bluuebunny/crossref_metadata_2025 huggingface dataset:
The Arrest of Abdullah Öcalan
Authors: Gilbert Geoff
Published: March 2004

Abdullah Öcalan's arrival in Turkey in February 1999 followed a prolonged search in Europe for asylum following his expulsion from Syria in late 1998. His coming within Turkish jurisdiction raises questions about the international processes to bring alleged transnational fugitive offenders before the courts. This article looks at the extradition regime within Europe and the alternative methods of rendition that were eventually employed to remove him from Kenya. Extradition law developed during the nineteenth century and is based on ideas of revolution, the principle of nationality and liberal democracy which pervaded that period. The late twentieth century has a different ethos that offers fewer protection to the political revolutionary, but has incorporated international human rights standards. Extradition law straddles the enforcement of criminal law, non-interference in the domestic affairs of another State and international human rights law. The article concludes by examining the demands of international human rights law for the trial in Turkey.
The Brčko Final Award of 5 March 1999
Authors: Schreuer Christoph
Published: March 2004

The Dayton Accords of 1995 for Peace in Bosnia and Herzegovina provided for arbitration over the boundary line in the Brčko area of Bosnia. Arbitration took place between the Muslim and Croat Federation and the Republika Srpska. The Award of 14 February 1997 did not draw a boundary line but established an international interim supervisory regime. On 15 March 1998 the Arbitral Tribunal handed down a Supplemental Award which extended the interim regime pending further action by the Tribunal. The Final Award of 5 March 1999 establishes a permanent self-governing Brčko District government that is independent of the two entities. The Tribunal reserves the authority to revise its decision in case of a failure of either party to cooperate in the implementation of the new regime.
List of Current Proceedings: Update
Authors: Amaya-Castro Juan M.
Published: March 2004

Libya filed two separate applications on 3 March 1992. In the applications, Libya contended that it had not been possible to settle this dispute by negotiations and that the parties unable to agree on the organization of an arbitration to hear the matter. It accordingly submitted the disputes to the Court on the basis of Article 14(1) of the Montreal Convention. Libya refers in the applications to the charging of two Libyan nationals, by the Lord Advocate of Scotland, and by a Grand Jury of the United States, respectively, with having caused a bomb to be placed aboard a Pan-am flight, which bomb subsequently exploded, causing the aeroplane to crash. Libya contends that the United Kingdom and the United States, respectively, by rejecting the Libyan efforts to resolve the matter within the framework of international law, including the Montreal Convention, are pressuring it into surrendering the two Libyan nationals for trial. In this connection, Libya refers to Article 1 of the Montreal Convention, according to which the charge constitutes an offence, and to the several other articles of that Convention which are relevant to Libya's alleged right to jurisdiction over the matter and the prosecution thereof. Libya alleges that these obligations are breached by the United Kingdom and the United States respectively.
The Nuclear Weapons Advisory Opinion of the International Court of Justice and the Fundamental Distinction Between the Jus ad Bellum and the Jus in Bello
Authors: Gill Terry
Published: March 2004

The 1996 Nuclear Weapons Advisory Opinion of the International Court has been both hailed and criticized on various grounds. However, one area, namely the Court's treatment of the distinction between the law regulating the use of force and the humanitarian law of armed conflict, has received relatively little attention. This author is convinced and concerned that the Court's treatment of this issue misconstrued the relationship between these two branches of the law, and in doing so potentially weakened any restraining influence the law of armed conflict might have on the potential use of nuclear weapons.
Constitutional and Institutional Developments
Authors: Rydberg Åsa
Published: March 2004

By decision of the Judges at the Twentieth plenary session of the International Criminal Tribunal for the former Yugoslavia (ICTY) two new rules were adopted, Rule 77 bis and Rule 84 bis of the Rules of Procedure and Evidence (Rules). In addition, 13 rules were amended.
Commentary: Rule 61 Procedure in the International Criminal Tribunal for the Former Yugoslavia: A Lesson for the ICC
Authors: Furuya Shuichi
Published: March 2004

Instead of conducting trials in absentia, the International Criminal Tribunal for the Former Yugoslavia has an innovative procedure against an accused not in custody. Officially, this ‘Rule 61 procedure’ is only a proceeding for reconfirming an indictment, which does not determine an accused's guilt or innocence. In substance, however, it has functioned as a trial-like procedure to some extent. As a judicial finding on legal issues, the decisions under the procedure have influenced subsequent trials. This article traces the evolution of the procedure and explores its unique character. In particular, by examining essential problems thereof, this article clarifies some points to be considered in establishing a similar mechanism for the International Criminal Court.
The Institution of the International Criminal Court
Authors: Strijards Gerard
Published: March 2004

This article discusses certain key aspects arising from the negotiations leading up to the adoption of a Statute for an International Criminal Court (ICC), to have its seat in The Hague. These aspects include individual criminal responsibility regardless of status as Head of State or constitutional organ and the transformation of international criminal law into domestic law. Also discussed are the two appendices to be added to the Statute pertaining to substantive criminal law and rules of criminal evidence and procedure to be used by the Court. The author argues that the appendix on the law of criminal procedure will be of particular importance to the Netherlands as the host state. The obligations regarding legal assistance of the host state will be dependent on this.
We Did It? The Vertigo of Law and Everyday Life at the Diplomatic Conference on the Establishment of an International Criminal Court
Authors: Tallgren Immi
Published: March 2004

