bluuebunny/crossref_metadata_2025 huggingface dataset:
Primitive synteny of vertebrate major histocompatibility complex class I and class II genes
Authors: Ohta Yuko , Okamura Kazuhiko , McKinney E. Churchill , Bartl Simona , Hashimoto Keiichiro , Flajnik Martin F.
Published: July 2002

Major histocompatibility complex (MHC) class I and class II molecules bind to and display peptidic antigens acquired from pathogens that are recognized by lymphocytes coordinating and executing adaptive immune responses. The two classes of MHC proteins have nearly identical tertiary structures and were derived from a common ancestor that probably existed not long before the emergence of the cartilaginous fish. Class I and class II genes are genetically linked in tetrapods but are not syntenic in teleost fish, a phylogenetic taxon derived from the oldest vertebrate ancestor examined to date. Cartilaginous fish (sharks, skates, and rays) are in the oldest taxon of extant jawed vertebrates; we have carried out segregation analyses in two families of nurse sharks and one family of the banded houndshark that revealed a close linkage of class IIα and β genes both with each other and with the classical class I (class Ia) gene. These results strongly suggest that the primordial duplication giving rise to classical class I and class II occurred in cis, and the close linkage between these two classes of genes has been maintained for at least 460 million years in representatives of most vertebrate taxa.
Urothelial function reconsidered: A role in urinary protein secretion
Authors: Deng Fang-Ming , Ding Mingxiao , Lavker Robert M. , Sun Tung-Tien
Published: July 2002

Mammalian bladder epithelium functions as an effective permeability
barrier. We demonstrate here that this epithelium can also function as
a secretory tissue directly involved in modifying urinary protein
composition. Our data indicate that normal bovine urothelium
synthesizes, as its major differentiation products, two well-known
proteases: tissue-type plasminogen activator and urokinase, as well as
a serine protease inhibitor, PP5. Moreover, we demonstrate that the
urothelium secretes these proteins in a polarized fashion into the
urine via a cAMP- and calcium-regulated pathway. Urinary plasminogen
activators of ruminants are therefore urothelium derived rather then
kidney derived as in some other species; this heterogeneity may have
evolved in response to different physiological or dietary factors. In
conjunction with our recent finding that transgenic mouse urothelium
can secrete ectopically expressed human growth hormone into the urine,
our data establish that normal mammalian urothelium can function not
only as a permeability barrier but also as a secretor of urinary
proteins that can play physiological or pathological roles in the
urinary tract.
CpG island protects Rous sarcoma virus-derived vectors integrated into nonpermissive cells from DNA methylation and transcriptional suppression
Authors: Hejnar Jir̆í , Hájková Petra , Plachý Jir̆í , Elleder Daniel , Stepanets Volodymyr , Svoboda Jan
Published: July 2002

CpG islands are important in the protection of adjacent
housekeeping genes from
de novo
DNA methylation and for
keeping them in a transcriptionally active state. However, little is
known about their capacity to protect heterologous genes and assure
position-independent transcription of adjacent transgenes or retroviral
vectors. To tackle this question, we have used the mouse
aprt
CpG island to flank a Rous sarcoma virus
(RSV)-derived reporter vector and followed the transcriptional activity
of integrated vectors. RSV is an avian retrovirus which does not
replicate in mammalian cells because of several blocks at all levels of
the replication cycle. Here we show that our RSV-derived reporter
proviruses linked to the mouse
aprt
gene CpG island
remain undermethylated and keep their transcriptional activity after
stable transfection into both avian and nonpermissive mammalian cells.
This effect is most likely caused by the protection from
de
novo
methylation provided by the CpG island and not by
enhancement of the promoter strength. Our results are consistent with
previous finding of CpG islands in proximity to active but not inactive
proviruses and support further investigation of the protection of the
gene transfer vectors from DNA methylation.
The HIV-1 regulatory proteins Tat and Rev are frequently targeted by cytotoxic T lymphocytes derived from HIV-1-infected individuals
Authors: Addo Marylyn M. , Altfeld Marcus , Rosenberg Eric S. , Eldridge Robert L. , Philips Mary N. , Habeeb Kathleen , Khatri Ashok , Brander Christian , Robbins Gregory K. , Mazzara Gail P. , Goulder Philip J. R. , Walker Bruce D. ,   the HIV Controller Study Collaboration¶
Published: July 2002

The HIV-1 regulatory proteins Rev and Tat are expressed early in the virus life cycle and thus may be important targets for the immune control of HIV-1-infection and for effective vaccines. However, the extent to which these proteins are targeted in natural HIV-1 infection as well as precise epitopes targeted by human cytotoxic T lymphocytes (CTL) remain to be defined. In the present study, 57 HIV-1-infected individuals were screened for responses against Tat and Rev by using overlapping peptides spanning the entire Tat and Rev proteins. CD8+ T cell responses against Tat and Rev were found in up to 19 and 37% of HIV-1-infected individuals, respectively, indicating that these regulatory proteins are important targets for HIV-1-specific CTL. Despite the small size of these proteins, multiple CTL epitopes were identified in each. These data indicate that Tat and Rev are frequently targeted by CTL in natural HIV-1 infection and may be important targets for HIV vaccines.
Public Space, Urban Space and Electronic Space: Would the Real City Please Stand Up?
Authors: Crang Mike
Published: July 2002

