bluuebunny/crossref_metadata_2025 huggingface dataset:
Biochemical similarity of Schizosaccharomyces pombe ras1 protein with RAS2 protein of Saccharomyces cerevisiae
Authors: Onozawa Takashi , Danjoh Inaho , Fujiyama Asao
Published: May 2005

Schizosaccharomyces pombe contains single ras oncogene homologue, ras1, that functions in the signal transduction pathway conducting the cell's mating processes. To understand the biochemical basis of yeast ras proteins, we have purified the ras1 protein and compared the major biochemical constants with those of RAS2 protein from Saccharomyces cerevisiae and mammalian ras proteins. The purified ras1 protein showed a remarkably high Kd value for GDP binding (178 nM) and for binding with ATP. In contrast, the Kd value for GTP binding and the rate of GTPase activity were 64 nM and 77 × 10−6 s−1 at 37°C, respectively; both were higher than normal p21ras protein, but at the same level as the RAS2 protein. We directly measured rate of GTP binding and GDP binding which were 3.9 × 10−3 s−1 and 1.8 × 10−3 s−1 at 30°C, respectively. On the other hand, exchange rates between bound and free nucleotides remained almost constant throughout the tested combination of GTP and GDP, and were several‐fold lower than the binding rate. These results suggest that the release of the guanine nucleotide is the rate‐limiting step in the ras–GTP/GDP cycle. As a whole, the biochemical properties of the ras1 protein are close to those of the RAS2 protein, although these two proteins function differently in the signal transduction pathway in the cells.
The red ade mutants of Kluyveromyces lactis and their classification by complementation with cloned ADE1 or ADE2 genes from Saccharomyces cerevisiae
Authors: Zonneveld B. J. M. , van der Zanden A. L.
Published: May 2005

Seventy‐six red adenine mutants of Kluyveromyces lactis were isolated. By complementation they could be assigned to two groups with 31 and 45 mutants. Transformation of several strains from each group with plasmids containing the Saccharomyces cerevisiae ADE1 or ADE2 gene showed that the largest group was ade2 and the other group was ade1. Several previously isolated ‘ade1’ mutants were classified to either group and given new gene and allele numbers. ADE1 was localized at chromosome III, closely linked to the mating type gene, making it a convenient marker for mating type. ADE2 was localized at chromosome V.
Isolation and characterization of a novel yeast gene, ATH1, that is required for vacuolar acid trehalase activity
Authors: Destruelle Monika , Holzer Helmut , Klionsky Daniel J.
Published: May 2005

We have isolated a plasmid containing a gene, ATH1, that results in eight‐ to ten‐fold higher acid trehalase activity in yeast cells when present in high copy. The screening procedure was based on overproduction‐induced mislocalization of acid trehalase activity; overproduction of vacuolar enzymes that transit through the secretory pathway leads to secretion to the cell surface. A DNA fragment that confers cell surface expression of acid trehalase activity was cloned and sequenced. The deduced amino acid sequence displayed no homology to known proteins, indicating that we have identified a novel gene. A deletion in the genomic copy of the ATH1 gene eliminates vacuolar acid trehalase activity. These results suggest that ATH1 may be the structural gene encoding vacuolar acid trehalase or that the gene product may be an essential regulatory component involved in control of trehalase activity. The sequence has been deposited in the GenBank data library under Accession Number X84156 S. cerevisiae ATH1 gene.
Yeast sequencing reports. Gcs1, a gene encoding γ‐glutamylcysteine synthetase in the fission yeast Schizosaccharomyces pombe
Authors: Coblenz Anke , Wolf Klaus
Published: May 2005

By complementation of a mutant resistant to N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG) we have identified the gcs1 gene, encoding a putative γ‐glutamylcysteine synthetase. The gene is possibly interrupted by two introns and has 49% identical and 80% similar amino acids compared with the homologous protein from rat. In comparison with the Saccharomyces cerevisiae homologue it possesses 41% identical and 74% similar amino acids. The gsc1 sequence appears in the EMBL database under Accession Number X 85017.
Use of polymerase chain reaction epitope tagging for protein tagging in Saccharomyces cerevisiae
Authors: Schneider B. L. , Seufert W. , Steiner B. , Yang Q. H. , Futcher A. B.
Published: May 2005

Epitope tagging is the insertion of a short stretch of amino acids constituting an epitope into another protein. Tagged proteins can be identified by Western, immunoprecipitation and immunofluorescence assays using pre‐existing antibodies. We have designed vectors containing the URA3 gene flanked by direct repeats of epitope tags. We use the polymerase chain reaction (PCR) to amplify the tag‐URA3‐tag cassette such that the ends of the PCR fragments possess homology to the gene of interest. In vivo recombination is then used to direct integration of the fragment to the location of interest, and transformants are selected by their Ura+ phenotype. Finally, selection for Ura− cells on 5‐fluoro‐orotic acid plates yields cells where recombination between the repeated epitopes has ‘popped out’ the URA3 gene, leaving a single copy of the epitope at the desired location. PCR epitope tagging (PET) provides a rapid and direct technique for tagging that does not require any cloning steps. We have used PET to tag three Saccharomyces cerevisiae proteins, Cln1, Sic1 and Est1.
PMT3 and PMT4, two new members of the protein‐O‐mannosyltransferase gene family of Saccharomyces cerevisiae
Authors: Immervoll Thomas , Gentzsch Martina , Tanner Widmar
Published: May 2005

Two genes PMT3 and PMT4 were identified by polymerase chain reaction of genomic DNA using primers derived from regions of high homology between the products of three genes PMT1, PMT2 of Saccharomyces cerevisiae and part of a PMT1 related sequence of Kluyveromyces lactis. Pmt1p and Pmt2p are mannosyltransferases involved in the transfer of a mannosyl residue from dolichyl phosphate‐D‐mannose (Dol‐P‐Man) to seryl and threonyl residues in proteins. The products encoded by the PMT3 and PMT4 genes have almost identical hydropathy profiles in comparison to PMT1 and PMT2: a hydrophobic N‐ and C‐terminal third each with multiple potential transmembrane helices and a central hydrophillic part.