International law is a projection of the imagination and professional identity of the practitioners in the field. Take a diligent, somewhat idealistic Delegate X who participates in a major event in international law: the negotiations on the Statute of the International Criminal Court (ICC), starting with the Ad Hoc Committee in 1995, continuing in the Preparatory Committee and culminating at the Diplomatic Conference in Rome in 1998. What takes place when the ambitious ‘world public opinion’ expectations for international law are supposed to be made everyday life by the delegates? How does X, a single committed person, play his or her role as a tiny part of the global reform machinery? What is the anatomy of the engagement: professionalism and a comfortable exit from the intolerable world of human suffering and injustice? What kind of mechanisms govern the vertiginous scenes of genocide, champagne and flirt? Do the everyday worlds of violence and of welfare touch each other only slightly, in the name of international law, just in order to disappear again in their own, irresistible orbits?
Moving Into UNchartered Waters: An Emerging Right of Unilateral Humanitarian Intervention?
Authors: Blockmans Steven
Published: March 2004

With the black letter law of the UN Charter denying states to unilaterally intervene in third states on humanitarian grounds, this article tries to project a picture of the moral controversy of humanitarian intervention as a balance for order and justice. The author argues that some post-cold war armed interventions may be taken as evidence of an emerging rule of international law outside the UN Charter system allowing the use of unilateral humanitarian intervention to keep a third state from committing large-scale human rights violations on its own territory. However, in the absence of prior authorization from the relevant UN organs, it is necessary to address concerns of possible abuse and manipulations of such an emerging rule. The article includes recommendations to this end.

To go to war for an idea, if the war is aggressive, not defensive, is as criminal as to go to war for territory or revenue; for it is as little justifiable to force our ideas on other people, as to compel them to submit to our will in any other respect. But there assuredly are cases in which it is allowable to go to war, without having been ourselves attacked, or threatened with attack; and it is very important that nations should make up their minds in time, as to what these cases are.John Stuart Mill, A Few Words on Non-Intervention (1859)
Gone with the War? Neutral State Responsibility and the Geneva Arbitration of 1872
Authors: Chadwick Elizabeth
Published: March 2004

The Geneva Arbitration of 1872 was convened to settle various differences between the United States and Great Britain and, in particular, American allegations of British collusion with regard to shipbuilding for the Southern Confederacy during the American Civil War. The Arbitrators ultimately found Britain liable, and awarded $15,500,000 to the United States. This decision remains controversial to the extent that it rested on rules which were not yet accepted as principles of general international law, and which clearly favoured the case of the United States from the outset. It is thus the purpose of this article to explore the facts behind the Geneva Arbitration, and to argue that the finding of British liability in Geneva marked the beginning decline of the laws of neutrality.
Neutral Countries […] may be exploited by the Great Powers both strategically and as a source of additional armies and fleets. Of central importance to the game are those Neutral Countries and provinces which are designated as “Supply Centres.” […] A player's fighting strength is directly related to the number of Supply Centres he or she controls, whilst the game is won when one player controls at least 18 Supply Centres.
Acceptance of International Supervision of Human Rights
Authors: Schermers Henry G.
Published: March 2004

The present article traces the developments with respect to questions of admissibility of claims brought before the European Commission of Human Rights and the European Court of Human Rights by private individuals. It would appear that over the years, the Commission and Court have dealt with an increasing number of cases, and have extended the scope of the infringement they permitted themselves to make on the domestic jurisdiction of states members to the European Convention of Human Rights.
List of Current Proceedings: Update
Authors: Amaya-Castro Juan M.
Published: March 2004

Indonesia and Malaysia jointly seized the Court on 2 November 1998 of their dispute concerning sovereignty over Pulau Ligitan and Pulau Sipadan, two islands in the Celebes Sea. They did so by notifying the Court of a Special Agreement, which was signed between them on 31 May 1997 at Kuala Lumpur and entered into force on 14 May 1998. In the Special Agreement, the Parties request the Court “to determine on the basis of the treaties, agreements and any other evidence furnished by [them], whether sovereignty over Pulau Ligitan and Pulau Sipadan belongs to the Republic of Indonesia or to Malaysia.” They express the wish to settle their dispute “in the spirit of friendly relations existing between [them] as enunciated in the 1976 Treaty of Amity and Co-operation in Southeast Asia” and declare in advance that they will “accept the Judgment of the Court given pursuant to [the] Special Agreement as final and binding upon them.”
Commentary: The Advisory Opinion on the Difference Relating to Immunity from Legal Process of a Special Rapporteur of the Commission of Human Rights: Who Has the Last Word on Judicial Independence?
Authors: Fox Hazel
Published: March 2004

A critical analysis, considering first, the legal competence and propriety of the Court in giving an opinion pursuant to the dispute settlement machinery of the Convention on the Privileges and Immunities of United Nations, whereby advisory jurisdiction over disputes of the UN is equated to contentious jurisdiction between consenting states; and second, the effect in municipal law of a state's obligation to respect the UN Secretary-General's certificate that a UN expert is entitled to immunity from legal process. The Court preserves resort to local courts but requires communication of the UN certificate and immunity to be dealt with expeditiously as a preliminary issue.
List of current legal proceedings: Update
Authors: Pigott Sarah-Jane
Published: March 2004

