Journal articles on the topic 'Cultures DIY'
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1
Chrysagis, Evangelos. "DIY Cultures and Underground Music Scenes." Popular Music and Society 42, no. 5 (August 5, 2019): 624–26. http://dx.doi.org/10.1080/03007766.2019.1650522.
2
Bennett, Andy. "Conceptualising the Relationship Between Youth, Music and DIY Careers: A Critical Overview." Cultural Sociology 12, no. 2 (February 19, 2018): 140–55. http://dx.doi.org/10.1177/1749975517750760.
Abstract:
The purpose of this article is to explore how creative competencies acquired through involvement in music and style-based youth cultures are being converted by young people into DIY (do-it-yourself) careers. This term, as applied here, covers a range of alternative and do-it-yourself modes of work and employment not governed by the acquisition of formal qualifications and training, but, rather, grounded in knowledge and practical know-how acquired through participation in music and style-based youth cultures and associated consumption, leisure and lifestyle practices. The article examines how the emergence of the DIY career can be placed in the context of the collapse of the youth labour market and the increasingly precarious position in which young people find themselves. It also argues that, given the increasing emphasis among youth (and indeed post-youth individuals) placed on the significance of music as a source of DIY career-making, some redefinition of the term DIY music-making and the sphere of DIY creative practice is necessary, particularly in respect of its relationship to more mainstream cultural industries and the related concept of the ‘creative city’.
3
Guerra, Paula. "Raw Power: Punk, DIY and Underground Cultures as Spaces of Resistance in Contemporary Portugal." Cultural Sociology 12, no. 2 (April 16, 2018): 241–59. http://dx.doi.org/10.1177/1749975518770353.
Abstract:
This article focuses on presenting, analysing and understanding the ‘do-it-yourself’ (DIY) representations of over 200 participants in Portuguese punk music scenes between 1977 and 2014. Through an ethnographic study of punk and the DIY ethos in Portugal, the article depicts its local interpretation and significance, and highlights the singularities of this socio-historical context. Our approach has three focal points: (1) the importance of DIY punk manifestations in the development of youth (sub)cultures, including forms of production and consumption of music, fashion, aesthetics, leisure, the night and cosmopolitanism; (2) the singularity of DIY punk manifestations in Portugal and related resistance practices in their correlation with social, political and economic development in a country outside the Anglo-Saxon context; and (3) the embedding of a DIY ethos and the associated claim to authenticity in the professions and careers in which punks engage today.
4
Threadgold, Steven. "Creativity, Precarity and Illusio: DIY Cultures and ‘Choosing Poverty’." Cultural Sociology 12, no. 2 (October 25, 2017): 156–73. http://dx.doi.org/10.1177/1749975517722475.
Abstract:
Young people investing themselves in DIY cultures have to negotiate the complex but now normalised nexus of employment, unemployment and underemployment to make ends meet, while maintaining space in their lives to pursue their creative and artistic passions. This article presents research with young people in an underground music scene across Australia who are balancing economic pressures with their desires to generate and uphold DIY and punk-influenced activities in a networked community of likeminded friends and collaborators. Many of the participants actively ‘choose poverty’, that is, they knowingly and strategically make decisions that ‘keep overheads low’ to free up temporal and mental space to continue to be creative. Their ideas of being successful are not expressed in material terms, but are contingent upon a future where they continue to have the opportunity to invest themselves in their interests even if that means living in relative poverty. Using the oft-ignored Bourdieusian concepts of illusio, struggle and strategy, this article provides a case study of some of the ways young people deal with the risks and opportunities of a precarious existence. In this case, living an ethical life trumps material concerns, projecting a hopeful attitude towards the future.
5
Kim, Min-Hyoung. "A Semiotic Adaptation of Materiality Studies: With a Focus of DIY Making Cultures." Semiotic Inquiry 72 (December 30, 2022): 7–37. http://dx.doi.org/10.24825/si.72.1.
6
Haenfler, Ross. "The Entrepreneurial (Straight) Edge: How Participation in DIY Music Cultures Translates to Work and Careers." Cultural Sociology 12, no. 2 (June 27, 2017): 174–92. http://dx.doi.org/10.1177/1749975517700774.
Abstract:
Only recently have researchers begun thoroughly examining the role of youth music cultures and subcultures in participants’ ‘adult’ lives, suggesting that participation does not end with an abrupt transition to adulthood. Significantly, how subcultural experience translates into work skills and job opportunities needs further investigation. Based upon interviews and participant observation with older straight edgers – clean-living punks associated with the hardcore music scene – over the course of five years, this article examines subcultural entrepreneurs, in particular straight edgers, who have used DIY experiences to start businesses or advance careers. I first examine how involvement in DIY scenes can influence choice of career; many participants chose work reflective of their DIY values and/or created their own career. Next, I discuss how participants’ DIY dispositions influence their work, including tolerance for risk, confidence, self-reliance, and adaptability. I then demonstrate how participants translate scene experience into useful and marketable skills, including management, organization, marketing, and job-specific proficiencies.
7
Guerra, Paula. "So close yet so far: DIY cultures in Portugal and Brazil." Cultural Trends 30, no. 2 (January 24, 2021): 122–38. http://dx.doi.org/10.1080/09548963.2021.1877085.
8
Cleveston Gelain, Gabriela, Milene Migliano, and Pedro De Assis Pereira Scudeller. "Experiências de uma Riot Grrrl: Kathleen Hanna, feminismo, DIY e cultura remix." Revista PHILIA | Filosofia, Literatura & Arte 2, no. 2 (November 10, 2020): 152–88. http://dx.doi.org/10.22456/2596-0911.104017.
Abstract:
Por meio dos fragmentos de narrativas da trajetória da musicista e ativista Kathleen Hanna, pioneira do movimento Riot Grrrl, remontamos, a partir do documentário The Punk Singer, a terceira onda do movimento feminista, evidenciando a interseccionalidade e protagonismo juvenil. Através de fanzines, arte, colagens, letras de música, performances e formação de bandas a partir de uma filosofia punk do it yourself (DIY), salientamos a ampla contribuição de Hanna para o feminismo contemporâneo ao desafiar um cenário opressor dentro do movimento punk, estimulando o surgimento de outras iniciativas feministas, rebeldes e riot grrrls. Sua prática e performance artística são abordadas pelo viés das culturas DIY e remix, potencializando partilhas do sensível e politicidades no engajamento de subjetividades que superam a contenção dos imaginários vigentes.Palavras-chave: Feminismo. Riot Grrrl. Kathleen Hanna. Cultura remix. Imaginário político. AbstractThrough fragments of narratives gathered from the documentary The Punk Singer, based on the life and career of musician and activist Kathleen Hanna, pioneer of the Riot Grrrl movement, we refer to the third wave of the feminist movement, by demonstrating the dimensions of intersectionality and youth protagonism within her work. Ranging from fanzines, art, collages and lyric-making to performance and music groups based on a punk "do it yourself" (DIY) philosophy, we highlight Hanna's broad contribution to contemporary feminism by challenging an oppressive scenario within the punk movement, and by stimulating the emergence of other feminist, rebel and riot grrrls initiatives. Her artistic practice and performance are analyzed through the bias of DIY and remix cultures, thus potentializing distributions of the sensible and politicities in the engagement of subjectivities that surpass the containment of current imaginaries.Keywords: Feminism. Riot Grrrl. Kathleen Hanna. Remix culture. Political imaginary.
9
Barnard, Katharine D., Ralph Ziegler, David C. Klonoff, Katarina Braune, Bettina Petersen, Til Rendschmidt, Daniel Finan, Aaron Kowalski, and Lutz Heinemann. "Open Source Closed-Loop Insulin Delivery Systems: A Clash of Cultures or Merging of Diverse Approaches?" Journal of Diabetes Science and Technology 12, no. 6 (August 6, 2018): 1223–26. http://dx.doi.org/10.1177/1932296818792577.
Abstract:
Biomedical outcomes for people with diabetes remain suboptimal for many. Psychosocial care in diabetes does not fare any better. “Artificial pancreas” (also known as “closed-loop” and “automated insulin delivery”) systems present a promising therapeutic option for people with diabetes (PWD)—simultaneously improving glycemic outcomes, reducing the burden of self-management, and improving health-related quality of life. In recent years there has emerged a growing movement of PWD innovators rallying behind the mantra #WeAreNotWaiting, developing “do-it-yourself artificial pancreas systems (DIY APS).” Self-reported results by DIY APS users show improved metabolic outcomes such as impressive stability of glucose profiles, significant reduction of A1c, and more time within their glycemic target range. However, the benefits remain unclear for the broader population of PWD beyond these highly engaged, highly tech-savvy users willing and able to engage in the demands of building and maintaining their DIY APS. We discuss the challenges faced by key stakeholder groups in terms of potential collaboration and open debate of these challenges.
10
Critchley, Tom. "DIY Design and Radical Worldbuilding at The Grove Skatepark, London." Temes de Disseny, no. 39 (July 27, 2023): 208–27. http://dx.doi.org/10.46467/tdd39.2023.208-227.
Abstract:
This paper presents an ethnographic case study of DIY design practices at a skatepark in London, UK. The skatepark is presented as an urban commons, offering a framework for DIY infrastructuring that contrasts top-down and inequitable urbanisation processes. Three key themes emerge from the fieldwork that demonstrate the capacities and limitations of DIY design. Firstly, DIY practices are viewed as “learning-by-doing”, wherein skateboarders not only learn how to construct a skatepark but also an array of vocational and soft skills. Here, DIY’s qualities of low barriers to participation, multidisciplinarity, and low risk of failure foreground a rich environment of vernacular knowledge exchange that underpins the research methodology of this paper utilising skatepark construction as object-orientated, research-through-design. Secondly, DIY design is found to support a community of practitioners that transcend the space as just a skatepark, incorporating a community garden, collaborative theatre project and arts-based workshops that transcend male-dominated histories of DIY skateboarding cultures. Here, Participatory Design (PD) notions of ‘infrastructuring’ mirror an emergent and open-ended design process that supports inclusive socio-material worldmaking. Thirdly, DIY design practices influence the ways in which the space is governed, in which there are found to be contradictory notions of “prefigurative politics.” Here, the space is argued to be inclusive and anarchic yet centres agency around notions of “core skateboarding” that reinforce masculine hegemonies. Collectively, this paper argues that DIY design has the potential to serve as a worldmaking agent underpinned by a unique set of values and modes which are well-suited to times of uncertainty, flux and crisis, yet highlights the necessity for critically engaging with the politics and practices of both DIY and skateboarding communities.