Commentary around the electronic media has raised issues of political action, community formation and changing identities. This paper explores how the notions of 'public space' can inform this debate over electronic media. It examines the metaphorical adoption of urban models to look at electronic sociality and suggests four principle approaches: cities set in or against world flows, suburbanised telecities, communitarian visions and accounts that appeal to a renewed public sphere. The paper examines how these share many assumptions. However, instead of trying to sift these metaphors by contrasting them to a purported real world, the paper examines how they shape an electronic architecture. Spatial metaphors and electronic practices are seen as entangled and shaping each other. The paper suggests that the different metaphors for the city reflect a range of anxieties about and desires for urban life. In this sense, the 'real' city is the indefinable complexity and folding of spaces—lying outside the visualisations offered of cyberspace.
The Social Relations of Organisational Activity and the New Local Governance in the UK
Authors: Raco Mike
Published: July 2002

The nature and form of local governance in the UK became the focus for a wide range of academic and policy debates during the 1980s and 1990s. The growth of non-elected localist agencies working alongside local authorities had a major influence on the effectiveness of, and the local policy-making processes behind, policy areas such as urban regeneration. This paper contributes to these debates by arguing that one often neglected or underemphasised aspect of our understanding of local governance is that of the social relations of organisational activity. Drawing on organisational theories and linking them to contemporary debates on local governance, the paper suggests that an exploration of the internal dynamics of non-elected institutions, working in and though wider social, economic and political contexts, provides a more sophisticated understanding of contemporary local political processes than studies which focus on the powers, resources and responsibilities of organisations per se. The study suggests that, in the context of partnership-building and institutional co-operation at the local level, the types of organisational dynamics highlighted in the paper may take on even greater significance in future.
Inner-city and Suburban Labour Markets in a Major English Conurbation: Processes and Policy Implications
Authors: Lawless Paul
Published: July 2002

Unemployment is one of the key characteristics of Britain's inner cities. This paper examines labour market processes and their policy implications within an inner-city and a suburban area of an economically depressed city in England, Sheffield. Five major themes are examined: markedly contrasting unemployment rates within the two areas and their policy implications; job barriers, aspirations and opportunities; skills attainment and training issues; differential unemployment rates affecting specific socio-economic groups in each locality; and questions surrounding poverty.
Commuting and Gender in a Lifestyle Perspective
Authors: Camstra Ronald
Published: July 2002

This article examines gender differences in the context of the relationship between residential relocation and commuting distance. Traditional theories stress women's higher sensitivity to distance, and expect women to quit their jobs more often in the event of a residential move. This appears to be true. However, the gender difference in quitting is constant by distance. Also, women who move over very short distances quit their jobs more often than men. Furthermore, women who remained working at the same job saw their commuting distance increase even more substantially than that of men. Therefore, there is no evidence for the sensitivity hypothesis. Instead, significant effects of lifestyle are found. In some ways, gender differences are almost absent for the most 'modern' lifestyle group.
Institutional Responses to the UK Housing Market Recession
Authors: Stephens Mark
Published: July 2002

Since 1990 the UK housing market has been in recession, with protracted periods of nominal house price falls. While demand-side factors are important in explaining this phenomenon, this article examines the responses of the major institutions to the recession, covering the period up to the autumn of 1995. The institutions examined are lenders, insurance companies and the government (including regulators). The analysis identifies 15 impacts resulting from their behaviour. Of these, eight were pro-cyclical, five were counter-cyclical and two were broadly neutral. The key role of the government in arranging counter-cyclical behaviour is identified, although this should be balanced against other policy objectives, such as the removal of distortions from the housing market.
Changing Spatial Distribution and Determinants of Land Development in Chinese Cities in the Transition from a Centrally Planned Economy to a Socialist Market Economy: A Case Study of Guangzhou
Authors: Wu Fulong , Yeh Anthony Gar-On
Published: July 2002

The spatial distribution and locational characteristics of land development have changed dramatically in Chinese cities since the land reform of 1987 which allowed the paid transfer of land-use rights—i.e. land leasing. This has led to the rapid transformation of the urban spatial structure of Chinese cities. There is an urgent need to study the general trend of such changes and their policy implications. However, due to the lack of data, such investigations lag far behind the rapid land development in Chinese cities. This paper attempts to examine the new spatial pattern of land development in Chinese cities and its determinants by studying land development in Guangzhou before (1979-87) and after (1987-92) the land reform by analysing data obtained from aerial photographs with the aid of GIS techniques. The determinants of land development are analysed using a logistic regression model. It is found that there has been significant acceleration of urban redevelopment and urban sprawl in Guangzhou since the adoption of the new land-leasing system in 1987. The changing spatial distributions and determinants of land development suggest the emergence of new locational behaviours of land development in Chinese cities in the transition from a centrally planned economy to a socialist market economy.
Sydney, Australia: A Global City? Testing the Social Polarisation Thesis
Authors: Baum Scott
Published: July 2002