On 30 August 1999, by leave of the confirming Judge, the Prosecution filed an amended indictment against Dragan Kolundžija.
Constitutional and Institutional Developments
Authors: Rydberg Åsa
Published: March 2004

On 23 July 1999, an Agreement on the enforcement of sentences of the International Criminal Tribunal for the former Yugoslavia (ICTY) was concluded between the United Nations and the Federal Government of Austria. The formalization of the Agreement took place during a meeting in Vienna between Judge Gabrielle Kirk McDonald, President of the ICTY, H.E. Mrs. Benita Ferrero-Waldner, State Secretary, Federal Ministry for Foreign Affairs of Austria and H.E. Mr. Nikolaus Michalek, Federal Minister for Justice of Austria. Austria is the fifth state to enter into such an Agreement.
Commentary: The International War Crimes Trial of Anto Furundžija: Major Progress Toward Ending the Cycle of Impunity for Rape Crimes
Authors: Ashkin Kelly D.
Published: March 2004

This article reviews and analyzes the Furundžija Judgment rendered by the International Criminal Tribunal for the former Yugoslavia, the first international war crimes trial in history to focus virtually exclusively on rape. The judgment addresses what acts constitute rape, whether a non-physical perpetrator can be held individually criminally responsible for rape, and whether rape can constitute torture; it also broadens the purview of Common Article 3 crimes and considers acts which may constitute outrages upon personal dignity. This article discusses how these issues impact upon both international humanitarian law generally and gender jurisprudence specifically.
Mandatory Compliance Powers vis-à-vis States by the Ad Hoc Tribunals and the International Criminal Court: A Comparative Analysis
Authors: Henquet Thomas
Published: March 2004

In the absence of police powers, the International Criminal Court, like the Tribunals for the former Yugoslavia and Rwanda, will depend on the co-operation of states in order to fulfil its mandate. Discussing the jurisprudence of the ad hoc Tribunals and the travaux préparatoires of the Rome treaty, the author compares the mandatory powers conferred on the respective institutions to this end. He concludes that the Security Council endowed the Tribunals with unequivocally binding powers, while under the Rome treaty regime, which resembles traditional inter-state co-operation in criminal matters, the ICC's powers are more limited and state obligations less stringent.
Dealing With Crimes of a Past Regime. Is Amnesty Still an Option?
Authors: Dugard John
Published: March 2004

From time immemorial amnesty has been employed as a means of promoting a political settlement and advancing reconciliation in societies that have emerged from repression. At present there is a trend in support of prosecution of those who have committed international crimes, such as torture and crimes against humanity, which excludes the possibility of amnesty. That amnesty is no longer favored is illustrated by the failure of the Rome Statute of the International Criminal Court to recognize amnesty as a defence to prosecution. While there is no place for unconditional amnesty in the contemporary international legal order an intermediate solution such as a Truth and Reconciliation Commission with power to grant amnesty after investigation, of the South African kind, may contribute to the achievement of peace and justice in a society in transition more effectively than mandatory prosecution.
Piacentini's Window: The Modernism of the Fascist Master Plan of Rome
Authors: BAXA PAUL
Published: March 2004

When urban planners Marcello Piacentini and Antonio Muñoz looked at the Rome they helped create through their participation in the Master Plan of 1931, they saw a metaphor for Fascism's transformation of Rome, a metaphor that captured the ‘effect’ of the Master Plan but not its ‘intention’. The Fascist regime aimed to build a modern capital on the nineteenth-century models of Paris and Vienna, but created instead a modernist city which challenged, on many levels, the neo-classicist rationalism of the previous century's urban planning. This article explores the reasons for such a disjuncture between intention and effect and finds them within Fascist ideology and experience.
Germans in the Eyes of the Gestapo: The Ciechanów District, 1939–1945
Authors: GRABOWSKI JAN , GRABOWSKI ZBIGNIEW R.
Published: March 2004

The files of the Ciechanów (Zichenau) Gestapo – one of the few remaining archives of this kind from German-occupied Poland – offer interesting insights into the social policy of the Nazi state. The Germanisation of Polish territories occurred by deporting and exterminating the Jews, depriving Poles of their rights and supporting the local Germans and the ethnic Germans resettled from the East. The German minority living in this ethnically mixed region was required to adhere to strict codes of behaviour and was held accountable for all unauthorised contacts with their Polish and, even more so, their Jewish neighbours. The system of control and repression strove to isolate the various ethnic (‘racial’) groups, encouraging denunciations and thus instilling fear in the populace. This article pays particular attention to the actions of German citizens who fell under the scrutiny of the Secret Police.
For a Republic ‘Diverse and Indivisible’? France's Experience from the Colonial Past
Authors: DIMIER VÉRONIQUE
Published: March 2004