The predicted Pmt3p contains 753 amino acids, four potential N‐glycosylation sites and it is significantly homologous to Pmt1p, Pmt2p and Pmt4p. Pmt4p contains 762 amino acids and two potential N‐glycosylation sites. Northern blot analysis showed a single mRNA transcript of PMT3 and PMT4 of 2·8 kb. Thus PMT3 and PMT4 are two new members of the PMT gene family. The pmt4 null mutant the pmt3 pmt4 double null mutant, but not pmt3 null mutant, showed a small but significant shift of chitinase due to under glycosylation of the enzyme. The triple disruption pmt2 pmt3 pmt4 and the quadruple disruption result in a lethal phenotype. The EMBL data library Accession Number for PMT3 is X83797 and for PMT4 X83798.
An overview of membrane transport proteins in Saccharomyces cerevisiae
Authors: André Bruno
Published: May 2005

All eukaryotic cells contain a wide variety of proteins embedded in the plasma and internal membranes, which ensure transmembrane solute transport. It is now established that a large proportion of these transport proteins can be grouped into families apparently conserved throughout organisms. This article presents the data of an in silicio analysis aimed at establishing a preliminary classification of membrane transport proteins in Saccharomyces cerevisiae. This analysis was conducted at a time when about 65% of all yeast genes were available in public databases. In addition to ∼60 transport proteins whose function was at least partially known, ∼100 deduced protein sequences of unknown function display significant sequence similarity to membrane transport proteins characterized in yeast and/or other organisms. While some protein families have been well characterized by classical genetic experimental approaches, others have largely if not totally escaped characterization. The proteins revealed by this in silicio analysis also include a putative K+ channel, proteins similar to aquaporins of plant and animal origin, proteins similar to Na+‐solute symporters, a protein very similar to electroneural cation‐chloride co‐transporters, and a putative Na+‐H+ antiporter. A new research area is anticipated: the functional analysis of many transport proteins whose existence was revealed by genome sequencing.
Processing of pre‐ribosomal RNA in Saccharomyces cerevisiae
Authors: Venema Jaap , Tollervey David
Published: May 2005

Post‐transcriptional processing of precursor‐ribosomal RNA comprises a complex pathway of endonucleolytic cleavages, exonucleolytic digestion and covalent modifications. The general order of the various processing steps is well conserved in eukaryotic cells, but the underlying mechanisms are largely unknown. Recent analysis of pre‐rRNA processing, mainly in the yeast Saccharomyces cerevisiae, has significantly improved our understanding of this important cellular activity. Here we will review the data that have led to our current picture of yeast pre‐rRNA processing.
An Experimental Study of Ring Avulsion Injuries and Two Preventive Devices
Authors: DUBERT Th. , DIOP A. , VOELTZEL P.
Published: May 2005

We have performed biomechanical experiments on fresh cadaveric fingers to test two modifications to normal rings which may prevent ring avulsion injuries. One of these modifications produced reliable protection against ring avulsion accidents and was undetectable. It thus does not reduce the symbolic value of a wedding ring.
Degenerative Changes at the Scaphotrapezial Joint Following Herbert Screw Insertion: A Radiographic Study Comparing Patients with Scaphoid Fracture and Primary Hand Arthritis
Authors: NICHOLL J. E. , BUCKLAND-WRIGHT J. C.
Published: May 2005

One method of Herbert screw insertion is to mobilize the scaphotrapezial joint and insert the screw through the articular surface of the distal scaphoid. Because of concern that this might predispose to osteoarthritis in the scaphotrapezial joint, we have investigated joint space width and the presence of osteophytes adjacent to the scaphotrapezial joint using high definition macroradiography.

Twenty-eight patients with symptomatic primary osteoarthritis of their hands, 23 with a scaphoid fracture successfully treated conservatively and 18 treated with a Herbert screw were studied. The patients with primary osteoarthritis of their hands had more narrowing of the scaphotrapezial joint compared with the other two groups, but patients treated with a Herbert screw had a significantly higher incidence of osteophytosis on the distal scaphoid than the other two groups. With Herbert screw fixation, osteophytosis on the distal scaphoid may cause impingement and pain, and be a predictor of further degenerative joint changes.
The Treatment of Scaphoid Nonunion
Authors: MARUTHAINAR N. , RASQUINHA V. J. , GALLAGHER P.
Published: May 2005

We have retrospectively reviewed the results of 40 consecutive patients with nonunion of the scaphoid treated by the senior author (PG) between 1993 and 1996. These comprised two groups of patients. Group 1 comprised 20 patients treated between 1993 and 1994, with a Herbert screw and autograft, and Group 2, which also comprised 20 patients treated with the precision bone grafting technique which we describe. The precision bone grafting technique employs simple instrumentation to harvest bone percutaneously from the iliac crest and then insert it at the scaphoid nonunion site. The evaluation consisted of a clinical and radiological assessment of union and wrist function. Our review demonstrates a higher rate of union with the precision bone grafting technique than by the Herbert screw fixation with bone grafting.
The Value of the History in the Diagnosis of Carpal Tunnel Syndrome
Authors: BLAND J. D. P.
Published: May 2005

