Contact interactions between epitheliocytes and fibroblasts: Formation of heterotypic cadherin-containing adhesion sites is accompanied by local cytoskeletal reorganization

Contact interactions between different cell types play a number of important roles in development, for example in cell sorting, tissue organization, and ordered migration of cells. The nature of such heterocellular interactions, in contrast to interactions between cells of the same type, remains largely unknown. In this report, we present experimental data examining the dynamics of heterocellular interactions between epitheliocytes and fibroblasts, which express different cadherin cell adhesion molecules and possess different actin cytoskeletal organizations. Our analysis revealed two striking features of heterocellular contact. First, the active free edge of an epitheliocyte reorganizes its actin cytoskeleton after making contact with a fibroblast. Upon contact with the leading edge of a fibroblast, epitheliocytes disassemble their marginal bundle of actin filaments and reassemble actin filaments into a geometric organization more typical of a fibroblast lamella. Second, epitheliocytes and fibroblasts form cell–cell adhesion structures that have an irregular organization and are associated with components of cell adhesion complexes. The structural organization of these adhesions is more closely related to the type of contacts formed between fibroblasts rather than to those between epitheliocytes. Heterotypic epithelio-fibroblastic contacts, like homotypic contacts between fibroblasts, are transient and do not lead to formation of stable contact interactions. We suggest that heterocellular contact interactions in culture may be regarded as models of how tissue systems consisting of epithelia and mesenchyme interact and become organized in vivo.

Apricot culture in California /

Mode of access: Internet.

Interaction of peptides, peptidomimetics and other drug candidates with the DI/tripeptide transport system in intestinal epithelial cells, using the in vitro caco-2 cell culture system

An uptake system was developed using Caco-2 cell monolayers and the dipeptide, glycyl-[3H]L-proline, as a probe compound. Glycyl-[3H]L-proline uptake was via the di-/tripeptide transport system (DTS) and, exhibited concentration-, pH- and temperature-dependency. Dipeptides inhibited uptake of the probe, and the design of the system allowed competitors to be ranked against one another with respect to affinity for the transporter. The structural features required to ensure or increase interaction with the DTS were defined by studying the effect of a series of glycyl-L-proline and angiotensin-converting enzyme (ACE)-inhibitor (SQ-29852) analogues on the uptake of the probe. The SQ-29852 structure was divided into six domains (A-F) and competitors were grouped into series depending on structural variations within specific regions. Domain A was found to prefer a hydrophobic function, such as a phenyl group, and was intolerant to positive charges and H+ -acceptors and donors. SQ-29852 analogues were more tolerant of substitutions in the C domain, compared to glycyl-L-proline analogues, suggesting that interactions along the length of the SQ-29852 molecule may override the effects of substitutions in the C domain. SQ-29852 analogues showed a preference for a positive function, such as an amine group in this region, but dipeptide structures favoured an uncharged substitution. Lipophilic substituents in domain D increased affinity of SQ-29852 analogues with the DTS. A similar effect was observed for ACE-NEP inhibitor analogues. Domain E, corresponding to the carboxyl group was found to be tolerant of esterification for SQ-29852 analogues but not for dipeptides. Structural features which may increase interaction for one series of compounds, may not have the same effect for another series, indicating that the presence of multiple recognition sites on a molecule may override the deleterious effect of anyone change. Modifying current, poorly absorbed peptidomimetic structures to fit the proposed hypothetical model may improve oral bioavailability by increasing affinity for the DTS. The stereochemical preference of the transporter was explored using four series of compounds (SQ-29852, lysylproline, alanylproline and alanylalanine enantiomers). The L, L stereochemistry was the preferred conformation for all four series, agreeing with previous studies. However, D, D enantiomers were shown in some cases to be substrates for the DTS, although exhibiting a lower affinity than their L, L counterparts. All the ACE-inhibitors and β-lactam antibiotics investigated, produced a degree of inhibition of the probe, and thus show some affinity for the DTS. This contrasts with previous reports that found several ACE inhibitors to be absorbed via a passive process, thus suggesting that compounds are capable of binding to the transporter site and inhibiting the probe without being translocated into the cell. This was also shown to be the case for oligodeoxynucleotide conjugated to a lipophilic group (vitamin E), and highlights the possibility that other orally administered drug candidates may exert non-specific effects on the DTS and possibly have a nutritional impact. Molecular modelling of selected ACE-NEP inhibitors revealed that the three carbonyl functions can be oriented in a similar direction, and this conformation was found to exist in a local energy-minimised state, indicating that the carbonyls may possibly be involved in hydrogen-bond formation with the binding site of the DTS.