The constitutional debate around the establishment of the Union Française (1946) is examined here as part of a long-term debate between two competing definitions of the nation and the republic: an integrating and ‘jacobin’ one, which aimed at moulding the colonial empire along the lines of the ‘one and indivisible’ republic, and another, ‘Girondin’ and contextual one, that saw the Union Française as a possible means of carving out a diverse, possibly divisible, republic. Two different ways of considering both French citizenship and the hybrid citizenship of the Union Française as put together in the Constitution of 1946 developed out of those two conceptions.
The Rise and Fall of Western Europe's Democratic Age, 1945–1973
Authors: CONWAY MARTIN
Published: March 2004

Why did western Europe become so suddenly democratic after 1945? After the upheavals of the previous decade the rather placid politics that follows the war is at first sight difficult to explain. This article seeks to go beyond the tendency of much historical writing to see the hegemonic parliamentary democracy of the roughly twenty-five years after 1945 as the product of exhaustion, economic prosperity or the constraints imposed by the Cold War. Instead, it argues that a path towards democracy can be detected within the events of the war years which then came to fruition in the rather conservative and limited democratic structures of the postwar decades. This Democratic Age then came to a conclusion in the renewed contestation of the late 1960s and early 1970s.
Reply to Jones
Authors: CONWAY MARTIN
Published: March 2004

Historians are from Mars and political scientists are from Venus (or was it the other way round?). The most striking feature of Erik Jones's stimulating and generous response to my article is the way in which it highlights the divergent approaches to the postwar era adopted by historians and political scientists. In many respects, this is a very good thing. We need the stimuli provided by those rooted in different traditions bringing their distinctive approaches to the same subject matter, just as, for the early medieval period, historians, archaeologists and literary scholars confront the fragmentary evidence of post-Roman Europe in contrasting but often mutually enriching ways.
The Transformation of European Universities: Disciplines and Professions in England, Germany and Russia since 1870
Authors: WITTROCK BJÖRN
Published: March 2004

Dietrich Beyrau, ed., Im Dschungel der Macht. Intellektuelle Professionen unter Stalin und Hitler (Göttingen: Vandenhoeck & Ruprecht, 2000), 399 pp., €52.00 (hb.), ISBN 3-525-36244-7.

Corine Defrance, Les alliés occidentaux et les universités allemandes, 1945–1949 (Paris: CNRS Éditions, 2000), 406 pp., €27.50, ISBN 2-271-05768-X.

Ralph Jessen, Akademische Elite und kommunistische Diktatur. Die ostdeutsche Hochschullehrerschaft in der Ulbricht-Ära (Göttingen: Vandenhoeck & Ruprecht, 1999), 552 pp., €52.00, ISBN 3-525-35797-4.

Brian Pullan with Michele Abendstern, A History of the University of Manchester, 1951–73 (Manchester: Manchester University Press, 2000), 281 pp., €32.95, ISBN 0-7190-5670-5.

Reba N. Soffer, Discipline and Power: The University and the Making of an English Elite, 1870–1930 (Stanford: Stanford University Press, 1994), 308 pp., €55.00, ISBN 0-8047-2383-4.
Ionic calcium levels in oviduct explant-conditioned media from an Australian marsupial, the brushtail possum (Trichosurus vulpecula) and its relevance to in vitro fertilization
Authors: Sidhu K.S. , Mate K.E. , Molinia F.C. , Berg D.K. , Rodger J.C.
Published: March 2004

Gametes from the brushtail possum (Trichosurus vulpecula), an Australian marsupial, require exposure to oviductal cells and/or their secretions before sperm binding and penetration of the zona pellucida can occur. Sperm-egg fusion, the next critical step in fertilization has not previously been reported in vitro. Here we describe the refinement of an oviduct epithelial cell (OEC) explant culture system using two different media to obtain in vitro sperm-egg fusion in the brushtail possum for the first time. Conditioned media from OEC explant cultures were supplemented with either 1% fetal calf serum (FCS) or 1mg/ml polyvinyl alcohol and used for co-culture of epididymal sperm and superovulated eggs. Under these conditions zona penetration rates varied from 0 to 46% and sperm-egg fusion from 0 to 20%. Analysis of explant conditioned media indicated that qualitative and quantitative differences between batches could account, at least partially, for the large variability in zona penetration rates. Conditioned media that contained approximately 1mM of ionic calcium were most effective for achieving sperm capacitation, zona binding, and penetration and sperm-egg fusion. The reorientation of the sperm head to T-shape, an indicator of capacitation in the brushtail possum, was closely linked with the concentration of calcium present in vitro.
Human oocytes and preimplantation embryos express mRNA for growth hormone receptor
Authors: Ménézo Y.J.R. , El Mouatassim S. , Chavrier M. , Servy E.J. , Nicolet B.
Published: March 2004

Human genetic expression of growth hormone receptor (GHR) gene was qualitatively analysed using reverse transcription polymerase chain reaction (RT-PCR) in cumulus cells, immature germinal vesicle (GV) and mature metaphase II (MII) stage oocytes and preimplantation human embryos. The transcripts encoding GHR were detected in cumulus cells and also in naked oocytes, either mature or not. In this case, a nested PCR is needed, as for early embryo preimplantation stages, before genomic activation. The GHR gene is highly expressed from the 4-day morula onwards. This suggests that GHR transcription follows a classical scheme associated with genomic activation. It is probable that, in human, growth hormone plays a role in the final stages of oocyte maturation and early embryogenesis as it does for several other mammalian species.
Maturation of porcine oocytes after cooling at the germinal vesicle stage
Authors: Liu Rui-Hua , Sun Qing-Yuan , Li Yong-Hai , Jiao Li-Hong , Wang Wei-Hua
Published: March 2004

