was lost in nHAp material treated cells at 200 degrees C @ 5 h, followed by -30% in nHAp treated cells at 100 degrees C @ 5 h. Compared with control, nHAp material treated cells at 200 degrees C @ 5 h showed 60% and nHAp material treated cells at 100 degrees C @ 5 h showed 15% morphological change. Moreover, 50% of cell death was observed at 24 h incubation with nHAp material treated at 200 degrees C @ 5 h cells and 56% cell death at 48 h incubation and hence alters and disturbs the growth of cancer cells. In contrast, the nHAp material treated at 100 degrees C @ 5 h protects the cells and could be used for liver cancer cell treatment.

  20. Cultural Competence Among Italian Nurses: A Multicentric Survey.

    Science.gov (United States)

    Cicolini, Giancarlo; Della Pelle, Carlo; Comparcini, Dania; Tomietto, Marco; Cerratti, Francesca; Schim, Stephanie M; Di Giovanni, Pamela; Simonetti, Valentina

    2015-11-01

    To assess Italian nurses' cultural competence, as they are increasingly called upon to care for people of foreign origins. A cross-sectional, multicentric study. From September 2013 to May 2014, a survey was carried out among Italian nurses. Cultural competence was assessed by the Cultural Competence Assessment tool, translated and adapted to the Italian context. Nurses who completed the survey numbered 1,432; 70.6% were female; 42.6% ranged in age from 41 to 50 years; and 50.0% were bachelor's prepared. More than 50% had participated in some kind of cultural diversity training. Overall, cultural competence was moderate, showing a moderately high level of cultural awareness and sensitivity (mean = 5.41; SD = 0.66) and a moderate level of culturally competent behaviors (mean = 4.33; SD = 1.10). Although Italian nurses' cultural competence was acceptable, given the growing diversity of the patient population, nurses should be better prepared to face the changing health requests. Providing culturally competent care has been associated with improved provider-client communication, higher satisfaction with care, and health status improvement, as full comprehension of health status, adherence to medications and lifestyle recommendations, and appropriate utilization of the health system. Healthcare providers need to be adequately trained to provide culturally competent care. This research provides, for the first time, a report on Italian nurses' levels of cultural competence, and strengthens the current literature underlining the need for continuous education to enhance cultural competence among nurses. © 2015 Sigma Theta Tau International.

  1. [Preparation of a novel AAV-ITR gene expression mini vector in Sf9 insect cells via baculovirus].

    Science.gov (United States)

    Li, Taiming; Pan, Junjie; Qi, Jing; Zhang, Chun

    2015-08-01

    AAV-ITR gene expression mini vector is a double-strand or single-strand DNA that only contains inverted terminal repeats of adeno-associated virus, cis-elements and gene of interest and does not contain any other foreign DNA sequences. We prepared Bac-ITR-EGFP and Bac-inrep. Spodoptera frugiperda cells were infected with Bac-ITR-EGFP (P3) and Bac-inrep (P3). Up to 100 μg of AAV-ITR-EGFP gene expression mini vectors were extracted from 2 x 10(7) cells of Sf9 72 h after infection. The gel electrophoresis analysis shows that most forms of AAV-ITR-EGFP gene expression mini vector were monomer and dimer. The mini vector expression efficacy was examined in vitro with HEK 293T cells. The EGFP expression was observed at 24 h after transfection, and the positive ratio reached 65% at 48 h after transfection.

  2. Preparation of Semiconducting Materials in the Laboratory, Part 3: The One-Penny Photovoltaic Cell

    Science.gov (United States)

    Ibanez, Jorge G.; Finck-Pastrana, Adolfo; Mugica-Barrera, Alejandra; Balderas-Hernandez, Patricia; Ibarguengoitia-Cervantes, Martha E.; Garcia-Pintor, Elizabeth; Hartasanchez-Frenk, Jose Miguel; Bonilla-Jaurez, Cesar E.; Maldonado-Cordero, Casandra; Struck-Garza, Adelwart; Suberbie-Rocha, Felipe

    2011-01-01

    Copper(I) oxide photoresponsive layers are prepared on copper surfaces (e.g., U.S. pre-1982 pennies) by simple thermal, chemical, and electrochemical procedures. An easily measurable photovoltage (up to 100 mV) is obtained in each case under visible light illumination. (Contains 2 figures.)

  3. Preparation of Semiconducting Materials in the Laboratory, Part 3: The One-Penny Photovoltaic Cell

    Science.gov (United States)

    Ibanez, Jorge G.; Finck-Pastrana, Adolfo; Mugica-Barrera, Alejandra; Balderas-Hernandez, Patricia; Ibarguengoitia-Cervantes, Martha E.; Garcia-Pintor, Elizabeth; Hartasanchez-Frenk, Jose Miguel; Bonilla-Jaurez, Cesar E.; Maldonado-Cordero, Casandra; Struck-Garza, Adelwart; Suberbie-Rocha, Felipe

    2011-01-01

    Copper(I) oxide photoresponsive layers are prepared on copper surfaces (e.g., U.S. pre-1982 pennies) by simple thermal, chemical, and electrochemical procedures. An easily measurable photovoltage (up to 100 mV) is obtained in each case under visible light illumination. (Contains 2 figures.)

  4. Preparation and characterization of compact TiO2 film used in Gr(a)tzel solar cells

    Institute of Scientific and Technical Information of China (English)

    LI Bin; WANG Liduo; ZHANG Deqiang; QIU Yong

    2004-01-01

    We report for the first time the preparation method of compact TiO2 film using titanium butoxide-petroleum ether solution, by means of dipping and sintering. Our estimation of the component of this thin film was confirmed by XPS test. Studies of AFM showed the influence of preparation condition upon surface morphology. The transmitted spectrum of this compact TiO2 film was also investigated, which presented an excellent transmittance of this film in the visible light region. We arrive at a conclusion, according to the I-V characteristics of solid-state Gratzel solar cells utilizing this kind of compact film, that this compact TiO2 film functions well in secluding the conducting substrate from the hole-conducting material.

  5. Fermented Papaya Preparation Restores Age-Related Reductions in Peripheral Blood Mononuclear Cell Cytolytic Activity in Tube-Fed Patients

    Science.gov (United States)

    Fujita, Yuhzo; Tsuno, Haruo; Nakayama, Jiro

    2017-01-01

    Tube-fed elderly patients are generally supplied with the same type of nutrition over long periods, resulting in an increased risk for micronutrient deficiencies. Dietary polyphenols promote immunity and have anti-inflammatory, anti-carcinogenic, and anti-oxidative properties. Carica papaya Linn. is rich in several polyphenols; however, these polyphenols are poorly absorbed from the digestive tract in their original polymerized form. Therefore, we determined the molecular components of a fermented Carica papaya Linn. preparation, as well as its effects on immunity and the composition of gut microbiota in tube-fed patients. Different doses of the fermented C. papaya L. preparation were administered to three groups of tube-fed patients for 30 days. Its effects on fecal microbiota composition and immunity were assessed by 16S rRNA gene sequencing and immune-marker analysis, respectively. The chemical composition of the fermented C. papaya L. preparation was analyzed by capillary electrophoresis- and liquid chromatography- time of flight mass spectrometry. The fermented C. papaya L. preparation restored peripheral blood mononuclear cell (PBMC) cytolytic activity; however, no other biomarkers of immunity were observed. Treatment with the preparation (9 g/day) significantly reduced the abundance of Firmicutes in the fecal microbiota. In particular, treatment reduced Clostridium scindens and Eggerthella lenta in most patients receiving 9 g/day. Chemical analysis identified low-molecular-weight phenolic acids as polyphenol metabolites; however, no polymerized, large-molecular-weight molecules were detected. Our study indicates that elderly patients who are tube-fed over the long-term have decreased PBMC cytolytic activity. In addition, low-molecular-weight polyphenol metabolites fermented from polymerized polyphenols restore PBMC cytolytic activity and modulate the composition of gut microbiota in tube-fed patients. PMID:28060858

  6. Preparation of cell-lines for conditional knockdown of gene expression and measurement of the knockdown effects on E4orf4-induced cell death.

