bluuebunny/crossref_metadata_2025 huggingface dataset:
Discrimination of oestrous status in giant pandas (Ailuropoda melanoleuca) via chemical cues in urine
Authors: Swaisgood R. R. , Lindburg D. G. , Zhang H.
Published: July 2002

Giant pandas Ailuropoda melanoleuca are a solitary, seasonally mono‐oestrous species, and as such are likely to rely heavily on chemical signals to co‐ordinate mating efforts. Perhaps the most crucial chemosensory task facing male pandas is the assessment of female reproductive condition on the basis of chemical cues. To test for this ability, eight male and 10 female giant pandas were presented with urine from oestrous and non‐oestrous females. Males but not females displayed a significant investigatory preference for oestrous over non‐oestrous female urine, as indicated by sniffing and flehmen. Males also investigated, displayed flehmen, and licked female urine more than did females. These findings indicate that female urine contains chemical cues permitting discrimination of oestrous status, and probably serves a reproductive advertisement function. The prevalence of flehmen and licking by males suggests that males may rely on the vomeronasal organ to assess female reproductive condition. Female odours may play an important role in the reproductive behaviour of male giant pandas and, therefore, may have implications for captive‐breeding programmes.
Karyotype and X–Y chromosome pairing in the Sikkim vole (Microtus (Neodon) sikimensis)
Authors: Mekada Kazuyuki , Koyasu Kazuhiro , Harada Masashi , Narita Yuichi , Shrestha Krishna C. , Oda Sen‐Ichi
Published: July 2002

Pairing of the X and Y chromosomes during meiotic prophase and the G‐ and C‐banding patterns were analysed in Microtus sikimensis. The karyotype of M. sikimensis from central Nepal, has 2n= 48 and FNa = 56. It has been derived from the all single‐armed karyotype of M. carruthersi through a unique course of evolution. There is a synapsis between the X and Y chromosomes at pachytene and end‐to‐end association from diakinesis to metaphase I. Microtus sikimensis has no close karyological or morphological relationship to the other synaptic species studied so far. These data confirm that several species or lineages with synaptic condition exist in the genus Microtus, and that the ancestor of Microtus had synaptic sex chromosomes.
E-language learning for the airline industry
Authors: GIMENO-SANZ ANA
Published: July 2002

The acquisition of foreign languages for specific purposes through
e-learning strategies is one of the areas less researched and developed in the field of CALL.
However, as a member of a recently completed Leonardo da Vinci Project, devoted to vocational
training, the author will report on the findings of the Airline Talk project. The goals of the
project were to supply materials which would promote continuing learning amongst airline staff in
three target languages: English, German and Spanish. This paper focuses on the development,
structure and language content of the Spanish courseware, ¡Bienvenido a bordo!, designed at
the Universidad Politécnica de Valencia. The author discusses the process carried out by
the project partners in order to satisfy learner demands in terms of learner orientation,
process orientation and learner autonomy to achieve language competence through the development of
communicative skills and interactivity. Issues that are described include: needs analysis,
courseware specifications, exercise template design, graphical user interface and learner motivation.
The results arising from the evaluation carried out at two different partner institutions will
conclude the discussion.
Web-enhanced approaches to the teaching of linguistic variation in French
Authors: McBRIDE NICOLE
Published: July 2002

This paper considers the progressive integration of a web resource in a
language degree option centred on language varieties and linguistic variation in French.
It focuses on the features introduced by the web environment and how they can enhance the
learning experience of the students. It addresses the issue of the language used for teaching
and learning with diverse groups of students including a high proportion of non-native speakers
of English, varying levels of linguistic competence and different stages of study. The paper also
examines web-based facilities supporting interaction and collaboration among members of the
learning group (some teacher moderated). The evaluation is based on the results of a survey
conducted with the group of students who took part in the pilot implementation in 2000–01 and
on module feedback. The last section in the paper points to ways of integrating further the
technology within the delivery and the assessment of the module, and of increasing student support
and interaction.
Introduction
Authors: Corradini Andrea , Ehrig Hartmut , Rozenberg Grzegorz , Taentzer Gabriele
Published: July 2002

This special issue of Mathematical Structures in Computer Science is devoted to the theory
and applications of graph transformations. This research area dates back to the early
seventies and is based on mathematical techniques from graph theory, algebra, logic
and category theory. The theory of graph transformations has become attractive as a
modelling and programming paradigm for complex graphical structures in a large variety
of areas in computer science and for applications to other fields. During the Joint
APPLIGRAPH/GETGRATS Workshop on Graph Transformation Systems (GRATRA
2000) – a satellite of ETAPS 2000 in Berlin – 35 lectures were presented by participants
from all over the world. The authors of presentations that stressed the theoretical point of
view were invited to submit a full version of their presentation to Mathematical Structures
in Computer Science. After a careful refereeing process nine papers have been accepted,
seven of which are included in this special issue.

The paper by Ehrenfeucht, Petre, Prescott and Rozenberg connects the important new
area of DNA computing in vivo to our area of graph transformation. An application
of hypergraph constructions to the static analysis of concurrent systems is presented
by König, and normal forms for context-free node rewriting hypergraph grammars by
Klempien-Hinrichs. The construction of pushout complements for ‘partly total algebras’
generalising attributed graphs is presented by Burmeister, Llabrés and Roselló. Finally,
Courcelle and Makowsky present operations on relational structures (generalising different
kinds of graphs and hypergraphs) that are compatible with monadic second order logic.
Four other accepted papers could not be included in this special issue because of space
limitations.