Maturation of porcine oocytes was examined after oocytes were cooled at the germinal vesicle stage. Cumulus-oocyte complexes (COCs) collected from medium-sized follicles were cooled at 24 °C or 4 °C for 5, 30 or 120 min in a solution with or without 1.5 M dimethylsulfoxide (DMSO). After rewarming, COCs were cultured in maturation medium at 39 °C, 5% CO2 in air for 44 h. Meiotic spindle organisation (by immunostaining and confocal microscopy), nuclear maturation (by orcein staining) and cytoplasmic maturation (by intracellular glutathione assay) of oocytes were examined after maturation. When COCs were cooled at 24 °C for various times in the medium without DMSO, a tendency to decreased spindle formation, nuclear maturation and cytoplasmic maturation was observed, but there was no statistical difference compared with controls. Addition of DMSO during cooling inhibited subsequent nuclear maturation and spindle formation. When COCs were cooled at 4 °C, both nuclear and cytoplasmic maturation as well as spindle formation were inhibited in most oocytes in a time-dependent manner. DMSO during cooling did not have any beneficial effect on subsequent oocyte maturation and spindle formation. These results suggest that porcine oocytes are very sensitive to a drop in the temperature before exposure to culture. Cooling oocytes before maturation inhibits their subsequent spindle organisation, nuclear and cytoplasmic maturation. Addition of DMSO to the cooling solution did not protect porcine oocytes from cooling-induced damage.
The fertilising ability of spermatogenic cells derived from cultured mouse immature testicular tissue
Authors: Suzuki Sayuri , Sato Kahei
Published: March 2004

There are many reports about the in vitro culture of spermatogenic cells, but no-one has succeeded in inducing the differentiation from spermatogonia to intact sperm. Also the study of in vitro testicular tissue culture has hardly advanced. We studied the culture of mouse immature testicular tissue derived from 5-day-old mice. We aimed to achieve the differentiation of spermatogenic cells in order to observe spermatogenesis in testicular tissue in vitro. We also froze mature testicular tissue and immature testicular tissue cultured for 2 weeks. Furthermore, spermatogenic cells differentiated by culturing were injected into metaphase II oocytes to determine whether these differentiated cells and frozen-thawed testicular tissue have fertilising and developmental ability. Under the culture conditions employed, secondary spermatocytes and a few round spermatids differentiated from spermatogonia were observed in the immature testicular tissue cultured for 2 weeks. When spermatogenic cells derived from cultured immature testicular tissue, cultured frozen immature testicular tissue and frozen-thawed mature testicular tissue were injected into ooplasm, the oocytes were fertilised and fertilised oocytes developed to the 8-cell stage. We suggest that spermatogenic cells derived from cultured immature testicular tissue have fertilising and developmental abilities equivalent to that of sperm. Also these abilities of spermatogenic cells obtained from cultured frozen immature testicular tissue and frozen-thawed mature testicular tissue were better than those of the same cells before freezing.
Birth of rats following nuclear exchange at the 2-cell stage
Authors: Roh Sangho , Guo Jitong , Malakooti Nakisa , Morrison John R. , Trounson Alan O. , Du Zhong Tao
Published: March 2004

We report full-term development of nuclear transfer embryos following nuclear exchange at the 2-cell stage. Nuclei from 2-cell rat embryos were transferred into enucleated 2-cell embryos and developed to term after transfer to recipients (NT2). Pronuclear exchange in zygotes was used for comparison (NT1). Zygotes and 2-cell embryos were harvested from 4-week-old female Sprague-Dawley rats. Nuclear transfer was performed by transferring the pronuclei or karyoplasts into the perivitelline space of recipient embryos followed by electrofusion to reconstruct embryos. Fused couplets were cultured for 4 or 24 h before being transferred into day 1 pseudopregnant recipients (Hooded Wistar) at the 1- or 2-cell stage. In vitro culture was also carried out to check the developmental competence of the embryos. In vitro development to the blastocyst stage was not significantly different between the two groups (NT1, 34.3%; NT2, 45.0%). Two of three recipients from NT1 and two of five recipients from NT2 became pregnant. Six pups (3 from NT1, 3 from NT2) were delivered from the four foster mothers. Three female pups survived; 2 from NT1 and 1 from NT2. At 2 months of age these pups appeared healthy, and were mated with Sprague-Dawley males. One rat derived from NT1 delivered 15 pups (5 males, 10 females) as did the rat from NT2 (7 males, 8 females). Our results show that by using karyoplasts from 2-cell stage embryos as nuclear donors and reconstructing them with enucleated 2-cell embryos, healthy rats can be produced.
HDAC6-induced premature chromatin compaction in mouse oocytes and fertilised eggs
Authors: Verdel André , Seigneurin-Berny Daphné , Faure Anne-Karen , Eddahbi Mina , Khochbin Saadi , Nonchev Stefan
Published: March 2004