11
Foster, Ellen K. "Claims of Equity and Expertise: Feminist Interventions in the Design of DIY Communities and Cultures." Design Issues 35, no. 4 (September 2019): 33–41. http://dx.doi.org/10.1162/desi_a_00562.
Abstract:
Using both interviews and participant observations, this article examines feminist design initiatives in hacker and maker cultures, unpacking their possible contribution to a world of more equitable digital technology development, use, and re(de)construction.
12
Virtù, Ludovico V. "Displacing the Gender Binary Through Modes of Dis/Organizing: Sex Toys, Sexuality and Trans Politics." Politics and Governance 8, no. 3 (September 18, 2020): 321–31. http://dx.doi.org/10.17645/pag.v8i3.3016.
Abstract:
Scholars in sexuality and organization studies have highlighted the centrality of sexuality in organizational power and the ways in which sexuality is in/visibilized, controlled, violently exercised, normativized, and/or resisted in organizations. However, there is still little empirical research focusing on social-movement organizations that promote political change in transgender sexual cultures. With this article, I contribute a qualitative case study of a trans and non-binary do-it-yourself (DIY) sex-toy workshop. Drawing on organization, social-movement, and transgender studies, I develop the notion of ‘trans-organizing’ as a specific mode of organizing and ask: How does trans-organizing around sexuality displace the gender binary in the context of a DIY sex-toy workshop? My findings hint at three dis/organizing processes: dis/organizing language, embodiment, and knowledge sharing.
13
Rokhim, Nur. "Tradisi Shalawat Ngelik di Kampung Santri Mlangi, Sleman, DIY." Tamaddun: Jurnal Kebudayaan dan Sastra Islam 20, no. 1 (June 28, 2020): 15–28. http://dx.doi.org/10.19109/tamaddun.v20i1.5951.
Abstract:
The tradition of ngelik prayer is one of the traditions of the celebration of the birth of the Prophet Muhammad, in the Santri Village in Mlangi, Sleman, DIY. This tradition is carried out at the peak of the prophet's birthday celebration which is the 12th of Rabiu'ul Awal at Pathok Negoro Mosque in Mlangi. This research tries to reveal in detail the tradition and try to analyze why this tradition still exists today. This study uses two data collection techniques, namely literature and field studies. The approach used is a qualitative approach with an ethnographic model. This research reveals a brief history of Santri Mlangi Village formed by the initiative of KH. Nur Iman Mlangi, sibling of Sultan Hamengkubuwono I. This research also found that the tradition of ngelik prayer is a new culture as a result of a cross between Islamic and Javanese cultures. The celebration of the birthday of the Prophet Muhammad SAW, is an Islamic culture. While recitation of shalawat using Javanese-style notes is a Javanese tradition. This is inseparable from the influence of the existence of KH. Nur Iman Mlangi who is a royal descendant of the Yogyakarta Palace. This study also revealed that the existence of dozens of pesantren has a big role in maintaining the continuity of the tradition of ngelik prayer.
Keywords: Shalawat Ngelik, Mlangi, Pathok Negoro Mosque
14
Danilova, Anna G. "QUANTITATIVE METHODS IN THE HISTORICAL AND PSYCHOLOGICAL STUDY INTO THE CATEGORICAL STRUCTURE OF VALUES AND IMAGES IN DIFFERENT CULTURES." Moscow University Psychology Bulletin, no. 1 (2023): 221–46. http://dx.doi.org/10.11621/vsp.2023.01.10.
Abstract:
Background. X e growing interest in the historical development of social systems requires an assessment of the possibilities for a quantitative comparison of psychological characteristics in a person and society of diY erent eras. Objective.X e study aims to reveal the structure of social values and features of contemporary images according to verbal and visual texts of the cultures in the past.Methods. Behavior characteristics describing G. Hofstede’s basic cultural dimensions (Hofstede, 1980) were used to encode texts. Psychosemantic assessment of the works of art perception was carried out with the use of a scale based on the multifaceted Five Factor Model of personality by P. Costa, R. McCrae (FFM; NEO-PI-R questionnaire adapted by V.E. Orel et al.) and the scale of individual image perception in media communication by L.V. Matveeva et al. Sample. A) Ancient Greek and Roman texts of 7th c. BC — 4th c. AD, Old Russian texts of 11th–17th centuries, Byzantine texts of 5th–15th centuries and texts of medieval European culture according to the 17th c.— a total of 205 separate texts. B) Visual stimulus material on cultures: Ancient Greece (7th–1th c. BC); Ancient Rome (7th c. BC — 4th c. AD); Byzantium (5th–15th cc.); Russia (11–19 cc.); Europe (9th, 11th–17th cc.) was assessed by respondents of diY erent ages from 14 to 70 (N = 68). Results. Reliability analysis and confirmatory factor analysis generally supported the structure of the W ve-factor model of personality and the basic dimensions of culture by G. Hofstede on the material of historical texts and works of art. X e least stable dimensions are Hofstede’s Masculinity and Openness to Experience factors of the personality model. An exploratory factorial analysis of data on historical texts yielded a factor structure reZ ecting speciW c aspects of Hofstede’s dimensions. In general, the correlations between personality traits and basic cultural dimensions described by G. Hofstede and R. McCraе (Хофстеде, МакКрэй, 2010) are reproduced. X e diY erences mainly relate to the FFM factor “Extraversion”. X e features of social situations described in historical narratives that are not W xed by the characteristics of behavior that describe the basic dimensions of culture according to Hofstede are discussed. Conclusion. X e subdimensions of modern models of personality and cultural values make it possible to obtain a fairly adequate quantitative assessment of the cultures of the past.
15
Oswal, Sushil K., and Zsuzsanna B. Palmer. "Culturally Situated Do-It-Yourself Instructions for Making Protective Masks: Teaching the Genre of Instructional Design in the Age of COVID-19." Journal of Business and Technical Communication 35, no. 1 (September 18, 2020): 160–66. http://dx.doi.org/10.1177/1050651920959190.
Abstract:
This article employs cross-cultural communication approaches to teaching instructional design in the times of COVID-19 pandemic. Focusing on instructions from France, India, Spain, and the United States for making protective masks, the authors highlight how the writers and designers of these four documents from each culture approach their audiences, organize their DIY instructions, make language choices, employ images and other illustration devices, and culturally persuade users. While acknowledging cultural differences, the authors urge students to identify and adopt design strengths from diverse cultures in their own ideas about composing instructions.
16
Pavlidis, Adele, and Wendy O’Brien. "Sport and feminism in China: On the possibilities of conceiving roller derby as a feminist intervention." Journal of Sociology 53, no. 3 (July 29, 2017): 704–19. http://dx.doi.org/10.1177/1440783317721349.
Abstract:
The spread of contemporary roller derby presents an opportunity to examine the ways sport can act as a form of feminist intervention. This article draws on a qualitative case study of a roller derby league in China, made up predominantly of expatriate workers, to explore some of the possibilities roller derby presents in activating glocal forms of feminist participatory action. The globalization of sport has often been associated with colonialism and the loss of local physical cultures, together with commercialization. Roller derby provides a very different case, where, together with the spread of the game, comes a focus on gender diversity, female strength and a particular derby style of DIY (do-it-yourself) governance that may support forms of participatory action. As an exploratory study, this article points towards the potential of roller derby, and possibly other sport cultures, to support human rights activism in the Asia Pacific.
17
Guerra, Paula. "‘Just can’t go to sleep’: DIY cultures and alternative economies from the perspective of social theory." Portuguese Journal of Social Science 16, no. 3 (September 1, 2017): 283–303. http://dx.doi.org/10.1386/pjss.16.3.283_1.
18
Silva, Gabriel Barth da. "As reconfigurações das memórias. Uma resenha do livro Punk, Fanzines and DIY Cultures in a Global World." Todas as Artes Revista Luso-Brasileira de Artes e Cultura 3, no. 3 (2020): 130–35. http://dx.doi.org/10.21747/21843805/tav3n3r1.
19
Stage, Carsten, and Sophie Esmann Andersen. "Ambiguous Imitations: DIY Hijacking the ’Danish Mother Seeking’ Stealth Marketing Campaign on YouTube." Culture Unbound 4, no. 2 (June 1, 2012): 393–414. http://dx.doi.org/10.3384/cu.2000.1525.124393.
Abstract:
The purpose of this paper is to explore the concept of imitation as a key dimension of online DIY and participatory cultures on YouTube. The empirical point of departure is the viral stealth marketing YouTube video entitled ’Danish Mother Seeking’, produced by the official national tourist organisation (Visit Denmark), and selected extracts of the online responses to this video. Framed by the notion of participatory culture (Jenkins 2006; Burgess & Green 2009) and the concept of imitation (Tarde 1895/1903), we analyse how marketing initiatives buy into and borrow energy from engaged networked produsers, but also how these produsers can criticise marketing initiatives by ’re-imitating’ them. Following this, we argue that the case represents an interesting and fascinating example of consumer re-sistance and bottom-up voices insisting on being heard, rather than a simple example of the breakdown of a brand strategy. Looking at the response videos they furthermore reveal that imitation can be a rather ambiguous social strategy as it is used both to transfer energy from the imitated object and to deconstruct it. As part of this argument we replace the classical concept of ’mimicry’ (Bhabha 1994) with the notion of ’ambiguous imitation’ to be able to describe online imitation as both an act of critical voicing and energy transmission.
20
Wicaksono, Raden Ichlasul Zinedine Bieldam, Putu Agus Wikanatha Sagita, and NGAS Dewi. "PENGEMASAN PAKET WISATA BERBASIS BUDAYA DENGAN PEMANFAATAN MODA TRANSPORTASI TRADISIONAL ANDONG DI KOTA YOGYAKARTA." Jurnal IPTA 10, no. 1 (July 28, 2022): 32. http://dx.doi.org/10.24843/ipta.2022.v10.i01.p04.