They will appear in regular issues of MSCS:

— R. Heckel, M. Llabrés, H. Ehrig and F. Orejas: Concurrency and loose semantics of
open graph transformation systems.

— L. Helouet, C. Jard and B. Caillaud: An event structure based semantics for high-level
message sequence charts.

— J. Larossa and G. Valiente: Constraint satisfaction algorithms for graph pattern
matching.

— N. Verlinden and D. Janssens: Algebraic properties for Local Action Systems.

We are most grateful to all the referees of these papers, to Olga Runge and Claudia
Ermel for technical support, and to Giuseppe Longo and Cambridge University Press for
fruitful cooperation in editing this special issue.
Reply to De Grand: But Did Reform Fail?
Authors: Corner Paul
Published: July 2002

I am indebted to Alexander De Grand for taking the time and trouble to comment on my article, although I am sorry that he seems to have read it principally as an attack on Giolitti and Giolittian policies. While I can see that it is possible to read this into the paper, it was certainly not my intention to lay the responsibility for the development of Fascism at Giolitti's door. My concern was rather to seek to identify some of the reasons for the dramatic clash between left and right in 1920 and 1921 which led to the affirmation of Fascism; it was in this light that I attempted to assess the Giolittian period, which has always seemed to me the great moment of democratic possibilities between one form of repressive government and another. I agree, of course, that the great radicalising and polarising event in Italy was the First World War. My point – put very simply – was that the experience of the war might have been much less devastating for Italy if the political situation in 1914–15 had not already been characterised by profound lacerations within Italian society. To put it another way, I was interested in seeing why, as De Grand himself says, ‘the great hopes for reform that marked that [Giolittian] period gave rise to little structural reform’ and in assessing the consequences of that lack of reform for the subsequent period. The central question of the article, therefore, is that of Fascism as breach or continuity rather than the culpability of Giolitti.
New Friends – Difficult Friendships: Germany and its Western Neighbours in the Postwar Era
Authors: Schönwald Matthias
Published: July 2002

Daniel Gossel, Briten, Deutsche und Europa. Die Deutsche Frage in der britischen Außenpolitik 1945–1962 (Stuttgart: Steiner, 1999), 259pp., DM 88.00, ISBN 3-515-07159-8. Klaus Larres, ed., Uneasy Allies: British-German Relations and European Integration Since 1945 (Oxford:Oxford University Press, 2000) 344pp., $85.00, ISBN 0-19-829383-6. Werner Rouget, Schwierige Nachbarschaft am Rhein: Frankreich–Deutschland, ed. Ernst Weisenfeld and Joachim Bitterlich (Bonn: Bouvier, 1998), 160pp., DM 29.80 ISBN 3-416-02762-0. Friso Wielenga, Vom Feind zum Partner. Die Niederlande und Deutschland seit 1945. (Münster: Agenda Verlag, 2000), 522pp., DM 59.80, ISBN 3-896880721. Robert Bohn, Jürgen Elvert and Karl Christian Lammers, eds., Deutsch-skandinavische Beziehungen nach 1945 (Stuttgart: Steiner, 1998), 280pp., DM 96.00, ISBN 3-515-07320-5. Birgit Aschmann, ‘Treue Freunde. . .’? Westdeutschland und Spanien 1945–1963 (Stuttgart: Steiner, 1999), 502pp., DM 188.00, ISBN 3-515-07579-8. Guido Müller, ed., Deutschland und der Westen: Internationale Beziehungen im 20. Jahrhundert, Festschrift für Klaus Schwabe. (Stuttgart: Steiner, 1998), 381pp., DM 138.00, ISBN 3-515-07251-9. Gottfried Niedhart, Detlef Junker and Michael W. Richter, eds., Deutschland in Europa. Nationale Interessen und internationale Ordnung im 20.Jahrhundert. (Mannheim: Palatium, 1997), 397pp., DM 98.00, ISBN 3-920671-29-5.
Identification of distinct K+ channels in mouse spermatogenic cells and sperm
Authors: Felix Ricardo , Serrano Carmen J. , Treviño Claudio L. , Muñoz-Garay Carlos , Bravo Alejandra , Navarro América , Pacheco Judith , Tsutsumi Victor , Darszon Alberto
Published: July 2002

Potassium (K+) channels are believed to regulate mammalian sperm acquisition of fertilising capacity. However, the molecular identity of these proteins in sperm has not been elucidated. In this report, using immunoconfocal and electron microscopy we show that a minimum of four different classes of K+ channels (Kv1.1, Kv1.2, Kv3.1 and GIRK1) are present and regionally distributed over the surface of mouse epididymal sperm. In addition, the use of reverse transcription polymerase chain reaction on RNA from mouse spermatogenic cells allowed the amplification of multiple transcripts corresponding to the channels identified by immunocytochemistry. Consistent with this, whole-cell patch-clamp recordings showed the expression of at least two different outwardly rectifying K+ currents in spermatogenic cells.
New worlds outside the Solar System
Authors: SCHNEIDER JEAN
Published: July 2002

The discoveries of the first planets around a pulsar and around a star similar to our Sun have opened, after centuries of speculation, a new era in astronomy. The quest for other worlds has become reality. In this paper, the motivations for this search are presented. A comparative review is made of different techniques for their detection, with special attention paid to the planet parameters that can be derived from each method. The first discoveries and their astrophysical consequences are analysed. New questions and future projects beyond these first discoveries are reviewed, with special attention given to the search for life in extrasolar planetary systems. Finally, worldwide activities and the role of Europe are described.
Optical and infrared astronomy in the 21st century – the continuing revolution
Authors: CESARSKY CATHERINE , WEST RICHARD
Published: July 2002