Chromatin remodelling in the fertilised mouse egg is intimately linked to protein synthesis and degradation, to protamine by histone replacement and to specific histone modifications. The involvement of histone deacetylases (HDACs) in the beginning of development is poorly understood. HDACs are essential for cell proliferation and in the control of gene expression in a wide variety of mammalian systems. Here we focus on mHDAC6, a recently identified class II histone deacetylase, and we analyse its expression and localisation in oocytes and pronuclear zygotes. By indirect immunofluorescence we show that mHDAC6 is detected in the cytoplasm of germinal vesicle (GV) stage oocytes and 1-cell embryos. Ectopic expression of this enzyme after injection into germinal vesicles and pronuclei alters the nuclear structure and causes premature compaction of the chromatin. Our data suggest that the effect of condensation is linked to the ubiquitin-binding activity of mHDAC6, rather than to its function as a deacetylase.
Effects of PKC activation on the meiotic maturation, fertilization and early embryonic development of mouse oocytes
Authors: Quan He-Mei , Fan Heng-Yu , Meng Xiao-Qian , Huo Li-Jun , Chen Da-Yuan , Schatten Heide , Yang Pei-Man , Sun Qing-Yuan
Published: March 2004

Protein kinase C (PKC) is a family of Ser/Thr protein kinase widely distributed in eukaryotes. There is evidence that PKC plays key roles in the meiotic maturation and activation of mammalian oocytes. However, the mechanism of PKC's actions and the PKC isoforms responsible for these actions are poorly understood. In this study, we reveal in mouse eggs and early embryos: (1) the effects of PKC on the meiotic and mitotic cell cycle progression during oocyte maturation, egg activation and embryonic cleavages; (2) the functional importance of classical PKC subclasses in these processes; and (3) the subcellular localization of the PKCα isoform during development from GV stage oocytes to the blastocyst stage embryos. The results indicate that the PKC activator phorbol 12-myristate 13-acetate (PMA) inhibits the meiotic resumption of cumulus-free mouse oocytes by a mechanism dependent not only on classical PKC activity but also on other PKC isoforms. PKC activation after germinal vesicle breakdown leads to the inhibition of mitogen-activated protein kinase phosphorylation and the arrest of cell cycle at MI stage. The second polar body emission and the cleavages of early embryos are blocked after prolonged PKC activation. The subcellular localization of PKCα isoform in mouse oocytes and embryos is developmental-stage associated. All these results suggest that PKC has multiple functional roles in the cell cycle progression of mouse oocytes and embryos.
Supplementation with cysteamine during maturation and embryo culture on embryo development of prepubertal goat oocytes selected by the brilliant cresyl blue test
Authors: Urdaneta Aixa , Jiménez-Macedo Ana-Raquel , Izquierdo Dolors , Paramio Maria-Teresa
Published: March 2004

Our previous studies have shown that the addition of 100 μM cysteamine to the in vitro maturation (IVM) medium increased the embryo development of prepubertal goat oocytes. The aim of the present study was to evaluate the effect of adding different concentrations of cysteamine to the IVM medium and to the in vitro embryo culture medium (IVC) on the embryo development of prepubertal goat oocytes selected by the brilliant cresyl blue (BCB) test. Oocytes were exposed to BCB and classified as: oocytes with a blue cytoplasm or grown oocytes (BCB+) or oocytes without blue cytoplasm or growing oocytes (BCB−). In Experiment 1, oocytes were matured in a conventional IVM medium supplemented with 100 μM, 200 μM or 400 μM cysteamine. In Experiment 2, oocytes were matured with 400 μM cysteamine and following in vitro fertilization (IVF) were cultured in SOF medium supplemented with 50 μM and 100 μM cysteamine. In Experiment 1, BCB+ oocytes matured with 100 μM and 200 μM cysteamine showed higher normal fertilization and embryo development rates than BCB− oocytes. Oocytes matured with 400 μM cysteamine did not present these differences between BCB+ and BCB− oocytes. In Experiment 2, the addition of 50 μM and 100 μM cysteamine to culture medium did not affect the proportion of total embryos obtained from BCB+ oocytes (35.89% and 38.29%, respectively) but was significantly different in BCB− oocytes (34.23% and 29.04%, respectively, P<0.05). In conclusion, the addition of 400 μM cysteamine to the IVM improved normal fertilization and embryo development of BCB− oocytes at the same rates as those obtained from BCB+ oocytes. The proportions of morulae plus blastocyst development were not affected by the treatments.
Survival of rapidly frozen hatched mouse blastocysts
Authors: Pribenszky Csaba , Cseh Sándor , Abonyi-Tóth Zsolt , Solti László
Published: March 2004