Details of the clinical history were elicited by questionnaire from 8,223 patients with suspected carpal tunnel syndrome and compared with the neurophysiological findings. Distribution of symptoms to the radial part of the hand and nocturnal exacerbation of symptoms showed the strongest individual correlations with positive nerve conduction studies. The regression model derived from the complete questionnaire achieved an overall sensitivity of 79% and specificity of 55% for the diagnosis of carpal tunnel syndrome when compared with the nerve conduction study results as a gold standard. A simple regression model for evaluating the history compares favourably with widely used clinical signs in its ability to predict the findings of nerve conduction studies.
Cubital Tunnel Syndrome: A Retrospective Review of 53 Anterior Subcutaneous Transpositions
Authors: LASCAR T. , LAULAN J.
Published: May 2005

Fifty-three cases of cubital tunnel syndrome were treated by anterior subcutaneous transposition of the ulnar nerve. All patients were assessed by an independent examiner at a mean follow-up of 32 months. McGowan’s rating scale, as modified by Goldberg, was used preoperatively and at follow-up. Preoperatively, five cases were classified grade I, 37 grade IIA, eight grade IIB and three grade III. Thoracic outlet syndrome was also present in 7 cases. At follow-up, 44 cases were grade 0, three grade I, five grade IIA and one grade IIB. Forty-four of the 53 cases had resolved and the other nine had improved. Subcutaneous transposition is a reliable and effective surgical option. The result is less satisfactory if a thoracic outlet syndrome is also present.
Does High-Resolution MR Imaging Have Better Accuracy than Standard MR Imaging for Evaluation of the Triangular Fibrocartilage Complex?
Authors: KATO H. , NAKAMURA R. , SHIONOYA K. , MAKINO N. , IMAEDA T.
Published: May 2005

We compared the findings of high-resolution MR imaging and standard MR imaging in the detection of tears of the triangular fibrocartilage in 33 patients with chronic wrist pain on the ulnar side. With arthroscopy as the standard of reference, sensitivity was 100%, specificity 53%, and accuracy 79% with the high-resolution MR imaging, against 83%, 67%, and 76% with the standard MR imaging. High-resolution MR imaging showed a higher sensitivity, but a decreased specificity in the assessment of the triangular fibrocartilage. The results showed that diagnosis of tears in the triangular fibrocartilage by MR imaging, even high-resolution MR imaging, is unsatisfactory, although further technological advances may well improve the accuracy.
A Stress Distribution Analysis of a Ceramic Lunate Replacement for Kienböck’s Disease
Authors: ODA M. , HASHIZUME H. , MIYAKE T. , INOUE H. , NAGAYAMA N.
Published: May 2005

To determine the usefulness of a ceramic lunate replacement for the wrist joint as a possible treatment for Kienböck’s disease, stress distribution of five wrist joint models (a normal wrist, one with the lunate bone removed, one with a coiled tendon replacement, one with a coiled tendon replacement with a ceramic core, and one with a ceramic lunate replacement coated with sheets of tendon) were analysed by the finite element method. The results of finite element method analyses were compared with those of pressure distribution analyses on the wrist joint using pressure-sensitive film and measurement of pressure on the radio-lunate joint with a tactile sensor. Both the coiled tendon model and ceramic core model were unable to resist applied force. Stress and pressure were well-distributed in the wrist joint of the ceramic lunate implant model. These results indicate that the ceramic lunate implant could be expected to prevent carpal collapse.
Diagnostic Ultrasonography of the Ulnar Nerve in Cubital Tunnel Syndrome
Authors: OKAMOTO M. , ABE M. , SHIRAI H. , UEDA N.
Published: May 2005

Thirty-two elbows in 31 patients diagnosed as having cubital tunnel syndrome underwent ultrasonographic examination to assess morphological changes in the ulnar nerve and its surrounding tissues. On longitudinal images, the site of constriction due to the fibrous band and proximal swelling of the nerve were observed by ultrasonography and were confirmed intraoperatively. On axial images, the lengths of the major axis [7.2 (SD 1.6) mm] and the minor axis [3.7 (0.9) mm] of the nerve at the medial epicondyle were greater than those in normal subjects. There was a correlation between the stage of ulnar nerve palsy and the diameter of the major axis. Preoperatively, ganglia were detected by ultrasonography in the cubital tunnel in three cases and an anconeus epitrochlearis muscle in two.
Ultrasonography in the Diagnosis of Ulnar Tunnel Syndrome Caused by an Occult Ganglion
Authors: NAKAMICHI K. , TACHIBANA S. , KITAJIMA I.
Published: May 2005

We report a case of ulnar tunnel syndrome caused by an occult ganglion which was diagnosed preoperatively by ultrasonography.
The Diagnostic Value of Intraoperative Measurement of Choline Acetyltransferase Activity During Brachial Plexus Surgery
Authors: HATTORI Y. , DOI K. , FUKUSHIMA S. , KANEKO K.
Published: May 2005