For some decades, astronomy and astrophysics have undergone a technological and conceptual revolution. Supported by ever more powerful telescopes and instruments on the ground and in space, the volume and quality of new insights is incredible, both in terms of physical understanding of individual celestial objects and the grand evolutionary scheme. New and powerful observational facilities such as the ESO Very Large Telescope (VLT) are opening new horizons, from the nearby solar system to the corners of the Milky Way galaxy in which we live and, not least, towards the vast expanses in time and space of the remote and early Universe. The next generation of ultra-sensitive optical-infrared telescopes such as Herschel and ALMA will be ready within this decade and concepts are being elaborated for the construction of super-giant telescopes like the 100 m optical/IR OWL, the ‘Overwhelmingly Large telescope’. With these impressive developments, and in a true spirit of exploration, astronomers can now look forward to great research opportunities, in a resounding manifestation of the continuous drive towards a better understanding of our cosmic surroundings and of our own origins, so characteristic for enlightened humankind of every age.
Estimation of local near‐surface wind conditions –a comparison of WASP and regression based techniques
Authors: Achberger C , Ekström M , Bärring L
Published: July 2002

This study compares the performance of different models used to assess the local wind near‐surface conditions at an agricultural site in Scania, southern Sweden. The methods are: (a) the WASP model (Wind Analysis and Application Program), (b) separate linear regressions of the two wind vector components, (c) a regression model based on vector correlation, and (d) linear regression of scalar wind. Each method was tested with three different data sets over nine months: wind measurements from the nearby Sturup airport SYNOP station, 10 m surface wind and surface geostrophic wind produced by the operational Swedish Mesoscale Analysis system (Mesan). The wind climate estimations were compared with observed winds at the field site, with respect to mean wind speed, wind direction, wind speed frequency distribution and the relative frequency of winds above 6 m s−1. All models performed reasonably well with data from Sturup and Mesan surface wind, but gave less reliable results with the Mesan geostrophic data. The estimated frequency of winds above 6 m s−1 was in general lower than the observed frequency. Overall, best results were obtained with WASP in combination with measurements from Sturup. Copyright © 2002 Royal Meteorological Society
Using the GANDOLF system as a tool to aid the forecasting of lightning strikes
Authors: Soul K M , Archibald E J , Hardaker P J , Hounsell A
Published: July 2002

In partnership with the Environment Agency, the Met Office has developed a short‐period thunderstorm forecasting system based on a conceptual life‐cycle model of convective cells. Although originally intended to provide forecasts of intense precipitation, the model provides a mechanism for predicting other forms of severe weather phenomena associated with thunderstorms. This paper investigates the possibility of using GANDOLF to generate a forecast of lightning risk. Model life‐cycle stages are compared against observed lightning locations in order to derive a basis for forecasting risk, and this was then tested on data for three different events. The results from this initial investigation seem promising and suggest this type of approach can be effective. Copyright © 2002 Royal Meteorological Society
Do public works decrease farmers' soil degradation? Labour income and the use of fertilisers in India's semi-arid tropics
Authors: Berg Marrit van den
Published: July 2002

This paper investigates the possibility of using public works to stimulate farmers' fertiliser use in India's SAT. Inadequate replenishment of removed nutrients and organic matter has reduced fertility and increased erosion rates. Fertiliser use, along with other complementary measures, can help reverse this process, which ultimately leads to poverty, hunger, and further environmental degradation. In a high-risk environment like India's SAT, there may be a strong relation between off-farm income and smallholder fertiliser use. Farmers can use the main source of off-farm income, wage income, to manage risk as well as to finance inputs. Consequently, the introduction of public works programmes in areas with high dry-season unemployment may affect fertiliser use. This study confirms the relevance of risk for decisions regarding fertiliser use in two Indian villages. Nevertheless, governments cannot use employment policies to stimulate fertiliser use. Public works even decrease fertiliser use in the survey setting.
Working with the given
Authors: Blundell Jones Peter
Published: July 2002

In this essay the author suggests that although the resulting
architectural style may be ambiguous, a dialogue with an existing
place can be creative, rewarding and appropriate. Just how this
might be so is demonstrated in the following article, ‘New meanings
from old buildings’, on p.312—331.
The artist's life: notes on looking at C.R. Mackintosh
Authors: McKean John
Published: July 2002

Last time I was in Glasgow, I met the
architect Isi Metzstein in the street
and his first words to me were: ‘It's
you! I don't know how you can
show your face here!’ My book on
Mackintosh had just been
published, and local papers were
carrying headlines like
‘MACKINTOSH AUTISTIC!’
Conditions for the voicing of Old English fricatives, II: morphology and syllable structure
Authors: Fulk R. D.
Published: July 2002