Abstract:
Yogyakarta is the largest number of tourists in DIY Province. Focuses of tourism development sector is cultural tourism, one of the cultures owned by the city of Yogyakarta is andong traditional transportation which is starting to lose competition with modern transportation. Currently, Yogyakarta Tourism Department is focusing on increasing the number of visits and length of stay of tourists with various efforts, which is tour packages. Therefore, the packaging of cultural tourism packages using andong needs to be done to anticipate the problems of traditional transportation and to increase the number of tourist visits. Data collection techniques used are observation, interviews, documentation and literature study. Informant determination technique used was purposive sampling with Tourism Department and Transportation Department of Yogyakarta as base informant and Andong DIY Association and manager of cultural tourism destinations as key informants. Analytical technique used is descriptive qualitative. Based on the research results, the city of Yogyakarta has the potential for cultural tourism in 5 indicators such as arts exhibition (Keraton Yogyakarta), arts (Taman Pintar and Sonobudoyo Museum), festivals (Tuesday Wage Art Festival), traditional food (Gudeg Yu Djum and Bakpia Pathok 25) and history (Fort Vredeburg Museum and Taman Sari). Andong has important components in packaging tour packages such as conditions and facilities, price/cost, time, location, accessibility and uniqueness. The potential for cultural tourism will be packaged by 3 tour packages, namely Chronicle of Kasultanan Ngayogyakarta, Art of Yogyakarta and Countdown Tuesday Wage. The package lasts for halfday and will be managed by Andong DIY Association.
21
Reitsamer, Rosa, and Rainer Prokop. "Keepin’ it Real in Central Europe: The DIY Rap Music Careers of Male Hip Hop Artists in Austria." Cultural Sociology 12, no. 2 (May 2, 2017): 193–207. http://dx.doi.org/10.1177/1749975517694299.
Abstract:
This article sets out to broaden our understanding of the significance of authenticity, locality and language for the development of a do-it-yourself (DIY) rap music career by taking male rap artists in Austria as an example. Drawing on interviews carried out in 2014–2015 with two groups of rap artists from different social and cultural backgrounds who embarked on their rap music careers in the early 1990s and the early 2000s, we analyse their rap lyrics and the social and economic contexts in which these individuals became rappers. We examine how the artists articulate claims to authenticity by appropriating African-American rap styles, meanings and idioms and blending them with local languages and references to local cultures and national politics. We also examine the rappers’ relationship to the music industry and the use of informal channels for the production, performance and consumption of rap and hip hop in general. The article suggests that the DIY careers of these rap artists depend on both the rappers’ use of music to articulate claims to authenticity and their ability to form (trans-)local networks for sharing skills, knowledge and other resources, as well as on Austria’s cultural policy and the changes in the music industry that have taken place in recent years.
22
Muh Kamim, Anggalih Bayu. "Pembangunan dan Ekonomi Politik Turisme Massal di Daerah Istimewa Yogyakarta." Jurnal Wacana Kinerja: Kajian Praktis-Akademis Kinerja dan Administrasi Pelayanan Publik 26, no. 1 (June 27, 2023): 51. http://dx.doi.org/10.31845/jwk.v26i1.791.
Abstract:
This research aimed to explore the transition from community-based tourism into mass tourism which has been initiated by the government. Data was collected through in depth interviews, observation, and secondary data analysis. This research found that the central government has been creating mass tourism since the ‘80s, because the state was interested in making tourism become one of the main incomes after the end of oil boom era. Mass tourism was booming until the end of the ‘90s. Nevertheless, the decrease in international tourist arrival was occurred because of forest fires, economic crises, Bali Bombing I and II, and SARS pandemic. Unfortunately, Yogyakarta’s earthquake in 2006 has hindered the recovery process. One decade after the earthquake, a mass tourism was booming again which was marked by the expansion of hotel construction. There were negative impacts such as lack of water access for public, repression of the labour movement, and policies that neglected public participation. The government continued to develop mass tourism through RPJMD DIY 2017-2022 as the nexus of central and local government interests. Projects from RPJMD DIY 2017-2022 were done by cultures and repressions mobilization, thus lead to another problems such as worsened public interests marginalization, land and labour privatization.
23
Stanley, Phiona. "Crafting a DIY Campervan and Crafting Embodied, Gendered Identity Performances in a Hyper-masculine Environment." Art/Research International: A Transdisciplinary Journal 4, no. 1 (February 27, 2019): 351–80. http://dx.doi.org/10.18432/ari29382.
Abstract:
This paper presents a multi-media textual collage that shows rather than tells the lived experiences of my conversion of a DIY campervan over several months in a diesel mechanic workshop in Sydney, Australia. This is a “small culture,” (Holliday, 1999) to which I gained limited access as I developed craft skills and the confidence to speak back to relative, milieu-specific, gendered power. I use autoethnographic textual fragments written shortly after the moment to depict the struggle to acquire skills, build confidence, and cross “small” cultures in an unusual crafting context. Grounded theoretical insights are suggested as they relate to three things. First, I examine the nature of individual, self-directed learning as engendered by the non-expert, hands-on doing of craft supported by YouTube instructional videos. Second, I consider positive and negative affective identity factors, particularly feelings of competence or incompetence and challenges to my own (female, middle-aged, injured, and non-expert) embodiment. Third, I consider the collaborative, discursive ways in which hegemonic and non-hegemonic masculinities were talked into being as contingent, relational identities against the foil of a constructed “other.”
24
Afriliani, Dhuwik, and Setiati Widihastuti. "Batak Toba’s Diaspor Loyalty against Customary Heritage in Special Region of Yogyakarta." SOCIA: Jurnal Ilmu-Ilmu Sosial 19, no. 2 (February 9, 2023): 19–31. http://dx.doi.org/10.21831/socia.v19i2.58456.
Abstract:
Various problems due to meetings between two or more cultures such as shock culture, cultural assimilation, and cross culture are considered to be problems of the diaspor community until now. The question is, is this problem also experienced by the Batak Toba’s diaspor, which incidentally is considered a tribe that is very loyal to the customs of their ancestral heritage? Then, how their efforts to maintain the sustainability of traditional marriages? To answer this question, a study was conducted on 8 (eight) Batak Toba’s diaspor in Special Region of Yogyakarta (DIY). The main findings in the study raised from descriptive research with a qualitative approach include: love of the Batak’s custom and the existence of social sanctions encourage the Batak Toba’s diaspor to hold tight to the tradition up to now, it encourages them to try to instill customary valuesand principles, provide direct practical learning to naposo, held traditional seminars, persuaded to directly deal with each marriage, and actively involved in their respective pockets so that their traditional marriages would be preserved. However, in this effort there are still some obstacles, one of which is the indifference of naposo in the management of the HKBP Church. This study recommends the need to allocate a portion of HKBP Church funds to provide guidance by providing free housing for naposo who migrate to DIY.
25
Pujianto, Ahmad. "Penetapan Gubernur dan Wakil Gubernur Daerah Istimewa Yogyakarta Perspektif Fikih Siyâsah." al-Daulah: Jurnal Hukum dan Perundangan Islam 5, no. 1 (April 1, 2015): 258–85. http://dx.doi.org/10.15642/ad.2015.5.1.258-285.
Abstract:
Abstract: This article discusses about the Islamic political jurisprudence perspective against the determination of the Governor and Deputy Governor of Yogyakarta. Law No. 13 year 2012 on specialty of Yogyakarta as a Special Region provides a constitutional basis in establishing that the Governor and Deputy Governor of Yogyakarta was enthroned as Sultan and to the Deputy Governor as Adipati Paku Alam. However, in the case of succession of the Governor of Yogyakarta, there is an exception because it is basically a privilage of DIY that has been existing since Indonesia had not gotten its independent yet and it is also guaranteed by the constitution. The direct determination of Sultan and Paku Alam is intended to award to the diversity of customs and cultures among the people of different regions. It is so because the determination of the Governor and Deputy Governor of Yogyakarta should directly be addressed wisely by all circles.Keywords: Governor, Yogyakarta, Islamic political jurisprudence.
26
Fuchsberger, Verena, Martin Murer, Manfred Tscheligi, Silvia Lindtner, Andreas Reiter, Shaowen Bardzell, Jeffrey Bardzell, and Pernille Bjørn. "The Future of Making: Where Industrial and Personal Fabrication Meet." Aarhus Series on Human Centered Computing 1, no. 1 (October 5, 2015): 4. http://dx.doi.org/10.7146/aahcc.v1i1.21394.
Abstract:
<div class="page" title="Page 1"><div class="layoutArea"><div class="column"><p><span>This one-day workshop seeks to reflect on the notion of fab- rication in both personal and industrial contexts. Although these contexts are very distinct in their economical and polit- ical vision, they share important characteristics (e.g., users interacting with specific fabrication equipment and tools). The workshop topic spans from personal fabrication to (au- tomated) production, from applied to theoretical considera- tions, from user requirements to design as a crafting practice. We will address changes in production that affect humans, e.g., from mass production to Do-It-Yourself (DIY) produc- tion, in order to discuss findings and lessons learned for in- dividual and collective production workplaces of the future. We aim to explore the intersections between different dimen- sions and processes of production ranging all the way from hobbyist to professional making. Furthermore, the workshop will critically reflect on current developments and their conse- quences on personal, societal, and economical levels includ- ing questions of the reorganization of work and labor, inno- vation cultures, and politics of participation. </span></p></div></div></div>
27
A, Eisenberg. "How did the Cultural Revolution affect your Culture?" Journal of Natural & Ayurvedic Medicine 4, no. 3 (July 6, 2020): 1–4. http://dx.doi.org/10.23880/jonam-16000270.
Abstract:
While serving as International Expert at the Research Institute of Anthropology and Ethnology, Jishou University in Xiangxi Autonomous Prefecture of Hunan Province, China, on United Nations Educational, Scientific and Cultural Organization-Local and Indigenous Knowledge Systems (UNESCO-LINKS) Natural Science Sector, United Nations Permanent Forum on Indigenous Issues (UNPFII), United Nations Department of Economic and Social Affairs (UNDESA) projects with the Kam people of China and ministries responsible for ethnic development, I asked my ethnic minority graduate students and colleagues of China this question, “How did the Cultural Revolution affect your culture?”, and they thoughtfully shared their perspectives and experiences.
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Poei, Eliza Purnamasari, and J. Dwijoko Ansusanto. "PERILAKU BERLALU LINTAS YANG MENDUKUNG KESELAMATAN DI JALAN RAYA." Jurnal Teknik Sipil 14, no. 1 (April 5, 2017): 10. http://dx.doi.org/10.24002/jts.v14i1.1012.