It has been argued that the global city-social polarisation thesis put forward by scholars looking at North American cities cannot be easily transferred to global cities in other parts of the world. Recent research has illustrated that whilst there may be some change in levels of social polarisation in global cities outside the US, the form, structure and causes of social polarisation are different. This paper extends the debate by looking at Sydney, Australia. It is argued that whilst changes in occupational structure and income polarisation are partially explained by economic restructuring associated with globalisation, global processes alone cannot fully explain these changes. The paper points to the significance of the inclusion of factors such as the unemployed, the gendered structure of occupations and migration.
Housing Finance and Urban Infrastructure Finance
Authors: Kim Kyung-Hwan
Published: July 2002

Financing issues received major attention in the discussions of human settlement development policy through the preparatory process for Habitat II and at the Conference itself. This paper presents an overview of the existing knowledge and policy experience in housing finance and urban infrastructure finance taking note of changes in economic environment such as globalisation, financial liberalisation and decentralisation as well as progress made over the past two decades using cases of good practice. Despite the positive developments such as micro-finance and private-public partnerships, the financing challenge of today is no smaller than before. It includes how to tap the enormous wealth generated through continued urbanisation to improve the conditions in housing and basic services of all income groups and how to translate the willingness to pay of citizens into efficient, accountable and responsive service delivery. The solution requires not only financial innovation but also good governance and political commitment.
Housing Price, Land Supply and Revenue from Land Sales
Authors: Tse Raymond Y. C.
Published: July 2002

In a recent study, Peng and Wheaton empirically examine the effect of restrictive land supply on Hong Kong house prices. However, we argue that the impact of the supply of new land by the Hong Kong government is not as important as they maintain in accounting for the volatile house prices in Hong Kong. We show that there is no causal relationship between land supply and housing prices. Our estimations, based on the annual data of Hong Kong's public land sales, find that the government acts to maximise land revenue. Maximisation of revenue from land sales is, however, consistent with the efficient allocation of resources. We also show that the amount of land sales by the government and land in developers' land banks tend to decrease when market interest rates increase. Land banking behaviour is governed by economic conditions. Long-term land holding costs should cover interest costs.
Socio-spatial Dynamics: Ethnic Divisions of Mobility and Housing in post-Palme Sweden
Authors: Andersson Roger
Published: July 2002

The aim of this article is to analyse ethnic residential segregation in Sweden, both theoretically and empirically. It is argued that ethnic residential segregation, although it is an intra-urban phenomenon, must be viewed in a wider geographical perspective. The three major competing frameworks for analysing ethnic segregation are discussed: the cultural, the cultural racist and the structural approaches. Data suggest that all these three approaches may have relevance, but emphasis has to be put on the two latter explanatory frameworks. A dynamic approach is proposed which should focus on the key role of ethnic/race selective migration in shaping the ethnically partitioned city. This approach emphasises the importance of analysing the effect side of the segregated city and it includes an analysis of the institutional framework. Finally, and based on the proposed model, the role of politics in combating the contemporary social situation in so-called immigrant-dense neighbourhoods is discussed.
Developmental trends in children’s feeling-of-knowing judgements
Authors: Lockl Kathrin , Schneider Wolfgang
Published: July 2002

The main goal of this study was to examine developmental trends in the accuracy of feeling-ofknowing (FOK) judgements given that previous work in this issue had yielded inconsistent results. Although most research conducted with preschoolers and young school children has reported increasing accuracy with increasing age, a methodologically improved study (Butterfield, Nelson, & Peck, 1988) did not confirm such a trend. The present study was based on such an improved design but included different age groups (7-, 8-, 9-, and 10-year-olds). A second goal was to explore the basis of FOK judgements, by comparing the traditional “trace-based” view with the “trace accessibility” model developed by Koriat (1993). Whereas the former assumes a two-stage process of monitoring and retrieval, the latter proposes that FOK judgements are based on retrieval attempts and determined by the amount of information that can be spontaneously generated, regardless of its correctness. As a main result, no developmental trends in the accuracy of FOK judgements were found. FOK accuracy was generally low but above chance for all age groups. Furthermore, the findings support the “trace accessibility” model in that dissociations between feeling of knowing and knowing could be demonstrated. That is, although FOK judgements regarding subsequent item recognition were generally high whenever an answer could be generated and much lower when no answer was generated at free retrieval, regardless of its correctness, recognition performance was comparatively low for previous nonretrieved and incorrectly remembered items and close to ceiling for correctly remembered items.
Stages of Progression in Noncoital Sexual Interactions among Young Adolescents: An Application of the Mokken Scale Analysis
Authors: Jakobsen Reidar
Published: July 2002

This study attempted to test empirically the tenability of the assumption of a sequential pattern of involvement in noncoital sexual interactions among white Western adolescents, by following a cohort between the ages of 13 and 16 years. The results documented that even among 13-year-old adolescents, one can already examine variations in noncoital sexual interaction, and that the various items of noncoital sexual interactions can be reliably measured. The Mokken scale analysis yielded empirical evidence of a sequential pattern of involvement in such behaviour among adolescents. Furthermore, it suggested that in early adolescence, the development of intimate sexual relationships is a slow, experimental, and cumulative process. For most adolescents, this process involves experimentation and the passage of time. The five items of noncoital sexual interactions form a strong Mokken scale, and were significant predictors of the onset of sexual intercourse.
Japanese Children’s Action-Control Beliefs about School Performance
Authors: Karasawa Mayumi , Little Todd D. , Miyashita Takahiro , Mashima Mari , Azuma Hiroshi
Published: July 2002