We used the measurement of choline acetyltransferase (CAT) activity to distinguish between preganglionic and postganglionic injuries of the spinal roots during brachial plexus surgery. This study includes ten spinal roots in six patients with traumatic brachial plexus injuries. The results of CAT activity were compared with myelographic findings, the operative findings and the intraoperative evoked spinal cord potentials. The results of CAT activity and the operative findings were consistent in all roots. Whereas CAT activity in fascicles in preganglionic injured roots was below 500 cpm, it was above 2000 cpm in postganglionic injured roots. This study shows that intraoperative measurement of CAT activity can provide useful information on the site and severity of brachial plexus injuries and the assessment of the motor function status of the injured spinal root, both directly and quantitatively.
Are Mestizos Hybrids? The Conceptual Politics of Andean Identities
Authors: DE LA CADENA MARISOL
Published: May 2005

Through a genealogical analysis of the terms mestizo and mestizaje, this article reveals that these voices are doubly hybrid. On the one hand they house an empirical hybridity, built upon eighteenth and nineteenth century racial taxonomies and according to which ‘mestizos’ are non-indigenous individuals, the result of biological or cultural mixtures. Yet, mestizos’ genealogy starts earlier, when ‘mixture’ denoted transgression of the rule of faith, and its statutes of purity. Within this taxonomic regime mestizos could be, at the same time, indigenous. Apparently dominant, racial theories sustained by scientific knowledge mixed with, (rather than cancel) previous faith based racial taxonomies. ‘Mestizo’ thus houses a conceptual hybridity – the mixture of two classificatory regimes – which reveals subordinate alternatives for mestizo subject positions, including forms of indigeneity.
History, Sociology and the Political Conflicts of the 1920s in São Paulo, Brazil
Authors: WOODARD JAMES P.
Published: May 2005

In recent years an ‘ethno-economic’ interpretation of politics in the coffee state of São Paulo in the early twentieth century, associated particularly with the work of Mauricio Font, has gained widespread acceptance. Its claim that state politics in the period was increasingly shaped by a cleavage between a declining traditional coffee aristocracy on the one hand, and a rising, mostly immigrant, smallholding and industrial economy on the other, is challenged here. It is argued, in part on the basis of a re-examination of the sources used by Font, that ideological rather than economic concerns motivated the Liga Nacional, while at county level, in Araras and elsewhere, personal and clientelistic motives continued to shape political loyalties. Finally, the argument that the Partido Democrático was driven primarily by ‘Big Coffee’ reaction to the PRP is shown to be unfounded.
Round spermatids stained with MitoTracker can be used to produce offspring more simply
Authors: Hikichi Takafusa , Kishigami Satoshi , Van Thuan Nguyen , Ohta Hiroshi , Mizutani Eiji , Wakayama Sayaka , Wakayama Teruhiko
Published: May 2005

Although both intracytoplasmic sperm injection (ICSI) and round spermatid injection (ROSI) are used in infertility treatments, the rate of offspring achieved with ROSI is low compared with that achieved with ICSI. The difficulty in correctly selecting round spermatids from testicular cells is one of the causes of this phenomenon. We easily selected live round spermatids from testicular cells stained with 20 nM MitoTracker, which visualizes mitochondria without killing the cell. Using this method, we divided round spermatids into three groups based on the polarization of their mitochondria, and performed ROSI. The rate of successful offspring achieved with MitoTracker-stained ROSI was the same in all groups. This indicates that changes in the polarization of mitochondria in round spermatids are not directly related to the developmental capacity of subsequently fertilized embryos. Because this staining has no harmful effects on embryo development, the selection of spermatids by MitoTracker under a fluorescence microscope should be useful in research into and the treatment of infertility.
Influence of seminal plasma PSP-I/PSP-II spermadhesin on pig gamete interaction
Authors: Caballero Ignacio , Vázquez Juan M. , Rodríguez-Martínez Heriberto , Gil Maria A. , Calvete Juan J. , Sanz Libia , Garcia Eva M. , Roca Jordi , Martínez Emilio A.
Published: May 2005

The seminal plasma PSP-I/PSP-II spermadhesin is able to preserve, in vitro, the viability of highly extended boar spermatozoa, suggesting it might be used as a suitable ameliorator for the damaging effects of sperm handling, including in vitro fertilization. However, little is known about the ligand capability of PSP-I/PSP-II as regards the zona pellucida (ZP) or its possible role in gamete interaction. The present study evaluated the effect of the presence of PSP-I/PSP-II (1.5 mg/ml) during in vitro oocyte maturation and also during co-incubation of frozen-thawed boar spermatozoa with either immature (IM) or in vitro matured (IVM) oocytes, either enclosed by cumulus cells or denuded. Exposure of the gametes to the heterodimer during in vitro gamete co-incubation showed a significant blocking effect of sperm penetration rates and a decreased number of spermatozoa per oocyte in both IM and IVM denuded oocytes. Such an effect was not present in cumulus-enclosed oocytes, suggesting the effect could be mediated by exposed ZP receptors. In addition, when PSP-I/PSP-II was added to the IVM medium, oocyte maturation rates were significantly reduced. In conclusion, the results suggest that PSP-I/PSP-II, when present in vitro, blocks sperm–ZP binding.
Production of rabbit chimeric embryos by aggregation of zona-free nuclear transfer blastomeres
Authors: Chrenek P. , Makarevich A.V.
Published: May 2005