The evaluation and development of the lead authority partnership scheme as a central intervention strategy for health and safety enforcement by local authorities

The research comprises a suite of studies that examines and develops the Lead Authority Partnership Scheme (LAPS) as a central intervention strategy for health and safety by local authority (LA) enforcers. Partnership working is a regulatory concept that in recent years has become more popular but there has been little research conducted to investigate, explore and evaluate its practical application. The study reviewed two contrasting approaches to partnership working between LAs and businesses, both of which were intended to secure improvements in the consistency of enforcement by the regulators and in the health and safety management systems of the participating businesses. The first was a well-established and highly prescriptive approach that required a substantial resource commitment on the part of the LA responsible for conducting a safety management review (SMR) of the business. As a result of his evaluation of the existing ‘full SMR’ scheme, the author developed a second, more flexible approach to partnership working. The research framework was based upon a primarily qualitative methodology intended to investigate and explore the impact of the new flexible arrangements for partnership working. The findings from this study of the flexible development of the scheme were compared and contrasted with those from studies of the established ‘full SMR’ scheme. A substantial degree of triangulation was applied in an attempt to strengthen validity and broaden applicability of the research findings. Key informant interviews, participant observation, document/archive reviews, questionnaires and surveys all their particular part to play in the overall study. The findings from this research revealed that LAPS failed to deliver consistency of LA enforcement across multiple-outlet businesses and the LA enforced business sectors. Improvement was however apparent in the safety management systems of the businesses participating in LAPS. Trust between LA inspector and safety professional was key to the success of the partnerships as was the commitment of these key individuals. Competition for precious LA resources, the priority afforded to food safety over health and safety, the perceived high resource demands of LAPS, and the structure and culture of LAs were identified as significant barriers to LA participation. Flexible approaches, whilst addressing the resource issues, introduced some fresh concerns relating to credibility and delivery. Over and above the stated aims of the scheme, LAs and businesses had their own reasons for participation, notably the personal development of individuals and kudos for the organisation. The research has explored the wider implications for partnership working with the overall conclusion it is most appropriately seen as a strategic level element within a broader structured intervention strategy.

Development of a co-culture model of the human lungs for toxicity testing and identification of biomarkers of inhalation toxicity