    Science.gov (United States)

    Brestovitsky, Anna; Sharf, Rakefet; Kleinberger, Tamar

    2012-10-21

    Functional inactivation of gene expression in mammalian cells is crucial for the study of the contribution of a protein of interest to various pathways(1,2). However, conditional knockdown of gene expression is required in cases when constitutive knockdown is not tolerated by cells for a long period of time(3-5). Here we describe a protocol for preparation of cell lines allowing conditional knockdown of subunits of the ACF chromatin remodeling factor. These cell lines facilitate the determination of the contribution of ACF to induction of cell death by the adenovirus E4orf4 protein(6). Sequences encoding short hairpin RNAs for the Acf1 and SNF2h subunits of the ACF chromatin remodeling factor were cloned next to a doxycycline-inducible promoter in a plasmid also containing a gene for the neomycin resistance gene. Neomycin-resistant cell clones were selected in the presence of G418 and isolated. The resulting cell lines were induced by doxycycline treatment, and once Acf1 or SNF2h expression levels were reduced, the cells were transfected with a plasmid encoding E4orf4 or an empty vector. To confirm the specific effect of the shRNA constructs, Acf1 or SNF2h protein levels were restored to WT levels by cotransfection with a plasmid expressing Acf1 or SNF2h which were rendered resistant to the shRNA by introduction of silent mutations. The ability of E4orf4 to induce cell death in the various samples was determined by a DAPI assay, in which the frequency of appearance of nuclei with apoptotic morphologies in the transfected cell population was measured(7-9). The protocol described here can be utilized for determination of the functional contribution of various proteins to induction of cell death by their protein partners in cases when constitutive knockdown may be cell lethal.

  7. Corrigendum to "Preparation of Nafion-sulfonated clay nanocomposite membrane for direct menthol fuel cells via a film coating process" [J. Power Sources 165 (2007) 1-2

    Science.gov (United States)

    Kim, Tae Kyoung; Kang, Myeongsoon; Choi, Yeong Suk; Kim, Hae Kyung; Lee, Wonmok; Chang, Hyuk; Seung, Doyoung

    The author regrets that the above paper was printed with an error in the title. The correct title reads: "Preparation of Nafion-sulfonated clay nanocomposite membrane for direct methanol fuel cells via a film coating process".

  8. Competence and competency of high school teacher as the components of his innovative development

    Directory of Open Access Journals (Sweden)

    I. K. Yarmola

    2013-10-01

    Full Text Available Priority direction of state policy concerning the development of higher education inUkraineis to prepare qualified and competitive staff in the labor market. The high level of competence and scientific competence along with pedagogical craftsmanship and psychological culture of teachers is condition for improving the efficiency of the educational process in higher education. Aim of research was theoretical justification importance of competence and competency of high school teacher for the establishment his innovation potential. In the article the analysis of interpretation of the terms competence and competency in various scientific sources and scientific works of individual authors were presented. Moreover important are ability to creativity, solving of problem’s tasks, ingenuity, flexibility and criticality mind, intuition, originality and self-confidence; ability to formulate and solve non-standard tasks; ability to analyse, synthesis and combination, to share experiences, predictions, etc.. Emotionally-shaped quality represented by the following components as spirituality, the emotional upsurge in creative situations; associativity, imagination, sense of novelty, sensitivity to contradictions, empathy; insight, ability to see the familiar in an unfamiliar; overcome the stereotypes; risk appetite, desire for freedom. The general regularities of formation of research competence have been detected. In particular, the formation carried out through the development of competencies invariant character and increasing of specialized skills. It is proved that scientific and research competence is determined primarily by the system of teacher's professional education and self-education. The most important objective characteristics that reflect the competence of teachers in the field R & D is the total number of publications, number of scientific articles, monographs, and also won grants, competitions, etc. Moreover the level of competence in the

  9. Preparation, characterization and toxicological investigation of copper loaded chitosan nanoparticles in human embryonic kidney HEK-293 cells

    Energy Technology Data Exchange (ETDEWEB)

    Arora, Divya [Academy of Scientific and Innovative Research (AcSIR), CSIR-Indian Institute of Integrative Medicine, Jammu (India); Formulation and Drug Delivery Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Dhanwal, Vandna [Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Nayak, Debasis [Academy of Scientific and Innovative Research (AcSIR), CSIR-Indian Institute of Integrative Medicine, Jammu (India); Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Saneja, Ankit [Academy of Scientific and Innovative Research (AcSIR), CSIR-Indian Institute of Integrative Medicine, Jammu (India); Formulation and Drug Delivery Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Amin, Hina [Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Rasool, Reyaz ur [Academy of Scientific and Innovative Research (AcSIR), CSIR-Indian Institute of Integrative Medicine, Jammu (India); Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Gupta, Prem Narayan [Academy of Scientific and Innovative Research (AcSIR), CSIR-Indian Institute of Integrative Medicine, Jammu (India); Formulation and Drug Delivery Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Goswami, Anindya, E-mail: agoswami@iiim.ac.in [Academy of Scientific and Innovative Research (AcSIR), CSIR-Indian Institute of Integrative Medicine, Jammu (India); Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India)

    2016-04-01

    Metallic nanoparticles often attribute severe adverse effects to the various organs or tissues at the molecular level despite of their applications in medical, laboratory and industrial sectors. The present study highlights the preparation of copper adsorbed chitosan nanoparticles (CuCSNPs), its characterization and validation of cytotoxicity in human embryonic kidney HEK-293 cells. Particle size of the CuCSNPs was determined by using Zetasizer and the copper loading was quantified with the help of ICP/MS. Further characterization of CuCSNPs was carried out by FT-IR analysis to determine the formation of nanoparticles and SEM was conducted for the morphological analysis of the CuCSNPs. The CuCSNPs exhibited pronounced cytotoxic effects towards HEK-293 cells as analyzed by MTT assay. Moreover, the CuCSNPs inhibited the colony formation and induced nuclear damage at the dose of 100 μg/mL, much more effectively than the in built control copper sulfate (CuSO{sub 4}). At the molecular level, the CuCSNPs were found to be triggering reactive oxygen species (ROS), activating effector caspases and subsequent PARP cleavage to induce cell death in HEK-293 cells. - Highlights: • Subtoxic levels of CuCSNPs induce apoptosis in HEK-293 cells. • CuCSNPs mediate toxicity via nuclear cleavage and ROS generation. • CuCSNPs favor caspase activation and PARP cleavage to induce cell death.

  10. Antioxidant activity and growth inhibition of human colon cancer cells by crude and purified fucoidan preparations extracted from Sargassum cristaefolium

    Directory of Open Access Journals (Sweden)

    Cheng-Yuan Wang

    2015-12-01

    Full Text Available Fucose-containing sulfated polysaccharides, also termed “fucoidans”, which are known to possess antioxidant, anticoagulant, anticancer, antiviral, and immunomodulating properties, are normally isolated from brown algae via various extraction techniques. In the present study, two methods (SC1 and SC2 for isolation of fucoidan from Sargassum cristaefolium were compared, with regard to the extraction yields, antioxidant activity, and inhibition of growth of human colon cancer cells exhibited by the respective extracts. SC1 and SC2 differ in the number of extraction steps and concentration of ethanol used, as well as the obtained sulfated polysaccharide extracts, namely, crude fucoidan preparation (CFP and purified fucoidan preparation (PFP, respectively. Thin layer chromatography, Fourier transform infrared analysis, and measurements of fucose and sulfate contents revealed that the extracts were fucoidan. There was a higher extraction yield for CFP, which contained less fucose and sulfate but more uronic acid, and had weaker antioxidant activity and inhibition of growth in human colon cancer cells. In contrast, there was a lower extraction yield for PFP, which contained more fucose and sulfate but less uronic acid, and had stronger antioxidant activity and inhibition of growth in human colon cancer cells. Thus, since the difference in bioactive activities between CFP and PFP was not remarkable, the high extraction yield of SC1 might be favored as a method in industrial usage for extracting fucoidan.