The objective of this study was to compare in vitro developmental capacity of zona-free aggregated rabbit chimeric embryos and the allocation of EGFP (enhanced green fluorescence protein) gene expression to the inner cell mass (ICM). We produced chimeric embryos by synchronous aggregation of zona-free blastomeres from embryonic cell nuclear transfer (EMB-NT) or somatic cell nuclear transfer (SC-NT) and blastomeres from normal zona-free embryos (N) at the 16-cell stage. In the control group, transgenic (TR) and normal zona-free embryos were used to produce chimeric embryos (TR<>N). EMB-NT embryos were produced by fusion of enucleated oocytes with embryonic cells, which were derived from 32-cell stage transgenic embryos bearing the EGFP gene. The SC-NT embryos were produced by fusing enucleated oocytes with cumulus cells, which were derived from homozygotes transgenic for the EGFP gene female oocytes at 16 h post-coitum. Nuclei of transgenic blastomeres emitted a green signal under fluorescence microscopy. Zona-free EMB-NT or zona-free SC-NT rabbit embryos, both with EGFP fluorescence, as well as TR and zona-free rabbit embryos with no fluorescence (EMB-NT<>N, SC-NT<>N, TR<>N) were aggregated on day 2.5 and evaluated on day 5. The proportion of EMB-NT<>N embryos that developed to the blastocyst stage was significantly higher compared with SC-NT derived cells (p<0.05), but significantly lower than in TR<>N chimeric blastocysts (p<0.001). Similarly, a higher proportion (p<0.001) of EGFP-positive cells allocated to ICM of chimeric blastocysts was revealed in TR<>N chimeras (55%), compared with EMB-NT<>N (35%) and SC-NT<>N (21%). Our results indicate that synchronous chimeric embryos reconstructed from TR embryos were better able to develop and colonize the ICM area than EMB-NT and SC-NT embryos. In this study we have demonstrated for the first time that rabbit NT-derived embryos are able to develop into chimeric blastocysts and participate in the ICM area.
The Penn Chinese TreeBank: Phrase structure annotation of a large corpus
Authors: XUE NAIWEN , XIA FEI , CHIOU FU-DONG , PALMER MARTA
Published: May 2005

With growing interest in Chinese Language Processing, numerous NLP tools (e.g., word segmenters, part-of-speech taggers, and parsers) for Chinese have been developed all over the world. However, since no large-scale bracketed corpora are available to the public, these tools are trained on corpora with different segmentation criteria, part-of-speech tagsets and bracketing guidelines, and therefore, comparisons are difficult. As a first step towards addressing this issue, we have been preparing a large bracketed corpus since late 1998. The first two installments of the corpus, 250 thousand words of data, fully segmented, POS-tagged and syntactically bracketed, have been released to the public via LDC (www.ldc.upenn.edu). In this paper, we discuss several Chinese linguistic issues and their implications for our treebanking efforts and how we address these issues when developing our annotation guidelines. We also describe our engineering strategies to improve speed while ensuring annotation quality.
The Union and Turkey: The Hard Way
Authors:   JWS/WTE
Published: May 2005

On November 3, 2002, the Turkish elections brought an overwhelming victory for the Justice and Development Party (Adalet ve Kalkınma Partisi-AKP), the first party since 1987 to secure a clear majority in Parliament. It triumphed on a ticket including Turkey's entry into the European Union. Valéry Giscard d'Estaing then grasped the occasion and his newfound status of Convention chairman to vent an old conviction (reported in Le Monde of 9 November) that Turkey's entry into the EU would be ‘the end of European Union’. It was a good thing that hardly anyone in the Convention took notice of this irrelevant and even irresponsible step. But in the eyes of the public a link between the Constitution and Turkey's accession had been established beyond doubt. It could only be undone by European leaders in power (first by Chirac) at the cost of heavy concessions to populist demands. France will have a separate referendum on Turkey and on other accessions thanks to a constitutional amendment (see Carcassonne's contribution to this issue). Other countries will follow no doubt.
The Barroso Drama: Reality for the EU Constitution
Authors:   WTE
Published: May 2005

In importance for the Union's constitutional evolution, the October 2004 events at least equal the European Constitutional Convention, including the IGC. This is the idea behind the following tens of pages in this issue of EuConst devoted to their analysis. Eight authors have been involved in putting together this story, seven of whom are young researchers from different countries in the Union. Their participation allowed many national aspects of the drama to be brought out in a way which otherwise would have been impossible.
The Barroso Drama: France: The Jacques Barrot Way
Authors: Mariatte Flavien
Published: May 2005

Fluctuating French media attention for Barroso's drama. Chirac's support for Manuel Barroso. Appointment of European Commission and French Constitution. French debate on candidacy Barrot and the assigned transport portfolio. Offer extreme right party to save Barroso's team. The unmentioned embezzlement.
National Constitutional Concepts in the New Constitution for Europe
Authors: Ziller Jacques
Published: May 2005

Notion of constitutional concept varies over time and space. Constitution for Europe as a further step in guaranteeing rights and separating powers. Amongst others French, Italian, German and Swedish national concepts in the Treaties of the European Communities and of the European Union. Several causes for difficult tracing of national concepts in the Constitution for Europe. Convention method: instead of by diplomats and EU experts, drafting by members of Parliament and (former) members of Government. More room for national concepts. The Intergovernmental Conference: the Empire strikes back, but with mixed impact. Parallels with other constitution making procedures. Concept of constitution: structure and size no argument for denying constitutional character; Constitution octroyée v. contrat social; Franco-American revolution v. British tradition.
Constitutional Debates on Parliamentary Inviolability in Turkey
Authors: Özbudun Ergun
Published: May 2005