Old English fricatives at points of morpheme juncture are studied to determine whether they conform to the rule of voicing between voiced sounds that applies morpheme-internally. Should we expect a voiced or a voiceless fricative in words like OE heorð-weorod, Wulfweard, and stīðlīce? The evidence examined regards chiefly compounds and quasi-compounds (the latter comprising both forms bearing clear derivational affixes and ‘obscured’ compounds, those in which the deuterotheme has lost its lexical independence), though a small amount of evidence in regard to voicing before inflectional suffixes is considered. Evidence is derived from place-names, personal names, and common nouns, on the basis of Modern English standard pronunciation, assimilatory changes in Old English, modern dialect forms, post-Conquest and nonstandard Old English spellings, and analogous conditioning for the loss of OE /x/. A considerable preponderance of the evidence indicates that in compounds as well as in quasi-compounds, fricatives were voiced at the end of the prototheme when a voiced sound followed, but not a voiceless one. It follows from the evidence that there was no general devoicing of fricatives in syllable-final position in Old English, despite Anglo-Saxon scribes' use of  for etymological [Γ] in occasional spellings like  and . Old English spellings of this kind need be taken to imply nothing more than a tendency for  and  to be used interchangeably in noninitial positions, due to the noncontrastive distribution of the sounds they represent everywhere except morpheme-initially. Rare early Middle English spellings of this kind may or may not have a phonological basis, but they cannot plausibly be taken to evidence a phonological process affecting /v, ð, z/.
How many protected minke whales are sold in Japan and Korea? A census by microsatellite DNA profiling
Authors: Dalebout Merel L. , Lento Gina M. , Cipriano Frank , Funahashi Naoko , Baker C. Scott
Published: July 2002

Products from the protected East Sea/Sea of Japan (‘J’ stock) minke whales (Balaenoptera acuturostrata) are sold widely on the commercial markets of Japan and Korea despite the protection of this stock since 1986. To determine the minimum number of individual whales for sale, genotypes from six microsatellite loci were used to profile North Pacific (NP) minke whale products purchased on these markets between December 1997 and October 1999. Genotype differences showed that 99 NP minke whale products from the Japanese market represented 86 unique individuals. Of these, 33.7% were of likely J‐stock origin based on mitochondrial (mt) DNA haplotypes. In Korea, genotyping showed that 23 NP minke products from March 1999 represented 18 individuals, and 19 products from October 1999 represented 16 individuals. No matches were found between the two sampling periods, giving a total of 34 unique individuals. A frequency‐of‐capture model suggests that 98 minke whales were present on the Korean market over the two brief sampling periods. No genotype matches were confirmed between the two countries, indicating that undocumented exploitation of this depleted stock must be additive, and greater than previously assumed.
A phylogenetic supertree of the bats (Mammalia: Chiroptera)
Authors: JONES KATE E. , PURVIS ANDY , MacLARNON ANN , BININDA‐EMONDS OLAF R. P. , SIMMONS NANCY B.
Published: July 2002

We present the first estimate of the phylogenetic relationships among all 916 extant and nine recently extinct species of bats (Mammalia: Chiroptera), a group that accounts for almost one‐quarter of extant mammalian diversity. This phylogeny was derived by combining 105 estimates of bat phylogenetic relationships published since 1970 using the supertree construction technique of Matrix Representation with Parsimony (MRP). Despite the explosive growth in the number of phylogenetic studies of bats since 1990, phylogenetic relationships in the order have been studied non‐randomly. For example, over one‐third of all bat systematic studies to date have focused on relationships within Phyllostomidae, whereas relationships within clades such as Kerivoulinae and Murinae have never been studied using cladistic methods. Resolution in the supertree similarly differs among clades: overall resolution is poor (46.4% of a fully bifurcating solution) but reaches 100% in some groups (e.g. relationships within Mormoopidae). The supertree analysis does not support a recent proposal that Microchiroptera is paraphyletic with respect to Megachiroptera, as the majority of source topologies support microbat monophyly. Although it is not a substitute for comprehensive phylogenetic analyses of primary molecular and morphological data, the bat supertree provides a useful tool for future phylogenetic comparative and macroevolutionary studies. Additionally, it identifies clades that have been little studied, highlights groups within which relationships are controversial, and like all phylogenetic studies, provides preliminary hypotheses that can form starting points for future phylogenetic studies of bats.
State Rebuilding, Popular Protest and Collective Action in China
Authors: Zheng Yongnian
Published: July 2002

Reforms in post-Mao China have led to the rise of social movements and collective action. The Falun Gong movement, a semi-religious movement, in particular has caught worldwide attention. Indeed, social protests have become a norm in China.
Social Exclusion in a Consumer Society: A Study of Five Rural Communities
Authors: Williams Colin C.
Published: July 2002

The starting point of this paper is that the shift from a producer to a consumer society necessitates that social exclusion is investigated not only in relation to production but also consumption. To do this, case study evidence from interviews with 350 households in rural England is reported. This reveals that people define themselves as excluded from mainstream consumption not only when they are unable to acquire goods but also when they rely on informal and second-hand channels (e.g., car boot sales, second-hand shops) to obtain them. Social exclusion in consumer society is thus found to relate to not only non-possession but also exclusion from mainstream modes of goods acquisition.
Steady state sediment transport through an area of net deposition: Multisize class solutions
Authors: Sander Graham C. , Hairsine Peter B. , Beuselinck Laurent , Govers Gerard
Published: July 2002

In this paper, analytical solutions are developed for the multisize class equations derived by Hairsine et al. [2002] for steady state sediment transport over areas of net deposition. Under conditions of a constant flow velocity we are able to decouple the equations and reduce the solution to a straightforward integral for the largest size class. All other size class solutions are then given by an algebraic equation. For practical applications we also derive simple but accurate approximations to this integral, resulting in solutions for all size classes being known explicitly as a function of distance. The effect of infiltration within a net deposition zone is then considered; however, decoupling of the governing equations is no longer possible as the flow velocities are now spatially dependent, and a full numerical solution is required.
Autoimmune cholangiopathy associated with systemic lupus erythematosus
Authors: Heyman Samuel N. , Spectre Galia , Aamar Suhail , Rubinger Dvora , Pappo Orit , Ackerman Zvi
Published: July 2002