  11. Regulation of Varicella-Zoster Virus-Induced Cell-to-Cell Fusion by the Endocytosis-Competent Glycoproteins gH and gE

    OpenAIRE

    Pasieka, Tracy Jo; Maresova, Lucie; Shiraki, Kimiyasu; Grose, Charles

    2004-01-01

    The gH glycoprotein of varicella-zoster virus (VZV) is a major fusogen. The realigned short cytoplasmic tail of gH (18 amino acids) harbors a functional endocytosis motif (YNKI) that mediates internalization in both VZV-infected and transfected cells (T. J. Pasieka, L. Maresova, and C. Grose, J. Virol. 77: 4194-4202, 2003). During subsequent confocal microscopy studies of endocytosis-deficient gH mutants, we observed that cells transfected with the gH tail mutants exhibited marked fusion. The...

  12. Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.

    Directory of Open Access Journals (Sweden)

    Maria Jesús Cánepa

    Full Text Available Reproductive biotechnologies such as in vitro fertilization (IVF and somatic cell nuclear transfer (SCNT enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70, endoplasmic reticulum (ER stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5 and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3 in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART.

  13. Series-Interconnected Plastic Dye-Sensitized Solar Cells Prepared by Low- Temperature Binder-Free Titania Paste

    Directory of Open Access Journals (Sweden)

    Erlyta Septa Rosa

    2014-10-01

    Full Text Available The aim of this research is to study dye-sensitized solar cells (DSSC. This was implemented on a flexible polyethylene terephthalate (PET substrate using a mixture of transparent and scattered mesoporous anatase-titania as the electron transport layer for the photoelectrode. This mixture of anatase titania performed a dual function of light scattering and efficient dye absorption. In this study, a porous nano-TiO2 film was prepared on indium tin oxide (ITO coated polyethylene terephthalate (PET by using a binder-free titania paste; on it, a DSSC was fabricated. The paste which contained a mixture of TiO2 nanoparticles, acid chloride, and ethanol was printed on two patterns of 1x6 cm2 active areas followed by sintered at 120 ºC to form TiO2 films. A commercial dye, N719, was adsorbed on the surface of TiO2 films and assembled to two platinized conductive plastic patterns to form a counter electrode and thus a sandwich-type dye cell. Finally, a solution of KI/I2 electrolytes was injected into the cell in which a couple of sandwich-type dye cells with an active area of 6 cm2 for each cell were series interconnected with a z-type interconnection between the photoelectrode of one cell and the counter electrode of another cell. The cell performance was characterized by employing simulated solar light at an intensity of 50 mW/cm2. The results showed interconnected cells generating a short-circuit photocurrent density of 2.34 mA/cm2, an open-circuit voltage of 1.10 volt, and overall 0.172% power conversion efficiency.

  14. Complex heterogeneous tissue constructs containing multiple cell types prepared by inkjet printing technology.

    Science.gov (United States)

    Xu, Tao; Zhao, Weixin; Zhu, Jian-Ming; Albanna, Mohammad Z; Yoo, James J; Atala, Anthony

    2013-01-01

    This study was designed to develop a versatile method for fabricating complex and heterogeneous three-dimensional (3D) tissue constructs using simultaneous ink-jetting of multiple cell types. Human amniotic fluid-derived stem cells (hAFSCs), canine smooth muscle cells (dSMCs), and bovine aortic endothelial cells (bECs), were separately mixed with ionic cross-linker calcium chloride (CaCl(2)), loaded into separate ink cartridges and printed using a modified thermal inkjet printer. The three cell types were delivered layer-by-layer to pre-determined locations in a sodium alginate-collagen composite located in a chamber under the printer. The reaction between CaCl(2) and sodium alginate resulted in a rapid formation of a solid composite gel and the printed cells were anchored in designated areas within the gel. The printing process was repeated for several cycles leading to a complex 3D multi-cell hybrid construct. The biological functions of the 3D printed constructs were evaluated in vitro and in vivo. Each of the printed cell types maintained their viability and normal proliferation rates, phenotypic expression, and physiological functions within the heterogeneous constructs. The bioprinted constructs were able to survive and mature into functional tissues with adequate vascularization in vivo. These findings demonstrate the feasibility of fabricating complex heterogeneous tissue constructs containing multiple cell types using inkjet printing technology.

  15. In vitro transcription of a cloned vaccinia virus gene by a soluble extract prepared from vaccinia virus-infected HeLa cells.

    OpenAIRE

    Foglesong, P D

    1985-01-01

    Faithful transcription of a vaccinia virus gene was accomplished in vitro by using a soluble extract prepared from vaccinia virus-infected HeLa cells. Specific transcription of the cloned vaccinia virus gene was detected by using template DNA restricted within the transcribed region. The vaccinia virus gene was not transcribed by extracts prepared from uninfected HeLa cells even with supplementation by purified vaccinia virus RNA polymerase, nor was a clone of adenovirus 2 DNA bearing the maj...

  16. Preparation, characterization, and high performance of RuSe/C for direct methanol fuel cells

    Science.gov (United States)

    Serov, Alexey Alexandrovich; Min, Myoungki; Chai, Geunseok; Han, Sangil; Kang, Soonki; Kwak, Chan

    RuSe/C catalysts prepared by different methods have been tested for oxygen reduction and the results analyzed based on the active species and particle size distribution. Inorganic precursor methods exhibit higher catalytic performance than the carbonyl method. Selenious acid is an excellent inorganic precursor. X-ray photoelectron spectra indicate that selenium in a high oxidation state is more active than that with zero valence. The effect of operating conditions is analyzed for catalysts prepared by the inorganic precursor method. The optimum heat-treatment temperature for both active phase formation and particle size distribution is 300 °C. A performance of 62 mW cm -2 at 80 °C is obtained using 80 wt.% RuSe/C in the cathode.

  17. Preparation, Cell Compatibility and Degradability of Collagen-Modified Poly(lactic acid)

    OpenAIRE

    Miaomiao Cui; Leili Liu; Ning Guo; Ruixia Su; Feng Ma

    2015-01-01

    Poly(lactic acid) (PLA) was modified using collagen through a grafting method to improve its biocompatibility and degradability. The carboxylic group at the open end of PLA was transferred into the reactive acylchlorided group by a reaction with phosphorus pentachloride. Then, collagen-modified PLA (collagen-PLA) was prepared by the reaction between the reactive acylchlorided group and amino/hydroxyl groups on collagen. Subsequently, the structure of collagen-PLA was confirmed by Fourier t...

  18. Influencing Selectivity to Cancer Cells with Mixed Nanoparticles Prepared from Albumin-Polymer Conjugates and Block Copolymers.