Abstract:
Yogyakarta as student city and tourist destination that will cause the number of immigrantsstudying or sightseeing in Special Region of Yogyakarta increasing from year to year. Yogyakartais like a miniature Indonesia, a wide variety of people with cultures and tribes can befound here, they perform daily activities with their behavior and habits of each. The number ofmotorvehicles in Yogyakarta also increased from year to year. Motor vehicle technology is moreadvanced offset by road users catch up with the times of fast-paced, causing traffic on the highwayis getting crowded. If road users is not an orderly way, behaving arbitrarily can endanger otherroad users. The purpose of this study are : a) To investigate the behavior of road users, both driversand pedestrians in DIY, b) analyzing public opinion on the behavior of road users when passtrafficon the highway. Direct observation in the field and deployment questionnaire conducted inDIY. The analysis showed that most motorcyclists do not obey traffic sign or jumping red. Overtakethe vehicle from the left side of the overtaken vehicle. Turn right or left is not lit the lampsign. That respondents believe that the implementation of orderly traffic both closely associatedwith “The application of sanctions/penalties consequently” (64%). Accident ever experienced 54%of respondents in the province is a collision with a motorcycle. The main cause of accidents due tolack of concentration, 33% of accidents occurred at noon. According to respondents the reasondoes not obey traffic signs / road markings 65% because “ there are no policemen watching”.Usually respondents APILL light violation 61% at the moment “in a hurry because nearly/toolate to school/work/point of interest. Abstrak : Yogyakarta sebagai kota pelajar dan daerah tujuan Wisata menyebabkan Jumlah pendatangyang akan menuntut ilmu maupun berwisata di Daerah Istimewa Yogyakarta semakinmeningkat dari tahun ke tahun, Yogyakarta bagaikan miniaturnya Indonesia, berbagai ragam orangdengan budaya dan suku dapat dijumpai disini, mereka melakukan aktifitas sehari-hari dengan perilakudan kebiasaan mereka masing-masing. Jumlah kendaraan bermotor yang ada di DIY jugameningkat dari tahun ke tahun. Teknologi kendaraan bermotor yang semakin maju diimbangi olehpengguna jalan yang mengikuti derap langkah perkembangan zaman yang serba cepat, menyebabkanlalu-lintas di jalan raya semakin padat. Jika pengguna jalan tidak tertib, berperilaku semaunyasendiri dapat membahayakan keselamatan pengguna jalan yang lain.. Tujuan penelitian iniadalah : a) Menginvestigasi perilaku pengguna jalan , baik pengemudi maupun penyeberang jalandi DIY, b) Menganalisis penilaian masyarakat terhadap perilaku pengguna jalan saat berlalu-lintasdi jalan raya. Pengamatan langsung di lapangan dan penyebaran questioner dilakukan di wilayahDIY.Hasil analisis menunjukkan bahwa sebagian besar pengguna sepeda motor tidak mentaatirambu maupun APILL, menyalip kendaraan dari sisi kiri kendaraan yang disalip, berbelok kananatau kiri tidak memberi tanda lampu sign . Pendapat responden bahwa pelaksanaan tertib lalulintasyang baik berkaitan erat dengan “Penerapan sanksi/hukuman secara konsekuen”(64%). Kecelakaanyang pernah dialami responden di DIY 54% adalah tabrakan dengan sepeda motor, sebabutama kecelakaan karena kurang konsentrasi (38%), kecelakaan terjadi 33% pada siang hari..Menurut responden alasan tidak mematuhi rambu lalu-lintas /marka jalan 65% karena “tidak adapolisi yang mengawasi”. Biasanya responden melanggar lampu APILL pada saat “Tergesa-gesakarena hampir /sudah terlambat ke sekolah/tempat kerja/tempat tujuan” (61%).Kata kunci:: lalu-lintas, perilaku, keselamatan, pengguna jalan, kendaraan
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Tsouroufli, Maria. "Gendered and classed performances of ‘good’ mother and academic in Greece." European Journal of Women's Studies 27, no. 1 (September 24, 2018): 9–24. http://dx.doi.org/10.1177/1350506818802454.
Abstract:
The enduring significance of gender and how it intersects with class in the organization of parenting, domestic and professional work has been obscured in contemporary neoliberal contexts. This article examines how Greek academic women conceptualize and enact motherhood and the classed and gendered strategies they adopt to reconcile ‘good’ motherhood with notions of the ‘good’ academic professional. It draws on semi-structured interviews about the career narratives of 15 women in Greek medical schools in the aftermath of the Greek recession. The analysis presented in this article is informed by a feminist post-structuralist paradigm and an emic approach to intersectionality. Motherhood emerged in the data as a dynamic concept, and a network of practices both constrained and enabled by gendered and classed family and work cultures. Drawing on a neoliberal ‘DIY’ and ‘having it all’ discourse, Greek mothers claimed that they could achieve almost anything professionally, if they organized their private lives sensibly. They drew on idealized discourses of motherhood, but they also contradicted these notions by doing non-traditional forms of motherhood, such as remote or transnational motherhood, afforded by their privileged social positioning and academic careers. Further research is required to investigate configurations of classed motherhood in less prestigious professions.
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Fabelo, Dora M., Krish Stella, Natascha Barreto-Romero, and Nancy Valdez-Gainer. "Professional Book Reviews: The Rights of Readers in Our Schools." Language Arts 88, no. 3 (January 1, 2011): 238–43. http://dx.doi.org/10.58680/la201113418.
Abstract:
Dora M. Fabelo, Krish Stella, Natascha Barreto-Romero, Nancy Valdez-GainerFour professionals who represent different capacities in one urban elementary school—principal, reading specialist, librarian, and classroom teacher—each select one book to review relating to the theme of “rights of readers.” The first book reviewed, A Declaration of Reader’s Rights: Renewing Our Commitment to Students, written by Bass, Dasinger, Elish-Piper, Matthews, and Risko, provides an overview of what is meant by “rights of readers” and what can be done in schools to address this issue. Next, Burn This Book: PEN Writers Speak Out on the Power of the Word, edited by Toni Morrison, contains diverse chapters relating how censorship infringes on the rights of readers. DIY Media: Creating, Sharing, and Learning with New Technologies, edited by Knobel and Lankshear, examines do-it-yourself media as social literacy practice, thus calling for the rights of readers to engage with expanded definitions of what should count as text. Finally, Teaching for Joy and Justice: Re-Imagining the Language Arts Classroom, written by Linda Christensen, highlights the possibilities that arise when students are given the right to engage with reading and writing that reflects their diverse lives, cultures, and languages, affording opportunities to read and write social justice texts.
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Goodley, Dan, David Cameron, Kirsty Liddiard, Becky Parry, Katherine Runswick-Cole, Ben Whitburn, and Meng Ee Wong. "Rebooting Inclusive Education? New Technologies and Disabled People." Canadian Journal of Disability Studies 9, no. 5 (December 20, 2020): 515–49. http://dx.doi.org/10.15353/cjds.v9i5.707.
Abstract:
This paper provides a speculative, conceptual and literature-based review of the relationship between disability and new technologies with a specific focus on inclusive education for disabled people. The first section critically explores disability and new technologies in a time of Industry 4.0. We lay out some concerns that we have, especially in relation to disabled people’s peripheral positionality, when it comes to these new developments. The second section focuses on the area of inclusive education. Inclusion and education are oftentimes in conflict with one another. We tease out these conflicts and argue that we cannot decouple the promise of new technologies from the challenges of inclusive education, because, in spite of the potential for technological mediation to broaden access to education, there remains deep-rooted problems with exclusion. The third section of our paper explores affirmative possibilities in relation to the interactions between disability and new technologies. We draw on the theoretical fields of Science and Technology Studies; Critical Disability Studies; Assistive and Inclusive Technologies; Collaborative Robotics, Maker and DIY Cultures and identify a number of key considerations that relate directly to the revaluing of inclusive education. We conclude our paper by identifying what we view as pressing and immediate concerns for inclusive educators when considering the merging of disability and technology, accessibility and learning design.
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Pechlivani, Eleftheria Maria, Sotirios Pemas, Alexandros Kanlis, Paraskevi Pechlivani, Spyros Petrakis, Athanasios Papadimitriou, Dimitrios Tzovaras, and Konstantinos E. Hatzistergos. "Enhanced Growth of Bacterial Cells in a Smart 3D Printed Bioreactor." Micromachines 14, no. 10 (September 26, 2023): 1829. http://dx.doi.org/10.3390/mi14101829.
Abstract:
In the last decade, there has been a notable advancement in diverse bioreactor types catering to various applications. However, conventional bioreactors often exhibit bulkiness and high costs, making them less accessible to many researchers and laboratory facilities. In light of these challenges, this article aims to introduce and evaluate the development of a do-it-yourself (DIY) 3D printed smart bioreactor, offering a cost-effective and user-friendly solution for the proliferation of various bioentities, including bacteria and human organoids, among others. The customized bioreactor was fabricated under an ergonomic design and assembled with 3D printed mechanical parts combined with electronic components, under 3D printed housing. The 3D printed parts were designed using SOLIDWORKS® CAD Software (2022 SP2.0 Professional version) and fabricated via the fused filament fabrication (FFF) technique. All parts were 3D printed with acrylonitrile butadiene styrene (ABS) in order for the bioreactor to be used under sterile conditions. The printed low-cost bioreactor integrates Internet-of-things (IoT) functionalities, since it provides the operator with the ability to change its operational parameters (sampling frequency, rotor speed, and duty cycle) remotely, via a user-friendly developed mobile application and to save the user history locally on the device. Using this bioreactor, which is adjusted to a standard commercial 12-well plate, proof of concept of a successful operation of the bioreactor during a 2-day culture of Escherichia coli bacteria (Mach1 strain) is presented. This study paves the way for more in-depth investigation of bacterial and various biological-entity growth cultures, utilizing 3D printing technology to create customized low-cost bioreactors.
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Migliaccio, Giovanni, Francesca Masiello, Valentina Tirelli, Massimo Sanchez, Lilian Varricchio, Carolyn Whitsett, and Anna Rita F. Migliaccio. "Mature Erythroblasts Die of Autophagy and Induce Autophagy of Immature Erythroblasts, Limiting the Final Cellular Output In Human Erythroid Massive Amplification (HEMA) Culture." Blood 116, no. 21 (November 19, 2010): 339. http://dx.doi.org/10.1182/blood.v116.21.339.339.