The objective of the present study was to examine the effect of rapid freezing on the in vitro and in vivo survival of zona-pellucida-free hatched mouse blastocysts. Hatched blastocysts were rapidly frozen in a freezing medium containing either ethylene glycol (EG) or glycerol (G) in 1.5 M or 3 M concentration. Prior to freezing, embryos were equilibrated in the freezing medium for 2 min, 10 min, 20 min or 30 min at room temperature. To freeze them, embryos were held in liquid nitrogen vapour [≈1 cm above the surface of the liquid nitrogen (LN2)] for 2 minutes and then immersed into LN2. After thawing, embryos were transferred either to rehydration medium (DPBS + 10% foetal calf serum + 0.5 M sucrose) for 10 minutes or rehydrated directly in DPBS supplemented with foetal calf serum. In vitro survival of embryos frozen with EG was higher than those frozen with G. The highest survival was obtained with 3 M EG and 2 min or 10 min equilibration prior to freezing, combined with direct rehydration after thawing. Frozen blastocysts developed into normal foetuses as well as unfrozen control ones did, with averages of 30% (control), 26% (EG) and 15% (G). The results show that hatching and hatched mouse blastocysts can be cryopreserved by a simple rapid freezing protocol in EG without significant loss of viability. Our data indicate that the mechanical protection of the zona pellucida is not needed during freezing in these stages.
Fertilization following intracytoplasmic sperm injection of in vivo and in vitro matured oocytes from an Australian marsupial, the tammar wallaby (Macropus eugenii)
Authors: Magarey Genevieve M. , Mate Karen E.
Published: March 2004

Conventional in vitro fertilization (IVF) techniques have been unable to produce normal embryos in any Australian marsupial, largely owing to problems with the early stages of sperm–oocyte binding. This study has used intracytoplasmic sperm injection (ICSI) of in vivo and in vitro matured tammar wallaby oocytes to bypass these processes and achieve fertilization in vitro. The fertilization rate (i.e. development to the two-pronuclei stage) of in vivo and in vitro matured oocytes following ICSI and sham injection was assessed at 17–19 h after injection. Fertilization occurred in 48% (45/93) of in vivo matured oocytes that were injected with spermatozoa. Significantly fewer sham-injected oocytes (6/82, P<0.005) and uninjected control oocytes (5/84, P<0.005) formed two pronuclei. In a direct comparison, the numbers of in vivo and in vitro matured oocytes that formed two pronuclei after ICSI were 22/28 (78.6%) and 23/40 (57.6%), respectively, which are not significantly different. There was also no significant difference in the nuclear response of in vivo and in vitro matured oocytes to sham injection. The numbers of oocytes forming a single pronucleus after sham injection were 10/24 (41.7%) and 24/37 (64.9) for in vivo and in vitro matured oocytes, respectively. Immature germinal-vesicle-stage oocytes were unable to decondense sperm injected during ICSI or to form pronuclei. These results demonstrate that both in vitro and in vivo matured tammar wallaby oocytes can be fertilized by ICSI. The success of ICSI not only offers the opportunity for fundamental analysis of marsupial fertilization but could, in conjunction with development of appropriate culture conditions and embryo transfer technologies, contribute to increased production of offspring from rare or valuable marsupials.
Expression of early development-related genes in bovine nuclear transferred and fertilized embryos
Authors: Park Sang Hyun , Park Soo-Bong , Kim Nam-Hyung
Published: March 2004

Cloning efficiency following somatic cell nuclear transfer is very low. In order to obtain insights into this problem, mRNA expression patterns of early development-related genes in nuclear transferred embryos were compared with those obtained from in vivo and in vitro fertilization. Semiquantitative reverse-transcription polymerase chain reaction assay was used to compare the gene expression of, the cell adhesion protein E-cadherin, interleukin -6, heat-shock protein 70.1 and bos taurus apoptosis regulator box-a (Bax). The relative abundances of glucose transporter-1, E-cadherin and interleukin-6 were significantly (P<0.05) higher in in vitro fertilized morulae than in vivo derived morulae. Transcription of the gene encoding octamer-binding transcription factor 4 was higher in blastocysts obtained from in vivo fertilization than in those from in vivo blastocysts. The transcript for Bax was markedly upregulated in blastocysts derived from in vitro production and nuclear transfer procedures compared with in vivo fertilization. These results suggest that alterations in mRNA expression of early development genes are more associated with in vitro culture condition than the nuclear transfer procedure itself.
Primary pulmonary hypertension: molecular basis and potential for therapy
Authors: Rudarakanchana Nung , Morrell Nicholas W.
Published: March 2004

Primary pulmonary hypertension (PPH) is defined clinically by sustained elevation of pulmonary arterial pressure without a demonstrable cause, and is a progressive, often-fatal disease. PPH can be associated with ingestion of appetite suppressants, human immunodeficiency virus infection and certain autoimmune diseases. Familial PPH is known to account for 6% of all cases. Mutations in the gene encoding the bone morphogenetic protein (BMP) type II receptor have been identified in 72% of affected families and 26% of apparently sporadic cases. BMPs are members of the transforming growth factor β superfamily and affect intracellular signalling via Smads and mitogen-activated protein kinases. Evidence supports a ‘two-hit’ hypothesis in which PPH is triggered by accumulation of genetic and environmental insults in a susceptible individual. Elucidation of the precise molecular and cellular mechanisms underlying PPH will provide a powerful basis for the development of novel therapeutic strategies in the treatment of this devastating condition.
A spectroscopy and isotope study of sediments from the Antarctic Dry Valleys as analogues for potential paleolakes on Mars
Authors: Bishop Janice L. , Anglen Brandy L. , Pratt Lisa M. , Edwards Howell G. M. , Des Marais David J. , Doran Peter T.
Published: March 2004

A spectroscopy and isotope study has been performed on igneous sediments from Lake Hoare, a nearly isolated ecosystem in the Dry Valleys region of Antarctica. The mineralogy and chemistry of these sediments were studied in order to gain insights into the biogeochemical processes occurring in a permanently ice-covered lake and to assist in characterizing potential habitats for life in paleolakes on Mars. Obtaining visible/near-infrared, mid-infrared and Raman spectra of such sediments provides the ground truth needed for using reflectance, emittance and Raman spectroscopy for exploration of geology, and perhaps biology, on Mars. Samples measured in this study include a sediment from the ice surface, lake bottom sediment cores from oxic and anoxic zones of the lake and the magnetic fractions of two samples.