Abstract:  We report a 39‐year‐old female who presented over 11 years with autoimmune cholangiopathy associated with kaleidoscopic manifestations of systemic lupus erythematosus (SLE), including, arthritis, skin changes, pleuritis, diffuse proliferative glomerulonephritis, lymphadenopathy, splenomegaly, hyperglobulinemia, and major depression. While antimitochondrial antibodies (AMA) were absent, antinuclear (ANA) and anti‐DNA antibodies were detected in high titres associated with hypocomplementemia. The patient also had vitamin B12 deficiency and antiphospholipid antibodies. The patient required steroids and repeated courses of cyclophosphamide for the management of lupus nephritis, and ursodeoxycholic acid (ursolite) administration resulted in amelioration of cholestatic laboratory abnormalities. This unusual case report and review of literature illustrate that immune liver disease may be an important clinical manifestation of SLE, especially autoimmune cholangiopathy.
Enhanced expression of endothelial nitric oxide synthase and caveolin‐1 in human cirrhosis
Authors: Yokomori Hiroaki , Oda Masaya , Ogi Mariko , Sakai Kaoru , Ishii Hiromasa
Published: July 2002

Abstract:  Background/aims: Nitric oxide is synthesized in diverse mammalian tissues by a family of calmodulin‐dependent nitric oxide synthases (NOS). Caveolin, the principal structural protein in caveolae, interacts with endothelial NOS (eNOS) leading to enzyme inhibition by a reversible process modulated by Ca++ ‐calmodulin. The aim of the present study was to examine the localizations of eNOS and caveolin‐1 at protein level in normal human liver tissue, and how the expressions are altered in cirrhotic liver.

Methods: Fresh liver specimens were obtained from hepatic surgeries. Normal portions resected from cases of carcinoma metastasized to the liver were used as control specimens, and cirrhotic portions resected from cases of hepatocellular carcinoma with hepatitis C‐related cirrhosis were used as cirrhotic specimens. Anti‐eNOS and anticaveolin‐1 antibodies were used for immunohistochemistry and Western blotting. Immunoelectron microscopy was conducted on ultra thin sections using immunoglobulin–gold combined with silver staining.

Results: Immunohistochemistry revealed that both eNOS and caveolin‐1 were sparsely expressed on hepatic sinusoidal lining in normal liver specimens, and these findings were confirmed by Western blot. Both immunohistochemistry and Western blotting demonstrated over‐expression of eNOS and caveolin‐1 in cirrhotic liver specimens. Morphometric analysis of immunogold particle labeling for eNOS and caveolin‐1 was performed on immunoelectron micrographs. In normal liver tissue, hepatic stellate cells and sinusoidal endothelial cells (SEC) expressed low levels of caveolin‐1, and SEC expressed a very low level of eNOS. In cirrhotic liver, both caveolin‐1 and eNOS expressions were significantly increased by approximately four‐fold on SEC compared to normal liver.

Conclusion: In cirrhotic human liver, marked increase of caveolin‐1 in perisinusoidal cells may promote caveolin‐eNOS binding and reduce the activity of eNOS despite an increased eNOS expression, leading to impaired NO production and increased hepatic microvascular tone.
Quantitative testing of liver function compared to prognostic scores in patients with primary biliary cirrhosis
Authors: Herold Christoph , Ganslmayer Marion , Deynet Christine , Hahn Eckhart G. , Schuppan Detlef
Published: July 2002

Abstract:  Aim: Primary biliary cirrhosis (PBC) is a slowly progressive liver disease which can lead to cirrhosis. We investigated if quantitative tests of liver function (QTLF) and serum levels of a surrogate marker of hepatic fibrogenesis (PIIINP) provide information in addition to established prognostic scores.

Methods: In 34 PBC patients PIIINP, PBC‐relevant parameters, histological staging and QTLF at entry and at 2 years were determined and compared with the Christensen (CPS I, CPS II) and Mayo prognosis score. QTLF included aminopyrine breath test, galactose elimination capacity, sorbitol and indocyanine green clearance.

Results: Bilirubin, serum IgM and PIIINP were elevated at both time points, whereas albumin and prothrombin time remained normal. Clinical findings (ascites, cirrhosis, central cholestasis) and histological staging worsened after 2 years, as did the CPS II. However, QTLF, PIIINP, CPS I and the Mayo score revealed no significant changes.

Conclusions: Only CPS II changed after 2 years, whereas CPS I and the Mayo score remained unaltered. QTLF and PIIINP did not provide any further information on progression of PBC, suggesting that QTLF cannot predict prognosis of PBC patients in a two‐year interval and that CPS II is superior to CPS I and the Mayo score in short‐term studies for PBC.
Enhanced inhibition of L‐type calcium currents by troglitazone in streptozotocin‐induced diabetic rat cardiac ventricular myocytes
Authors: Arikawa Masaya , Takahashi Naohiko , Kira Tetsuya , Hara Masahide , Saikawa Tetsunori , Sakata Toshiie
Published: July 2002

Troglitazone, an insulin‐sensitizing agent shown to improve cardiac function in both experimental animals and patients with diabetes, inhibits voltage‐dependent L‐type Ca2+ currents (ICa,L) in cardiac myocytes, which may underlie its cardioprotective effects. However, inhibition by troglitazone of ICa,L in diabetic cardiac myocytes has not been characterized.

Using whole‐cell voltage‐clamp techniques, ICa,L was measured in ventricular myocytes isolated from 4–6 weeks streptozotocin (STZ)‐induced diabetic rats and age‐matched control rats.