Abstract:
Abstract Abstract 339 Mononuclear cells from adult blood (buffy coat) generate great numbers (∼1010) of erythroid cells (EBs) in culture stimulated with stem cell factor (SCF), interleukin-3 (IL-3), erythropoietin (EPO), dexamethasone and estrogens (human erythroid massive amplification, HEMA) but the number is below that required for one transfusion (∼1012) highlighting the importance of devising strategies to further improve cell proliferation ex-vivo. From day 6 to day 21, two types of EBs are recognized using CD36 (the thrombospondin receptor) and CD235a (Glycophorin A) expression: immature CD36highCD235aneg EBs (iEBs) and mature CD36highCD235ahigh EBs (mEBs). The aim of this study was to identify the contribution of iEBs and mEBs to the final EBs cellular output in HEMA. Cell isolation and functional studies indicated that iEBs do not grow in semisolid media but are able to generate additional EBs when cultured in HEMA (FI after 48 hrs=2.57±1.15). By contrast, mEBs die within 4–5 days. In addition, iEBs sequentially sorted every 2 days generate 5-fold more EBs than unfractionated cells cultured in parallel, suggesting that mEBs may inhibit growth of iEBs. To test this hypothesis, day 10 iEBs and mEBs separated by sorting were co-cultured for 48 hrs at ratios of 100/0, 5/1, 5/3 and 1/2 for fate determination (proliferation, maturation and/or death). Either iEBs or mEBs were labelled by CSFE staining to account for possible toxic effects of CSFE-labeling. iEBs cultured alone increased in numbers by 2-fold over 4 days whereas iEBs when co-cultured with mEB did not generate additional cells (5/1-5/3 ratios) and even died, becoming undetectable (1/2 ratios). Although cell numbers did not increase in co-cultures, proliferation indexes (1.3, based on hemi-decrements of CSFE labelling), maturation (50% of newly generated cells were mEBs) and apoptosis (barely detectable Annexin Vpos cells) were similar when iEBs were cultured alone or in combination with mEBs. These observations suggest that failure to observe increases in cell numbers in the co-cultures may be due to non-apoptotic cell death, possibly autophagy. EBs activate autophagy as part of the maturation process by forming the autophagic vescicles required for organelle desctruction. One of the first steps in the formation of autophagic vescicles is the conversion by lipidation of the cytosolic form of the microtubule associated protein light chain 3 (LC3-I) into the vescicle-specific LC3-II form. Once the autophagic vescicles are prepared, EBs either progress to enucleation or die by autophagy. Acridine Orange (AO) staining may be used to monitor authophagic death. By western blot analyses, day 10–11 EBs express LC3-I/LC3-II ratios of 1:2, indicating that the autophagic machinery is already maximally activated in these cells. By FACS analyses, however, only a fraction (3-4%) of day 10–11 EBs are stained by AO. However, these cells are susceptible to induction of autophagic death because growth factor starvation (STV) for 4 hrs does not induce apoptosis (Annexin Vpos cells: 6.6±4.8% vs 10.7±7.3% before and after STV, respectively) or change LC3-I/LC3-II ratio (which remains 1:2) but greatly increases the AOpos cells (4.3% to 23.1±3.7%, p<0.01). To clarify the role of autophagy in limiting the cell output in HEMA, the number of AOpos cells observed in day 10 iEBs and mEBs cultured for 4 days either alone or at the 1:1 ratio was determined. Increases in total cell numbers were observed in cultures of iEBs alone (FI=4) but not in those of mEBs, either alone or with iEBs (FI=1 in both). AOpos cells were barely detectable in cultures of iEBs alone (1.6%) but became 23% and 17% in those of mEBs alone and with iEBs, respectively. Calculations based on numbers of mEBs seeded in cultures (3×105 alone and 1.5×105 with 1.5×105 iEBs) and on FI observed (1) indicate that the number of AOpos cells is 7×104 (23% of 3×105 cells) when mEBs are cultured alone and 5.1×104 (17% of 3×105 cells) in iEBs/mEBS coculture, instead of 3.5×104 (23% of 1.5×105 cells) AOpos cells expected to be generated by the mEBs added to the co-culture. This difference suggests that during co-culture, mEBs induce iEBs to die by autophagy. In conclusion, these data indicate that the autophagic pathway is maximally activated in day 10 EBs and plays a major role in limiting the final cellular output of HEMA. Disclosures: No relevant conflicts of interest to declare.
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Becker, S., M. K. Warren, and S. Haskill. "Colony-stimulating factor-induced monocyte survival and differentiation into macrophages in serum-free cultures." Journal of Immunology 139, no. 11 (December 1, 1987): 3703–9. http://dx.doi.org/10.4049/jimmunol.139.11.3703.
Abstract:
Abstract The role of mononuclear phagocyte-specific colony-stimulating factor (CSF-1) in human monocyte to macrophage differentiation was investigated. The addition of 1000 U/ml of CSF-1 to serum-free monocyte cultures resulted in monocyte survival comparable to that in cultures containing 5% AB serum, whereas cells in serum- and CSF-1-free medium lost their viability in 3 to 5 days. The requirement for CSF-1 coincided with the time (40 to 64 hr of culture) when the major changes in morphology and biochemical function took place in monocytes undergoing differentiation into macrophages. If CSF-1 was removed from the cultures before this time, death of the monocytes resulted. In cultures containing CSF-1, as in serum containing cultures, the lysosomal enzyme acid phosphatase was enhanced 10- to 20-fold by day 4 to 5. Superoxide production in response to phorbol myristic acetate was maintained in CSF-1 cultured monocytes, but declined with time in monocytes cultured in serum. The expression of monocyte-macrophage antigens p150.95 (LeuM5), OKM1, LeuM3, Fc receptors (32.2), and HLA-DR had increased in CSF-1 containing cultures at day 4. When antigen expression was analyzed at day 2 to 3, when cell size and 90 degrees scatter characteristics were still identical to control serum-free cultures, only p150.95, HLA-DR and FcR expression were enhanced by CSF-1. Low amounts of lipopolysaccharide (0.1 ng/ml) were found to enhance monocyte survival in the absence of added CSF-1. Lipopolysaccharide-containing cultures were found to produce CSF-1 (up to 450 U/ml, as detected by radioimmunoassay). Lipopolysaccharide (1 microgram/ml), however, did not induce enhanced expression of the maturation-related antigens. Based on these observations we conclude that CSF-1 is enhancing human monocyte survival and is involved in the events leading to the differentiation of monocytes into macrophages.
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Purton, Louise E., Irwin D. Bernstein, and Steven J. Collins. "All-Trans Retinoic Acid Delays the Differentiation of Primitive Hematopoietic Precursors (lin−c-kit+Sca-1+) While Enhancing the Terminal Maturation of Committed Granulocyte/Monocyte Progenitors." Blood 94, no. 2 (July 15, 1999): 483–95. http://dx.doi.org/10.1182/blood.v94.2.483.
Abstract:
Abstract All-trans retinoic acid (ATRA) is a potent inducer of terminal differentiation of malignant promyelocytes, but its effects on more primitive hematopoietic progenitors and stem cells are less clear. In this study, we investigated the effect of ATRA on highly enriched murine hematopoietic precursor cells (lin−c-kit+Sca-1+) grown in liquid suspension culture for 28 days. ATRA initially slowed the growth of these hematopoietic precursors but prolonged and markedly enhanced their colony-forming cell production compared with the hematopoietic precursors cultured in its absence. At 7 and 14 days of culture, a substantially greater percentage of cells cultured with ATRA did not express lineage-associated antigens (55.4% at day 7 and 68.6% at day 14) and retained expression of Sca-1 (44.7% at day 7 and 79.9% at day 14) compared with cells grown in its absence (lin−cells: 31.5% at day 7 and 4% at day 14; Sca-1+: 10.4% at day 7 and 0.7% at day 14). Moreover, a marked inhibition of granulocyte production was observed in cultures continuously incubated with ATRA. Significantly, ATRA markedly prolonged and enhanced the production of transplantable colony-forming unit-spleen (CFU-S) during 14 days of liquid suspension culture. In contrast with its effects on primitive lin−c-kit+Sca-1+hematopoietic precursors, ATRA did not exert the same effects on the more committed lin−c-kit+Sca-1−progenitor cells. Moreover, the late addition of ATRA (7 days post-culture initiation) to cultures of primitive hematopoietic precursors resulted in a marked decrease in colony-forming cell production in these cultures, which was associated with enhanced granulocyte differentiation. These observations indicate that ATRA has different effects on hematopoietic cells depending on their maturational state, preventing and/or delaying the differentiation of primitive hematopoietic precursors while enhancing the terminal differentiation of committed granulocyte/monocyte progenitors.
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Purton, Louise E., Irwin D. Bernstein, and Steven J. Collins. "All-Trans Retinoic Acid Delays the Differentiation of Primitive Hematopoietic Precursors (lin−c-kit+Sca-1+) While Enhancing the Terminal Maturation of Committed Granulocyte/Monocyte Progenitors." Blood 94, no. 2 (July 15, 1999): 483–95. http://dx.doi.org/10.1182/blood.v94.2.483.414k12_483_495.
Abstract:
All-trans retinoic acid (ATRA) is a potent inducer of terminal differentiation of malignant promyelocytes, but its effects on more primitive hematopoietic progenitors and stem cells are less clear. In this study, we investigated the effect of ATRA on highly enriched murine hematopoietic precursor cells (lin−c-kit+Sca-1+) grown in liquid suspension culture for 28 days. ATRA initially slowed the growth of these hematopoietic precursors but prolonged and markedly enhanced their colony-forming cell production compared with the hematopoietic precursors cultured in its absence. At 7 and 14 days of culture, a substantially greater percentage of cells cultured with ATRA did not express lineage-associated antigens (55.4% at day 7 and 68.6% at day 14) and retained expression of Sca-1 (44.7% at day 7 and 79.9% at day 14) compared with cells grown in its absence (lin−cells: 31.5% at day 7 and 4% at day 14; Sca-1+: 10.4% at day 7 and 0.7% at day 14). Moreover, a marked inhibition of granulocyte production was observed in cultures continuously incubated with ATRA. Significantly, ATRA markedly prolonged and enhanced the production of transplantable colony-forming unit-spleen (CFU-S) during 14 days of liquid suspension culture. In contrast with its effects on primitive lin−c-kit+Sca-1+hematopoietic precursors, ATRA did not exert the same effects on the more committed lin−c-kit+Sca-1−progenitor cells. Moreover, the late addition of ATRA (7 days post-culture initiation) to cultures of primitive hematopoietic precursors resulted in a marked decrease in colony-forming cell production in these cultures, which was associated with enhanced granulocyte differentiation. These observations indicate that ATRA has different effects on hematopoietic cells depending on their maturational state, preventing and/or delaying the differentiation of primitive hematopoietic precursors while enhancing the terminal differentiation of committed granulocyte/monocyte progenitors.