    Science.gov (United States)

    Jiang, Yanyan; Wong, Sandy; Chen, Fan; Chang, Ted; Lu, Hongxu; Stenzel, Martina H

    2017-04-19

    Albumin-based nanoparticles are widely used to delivery anticancer drug because they promote the accumulation of drugs in tumor sites. Nanoparticles with surface immobilized albumin are widely described in literature, although mixed nanoparticles with systematically modified ratios between albumin and PEG-based material are less common. In this work, hybrid nanoparticles were prepared by coassembly of a PEG-based amphiphilic block copolymer together with a polymer-protein conjugate. Poly(oligo(ethylene glycol) methyl ether acrylate)-poly(ε-caprolactone) (POEGMEA-PCL) was prepared by a combination of ring-opening polymerization and reversible addition-fragmentation chain transfer (RAFT) polymerization, while the polymer-protein conjugate was obtained by reacting poly(ε-caprolactone) with bovine serum albumin (BSA-PCL). Co-assembly of both amphiphiles at different ratios, with and without curcumin as a drug, led to hybrid nanoparticles with various amount of albumin on the particle surface. The resulting hybrid nanoparticles were similar in size (100-120 nm), but increasing the amount of albumin on the surface led to a more-negative ζ potential. The cytotoxicity of the curcumin-loaded nanoparticles was examined on several cell lines. The curcumin-loaded nanoparticles with high amount of albumin led to high cytotoxicity against breast cancer cell lines (MDA-MB-231 and MCF-7), which coincided with high cellular uptake. However, the cytotoxicity of the curcumin-loaded nanoparticles against CHO cells and RAW264.7 cells was reduced, suggesting that albumin can facilitate selectivity toward cancer cells.

  19. Preparation, quality criteria, and properties of human blood platelet lysate supplements for ex vivo stem cell expansion.

    Science.gov (United States)

    Shih, Daniel Tzu-Bi; Burnouf, Thierry

    2015-01-25

    Most clinical applications of human multipotent mesenchymal stromal cells (MSCs) for cell therapy, tissue engineering, regenerative medicine, and treatment of immune and inflammatory diseases require a phase of isolation and ex vivo expansion allowing a clinically meaningful cell number to be reached. Conditions used for cell isolation and expansion should meet strict quality and safety requirements. This is particularly true for the growth medium used for MSC isolation and expansion. Basal growth media used for MSC expansion are supplemented with multiple nutrients and growth factors. Fetal bovine serum (FBS) has long been the gold standard medium supplement for laboratory-scale MSC culture. However, FBS has a poorly characterized composition and poses risk factors, as it may be a source of xenogenic antigens and zoonotic infections. FBS has therefore become undesirable as a growth medium supplement for isolating and expanding MSCs for human therapy protocols. In recent years, human blood materials, and most particularly lysates and releasates of platelet concentrates have emerged as efficient medium supplements for isolating and expanding MSCs from various origins. This review analyzes the advantages and limits of using human platelet materials as medium supplements for MSC isolation and expansion. We present the modes of production of allogeneic and autologous platelet concentrates, measures taken to ensure optimal pathogen safety profiles, and methods of preparing PLs for MSC expansion. We also discuss the supply of such blood preparations. Produced under optimal conditions of standardization and safety, human platelet materials can become the future 'gold standard' supplement for ex vivo production of MSCs for translational medicine and cell therapy applications.

  20. Preparation of luminescent CdTe quantum dots doped core-shell nanoparticles and their application in cell recognition

    Institute of Scientific and Technical Information of China (English)

    LI Zhaohui; WANG Kemin; TAN Weihong; LI Jun; FU Zhiying; WANG Yilin; LIU Jianbo; YANG Xiaohai

    2005-01-01

    Based on the reverse microemulsion technique, luminescent quantum dots doped core-shell nanoparticles have been prepared by employing silica as the shell and CdTe quantum dots as the core of the nanoparticles, which have an excellent solubility and dispersibility, especially amine and phosphonate groups have been modified on their surface synchronously. In comparison with CdTe quantum dots, these nanoparticles show superiority in chemical and photochemical stability. The quantum dots doped core-shell nanoparticles were successfully linked with lactobionic acid by amine group on it, which was used to recognize living liver cells.

  1. A pull-down method with a biotinylated bait protein prepared by cell-free translation using a puromycin linker.

    Science.gov (United States)

    Mochizuki, Yuki; Kohno, Fumiaki; Nishigaki, Koichi; Nemoto, Naoto

    2013-03-01

    In this paper, we demonstrate a novel pull-down method that dramatically reduces the cost and preparation time of a bait protein by cell-free translation with a puromycin linker. With the C-terminus of the bait protein linked to biotin through a puromycin molecule after the translation reaction and subsequent mRNA degradation by RNase, the prey protein was easily pulled down by streptavidin-coated magnetic beads in a test tube. Three fluorescent prey protein types were tested and confirmed by gel electrophoresis to be pulled down easily and rapidly, depending on their affinity. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. Colocalization and interaction between elongasome and divisome during a preparative cell division phase in Escherichia coli

    NARCIS (Netherlands)

    van der Ploeg, R.; Verheul, J.; Vischer, N.O.E.; Alexeeva, S.; Hoogendoorn, E.; Postma, M.; Banzhaf, M.; Vollmer, W.; den Blaauwen, T.

    2013-01-01

    The rod-shaped bacterium Escherichia coli grows by insertion of peptidoglycan into the lateral wall during cell elongation and synthesis of new poles during cell division. The monofunctional transpeptidases PBP2 and PBP3 are part of specialized protein complexes called elongasome and divisome,

  3. A solid oxide fuel cell with a gadolinia-doped ceria anode: Preparation and performance

    DEFF Research Database (Denmark)

    Marina, O.A.; Bagger, C.; Primdahl, S.

    1999-01-01

    /N-2 = 33/3/64. No carbon deposition was found on CG4 after cell operation at a steam-to-carbon ratio of 0.3 for 1000 h. Cells sustained several rapid thermal cycles in the temperature interval 200-1000 degrees C and a full redox cycle without degradation. (C) 1999 Elsevier Science B.V. All rights...

  4. Colocalization and interaction between elongasome and divisome during a preparative cell division phase in Escherichia coli

    NARCIS (Netherlands)

    Ploeg, van der R.; Verheul, J.; Vischer, N.O.E.; Alexeeva, S.V.; Hoogendoorn, E.; Postma, M.; Banzhaf, M.; Vollmer, W.; Blaauwen, den T.

    2013-01-01

    The rod-shaped bacterium Escherichia coli grows by insertion of peptidoglycan into the lateral wall during cell elongation and synthesis of new poles during cell division. The monofunctional transpeptidases PBP2 and PBP3 are part of specialized protein complexes called elongasome and divisome, respe

  5. Establishment of Trophectoderm Cell Lines from Buffalo (Bubalus bubalis Embryos of Different Sources and Examination of In Vitro Developmental Competence, Quality, Epigenetic Status and Gene Expression in Cloned Embryos Derived from Them.

    Directory of Open Access Journals (Sweden)

    Sushil Kumar Mohapatra

    Full Text Available Despite being successfully used to produce live offspring in many species, somatic cell nuclear transfer (NT has had a limited applicability due to very low (>1% live birth rate because of a high incidence of pregnancy failure, which is mainly due to placental dysfunction. Since this may be due to abnormalities in the trophectoderm (TE cell lineage, TE cells can be a model to understand the placental growth disorders seen after NT. We isolated and characterized buffalo TE cells from blastocysts produced by in vitro fertilization (TE-IVF and Hand-made cloning (TE-HMC, and compared their growth characteristics and gene expression, and developed a feeder-free culture system for their long-term culture. The TE-IVF cells were then used as donor cells to produce HMC embryos following which their developmental competence, quality, epigenetic status and gene expression were compared with those of HMC embryos produced using fetal or adult fibroblasts as donor cells. We found that although TE-HMC and TE-IVF cells have a similar capability to grow in culture, significant differences exist in gene expression levels between them and between IVF and HMC embryos from which they are derived, which may have a role in the placental abnormalities associated with NT pregnancies. Although TE cells can be used as donor cells for producing HMC blastocysts, their developmental competence and quality is lower than that of blastocysts produced from fetal or adult fibroblasts. The epigenetic status and expression level of many important genes is different in HMC blastocysts produced using TE cells or fetal or adult fibroblasts or those produced by IVF.