Differences between parliamentary inviolability and parliamentary non-liability. Parliamentary inviolability in the Turkish constitution. Also for ministers. Scope and duration. Lifting of inviolability by the Grand National Assembly. Possibility of judicial review. Constitutional amendments to limit inviolability in the combat against corruption rejected.
France Conseil Constitutionnel on the Status of (Secondary) Community Law in the French Internal Order. Decision of 10 June 2004, 2004-496 DC
Authors: Reestman Jan Herman
Published: May 2005

Shortly before the European Council reached political agreement on the European Constitutional Treaty (18 June 2004), the Conseil constitutionnel [the French constitutional court], rendered a decision on the relationship between (secondary) community law and the French Constitution that will still be pivotal once the Constitutional Treaty has entered into force. Taken on 10 June 2004, the decision was only made public five days later. The Conseil feared that it would be presented as ‘a French abdication in the face of the European institutions’ and might influence the elections for the European Parliament of 13 June 2004.
Energy landscape of amyloidogenic peptide oligomerization by parallel-tempering molecular dynamics simulation: Significant role of Asn ladder
Authors: Tsai Hui-Hsu (Gavin) , Reches Meital , Tsai Chung-Jung , Gunasekaran Kannan , Gazit Ehud , Nussinov Ruth
Published: May 2005

Recent evidence suggests that amyloidogenic oligomers may be the toxic species in cell cultures. Thus, it is crucial to understand their structure and oligomerization mechanism in atomistic detail. By employing tens of fast central processing units and an advanced phase-space sampling algorithm, parallel-tempering molecular dynamics, we have explored the energy landscape of amyloidogenic peptide oligomerization in explicit water. A pentapeptide, DFNKF, derived from human calcitonin and its mutant, DF
A
KF, was simulated with a total simulation time of ≈500 ns. The detailed oligomerization process of a DFNKF parallel β-sheet formation at 300 K has been characterized. The assembly of a parallel β-sheet from the amorphous state mainly occurs via a “bottleneck” channel where the interstrand Asn–Asn stacking is the major interaction. The interactions of Asn–Asn stacking include both backbone and side-chain hydrogen bonds. The Asn–Asn interactions work like “glue” by sticking the DFNKF strands together and assist the “on-pathway” oligomerization. The Asn–Asn stacking observed here is similar to the
Asn
ladder commonly found in globular β-helix proteins. A control run shows that when Asn is mutated to Ala, the stability and population of the DF
A
KF parallel β-sheet is decreased. Furthermore, our
in vitro
mutagenesis experiments show that the ability of DF
A
KF peptides to form amyloid fibrils is significantly reduced, in agreement with the simulations. Knowledge of the energy landscape of oligomerization may provide hints for rational drug design, preventing amyloid-associated diseases.
Cell cycle inhibition provides neuroprotection and reduces glial proliferation and scar formation after traumatic brain injury
Authors: Di Giovanni Simone , Movsesyan Vilen , Ahmed Farid , Cernak Ibolja , Schinelli Sergio , Stoica Bogdan , Faden Alan I.
Published: May 2005

Traumatic brain injury (TBI) causes neuronal apoptosis, inflammation, and reactive astrogliosis, which contribute to secondary tissue loss, impaired regeneration, and associated functional disabilities. Here, we show that up-regulation of cell cycle components is associated with caspase-mediated neuronal apoptosis and glial proliferation after TBI in rats. In primary neuronal and astrocyte cultures, cell cycle inhibition (including the cyclin-dependent kinase inhibitors flavopiridol, roscovitine, and olomoucine) reduced up-regulation of cell cycle proteins, limited neuronal cell death after etoposide-induced DNA damage, and attenuated astrocyte proliferation. After TBI in rats, flavopiridol reduced cyclin D1 expression in neurons and glia in ipsilateral cortex and hippocampus. Treatment also decreased neuronal cell death and lesion volume, reduced astroglial scar formation and microglial activation, and improved motor and cognitive recovery. The ability of cell cycle inhibition to decrease both neuronal cell death and reactive gliosis after experimental TBI suggests that this treatment approach may be useful clinically.
Nutrition and lysosomal activity. The effect of dietary cod-liver oil on the distribution of polyunsaturated fatty acids in the kidney lysosomes of rats receiving deficient or adequate intakes of vitamin E
Authors: Iqbal M. , Dingle J. T. , Moore T. , Sharman I. M.
Published: May 2005

1. In an attempt to explain the antagonism between polyunsaturated fatty acids (PUFA) and vitamin E in the promotion of post-mortem autolysis and increased lysosomal fragility in the kidney of rats, studies were made by gas chromatography of the incorporation of PUFA into the lysosomes.

2. Kidneys were taken from rats that had received various diets, which differed in their fat components and which were with or without vitamin E. Since the inclusion of cod-liver oil in the diet reduces the period of dietary preparation necessary for rapid kidney autolysis, the effect of this oil on the PUFA distribution in the lysosomes was specially studied.

3. In purified preparations of kidney lysosomes from rats that had received substantial amounts of cod-liver oil for several weeks, C 20:5 acid was incorporated mainly at the expense of C 18:2 (linoleic) and C 20:4 (arachidonic) acids. In less purified lysosomal fractions the incorporation of C 20:5 and C 22:6 acids and the corresponding reductions in linoleic and arachidonic were well advanced after 10 days, but were not maximal until about 30 days. The same changes took place in the reverse direction, with about the same rapidity, when rats that had previously been given cod-liver oil were changed to a diet containing lard.

4. The percentage of PUFA in the kidney lysosomes of rats not dosed with vitamin E was not significantly different from that of rats given adequate doses.