Guided by research on German, Russian, and American children, we tested whether the tripartite action-theory model of children’s psychological control generalises to Japanese children (grades 2-6, N 817). Specifically, we used the Control, Agency, and Means-ends Interview (CAMI) to assess whether Japanese children’s self-related agency beliefs, general control expectancies, and causality-relate d means-ends beliefs about their school performance are similar to those of children from other sociocultural contexts. The CAMI has shown strong cross-cultural validity, but it has not been tested in Japanese children. Because the CAMI measurement structure generally validated in this sample and the resulting action-control constructs showed many inter-cultural similarities, we concluded that the action-control beliefs generalise to Japanese children. The similarities likely reflect inter-cultural commonalities in teaching formats and everyday conceptions of performance in formal schooling contexts. In addition to these important similarities, however, we found inter-cultural differences in the self-related agency beliefs (i.e. patterns that were specific to this Japanese sample). For example, the role of luck and the relations between effort and ability showed unique patterns in these children (e.g. lower correlations than in other sociocultural settings).
Shaping a Regional Vision: the Case of Northern Ireland
Authors: McEldowney Malachy , Sterrett Ken
Published: July 2002

The theme of this paper is the preparation of a new region al plan for Northern Ireland, known as Shaping Our Future: a Region al Strategic Framework (RSF) and the public consultation exercise which underpinned its development. The paper attempts to provide a reflective con textualized account of the development of a ‘new’ model of participation an d its operation in practice. Arguably, the particular political circumstances in Northern Ireland provide an interesting case study of issues and tensions that connect to wider debates about the shaping of strategic planning policy and the growing demands for the development of participatory democracy.
The Role of Small Towns in the Local Economy and Some Implications for Development Policy
Authors: Courtney Paul , Errington Andrew
Published: July 2002

Over recent years, socio-economic restructuring has tended to undermine the tradition al functions of small rural towns. Empirical evidence also suggests that significant proportion s of economic transactions are carried out through the region al, national an d international economy. Establishing the degree to which a settlement is integrated into its locality provides an indication of its contemporary function s an d potential role in rural development. In this paper, reliable measures of economic linkages are developed, and it is shown that the strength of local integration is a function of the existing economic structure of the town as well as its proximity to metropolitan centres.
Inhibitory effects of Spirulina in zymosan‐induced arthritis in mice
Authors: Remirez Diadelis , González Ricardo , Merino Nelson , Rodriguez Sandra , Ancheta Odelsa
Published: July 2002

The anti‐inflammatory effect of microalgae Spirulina was studied in zymosan‐induced arthritis in mice. Four days after the intra‐articular injection of zymosan (15 mg/ml), Spirulina (100 and 400 mg/kg per‐orally) was administered to animals for 8 days. The mice were than killed and β‐glucuronidase was measured in the synovial fluid. Each knee joint was totally removed for histopathological studies. Spirulina significantly reduced the levels of β‐glucuronidase that had been increased by zymosan. Histopathological and ultrastructural studies showed inhibition of the inflammatory reaction, whereas no destruction of cartilage, well‐preserved chondrocytes, and normal rough endoplasmic reticulum and mitochondria were seen. The anti‐arthritic effect exerted by Spirulina as shown in this model may be at least partly due to the previously reported anti‐inflammatory and antioxidative properties of its constituent, phycocyanin. To our knowledge, this is the first report on the anti‐inflammatory effect of Spirulina in an experimental model of arthritis.
Permeability of blood‐tear barrier to fluorescein and albumin after application of platelet‐activating factor to the eye of the guinea pig
Authors: Van Delft J. L. , Meijer F. , Van Best J. A. , Van Haeringen N. J.
Published: July 2002

One of the inflammatory responses of the eye to local application of platelet‐activating factor (PAF) is oedema of the conjunctiva, caused by extravasation of plasma. Aim of the study was to investigate if fluorescein would leak from the blood into the tears together with plasma protein after application of PAF to the eye. Fluorescein was given intraperitoneally 30 min prior to application of 25 μl of 0.1% solution of PAF. Thirty min after PAF the tear film was collected by washing the surface of the eye with 25 μl of phosphate buffered saline (PBS). Fluorescein in eye washings and in plasma was measured by fluorophotometry and albumin by immunodiffusion. Both fluorescein and albumin appeared in a related fashion in tears, being absent in washings of placebo‐treated control eyes. Extravasation of fluorescein can be used as a measure for plasma leakage in the conjunctiva with the advantage over the Evans Blue method that the former is a non‐invasive method.
Leukocyte activation by (1→3)‐β‐D glucans
Authors: Adachi Yoshiyuki , Okazaki Mitsuhiro , Ohno Naohito , Yadomae Toshiro
Published: July 2002