Under control conditions with CsCl internal solution, diabetic myocytes did not differ from control myocytes in membrane capacitance, current density or voltage‐dependent properties of ICa,L.

Troglitazone decreased amplitude of ICa,L in both control and diabetic myocytes in a concentration‐dependent manner. This inhibition was more potent in diabetic than in control myocytes; half‐maximum inhibitory concentrations of troglitazone measured at a holding potential of −50 mV were 4.3 and 9.5 μmol l−1, respectively.

Troglitazone at 5 μmol l−1 did not significantly influence the voltage dependency of steady‐state inactivation or the inactivation time course of ICa,L in either control or diabetic myocytes.

Since troglitazone inhibits ICa,L more effectively in STZ‐induced diabetic ventricular myocytes, this agent may prevent cardiac dysfunction in diabetes.

British Journal of Pharmacology (2002) 136, 803–810. doi:10.1038/sj.bjp.0704757
Functional studies on human Pex7p: subcellular localization and interaction with proteins containing a peroxisome-targeting signal type 2 and other peroxins
Authors: GHYS Karen , FRANSEN Marc , MANNAERTS Guy P. , Van VELDHOVEN Paul P.
Published: July 2002

Pex7p is a WD40-containing protein involved in peroxisomal import of proteins containing an N-terminal peroxisome-targeting signal (PTS2). The interaction of human recombinant Pex7p expressed in different hosts/systems with its PTS2 ligand and other peroxins was analysed using various experimental approaches. Specific binding of human Pex7p to PTS2 could be demonstrated only when Pex7p was formed in vitro by a coupled transcription/translation system or synthesized in vivo in Chinese hamster ovary K1 cells transfected with a construct coding for a Pex7p-green fluorescent protein (GFP) fusion protein. Apparently, no cofactors are required and only monomeric Pex7p binds to PTS2. The interaction is reduced upon cysteine alkylation and is impaired upon truncation of the N-terminus of Pex7p. Interaction of Pex7p with other peroxins could not be demonstrated in bacterial or yeast two-hybrid screens, or in pull-down binding assays. The GFP fusion proteins, tagged at either the N- or C-terminus, were able to restore PTS2 import in rhizomelic chondrodysplasia punctata fibroblasts, and Pex7p-GFP was located both in the lumen of peroxisomes and in the cytosol.
Assembly of the Drosophila 26 S proteasome is accompanied by extensive subunit rearrangements
Authors: KURUCZ Éva , ANDÓ István , SÜMEGI Máté , HÖLZL Harald , KAPELARI Barbara , BAUMEISTER Wolfgang , UDVARDY Andor
Published: July 2002

The subunit contacts in the regulatory complex of the Drosophila 26 S proteasome were studied through the cross-linking of closely spaced subunits of the complex, and analysis of the cross-linking pattern in an immunoblot assay with the use of subunit-specific monoclonal antibodies. The cross-linking pattern of the purified 26 S proteasome exhibits significant differences as compared with that of the purified free regulatory complex. It is shown that the observed differences are due to extensive rearrangement of the subunit contacts accompanying the assembly of the 26 S proteasome from the regulatory complex and the 20S proteasome. Cross-linking studies and electron microscopic examinations revealed that these changes are reversible and follow the assembly or the disassembly of the 26 S proteasome. Although the majority of the changes observed in the subunit contacts affected the hexameric ring of the ATPase subunits, the alterations extended over the whole of the regulatory complex, affecting subunit contacts even in the lid subcomplex. Changes in the subunit contacts, similar to those in the regulatory complex, were detected in the 20S proteasome. These observations indicate that the assembly of the 26 S proteasome is not simply a passive docking of two rigid subcomplexes. In the course of the assembly, the interacting subcomplexes mutually rearrange their structures so as to create the optimal conformation required for the assembly and the proper functioning of the 26S proteasome.
Lipid-restricted recognition of mycobacterial lipoglycans by human pulmonary surfactant protein A: a surface-plasmon-resonance study
Authors: SIDOBRE Stéphane , PUZO Germain , RIVIÈRE Michel
Published: July 2002

The human pulmonary surfactant protein A (hSP-A), a member of the mammalian collectin family, is thought to play a key defensive role against airborne invading pulmonary pathogens, among which is Mycobacterium tuberculosis, the aetiologic agent of tuberculosis. hSP-A has been shown to promote the uptake and the phagocytosis of pathogenic bacilli through the recognition and the binding of carbohydrate motifs on the invading pathogen surface. Recently we identified lipomannan and mannosylated lipoarabinomannan (ManLAM), two major mycobacterial cell-wall lipoglycans, as potential ligands for binding of hSP-A. We demonstrated that both the terminal mannose residues and the fatty acids are critical for binding, whereas the inner arabinosyl and mannosyl domains do not participate. In the present study we developed a surface-plasmon-resonance assay to analyse the molecular basis for the recognition of ManLAM by hSP-A and to try to define further the role of the lipidic aglycone moiety. Binding of ManLAM to immobilized hSP-A was consistent with the simplest one-to-one interaction model involving a single class of carbohydrate-binding site. This observation strongly suggests that the lipid moiety of ManLAM does not directly interact with hSP-A, but is rather responsible for the macromolecular organization of the lipoglycan, which may be necessary for efficient recognition of the terminal mannosyl epitopes. The indirect, structural role of the lipoglycan lipidic component is further supported by the complete lack of interaction with hSP-A in the presence of a low concentration of mild detergent.
Sheep have an unusual variant of the brain-specific metallothionein, metallothionein-III
Authors: CHUNG Roger S. , HOLLOWAY Adele F. , ECKHARDT Bedrich L. , HARRIS Julie A. , VICKERS James C. , CHUAH Meng Inn , WEST Adrian K.
Published: July 2002