5. Thus the increased tendency to kidney autolysis, and the reduced stability of the lysosomes, caused by the feeding of cod-liver oil were associated with the partial replacement of linoleic acid and of the endogenous arachidonic acid by an acid, usually foreign to the rat, which is even more unsaturated. Since vitamin E did not prevent the entry of this acid into the lysosomes its potency in retarding autolysis and stabilizing the lysosomes must be exerted at some point subsequent to the incorporation of PUFA.

6. Since change in the percentage of lysosomal PUFA in response to dietary changes is slow, this suggests that the lysosornal lipidsescape usage in general metabolism as an immediate source of calories and that they have a half-life of at least 15 days. Individual fatty acids, however, may differ in their half-lives.
The fermentation of sugar-beet pulp and sucrose in an artificial rumen, and the effect of linseed oil fatty acids on the fermentation
Authors: Czerkawski J. W. , Breckenridge Grace
Published: May 2005

1. An artificial rumen is described together with associated analytical and sampling procedures.

2. The equipment was used to study the effects of linseed oil fatty acids on the fermentation of sugar-beet pulp and of sucrose by rumen micro-organisms.

3. With sugar-beet pulp the addition of small amounts of linseed oil fatty acids had little effect on total gas production, utilization of sugar and the molar proportions of steam-volatile acids formed, but they strongly inhibited methane production, whereas large amounts of the fatty acids temporarily inhibited the production of gas and utilization of sugar.

4. With sucrose even the smaller amounts of fatty acids had a marked inhibitory effect on the production of gas and utilization of sugar.

5. The technique is assessed critically and the results are discussed in the light of previous experiments in vivo.
Studies on magnesium in ruminant nutrition
Authors: Suttle N. F. , Field A. C.
Published: May 2005

1. In Expt 1, five groups of four dry non-pregant ewes were given 1 kg/day of a semi-purified diet containing either 0.05, 0.10 or 0.15% magnesium with 0 or 4.44% added potassium. Plasma Mg Concentrations were reduced by increasing the K intake (P < 0.001) and by reducing the Mg intake (P < 0.1). The effects were additive and, at the lowest Mg intake, supplementary K induced two cases of hypomagnesaemic tetany.

2. Expt 2 was similar to Expt 1 except that the ewes were suckling single lambs, they were given 2 kg food/day and the lowest dietary Mg concentration was 0.075%. K again lowered plasma Mg values and induced two cases of tetany but reductions in Mg intake had no effect.

3. After repletion on the basal diet, sixteen lactating ewes from Expt 2 were transferred to a diet containing 0.05% Mg with 0, 2.22 or 4.44% added K. Plasma Mg concentrations fell rapidly on each treatment; the effect was greatest in K-supplemented groups ( P < 0.05) and two cases of hypomagnesaemic tetany were induced at the highest K intake.

4. In each experiment the K supplements caused a rapid increase in plasma K concentrations to a relatively constant level some 5 mg/100 ml above the resting value of 18.0 mg/100 ml.

5. Some sheep refused K-supplemented diets at first and there was a tendency for refusals to be greatest at the lowest Mg intakes.

6. The results are discussed in relation to the aetiology of hypomagnesaemic tetany.
Increased retention of orally administered zinc and raised blood cell zinc concentrations in iron-deficient rats
Authors: Spry C. J. F. , Piper K. G.
Published: May 2005

1. Rats were made severely iron-deficient by feeding with Fe-deficient diets, which contained adequate amounts of zinc.

2. Ten days after an oral dose of 250 μg65Zn the Fe-deficient rats had retained 17% whereas control rats had retained only 8% of the dose.

3. When unlabelled Zn was given intravenously to the Fe-deficient and control rats there was a reduction in retention of the orally administered 65Zn. This effect was not produced when Fe was given before the oral dose of 65Zn.

4. Total body Zn, whole blood Zn and plasma Zn concentrations of the rats were measured by atomic absorption spectrophotometry. Both the Fe-deficient rats and the control rats had the same total body Zn calculated as a proportion of body-weight. The plasma Zn concentrations were normal in all the animals. Blood cell Zn concentrations in the Fe-deficient animals were raised.

5. The blood cell Zn concentration was directly related to the reticulocyte count in all the rats.

6. It was concluded that Fe and Zn are absorbed by different metabolic pathways, and that there is an increased turnover of Zn in the Fe-deficient rats, possibly related to the short lifespan and increased Zn concentration of Fe-deficient rat erythrocytes. In the rat abnormalities of Zn concentration can only be considered significant if Fe status is also known.
Effect of acetoacetate on urinary excretion of xanthurenic acid and other tryptophan metabolites in rats
Authors: Nath M. C. , Shastri N. V.
Published: May 2005

1. An experiment was performed to study the effect of sodium acetoacetate on urinary excretion of santhurenic acid and other tryptophan metabolites in male albino rats.

2. Animals were fed on a nicotinic acid-deficient diet for a period of 3 weeks. The animals were then divided into two groups and, after the basal urinary excretion of the tryptophan metabolites had been estimated, the rats of both the groups were force-fed with L-tryptophan (100 mg per rat), the rats of the second group being simultaneously injected intraperitoncally with acetoacetate (200 mg/kg body-weight), and the urine samples during the following 24 h were collected and analysed.