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YAN, RUN-TAO, and SHU-ZHEN WANG. "Differential induction of gene expression by basic fibroblast growth factor and neuroD in cultured retinal pigment epithelial cells." Visual Neuroscience 17, no. 2 (March 2000): 157–64. http://dx.doi.org/10.1017/s0952523800171172.
Abstract:
Embryonic chick retinal pigment epithelial (RPE) cells can undergo transdifferentiation upon appropriate stimulation. For example, basic fibroblast growth factor (bFGF) induces intact RPE tissue younger than embryonic day 4.5 (E4.5) to transdifferentiate into a neural retina. NeuroD, a gene encoding a basic helix-loop–helix transcription factor, triggers de novo production of cells that resemble young photoreceptor cells morphologically and express general neuron markers (HNK-1/N-CAM and MAP2) and a photoreceptor-specific marker (visinin) from cell cultures of dissociated E6 RPE (Yan & Wang, 1998). The present study examined whether bFGF will lead to the same transdifferentiation phenomenon as neuroD when applied to dissociated, cultured E6 RPE cells, and whether interplay exists between the two factors under the culture conditions. Dissociated E6 RPE cells were cultured in the presence or absence of bFGF, and with or without the addition of retrovirus expressing neuroD. Gene expression was analyzed with immunocytochemistry and in situ hybridization. Unlike neuroD, bFGF did not induce the expression of visinin, or HNK-1/N-CAM and MAP2. However, bFGF elicited the expression of RA4 immunogenicity; yet, many of these RA4-positive cells lacked a neuronal morphology. Addition of bFGF to neuroD-expressing cultures did not alter the number of visinin-expressing cells; misexpression of neuroD in bFGF-treated cultures did not change the number of RA4-positive cells, suggesting the absence of interference or synergistic interaction between the two factors. Our data indicated that bFGF and neuroD induced the expression of different genes in cultured RPE cells.
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Lackner, Andreas, Heinz Stammberger, Walter Buzina, Kurt Freudenschuss, Thomas Panzitt, Stefan Schosteritsch, and Hannes Braun. "Fungi: A Normal Content of Human Nasal Mucus." American Journal of Rhinology 19, no. 2 (March 2005): 125–29. http://dx.doi.org/10.1177/194589240501900203.
Abstract:
Background In recent studies, we showed that 91.3% of both CRS patients and healthy controls grew positive fungal cultures out of their nasal mucus, which therefore appears to be a common finding within the adult population. However, it still was unknown as of when fungi could be cultured from nasal mucus in human beings. We attempted to ascertain this point of time in the nasal mucus of neonates. Methods We examined nasal mucus from 30 neonates immediately after birth, on the 1st and 4th day postpartum and after 2 and 4 months of life. The samples obtained with sterile cotton swabs were cultured on agar plates. Fungal cultures were identified either conventionally by microscopy or with molecular techniques. To prove possible contamination during birth, mucus of the maternal birth canal was examined as well. Results In 6 of 30 (20%) of our neonates we found positive fungal cultures immediately after birth in (3 of them Candida albicans) most likely because of contamination passing the maternal birth canal. In 2 of 29 (7%) of our neonates, positive fungal cultures were obtained on the 1st day postpartum, and in 4 of 26 (15%) positive fungal cultures were obtained on the 4th day, all limited to 1 day only and without clinical symptoms of colonization. After the 2nd month of life, examination of nasal mucus yielded positive fungal cultures in 8 of 11 (72%), and after 4 months examination of nasal mucus yielded positive fungal cultures in 17 of 18 (94%) of our babies, with a wide array of different species. Conclusions Fungi can be cultured from nasal mucus as soon as contact with the environmental air exists but they are not persistent in the 1st day of life. However, after 4 months, the situation is similar to the one in adults: fungal cultures can be obtained from almost everyone's nose. Therefore, fungi must be considered a normal content of nasal mucus.
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Barnard, R., G. Thordarson, M. F. Lopez, M. Yamaguchi, A. Edens, S. D. Cramer, L. Ogren, and F. Talamantes. "Expression of growth hormone-binding protein with a hydrophilic carboxyl terminus by the mouse placenta: studies in vivo and in vitro." Journal of Endocrinology 140, no. 1 (January 1994): 125–35. http://dx.doi.org/10.1677/joe.0.1400125.
Abstract:
Abstract GH-binding protein (GHBP) or GH receptor is present in numerous extrahepatic tissues in the rodent. From mid- to late gestation in the mouse, the maternal serum concentration of GHBP increases 30- to 50-fold. We have investigated whether the placenta might synthesize GHBP and potentially contribute to this increase. RNA was isolated from placentas and subjected to Northern analysis using a cDNA probe to the shared region of GHBP and GH receptor-encoding mRNAs. From day 8 to day 18 of gestation, the GHBP-encoding mRNA (1·4 kb) increased 45-fold in quantity. The GH receptor-encoding mRNA (4·2 kb) increased sixfold by day 14 and then remained steady until day 18. These changes which were not co-ordinated parallel reported changes in the steady-state concentrations of 1·4 and 4·2 kb mRNAs in maternal liver, suggesting shared regulatory factors. Extracts of freshly isolated trophoblasts were assayed for GHBP with a radioimmunoassay specific for GHBP with a hydrophilic carboxyl terminus. The cytosolic content of immunoreactive GHBP increased fourfold from mid- to late gestation. Trophoblasts were isolated from placentas and cultured for 2 days on collagen gels in defined medium. Cultured cells were at least 90% viable and secreted mouse placental lactogen-II (mPL-II). Immunocytochemistry was carried out simultaneously on cells cultured from day 7 to day 17 of gestation using a monoclonal antibody (MAb 4·3), which recognizes the hydrophilic C-terminus of GHBP. Cell-localized GHBP was present in trophoblasts cultured for 2 days, but GHBP was not detectable by radioimmunoassay or by immunoprecipitation in concentrated culture media from cultures treated with 100 ng mouse GH/ml or 100 ng mPL-II/ml or from untreated cultures. RNA was isolated from cells cultured in an identical manner to those analysed by immunocytochemistry. Three GH receptor/GHBP mRNA species of 8, 4·2 and 1·4 kb were observed. The quantity of 4·2 and 1·4 kb mRNAs did not change significantly in cultures from day 7 to day 15 of gestation but, in cultures from day 17 of gestation, the amount of 1·4 kb mRNA dropped significantly, while that of the 4·2 kb mRNA remained unchanged. GHBP- and GH receptor-encoding mRNAs are not co-ordinately regulated in vivo or in vitro. Although mPL-II was secreted into the medium by cultured trophoblasts, secretion of GHBP could not be detected. The culture medium may not contain the specific factors required for secretion of placental GHBP, or placental GHBP may not be destined for secretion. The results show that GHBP (as distinct from GH receptor) is expressed by the placenta in vivo and trophoblasts in vitro. From mid-gestation onwards, GHBP mRNA increases dramatically in vivo and the cytosolic content of GHBP in freshly isolated trophoblasts increases. This suggests an important local regulatory role for placental GHBP during gestation. Journal of Endocrinology (1994) 140, 125–135
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Arambepola, NK, D. Bunick, and PS Cooke. "Thyroid hormone effects on androgen receptor messenger RNA expression in rat Sertoli and peritubular cells." Journal of Endocrinology 156, no. 1 (January 1, 1998): 43–50. http://dx.doi.org/10.1677/joe.0.1560043.
Abstract:
Postnatal Sertoli cell maturation is characterized by a pronounced rise in androgen receptor (AR) expression, which increases several fold between birth and adulthood. Since both 3,3',5-triiodothyronine (T3) and FSH regulate Sertoli cell proliferation and differentiation, we have determined the effects of T3 and FSH on AR mRNA expression in cultured Sertoli cells from 5-day-old rats. These cultures contain 5-9% peritubular cells, which also express AR mRNA. To insure that the observed T3 responses did not result from peritubular cells, we examined T3 effects on AR mRNA expression in cultured 20-day-old Sertoli cells (which contain minimal peritubular contamination) and peritubular cells, and measured thyroid hormone receptor (TR) mRNA expression in both of these cell types. Sertoli cells from 5- and 20-day-old rat testes were grown in serum-free medium alone (controls) or with ovine FSH (100 ng/ml) and/or T3 (100 nM) for 4 days. Peritubular cells purified from 20-day-old rat testes were grown in serum-containing medium for 8 days. These cells were split 1:4, and grown an additional 8 days, the last 4 days in serum-free medium with or without T3. TR and AR mRNA levels in all cultures were determined by Northern blotting. AR mRNA levels in 5- and 20-day-old cultured Sertoli cells were significantly (P < 0.05) increased by both T3 and FSH alone. Furthermore, AR mRNA levels in Sertoli cells treated with T3 and FSH were greater than with either alone. TR mRNA expression was detected in cultured peritubular cells, but TR mRNA levels in these cells were only approximately 30% of that seen in 20-day-old cultured Sertoli cells. In contrast to Sertoli cells, T3 did not affect peritubular AR mRNA expression. These results indicate that T3 is an important regulator of the postnatal Sertoli cell AR mRNA increase. The additive effects of maximally stimulatory doses of FSH and T3 suggest these hormones work through different mechanisms to increase AR mRNA. TR mRNA expression in peritubular cells indicates these cells may be direct T3 targets, though the function of T3 in these cells is unknown.
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Huang, Xiaosong, L. Jeanne Pierce, Paul A. Cobine, Dennis R. Winge, and Gerald J. Spangrude. "Copper Modulates the Differentiation of Mouse Hematopoietic Progenitor Cells in Culture." Cell Transplantation 18, no. 8 (August 2009): 887–97. http://dx.doi.org/10.3727/096368909x471152.