The airway epithelium is the first point of contact in the lung for inhaled material, including infectious pathogens and particulate matter, and protects against toxicity from these substances by trapping and clearance via the mucociliary escalator, presence of a protective barrier with tight junctions and initiation of a local inflammatory response. The inflammatory response involves recruitment of phagocytic cells to neutralise and remove and invading materials and is oftern modelled using rodents. However, development of valid in vitro airway epithelial models is of great importance due to the restrictions on animal studies for cosmetic compound testing implicit in the 7th amendment to the European Union Cosmetics Directive. Further, rodent innate immune responses have fundamental differences to human. Pulmonary endothelial cells and leukocytes are also involved in the innate response initiated during pulmonary inflammation. Co-culture models of the airways, in particular where epithelial cells are cultured at air liquid interface with the presence of tight junctions and differentiated mucociliary cells, offer a solution to this problem. Ideally validated models will allow for detection of early biomarkers of response to exposure and investigation into inflammatory response during exposure. This thesis describes the approaches taken towards developing an in vitro epithelial/endothelial cell model of the human airways and identification biomarkers of response to exposure to xenobiotics. The model comprised normal human primary microvascular endothelial cells and the bronchial epithelial cell line BEAS-2B or normal human bronchial epithelial cells. BEAS-2B were chosen as their characterisation at air liquid interface is limited but they are robust in culture, thereby predicted to provide a more reliable test system. Proteomics analysis was undertaken on challenged cells to investigate biomarkers of exposure. BEAS-2B morphology was characterised at air liquid interface compared with normal human bronchial epithelial cells. The results indicate that BEAS-2B cells at an air liquid interface form tight junctions as shown by expression of the tight junction protein zonula occludens-1. To this author’s knowledge this is the first time this result has been reported. The inflammatory response of BEAS-2B (measured as secretion of the inflammatory mediators interleukin-8 and -6) air liquid interface mono-cultures to Escherichia coli lipopolysaccharide or particulate matter (fine and ultrafine titanium dioxide) was comparable to published data for epithelial cells. Cells were also exposed to polymers of “commercial interest” which were in the nanoparticle range (and referred to particles hereafter). BEAS-2B mono-cultures showed an increased secretion of inflammatory mediators after challenge. Inclusion of microvascular endothelial cells resulted in protection against LPS- and particle- induced epithelial toxicity, measured as cell viability and inflammatory response, indicating the importance of co-cultures for investigations into toxicity. Two-dimensional proteomic analysis of lysates from particle-challenged cells failed to identify biomarkers of toxicity due to assay interference and experimental variability. Separately, decreased plasma concentrations of serine protease inhibitors, and the negative acute phase proteins transthyretin, histidine-rich glycoprotein and alpha2-HS glycoprotein were identified as potential biomarkers of methyl methacrylate/ethyl methacrylate/butylacrylate treatment in rats.

The influence of socio-cultural environments on the performance of nascent entrepreneurs:community culture, motivation, self-efficacy and start-up success

The importance of informal institutions and in particular culture for entrepreneurship is a subject of ongoing interest. Past research has mostly concentrated on cross-national comparisons, cultural values and the direct effects of culture on entrepreneurial behaviour, but in the main found inconsistent results. We add a fresh perspective to this research stream by turning attention to community-level culture and cultural norms. We hypothesize indirect effects of cultural norms on venture emergence: Community-level cultural norms (performance-based culture and socially supportive institutional norms) impact important supply-side variables (entrepreneurial self-efficacy and entrepreneurial motivation) which in turn influence nascent entrepreneurs' success in creating operational ventures (venture emergence). We test our predictions on a unique longitudinal dataset, tracking nascent entrepreneurs' venture creation efforts over a five-year time span, and find evidence supporting them. Our research contributes to a more fine-grained understanding of how culture, in particular perceptions of community cultural norms, influences venture emergence. Based on these findings, we discuss how venture creation efforts can be supported. Our research highlights the embeddedness of entrepreneurial behaviour and its immediate antecedent beliefs in the local, community context. © 2012 Copyright Taylor and Francis Group, LLC.

Reporting and managing risk:a look at current practice at Tesco, RBS, local and central government