Sheep metallothionein-III (MT-III) cDNA was isolated from a brain cDNA library and characterized. In contrast with MT-III from other species, sheep MT-III cDNA is predicted to encode a protein with significantly different metal-binding properties, owing to the loss of three of its cysteine residues. RT-PCR from other sheep confirmed that this aberrant structure is ubiquitous in this species. MT-III was successfully isolated from sheep brain, demonstrating that the cDNA does give rise to a protein product of the predicted structure. Sheep MT-III is similar to other mammalian MT-IIIs in that it retains the Cys-Pro-Cys-Pro motif which is thought to encode growth-inhibitory activity, and we show that it is likewise able to inhibit neuron survival in vitro. This is the first naturally occurring variant of MT-III (or any other major mammalian MT gene) which gives rise to a protein product. These findings are discussed in light of proposed roles of MT in the mammalian brain.
Regulation of phagosomal iron release from murine macrophages by nitric oxide
Authors: MULERO Victoriano , WEI Xiao-qing , LIEW Foo Y. , BROCK Jeremy H.
Published: July 2002

The role of NO in macrophage iron turnover was studied in macrophages from inducible nitric oxide synthase (iNOS)-deficient mice. Interferon γ/lipopolysaccharide (IFNγ/LPS)-activated bone marrow-derived macrophages from iNOS-deficient mice, following phagocytosis of 59Fe-labelled transferrin—anti-transferrin immune complexes, showed reduced iron release compared with cells from wild-type iNOS littermates. Uptake of the complexes by macrophages was similar in iNOS-deficient and wild-type mice. Ferritin was up-regulated by IFNγ/LPS treatment, but NO exercised a modest opposing down-regulatory effect. No effect of iNOS deficiency was seen when iron was taken up from iron citrate, which enters via a non-phagocytic route. These results suggest that NO plays a key role in regulating iron turnover in macrophages acquiring iron by phagocytosis of erythrocytes or cell debris, and thus the supply to peripheral tissues, such as to the bone marrow for erythropoiesis.
Characterization and expression of human bifunctional 3′-phosphoadenosine 5′-phosphosulphate synthase isoforms
Authors: FUDA Hirotoshi , SHIMIZU Chikara , LEE Young C. , AKITA Harukuni , STROTT Charles A.
Published: July 2002

Sulphonation, a fundamental process essential for normal growth and development, requires the sulphonate donor molecule 3′-phosphoadenosine 5′-phosphosulphate (PAPS), which is produced from ATP and inorganic sulphate by the bifunctional enzyme PAPS synthase. In humans, two genes encode isoenzymes that are 77% identical at the amino acid level, and alternative splicing creates two subtypes of PAPS synthase 2. The question as to whether distinctions in amino acid composition are reflected in differences in activity has been examined. The specific activity of the PAPS synthase 2 subtypes is 10- to 15-fold higher than that for PAPS synthase 1. The greater catalytic efficiency of the PAPS synthase 2 subtypes is demonstrated further by the 3- to 6-fold higher kcat/Km ratios for ATP and inorganic sulphate as compared with the ratios for PAPS synthase 1. In humans, PAPS synthase 1 is expressed ubiquitously, and is the dominant isoform in most tissues, whereas expression of the PAPS synthase 2 subtypes is variable and tissue-specific. It is noteworthy that, similar to other human tissues, PAPS synthase 1 also appears to be the dominant isoform expressed in cartilage. The latter finding initially created a conundrum, since there is a specific human dwarfing disorder that is known to be caused by a mutation in the PAPS synthase 2 gene. This apparent enigma would seem to be resolved by examination of cartilage from guinea-pigs as an animal model. Similar to humans, cartilage from mature animals predominantly expresses PAPS synthase 1. In contrast, expression of PAPS synthase 1 is relatively low in the cartilage of immature guinea-pigs, including the growth plate of long bones, whereas PAPS synthase 2 is the highly expressed isoenzyme.
Vps4-A (vacuolar protein sorting 4-A) is a binding partner for a novel Rho family GTPase, Rnd2
Authors: TANAKA Hiroko , FUJITA Hirotada , KATOH Hironori , MORI Kazutoshi , NEGISHI Manabu
Published: July 2002

Rho family GTPases are implicated in a variety of biological activities, including endocytic vesicle trafficking. Rnd2 is a new member of Rho family GTPases, but its biological functions are not known. In the present study, we have performed a yeast two-hybrid screening using Rnd2 as bait and revealed that Rnd2 binds specifically to Vps4-A (where Vsp4-A is vacuolar protein sorting 4-A), a member of the AAA ATPase family and a central regulator for early endosome trafficking. This interaction was determined by the yeast two-hybrid system, in vitro binding and co-immunoprecipitation studies. Vps4-A associated with both guanosine 5′-[β-thio]triphosphate-bound active and guanosine 5′-[β-thio]diphosphate-bound inactive forms of Rnd2. An ATPase-defective Vps4-A mutant, Vps4-AE228Q, expressed in HeLa cells was accumulated in the early endosomes. When Rnd2 was co-expressed with Vps4-AE228Q, Rnd2 was recruited to the Vps4-A-bound early endosomes. These results suggest that Rnd2 is involved in the regulation of endosomal trafficking via direct binding to Vps4-A.
The hyaluronan lyase of Streptococcus pyogenes bacteriophage H4489A
Authors: BAKER John R. , DONG Shengli , PRITCHARD David G.
Published: July 2002