We studied the activities of several kinds of β‐glucans, including sonifilan, grifolan, Sclerotinia sclerotiorum glucan, laminarin and zymosan, on macrophages. Preculture of macrophages with inactive β‐glucans rendered the cells unresponsive to subsequent stimulation with grifolan, suggesting a specific pathway in the β‐glucan structure. The importance of protein C and phosphorylation of mitogen‐activated protein kinase was demonstrated in the activation with grifolan or zymosan. Immunoprecipitation of complement receptor (CR3), coprecipitated other proteins carrying phosphotyrosine residues in stimulation with grifolan. These data suggest that protein kinase C and tyrosine kinases are essential for signal transduction, and that CR3 might participate in the activation through interaction with other intracellular proteins.
An immunoassay for the measurement of (1→3)‐β‐D‐glucans in the indoor environment
Authors: Douwes Jeroen , Doekes Gert , Montijn Roy , Heederik Dick , Brunekreef Bert
Published: July 2002

An inhibition enzyme immunoassay was developed for quantitation of (1→3)‐β‐D‐glucans in the indoor environment. Immunospecific rabbit antibodies were produced by immunization with bovine serum albuminconjugated laminarin.The laminarin calibration curve ranged from 40 to 3000 ng/ml.Another (1→3)‐β‐D‐glucan (curdlan) showed a similar inhibition curve, but was less reactive on a weight basis. Pustulan, presumed to be (1→3)‐β‐D‐glucan, also showed immunoreactivity in the assay. Control experiments indicated that this was due to (1→3)‐β‐D‐glucan structures. Other non‐(1→3)‐β‐D‐glucan polysaccharides did not react. (1→3)‐β‐Dglucan was detectable in dust from a variety of occupational and environmental settings. We conclude that the new assay offers a useful method for indoor (1→3)‐β‐Dglucan exposure assessment.
Curdlan sulphate modulates protein synthesis and enhances NF‐κB and C/EBP binding activity in HepG2 cells
Authors: Guzdek A. , Rokita H.
Published: July 2002

In human hepatoma HepG2 cell line curdlan sulphate enhances basal and interleukin‐6‐stimulated fibrinogen and antichymotrypsin (ACT) synthesis, slightly increases basal ceruloplasmin production and exerts only minor effects on alpha‐1‐proteinase inhibitor and transferrin. Curdlan sulphate may, at least in part, affect protein synthesis at a pretranslational level, as the expression of ACT mRNA was found to be increased, whereas intracellular enzyme, manganese superoxide dismutase mRNA level was decreased in the cell culture treated with curdlan sulphate. Gel mobility shift analysis revealed that curdlan sulphate increases the DNA binding activity of NF‐κB and C/EBP, suggesting that these transcription factors may participate in the regulatory effects of curdlan sulphate in HepG2 cells.
Effects of anabolic steroids on acute phase responses in intra‐abdominal sepsis
Authors: Mealy K. , Clooney A. , Marks P. , Hennessy T. , Jackson J.
Published: July 2002

The acute phase response is an important adaptive response to sepsis and injury. As anabolic steroids increase protein synthesis we postulated that these agents might also increase hepatic acute phase protein synthesis. Male Wistar rats were pretreated with testosterone or danazol for 48 h prior to caecal ligation and puncture (CLP). Thirty‐six h following surgery the animals were killed and blood taken for full blood count, total protein, albumin, α, β and γ globulin fractions on serum electrophoresis, complement C3 and transferrin levels. Danazol increased the α1, α2 and β1 globulin serum protein fractions in comparison with no surgery and CLP alone groups. These results indicate that danazol increases plasma acute phase proteins, as measured by electrophoresis, in this model of intra‐abdominal sepsis.
Regulation of Inflammatory Responses to Bordetella Pertussis by NG‐Monomethyl‐L‐Arginine in Mice Intranasally Infected
Authors: Torre D. , Speranza F. , Pugliese A. , Fassina G. , Osculati A. , Perversi L. , Banfi M. G. , Airoldi M.
Published: July 2002

To investigate effect of MMLA, an inhibitor of nitric oxide (NO) production, on regulation of inflammatory responses to Bordetella pertussis infection, mice were infected intranasally, and treated with various concentrations of MMLA. Ten days after infection, mice treated with MMLA at dosage of 100 mg/kg, given intraperitoneally in a single dose or for 5 consecutive days, showed at histopathologic examination, a significant decrease of intensity of inflammation (scores, 0.6±0.2 and 0.9±0.5 respectively). A decrease of cellular accumulation of neutrophils and lymphocytes in the bronchoalveolar lavage (BAL) fluid was observed in infected mice treated with MMLA, especially at dosage of 10 mg/kg, given in a single dose intraperitoneally. In addition, BP‐infected mice treated with MMLA (100 mg/kg, intraperitoneally) for 5 consecutive days showed higher mortality rate than untreated mice infected with B. pertussis, and the number of B. pertussis in lungs of mice treated with MMLA was significantly increased. However, MMLA treatment of infected mice had some effect on levels of IFN‐γ and nitrite/nitrate (end‐stable products of NO) in the BAL fluid. This study indicates that NO may play a role either as microbiocidal agent or as a modulator of immune regulation, inasmuch as it may upregulate tissue inflammatory response to B.pertussis.
Topical Application of Tacrolimus Ointment Did Not Alter the Cutaneous Pigmentation of Yucatan Micropigs
Authors: Bekersky Ihor , Boswell Garry , Ohara Kaname , Kuroda Yoshio , Sambuco Christopher
Published: July 2002