  6. A COMPARATIVE STUDY OF FLUID CYTOLOGY WITH SMEAR AND CELL BLOCK PREPARATION

    Directory of Open Access Journals (Sweden)

    Aruna Padmavathi

    2016-08-01

    Full Text Available BACKGROUND Exfoliative cytology has got a key role in the diagnosis of variable pathological conditions. This technique is noninvasive and performed in the practice of cytopathology. The cell block technique now takes an intermediate position between histological and cytological techniques. METHODS A prospective study was done from October 2009 to October 2011 in the Department of Pathology. RESULTS Among 140 fluids, 12 were clinically diagnosed as malignant effusions out of which 11 were reported as positive for malignant cells and 1 case was negative. 12 cases were reported as positive for malignant cells in which they were clinically not diagnosed. CONCLUSIONS In the present study, increased diagnostic utility of 10% is noted in cell block method. The cell block technique not only increased the positive results but also helped to demonstrate better architectural patterns which could be of great help in approaching the correct diagnosis of the primary site.

  7. Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions.

    Science.gov (United States)

    Broggi, Achille; Cigni, Clara; Zanoni, Ivan; Granucci, Francesca

    2016-04-20

    The skin is a barrier organ that interacts with the external environment. Being continuously exposed to potential microbial invasion, the dermis and epidermis home a variety of immune cells in both homeostatic and inflammatory conditions. Tools to obtain skin cell release for cytofluorimetric analyses are, therefore, very useful in order to study the complex network of immune cells residing in the skin and their response to microbial stimuli. Here, we describe an efficient methodology for the digestion of mouse skin to rapidly and efficiently obtain single-cell suspensions. This protocol allows maintenance of maximum cell viability without compromising surface antigen expression. We also describe how to take and digest skin samples from different anatomical locations, such as the ear, trunk, tail, and footpad. The obtained suspensions are then stained and analyzed by flow cytometry to discriminate between different leukocyte populations.

  8. Fabrication and Photovoltaic Characteristics of Coaxial Silicon Nanowire Solar Cells Prepared by Wet Chemical Etching

    Directory of Open Access Journals (Sweden)

    Chien-Wei Liu

    2012-01-01

    Full Text Available Nanostructured solar cells with coaxial p-n junction structures have strong potential to enhance the performances of the silicon-based solar cells. This study demonstrates a radial junction silicon nanowire (RJSNW solar cell that was fabricated simply and at low cost using wet chemical etching. Experimental results reveal that the reflectance of the silicon nanowires (SNWs declines as their length increases. The excellent light trapping was mainly associated with high aspect ratio of the SNW arrays. A conversion efficiency of ∼7.1% and an external quantum efficiency of ∼64.6% at 700 nm were demonstrated. Control of etching time and diffusion conditions holds great promise for the development of future RJSNW solar cells. Improving the electrode/RJSNW contact will promote the collection of carries in coaxial core-shell SNW array solar cells.

  9. Changes in pituitary growth hormone cells prepared from rats flown on Spacelab 3

    Science.gov (United States)

    Grindeland, R.; Hymer, W. C.; Farrington, M.; Fast, T.; Hayes, C.; Motter, K.; Patil, L.; Vasques, M.

    1987-01-01

    The effect of exposure to microgravity on pituitary gland was investigated by examining cells isolated from anterior pituitaries of rats flown on the 7-day Spacelab 3 mission and, subsequently, cultured for 6 days. Compared with ground controls, flight cells contained more intracellular growth hormone (GH); however, the flight cells released less GH over the 6-day culture period and after implantation into hypophysectomized rats than did the control cells. Compared with control rats, glands from large rats (400 g) contained more somatotrophs (44 percent compared with 37 percent in control rats); small rats (200 g) showed no difference. No major differences were found in the somatotroph ultrastructure (by TEM) or in the pattern of the immunoactive GH variants. However, high-performance liquid chromatography fractionation of culture media indicated that flight cells released much less of a biologically active high-molecular weight GH variant, suggesting that space flight may lead to secretory dysfunction.

  10. Nurse Leadership and Informatics Competencies: Shaping Transformation of Professional Practice.

    Science.gov (United States)

    Kennedy, Margaret Ann; Moen, Anne

    2017-01-01

    Nurse leaders must demonstrate capacities and develop specific informatics competencies in order to provide meaningful leadership and support ongoing transformation of the healthcare system. Concurrently, staff informatics competencies must be planned and fostered to support critical principles of transformation and patient safety in practice, advance evidence-informed practice, and enable nursing to flourish in complex digital environments across the healthcare continuum. In addition to nurse leader competencies, two key aspects of leadership and informatics competencies will be addressed in this chapter - namely, the transformation of health care and preparation of the nursing workforce.

  11. Ethanol Extract of Abnormal Savda Munziq, a Herbal Preparation of Traditional Uighur Medicine, Inhibits Caco-2 Cells Proliferation via Cell Cycle Arrest and Apoptosis

    Directory of Open Access Journals (Sweden)

    Abdiryim Yusup

    2012-01-01

    Full Text Available Aims. Study the effect of Abnormal Savda Munziq (ASMq ethanol extract on the proliferation, apoptosis, and correlative gene, expression in colon cancer cells (Caco-2 to elucidate the molecular mechanisms responsible for the anticancer property of Abnormal Savda Munziq. Materials and Methods. ASMq ethanol extract was prepared by a professional pharmacist. Caco-2 cells were treated with different concentration of ASMq ethanol extract (0.5–7.5 mg/mL for different time intervals (48 and 72 h. Antiproliferative effect of ASMq ethanol extract was determined by MTT assay; DNA fragmentation was determined by gel electrophoresis assay; cell cycle analysis was detected by flow cytometer; apoptosis-related gene expression was detected by RT-PCR assay. Results. ASMq ethanol extract possesses an inhibition effect on Caco-2 cells proliferation, induction of cell apoptosis, cell cycle arrest in sub-G1 phase, and downregulation of bcl-2 and upregulation of Bax gene expression. Conclusion. The anticancer mechanism of ASMq ethanol extract may be involved in antiproliferation, induction of apoptosis, cell cycle arrest, and regulation of apoptosis-related gene expression such as bcl-2 and Bax activity pathway.

  12. Preparation of poly-L-lysine functionalized magnetic nanoparticles and their influence on viability of cancer cells

    Science.gov (United States)

    Khmara, I.; Koneracka, M.; Kubovcikova, M.; Zavisova, V.; Antal, I.; Csach, K.; Kopcansky, P.; Vidlickova, I.; Csaderova, L.; Pastorekova, S.; Zatovicova, M.

    2017-04-01

    This study was aimed at development of biocompatible amino-functionalized magnetic nanoparticles as carriers of specific antibodies able to detect and/or target cancer cells. Poly-L-lysine (PLL)-modified magnetic nanoparticle samples with different PLL/Fe3O4 content were prepared and tested to define the optimal PLL/Fe3O4 weight ratio. The samples were characterized for particle size and morphology (SEM, TEM and DLS), and surface properties (zeta potential measurements). The optimal PLL/Fe3O4 weight ratio of 1.0 based on both zeta potential and DLS measurements was in agreement with the UV/VIS measurements. Magnetic nanoparticles with the optimal PLL content were conjugated with antibody specific for the cancer biomarker carbonic anhydrase IX (CA IX), which is induced by hypoxia, a physiologic stress present in solid tumors and linked with aggressive tumor behavior. CA IX is localized on the cell surface with the antibody-binding epitope facing the extracellular space and is therefore suitable for antibody-based targeting of tumor cells. Here we showed that PLL/Fe3O4 magnetic nanoparticles exhibit cytotoxic activities in a cell type-dependent manner and bind to cells expressing CA IX when conjugated with the CA IX-specific antibody. These data support further investigations of the CA IX antibody-conjugated, magnetic field-guided/activated nanoparticles as tools in anticancer strategies.