Many pathogenic streptococci produce extracellular hyaluronan lyases which are thought to aid the spread of the organism in host tissues. In addition, several phages of group A streptococci are known to synthesize a bound form of hyaluronidase. It has been suggested that the function of this hyaluronidase is to facilitate penetration of the hyaluronan capsule by phage and thus to gain access for the phage to the cell surface of the host streptococcus [Hynes, Hancock and Ferretti (1995) Infect. Immun. 63, 3015–3020]. In the present work, the hyaluronidase of Streptococcus pyogenes bacteriophage H4489A, expressed in E. coli, has been purified and characterized. The enzyme was shown to be a lyase with a distributive action pathway. Unlike most bacterial hyaluronidases that have been characterized, the phage enzyme was found to specifically cleave hyaluronan, which adds credence to the view that its function is to digest the hyaluronan capsule of the host organism. This bacteriophage lyase may provide a practical alternative to the lyase from Streptomyces hyalurolyticus as a reagent for the specific cleavage of hyaluronan.
Proinsulin C-peptide activates cAMP response element-binding proteins through the p38 mitogen-activated protein kinase pathway in mouse lung capillary endothelial cells
Authors: KITAMURA Takanori , KIMURA Kazuhiro , JUNG Bae Dong , MAKONDO Kennedy , SAKANE Naoki , YOSHIDA Toshihide , SAITO Masayuki
Published: July 2002

Proinsulin C-peptide has been reported to have some biological activities and to be possibly involved in the development of diabetic microangiopathy. In the present study, we examined the effects of C-peptide on the mitogen-activated protein kinase pathway in LEII mouse lung capillary endothelial cells. Stimulation of the cells with C-peptide increased both p38 mitogen-activated protein kinase (p38MAPK) and extracellular signal-regulated kinase (ERK1/2) activities and activity-related site-specific phosphorylation of the respective kinases in a concentration-dependent manner, but failed to activate c-Jun N-terminal kinase. Stimulation of the cells with C-peptide also induced site-specific phosphorylation of cAMP response element (CRE)-binding protein (CREB)/activating transcription factor 1 (ATF1), and thereby binding of these transcription factors to CRE. Among three CREB kinases tested, phosphorylation of mitogen-activated protein kinase-activated protein kinase 2 (MAPKAP-K2) was induced after stimulation with C-peptide. The phosphorylation of CREB, ATF1 and MAPKAP-K2 were inhibited by SB203580, a p38MAPK inhibitor, but not by PD98059, an ERK kinase inhibitor. These results indicate that C-peptide activates p38MAPK followed by MAPKAP-K2 to enhance DNA—CREB/ATF1 interactions.
P2X7 receptors activate protein kinase D and p42/p44 mitogen-activated protein kinase (MAPK) downstream of protein kinase C
Authors: BRADFORD Michelle D. , SOLTOFF Stephen P.
Published: July 2002

Protein kinase D (PKD), also called protein kinase Cμ (PKCμ), is a serine/threonine kinase that has unique enzymic and structural properties distinct from members of the PKC family of proteins. In freshly isolated rat parotid acinar salivary cells, extracellular ATP rapidly increased the activity and phosphorylation of PKD. The stimulation by ATP required high concentrations, was mimicked by the P2X7 receptor ligand BzATP [2′- and 3′-O-(4-benzoylbenzoyl)ATP], and was blocked by Mg2+ and 4,4′-di-isothiocyano-2,2′-stilbene disulphonate (DIDS), suggesting that activation of PKD was mediated by P2X7 receptors, which are ligand-gated non-selective cation channels. Phorbol ester (PMA) and the activation of muscarinic and substance P receptors also increased PKD activity. PKC inhibitors blocked ligand-dependent PKD activation and phosphorylation, determined by in vitro phosphorylation studies and by phospho-specific antibodies to two activation loop sites (Ser744 and Ser748) and an autophosphorylation site (Ser916). ATP and BzATP also increased the tyrosine phosphorylation and activity of PKCΔ, and these stimuli also increased extracellular signal-regulated protein kinase (ERK) 1/2 activity in a PKC-dependent manner. PKD activation was not promoted by pervanadate (an inhibitor of tyrosine phosphatases) and was not blocked by PP1 (an inhibitor of Src family kinases) or genistein (a tyrosine kinase inhibitor), suggesting that tyrosine kinases and phosphatases did not play a major role in PKD activation. P2X7 receptor-mediated signalling events were not dependent on Ca2+ entry. These studies indicate that PKC is involved in cellular signalling initiated by P2X7 receptors as well as by G-protein-coupled receptors, and demonstrate that PKD and ERK1/2 are activated in similar PKC-dependent signalling pathways initiated by these diverse receptor types.
Phagosome maturation: aging gracefully
Authors: VIEIRA Otilia V. , BOTELHO Roberto J. , GRINSTEIN Sergio
Published: July 2002

Foreign particles and apoptotic bodies are eliminated from the body by phagocytic leucocytes. The initial stage of the elimination process is the internalization of the particles into a plasma membrane-derived vacuole known as the phagosome. Such nascent phagosomes, however, lack the ability to kill pathogens or to degrade the ingested targets. These properties are acquired during the course of phagosomal maturation, a complex sequence of reactions that result in drastic remodelling of the phagosomal membrane and contents. The determinants and consequences of the fusion and fission reactions that underlie phagosomal maturation are the topic of this review.