Tacrolimus is a potent immunosuppressant marketed for the prevention of rejection in liver and kidney transplantation. Topical tacrolimus has been shown to be effective in the treatment of atopic dermatitis, a disease with an immunologic basis, and is currently being developed for this indication. The objective of the current study was to determine whether repeated topical application of tacrolimus ointment could result in hypopigmentation at the application site(s). Each of 10 Yucatan miniature pigs received topical application of 0.03%, 0.1%, and 0.3% tacrolimus ointment, positive control (a combination of 2% 4-hydroxyanisole and 0.01% all-trans-retinoic acid, 4-HAArRA), and a negative control (vehicle placebo) to five test sites (approximately 12.5 cm2) along either side of the dorsal midline (10 sites per animal). Tacrolimus and controls were randomly assigned to the test site on each animal and were applied unoccluded twice daily for 8 weeks at a dose volume of 0.1 ml per site for tacrolimus and vehicle control and 0.025 ml per site for the positive control. Topical application of tacrolimus ointment in concentrations of up to 0.3% for 8 weeks had no effect on the cutaneous pigmentation of Yucatan miniature swine. In contrast, the application of 4-HAArRA produced a statistically significant (P<.05) induction of hypopigmentation from week 4 through the end of the study application.
Recent Progress in the Management of Retroperitoneal Sarcoma
Authors: Cheifetz Rona , Catton Charles N. , Kandel Rita , O′Sullivan Brian , Couture Jean , Swallow Carol J.
Published: July 2002

Retroperitoneal sarcomas (RPS) are rare tumours that typically present late and carry a poor prognosis even following grossly complete resection. In an attempt to improve the outlook for patients with RPS, sarcoma specialists have employed various adjuvant therapies, including extermal beam radiation, intraoperative radiation, brachyradiation and systemic chemotherapy. This article reviews the presentation and prognosis of RPS, and focuses on the results of new treatment strategies compared with conventional management.

A Medline search of the English literature was performed to identify all retrospective and prospective reports relating to the management of adult RPS published since 1980. Series that did not analyse RPS separately from other intra‐abdominal or extra‐abdominal sarcomas or other malignancies were excluded, and information on investigation, presentation, prognostic
factors, treatment and outcome was extracted from the remaining reports. Survival and local control data were collected from reports that contained at least 30 cases of RPS (n = 31).

While surgical resection remains the cornerstone of treatment for RPS, the majority of patients will relapse and die from sarcoma within 5 years of resection. Adjuvant radiation may improve these results, but further trials are required to definitively demonstrate its benefit. Possible reasons for the failure of conventional treatment are discussed, and alternative strategies designed to overcome these obstacles are presented.
Post‐Operative Radiotherapy of the Rhabdomyosarcoma R1H of the Rat
Authors: Carl Ulrich M. , Sminia Peter , Bahnsen Jens , Fröschle Günter , Omniczynski Maria , Wolf Lothar , Krüger Uwe , Hartmann K. Axel , Beck-Bornholdt Hans-Peter
Published: July 2002

Purpose. Post‐operative radiotherapy (RT) is routinely applied in the treatment of several human tumours. The aim of
the present study was to investigate the value of post‐operative RT in a rat model.

Methods. Experiments were performed using the rhabdomyosarcoma R1H of the WAG/Rij rat. Animals were randomized
to different treatment schedules: surgery, RT or a combination of both. Tumours were excised at different sizes (0.1–4.5 g)
aiming for complete macroscopic resection. RT (60 Gy in 30 daily fractions over 6 weeks) was applied either primarily or to
the former turnout site from the third post‐operative day. Tumour growth delay, time to recurrence and local tumour control
were used as endpoints.

Results. Pre‐operative tumour size determined the time and rate of recurrence. The larger the tumour,
the shorter the time to relapse and the higher the recurrence rate. The 50% local control rate (LCR50) for surgery was found in tumours
with a mass of 0.8 g. For post‐operative RT a LCR50 was achieved for tumours with a mass of 1.1 g. For larger turnouts
(> 1.1 g), however, the rate and time course of relapse were similar for both the group receiving RT alone and the group
receiving post‐operative RT.

Discussion. In this model the tumour mass at excision governs the prognosis. Relatively small R1H turnouts may recur
despite complete macroscopical resection. With regard to the LCR, the outcome for larger tumours is improved with
post‐operative RT (60 Gy/6 weeks) than compared with surgery alone. The factor is 1.3. Within a certain range of tumour
sizes, combined treatment (surgery + RT) can improve the outcome considerably.
IL‐1β Increases uPA and uPA Receptor Expression in Human Gingival Fibroblasts
Authors: Ogura Naomi , Tobe Makiko , Tamaki Hiroyuki , Nagura Hideaki , Abiko Yoshimitsu
Published: July 2002

The binding of urokinase‐type plasminogen activator (uPA) to its receptor (uPAR) in various cell types has been proposed as an important feature of many cellular processes requiring extracellular proteolysis, cell adhesion, motility, and invasion. uPAR attaches to the cell surface with a glycosylphophatidylinositol (GPI) anchor, and serves to localize and accelerate the proteolysis cascade. In this study, we examined both uPA and uPAR levels in human gingival fibroblasts treated with an inflammatory cytokine, interleukin‐1 β(IL‐1 β). PA activity in the cell lysate was increased by treatment with IL‐1 β. Further, PA activity released by phosphatidylinositol‐specific phospholipase C, which detaches the GPI anchor, was also increased by IL‐1 β. The activity was inhibited by amiloride, a specific inhibitor of uPA. In addition, IL‐1 βincreased the protein and mRNA levels of both uPA and uPAR in gingival fibroblasts. These findings suggest that the enhancement of uPA and uPAR levels by IL‐1 βmay play an important role in the progression of periodontal diseases through pericellular proteolysis, and subsequent cellular behavior.
cDNA Cloning of the Basement Membrane Chondroitin Sulfate Proteoglycan Core Protein, Bamacan: A Five Domain Structure Including Coiled-Coil Motifs
Authors: Wu Rong-Rong , Couchman John R.
Published: July 2002