Abstract:
Copper chelation has been shown to favor the expansion of human hematopoietic stem/progenitor cells in vitro. To further understand the effects of copper modulation on defined subsets of stem cells versus progenitor cells, we extended the studies in a mouse system. We isolated mouse hematopoietic stem cells (HSCs) or hematopoietic progenitor cells (HPCs) and cultured them with or without the copper chelator tetraethylenepentamine (TEPA) or CuCl2. Cytokine-stimulated HPC cultures treated with TEPA for 7 days generated about two to three times more total and erythroid colony-forming cells (CFCs) compared to control cultures. In contrast, CuCl2 treatment decreased the CFC numbers. Similar results were seen with HSC after 14, but not 7, days of culture. Transplant studies showed that HPCs cultured for 7 days in TEPA had about twofold higher short-term erythroid repopulation potential compared to control cultures, while CuCl2 decreased the erythroid potential of cultured HPCs compared to control cultures. HSCs cultured with TEPA for 7 days did not exhibit significantly higher repopulation potential in either leukocyte or erythrocyte lineages compared to control cultures in short-term or long-term assays. Based on JC-1 staining, the mitochondrial membrane potential of HPCs cultured with TEPA was lower relative to control cultures. Our data suggest that decreasing the cellular copper content with TEPA results in preferential expansion or maintenance of HPC that are biased for erythroid differentiation in vivo, but does not enhance the maintenance of HSC activity in culture.
42
Guo, W., K. Kamiya, J. Cheng, and J. Toyama. "Changes in action potentials and ion currents in long-term cultured neonatal rat ventricular cells." American Journal of Physiology-Cell Physiology 271, no. 1 (July 1, 1996): C93—C102. http://dx.doi.org/10.1152/ajpcell.1996.271.1.c93.
Abstract:
A primary culture of neonatal ventricular myocytes isolated from day-old rats was established for investigating the changes in action potentials and ion currents over long periods. Cells at days 5 and 15 in culture were studied. These changes in vitro were compared with those in situ derived from the age-matched freshly isolated cells. During primary culture, quiescent cells demonstrated shortening of action potential durations (APD) resembling the developmental changes observed in situ. The beating cultured cells were not associated with APD shortening. Despite constant current amplitudes, the densities of Ca2+ currents (ICa) decreased in the quiescent cultures at later ages as a result of cell enlargement. ICa densities were maintained in the beating cultured and freshly isolated cells. Acceleration in the inactivation of ICa was observed during developments both in vitro and in situ. In addition, the densities of transient outward currents (Ito) tripled and doubled in the quiescent and beating cells during 15-day cultures. However, Ito in beating cultured cells made less contribution to APD in contrast to the quiescent cultured and freshly isolated myocytes. These findings demonstrate that electrophysiological properties differ between two types of long-term cultured cells. ICa densities remained constant in the beating cultures, suggesting that cell beating may be required for the maintenance of ICa density in developing cardiomyocytes.
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Van Der Meulen, J., F. A. Helmond, G. T. Kronnie, and C. P. J. Oudenaarden. "The influence of endometrium on morphology and oestrogen release of day-11 and day-13 pig blastocysts in vitro." Netherlands Journal of Agricultural Science 42, no. 3 (September 1, 1994): 225–34. http://dx.doi.org/10.18174/njas.v42i3.600.
Abstract:
Day-11 spherical and Day-13 filamentous blastocysts were cultured in vitro without supplementation of oestrogen precursors for 2 consecutive periods of 24 h, with or without endometrial tissue. Spherical Day-11 blastocysts flushed from one uterine horn (5.5 +/- 0.8 blastocysts; 5.3 +/- 0.4 mm in diameter; mean +/- s.d.) did not show gross morphological changes during the culture period. Free and conjugated oestrone (E1) and oestradiol-17 beta (E2) were released during the first and second 24 h culture period and was maximal during the first 24 h. In the presence of endometrial tissue the release of oestrogens from the blastocysts was reduced to 30-50%. Clewed Day-13 filamentous blastocysts, recovered after flushing, formed trophospheres and also in these cultures both free and conjugated E1 and E2 were released. In this case the addition of endometrial tissue did not affect the release of oestrogens from the blastocysts. Single Day-13 filamentous blastocysts cultured without endometrial tissue (n = 33, from 4 gilts), developed trophospheres and all blastocysts released E1 and E2. During the first 24 h on average per blastocyst 2.6 ng E1 (range 1.6-4.0) and 9.7 ng E2 (range 4.3-15.3) were released. In all cultures of spherical and filamentous blastocysts E2 release exceeded E1 release.These data show that Day 11-13 blastocysts are able to release both free and conjugated E1 and E2 during in vitro culture from endogenous precursors. The addition of endometrial tissue reduced the release of oestrogens from Day-11 but not Day-13 blastocysts. Whether this is due to an altered oestrogen release of the blastocysts in the presence of endometrium on Day-11 or due, to an increased uptake of oestrogens by the endometrium on Day-11 remains to be investigated.
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Trøen, G., A. Nilsson, K. R. Norum, and R. Blomhoff. "Characterization of liver stellate cell retinyl ester storage." Biochemical Journal 300, no. 3 (June 15, 1994): 793–98. http://dx.doi.org/10.1042/bj3000793.
Abstract:
The stellate cells of the liver are the main storage site of retinyl esters in the body. During cultivation in vitro of stellate cells isolated from rat and rabbit livers were observed that the cells rapidly loose their retinyl ester content. Freshly isolated stellate cells contain about 144 nmol of total retinol/mg of protein, while cells cultivated for 14 days contained below 0.1 nmol/mg of protein. When 3-day-old cultures were incubated for 6 h with 2 microM retinol, the cellular content increased from 5.6 to approx. 9.4 nmol of total retinyl esters/mg of protein. In contrast, little retinyl ester accumulated in 10-20-day-old cultures incubated with 2 microM retinol. At 50 microM retinol, however, the retinyl ester level did increase both with 3-day-old cultures and 10-20-day-old cultures. In parallel experiments with cultured fibroblasts esterification characteristics similar to those seen in older cultures of stellate cells were observed. When 10-day-old cultures of stellate cells were incubated with retinol alone, or in combination with palmitic acid, linoleic acid or oleic acid, the total storage of retinyl esters increased by 20-150%. In most cases, the fatty acid supplemented in the medium was found to be the dominant fatty acid esterified with retinol. Cultures of stellate cells were then exposed to a physiological concentration (1.3 microM) of radioactive retinol free in solution or bound to retinol-binding protein. With 3-day-old cultures, as well as older cultures, the cellular content of unesterified retinol was 10-20 times higher when free retinol was added compared with addition of retinol bound to retinol-binding protein. However, 2-3-fold as much radioactive retinyl esters were recovered in cells incubated with retinol-retinol-binding protein compared with retinol free in solution. These results show that retinol delivered to stellate cells from retinol-binding protein is preferentially esterified, and that the complex is handled differently to free retinol by the stellate cells.
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Sanchez, Massimo, Francesca Masiello, Valentina Tirelli, Barbara Ghinassi, Carolyn Whitsett, Anna Rita Migliaccio, and Giovanni Migliaccio. "The Final Cellular Output in Human Erythroid Massive Amplification Culture (HEMA) Is Determined by Dynamic Interactions Between Immature and Mature Cell Populations." Blood 114, no. 22 (November 20, 2009): 3156. http://dx.doi.org/10.1182/blood.v114.22.3156.3156.
Abstract:
Abstract Abstract 3156 Poster Board III-93 Hematopoietic progenitor cells cultured with stem cell factor (SCF), interleukin-3 (IL-3) and erythropoietin (EPO) generate erythroid cells that mature in 2 weeks. Addition of dexamethasone and estradiol retards maturation providing the basis for the development of HEMA, a system that generates ex vivo human erythroblasts (EBs) in numbers sufficient for transfusion. In HEMA, EBs at different stages of maturation coexist for > 21 days. FACS analyses for thrombospondin receptor (CD36) and Glycophorin A (CD235a) divides EBs into 4 maturation classes: CD36highCD235aneg (CFU-E) and CD36highCD235alow (pro-EBs) (immature EBs, iEBs); CD36highCD235ahigh (basophilic-polychromatic EBs) (mature EBs) and CD36lowCD235ahigh (orthochromatic EBs) cells. At day 10, 5-19% of the cells are non-erythroid CD36negCD235aneg cells while orthochromatic EBs are barely detectable at any time. The aim of this study was to identify the relative contribution of different populations to the final EBs output of HEMA. The cells present at day 10 in HEMA established with adult mononuclear cells were separated by sorting, labeled with CSFE and cultured for 4 additional days for fate determination (proliferation, maturation and/or death). To identify which population had the ability to generate more EBs, unfractionated cells and cells isolated by sorting (non-erythroid, iEBs and mEBs) were cultured either in colony assay or HEMA. Ninety-three percent of the colony forming ability (all CFU-GM) of unfractionated cells was recovered in the CD36negCD235aneg fraction which did not grow in HEMA. However, iEBs did not contain colony forming cells but generated twice as many EBs as unfractionated cells [3.9±1.0 vs 2.38±0.32 fold increase (FI), respectively). In cultures maintained for 16 days, iEBs sequentially sorted every 2 days generated 5-fold more EBs than the corresponding unfractionated EBs (FI=25 vs 5, respectively). mEBs did not generate colonies or proliferate but died in HEMA. Calculations of hemi CSFE staining decrements indicated that the division index of iEBs and mEBs in the first 24 hrs of HEMA was 1.32±0.08 and 0.4±0.08 (p<0.01), respectively. The higher number of cells generated by iEBs with respect to unfractionated cells suggested that mEBs may inhibit iEBs proliferation. To characterize this effect, iEBs and mEBs were co-cultured for 48 hrs at ratios of 100/0, 5/1, 5/3 and 1/2. One of the two populations was labelled by CSFE staining. To take into account a possible toxic effect of CSFE-labeling, experiments were repeated with either CSFE-labelled iEBs or CSFE-labelled mEBs measuring proliferation/maturation/apoptosis of labeled and unlabelled cells, respectively. iEBs alone increased in numbers by 2-fold over 4 days. By contrast, iEB-derived cells did not increase in number in co-cultures at 5/1 and 5/3 mEB ratios and were undetectable in those at 1/2 mEB ratio. Although cell number did not increase in the co-cultures, the proliferation index (1.3), levels of maturation (50% newly generated mEBs) and apoptosis (barely detectable Annexin Vpos cells) of iEBs in co-cultures remained similar to those observed when these cells were cultured alone. The failure to detect cells after 4 days in iEB/mEB co-cultures at 1/2 ratio suggests that mEBs induce non-apoptotic iEB death. In conclusion, three cells populations were present at day 10 in HEMA of adult mononuclear cells: a CD36negCD235aneg population that contains myeloid progenitor cells and does not generate EBs; iEBs, that generate additional iEBs as well as mEBs; and mEBs that have low proliferation activity, limited life-span and induce non-apototic iEB death. Therefore, the final cellular output of HEMA is determined by a delicate balance between proliferation, maturation and cell death of iEBs and mEBs. Disclosures No relevant conflicts of interest to declare.