These case studies from CIMA highlight the need to embed risk management within more easily understood behaviours, consistent with the overall organisational culture. In each case, some form of internal audit team provides either an oversight function or acts as an expert link in that feedback loop. Frontline staff, managers and specialists should be completely aligned on risk, in part just to ensure that there is a consistency of approach. They should understand instinctively that good performance includes good risk management. Tesco has continued to thrive during the recession and remains a robust and efficient group of businesses despite the emergence of potential threats around consumer spending and the supply chain. RBS, by contrast, has suffered catastrophic and very public failures of risk management despite a large in-house function and stiff regulation of risk controls. Birmingham City Council, like all local authorities, is adapting to more commercial modes of operation and is facing diverse threats and opportunities emerging as a result of social change. And DCMS, like many other public sector organisations, has to handle an incredibly complex network of delivery partners within the context of a relatively recent overhaul of central government risk management processes. Key Findings: •Risk management is no longer solely a financial discipline, nor is it simply a concern for the internal control function. •Where organisations retain a discrete risk management cadre – often specialists at monitoring and evaluating a range of risks – their success is dependent on embedding risk awareness in the wider culture of the enterprise. •Risk management is most successful when it is explicitly linked to operational performance. •Clear leadership, specific goals, excellent influencing skills and open-mindedness to potential threats and opportunities are essential for effective risk management. •Bureaucratic processes and systems can hamper good risk management – either as a result of a ‘box-ticking mentality’ or because managers and staff believe they do not need to consider risk themselves.

Culture, ethnicity and migration after communism:the Pontic Greeks

This book addresses the issue of emerging transnationalism in the conditions of post-socialism through focussing on migrants’ identity as a social construction resulting from their experience of the ‘transnational circuit of culture’ as well as from post-Soviet shifts in political and economic conditions in their home regions. Popov draws upon ethnographic research conducted among Greek transnational migrants living on the Black Sea coast and in the North Caucasus regions of Russia who have become involved in extensive cross-border migration between the former Soviet Union (the Russian Federation, Kazakhstan and Georgia) and Greece (as well as Cyprus). It is estimated that more than 150,000 former Soviet citizens of Greek origin have resettled in Greece since the late 1980s. Yet, many of those who emigrate do not cut their connections with the home communities in Russia but instead establish their own transnational circuit of travel between Greece and Russia. This study demonstrates how migrants employ their ethnicity as symbolic capital available for investment in profitable transnational migration. Simultaneously they rework their practices of family networking, property relations and political participation in a way which strengthens their attachment to the local territory. The findings presented in the book imply that the social identities, economic strategies, political practices and cultural representation of the Russian Greeks are all deeply embedded in the shifting social and cultural landscape of post-Soviet Russia and extensively influenced by the global movement of ideas, goods and people.

Determinants of local government pension funding: Empirical evidence from Florida

The financial community is well aware that continued underfunding of state and local government pension plans poses many public policy and fiduciary management concerns. However, a well-defined theoretical rationale has not been developed to explain why and how public sector pension plans underfund. This study uses three methods: a survey of national pension experts, an incomplete covariance panel method, and field interviews.^ A survey of national public sector pension experts was conducted to provide a conceptual framework by which underfunding could be evaluated. Experts suggest that plan design, fiscal stress, and political culture factors impact underfunding. However, experts do not agree with previous research findings that unions actively pursue underfunding to secure current wage increases.^ Within the conceptual framework and determinants identified by experts, several empirical regularities are documented for the first time. Analysis of 173 local government pension plans, observed from 1987 to 1992, was conducted. Findings indicate that underfunding occurs in plans that have lower retirement ages, increased costs due to benefit enhancements, when the sponsor faces current year operating deficits, or when a local government relies heavily on inelastic revenue sources. Results also suggest that elected officials artificially inflate interest rate assumptions to reduce current pension costs, consequently shifting these costs to future generations. In concurrence with some experts there is no data to support the assumption that highly unionized employees secure more funding than less unionized employees.^ Empirical results provide satisfactory but not overwhelming statistical power, and only minor predictive capacity. To further explore why underfunding occurs, field interviews were carried out with 62 local government officials. Practitioners indicated that perceived fiscal stress, the willingness of policymakers to advance funding, bargaining strategies used by union officials, apathy by employees and retirees, pension board composition, and the level of influence by internal pension experts has an impact on funding outcomes.^ A pension funding process model was posited by triangulating the expert survey, empirical findings, and field survey results. The funding process model should help shape and refine our theoretical knowledge of state and local government pension underfunding in the future. ^

Without women: Masculinities, gay male sexual culture and sexual behaviors in Miami, Florida