Basement membranes contain several proteoglycans, and those bearing heparan sulfate glycosaminoglycans such as perlecan and agrin usually predominate. Most mammalian basement membranes also contain chondroitin sulfate, and a core protein, bamacan, has been partially characterized. We have now obtained cDNA clones encoding the entire bamacan core protein of Mr = 138 kD, which reveal a five domain, head-rod-tail configuration. The head and tail are potentially globular, while the central large rod probably forms coiled-coil structures, with one large central and several very short interruptions. This molecular architecture is novel for an extracellular matrix molecule, but it resembles that of a group of intracellular proteins, including some proposed to stabilize the mitotic chromosome scaffold. We have previously proposed a similar stabilizing role for bamacan in the basement membrane matrix. The protein sequence has low overall homology, apart from very small NH2- and COOH-terminal motifs.

At the junctions between the distal globular domains and the coiled-coil regions lie glycosylation sites, with up to three N-linked oligosaccharides and probably three chondroitin chains. Three other Ser-Gly dipeptides are unfavorable for substitution. Fusion protein antibodies stained basement membranes in a pattern commensurate with bamacan, and they also Western blotted bamacan core protein from rat L2 cell cultures. The antibodies could also specifically immunoprecipitate an in vitro transcription/translation product from a full-length bamacan cDNA. The unusual structure of this proteoglycan is indicative of specific functional roles in basement membrane physiology, commensurate with its distinct expression in development and changes in disease models.
A Cytosolic Serine Endopeptidase from Trypanosoma cruzi Is Required for the Generation of Ca2+ Signaling in Mammalian Cells
Authors: Burleigh Barbara A. , Caler Elisabet V. , Webster Paul , Andrews Norma W.
Published: July 2002

An early event in the Trypanosoma cruzi cell invasion process, the recruitment of host lysosomes, led us to investigate the involvement of signal transduction. Infective trypomastigotes were found to contain a soluble Ca2+-signaling activity for mammalian cells that is sensitive to protease inhibitors. Inhibitor and substrate utilization profiles were used to purify a candidate peptidase for involvement in this process, from which we isolated a full-length cDNA clone. The sequence revealed a novel enzyme, denominated T. cruzi oligopeptidase B, which is homologous to members of the prolyl oligopeptidase family of serine hydrolases, known to participate in the maturation of biologically active peptides. The T. cruzi oligopeptidase B was expressed as a fully active product in Escherichia coli, and antibodies to the recombinant enzyme inhibited both peptidase activity and Ca2+ signaling induced in normal rat kidney cells by trypomastigote extracts. Our data suggest that the T. cruzi oligopeptidase B participates in processing events in the cytoplasm of the parasites, generating a factor with Ca2+-signaling activity for mammalian cells.
Threshold Levels of Fluid Shear Promote Leukocyte Adhesion through Selectins (CD62L,P,E)
Authors: Lawrence Michael B. , Kansas Geoffrey S. , Kunkel Eric J. , Ley Klaus
Published: July 2002

Leukocyte adhesion through L-selectin to peripheral node addressin (PNAd, also known as MECA-79 antigen), an L-selectin ligand expressed on high endothelial venules, has been shown to require a minimum level of fluid shear stress to sustain rolling interactions (Finger, E.B., K.D. Puri, R. Alon, M.B. Lawrence, V.H. von Andrian, and T.A. Springer. 1996. Nature (Lond.). 379:266–269). Here, we show that fluid shear above a threshold of 0.5 dyn/cm2 wall shear stress significantly enhances HL-60 myelocyte rolling on P- and E-selectin at site densities of 200/μm2 and below. In addition, gravitational force is sufficient to detach HL60 cells from P- and E-selectin substrates in the absence, but not in the presence, of flow. It appears that fluid shear–induced torque is critical for the maintenance of leukocyte rolling. K562 cells transfected with P-selectin glycoprotein ligand-1, a ligand for P-selectin, showed a similar reduction in rolling on P-selectin as the wall shear stress was lowered below 0.5 dyn/cm2. Similarly, 300.19 cells transfected with L-selectin failed to roll on PNAd below this level of wall shear stress, indicating that the requirement for minimum levels of shear force is not cell type specific. Rolling of leukocytes mediated by the selectins could be reinitiated within seconds by increasing the level of wall shear stress, suggesting that fluid shear did not modulate receptor avidity. Intravital microscopy of cremaster muscle venules indicated that the leukocyte rolling flux fraction was reduced at blood centerline velocities less than 1 mm/s in a model in which rolling is mediated by L- and P-selectin. Similar observations were made in L-selectin–deficient mice in which leukocyte rolling is entirely P-selectin dependent. Leukocyte adhesion through all three selectins appears to be significantly enhanced by a threshold level of fluid shear stress.
Targeting Determinants and Proposed Evolutionary Basis for the Sec and the Delta pH Protein Transport Systems in Chloroplast Thylakoid Membranes
Authors: Henry Ralph , Carrigan Matthew , McCaffery Michael , Ma Xianyue , Cline Kenneth
Published: July 2002