  13. Preparation of three-layered porous PLA/PEG scaffold: relationship between morphology, mechanical behavior and cell permeability.

    Science.gov (United States)

    Scaffaro, R; Lopresti, F; Botta, L; Rigogliuso, S; Ghersi, G

    2016-02-01

    Interface tissue engineering (ITE) is used to repair or regenerate interface living tissue such as for instance bone and cartilage. This kind of tissues present natural different properties from a biological and mechanical point of view. With the aim to imitating the natural gradient occurring in the bone-cartilage tissue, several technologies and methods have been proposed over recent years in order to develop polymeric functionally graded scaffolds (FGS). In this study three-layered scaffolds with a pore size gradient were developed by melt mixing polylactic acid (PLA) and two water-soluble porogen agents: sodium chloride (NaCl) and polyethylene glycol (PEG). Pore dimensions were controlled by NaCl granulometry while PEG solvation created a micropores network within the devices. Scaffolds were characterized from a morphological and mechanical point of view in order to find a correlation between the preparation method, the pore architecture and compressive mechanical behavior. Biological tests were also performed in order to study the effect of pore size gradient on the permeation of different cell lines in co-culture. To imitate the physiological work condition, compressive tests were also performed in phosphate buffered saline (PBS) solution at 37°C. The presented preparation method permitted to prepare three-layered scaffolds with high control of porosity and pore size distribution. Furthermore mechanical behaviors were found to be strongly affected by pore architecture of tested devices as well as the permeation of osteoblast and fibroblast in-vitro.

  14. Electrical properties of dye-sensitized solar cells prepared by the binder-free TiO2 pastes.

    Science.gov (United States)

    Park, Su-Ji; Lee, Jin; So, Soon-Youl; Hong, Kyung-Jin; Yun, Jung-Hyun; Kim, Sang-Ki

    2014-12-01

    Titania (TiO2) powder, which is material for photoelectrode in dye-sensitized solar cells (DSSCs), was fabricated by hydrothermal synthesis process at 230 degrees C for 12 hours. The crystal structures of all the synthesized nano-structured TiO2 films exhibited anatase phase. Binder-free pastes were prepared with the change of the amount of ammonia water from 2 μl to 640 μl in order to obtain the printable viscosity. It has been known that weak inter-particle bonds in slurry of flocculated particles make the slurry more viscous than slurry of dispersed particles. The increase of the amount of ammonia water for binder-free TiO2 pastes is attributed to the improvement of the viscosity of TiO2 paste and the power conversion efficiency of DSSCs using it. The viscosity of TiO2 pastes prepared at the ammonia water of 418 μl exhibited the highest value about 109,000 cP and also, was very transparent over 84%. As a result, the power conversion efficiency of DSSC prepared with the ammonia water of 418 μl was about 3%.

  15. Traditional Aboriginal Preparation Alters the Chemical Profile of Carica papaya Leaves and Impacts on Cytotoxicity towards Human Squamous Cell Carcinoma.

    Science.gov (United States)

    Nguyen, Thao T; Parat, Marie-Odile; Shaw, Paul N; Hewavitharana, Amitha K; Hodson, Mark P

    2016-01-01

    Carica papaya leaf decoction, an Australian Aboriginal remedy, has been used widely for its healing capabilities against cancer, with numerous anecdotal reports. In this study we investigated its in vitro cytotoxicity on human squamous cell carcinoma cells followed by metabolomic profiling of Carica papaya leaf decoction and leaf juice/brewed leaf juice to determine the effects imparted by the long heating process typical of the Aboriginal remedy preparation. MTT assay results showed that in comparison with the decoction, the leaf juice not only exhibited a stronger cytotoxic effect on SCC25 cancer cells, but also produced a significant cancer-selective effect as shown by tests on non-cancerous human keratinocyte HaCaT cells. Furthermore, evidence from testing brewed leaf juice on these two cell lines suggested that the brewing process markedly reduced the selective effect of Carica papaya leaf on SCC25 cancer cells. To tentatively identify the compounds that contribute to the distinct selective anticancer activity of leaf juice, an untargeted metabolomic approach employing Ultra High Performance Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry followed by multivariate data analysis was applied. Some 90 and 104 peaks in positive and negative mode respectively were selected as discriminatory features from the chemical profile of leaf juice and >1500 putative compound IDs were obtained via database searching. Direct comparison of chromatographic and tandem mass spectral data to available reference compounds confirmed one feature as a match with its proposed authentic standard, namely pheophorbide A. However, despite pheophorbide A exhibiting cytotoxic activity on SCC25 cancer cells, it did not prove to be the compound contributing principally to the selective activity of leaf juice. With promising results suggesting stronger and more selective anticancer effects when compared to the Aboriginal remedy, Carica papaya leaf juice warrants further study

  16. Preparation of triple-negative breast cancer vaccine through electrofusion with day-3 dendritic cells.

    Directory of Open Access Journals (Sweden)

    Peng Zhang

    Full Text Available Dendritic cells (DCs are professional antigen-presenting cells (APCs in human immune system. DC-based tumor vaccine has met with some success in specific malignancies, inclusive of breast cancer. In this study, we electrofused MDA-MB-231 breast cancer cell line with day-3 DCs derived from peripheral blood monocytes, and explored the biological characteristics of fusion vaccine and its anti-tumor effects in vitro. Day-3 mature DCs were generated from day-2 immature DCs by adding cocktails composed of TNF-α, IL-1β, IL-6 and PEG2. Day-3 mature DCs were identified and electofused with breast cancer cells to generate fusion vaccine. Phenotype of fusion cells were identified by fluorescence microscope and flow cytometer. The fusion vaccine was evaluated for T cell proliferation, secretion of IL-12 and IFN-γ, and induction of tumor-specific CTL response. Despite differences in morphology, day-3 and day-7 DC expressed similar surface markers. The secretion of IL-12 and IFN-γ in fusion vaccine group was much higher than that in the control group. Compared with control group, DC-tumor fusion vaccine could better stimulate the proliferation of allogeneic T lymphocytes and kill more breast cancer cells (MDA-MB-231 in vitro. Day-3 DCs had the same function as the day-7 DCs, but with a shorter culture period. Our findings suggested that day-3 DCs fused with whole apoptotic breast cancer cells could elicit effective specific antitumor T cell responses in vitro and may be developed into a prospective candidate for adoptivet immunotherapy.

  17. Competencies and Their Assessment

    Science.gov (United States)

    Drisko, James W.

    2014-01-01

    This article explores competencies and methods for their assessment in higher education and in social work's accreditation standards. Many contemporary policy and educational accreditation efforts employ the model of competency assessment. The current emphasis on accountability in higher education, including the Council on Social Work…

  18. Analyzing ADN competencies.

    Science.gov (United States)

    Mitchell, P R

    1989-01-01

    El Paso Community College District, using the DACUM Process, identified 19 major competency areas with 313 specific competencies for AD Nursing. This article provides an overview of the DACUM Process, a discussion of the application to the ADN program, a summary of the results, and future activities.