Recent studies have reported alarmingly high rates of HIV infection and risky sexual behaviors among gay men in Miami, Florida. Previous research has suggested that the risky sexual behaviors of many gay men reflect the pursuit of intimacy and love, and that barriers to intimate relationships among gay men may stem from traditional masculinity norms. This dissertation examines the meanings which gay men ascribe to their sexual behaviors, as well as the intersections of those meanings with both traditional masculinity constructions and Miami's gay male sexual culture. ^ The study is based upon participant observation, print media content analysis, surveys and ethnographic interviews of a purposive snowball sample of 30 Cuban American, Puerto Rican, African American and Anglo gay men who reside in Miami-Dade County, Florida. Analysis of research questions was accomplished through grounded theory methods and descriptive and non-parametric statistics, including Pearson chi-square, Fisher's Exact and Mann-Whitney U tests. ^ The study shows that culturally-specified masculinity norms vary in the relative importance ascribed to heterosexual prowess, economic providership and competitiveness. These cultural differences appear important not only to the timing of sexual awareness and to the strength of homosexual stereotyping as effeminacy, but also to men's strategies in coming out as gay. The meanings men attributed to their sexual behaviors were, however, constructed in response to both inherited masculinity norms and the hypermasculine structure of Miami's gay male sexual culture. In addition to providing an ethnographic account of this subculture, the study elaborates men's issues relative to casual sex and committed relationships. Unprotected anal intercourse with casual partners during the previous twelve months was associated with growing up without one's father in the home, having been teased for effeminacy during childhood, being defensive about one's masculinity, not trusting men, having been cheated on by boyfriends, and believing that long-term gay male relationships are problematic. ^ It is concluded that the continuing epidemic of HIV infections among local gay men, as well as the hypermasculine form of the gay sexual subculture itself, are nihilistic symptoms embedded in the masculinist gender structure of the larger society. ^

The transition movement and food sovereignty: from local resilience to global engagement in food system transformation

The emergence of grassroots social movements variously preoccupied with a range of external threats, such as diminishing supplies of fossil energy or climate change, has led to increased interest in the production of local food. Drawing upon the notion of cognitive praxis, this article utilises transition as a trajectory guided by an overarching cosmology that brings together a broad social movement seeking a more resilient future. This ‘grand narrative’ is reinforced by ‘transition movement intellectuals’ who serve to shape an agenda of local preparedness in the face of uncertainty, rather than structural analysis of the global system. In this context, growing and producing food offers important multi-functional synergies by reconnecting people to place and its ecological endowments and serves to provide a vital element in civic mobilisation. Yet, local food could also become a means to build international solidarity in defence of food sovereignty and establish a global coalition opposed to the corporate agri-food agenda of biotechnologies, land grabbing and nutritional impoverishment.

Beyond the Convent Walls: The Local and Japan-wide Activities of Daihongan’s Nuns in the Early Modern Period (c. 1550–1868)

This dissertation examines the social and financial activities of Buddhist nuns to demonstrate how and why they deployed Buddhist doctrines, rituals, legends, and material culture to interact with society outside the convent. By examining the activities of the nuns of the Daihongan convent (one of the two administrative heads of the popular pilgrimage temple, Zenkōji) in Japan’s early modern period (roughly 1550 to 1868) as documented in the convent’s rich archival sources, I shed further light on the oft-overlooked political and financial activities of nuns, illustrate how Buddhist institutions interacted with the laity, provide further nuance to the discussion of how Buddhist women navigated patriarchal sectarian and secular hierarchies, and, within the field of Japanese history, give voice to women who were active outside of the household unit around which early modern Japanese society was organized.