Transport of proteins to the thylakoid lumen is accomplished by two precursor-specific pathways, the Sec and the unique Delta pH transport systems. Pathway selection is specified by transient lumen-targeting domains (LTDs) on precursor proteins. Here, chimeric and mutant LTDs were used to identify elements responsible for targeting specificity. The results showed that: (a) minimal signal peptide motifs consisting of charged N, hydrophobic H, and cleavage C domains were both necessary and sufficient for pathway-specific targeting; (b) exclusive targeting to the Delta pH pathway requires a twin arginine in the N domain and an H domain that is incompatible with the Sec pathway; (c) exclusive targeting to the Sec pathway is achieved by an N domain that lacks the twin arginine, although the twin arginine was completely compatible with the Sec system. A dual-targeting signal peptide, constructed by combining Delta pH and Sec domains, was used to simultaneously compare the transport capability of both pathways when confronted with different passenger proteins. Whereas Sec passengers were efficiently transported by both pathways, Delta pH passengers were arrested in translocation on the Sec pathway. This finding suggests that the Delta pH mechanism evolved to accommodate transport of proteins incompatible with the thylakoid Sec machinery.
Isoforms of Ankyrin-3 That Lack the NH2-terminal Repeats Associate with Mouse Macrophage Lysosomes
Authors: Hoock Thomas C. , Peters Luanne L. , Lux Samuel E.
Published: July 2002

We have recently cloned and characterized ankyrin-3 (also called ankyrinG), a new ankyrin that is widely distributed, especially in epithelial tissues, muscle, and neuronal axons (Peters, L.L., K.M. John, F.M. Lu, E.M. Eicher, A. Higgins, M. Yialamas, L.C. Turtzo, A.J. Otsuka, and S.E. Lux. 1995. J. Cell Biol. 130: 313–330). Here we show that in mouse macrophages, ankyrin-3 is expressed exclusively as two small isoforms (120 and 100 kD) that lack the NH2-terminal repeats. Sequence analysis of isolated Ank3 cDNA clones, obtained by reverse transcription and amplification of mouse macrophage RNA (GenBank Nos. U89274 and U89275), reveals spectrin-binding and regulatory domains identical to those in kidney ankyrin-3 (GenBank No. L40631) preceded by a 29–amino acid segment of the membrane (“repeat”) domain, beginning near the end of the last repeat. Antibodies specific for the regulatory and spectrin-binding domains of ankyrin-3 localize the protein to the surface of intracellular vesicles throughout the macrophage cytoplasm. It is not found on the plasma membrane. Also, epitope-tagged mouse macrophage ankyrin-3, transiently expressed in COS cells, associates with intracellular, not plasma, membranes. In contrast, ankyrin-1 (erythrocyte ankyrin, ankyrinR), which is also expressed in mouse macrophages, is located exclusively on the plasma membrane. The ankyrin-3–positive vesicles appear dark on phasecontrast microscopy. Two observations suggest that they are lysosomes. First, they are a late compartment in the endocytic pathway. They are only accessible to a fluorescent endocytic tracer (FITC-dextran) after a 24-h incubation, at which time all of the FITC-dextran– containing vesicles contain ankyrin-3 and vice versa. Second, the ankyrin-3–positive vesicles contain lysosomal-associated membrane glycoprotein (LAMP-1), a recognized lysosomal marker. This is the first evidence for the association of an ankyrin with lysosomes and is an example of two ankyrins present in the same cell that segregate to different locations.
A Mitosis-specific Phosphorylation of the Gap Junction Protein Connexin43 in Human Vascular Cells: Biochemical Characterization and Localization
Authors: Xie Han-qing , Laird Dale W. , Chang Tsg-Hui , Hu Valerie W.
Published: July 2002

Western blotting studies revealed that connexin43 (Cx43), one of the major gap junction proteins in human vascular endothelial cells, is posttranslationally modified during mitosis. This mitosis-specific modification results in a Cx43 species that migrates as a single protein band and was designated Cx43m. Cx43m was shown to be the result of additional Ser/Thr phosphorylation as indicated by: (a) the increased gel mobility induced by both alkaline phosphatase and the Ser/ Thr-specific protein phosphatase-2A (PP2A) and (b) the removal of virtually all 32Pi from Cx43m by PP2A. Immunofluorescent confocal microscopy of mitotic cells revealed that Cx43 is intracellularly located, while in nonmitotic cells Cx43 is located at regions of cell–cell contact. Dye coupling studies revealed that mitotic endothelial cells were uncoupled from each other and from nonmitotic cells. After cytokinesis, sister cells resumed cell coupling independent of de novo protein synthesis. The mitosis-specific phosphorylation of Cx43 correlates with the transient loss of gap junction intercellular communication and redistribution of Cx43, suggesting that a protein kinase that regulates gap junctions is active in M-phase.