  19. Cultural Competence Revisited

    Science.gov (United States)

    Garran, Ann Marie; Werkmeister Rozas, Lisa

    2013-01-01

    In 2001, the National Association of Social Workers (NASW) adopted 10 discrete standards of culturally competent practice which undergird our commitment to diversity and social justice. The concept of intersectionality is newly emerging in social work, though, causing us to reflect on our current conceptualizations of cultural competence.…

  20. Intercultural Competence Teaching

    Institute of Scientific and Technical Information of China (English)

    HUI Ran

    2015-01-01

    It is widely realized that cultural competence is playing an increasingly important role in language teaching because of the close associations between culture and language. To enhance the efficiency of language teaching, this essay will elucidate the importance of intercultural competence teaching and introduce three methods of cultural teaching.

  1. Competence, Curriculum, and Control.

    Science.gov (United States)

    Jackson, Nancy S.

    1988-01-01

    Draws upon a case study of a community college program review to examine the application of a competency-based approach to the process of curriculum design. Suggests that competency-based curriculum development shifts the basis for decision making from teacher knowledge to an objectified accounting system of employers and curriculum technicians.…

  2. Validating clinical competence.

    Science.gov (United States)

    Chappell, Kathy; Koithan, Mary

    2012-07-01

    Professional registered nurses must be competent to provide care in today's fast-paced, highly technical clinical environment. Competency programs designed to teach and evaluate essential knowledge, skills, and abilities of registered nurses are one method organizations may use to demonstrate nursing proficiency.

  3. Competencies in Ornamental Horticulture

    Science.gov (United States)

    Loewen, Curtis E.

    1974-01-01

    Based on the author's dissertation, this article pertains to the identification of competencies for ornamental horticulture workers in Oregon. Findings were based on interviews with 56 ornamental horticulture business employers regarding 100 competencies. The method used can serve as a model for obtaining occupational information to develop and…

  4. Developing Clinical Competence

    NARCIS (Netherlands)

    P.F. Wimmers (Paul)

    2006-01-01

    textabstractThe development of clinical competence is the main purpose of medical education. The long road to become clinically competent starts on the first day of medical school, and every institution strives to select the best students. The responsibility of medical schools is to train th

  5. The Spiritual Competency Scale

    Science.gov (United States)

    Robertson, Linda A.

    2010-01-01

    This study describes the development of the Spiritual Competency Scale, which was based on the Association for Spiritual, Ethical and Religious Values in Counseling's original Spiritual Competencies. Participants were 662 counseling students from religiously based and secular universities nationwide. Exploratory factor analysis revealed a 22-item,…

  6. Assessment of Innovation Competency

    DEFF Research Database (Denmark)

    Nielsen, Jan Alexis

    2015-01-01

    The author employed a 3-step qualitative research design with multiple instances of source validation to capture expert teachers’ (n = 28) reflections on which manifest signs they would look for when they asses students’ innovation competency. The author reports on the thematic analysis of the re......The author employed a 3-step qualitative research design with multiple instances of source validation to capture expert teachers’ (n = 28) reflections on which manifest signs they would look for when they asses students’ innovation competency. The author reports on the thematic analysis...... of the recorded talk in interaction that occurred in teacher group discussion sessions at 5 upper secondary schools. Based on the analysis, it was possible to extrapolate assessment criteria for 5 subcompetencies relevant to innovation (creative competency, collaboration competency, navigation competency, action...... competency, and communication competency) as well as assessment criteria for a number of skills relevant to these subcompetencies. These assessment criteria, it is argued, largely resonate with existing literature and they provide a detailed glimpse into how assessment of innovation competency could...

  7. Developing Clinical Competence

    NARCIS (Netherlands)

    P.F. Wimmers (Paul)

    2006-01-01

    textabstractThe development of clinical competence is the main purpose of medical education. The long road to become clinically competent starts on the first day of medical school, and every institution strives to select the best students. The responsibility of medical schools is to train

  8. CREATIVE PRACTICES IN DEVELOPING INTERCULTURAL COMMUNICATIVE COMPETENCE

    Directory of Open Access Journals (Sweden)

    Lyudmila M. Andryukhina

    2016-09-01

    Full Text Available Introduction: the article is concerned with the topical problem of extending intercultural communicative competence in the educational process. Materials and Methods: the authors made a theoretical and empirical analysis of domestic and foreign research and analytical reports prepared in the framework of international projects, the activity of non-governmental organisations, in particular the activities of Sverdlovsk regiona l public organisation “Friends of France”. Results: it is proven that intercultural competence cannot be fully developed by traditional forms of education, and even the existing current innovative experience in this area is insufficient. The integration of education becomes a crucial factor. The article describes the creative educational practices of such integration as the conditions for broadening intercultural communicative competence. The basic characteristics of creative practices are described. Сreative practices being integrated fo rm a creative platform. Discussion and Conclusions: the system of conditions and means for broadening intercultural communicative competence is in constant development. The authors propose to consider the communicative competence as a system of creative practices transformed in the course of elaboration into creative platforms, expanding in the process of intercultural dialogue. This is a promising way to achieve the comprehensive result – completeness of integrated multi-component structure of the intercultural co mmunicative competence.

  9. Efficient Light Harvester Layer Prepared by Solid/Mist Interface Reaction for Perovskite Solar Cells.

    Science.gov (United States)

    Xia, Xiang; Li, Hongcui; Wu, Wenyi; Li, Yanhua; Fei, Dehou; Gao, Chunxiao; Liu, Xizhe

    2015-08-12

    A solid/mist reaction method is developed to produce well-crystallized light harvester layers without pinhole defects for perovskite solar cells. The reaction based on mist precursor can be facilely operated with low process temperature. And it can effectively control the volume of CH3NH3I solution and the reaction temperature, which affect the quality of perovskite harvester layers and the performance of perovskite solar cells remarkably. Under optimized condition, the efficiencies of devices reach 16.2% with the average efficiency of 14.9%. The solid/mist reaction is also used to fabricate planar junction solar cells and a PCE of 14.9% is obtained.

  10. 40% Efficiency enhancement in solar cells using ZnO nanorods as shell prepared via novel hydrothermal synthesis

    Science.gov (United States)

    Ebadi, Mohammad; Zarghami, Zabihullah; Motevalli, Kourosh

    2017-03-01

    Herein, rod-like ZnO nanostructures were synthesized via a novel hydrothermal route using Zn(OAc)2, ethylenediamine and hydrazine as a new set of starting reagents. The as-synthesized products were characterized by techniques including XRD, EDS, SEM, XPS, Pl and FTIR. The prepared ZnO nanostructures were utilized as shell on TiO2 film in DSSCs. Effect of precursor type, morphology and thickness of ZnO shell (number of electrophoresis cycle) on solar cells efficiency were well studied. Our results showed that ethylenediamine has crucial effect on morphology of synthesized ZnO nanostructures and using ZnO nanostructures leads to an increase in DSSCs efficiency compared to bare TiO2 from 4.66 to 7.13% ( 40% improvement). Moreover, highest amount of solar cell efficiency (7.13%) was obtained by using ZnO nanorods with two cycle of electrophoresis for deposition.

  11. Stabilization of polymer lipid complexes prepared with lipids of lactic acid bacteria upon preservation and internalization into eukaryotic cells.