Zenkōji temple, surrounded by the mountains of Nagano, has been one of Japan’s most popular pilgrimage sites since the medieval period. The abbesses of Daihongan, one Zenkōji’s main sub-temples, traveled widely to maintain connections with elite and common laypeople, participated in frequent country-wide displays of Zenkōji’s icon, and oversaw the creation of branch temples in Edo (now Tokyo), Osaka, Echigo (now Niigata), and Shinano (now Nagano). The abbesses of Daihongan were one of only a few women to hold the imperially sanctioned title of eminent person (shōnin 上人) and to wear purple robes. While this means that this Pure Land convent was in some ways not representative of all convents in early modern Japan, Daihongan’s position is particularly instructive because the existence of nuns and monks in a single temple complex allows us to see in detail how monastics of both genders interacted in close quarters.

This work draws heavily from the convent’s archival materials, which I used as a guide in framing my dissertation chapters. In the Introduction I discuss previous works on women in Buddhism. In Chapter 1, I briefly discuss the convent’s history and its place within the Zenkōji temple complex. In Chapter 2, I examine the convent’s regular economic bases and its expenditures. In Chapter 3, I highlight Daihongan’s branch temples and discuss the ways that they acted as nodes in a network connecting people in various areas to Daihongan and Zenkōji, thus demonstrating how a rural religious center extended its sphere of influence in urban settings. In Chapter 4, I discuss the nuns’ travels throughout the country to generate new and maintain old connections with the imperial court in Kyoto, confraternities in Osaka, influential women in the shogun’s castle, and commoners in Edo. In Chapter 5, I examine the convent’s reliance upon irregular means of income such as patronage, temple lotteries, loans, and displays of treasures, and how these were needed to balance irregular expenditures such as travel and the maintenance or reconstruction of temple buildings. Throughout the dissertation I describe Daihongan’s inner social structure comprised of abbesses, nuns, and administrators, and its local emplacement within Zenkōji and Zenkōji’s temple lands.

Exploring these themes sheds light on the lives of Japanese Buddhist nuns in this period. While the tensions between freedom and agency on the one hand and obligations to patrons, subordination to monks, or gender- and status-based restrictions on the other are important, and I discuss them in my work, my primary focus is on the nuns’ activities and lives. Doing so demonstrates that nuns were central figures in ever-changing economic and social networks as they made and maintained connections with the outside world through Buddhist practices and through precedents set centuries before. This research contributes to our understanding of nuns in Japan’s early modern period and will participate in and shape debates on the roles of women in patriarchal religious hierarchies.

Local Seafood Availability in San Diego, California Seafood Markets

Like many developed coastal cities, San Diego, California has strong geographic and recreational ties to the adjacent ocean, but weak culinary ones. Less than 10% of the seafood consumed in the U.S., and San Diego in particular, is domestic. The popularity and abundance of farmers’ markets and other local markets in San Diego indicates an interest among producers and the public alike in cultivating local, diverse food systems, but this trend has been slower to catch on for seafood. The goal of this project was, therefore, to define and begin to understand the influences on the patterns of locally sourced, domestic seafood availability in San Diego. This study focused on seafood availability in seafood markets including researching market websites and contacting seafood counter managers to determine the general frequency (consistent, occasional, none) at which the markets sold seafood produced by San Diego fishermen or aquafarmers. Seafood market locations were mapped, and demographic and spatial information was gathered for each market’s zip code. The results of the study revealed that only 8% of San Diego’s 86 seafood markets consistently carried San Diego-sourced seafood, and 14% of markets carried it on occasion. Increased density of these local seafood markets was correlated with proximity to the coast, with almost 80% of the markets located within 2 km of the coast. Neither per capita income nor racial diversity was correlated with local seafood market density, indicating that factors contributing to coastal isolation matter more than wealth or diversity in determining where local seafood is sold. The geographic disparity in local seafood availability may be due to a variety of factors, including a small fishing fleet, prevalence of imported seafood, limited waterfront and urban infrastructure needed to support a local seafood system, and a lack of public awareness about local fisheries. Information gleaned from this study can inform further investigation into the influences on local, equitable seafood systems, as well as help consumers, producers and marketers to make informed decisions about seafood purchases and marketing efforts.