These sediments are dominated by quartz, pyroxene, plagioclase and K-feldspar, but also contain calcite, organics, clays, sulphides and iron oxides/hydroxides that resulted from chemical and biological alteration processes. Chlorophyll-like bands are observed in the spectra of the sediment-mat layers on the surface of the lake bottom, especially in the deep anoxic region. Layers of high calcite concentration in the oxic sediments and layers of high pyrite concentration in the anoxic sediments are indicators of periods of active biogeochemical processing in the lake. Micro-Raman spectra revealed the presence of ~5 μm-sized pyrite deposits on the surface of quartz grains in the anoxic sediments. C, N and S isotope trends are compared with the chemistry and spectral properties. The δ13C and δ15N trends highlight the differences in the balance of microbial processes in the anoxic sediments versus the oxic sediments. The biogenic pyrite found in the sediments from the anoxic zone is associated with depleted δ34S values, high organic C levels and chlorophyll spectral bands and could be used as a potential biomarker mineral for paleolakes on Mars.
The sulfocyanic theory on the origin of life: towards a critical reappraisal of an autotrophic theory
Authors: Perezgasga L. , Silva E. , Lazcano A. , Negrón-Mendoza A.
Published: March 2004

In the early 1930s, Alfonso L. Herrera proposed his so-called sulfocyanic theory on the origin of life, an autotrophic proposal on the first living beings according to which NH4SCN and H2CO acted as raw materials for the synthesis of bio-organic compounds inside primordial photosynthetic protoplasmic structures. Although the work of Herrera is frequently cited in historical analysis of the development of the origin of life studies, very little attention has been given to the chemical significance of the reactions he published. In this paper we report the results of our search for amino acids obtained from a reactive mixture used by Herrera from 1933 onwards. Chromatograms using the high-pressure liquid chromatography (HPLC) technique suggest the presence of several amino acids, the total yield being 2% of the initial thiocyanate used. Preliminary identification based on HPLC retention times suggests the presence of glycine, alanine, cysteine and methionine. Alanine was the most abundant amino acid in all samples of fractionated material analysed. Although the starting materials used by Herrera were determined by his autotrophic hypothesis on the origin of cells, our results show that his experiments may provide insights into the abiotic synthesis of sulfur-containing amino acids within the framework of a heterotrophic emergence of life.
Testing evolutionary convergence on Europa
Authors: Chela-Flores J.
Published: March 2004

A major objective in solar system exploration is the insertion of appropriate biology-oriented experiments in future missions. We discuss various reasons for suggesting that this type of research be considered a high priority for feasibility studies and, subsequently, for technological development of appropriate melters and submersibles. Based on numerous examples, we argue in favour of the assumption that Darwin's theory is valid for the evolution of life anywhere in the universe. We have suggested how to obtain preliminary insights into the question of the distribution of life in the universe. Universal evolution of intelligent behaviour is at the end of an evolutionary pathway, in which evolution of ion channels in the membrane of microorganisms occurs in its early stages. Further, we have argued that a preliminary test of this conjecture is feasible with experiments on the Europan surface or ocean, involving evolutionary biosignatures (ion channels). This aspect of the exploration for life in the solar system should be viewed as a complement to the astronomical approach for the search of evidence of the later stages of the evolutionary pathways towards intelligent behaviour.
The evolution of habitable zones during stellar lifetimes and its implications on the search for extraterrestrial life
Authors: Underwood D.R. , Jones B.W. , Sleep P.N.
Published: March 2004

A stellar evolution computer model has been used to determine changes in the luminosity L and effective temperature Te of single stars during their time on the main sequence. The range of stellar masses investigated was from 0.5 to 1.5 times that of the Sun, each with a mass fraction of metals (metallicity, Z) from 0.008 to 0.05. The extent of each star's habitable zone (HZ) has been determined from its values of L and Te. These stars form a reference framework for other main sequence stars. All of the 104 main sequence stars known to have one or more giant planets have been matched to their nearest stellar counterpart in the framework, in terms of mass and metallicity, hence closely approximating their HZ limits. The limits of HZ, for each of these stars, have been compared to their giant planet(s)'s range of strong gravitational influence. This allows a quick assessment as to whether Earth-mass planets could exist in stable orbits within the HZ of such systems, both presently and at any time during the star's main sequence lifetime. A determination can also be made as to the possible existence of life-bearing satellites of giant planets, which orbit within HZs. Results show that about half of the 104 known extrasolar planetary systems could possibly have been housing an Earth-mass planet in HZs during at least the past billion years, and about three-quarters of the 104 could do so for at least a billion years at some time during their main sequence lives. Whether such Earth-mass planets could have formed is an urgent question now being investigated by others, with encouraging results.