3. Acetoacetate-treated rats given tryptophan were found to excrete significantly greater amounts of kynurenine, hydroxykynurenine and xanthurenic acid than the corresponding control rats. There was no ditference between the amounts of kynurenic acid excreted by the animals in the two groups.
The effect of nutrition and androgens on the composition of bovine blood plasma and seminal plasma at puberty
Authors: Baronos S. , Mann T. , Rowson L. E. A. , Skinner J. D.
Published: May 2005

1. Experiments on three pairs of male identical twin calves have shown that underfeeding at 2.5–6 months of age, i.e. during the critical stage of sexual maturation, considerably delayed the process of puberty, as regards testicular androgen production as well as secretion of fructose and citric acid by the seminal vesicles.

2. There was no marked difference in the blood-plasma content of non-protein nitrogen, vitamin A and carotene at 6 months, as between twins on a normal (N) and low (L) plane of nutrition, although the L-twins were growing and maturing more slowly. However, the blood-plasma content of glucose and of most of the free amino acids, was slightly reduced as a result of underfeeding.

3. When at the age of 6 months two twin-pairs were castrated and injected with either testosterone or androstenedione (50 mg every 3rd day, for 2 months), the L-twins of both pairs and the N-twin of one pair grew at a faster rate. Moreover, the concentration of nitrogen in blood plasma increased, the rise being particularly evident in the content of indispensable amino acids. This effect, however, was more pronounced in the N- than in the L-twins.

4. Testosterone treatment enhanced libido in the N-twin but not in the corresponding L-twin. Androstenedione had no effect on libido in either the N-twin or the L-twin. Thus, although testosterone and androstenedione both effectively promoted growth and nitrogen metabolism, androstenedione, unlike testosterone, was devoid of androgenic activity.
Vitamin E and hepatotoxic agents
Authors: Green J. , Bunyan J. , Cawthorne M. A. , Diplock A. T.
Published: May 2005

1. It has been suggested that carbon tetrachloride damages rat liver by accelerating processes of lipid peroxidation at subcellular sites and that the protective action of vitamin E is due to its functioning as an antioxidant in vivo. Direct evidence for these mechanisms in vivo has been sought and is critically examined.

2. The increased production of malondialdehyde by rat liver microsomal fractions during incubation with CCl4 was shown to be a function of the vitamin E status of the rat and of an in vitro reaction, which could not be correlated with the hepatotoxic action of CCI4.

3. Evidence for the production of lipid peroxides by CCl4 in the livers of vitamin E-deficient and vitamin E-supplemented rats was sought (I) by measurement of ultraviolet spectral changes ('diene' formation) and (2) by direct micro-iodimetric determination of the peroxide. No differences in peroxide content were found between CC14-treated and control rats, nor were the spectrophotometric changes in the ultraviolet region related to the presence of vitamin E.

4. The effect of CCI4 (2.0 ml/kg orally) on ATP levels in rat liver was studied at intervals from 3 to 68 h. The primary lesion leading to necrosis and fat accumulation after CCl4 treatment occurred many hours before the eventual slight decline in ATP. Although the levels of ATP were somewhat higher in vitamin E-deficient rats, vitamin E did not prevent the slight decline in ATP that took place. Since ATP is known to be highly sensitive to peroxidation, the results suggest that lipid peroxidation is not the primary event in CCl4 poisoning.

5. The effect of CC14on the metabolism of [14C]D-α-tocopherol in the rat was studied. A single intraperitoneal dose of CCl4 (2.0 m/kg) did not increase the destruction of α-tocopherol in the liver or carcass after 24 h. Three smaller daily doses of CC14 (0.25 ml/kg) also did not increase α-tocopherol catabolism; on the contrary, significantly more α-tocopherol was found in the livers of rats treated with CCI4. These results suggest that CCl4 does not increase lipid peroxidation in vivo.
Studies on intestinal digestion in the sheep
Authors: Macrae J. C. , Armstrong D. G.
Published: May 2005

1. In seven experiments sheep were given diets ranging from all-hay to all-barley, and also a diet comprising one part hay to two parts flaked maize. Each sheep was equipped with a cannula into the rumen and a re-entrant cannula in the proximal duodenum; six of the ten also had a re-entrant cannula in the terminal ileum. Paper impregnated with chromic oxide (Cr2O3) was given twice daily by rumen fistula.

2. Amounts of α-linked glucose polymer entering and leaving the small intestine and excreted in the faeces were measured. Some values for a fraction designated non-glucose reducing polymer for cellulose and for gross energy were also obtained. The amounts passing the proximal duodenum and the terminal ileum were adjusted to give 100% recovery of Cr2O3and the values were used to measure the extent of digestion in various parts of the alimentary tract.

3. When rolled or whole barley was given alone or was the major feed constituent the amount of α-linked glucose polymer entering the small intestine was 6.0±0.76% of that ingested (range 2.6–8.1%). The value was significantly lower than that found for the diet of hay and flaked maize (10.4±1.3%, range 8.0–13.6 %).The α-linked glucose polymer which entered the small intestine was almost completely digested there.

4. The digestibility of the non-glucose reducing polymer, which included much of the hemicelluloses present, ranged from 51 to 73% and almost all the digestible fraction (93–97%) was digested before the small intestine when hay or predominantly hay diets were given. On high-cereal diets only 71–85% of the digested fraction disappeared before the small intestine and appreciable amounts were digested in the large intestine.

5. On the all-hay diet 91% of the digestible cellulose and 67% of the digestible energy were lost before the small intestine, 0 and 21% in the small intestine and 9 and 12% in the large intestine.

6. Mean digestibility coefficients determined in sheep fed solely on either whole or rolled barley were: for dry matter 88.1 and 87.9%, for nitrogen 83.5 and 82.1%, for crude fibre 53.7 and 56.6% and for gross energy 87.7 and 88.0%.