46
Gerashchenko, Bogdan I., Igor I. Gerashchenko, Toshikazu Kosaka, and Hiroshi Hosoya. "Stimulatory effect of aerosil on algal growth." Canadian Journal of Microbiology 48, no. 2 (February 1, 2002): 170–75. http://dx.doi.org/10.1139/w01-143.
Abstract:
Unicellular green alga represents not only a convenient model for its biochemical and physiological studies but also a sensitive system to test the effects of various environmental factors. Algae cells of two strains, SA-3 strain (exsymbiotic from Paramecium bursaria) and Chlorella vulgaris c-27, were asynchronously cultured in the presence of 0.01% Aerosil A-300. Aerosil effects on algae were monitored at logarithmic and stationary phases of their growth by flow cytometry and microscopic counting of algal numbers. The growth patterns of algae were evaluated by their forward light scatter versus fluorescence of endogenous chlorophyll (FL3-height) signal distributions. Although aerosil itself did not cause any direct effects on algal morphology, it affected the growth patterns and the numbers of algae of both strains. Their growth patterns were remarkably altered in the late logarithmic phase cultures (6-day cultures). However, a significant increase of cell numbers was found in the stationary phase cultures (9- and 12-day cultures). While C. vulgaris c-27 demonstrated an increase of cell numbers by approximately 11% in the 9- and 12-day cultures, the amounts of SA-3 cells in the 9- and 12-days cultures were increased by 16% and 35%, respectively. Our study shows aerosil in its colloidal form stimulates proliferation of algae mainly via an acceleration of their life cycles. The stimulatory effect of silica on the growth of algae, the mechanism of which remains to be clarified, might have a practical (e.g., ecological) interest for regulation of algal expansion.Key words: aerosil, cloned algae, growth, forward light scatter, chlorophyll fluorescence.
47
Pillai, Manoj M., Mineo Iwata, Debra D. Dahlen, and Beverly Torok-Storb. "Human Marrow Stromal Cells Activate Monocytes to Secrete CXCL7 Peptides, Which Alter the Composition of the Hematopoietic Microenvironment (ME)." Blood 104, no. 11 (November 16, 2004): 668. http://dx.doi.org/10.1182/blood.v104.11.668.668.
Abstract:
Abstract CXCL7 is a 14.0 Kd chemokine produced by megakaryocytes. There are 4 different peptides derived from the pro-peptide by cleavage of the amino terminal, each with distinctive functions: The longest form, LDGF (Leukocyte Derived Growth Factor) is mitogenic for fibroblasts while the smallest, Neutrophil Activating Peptide-2 (NAP-2) inhibits megakaryocytopoiesis. Therefore both forms have, in theory, the potential to influence the composition of the marrow microenvironment. We report here that an additional source of CXCL7 is the monocyte, which upregulates CXCL7 mRNA several fold when stimulated by stromal cells. Briefly, flow sorted CD 14+ monocytes were cultured alone and in co-culture with stromal cell lines HS-5 and HS-27a. Between group comparisons of gene expression profiles were made to identify genes up or down regulated in co-cultures. Expression of CXCL7 was found to be upregulated 25 fold when monocytes were co-cultured with HS-5 and 13 fold with HS-27a as compared to monocytes cultured alone. The array data was confirmed by RT-PCR. Using transwell cultures it was possible to demonstrate that the monocytes and not the stromal cells were responsible for CXCL7 gene expression, which peaked at 6 days. ELISA-based assays detected CXCL7 peptides in the supernatants of monocyte/stroma co-cultures after day 2 of culture. Levels detected ranged from 10 to 50 ng/ml, which are physiologically relevant concentrations. Western Blot analysis showed the 3 day cultures had predominantly LDGF; by day 6 the smaller form, NAP-2 was also present. Since peptide cleavage involves Cathepsin and related proteinases, this delayed appearance of NAP-2 may suggest that the required concentrations of proteinases accumulate with time. The presence of both mRNA and secreted protein in primary long-term cultures of human bone marrow, (peptide levels peaking at 125 ng/ml on day 22) suggest that normal, non-immortalized stroma also induces expression of the CXCL7 gene in the monocytes. Stroma-derived molecules known to stimulate monocytes (IL-1beta, IL-6, GM-CSF and G-CSF) were evaluated for their ability to induce CXCL7 gene expression. Of those tested, only GM-CSF was shown to induce the expression of CXCL7, but at levels far lower than induced by HS-5. As reported previously addition of NAP-2 to CFU-Meg cultures decreased the CFU-Megs by up to 40% despite maximal TPO and the CFU-Megs that did grow were smaller than those in control cultures. Also, addition of flow-sorted monocytes to cultures of primary marrow stroma, not only resulted in CXCL7 gene expression, but significantly increased fibroblast growth in these cultures. In summary our data show that a stroma-derived signal induces the expression of CXCL7 gene products in monocytes. Given that the monocyte is a critical component of the marrow microenvironment we postulate that induction of CXCL7 by stromal signals can affect both structure and function of the marrow microenvironment by increasing fibrosis and decreasing platelet production. Preliminary data from monocytes of 5 patients with Myelodysplastic Syndrome (MDS) suggests that MDS derived monocytes have abnormal patterns of CXCL7 expression. Whether this contributes to the pathophysiology of MDS remains to be determined.
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Orita, Hiroyuki, Manabu Fukasawa, Hideaki Uchino, Kana Fukui, Minoru Kohi, and Masahiko Washio. "Modulation of the viability of immature cardiac myocytes by cardiac fibroblasts after hypothermic preservation—its values as a technique for evaluation of storage solutions." Cardiology in the Young 5, no. 2 (April 1995): 110–17. http://dx.doi.org/10.1017/s1047951100011665.
Abstract:
AbstractWe evaluated the modulation of the viability of immature cardiac myocytes by cardiac fibroblasts after hypothermic preservation using three types of storage solutions—saline, University of Wisconsin solution, and MCDB 107 medium. Cardiac myocytes and fibroblasts were isolated from neonatal rat ventricles, and cultures of myocytes only or co-cultures with fibroblasts (myocyte: fibroblast 2:1) were established. On the fourth day of culture, the cultures were incubated at 4 °C for 6, 12, 18 and 24 hours in the different storage solutions. Enzymes were measured in the storage solutions immediately before and after hypothermic incubation. The cultures were then incubated for an additional 24 hours at 37 °C to evaluate the recovery of the myocyte beating rate. The myocyte beating rate in the co-culture groups showed significantly higher recovery ratios than the corresponding groups in which only myocytes were cultured. Complete recovery was observed in the group co-cultured in MCDB medium 24 hours after hypothermic incubation (83.4% of control—beating rate prior to hypothermic incubation) compared to the other co-cultured groups (15.4, 0%, respectively). Release of enzymes in the co-cultures was significantly suppressed compared to the cultured myocytes, and the greatest suppression was found after 24 hours of incubation in MCDB medium (CPK: 36.6 mIU/flask, LDH: 281.2 mIU/flask) compared to the other two co-cultured groups (CPK: 181.1, 281.1; LDH: 501.7, 773.2). Cardiac fibroblasts diminished myocytic injury after hypothermic preservation using various storage solutions, in which MCDB 107 medium showed the best overall protective effect. Thus, cardiac fibroblasts may play an important role in controlling myocytic viability under various hypothermic conditions.
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Gálfi, P., Susan Neogrády, and G. Gäbel. "Na+/H+ exchange in primary, secondary and n-butyrate-treated cultures of ruminal epithelial cells: Short communication." Acta Veterinaria Hungarica 50, no. 2 (May 1, 2002): 211–15. http://dx.doi.org/10.1556/avet.50.2002.2.10.
Abstract:
Rate of amiloride-sensitive Na+ uptake into cultured rumen epithelial cells was studied in order to clarify the influence of culture conditions on Na+/H+ exchange (NHE). Cell cultures were exposed to Na-n-butyrate or not for seven days or subcultured. On the 14th day of culturing, primary cell cultures without butyrate exposure showed both non-stratified and stratified growth. Na-n-butyrate treated 14-day-old cultures and 3-day-old subcultures contained mostly non-stratified, i.e. non-keratinised cells. Both n-butyrate treatment and subculturing increased total and amiloride-sensitive Na+ uptake. Our results indicate that Na+ uptake via NHE is determined by the amount and the ratio of non-stratified (non-keratinised) cells.
50
Mesel-Lemoine, Mariana, Mustapha Cherai, Sabine Le Gouvello, Maude Guillot, Virginie Leclercq, David Klatzmann, Véronique Thomas-Vaslin, and François M. Lemoine. "Initial depletion of regulatory T cells: the missing solution to preserve the immune functions of T lymphocytes designed for cell therapy." Blood 107, no. 1 (January 1, 2006): 381–88. http://dx.doi.org/10.1182/blood-2005-07-2658.
Abstract:
Abstract We investigated the causes of the altered functionality of T cells cultured under conditions designed for cell and gene therapy and the strategies to prevent their defects. We first showed that human T cells cultured for 6 days with anti-CD3 ± anti-CD28 antibodies and interleukin-2 presented a 50% decrease of their proliferative responses to allogeneic or recall antigens. Similarly, day-6 cultured murine T cells completely lost their capacity to reject allogeneic skin grafts and to provoke graft-versus-host disease (GVHD) when infused into irradiated semi-allogeneic mice. Interestingly, injection of higher amounts of cultured T cells restored GVHD induction. Moreover, depletion of CD25+ cells prior to T-cell cultures can prevent these deficiencies both in mice and humans. Therefore, we demonstrated that culture conditions used for T-cell therapy preferentially activated and expanded regulatory T cells (Treg's). Thus, we showed that dividing cells sorted from T-cell cultures strongly suppressed the proliferation of autologous T cells in response to allogeneic stimulation. An increased detection of Foxp3 at mRNA and protein levels in the cultures confirmed the Treg expansion. Overall, we demonstrate that T-cell cultures promote Treg expansion over effector T cells, leading to deleterious immune functions, and that this imbalance can be prevented by an initial depletion of CD25+ cells.