    Science.gov (United States)

    Alves, P; Hugo, A A; Szymanowski, F; Tymczyszyn, E E; Pérez, P F; Coelho, J F J; Simões, P N; Gómez-Zavaglia, A

    2014-11-01

    The physicochemical characterization of polymer liposome complexes (PLCs) prepared with lipids of lactic acid bacteria and poly(N,N-dimethylaminoethyl methacrylate) covalently bound to cholesterol (CHO-PDMAEMA) was carried out in an integrated approach, including their stability upon preservation and incorporation into eukaryotic cells. PLCs were prepared with different polymer:lipid molar ratios (0, 0.05 and 0.10). Zeta potential, particle size distribution and polydispersity index were determined. The optimal polymer:lipid ratio and the stability of both bare liposomes and PLCs were evaluated at 37 °C and at different pHs, as well as after storage at 4 °C, -80 °C and freeze-drying in the presence or absence of trehalose 250 mM. Internalization of PLCs by eukaryotic cells was assessed to give a complete picture of the system. Incorporation of CHO-PDMAEMA onto bacterial lipids (ratio 0.05 and 0.10) led to stabilization at 37 °C and pH 7. A slight decrease of pH led to their strong destabilization. Bacteria PLCs showed to be more stable than lecithin (LEC) PLCs (used for comparison) upon preservation at 4 and -80 °C. The harmful nature of the preservation processes led to a strong decrease in the stability of PLCs, bacterial formulations being more stable than LEC PLCs. The addition of trehalose to the suspension of liposomes stabilized LEC PLC and did not have effect on bacterial PLCs. In vitro studies on Raw 264.7 and Caco-2/TC7 cells demonstrated an efficient incorporation of PLCs into the cells. Preparations with higher stability were the ones that showed a better cell-uptake. The nature of the lipid composition is determinant for the stability of PLCs. Lipids from lactic acid bacteria are composed of glycolipids and phospholipids like cardiolipin and phosphatidylglycerol. The presence of negatively charged lipids strongly improves the interaction with the positively charged CHO-PDMAEMA, thus stabilizing liposomes. In addition, glycolipids and

  12. Dye-Sensitized Solar Cells with Anatase TiO2 Nanorods Prepared by Hydrothermal Method

    OpenAIRE

    Ming-Jer Jeng; Yi-Lun Wung; Liann-Be Chang; Lee Chow

    2013-01-01

    The hydrothermal method provides an effective reaction environment for the synthesis of nanocrystalline materials with high purity and well-controlled crystallinity. In this work, we started with various sizes of commercial TiO2 powders and used the hydrothermal method to prepare TiO2 thin films. We found that the synthesized TiO2 nanorods were thin and long when smaller TiO2 particles were used, while larger TiO2 particles produced thicker and shorter nanorods. We also found that TiO2 films ...

  13. Cumulus cell transcripts transit to the bovine oocyte in preparation for maturation

    DEFF Research Database (Denmark)

    Macaulay, Angus D.; Gilbert, Isabelle; Scantland, Sara

    2016-01-01

    , revealed transcripts common to all three fractions, suggesting the use of transferred transcripts for translation. Furthermore, the removal of potential RNA trafficking by stripping the cumulus cells caused a significant reduction in maturation rates, indicating the need for the cumulus cell RNA transfer...... that the somatic cells also contribute transcripts to the gamete. Here, we show that these transcripts can be visualized moving down the transzonal projections (TZPs) to the oocyte, and that a time course analysis revealed progressive RNA accumulation in the TZPs, indicating that RNA transfer occurs before......So far, the characteristics of a good quality egg have been elusive, similar to the nature of the physiological, cellular, and molecular cues leading to its production both in vivo and in vitro. Current understanding highlights a strong and complex interdependence between the follicular cells...

  14. Influence of electrospun scaffolds prepared from distinct polymers on proliferation and viability of endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Matveeva, V. G., E-mail: matveeva-vg@mail.ru; Antonova, L. V., E-mail: antonova.la@mail.ru; Velikanova, E. A.; Sergeeva, E. A.; Krivkina, E. O.; Glushkova, T. V.; Kudryavtseva, Yu. A.; Barbarash, O. L.; Barbarash, L. S. [Research Institute for Complex Issues of Cardiovascular Diseases, Kemerovo, 650002 (Russian Federation)

    2015-10-27

    We compared electrospun nonwoven scaffolds from polylactic acid (PLA), polycaprolactone (PCL), and polyhydroxybutyrate/valerate (PHBV)/polycaprolactone (PHBV/PCL). The surface of PHBV/PCL and PCL scaffolds was highly porous and consisted of randomly distributed fibers, whilst the surface of PLA scaffolds consisted of thin straight fibers, which located more sparsely, forming large pores. Culture of EA.hy 926 endothelial cells on these scaffolds during 7 days and further fluorescent microscopy demonstrated that the surface of PHBV/PCL scaffolds was most favorable for efficient adhesion, proliferation, and viability of endothelial cells. The lowest proliferation rate and cell viability were detected on PLA scaffolds. Therefore, PHBV/PCL electrospun nonwoven scaffolds demonstrated the best results regarding endothelial cell proliferation and viability as compared to PCL and PLA scaffolds.

  15. mc-Si:H/c-Si solar cell prepared by PECVD

    Institute of Scientific and Technical Information of China (English)

    XU Ying; LIAO Xianbo; DIAO Hongwei; Li Xudong; ZENG Xiangbo; LIU Xiaoping; WANG Minhua; WANG Wenjing

    2006-01-01

    Hetero-junction solar cells with an mc-Si:H window layer were achieved. The open voltage is increased while short current is decreased with increasing the mc-Si:H layer's thickness of emitter layer. The highest of V oc of 597 mV has obtained. When fixed the thickness of 30 nm, changing the N type from amorphous silicon layer to micro-crystalline layer, the efficiency of the hetero-junction solar cells is increased. Although the hydrogen etching before deposition enables the c-Si substrates to become rough by AFM images, it enhances the formation of epitaxial-like micro-crystalline silicon and better parameters of solar cell can be obtained by implying this process. The best result of efficiency is 13.86% with the V oc of 549.8 mV, J sc of 32.19 mA·cm-2 and the cell's area of 1cm2.

  16. Intercultural Teaching Competence: A Multi-Disciplinary Model for Instructor Reflection

    Science.gov (United States)

    Dimitrov, Nanda; Haque, Aisha

    2016-01-01

    This article presents a model for Intercultural Teaching Competence (ITC) that instructors may use as a tool for reflection as they prepare to facilitate learning across cultures. Building on previous research on intercultural competence, culturally relevant teaching, intercultural trainer competencies, and student-centred approaches to teaching,…

  17. The Competency-Based Movement in Student Affairs: Implications for Curriculum and Professional Development

    Science.gov (United States)

    Eaton, Paul William

    2016-01-01

    This article examines the limitations and possibilities of the emerging competency-based movement in student affairs. Using complexity theory and postmodern educational theory as guiding frameworks, examination of the competency-based movement will raise questions about overapplication of competencies in graduate preparation programs and…

  18. Profesional Competencies among Pre-Service Teachers in Special Education from Their Perspectives

    Science.gov (United States)

    Theeb, Raied Sheikh; Muhaidat, Mohammad Ali; Al-Zboon, Eman Khleif

    2014-01-01

    Due to the contemporary educational reform, the movement of teachers' preparation based on the competencies became the most prominent field of study. The current study aimed at exploring Professional Competencies among special education pre-service teachers (N = 112) and the level of their competencies' practice from their perspectives.…

  19. Identifying Transition Teacher Competencies through Literature Review and Surveys of Experts and Practitioners

    Science.gov (United States)

    Morgan, Robert L.; Callow-Heusser, Catherine A.; Horrocks, Erin L.; Hoffmann, Audrey N.; Kupferman, Scott

    2014-01-01

    We first conducted a synthesis of literature to identify essential transition teacher competencies to guide curriculum development for a personnel preparation program. The synthesis yielded a list of 67 competencies needed by transition teachers. Using the 67 competencies, we next created an electronic survey in which respondents were asked to…

  20. Competencies in nursing students for organized forms of clinical moral deliberation and decision-making

    NARCIS (Netherlands)

    Uil-Westerlaken, Jeanette den; Cusveller, Bart

    2013-01-01

    Bachelor-prepared nurses are expected to be competent in moral deliberation and decision-making (MDD) in clinical practice. It is unclear, however, how this competence develops in nursing students. This study explores the development of nursing students’ competence for participating in organized for